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Sample records for pretreatment ebv dna

  1. The clinical significance of EBV DNA in the plasma and peripheral blood mononuclear cells of patients with or without EBV diseases.

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    Kanakry, Jennifer A; Hegde, Aparna M; Durand, Christine M; Massie, Allan B; Greer, Amy E; Ambinder, Richard F; Valsamakis, Alexandra

    2016-04-21

    Epstein-Barr virus (EBV) is a ubiquitous virus that establishes a latent infection within the host and in some cases can lead to the development of EBV-associated lymphomas, lymphoproliferative disorders, hemophagocytic lymphohistiocytosis, solid tumors, and other diseases. We studied the clinical significance of detecting EBV DNA in the plasma and peripheral blood mononuclear cells (PBMCs) of 2146 patients who had blood specimens sent to the Johns Hopkins Hospital clinical laboratory for viral quantitative real-time polymerase chain reaction assay over a 5-year period. Within this largely immunocompromised and hospitalized cohort, 535 patients (25%) had EBV detected in plasma or PBMCs. When EBV was detected in the absence of an EBV(+)disease (n = 402), it was present only in PBMCs in 69% of cases. Immunocompromised patients were less likely to have EBV in plasma than in PBMCs in the absence of EBV(+)disease. In patients with active, systemic EBV(+)diseases (n = 105), EBV was detected in plasma in 99% of cases but detected in PBMCs in only 54%. Across a range of copy number cutoffs, EBV in plasma had higher specificity and sensitivity for EBV(+)disease as compared with EBV in PBMCs. EBV copy number in plasma distinguished untreated, EBV(+)lymphoma from EBV(+)lymphoma in remission and EBV(-)lymphoma, and also distinguished untreated, EBV(+)posttransplantation lymphoproliferative disorder (PTLD) from EBV(+)PTLD in remission and EBV(-)PTLD. EBV copy number quantification is a useful diagnostic marker across the spectrum of EBV(+)diseases, even among immunocompromised patients, with plasma specimens more indicative of EBV(+)disease than PBMCs. © 2016 by The American Society of Hematology.

  2. Variable EBV DNA Load Distributions and Heterogeneous EBV mRNA Expression Patterns in the Circulation of Solid Organ versus Stem Cell Transplant Recipients

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    A. E. Greijer

    2012-01-01

    Full Text Available Epstein-Barr virus (EBV driven post-transplant lymphoproliferative disease (PTLD is a heterogeneous and potentially life-threatening condition. Early identification of aberrant EBV activity may prevent progression to B-cell lymphoma. We measured EBV DNA load and RNA profiles in plasma and cellular blood compartments of stem cell transplant (SCT; n=5, solid organ transplant recipients (SOT; n=15, and SOT having chronic elevated EBV-DNA load (n=12. In SCT, EBV DNA was heterogeneously distributed, either in plasma or leukocytes or both. In SOT, EBV DNA load was always cell associated, predominantly in B cells, but occasionally in T cells (CD4 and CD8 or monocytes. All SCT with cell-associated EBV DNA showed BARTs and EBNA1 expression, while LMP1 and LMP2 mRNA was found in 1 and 3 cases, respectively. In SOT, expression of BARTs was detected in all leukocyte samples. LMP2 and EBNA1 mRNA was found in 5/15 and 2/15, respectively, but LMP1 mRNA in only 1, coinciding with severe PTLD and high EBV DNA. Conclusion: EBV DNA is differently distributed between white cells and plasma in SOT versus SCT. EBV RNA profiling in blood is feasible and may have added value for understanding pathogenic virus activity in patients with elevated EBV-DNA.

  3. Variable EBV DNA Load Distributions and Heterogeneous EBV mRNA Expression Patterns in the Circulation of Solid Organ versus Stem Cell Transplant Recipients

    NARCIS (Netherlands)

    Greijer, A. E.; Stevens, S. J.; Verkuijlen, S. A.; Juwana, H.; Fleig, S. C.; Verschuuren, E. A.; Hepkema, B. G.; Cornelissen, J. J.; Brooimans, R. A.; Verdonck, L. F.; Middeldorp, J. M.

    2012-01-01

    Epstein-Barr virus (EBV) driven post-transplant lymphoproliferative disease (PTLD) is a heterogeneous and potentially life-threatening condition. Early identification of aberrant EBV activity may prevent progression to B-cell lymphoma. We measured EBV DNA load and RNA profiles in plasma and cellular

  4. DNA Epstein-Barr virus (EBV sebagai biomaker diagnosis karsinoma nasofaring

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    Janti Sudiono

    2013-09-01

    Full Text Available Background: Nasopharyngeal carcinoma (NPC is a malignant neoplasm arising from the mucosal epithelium of the nasopharynx with various cells differentiation. Nasopharyngeal carcinoma is vastly more common in certain regions of East Asia, South Asia and Africa with viral, dietary which is typically includes consumption of salted vegetables, fish, meat and genetic factors that implicated in its causation. The undifferentiated is the most common type of NPC and strongly associated with Epstein-Barr virus (EBV infection. Purpose: This paper was aimed to review about molecular biomarker as non invasive diagnosis of NPC especially in related to EBV infection in nasopharyngeal epithelial cells. Reviews: The pathogenesis of NPC particularly the endemic type seems to follow a multi-step process, in which EBV, ethnic background, and environmental carcinogens all seem to play important role. EBV DNA plasm level is used continuously in clinic as a promise, sensitive and specific molecular marker diagnostic that reflected the stage, treatment response and prognosis of NPC. Detection of nuclear antigen associated with Epstein-Barr virus (EBNA and viral DNA has revealed that EBV can infect epithelial cells and associated with their transformation in carcinogenesis. Latent membrane protein (LMP-1 and LMP-2 oncogenes EBV encoded related to proliferative gene expression indicated invasive and progressive growth of NPC. Conclusion: The new biomarkers for NPC, including EBV DNA in serum; EBV DNA and BamH1-A Reading Frame-1 (BARF1 mRNA in NPC brushings have been developed for the molecular non invasive diagnosis of this tumour.Latar belakang: Nasopharyngeal carcinoma (NPC, sering dikenal sebagai kanker nasofaring merupakan tumor ganas yang berasal dari epitel mukosa nasofaring dengan derajat diferensiasi sel yang bervariasi. Paling banyak ditemukan di Asia Selatan, Asia Timur, dan Afrika. Virus, pola diet tipikal seperti konsumsi sayuran, ikan dan daging yang

  5. Complex forms of mitochondrial DNA in human B cells transformed by Epstein-Barr virus (EBV)

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    Christiansen, Gunna; Christiansen, C; Zeuthen, J

    1983-01-01

    Human lymphocytes and lymphoid cell lines were analyzed for the presence of complex forms of mitochondrial DNA (mtDNA) by electron microscopy. A high frequency (9%-14.5%) of catenated dimers, circular dimers, or oligomers were found in samples from Epstein-Barr-virus-(EBV) transformed lymphoblast......Human lymphocytes and lymphoid cell lines were analyzed for the presence of complex forms of mitochondrial DNA (mtDNA) by electron microscopy. A high frequency (9%-14.5%) of catenated dimers, circular dimers, or oligomers were found in samples from Epstein-Barr-virus-(EBV) transformed...

  6. Comparison of EBV DNA viral load in whole blood, plasma, B-cells and B-cell culture supernatant.

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    Ouedraogo, David Eric; Bollore, Karine; Viljoen, Johannes; Foulongne, Vincent; Reynes, Jacques; Cartron, Guillaume; Vendrell, Jean-Pierre; Van de Perre, Philippe; Tuaillon, Edouard

    2014-05-01

    Epstein-Barr virus (EBV) genome quantitation in whole blood is used widely for therapeutic monitoring of EBV-associated disorders in immunosuppressed individuals and in patients with EBV-associated lymphoma. However, the most appropriate biological material to be used for EBV DNA quantitation remains a subject of debate. This study compare the detection rate and levels of EBV DNA from whole blood, plasma, enriched B-cells, and B-cell short-term culture supernatant using quantitative real-time PCR. Samples were collected from 33 subjects with either HIV infection or B-cell lymphoma. Overall, EBV DNA was detected in 100% of enriched B-cell samples, in 82% of B-cell culture supernatants, in 57% of plasma, and 42% of whole blood samples. A significant correlation for EBV viral load was found between enriched B-cell and B-cell culture supernatant material (ρ = 0.92; P < 0.0001), but no significant correlation existed between EBV DNA levels in whole blood and enriched B-cells (ρ = -0.02; P = 0.89), whole blood and plasma (ρ = 0.24; P = 0.24), or enriched B-cells and plasma (ρ = 0.08; P = 0.77). Testing of enriched B-cells appeared to be the most sensitive method for detection of EBV DNA as well as for exploration of the cellular reservoir. Quantitation of EBV DNA in plasma and B-cell culture supernatant may be of interest to assess EBV reactivation dynamics and response to treatment as well as to decipher EBV host-pathogen interactions in various clinical scenarios. © 2013 Wiley Periodicals, Inc.

  7. Performance of the Real-Q EBV Quantification Kit for Epstein-Barr Virus DNA Quantification in Whole Blood.

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    Huh, Hee Jae; Park, Jong Eun; Kim, Ji Youn; Yun, Sun Ae; Lee, Myoung Keun; Lee, Nam Yong; Kim, Jong Won; Ki, Chang Seok

    2017-03-01

    There has been increasing interest in standardized and quantitative Epstein-Barr virus (EBV) DNA testing for the management of EBV disease. We evaluated the performance of the Real-Q EBV Quantification Kit (BioSewoom, Korea) in whole blood (WB). Nucleic acid extraction and real-time PCR were performed by using the MagNA Pure 96 (Roche Diagnostics, Germany) and 7500 Fast real-time PCR system (Applied Biosystems, USA), respectively. Assay sensitivity, linearity, and conversion factor were determined by using the World Health Organization international standard diluted in EBV-negative WB. We used 81 WB clinical specimens to compare performance of the Real-Q EBV Quantification Kit and artus EBV RG PCR Kit (Qiagen, Germany). The limit of detection (LOD) and limit of quantification (LOQ) for the Real-Q kit were 453 and 750 IU/mL, respectively. The conversion factor from EBV genomic copies to IU was 0.62. The linear range of the assay was from 750 to 10⁶ IU/mL. Viral load values measured with the Real-Q assay were on average 0.54 log₁₀ copies/mL higher than those measured with the artus assay. The Real-Q assay offered good analytical performance for EBV DNA quantification in WB.

  8. The impact of targeting repetitive BamHI-W sequences on the sensitivity and precision of EBV DNA quantification.

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    Armen Sanosyan

    Full Text Available Viral load monitoring and early Epstein-Barr virus (EBV DNA detection are essential in routine laboratory testing, especially in preemptive management of Post-transplant Lymphoproliferative Disorder. Targeting the repetitive BamHI-W sequence was shown to increase the sensitivity of EBV DNA quantification, but the variability of BamHI-W reiterations was suggested to be a source of quantification bias. We aimed to assess the extent of variability associated with BamHI-W PCR and its impact on the sensitivity of EBV DNA quantification using the 1st WHO international standard, EBV strains and clinical samples.Repetitive BamHI-W- and LMP2 single- sequences were amplified by in-house qPCRs and BXLF-1 sequence by a commercial assay (EBV R-gene™, BioMerieux. Linearity and limits of detection of in-house methods were assessed. The impact of repeated versus single target sequences on EBV DNA quantification precision was tested on B95.8 and Raji cell lines, possessing 11 and 7 copies of the BamHI-W sequence, respectively, and on clinical samples.BamHI-W qPCR demonstrated a lower limit of detection compared to LMP2 qPCR (2.33 log10 versus 3.08 log10 IU/mL; P = 0.0002. BamHI-W qPCR underestimated the EBV DNA load on Raji strain which contained fewer BamHI-W copies than the WHO standard derived from the B95.8 EBV strain (mean bias: - 0.21 log10; 95% CI, -0.54 to 0.12. Comparison of BamHI-W qPCR versus LMP2 and BXLF-1 qPCR showed an acceptable variability between EBV DNA levels in clinical samples with the mean bias being within 0.5 log10 IU/mL EBV DNA, whereas a better quantitative concordance was observed between LMP2 and BXLF-1 assays.Targeting BamHI-W resulted to a higher sensitivity compared to LMP2 but the variable reiterations of BamHI-W segment are associated with higher quantification variability. BamHI-W can be considered for clinical and therapeutic monitoring to detect an early EBV DNA and a dynamic change in viral load.

  9. The impact of targeting repetitive BamHI-W sequences on the sensitivity and precision of EBV DNA quantification.

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    Sanosyan, Armen; Fayd'herbe de Maudave, Alexis; Bollore, Karine; Zimmermann, Valérie; Foulongne, Vincent; Van de Perre, Philippe; Tuaillon, Edouard

    2017-01-01

    Viral load monitoring and early Epstein-Barr virus (EBV) DNA detection are essential in routine laboratory testing, especially in preemptive management of Post-transplant Lymphoproliferative Disorder. Targeting the repetitive BamHI-W sequence was shown to increase the sensitivity of EBV DNA quantification, but the variability of BamHI-W reiterations was suggested to be a source of quantification bias. We aimed to assess the extent of variability associated with BamHI-W PCR and its impact on the sensitivity of EBV DNA quantification using the 1st WHO international standard, EBV strains and clinical samples. Repetitive BamHI-W- and LMP2 single- sequences were amplified by in-house qPCRs and BXLF-1 sequence by a commercial assay (EBV R-gene™, BioMerieux). Linearity and limits of detection of in-house methods were assessed. The impact of repeated versus single target sequences on EBV DNA quantification precision was tested on B95.8 and Raji cell lines, possessing 11 and 7 copies of the BamHI-W sequence, respectively, and on clinical samples. BamHI-W qPCR demonstrated a lower limit of detection compared to LMP2 qPCR (2.33 log10 versus 3.08 log10 IU/mL; P = 0.0002). BamHI-W qPCR underestimated the EBV DNA load on Raji strain which contained fewer BamHI-W copies than the WHO standard derived from the B95.8 EBV strain (mean bias: - 0.21 log10; 95% CI, -0.54 to 0.12). Comparison of BamHI-W qPCR versus LMP2 and BXLF-1 qPCR showed an acceptable variability between EBV DNA levels in clinical samples with the mean bias being within 0.5 log10 IU/mL EBV DNA, whereas a better quantitative concordance was observed between LMP2 and BXLF-1 assays. Targeting BamHI-W resulted to a higher sensitivity compared to LMP2 but the variable reiterations of BamHI-W segment are associated with higher quantification variability. BamHI-W can be considered for clinical and therapeutic monitoring to detect an early EBV DNA and a dynamic change in viral load.

  10. Analysis of DNA topology of EBV minichromosomes in HEK 293 cells.

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    Alicia Castán

    Full Text Available Simian Virus 40 (SV40 and Epstein-Barr Virus (EBV are frequently used as model systems to study DNA replication. Their genomes are both circular duplex DNAs organized in a single replicon where replication initiates at a precise site upon binding of a specific protein: the large tumor (T antigen for SV40 and the Epstein-Barr Nuclear Antigen 1 (EBNA-1 for EBV. Despite the abundant information available on the genetics and biochemistry of the replication process in these systems, little is known about the changes in DNA topology that take place as molecules are transfected into eukaryotic cells, assembled into chromatin and bind initiator proteins to start replication. Here we used high-resolution two-dimensional agarose gel electrophoresis to demonstrate that in Human Embryonic Kidney (HEK 293 cells, minichromosomes of almost the same mass carrying either the SV40 or the EBV replication origin showed similar topological features. The patterns were very similar regardless of the initiator proteins. We also showed that in a hybrid minichromosome, pEco3'Δ, that initiates replication from the SV40 origin, the presence of EBNA-1 and its putative binding to the EBV "family of repeats" induces no significant topological change. These observations challenge the idea that binding of EBNA-1 to oriP could induce negative supercoiling and favor a model suggesting that it binds to oriP in a two-step process where only the second step causes structural changes in a transient cell cycle specific manner.

  11. DNA Damage Signaling Is Induced in the Absence of Epstein-Barr Virus (EBV) Lytic DNA Replication and in Response to Expression of ZEBRA.

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    Wang'ondu, Ruth; Teal, Stuart; Park, Richard; Heston, Lee; Delecluse, Henri; Miller, George

    2015-01-01

    Epstein Barr virus (EBV), like other oncogenic viruses, modulates the activity of cellular DNA damage responses (DDR) during its life cycle. Our aim was to characterize the role of early lytic proteins and viral lytic DNA replication in activation of DNA damage signaling during the EBV lytic cycle. Our data challenge the prevalent hypothesis that activation of DDR pathways during the EBV lytic cycle occurs solely in response to large amounts of exogenous double stranded DNA products generated during lytic viral DNA replication. In immunofluorescence or immunoblot assays, DDR activation markers, specifically phosphorylated ATM (pATM), H2AX (γH2AX), or 53BP1 (p53BP1), were induced in the presence or absence of viral DNA amplification or replication compartments during the EBV lytic cycle. In assays with an ATM inhibitor and DNA damaging reagents in Burkitt lymphoma cell lines, γH2AX induction was necessary for optimal expression of early EBV genes, but not sufficient for lytic reactivation. Studies in lytically reactivated EBV-positive cells in which early EBV proteins, BGLF4, BGLF5, or BALF2, were not expressed showed that these proteins were not necessary for DDR activation during the EBV lytic cycle. Expression of ZEBRA, a viral protein that is necessary for EBV entry into the lytic phase, induced pATM foci and γH2AX independent of other EBV gene products. ZEBRA mutants deficient in DNA binding, Z(R183E) and Z(S186E), did not induce foci of pATM. ZEBRA co-localized with HP1β, a heterochromatin associated protein involved in DNA damage signaling. We propose a model of DDR activation during the EBV lytic cycle in which ZEBRA induces ATM kinase phosphorylation, in a DNA binding dependent manner, to modulate gene expression. ATM and H2AX phosphorylation induced prior to EBV replication may be critical for creating a microenvironment of viral and cellular gene expression that enables lytic cycle progression.

  12. Screening for PTLD in lung and heart-lung transplant recipients by measuring EBV DNA load in bronchoalveolar lavage fluid using real time PCR.

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    Michelson, Peter; Watkins, Bradley; Webber, Steven A; Wadowsky, Robert; Michaels, Marian G

    2008-06-01

    Pediatric L-HLTx recipients are at risk for developing PTLD with the lung being a primary site of disease. We hypothesized that BALF is a better sample than peripheral blood for measuring EBV DNA load in this high-risk population. Archived BALF specimens from pediatric L-HLTx recipients with and without PTLD were assayed for EBV DNA load using a quantitative real time TaqMan PCR assay. These values were compared with values determined in peripheral blood by a competitive PCR assay. Fifty-five BALF specimens from 16 L-HLTx patients were evaluated. Three patients with PTLD had mean BALF EBV DNA load values almost 50-fold higher than subjects without PTLD (4.6 x 10(5) copies/mL vs. 1.0 x 10(4) copies/mL). Patients who were EBV seronegative pretransplantation (i.e., high risk for PTLD) had elevated EBV DNA load values vs. patients who were EBV seropositive pretransplantation, regardless of the diagnosis of PTLD (mean values of 3.2 x 10(5) copies/mL vs. 1.1 x 10(4) copies/mL). Lastly, BALF analysis identified all subjects with PTLD, whereas peripheral blood analysis identified only one of these cases. Therefore, it can be concluded that monitoring EBV DNA load in BALF following L-HLTx facilitates detection of PTLD in high-risk patients and may be superior to peripheral blood assays.

  13. Quantificazione mediante PCR dell’EBV-DNA da biopsie cutanee di pazienti con linfomi cutanei primitivi (micosi fungoide e sindrome di Sèzary

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    Chiara Merlino

    2007-06-01

    Full Text Available Mycosis fungoides (MF, the most indolent form of CTCL, originates from a clonal expansion of epidermotropic helper/memory T cells. Sezary syndrome (SS is a rare primay epidermotropic cutaneous T-cell lymphoma in leukemic. The aetiopathogenesis of MF and SS remains obscure despite several investigations. Infectious, environmental and genetic factors have been implicated as potential aetiological agents. The studies investigating the role of EBV in CTCL present conflicting results. The different sensitivities of the technical methods used in the evaluation of the presence of viral DNA or virus-related antigens make comparison of the results difficult. The aim of this study was to retrospectively evaluate the EBV-DNA load in skin biopsies from MF and SS patients by a highly sensitive (1-10 EBV-DNA copies/reaction quantitative-competitive PCR (QC-PCR developed in our lab to better asses the relationship between EBV and CTCL. Skin biopsies were obtained from 21 MF and 10 SS patients; skin biopsies from a 8 patients with inflammatory skin disease were used as controls. EBV-DNA was detected in 70% of biopsies from SS patients vs. 0% of MF patients. No control patients resulted EBV-DNA positive, as expected. In addition, in SS patients, the survival from diagnosis is lesser in EBV-positive patients vs.EBV-negative patients even if not statistically significant.We are going to investigate the presence of EBV-DNA in peripheral blood of a larger number of patients and to evaluate the pattern of viral genes expression, to better assess the aetiopathogenetical role of EB virus in this kind of neoplasies.

  14. Induction of Epstein-Barr virus (EBV) lytic cycle in vitro causes lipid peroxidation, protein oxidation and DNA damage in lymphoblastoid B cell lines.

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    Gargouri, Bochra; Nasr, Rihab; Mseddi, Malek; Benmansour, Riadh; Lassoued, Saloua

    2011-07-01

    We investigated the oxidative modifications of lipids, proteins and DNA, potential molecular targets of oxidative stress, in two lymphoblastoid cell lines: B95-8 and Raji, after EBV lytic cycle induction. Conjugated dienes level was measured as biomarker of lipid peroxidation. Malondialdehyde adduct and protein carbonyl levels, as well as protein thiol levels were measured as biomarkers of protein oxidation. DNA fragmentation was evaluated as biomarker of DNA oxidation. After 48 h (peak of lytic cycle), a significant increase in conjugated dienes level was observed in B95-8 and Raji cell lines (p = 0.0001 and p = 0.019 respectively). Malondialdehyde adduct, protein carbonyl levels were increased in B95-8 and Raji cell lines after EBV lytic cycle induction as compared to controls (MDA-adduct: p = 0.008 and p = 0.006 respectively; Carbonyl: p = 0.003 and p = 0.0039 respectively). Proteins thiol levels were decreased by induction in B95-8 and Raji cell lines (p = 0.046; p = 0.002 respectively). DNA fragmentation was also detected in B95-8 and Raji cell lines after EBV lytic cycle induction as compared to controls. The results of this study demonstrate the presence of increased combined oxidative modifications in lipids, proteins in B95-8 and Raji cells lines after EBV lytic cycle induction. These results suggest that lipid peroxidation, protein oxidation and DNA fragmentation are generally induced during EBV lytic cycle induction and probably contribute to the cytopathic effect of EBV.

  15. Induction of epstein-barr virus (EBV lytic cycle in vitro causes lipid peroxidation, protein oxidation and DNA damage in lymphoblastoid B cell lines

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    benmansour Riadh

    2011-07-01

    Full Text Available Abstract Background We investigated the oxidative modifications of lipids, proteins and DNA, potential molecular targets of oxidative stress, in two lymphoblastoid cell lines: B95-8 and Raji, after EBV lytic cycle induction. Conjugated dienes level was measured as biomarker of lipid peroxidation. Malondialdehyde adduct and protein carbonyl levels, as well as protein thiol levels were measured as biomarkers of protein oxidation. DNA fragmentation was evaluated as biomarker of DNA oxidation. Results After 48 h (peak of lytic cycle, a significant increase in conjugated dienes level was observed in B95-8 and Raji cell lines (p = 0.0001 and p = 0.019 respectively. Malondialdehyde adduct, protein carbonyl levels were increased in B95-8 and Raji cell lines after EBV lytic cycle induction as compared to controls (MDA-adduct: p = 0.008 and p = 0.006 respectively; Carbonyl: p = 0.003 and p = 0.0039 respectively. Proteins thiol levels were decreased by induction in B95-8 and Raji cell lines (p = 0.046; p = 0.002 respectively. DNA fragmentation was also detected in B95-8 and Raji cell lines after EBV lytic cycle induction as compared to controls. Conclusion The results of this study demonstrate the presence of increased combined oxidative modifications in lipids, proteins in B95-8 and Raji cells lines after EBV lytic cycle induction. These results suggest that lipid peroxidation, protein oxidation and DNA fragmentation are generally induced during EBV lytic cycle induction and probably contribute to the cytopathic effect of EBV.

  16. Induction of epstein-barr virus (EBV) lytic cycle in vitro causes lipid peroxidation, protein oxidation and DNA damage in lymphoblastoid B cell lines

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    Gargouri, Bochra; Nasr, Rihab; Mseddi, Malek; benmansour, Riadh; Lassoued, saloua

    2011-01-01

    Abstract Background We investigated the oxidative modifications of lipids, proteins and DNA, potential molecular targets of oxidative stress, in two lymphoblastoid cell lines: B95-8 and Raji, after EBV lytic cycle induction. Conjugated dienes level was measured as biomarker of lipid peroxidation. Malondialdehyde adduct and protein carbonyl levels, as well as protein thiol levels were measured as biomarkers of protein oxidation. DNA fragmentation was evaluated as biomarker of DNA oxidation. Re...

  17. [DNA quantification of blood samples pre-treated with pyramidon].

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    Zhu, Chuan-Hong; Zheng, Dao-Li; Ni, Rao-Zhi; Wang, Hai-Sheng; Ning, Ping; Fang, Hui; Liu, Yan

    2014-06-01

    To study DNA quantification and STR typing of samples pre-treated with pyramidon. The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

  18. Prognostic value of serum Epstein-Barr virus antibodies in patients with nasopharyngeal carcinoma and undetectable pretreatment Epstein-Barr virus DNA.

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    Yao, Ji-Jin; Lin, Li; Jin, Ya-Nan; Wang, Si-Yang; Zhang, Wang-Jian; Zhang, Fan; Zhou, Guan-Qun; Cheng, Zhi-Bin; Qi, Zhen-Yu; Sun, Ying

    2017-08-01

    Epstein-Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC). Serum IgA antibodies against early antigen (EA-IgA) and viral capsid antigen (VCA-IgA) are the most commonly used to screen for NPC in endemic areas. However, the prognostic value of serum EA-IgA and VCA-IgA in patients with NPC is less clear. We hypothesize that serum EA-IgA and VCA-IgA levels have prognostic impact for survival outcomes in NPC patients with undetectable pretreatment EBV (pEBV) DNA. In this series, 334 patients with non-metastatic NPC and undetectable pEBV DNA were included. Serum EA-IgA and VCA-IgA were determined by ELISA. After analysis, serum EA-IgA and VCA-IgA loads correlated positively with T, N, and overall stage (all P 1:120 had significantly inferior 5-year progression-free survival (80.4% vs 89.6%, P = 0.025), distant metastasis-free survival (88.4% vs 94.8%, P = 0.050), and locoregional relapse-free survival (88.4% vs 95.6%, P = 0.023; log-rank test). Multivariable analyses revealed that N stage was the only independent prognostic factor (all P < 0.05), but the VCA-IgA became insignificant. Further analyses revealed that serum VCA-IgA was not an independent prognostic factor in early N (N0-1) or advanced N (N2-3) stage NPC. In summary, although both EA-IgA and VCA-IgA correlate strongly with TNM stage, our analyses do not suggest that these antibodies are prognostic biomarkers in patients with NPC and undetectable pEBV DNA. © 2017 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

  19. A simple procedure for the extraction of DNA from long-term formalin-preserved brain tissues for the detection of EBV by PCR.

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    Hassani, Asma; Khan, Gulfaraz

    2015-12-01

    Long-term formalin fixed brain tissues are potentially an important source of material for molecular studies. Ironically, very few protocols have been published describing DNA extraction from such material for use in PCR analysis. In our attempt to investigate the role of Epstein-Barr virus (EBV) in the pathogenesis of multiple sclerosis (MS), extracting PCR quality DNA from brain samples fixed in formalin for 2-22 years, proved to be very difficult and challenging. As expected, DNA extracted from these samples was not only of poor quality and quantity, but more importantly, it was frequently found to be non-amplifiable due to the presence of PCR inhibitors. Here, we describe a simple and reproducible procedure for extracting DNA using a modified proteinase K and phenol-chloroform methodology. Central to this protocol is the thorough pre-digestion washing of the tissues in PBS, extensive digestion with proteinase K in low SDS containing buffer, and using low NaCl concentration during DNA precipitation. The optimized protocol was used in extracting DNA from meninges of 26 MS and 6 non-MS cases. Although the quality of DNA from these samples was generally poor, small size amplicons (100-200 nucleotides) of the house-keeping gene, β-globin could be reliably amplified from all the cases. PCR for EBV revealed positivity in 35% (9/26) MS cases, but 0/6 non-MS cases. These findings indicate that the method described here is suitable for PCR detection of viral sequences in long-term formalin persevered brain tissues. Our findings also support a possible role for EBV in the pathogenesis of MS. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. The significance of Epstein Barr Virus (EBV & DNA Topoisomerase II alpha (DNA-Topo II alpha immunoreactivity in normal oral mucosa, Oral Epithelial Dysplasia (OED and Oral Squamous Cell Carcinoma (OSCC

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    Osman Mohamed M

    2008-11-01

    Full Text Available Abstract Background Head and neck cancer including oral cancer is considered to develop by accumulated genetic alterations and the major pathway is cancerization from lesions such as intraepithelial dysplasia in oral leukoplakia and erythroplakia. The relationship of proliferation markers with the grading of dysplasia is uncertain. The involvement of EBV in oral carcinogenesis is not fully understood. Aim The present study was designed to investigate the role of EBV and DNA Topoisomerase II∝ (DNA-Topo II∝ during oral carcinogenesis and to examine the prognostic significance of these protein expressions in OSCCs. Methods Using specific antibodies for EBV and DNA-Topo II∝, we examined protein expressions in archival lesion tissues from 16 patients with oral epithelial dysplasia, 22 oral squamous cell carcinoma and 20 normal oral mucosa by immunohistochemistry. Clinical information was obtained through the computerized retrospective database from the tumor registry. Results DNA-Topo II∝ was expressed in all examined specimens. Analysis of Variance ANOVA revealed highly significant difference (P 0.05 in inferior surface of tongue and in hard palatal tissues. Significant differences were observed between OEDs and NSE (P Conclusion EBV and DNA Topo II-αLI expression are possible indicators in oral carcinogenesis and may be valuable diagnostic and prognostic indices in oral carcinoma.

  1. The impact of EBV hybridization in clinical and biological medicine.

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    Morinet, F

    1989-01-01

    Hybridization technology allows the detection of EBV (Epstein-Barr Virus) DNA in lymphoproliferative diseases in immunosuppressed patients. Moreover, in HIV infected patients, LIP (Lymphocytic Interstitial Pneumonitis), hairy leucoplasia and lymphoma are associated with EBV by DNA hybridization. These procedures are also useful for understanding interactions between EBV and epithelial cells.

  2. Detection of EBV-DNA in serum samples of an immunosuppressed child during a three years follow-up: association of clinical and PCR data with active infection Detecção de EBV-DNA em amostras de soro de criança imunodeprimida durante três anos de seguimento: associação de dados clínicos e de PCR com a infecção ativa

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    Thelma Suely Okay

    2005-04-01

    Full Text Available Twenty-four whole blood and serum samples were drawn from an eight year-old heart transplant child during a 36 months follow-up. EBV serology was positive for VCA-IgM and IgG, and negative for EBNA-IgG at the age of five years old when the child presented with signs and symptoms suggestive of acute infectious mononucleosis. After 14 months, serological parameters were: positive VCA-IgG, EBNA-IgG and negative VCA-IgM. This serological pattern has been maintained since then even during episodes suggestive of EBV reactivation. PCR amplified a specific DNA fragment from the EBV gp220 (detection limit of 100 viral copies. All twenty-four whole blood samples yielded positive results by PCR, while 12 out of 24 serum samples were positive. We aimed at analyzing whether detection of EBV-DNA in serum samples by PCR was associated with overt disease as stated by the need of antiviral treatment and hospitalization. Statistical analysis showed agreement between the two parameters evidenced by the Kappa test (value 0.750; p Vinte e quatro amostras de sangue total e de soro foram colhidas durante seguimento por 36 meses de criança de oito anos de idade, imunodeprimida devido a transplante cardíaco. O paciente apresentou VCA-IgG e IgM positivos e EBNA-IgG negativo aos cinco anos de idade quando foi diagnosticada mononucleose infecciosa. Quatorze meses depois o VCA-IgG e o EBNA-IgG eram positivos e o VCA-IgM negativo. Este padrão sorológico persiste desde aquela época mesmo durante episódios sugestivos de reativação. As amostras de sangue total e de soro foram analisadas pela Reação em Cadeia da Polimerase (PCR que amplificou fragmento oriundo da gp220 do EBV (detecção de 100 cópias virais. Todas as 24 amostras de sangue total e 12 amostras de soro foram positivas por PCR. Com o objetivo de verificar se a detecção de DNA do EBV em soro estaria associada à reativação da doença, os resultados de PCR foram analisados em relação à necessidade

  3. EBV CHRONIC INFECTIONS

    Directory of Open Access Journals (Sweden)

    Eligio Pizzigallo

    2010-08-01

    Full Text Available The infection from Epstein-Barr virus (EBV or virus of infectious mononucleosis, together with other herpesviruses’ infections, represents a prototype of persistent viral infections characterized by the property of the latency. Although the reactivations of the latent infection are associated with the resumption of the viral replication and eventually with the “shedding”, it is still not clear if this virus can determine chronic infectious diseases, more or less evolutive. These diseases could include some pathological conditions actually defined as “idiopathic”and characterized by the “viral persistence” as the more credible pathogenetic factor. Among the so-called idiopathic syndromes, the “chronic fatigue syndrome” (CFS aroused a great interest around the eighties of the last century when, just for its relationship with EBV, it was called “chronic mononucleosis” or “chronic EBV infection”. Today CFS, as defined in 1994 by the CDC of Atlanta (USA, really represents a multifactorial syndrome characterized by a chronic course, where reactivation and remission phases alternate, and by a good prognosis. The etiopathogenetic role of EBV is demonstrated only in a well-examined subgroup of patients, while in most of the remaining cases this role should be played by other infectious agents - able to remain in a latent or persistent way in the host – or even by not infectious agents (toxic, neuroendocrine, methabolic, etc.. However, the pathogenetic substrate of the different etiologic forms seems to be the same, much probably represented by the oxidative damage due to the release of pro-inflammatory cytokines as a response to the triggering event (infectious or not infectious. Anyway, recently the scientists turned their’s attention to the genetic predisposition of the subjects affected by the syndrome, so that in the last years the genetic studies, together with those of molecular biology, received a great impulse

  4. On-Chip integration of sample pretreatment and Multiplex polymerase chain reaction (PCR) for DNA analysis

    DEFF Research Database (Denmark)

    Brivio, Monica; Snakenborg, Detlef; Søgaard, E.

    2008-01-01

    In this paper we present a modular lab-on-a-chip system for integrated sample pre-treatment (PT) by magnetophoresis and DNA amplification by polymerase chain reaction (PCR). It consists of a polymer-based microfluidic chip mounted on a custom-made thermocycler (Figure 1) and includes a simple...

  5. The Epstein-Barr virus (EBV)-encoded protein kinase, EBV-PK, but not the thymidine kinase (EBV-TK), is required for ganciclovir and acyclovir inhibition of lytic viral production.

    Science.gov (United States)

    Meng, Qiao; Hagemeier, Stacy R; Fingeroth, Joyce D; Gershburg, Edward; Pagano, Joseph S; Kenney, Shannon C

    2010-05-01

    Ganciclovir (GCV) and acyclovir (ACV) are guanine nucleoside analogues that inhibit lytic herpesvirus replication. GCV and ACV must be monophosphorylated by virally encoded enzymes to be converted into nucleotides and incorporated into viral DNA. However, whether GCV and/or ACV phosphorylation in Epstein-Barr virus (EBV)-infected cells is mediated primarily by the EBV-encoded protein kinase (EBV-PK), the EBV-encoded thymidine kinase (EBV-TK), or both is controversial. To examine this question, we constructed EBV mutants containing stop codons in either the EBV-PK or EBV-TK open reading frame and selected for stable 293T clones latently infected with wild-type EBV or each of the mutant viruses. Cells were induced to the lytic form of viral replication with a BZLF1 expression vector in the presence and absence of various doses of GCV and ACV, and infectious viral titers were determined by a green Raji cell assay. As expected, virus production in wild-type EBV-infected 293T cells was inhibited by both GCV (50% inhibitory concentration [IC(50)] = 1.5 microM) and ACV (IC(50) = 4.1 microM). However, the EBV-PK mutant (which replicates as well as the wild-type (WT) virus in 293T cells) was resistant to both GCV (IC(50) = 19.6 microM) and ACV (IC(50) = 36.4 microM). Expression of the EBV-PK protein in trans restored GCV and ACV sensitivity in cells infected with the PK mutant virus. In contrast, in 293T cells infected with the TK mutant virus, viral replication remained sensitive to both GCV (IC(50) = 1.2 microM) and ACV (IC(50) = 2.8 microM), although susceptibility to the thymine nucleoside analogue, bromodeoxyuridine, was reduced. Thus, EBV-PK but not EBV-TK mediates ACV and GCV susceptibilities.

  6. EBV-positive mucocutaneous ulcer in organ transplant recipients: a localized indolent posttransplant lymphoproliferative disorder.

    Science.gov (United States)

    Hart, Melissa; Thakral, Beenu; Yohe, Sophia; Balfour, Henry H; Singh, Charanjeet; Spears, Michael; McKenna, Robert W

    2014-11-01

    Epstein-Barr virus (EBV)-positive mucocutaneous ulcer (EBV MCU) is a B-cell lymphoproliferative disorder occurring in elderly or iatrogenic immunocompromised patients. It has not been reported in solid organ transplant recipients. We observed 7 patients with EBV MCU in a cohort of 70 transplant recipients with EBV posttransplant lymphoproliferative disorder (PTLD). Transplants included: 5 renal, 1 heart, and 1 lung. Median patient age was 61; 5 were male. EBV MCU was observed in oral mucosa in 4 and gastrointestinal tract in 3. Duration of immunosuppressive therapy before EBV MCU was 0.6 to 13 years. Ulcers were undermined by inflammatory cells and polymorphic or monomorphic large cell lymphoproliferation. Reed-Sternberg-like cells were present in 5/7. Large B cells were CD20, CD30, and EBV-encoded RNA positive in all cases. Diagnosis in 3 recent patients was EBV MCU; 4 patients diagnosed before familiarity with EBV MCU were classified as monomorphic large cell (n=3) and polymorphic (n=1) PTLD. None of the patients had EBV DNA in their blood (<1000 copies/mL) at diagnosis or follow-up versus 35/44 transplant patients with systemic PTLD (P<0.001). All lesions resolved with reduced immunosuppression (7/7), change in immunosuppression (2/7), and rituximab (3/7). Five patients are living: 4 healthy, 1 awaiting second renal transplant. Two patients died 3 and 5 years after resolution of EBV MCU. No patient recurred with EBV MCU or other PTLDs. EBV MCU mimics more aggressive categories of PTLD but lacks EBV DNA in blood, which may be a useful distinguishing feature. Lesions are likely to resolve with conservative management. Awareness of EBV MCU in the posttransplant setting is necessary for appropriate diagnosis and treatment.

  7. Impact of Plasma Epstein-Barr Virus-DNA and Tumor Volume on Prognosis of Locally Advanced Nasopharyngeal Carcinoma

    Directory of Open Access Journals (Sweden)

    Meng Chen

    2015-01-01

    Full Text Available This retrospective study aims to examine the association of plasma Epstein-Barr virus- (EBV- DNA levels with the tumor volume and prognosis in patients with locally advanced nasopharyngeal carcinoma (NPC. A total of 165 patients with newly diagnosed locally advanced NPC were identified from September 2011 to July 2012. EBV-DNA was detected using fluorescence quantitative polymerase chain reaction (PCR amplification. The tumor volume was calculated by the systematic summation method of computer software. The median copy number of plasma EBV-DNA before treatment was 3790 copies/mL. The median gross tumor volume of the primary nasopharyngeal tumor (GTVnx, the lymph node lesions (GTVnd, and the total GTV before treatment were 72.46, 23.26, and 106.25 cm3, respectively; the EBV-DNA levels were significantly correlated with the GTVnd and the total GTV (P<0.01. The 2-year overall survival (OS rates in patients with positive and negative pretreatment plasma EBV-DNA were 100% and 98.4% (P=1.000, and the disease-free survival (DFS rates were 94.4% and 80.8% (P=0.044, respectively. These results indicate that high pretreatment plasma EBV-DNA levels in patients with locally advanced NPC are associated with the degree of lymph node metastasis, tumor burden, and poor prognosis.

  8. Long-Term Prognostic Effects of Plasma Epstein-Barr Virus DNA by Minor Groove Binder-Probe Real-Time Quantitative PCR on Nasopharyngeal Carcinoma Patients Receiving Concurrent Chemoradiotherapy

    International Nuclear Information System (INIS)

    Lin, J.-C.; Wang, W.-Y.; Liang, W.-M.; Chou, H.-Y.; Jan, J.-S.; Jiang, R.-S.; Wang, J.-Y.; Twu, C.-W.; Liang, K.-L.; Chao, Jeffrey; Shen, W.-C.

    2007-01-01

    Purpose: To evaluate the long-term prognostic impact of plasma Epstein-Barr virus (EBV) DNA concentration measured by real-time quantitative polymerase chain reaction (RTQ-PCR) in nasopharyngeal carcinoma (NPC) patients receiving concurrent chemoradiotherapy (CCRT). Methods and Materials: Epstein-Barr virus DNA was retrospectively measured from stock plasma of 152 biopsy-proven NPC patients with Stage II-IV (M0) disease with a RTQ-PCR using the minor groove binder-probe. All patients received CCRT with a median follow-up of 78 months. We divided patients into three subgroups: (1) low pretreatment EBV DNA (<1,500 copies/mL) and undetectable posttreatment EBV DNA (pre-L/post-U) (2) high pretreatment EBV DNA (≥1,500 copies/mL) and undetectable posttreatment EBV DNA (pre-H/post-U), and (3) low or high pretreatment EBV DNA and detectable posttreatment EBV DNA (pre-L or H/post-D) for prognostic analyses. Results: Epstein-Barr virus DNA (median concentration, 573 copies/mL; interquartile range, 197-3,074) was detected in the pretreatment plasma of 94.1% (143/152) of patients. After treatment, plasma EBV DNA decreased or remained 0 for all patients and was detectable in 31 patients (20.4%) with a median concentration 0 copy/mL (interquartile range, 0-0). The 5-year overall survival rates of the pre-L/post-U, pre-H/post-U, and pre-L or H/post-D subgroups were 87.2%, 71.0%, and 38.7%, respectively (p < 0.0001). The relapse-free survival showed similar results with corresponding rates of 85.6%, 75.9%, and 26.9%, respectively (p < 0.0001). Multivariate Cox analysis confirmed the superior effects of plasma EBV DNA compared to other clinical parameters in prognosis prediction. Conclusion: Plasma EBV DNA is the most valuable prognostic factor for NPC. More chemotherapy should be considered for patients with persistently detectable EBV DNA after CCRT

  9. EBV CHRONIC INFECTIONS

    Directory of Open Access Journals (Sweden)

    Delia Racciatti

    2010-02-01

    Full Text Available

    The infection from Epstein-Barr virus (EBV or virus of infectious mononucleosis, together with other herpesviruses’ infections, represents a prototype of persistent viral infections characterized by the property of the latency. Although the reactivations of the latent infection are associated with the resumption of the viral replication and eventually with the “shedding”, it is still not clear if this virus can determine chronic infectious diseases, more or less evolutive. These diseases could include some pathological conditions actually defined as “idiopathic”and characterized by the “viral persistence” as the more credible pathogenetic factor. Among the so-called idiopathic syndromes, the “chronic fatigue syndrome” (CFS aroused a great interest around the eighties of the last century when, just for its relationship with EBV, it was called “chronic mononucleosis” or “chronic EBV infection”.

    Today CFS, as defined in 1994 by the CDC of Atlanta (USA, really represents a multifactorial syndrome characterized by a chronic course, where reactivation and remission phases alternate, and by a good prognosis

  10. On-Chip integration of sample pretreatment and Multiplex polymerase chain reaction (PCR) for DNA analysis

    DEFF Research Database (Denmark)

    Brivio, Monica; Snakenborg, Detlef; Søgaard, E.

    2008-01-01

    In this paper we present a modular lab-on-a-chip system for integrated sample pre-treatment (PT) by magnetophoresis and DNA amplification by polymerase chain reaction (PCR). It consists of a polymer-based microfluidic chip mounted on a custom-made thermocycler (Figure 1) and includes a simple...... and efficient method for switching the liquid flow between the PT and PCR chamber. Purification of human genomic DNA from EDTA-treated blood and multiplex PCR were successfully carried out on-chip using the developed lab-on-a-chip system....

  11. Prognostic value of pre-treatment human papilloma virus DNA status in cervical cancer.

    Science.gov (United States)

    Chong, Gun Oh; Lee, Yoon Hee; Han, Hyung Soo; Lee, Hyun Jung; Park, Ji Young; Hong, Dae Gy; Lee, Yoon Soon; Cho, Young Lae

    2018-01-01

    Although the relationship between human papilloma virus (HPV) and cervical cancer is well established, the prognostic value of HPV status has not been determined, largely because previous studies have yielded conflicting results. This study aimed to investigate the prognostic value of pre-treatment HPV DNA for predicting tumor recurrence in cervical cancer. The study included 248 eligible patients who provided cervical cell specimens for HPV genotyping before surgery or concurrent chemoradiotherapy (CCRT). Of these 248 patients, 108 were treated with radical hysterectomy for International Federation of Gynecology and Obstetrics (FIGO) stage IB1-IIA cervical cancer, and 140 were treated with CCRT for FIGO stage IB2-IV cervical cancer. HPV 16 and 18 were the two most common HPV types detected, with prevalence rates of 52.4% and 12.5%, respectively. The pre-treatment HPV DNA test showed that 18.5% of cervical cancers were HPV negative. Multivariate analysis showed that HPV negativity was associated with poorer disease-free survival (DFS) than HPV-positive status (hazard ratio [HR], 3.97; 95% confidence interval [CI], 1.84-8.58; p=0.0005), and patients with HPV 16-positive cancers had better DFS (HR, 0.41; 95% CI, 0.23-0.72; p=0.0019). In the surgery group, only HPV 16 positivity was significantly correlated with DFS (HR, 0.34; 95% CI, 0.12-0.96; p=0.0416). In the CCRT group, only HPV negativity was significantly correlated with DFS (HR, 3.75; 95% CI, 1.78-7.90; p=0.0005). Pre-treatment HPV DNA status may be a useful prognostic biomarker in cervical cancer. The presence of HPV 16 DNA was associated with better DFS, and HPV negativity was associated with worse DFS. However, larger sample sizes and more comprehensive studies are required to verify our findings. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Increased levels of unscheduled DNA synthesis in UV-irradiated human fibroblasts pretreated with sodium butyrate

    International Nuclear Information System (INIS)

    Williams, J.I.; Friedberg, E.C.

    1982-01-01

    Pretreatment of growing normal and xeroderma pigmentosum (XP) human fibroblasts with sodium butyrate at concentrations of 5-20 mM results in increased levels of DNA repair synthesis measured by autoradiography after exposure of the cells to 254 nm UV radiation in the fluence range 0-25 J/m 2 . The phenomenon manifests as an increased extent and an increased initial rate of unscheduled DNA synthesis (UDS). This experimental result is not due to an artifact of autoradiography related to cell size. Xeroderma pigmentosum cells from complementation groups A, C, D and E and XP variant cells all exhibit increases in the levels of UV-induced UDS in response to sodium butyrate proportional to those observed with normal cells. These UDS increases associated with butyrate pretreatment correlate with demonstrable changes in intracellular thymidine pool size and suggest that sodium butyrate enhances uptake of exogenous radiolabeled thymidine during UV-induced repair synthesis by reducing endogenous levels of thymidine. (author)

  13. Unexpected instability of family of repeats (FR, the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems.

    Directory of Open Access Journals (Sweden)

    Teru Kanda

    Full Text Available A group of repetitive sequences, known as the Family of Repeats (FR, is a critical cis-acting sequence required for EBV latent infection. The FR sequences are heterogeneous among EBV strains, and they are sometimes subject to partial deletion when subcloned in E. coli-based cloning vectors. However, the FR stability in EBV-BAC (bacterial artificial chromosome system has never been investigated. We found that the full length FR of the Akata strain EBV was not stably maintained in a BAC vector. By contrast, newly obtained BAC clones of the B95-8 strain of EBV stably maintained the full length FR during recombinant virus production and B-cell transformation. Investigation of primary DNA sequences of Akata-derived EBV-BAC clones indicates that the FR instability is most likely due to a putative secondary structure of the FR region. We conclude that the FR instability in EBV-BAC clones can be a pitfall in E. coli-mediated EBV genetics.

  14. Leflunomide/teriflunomide inhibit Epstein-Barr virus (EBV)- induced lymphoproliferative disease and lytic viral replication.

    Science.gov (United States)

    Bilger, Andrea; Plowshay, Julie; Ma, Shidong; Nawandar, Dhananjay; Barlow, Elizabeth A; Romero-Masters, James C; Bristol, Jillian A; Li, Zhe; Tsai, Ming-Han; Delecluse, Henri-Jacques; Kenney, Shannon C

    2017-07-04

    EBV infection causes mononucleosis and is associated with specific subsets of B cell lymphomas. Immunosuppressed patients such as organ transplant recipients are particularly susceptible to EBV-induced lymphoproliferative disease (LPD), which can be fatal. Leflunomide (a drug used to treat rheumatoid arthritis) and its active metabolite teriflunomide (used to treat multiple sclerosis) inhibit de novo pyrimidine synthesis by targeting the cellular dihydroorotate dehydrogenase, thereby decreasing T cell proliferation. Leflunomide also inhibits the replication of cytomegalovirus and BK virus via both "on target" and "off target" mechanisms and is increasingly used to treat these viruses in organ transplant recipients. However, whether leflunomide/teriflunomide block EBV replication or inhibit EBV-mediated B cell transformation is currently unknown. We show that teriflunomide inhibits cellular proliferation, and promotes apoptosis, in EBV-transformed B cells in vitro at a clinically relevant dose. In addition, teriflunomide prevents the development of EBV-induced lymphomas in both a humanized mouse model and a xenograft model. Furthermore, teriflunomide inhibits lytic EBV infection in vitro both by preventing the initial steps of lytic viral reactivation, and by blocking lytic viral DNA replication. Leflunomide/teriflunomide might therefore be clinically useful for preventing EBV-induced LPD in patients who have high EBV loads yet require continued immunosuppression.

  15. EBV and vitamin D status in relapsing-remitting multiple sclerosis patients with a unique cytokine signature.

    Science.gov (United States)

    Nejati, Ahmad; Shoja, Zabihollah; Shahmahmoodi, Shohreh; Tafakhori, Abbas; Mollaei-Kandelous, Yaghoub; Rezaei, Farhad; Hamid, Kabir Magaji; Mirshafiey, Abbas; Doosti, Rozita; Sahraian, Mohammad Ali; Mahmoudi, Mahmood; Shokri, Fazel; Emery, Vince; Marashi, Sayed Mahdi

    2016-04-01

    Multiple sclerosis, a debilitating autoimmune and inflammatory disease of the central nervous system, is associated with both infectious and non-infectious factors. We investigated the role of EBV infection, vitamin D level, and cytokine signature in MS patients. Molecular and serological assays were used to investigate immune biomarkers, vitamin D level, and EBV status in 83 patients with relapsing-remitting multiple sclerosis and 62 healthy controls. In total, 98.8 % of MS patients showed a history of EBV exposure compared to 88.6 % in the healthy group (p = 0.005). EBV DNA load was significantly higher in MS patients than healthy subjects (p vitamin D concentration and EBV load, but not EBNA-1 IgG antibody levels. Our data highlight biomarker correlates in MS patients together with a complex interplay between EBV replication and vitamin D levels.

  16. Prevention of EBV lymphoma development by oncolytic myxoma virus in a murine xenograft model of post-transplant lymphoproliferative disease

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Manbok, E-mail: manbok66@dankook.ac.kr [Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610 (United States); Rahman, Masmudur M. [Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610 (United States); Cogle, Christopher R. [Department of Hematology/Oncology, University of Florida, Gainesville, FL 32610 (United States); McFadden, Grant [Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610 (United States)

    2015-07-10

    Epstein–Barr virus (EBV) has been associated with a variety of epithelial and hematologic malignancies, including B-, T- and NK cell-lymphomas, Hodgkin's disease (HD), post-transplant lymphoproliferative diseases (LPDs), nasopharyngeal and gastric carcinomas, smooth muscle tumors, and HIV-associated lymphomas. Currently, treatment options for EBV-associated malignancies are limited. We have previously shown that myxoma virus specifically targets various human solid tumors and leukemia cells in a variety of animal models, while sparing normal human or murine tissues. Since transplant recipients of bone marrow or solid organs often develop EBV-associated post-transplant LPDs and lymphoma, myxoma virus may be of utility to prevent EBV-associated malignancies in immunocompromised transplant patients where treatment options are frequently limited. In this report, we demonstrate the safety and efficacy of myxoma virus purging as a prophylactic strategy for preventing post-transplant EBV-transformed human lymphomas, using a highly immunosuppressed mouse xenotransplantation model. This provides support for developing myxoma virus as a potential oncolytic therapy for preventing EBV-associated LPDs following transplantation of bone marrow or solid organ allografts. - Highlights: • Myxoma virus effectively infects and purges EBV lymphoma cells in vivo. • Oncolytic myxoma virus effectively eradicates oncogenic EBV tumorigenesis. • Ex vivo pre-treatment of myxoma virus can be effective as a preventive treatment modality for post-transplant lymphoproliferative diseases.

  17. CTCF prevents the epigenetic drift of EBV latency promoter Qp.

    Directory of Open Access Journals (Sweden)

    Italo Tempera

    2010-08-01

    Full Text Available The establishment and maintenance of Epstein-Barr Virus (EBV latent infection requires distinct viral gene expression programs. These gene expression programs, termed latency types, are determined largely by promoter selection, and controlled through the interplay between cell-type specific transcription factors, chromatin structure, and epigenetic modifications. We used a genome-wide chromatin-immunoprecipitation (ChIP assay to identify epigenetic modifications that correlate with different latency types. We found that the chromatin insulator protein CTCF binds at several key regulatory nodes in the EBV genome and may compartmentalize epigenetic modifications across the viral genome. Highly enriched CTCF binding sites were identified at the promoter regions upstream of Cp, Wp, EBERs, and Qp. Since Qp is essential for long-term maintenance of viral genomes in type I latency and epithelial cell infections, we focused on the role of CTCF in regulating Qp. Purified CTCF bound approximately 40 bp upstream of the EBNA1 binding sites located at +10 bp relative to the transcriptional initiation site at Qp. Mutagenesis of the CTCF binding site in EBV bacmids resulted in a decrease in the recovery of stable hygromycin-resistant episomes in 293 cells. EBV lacking the Qp CTCF site showed a decrease in Qp transcription initiation and a corresponding increase in Cp and Fp promoter utilization at 8 weeks post-transfection. However, by 16 weeks post-transfection, bacmids lacking CTCF sites had no detectable Qp transcription and showed high levels of histone H3 K9 methylation and CpG DNA methylation at the Qp initiation site. These findings provide direct genetic evidence that CTCF functions as a chromatin insulator that prevents the promiscuous transcription of surrounding genes and blocks the epigenetic silencing of an essential promoter, Qp, during EBV latent infection.

  18. CTCF Prevents the Epigenetic Drift of EBV Latency Promoter Qp

    Science.gov (United States)

    Tempera, Italo; Wiedmer, Andreas; Dheekollu, Jayaraju; Lieberman, Paul M.

    2010-01-01

    The establishment and maintenance of Epstein-Barr Virus (EBV) latent infection requires distinct viral gene expression programs. These gene expression programs, termed latency types, are determined largely by promoter selection, and controlled through the interplay between cell-type specific transcription factors, chromatin structure, and epigenetic modifications. We used a genome-wide chromatin-immunoprecipitation (ChIP) assay to identify epigenetic modifications that correlate with different latency types. We found that the chromatin insulator protein CTCF binds at several key regulatory nodes in the EBV genome and may compartmentalize epigenetic modifications across the viral genome. Highly enriched CTCF binding sites were identified at the promoter regions upstream of Cp, Wp, EBERs, and Qp. Since Qp is essential for long-term maintenance of viral genomes in type I latency and epithelial cell infections, we focused on the role of CTCF in regulating Qp. Purified CTCF bound ∼40 bp upstream of the EBNA1 binding sites located at +10 bp relative to the transcriptional initiation site at Qp. Mutagenesis of the CTCF binding site in EBV bacmids resulted in a decrease in the recovery of stable hygromycin-resistant episomes in 293 cells. EBV lacking the Qp CTCF site showed a decrease in Qp transcription initiation and a corresponding increase in Cp and Fp promoter utilization at 8 weeks post-transfection. However, by 16 weeks post-transfection, bacmids lacking CTCF sites had no detectable Qp transcription and showed high levels of histone H3 K9 methylation and CpG DNA methylation at the Qp initiation site. These findings provide direct genetic evidence that CTCF functions as a chromatin insulator that prevents the promiscuous transcription of surrounding genes and blocks the epigenetic silencing of an essential promoter, Qp, during EBV latent infection. PMID:20730088

  19. Biopsy-proven case of Epstein-Barr virus (EBV)-associated vasculitis of the central nervous system.

    Science.gov (United States)

    Kano, Kohei; Katayama, Takayuki; Takeguchi, Shiori; Asanome, Asuka; Takahashi, Kae; Saito, Tsukasa; Sawada, Jun; Saito, Masato; Anei, Ryogo; Kamada, Kyousuke; Miyokawa, Naoyuki; Nishihara, Hiroshi; Hasebe, Naoyuki

    2017-06-01

    A 75-year-old woman was admitted to our hospital with rapidly deteriorating consciousness disturbance. She had a 7-year history of rheumatoid arthritis (RA), which had been treated with methotrexate (MTX) and prednisolone. Brain T2-weighted MRI showed diffuse high-intensity lesions in the cerebral subcortical and deep white matter, bilateral basal ganglia and thalamus. A cerebrospinal fluid examination revealed elevated protein levels and positive Epstein-Barr virus (EBV) DNA. Human immunodeficiency virus was negative. Brain biopsy showed perivascular lymphocytic infiltration in the parenchyma and meninx with EBV-encoded small RNA (EBER). Since this case did not fulfill the criteria for chronic active EBV infection (CAEBV), she was diagnosed with Epstein-Barr virus (EBV)-associated vasculitis of the central nervous system. High-dose methylprednisolone, acyclovir, ganciclovir and foscarnet were not effective. Although EBV is a causative agent of infectious mononucleosis (IM), lymphomas and nasopharyngeal carcinomas, vasculitic pathology of the central nervous system with EBV reactivation in the elderly is rare. Immunosuppressive drugs such as steroids and MTX are widely used to treat autoimmune disorders, but may exacerbate the reactivation of EBV. This is the first case of biopsy-proven EBV-positive/HIV-negative vasculitis during the treatment of RA with MTX and steroids. This case indicates that EBV-associated vasculitis needs to be considered as a differential diagnosis of CNS vasculitis. © 2016 Japanese Society of Neuropathology.

  20. About Epstein-Barr Virus (EBV)

    Science.gov (United States)

    ... For Healthcare Providers Laboratory Testing References & Resources About Epstein-Barr Virus (EBV) Language: English (US) Español (Spanish) Recommend on ... On This Page Symptoms Transmission Diagnosis Prevention & Treatment Epstein-Barr virus (EBV), also known as human herpesvirus 4, is ...

  1. Plasma Epstein-Barr viral DNA complements TNM classification of nasopharyngeal carcinoma in the era of intensity-modulated radiotherapy.

    Science.gov (United States)

    Zhang, Lu; Tang, Lin-Quan; Chen, Qiu-Yan; Liu, Huai; Guo, Shan-Shan; Liu, Li-Ting; Guo, Ling; Mo, Hao-Yuan; Zhao, Chong; Guo, Xiang; Cao, Ka-Jia; Qian, Chao-Nan; Zeng, Mu-Sheng; Shao, Jian-Yong; Sun, Ying; Ma, Jun; Hong, Ming-Huang; Mai, Hai-Qiang

    2016-02-02

    The objective of this study is to verify the prognostic value of pretreatment plasma Epstein-Barr viral deoxyribonucleic acid (pEBV DNA) levels in nasopharyngeal carcinoma (NPC) patients to complement TNM classification based on the application of the intensity-modulated radiotherapy (IMRT) technique. In total, 1467 patients staged at I-IVa-b (M0) and treated with IMRT were retrospectively analyzed at our cancer center from January 2007 to December 2010. Patient survival among different stages and EBV DNA levels were compared. Outcome analyses of different stages and EBV DNA levels revealed that patients in stages II-III with low EBV DNA levels had similar survival as that of patients in stages IVa-b with low EBV DNA (5-yr overall survival (OS), 94.7% vs. 92.9% (P = 0.141), progression failure-free survival (PFS), 87.2% vs. 89.0% (P = 0.685), distant metastasis failure-free survival (DMFS), 93.5% vs. 92.4% (P = 0.394) and locoregional failure-free survival (LRFS), 93.8% vs. 96.3% (P = 0.523)). Conversely, patients in stages II-III with high EBV DNA had better survival than patients in stages IVa-b with high EBV DNA (5-yr OS, 82.7% vs. 71.7% (P = 0.001), PFS, 70.7% vs. 66.2% (P = 0.047), DMFS, 79.6% vs. 74.8% (P = 0.066) and LRFS, 89.3% vs. 87.6% (P = 0.425)) but poorer survival than patients in stages IVa-b with low EBV DNA (5-yr OS, 82.7% vs. 92.9% (P = 0.025), PFS, 70.7% vs. 89.0, (P < 0.001), DMFS, 79.6% vs. 92.4%, (P = 0.001), LRFS, 89.3% vs. 96.3%, (P = 0.022)). pEBV DNA is a strong prognostic factor for patients with NPC when complemented with TNM staging in the era of IMRT application.

  2. Heterogeneity of the Epstein-Barr Virus (EBV) Major Internal Repeat Reveals Evolutionary Mechanisms of EBV and a Functional Defect in the Prototype EBV Strain B95-8.

    Science.gov (United States)

    Ba Abdullah, Mohammed M; Palermo, Richard D; Palser, Anne L; Grayson, Nicholas E; Kellam, Paul; Correia, Samantha; Szymula, Agnieszka; White, Robert E

    2017-12-01

    Epstein-Barr virus (EBV) is a ubiquitous pathogen of humans that can cause several types of lymphoma and carcinoma. Like other herpesviruses, EBV has diversified through both coevolution with its host and genetic exchange between virus strains. Sequence analysis of the EBV genome is unusually challenging because of the large number and lengths of repeat regions within the virus. Here we describe the sequence assembly and analysis of the large internal repeat 1 of EBV (IR1; also known as the BamW repeats) for more than 70 strains. The diversity of the latency protein EBV nuclear antigen leader protein (EBNA-LP) resides predominantly within the exons downstream of IR1. The integrity of the putative BWRF1 open reading frame (ORF) is retained in over 80% of strains, and deletions truncating IR1 always spare BWRF1. Conserved regions include the IR1 latency promoter (Wp) and one zone upstream of and two within BWRF1. IR1 is heterogeneous in 70% of strains, and this heterogeneity arises from sequence exchange between strains as well as from spontaneous mutation, with interstrain recombination being more common in tumor-derived viruses. This genetic exchange often incorporates regions of Epstein-Barr virus (EBV) infects the majority of the world population but causes illness in only a small minority of people. Nevertheless, over 1% of cancers worldwide are attributable to EBV. Recent sequencing projects investigating virus diversity to see if different strains have different disease impacts have excluded regions of repeating sequence, as they are more technically challenging. Here we analyze the sequence of the largest repeat in EBV (IR1). We first characterized the variations in protein sequences encoded across IR1. In studying variations within the repeat of each strain, we identified a mutation in the main laboratory strain of EBV that impairs virus function, and we suggest that tumor-associated viruses may be more likely to contain DNA mixed from two strains. The

  3. The Effect of Antiretroviral Combination Treatment on Epstein-Barr Virus (EBV Genome Load in HIV-Infected Patients

    Directory of Open Access Journals (Sweden)

    Anna M. C. Friis

    2010-03-01

    Full Text Available We evaluated the effect of combination anti-retroviral treatment (cART on the host control of EBV infection in moderately immunosuppressed HIV-1 patients. Twenty HIV-1 infected individuals were followed for five years with repeated measurements of EBV DNA load in peripheral blood lymphocytes in relation to HIV-RNA titers and CD4+ cell counts. Individuals with optimal response, i.e. durable non-detectable HIV-RNA, showed a decline of EBV load to the level of healthy controls. Individuals with non-optimal HIV-1 control did not restore their EBV control. Long-lasting suppression of HIV-replication after early initiation of cART is a prerequisite for re-establishing the immune control of EBV.

  4. Hypermethylation of the tumor suppressor gene PRDM1/Blimp-1 supports a pathogenetic role in EBV-positive Burkitt lymphoma

    International Nuclear Information System (INIS)

    Zhang, T; Ma, J; Nie, K; Yan, J; Liu, Y; Bacchi, C E; Queiroga, E M; Gualco, G; Sample, J T; Orazi, A; Knowles, D M; Tam, W

    2014-01-01

    PRDM1/Blimp-1 is a tumor suppressor gene in the activated B-cell subtype of diffuse large B-cell lymphomas. Its inactivation contributes to pathogenesis in this setting by impairing terminal B-cell differentiation induced by constitutive nuclear factor-κB activation. The role of PRDM1 in Burkitt lymphoma (BL) lymphomagenesis is not known. Here we identified hypermethylation of the promoter region and exon 1 of PRDM1 in all six Epstein–Barr virus (EBV)-positive BL cell lines and 12 of 23 (52%) primary EBV-positive BL or BL-related cases examined, but in none of the EBV-negative BL cell lines or primary tumors that we assessed, implying a tumor suppressor role for PRDM1 specifically in EBV-associated BL. A direct induction of PRDM1 hypermethylation by EBV is unlikely, as PRDM1 hypermethylation was not observed in EBV-immortalized B lymphoblastoid cell lines. Treatment of EBV-positive BL cells with 5′ azacytidine resulted in PRDM1 induction associated with PRDM1 demethylation, consistent with transcriptional silencing of PRDM1 as a result of DNA methylation. Overexpression of PRDM1 in EBV-positive BL cell lines resulted in cell cycle arrest. Our results expand the spectrum of lymphoid malignancies in which PRDM1 may have a tumor suppressor role and identify an epigenetic event that likely contributes to the pathogenesis of BL

  5. Hypermethylation of the tumor suppressor gene PRDM1/Blimp-1 supports a pathogenetic role in EBV-positive Burkitt lymphoma.

    Science.gov (United States)

    Zhang, T; Ma, J; Nie, K; Yan, J; Liu, Y; Bacchi, C E; Queiroga, E M; Gualco, G; Sample, J T; Orazi, A; Knowles, D M; Tam, W

    2014-11-07

    PRDM1/Blimp-1 is a tumor suppressor gene in the activated B-cell subtype of diffuse large B-cell lymphomas. Its inactivation contributes to pathogenesis in this setting by impairing terminal B-cell differentiation induced by constitutive nuclear factor-κB activation. The role of PRDM1 in Burkitt lymphoma (BL) lymphomagenesis is not known. Here we identified hypermethylation of the promoter region and exon 1 of PRDM1 in all six Epstein-Barr virus (EBV)-positive BL cell lines and 12 of 23 (52%) primary EBV-positive BL or BL-related cases examined, but in none of the EBV-negative BL cell lines or primary tumors that we assessed, implying a tumor suppressor role for PRDM1 specifically in EBV-associated BL. A direct induction of PRDM1 hypermethylation by EBV is unlikely, as PRDM1 hypermethylation was not observed in EBV-immortalized B lymphoblastoid cell lines. Treatment of EBV-positive BL cells with 5' azacytidine resulted in PRDM1 induction associated with PRDM1 demethylation, consistent with transcriptional silencing of PRDM1 as a result of DNA methylation. Overexpression of PRDM1 in EBV-positive BL cell lines resulted in cell cycle arrest. Our results expand the spectrum of lymphoid malignancies in which PRDM1 may have a tumor suppressor role and identify an epigenetic event that likely contributes to the pathogenesis of BL.

  6. Pretreatment dietary intake is associated with tumor suppressor DNA methylation in head and neck squamous cell carcinomas

    Science.gov (United States)

    Colacino, Justin A.; Arthur, Anna E.; Dolinoy, Dana C.; Sartor, Maureen A.; Duffy, Sonia A.; Chepeha, Douglas B.; Bradford, Carol R.; Walline, Heather M.; McHugh, Jonathan B.; D'Silva, Nisha; Carey, Thomas E.; Wolf, Gregory T.; Taylor, Jeremy M.G.; Peterson, Karen E.; Rozek, Laura S.

    2012-01-01

    Diet is associated with cancer prognosis, including head and neck cancer (HNC), and has been hypothesized to influence epigenetic state by determining the availability of functional groups involved in the modification of DNA and histone proteins. The goal of this study was to describe the association between pretreatment diet and HNC tumor DNA methylation. Information on usual pretreatment food and nutrient intake was estimated via food frequency questionnaire (FFQ) on 49 HNC cases. Tumor DNA methylation patterns were assessed using the Illumina Goldengate Methylation Cancer Panel. First, a methylation score, the sum of individual hypermethylated tumor suppressor associated CpG sites, was calculated and associated with dietary intake of micronutrients involved in one-carbon metabolism and antioxidant activity, and food groups abundant in these nutrients. Second, gene specific analyses using linear modeling with empirical Bayesian variance estimation were conducted to identify if methylation at individual CpG sites was associated with diet. All models were controlled for age, sex, smoking, alcohol and HPV status. Individuals reporting in the highest quartile of folate, vitamin B12 and vitamin A intake, compared with those in the lowest quartile, showed significantly less tumor suppressor gene methylation, as did patients reporting the highest cruciferous vegetable intake. Gene specific analyses identified differential associations between DNA methylation and vitamin B12 and vitamin A intake when stratifying by HPV status. These preliminary results suggest that intake of folate, vitamin A and vitamin B12 may be associated with the tumor DNA methylation profile in HNC and enhance tumor suppression. PMID:22722388

  7. Pretreatment with UV light renders the chromatin in human fibroblasts more susceptible to the DNA-damaging agents bleomycin, gamma radiation and 8-methoxypsoralen

    International Nuclear Information System (INIS)

    Ljungman, Mats

    1989-01-01

    Confluent human fibroblast cultures were pretreated with either 254 nm UV light (UV) or methyl methanesulphonate (MMS), incubated at 37 0 C and subsequently challenged on ice with bleomycin (BLM), gamma-radiation or 8-methoxy-psoralen (MOP). The resulting number of challenge-induced DNA damages (measured as DNA strand breaks or cross-links) were compared with the numbers induced in similarly challenged but non-pretreated control cells. It was found that the timing of the subsequent challenge of cells pretreated with UV did significantly affect the amount of induced DNA damage. When the challenging agents were administered after a 10-20 min incubation period following UV pretreatment, the amount of induced DNA damage was increased 50% over control cells. In contrast, the timing of the subsequent challenge of cells pretreated with MMS has no influence on the level of challenge-induced damage. It is hypothesized that UV-irradiated chromatin undergoes a time-dependent decondensation that renders it more susceptible to the induction of strand breaks and cross-links by BLM, gamma-radiation and MOP. A possible role for chromatin decondensation in UV-induced excision repair is discussed. (author)

  8. Epstein-Barr virus (EBV)-encoded small RNAs (EBERs) associated with poor prognosis of head and neck carcinomas.

    Science.gov (United States)

    Turunen, Aaro; Rautava, Jaana; Grénman, Reidar; Syrjänen, Kari; Syrjänen, Stina

    2017-04-18

    Epstein-Barr virus (EBV) is the main cause of nasopharyngeal carcinoma (NPC), also found in other head and neck carcinomas (HNSCCs) where its role remains controversial. EBV was found in 80% and 21% of the samples with PCR and ISH (in cancer cells), respectively. Eight of ISH-positive samples were not NPCs. EBER-RNA detection in carcinoma cells was associated with worse prognosis, whether or not NPCs were included. HPV/EBV and HSV/HPV coinfections associated with a shorter survival. LMP-1 expression, positive in 51% of samples did not correlate with the disease outcome. We analyzed EBV in 73 HNSCC samples with a known HPV and HSV-1 status, using in situ hybridization (ISH) and immunohistochemistry (IHC) for EBV-early transcripts (EBER) and LMP-1 protein, respectively. EBV-DNA was detected with a Luminex-based method. The results were correlated with HPV-status and disease outcome. EBV is transcriptionally active in NPC cells but also in a subgroup of other HNSCCs.

  9. Pathogenic Role of Exosomes in Epstein-Barr Virus (EBV)-Associated Cancers

    Science.gov (United States)

    Teow, Sin-Yeang; Liew, Kitson; Khoo, Alan Soo-Beng; Peh, Suat-Cheng

    2017-01-01

    Exosomes are 40- to 100-nm membrane-bound small vesicles that carry a great variety of cellular cargoes including proteins, DNA, messenger RNAs (mRNAs), and microRNAs (miRNAs). These nanovesicles are detected in various biological fluids such as serum, urine, saliva, and seminal fluids. Exosomes serve as key mediators in intercellular communication by facilitating the transfer and exchange of cellular components from cells to cells. They contain various pathogenic factors whereby their adverse effects have been implicated in multiple viral infections and cancers. Interestingly, accumulating evidences showed that exosomes derived from tumour viruses or oncoviruses, exacerbate virus-associated cancers by remodelling the tumour microenvironment. In this review, we summarize the contributing factors of Epstein-Barr virus (EBV) products-containing exosomes in viral pathogenesis and their potential implications in EBV-driven malignancies. Understanding the biological role of these exosomes in the disease would undoubtedly boost the development of a more comprehensive strategy to combat EBV-associated cancers and to better predict the therapeutic outcomes. Furthermore, we also highlight the potentials and challenges of EBV products-containing exosomes being employed as diagnostic markers and therapeutic targets for EBV-related cancers. Since these aspects are rather underexplored, we attempt to underline interesting areas that warrant further investigations in the future. PMID:29104494

  10. Antibodies to early EBV, CMV, and HHV6 antigens in systemic lupus erythematosus patients

    DEFF Research Database (Denmark)

    Rasmussen, N S; Draborg, A H; Nielsen, C T

    2015-01-01

    /biochemistry, serology, and disease activity measures. METHOD: Immunoglobulin (Ig)M, IgG, and IgA levels against the DNA polymerase processivity factors of EBV, CMV, and HHV6, termed early antigen diffuse (EA/D), pp52, and p41, respectively, were determined in plasma samples from 77 SLE patients and 29 healthy controls...... by using enzyme-linked immunosorbent assays (ELISAs). RESULTS: IgM, IgG, and IgA levels against EBV EA/D, and IgG and IgA levels against CMV pp52, were significantly higher in SLE patients compared with healthy controls. Furthermore, EBV EA/D- and CMV pp52-directed IgG levels were inversely and positively...... associated, respectively, with lymphocyte counts in SLE patients. None of the findings seemed to be associated with use of immunosuppressive medication. CONCLUSIONS: Our results suggest strong, but opposite, associations of lytic EBV and CMV infections with SLE. The amplified humoral responses to EBV EA/D...

  11. Pathogenic Role of Exosomes in Epstein-Barr Virus (EBV)-Associated Cancers.

    Science.gov (United States)

    Teow, Sin-Yeang; Liew, Kitson; Khoo, Alan Soo-Beng; Peh, Suat-Cheng

    2017-01-01

    Exosomes are 40- to 100-nm membrane-bound small vesicles that carry a great variety of cellular cargoes including proteins, DNA, messenger RNAs (mRNAs), and microRNAs (miRNAs). These nanovesicles are detected in various biological fluids such as serum, urine, saliva, and seminal fluids. Exosomes serve as key mediators in intercellular communication by facilitating the transfer and exchange of cellular components from cells to cells. They contain various pathogenic factors whereby their adverse effects have been implicated in multiple viral infections and cancers. Interestingly, accumulating evidences showed that exosomes derived from tumour viruses or oncoviruses, exacerbate virus-associated cancers by remodelling the tumour microenvironment. In this review, we summarize the contributing factors of Epstein-Barr virus (EBV) products-containing exosomes in viral pathogenesis and their potential implications in EBV-driven malignancies. Understanding the biological role of these exosomes in the disease would undoubtedly boost the development of a more comprehensive strategy to combat EBV-associated cancers and to better predict the therapeutic outcomes. Furthermore, we also highlight the potentials and challenges of EBV products-containing exosomes being employed as diagnostic markers and therapeutic targets for EBV-related cancers. Since these aspects are rather underexplored, we attempt to underline interesting areas that warrant further investigations in the future.

  12. EBV Latency Types Adopt Alternative Chromatin Conformations

    Science.gov (United States)

    Tempera, Italo; Klichinsky, Michael; Lieberman, Paul M.

    2011-01-01

    Epstein-Barr Virus (EBV) can establish latent infections with distinct gene expression patterns referred to as latency types. These different latency types are epigenetically stable and correspond to different promoter utilization. Here we explore the three-dimensional conformations of the EBV genome in different latency types. We employed Chromosome Conformation Capture (3C) assay to investigate chromatin loop formation between the OriP enhancer and the promoters that determine type I (Qp) or type III (Cp) gene expression. We show that OriP is in close physical proximity to Qp in type I latency, and to Cp in type III latency. The cellular chromatin insulator and boundary factor CTCF was implicated in EBV chromatin loop formation. Combining 3C and ChIP assays we found that CTCF is physically associated with OriP-Qp loop formation in type I and OriP-Cp loop formation in type III latency. Mutations in the CTCF binding site located at Qp disrupt loop formation between Qp and OriP, and lead to the activation of Cp transcription. Mutation of the CTCF binding site at Cp, as well as siRNA depletion of CTCF eliminates both OriP-associated loops, indicating that CTCF plays an integral role in loop formation. These data indicate that epigenetically stable EBV latency types adopt distinct chromatin architectures that depend on CTCF and mediate alternative promoter targeting by the OriP enhancer. PMID:21829357

  13. EBV latency types adopt alternative chromatin conformations.

    Directory of Open Access Journals (Sweden)

    Italo Tempera

    2011-07-01

    Full Text Available Epstein-Barr Virus (EBV can establish latent infections with distinct gene expression patterns referred to as latency types. These different latency types are epigenetically stable and correspond to different promoter utilization. Here we explore the three-dimensional conformations of the EBV genome in different latency types. We employed Chromosome Conformation Capture (3C assay to investigate chromatin loop formation between the OriP enhancer and the promoters that determine type I (Qp or type III (Cp gene expression. We show that OriP is in close physical proximity to Qp in type I latency, and to Cp in type III latency. The cellular chromatin insulator and boundary factor CTCF was implicated in EBV chromatin loop formation. Combining 3C and ChIP assays we found that CTCF is physically associated with OriP-Qp loop formation in type I and OriP-Cp loop formation in type III latency. Mutations in the CTCF binding site located at Qp disrupt loop formation between Qp and OriP, and lead to the activation of Cp transcription. Mutation of the CTCF binding site at Cp, as well as siRNA depletion of CTCF eliminates both OriP-associated loops, indicating that CTCF plays an integral role in loop formation. These data indicate that epigenetically stable EBV latency types adopt distinct chromatin architectures that depend on CTCF and mediate alternative promoter targeting by the OriP enhancer.

  14. et d'Epstein-Barr virus (EBV)

    African Journals Online (AJOL)

    Diagnostic moléculaire du Cytomégalovirus (CMV), de l'herpès virus humain de type 6 (HHV6) et d'Epstein-Barr virus (EBV) par PCR en temps réel chez les femmes enceintes VIH séropositives et séronégatives à Ouagadougou, Burkina Faso.

  15. Genipin as a novel chemical activator of EBV lytic cycle.

    Science.gov (United States)

    Son, Myoungki; Lee, Minjung; Ryu, Eunhyun; Moon, Aree; Jeong, Choon-Sik; Jung, Yong Woo; Park, Gyu Hwan; Sung, Gi-Ho; Cho, Hyosun; Kang, Hyojeung

    2015-02-01

    Epstein-Barr virus (EBV) is a ubiquitous gammaherpesvirus that causes acute infection and establishes life-long latency. EBV causes several human cancers, including Burkitt's lymphoma, nasopharyngeal and gastric carcinoma. Antiviral agents can be categorized as virucides, antiviral chemotherapeutic agents, and immunomodulators. Most antiviral agents affect actively replicating viruses, but not their latent forms. Novel antiviral agents must be active on both the replicating and the latent forms of the virus. Gardenia jasminoides is an evergreen flowering plant belonging to the Rubiaceae family and is most commonly found growing wild in Vietnam, Southern China, Taiwan, Japan, Myanmar, and India. Genipin is an aglycone derived from an iridoid glycoside called geniposide, which is present in large quantities in the fruit of G. jasminoides. In this study, genipin was evaluated for its role as an antitumor and antiviral agent that produces inhibitory effects against EBV and EBV associated gastric carcinoma (EBVaGC). In SNU719 cells, one of EBVaGCs, genipin caused significant cytotoxicity (70 μM), induced methylation on EBV C promoter and tumor suppressor gene BCL7A, arrested cell-cycle progress (S phases), upregulated EBV latent/lytic genes in a dose-dependent manner, stimulated EBV progeny production, activated EBV F promoter for EBV lytic activation, and suppressed EBV infection. These results indicated that genipin could be a promising candidate for antiviral and antitumor agents against EBV and EBVaGC.

  16. Prevalence and molecular profiling of Epstein Barr virus (EBV among healthy blood donors from different nationalities in Qatar.

    Directory of Open Access Journals (Sweden)

    Maria K Smatti

    Full Text Available The Epstein-Barr virus (EBV is the causative agent of infectious mononucleosis. EBV is highly prevalent lymphotropic herpesvirus and has been linked to several malignancies. Transmission is generally by oral secretions, but can be through blood transfusions and organ transplantations. This study aimed to determine the seroprevalence, viremia rates, and circulating genotypes of EBV in healthy blood donors in Qatar.Blood samples from 673 blood donors of different nationalities residing in Qatar (mainly Qatar, Egypt, Syria, Jordan, Pakistan, and India were collected and tested for anti-EBV capsid (VCA; IgG & IgM, nuclear (EBNA; IgG, and early (EA-D; IgG antigens. Avidity testing was determined when active infection was suspected. DNA was extracted from the buffy coat and subjected to EBV-DNA quantification using qRT-PCR. Genotyping was performed using nested-PCR targeting EBV-EBNA2 gene, and phylogeny by sequence analysis of the LMP-1 gene.97.9% (673/659 of the samples were seropositive as indicated by the presence VCA-IgG, while 52.6% (354/673 had detectible EBV-DNA. EBV seroprevalence and viremia rates increased significantly with age. Genotyping of 51 randomly selected samples showed predominance of Genotype 1 (72.5%, 37/51 as compared to genotype 2 (3.5%, and mixed infections were detected in 4% of the samples. Sub-genotyping for these samples revealed that the Mediterranean strain was predominant (65.3%, followed by B95.8 prototype and North Carolina strains (12.2% each, and China1 strain (6%.As a first study to evaluate EBV infection in highly diverse population in Qatar, where expatriates represent more than 85% of the population, our results indicated high seroprevalence and viremia rate of EBV in different nationalities, with genotype 1 and Mediterranean strain being predominant. Clinical significance of these finding have not been investigated and shall be evaluated in future studies.

  17. Endoplasmic reticulum stress causes EBV lytic replication

    OpenAIRE

    Taylor, Gwen Marie; Raghuwanshi, Sandeep K.; Rowe, David T.; Wadowsky, Robert M.; Rosendorff, Adam

    2011-01-01

    Endoplasmic reticulum (ER) stress triggers a homeostatic cellular response in mammalian cells to ensure efficient folding, sorting, and processing of client proteins. In lytic-permissive lymphoblastoid cell lines (LCLs), pulse exposure to the chemical ER-stress inducer thapsigargin (TG) followed by recovery resulted in the activation of the EBV immediate-early (BRLF1, BZLF1), early (BMRF1), and late (gp350) genes, gp350 surface expression, and virus release. The protein phosphatase 1 a (PP1a)...

  18. Frequency and clinical correlates of elevated plasma Epstein-Barr virus DNA at diagnosis in peripheral T-cell lymphomas

    Science.gov (United States)

    Haverkos, Bradley M.; Huang, Ying; Gru, Alejandro; Pancholi, Preeti; Freud, Aharon G.; Mishra, Anjali; Ruppert, Amy S.; Baiocchi, Robert A.; Porcu, Pierluigi

    2016-01-01

    Epstein Barr virus (EBV)-encoded RNAs (EBER) in tumor tissue and cell-free plasma EBV-DNA (pEBVd) are detected in EBV-associated lymphomas. Studies have suggested that EBER+ peripheral T-cell lymphomas (PTCL) have worse prognosis, but the role of EBV in these neoplasms remains unclear. pEBVd is quantitative and more easily amenable to standardization than EBER, but frequency of pEBVd detection, clinical impact, and agreement with EBER status in PTCL are unknown. We retrospectively assessed frequency of detectable pre-treatment pEBVd, presence of EBER in tumor tissue, and outcomes in 61 of 135 EBV-assessable PTCL patients. Fifteen of 61 patients (24.5%, 95% CI: 14–37%) were pre-treatment pEBVd+, with no significant differences in baseline characteristics or treatment between pEBVd+ and pEBVd− patients. EBER-ISH was performed on 10 pEBVd+ and 35 pEBVd− tumors. All 10 pEBVd+ patients were EBER+, but 9 pEBVd− patients were also EBER+. With median follow up of 24 months (range 1–96), overall survival (OS) was shorter in pEBVd+ compared to pEBVd− patients (13 vs. 72 months; p=0.04). In this retrospective study, pre-treatment pEBVd was elevated in 25% of PTCL patients, was highly specific for EBER+ tumors, and was associated with shorter survival. pEBVd should be further explored as a prognostic variable and tumor biomarker in PTCL. PMID:27943278

  19. Endoplasmic reticulum stress causes EBV lytic replication.

    Science.gov (United States)

    Taylor, Gwen Marie; Raghuwanshi, Sandeep K; Rowe, David T; Wadowsky, Robert M; Rosendorff, Adam

    2011-11-17

    Endoplasmic reticulum (ER) stress triggers a homeostatic cellular response in mammalian cells to ensure efficient folding, sorting, and processing of client proteins. In lytic-permissive lymphoblastoid cell lines (LCLs), pulse exposure to the chemical ER-stress inducer thapsigargin (TG) followed by recovery resulted in the activation of the EBV immediate-early (BRLF1, BZLF1), early (BMRF1), and late (gp350) genes, gp350 surface expression, and virus release. The protein phosphatase 1 a (PP1a)-specific phosphatase inhibitor Salubrinal (SAL) synergized with TG to induce EBV lytic genes; however, TG treatment alone was sufficient to activate EBV lytic replication. SAL showed ER-stress-dependent and -independent antiviral effects, preventing virus release in human LCLs and abrogating gp350 expression in 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated B95-8 cells. TG resulted in sustained BCL6 but not BLIMP1 or CD138 expression, which is consistent with maintenance of a germinal center B-cell, rather than plasma-cell, phenotype. Microarray analysis identified candidate genes governing lytic replication in LCLs undergoing ER stress.

  20. Prospective study of tailoring whole-body dual-modality [18F]fluorodeoxyglucose positron emission tomography/computed tomography with plasma Epstein-Barr virus DNA for detecting distant metastasis in endemic nasopharyngeal carcinoma at initial staging.

    Science.gov (United States)

    Tang, Lin-Quan; Chen, Qiu-Yan; Fan, Wei; Liu, Huai; Zhang, Lu; Guo, Ling; Luo, Dong-Hua; Huang, Pei-Yu; Zhang, Xu; Lin, Xiao-Ping; Mo, Yun-Xian; Liu, Li-Zhi; Mo, Hao-Yuan; Li, Jian; Zou, Ru-Hai; Cao, Yun; Xiang, Yan-Qun; Qiu, Fang; Sun, Rui; Chen, Ming-Yuan; Hua, Yi-Jun; Lv, Xing; Wang, Lin; Zhao, Chong; Guo, Xiang; Cao, Ka-Jia; Qian, Chao-Nan; Zeng, Mu-Sheng; Mai, Hai-Qiang

    2013-08-10

    To evaluate which patients with nasopharyngeal carcinoma (NPC) obtained the greatest benefits from the detection of distant metastasis with [(18)F]fluorodeoxyglucose positron emission tomography and computed tomography (PET/CT) combined with plasma Epstein-Barr virus (EBV) DNA levels. Consecutive patients with NPC were prospectively enrolled. PET/CT, conventional work-up (CWU), and quantification of plasma EBV DNA were performed before treatment. The accuracy of these strategies for distant metastases was assessed. The costs of the diagnostic strategies were compared. Eighty-six (14.8%) of the 583 eligible patients were found to have distant metastases; 71 patients (82.6%) by PET/CT and 31 patients (36.0%) by CWU. In the multivariable analysis, advanced N stage (odds ratio, 2.689; 95% CI, 1.894 to 3.818) and pretreatment EBV DNA level (odds ratio, 3.344; 95% CI, 1.825 to 6.126) were significant risk factors for distant metastases. PET/CT was not superior to CWU for detecting distant metastases in very low-risk patients (N0-1 with EBV DNA < 4,000 copies/mL; P = .062), but was superior for the low-risk patients (N0-1 with EBV DNA ≥ 4,000 copies/mL and N2-3 with EBV DNA < 4,000 copies/mL; P = .039) and intermediate-risk patients (N2-3 disease with EBV DNA ≥ 4,000 copies/mL; P < .001). The corresponding patient management changes based on PET/CT were 2.9%, 6.3%, and 16.5%, respectively. The costs per true-positive case detected by PET/CT among these groups were ¥324,138 (≈$47,458), ¥96,907 (≈$14,188), and ¥34,182 (≈$5,005), respectively. PET/CT detects more distant metastases than conventional staging in patients with NPC. The largest benefit in terms of cost and patient management was observed in the subgroup with N2-3 disease and EBV DNA ≥ 4,000 copies/mL.

  1. Occurrence of antibiotic resistance genes and mobile genetic elements in enterococci and genomic DNA during anaerobic digestion of pharmaceutical waste sludge with different pretreatments.

    Science.gov (United States)

    Tong, Juan; Lu, XueTing; Zhang, JunYa; Sui, Qianwen; Wang, Rui; Chen, Meixue; Wei, Yuansong

    2017-07-01

    Pharmaceutical waste sludge harbors large amounts of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs), and it is necessary to study the reduction of ARGs and MGEs during sludge treatment. Therefore, the antibiotic resistance phenotypes and genotypes of enterococci, and the ARGs and MGEs in genomic DNA were investigated during anaerobic digestion (AD) with microwave (MW), thermal hydrolysis (TH) and ozone pretreatment. Results showed that sludge pretreatment increased the occurrence of the resistance phenotypes and genotypes of enterococci. During AD, the resistance of enterococci to macrolides decreased, except for in the MW-pretreated sludge. Horizontal gene transfer and co-occurrence of ermB and tetM in enterococci resulted in increased tetracycline resistance of enterococci throughout the sludge treatment. MGEs such as intI1, ISCR1 and Tn916/1545 had a significant effect on the distribution of ARGs. AD with pretreatment, especially TH pretreatment, resulted in greater ARGs and MGEs reduction and improved methane production. Copyright © 2017. Published by Elsevier Ltd.

  2. Prevalence of HPV and EBV infection and their relationship with the p53 and PCNA expression in oral carcinoma patients.

    Directory of Open Access Journals (Sweden)

    Dayahindara Veitía

    2017-04-01

    Full Text Available Introduction: Infection caused by potentially oncogenic viruses, such as HPV and EBV, favors the role of certain oncoproteins that can induce dysplasias and malignant lesions. Objective: To evaluate the prevalence of HPV and EBV and their relation with the expression of p53 and PCNA in patients with oral carcinoma. Methodology: Twenty-seven oral squamous cell carcinomas (OSCC were evaluated; DNA extraction was conducted using the QIAamp DNA mini kit; viral detection was obtained using the INNO-LiPA kit for HPV, and nested PCR was used for EBV. The evaluation of molecular markers was performed through immunohistochemical staining. Results: The mean age of the patients was 60.55±13.94 years, and 52% of these were female. Of the patients, 59% were tobacco users and 63% were alcohol consumers. HPV was detected in 70% of the patients with the predominance of genotype 16 (60%. As for EBV infection, it was observed in 59% of cases. p53 and PCNA immunopositivity corresponded to 44% and 59%, respectively. The tongue was the anatomical location with highest positivity for both viruses as well as for the expression of molecular markers. The 48% of the cases presented infection by both viruses. Conclusion: HPV and EBV infection together with the expression of p53 and PCNA were more frequently observed in advanced stages of the disease, suggesting a more relevant role in the progression than in tumor genesis.

  3. Epstein-Barr Virus (EBV)-associated Gastric Carcinoma

    Science.gov (United States)

    Iizasa, Hisashi; Nanbo, Asuka; Nishikawa, Jun; Jinushi, Masahisa; Yoshiyama, Hironori

    2012-01-01

    The ubiquitous Epstein-Barr virus (EBV) is associated with several human tumors, which include lymphoid and epithelial malignancies. It is known that EBV persistently infects the memory B cell pool of healthy individuals by activating growth and survival signaling pathways that can contribute to B cell lymphomagenesis. Although the monoclonal proliferation of EBV-infected cells can be observed in epithelial tumors, such as nasopharyngeal carcinoma and EBV-associated gastric carcinoma, the precise role of EBV in the carcinogenic progress is not fully understood. This review features characteristics and current understanding of EBV-associated gastric carcinoma. EBV-associated gastric carcinoma comprises almost 10% of all gastric carcinoma cases and expresses restricted EBV latent genes (Latency I). Firstly, definition, epidemiology, and clinical features are discussed. Then, the route of infection and carcinogenic role of viral genes are presented. Of particular interest, the association with frequent genomic CpG methylation and role of miRNA for carcinogenesis are topically discussed. Finally, the possibility of therapies targeting EBV-associated gastric carcinoma is proposed. PMID:23342366

  4. BZLF1 Expression of EBV is correlated with PARP1 Regulation on Nasopharyngeal Carcinoma Tissues

    Directory of Open Access Journals (Sweden)

    Wahyu nur laili fajri, Ahmad Rofi'i, Fatchiyah Fatchiyah

    2013-04-01

    , but not correlated with percentage of necrotic cell. Interestingly, in all tissues (NPC-T1, NPC-T2 and polyp is increased number of karyolysis cells properly, but not in pyknosis and karryorrhexis cells. The conclusion of this study indicated that the BZLF1 induce PARP1 to repair DNA damage against EBV infection.

  5. Evaluation of the presence of Epstein-Barr virus (EBV) in Iranian patients with thyroid papillary carcinoma.

    Science.gov (United States)

    Homayouni, Maryam; Mohammad Arabzadeh, Seyed Ali; Nili, Fatemeh; Razi, Farideh; Amoli, Mahsa Mohammad

    2017-07-01

    Papillary thyroid carcinoma (PTC) is the most common thyroid cancer. EBV is one of the most important viruses related to different types of malignancies. This study investigated the relationship between EBV and papillary thyroid carcinoma. In this study the presence of Epstein-Barr Nuclear Antigen 1 (EBNA1) gene in papillary thyroid carcinoma tissues were examined by nested-PCR method. Paraffin-embedded tissues (N=41) blocks of thyroid cancer were used. DNA was extracted from all samples and then samples were evaluated for the presence of EBV gene. In 41 samples, EBNA1 was detected in 65.8% of patients with papillary thyroid carcinoma which was significantly higher in younger ages. The significant presence of EBV genome in papillary thyroid carcinoma suggests that this virus may play a role in this cancer especially in younger ages. As a result, monitoring of patients with EBV latent infection for PTC can be very important. Copyright © 2017 Elsevier GmbH. All rights reserved.

  6. A novel approach to detect KRAS/BRAF mutation for colon cancer: Highly sensitive simultaneous detection of mutations and simple pre-treatment without DNA extraction.

    Science.gov (United States)

    Suzuki, Shun-Ichi; Matsusaka, Satoshi; Hirai, Mitsuharu; Shibata, Harumi; Takagi, Koichi; Mizunuma, Nobuyuki; Hatake, Kiyohiko

    2015-07-01

    It has been reported that colon cancer patients with KRAS and BRAF mutations that lie downstream of epidermal growth factor receptor (EGFR) acquire resistance against therapy with anti‑EGFR antibodies, cetuximab and panitumumab. On the other hand, some reports say KRAS codon 13 mutation (p.G13D) has lower resistance against anti-EGFR antibodies, thus there is a substantial need for detection of specific KRAS mutations. We have established a state-of-the-art measurement system using QProbe (QP) method that allows simultaneous measurement of KRAS codon 12/13, p.G13D and BRAF mutation, and compared this method against Direct Sequencing (DS) using 182 specimens from colon cancer patients. In addition, 32 biopsy specimens were processed with a novel pre-treatment method without DNA purification in order to detect KRAS/BRAF. As a result of KRAS mutation measurement, concordance rate between the QP method and DS method was 81.4% (144/177) except for the 5 specimens that were undeterminable. Among them, 29 specimens became positive with QP method and negative with DS method. BRAF was measured with QP method only, and the mutation detection rate was 3.9% (6/153). KRAS measurement using a simple new pre-treatment method without DNA extraction resulted in 31 good results out of 32, all of them matching with the DS method. We have established a simple but highly sensitive simultaneous detection system for KRAS/BRAF. Moreover, introduction of the novel pre-treatment technology eliminated the inconvenient DNA extraction process. From this research achievement, we not only anticipate quick and accurate results returned in the clinical field but also contribution in improving the test quality and work efficiency.

  7. Physical Pre-Treatment Improves Efficient DNA Extraction and qPCR Sensitivity from Clostridium Difficile Spores in Faecal Swine Specimens.

    Science.gov (United States)

    Grześkowiak, Łukasz; Zentek, Jürgen; Vahjen, Wilfried

    2016-11-01

    A considerable fraction of the faecal microbiota is spore-forming. Molecular quantification of bacteria may be underestimated if preceded with nucleic acid extraction without special treatment to extract recalcitrant bacterial spores. The objective of this study was to improve the DNA extraction regarding the presence of Clostridium difficile spores in faecal swine specimens. Sow faeces were inoculated with spores of C. difficile (10(6) CFU), frozen at - 30 °C overnight and subjected to DNA extraction. As a preceding step to a standard DNA extraction method (QIAamp DNA stool Mini kit), different physical treatments such as microwave oven heating and repeated bead-beating techniques and a combination of both were applied and compared with each other by means of qPCR. Using a standard DNA extraction method only, C. difficile spores were quantified at 4.96 log copy number/200 mg of faeces. A repeated bead-beating at 6 m/s for 10 min followed by a standard DNA extraction resulted in 5.77 log copy number of spores in inoculated faeces. Heating in a microwave oven at 800 W for 1, 3, 5 and 10 min followed by a standard DNA extraction resulted in a gene quantification of up to 4.89 log copy number. A combination of both methods resulted in the bacterial gene quantity of 5.37 log copy number. Pre-treatment with repeated bead-beating led to the highest quantification of bacteria, and therefore it can be applied for more efficient DNA extraction from spores of C. difficile in faecal specimens.

  8. Transplantation of solid organ recipients shedding Epstein-Barr virus DNA pre-transplant: A prospective study.

    Science.gov (United States)

    Verghese, Priya S; Schmeling, David O; Filtz, Emma A; Grimm, Jennifer M; Matas, Arthur J; Balfour, Henry H

    2017-11-01

    Epstein-Barr virus (EBV) poses a significant threat to patient and graft survival post-transplant. We hypothesized that recipients who shed EBV at transplant had less immunologic control of the virus and hence were more likely to have active EBV infection and disease post-transplant. To test this hypothesis, we conducted a 5-year prospective study in primary solid organ transplant recipients. We measured EBV DNA in oral washes and blood samples by quantitative PCR before transplant and periodically thereafter for up to 4 years. Pre-transplant samples were available from 98 subjects. EBV DNA was detected pre-transplant in 32 of 95 (34%) and 5 of 93 subjects (5%) in oral wash and blood, respectively. Recipients with and without detectable pre-transplant EBV DNA were not significantly different demographically and had no significant difference in patient and graft survival (P = .6 for both comparisons) or post-transplant EBV viremia-free survival (P = .8). There were no cases of EBV-related disease or post-transplant lymphoproliferative disorder (PTLD) in any of the patients with detectable EBV DNA pre-transplant. In conclusion, detectable EBV DNA pre-transplant was not associated with differences in patient/graft survival, post-transplant EBV viremia, or EBV-related diseases including PTLD. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. Individuazione di un nuovo marker da impiegare per una corretta stadiazione dell’infezione da EBV

    Directory of Open Access Journals (Sweden)

    Luana Coltella

    2006-06-01

    Full Text Available Epstein Barr Virus (EBV, also classified as Human Herpes Virus 4, infects the vast majority of adults worldwide and establishes both non-productive (latent and productive (lytic infection. Classical EBV diagnosis includes quantitative determination of viral DNA and serological analysis, based on the determination of IgG and IgM responses against the viral capsid antigen (VCA and the IgG response against the EBV nuclear antigen-1 (EBNA-1. EBV-serology can be misleading in some cases, such as acute infections with low or undetectable VCA IgM, convalescent patients with persistent or reactivated VCA IgM and negative anti- EBNA-1 IgG, due to a loss of this marker during immunosuppression. In all these cases avidity determination of IgG is helpful to prevent false diagnosis. Avidity represents the stability of the antigen-antibody interaction. Its value increases during the infection, so high avidity never associates with a primary acute infection. We studied the importance of avidity determination of p18 (VCA-IgG to achieve unequivocal interpretation of serological results. The amount of IgG and IgM is determined by Chemiluminescent Immune Assay (CLIA, a rapid and highly sensitive method suitable for automation. The intensity of luminous signal produced by antibody-antigen recognition is expressed as Relative Light Unit (RLU. p-18 IgG is determined using a recombinant p18 antigen expressed in E. coli. Avidity index is determined in CLIA by the ratio between denaturated and not denaturated IgG specific antibodies expressed in RLU. These results demonstrate that avidity determination represents an important additional marker particularly in cases with aberrant serological profile.

  10. EBV-Associated Cancer and Autoimmunity: Searching for Therapies

    Directory of Open Access Journals (Sweden)

    Giovanni Capone

    2015-02-01

    Full Text Available Epstein-Barr virus (EBV infects B-, T-, and NK cells and has been associated not only with a wide range of lymphoid malignancies but also with autoimmune diseases such as lupus erythematosus, rheumatoid arthritis and, in particular, multiple sclerosis. Hence, effective immunotherapeutic approaches to eradicate EBV infection might overthrow cancer and autoimmunity incidence. However, currently no effective anti-EBV immunotherapy is available. Here we use the concept that protein immunogenicity is allocated in rare peptide sequences and search the Epstein-Barr nuclear antigen 1 (EBNA1 sequence for peptides unique to the viral protein and absent in the human host. We report on a set of unique EBV EBNA1 peptides that might be used in designing peptide-based therapies able to specifically hitting the virus or neutralizing pathogenic autoantibodies.

  11. Pivotal role of HIV and EBV replication in the long-term persistence of monoclonal gammopathy in patients on antiretroviral therapy.

    Science.gov (United States)

    Ouedraogo, David Eric; Makinson, Alain; Vendrell, Jean-Pierre; Casanova, Marie-Laure; Nagot, Nicolas; Cezar, Renaud; Bollore, Karine; Al Taaba, Yassine; Foulongne, Vincent; Badiou, Stéphanie; Viljoen, Johannes; Reynes, Jacques; Van de Perre, Philippe; Tuaillon, Edouard

    2013-10-24

    A high prevalence of monoclonal gammopathy (MG) has been observed in HIV-infected patients. We explored the conditions associated with long-term persistence of serum monoclonal protein (M protein) in HIV-infected patients on antiretroviral therapy (ART). Of 21 patients with MG, M protein disappeared in 12 patients (58%) over 5 years of ART. Higher level of serum γ-globulin and higher percentages of circulating plasmablasts and plasma cells were observed in patients with persistent MG compared with patients with transient MG. MG persistence was associated with the cumulative time of detectable plasma HIV RNA after ART initiation, detection of Epstein-Barr virus (EBV) DNA in plasma, and a high level of EBV DNA in B cells. Poor control of HIV replication and detectable EBV replication in plasma were both associated with long-term MG persistence in patients on ART. In the case of viral control, MG associated with HIV infection is usually transient.

  12. Biomass pretreatment

    Science.gov (United States)

    Hennessey, Susan Marie; Friend, Julie; Elander, Richard T; Tucker, III, Melvin P

    2013-05-21

    A method is provided for producing an improved pretreated biomass product for use in saccharification followed by fermentation to produce a target chemical that includes removal of saccharification and or fermentation inhibitors from the pretreated biomass product. Specifically, the pretreated biomass product derived from using the present method has fewer inhibitors of saccharification and/or fermentation without a loss in sugar content.

  13. Epstein-Barr virus (EBV) provides survival factors to EBV+diffuse large B-cell lymphoma (DLBCL) lines and modulates cytokine induced specific chemotaxis in EBV+ DLBCL.

    Science.gov (United States)

    Wu, Liang; Ehlin-Henriksson, Barbro; Zhou, Xiaoying; Zhu, Hong; Ernberg, Ingemar; Kis, Lorand L; Klein, George

    2017-12-01

    Diffuse large B-cell lymphoma (DLBCL), the most common type of malignant lymphoma, accounts for 30% of adult non-Hodgkin lymphomas. Epstein-Barr virus (EBV) -positive DLBCL of the elderly is a newly recognized subtype that accounts for 8-10% of DLBCLs in Asian countries, but is less common in Western populations. Five DLBCL-derived cell lines were employed to characterize patterns of EBV latent gene expression, as well as response to cytokines and chemotaxis. Interleukin-4 and interleukin-21 modified LMP1, EBNA1 and EBNA2 expression depending on cell phenotype and type of EBV latent programme (type I, II or III). These cytokines also affected CXCR4- or CCR7-mediated chemotaxis in two of the cell lines, Farage (type III) and Val (type II). Further, we investigated the effect of EBV by using dominant-negative EBV nuclear antigen 1(dnEBNA1) to eliminate EBV genomes. This resulted in decreased chemotaxis. By employing an alternative way to eliminate EBV genomes, Roscovitine, we show an increase of apoptosis in the EBV-positive lines. These results show that EBV plays an important role in EBV-positive DLBCL lines with regard to survival and chemotactic response. Our findings provide evidence for the impact of microenvironment on EBV-carrying DLBCL cells and might have therapeutic implications. © 2017 John Wiley & Sons Ltd.

  14. Epstein-Barr virus (EBV Rta-mediated EBV and Kaposi's sarcoma-associated herpesvirus lytic reactivations in 293 cells.

    Directory of Open Access Journals (Sweden)

    Yen-Ju Chen

    Full Text Available Epstein-Barr virus (EBV Rta belongs to a lytic switch gene family that is evolutionarily conserved in all gamma-herpesviruses. Emerging evidence indicates that cell cycle arrest is a common means by which herpesviral immediate-early protein hijacks the host cell to advance the virus's lytic cycle progression. To examine the role of Rta in cell cycle regulation, we recently established a doxycycline (Dox-inducible Rta system in 293 cells. In this cell background, inducible Rta modulated the levels of signature G1 arrest proteins, followed by induction of the cellular senescence marker, SA-β-Gal. To delineate the relationship between Rta-induced cell growth arrest and EBV reactivation, recombinant viral genomes were transferred into Rta-inducible 293 cells. Somewhat unexpectedly, we found that Dox-inducible Rta reactivated both EBV and Kaposi's sarcoma-associated herpesvirus (KSHV, to similar efficacy. As a consequence, the Rta-mediated EBV and KSHV lytic replication systems, designated as EREV8 and ERKV, respectively, were homogenous, robust, and concurrent with cell death likely due to permissive lytic replication. In addition, the expression kinetics of EBV lytic genes in Dox-treated EREV8 cells was similar to that of their KSHV counterparts in Dox-induced ERKV cells, suggesting that a common pathway is used to disrupt viral latency in both cell systems. When the time course was compared, cell cycle arrest was achieved between 6 and 48 h, EBV or KSHV reactivation was initiated abruptly at 48 h, and the cellular senescence marker was not detected until 120 h after Dox treatment. These results lead us to hypothesize that in 293 cells, Rta-induced G1 cell cycle arrest could provide (1 an ideal environment for virus reactivation if EBV or KSHV coexists and (2 a preparatory milieu for cell senescence if no viral genome is available. The latter is hypothetical in a transient-lytic situation.

  15. Epstein-Barr Virus Hijacks DNA Damage Response Transducers to Orchestrate Its Life Cycle.

    Science.gov (United States)

    Hau, Pok Man; Tsao, Sai Wah

    2017-11-16

    The Epstein-Barr virus (EBV) is a ubiquitous virus that infects most of the human population. EBV infection is associated with multiple human cancers, including Burkitt's lymphoma, Hodgkin's lymphoma, a subset of gastric carcinomas, and almost all undifferentiated non-keratinizing nasopharyngeal carcinoma. Intensive research has shown that EBV triggers a DNA damage response (DDR) during primary infection and lytic reactivation. The EBV-encoded viral proteins have been implicated in deregulating the DDR signaling pathways. The consequences of DDR inactivation lead to genomic instability and promote cellular transformation. This review summarizes the current understanding of the relationship between EBV infection and the DDR transducers, including ATM (ataxia telangiectasia mutated), ATR (ATM and Rad3-related), and DNA-PK (DNA-dependent protein kinase), and discusses how EBV manipulates the DDR signaling pathways to complete the replication process of viral DNA during lytic reactivation.

  16. DNA Ploidy Measured on Archived Pretreatment Biopsy Material May Correlate With Prostate-Specific Antigen Recurrence After Prostate Brachytherapy

    Energy Technology Data Exchange (ETDEWEB)

    Keyes, Mira, E-mail: mkeyes@bccancer.bc.ca [Radiation Oncology, Provincial Prostate Brachytherapy Program, Vancouver Cancer Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada); MacAulay, Calum [Department of Integrative Oncology, British Columbia Cancer Research Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada); Hayes, Malcolm [Department of Pathology, Vancouver Cancer Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada); Korbelik, Jagoda [Department of Integrative Oncology, British Columbia Cancer Research Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada); Morris, W. James [Radiation Oncology, Provincial Prostate Brachytherapy Program, Vancouver Cancer Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada); Palcic, Branko [Department of Integrative Oncology, British Columbia Cancer Research Centre, British Columbia Cancer Agency, Vancouver, British Columbia (Canada)

    2013-08-01

    Purpose: To explore whether DNA ploidy of prostate cancer cells determined from archived transrectal ultrasound-guided biopsy specimens correlates with disease-free survival. Methods and Materials: Forty-seven failures and 47 controls were selected from 1006 consecutive low- and intermediate-risk patients treated with prostate {sup 125}I brachytherapy (July 1998-October 2003). Median follow-up was 7.5 years. Ten-year actuarial disease-free survival was 94.1%. Controls were matched using age, initial prostate-specific antigen level, clinical stage, Gleason score, use of hormone therapy, and follow-up (all P nonsignificant). Seventy-eight specimens were successfully processed; 27 control and 20 failure specimens contained more than 100 tumor cells were used for the final analysis. The Feulgen-Thionin stained cytology samples from archived paraffin blocks were used to determine the DNA ploidy of each tumor by measuring integrated optical densities. Results: The samples were divided into diploid and aneuploid tumors. Aneuploid tumors were found in 16 of 20 of the failures (80%) and 8 of 27 controls (30%). Diploid DNA patients had a significantly lower rate of disease recurrence (P=.0086) (hazard ratio [HR] 0.256). On multivariable analysis, patients with aneuploid tumors had a higher prostate-specific antigen failure rate (HR 5.13). Additionally, those with “excellent” dosimetry (V100 >90%; D90 >144 Gy) had a significantly lower recurrence rate (HR 0.25). All patients with aneuploid tumors and dosimetry classified as “nonexcellent” (V100 <90%; D90 <144 Gy) (5 of 5) had disease recurrence, compared with 40% of patients with aneuploid tumors and “excellent” dosimetry (8 of 15). In contrast, dosimetry did not affect the outcome for diploid patients. Conclusions: Using core biopsy material from archived paraffin blocks, DNA ploidy correctly classified the majority of failures and nonfailures in this study. The results suggest that DNA ploidy can be used as a

  17. Design and evaluation of protein expression in a recombinant plasmid encoding epitope gp 350/220 of the Epstein-Barr virus (EBV)

    Science.gov (United States)

    Himmah, Karimatul; Dluha, Nurul; Anyndita, Nadya V. M.; Rifa'i, Muhaimin; Widodo

    2017-05-01

    The Epstein - Barr virus (EBV) causes severe infections that may lead to cancers such as nasopharyngeal carcinoma. Development of effective EBV vaccines is necessary to prevent the virus spreading throughout the community. TheEBV has a surface protein gp 350/220, which serves as an antigen to help interact with host cells. Epitopes of the protein can potentially serve as bases for a vaccine. In a previous study, we have found a conserved epitope of gp 350/220 from all strains EBV through an in silico approach. The aim of this study is to design and overproduce a recombinant peptide of epitope gp 350/220 in E. coli. DNA encoding the conserved epitope was synthesized and cloned into plasmid pET-22b(+); the recombinant plasmid was transformed into E. coli strains DH5α and BL21. The transformed plasmid DNA was isolated and confirmed by restriction using XbaI and PstI enzymes followed by DNA sequencing. Protein expression was induced by isopropyl-D-thiogalactopyranoside (IPTG) with final concentrations of 0.1, 0.2, 1, and 2 mM in consecutive times. An osmotic shock method was used to isolate protein from periplasmic fraction of E. coli DH5α and BL21. The SDS-PAGE analysis was carried out to detect peptide target (3.4 kDa). Based on this result, the induction process did not work properly, and thus needs further investigation.

  18. Prevalence of Polyoma BK Virus (BKPyV), Epstein-Barr Virus (EBV) and Human Papilloma Virus (HPV) in Oropharyngeal Cancer.

    Science.gov (United States)

    Polz-Gruszka, Dorota; Morshed, Kamal; Jarzyński, Adrian; Polz-Dacewicz, Małgorzata

    2015-01-01

    The aim of this study was to analyze the prevalence of BK virus, Human Papillomavirus and Epstein-Barr virus in oropharyngeal cancer, and to test our hypothesis that BKV/HPV/EBV co-infection plays a role in oropharyngeal squamous cell carcinoma. The correlation between viral infection, OSCC, anatomic location, pre-treatment staging, evidence of metastases to lymph nodes, and grading was also investigated. The examination samples were collected from 62 patients from paraffin tissue blocks. Males (90.3%) with, smoking (83.9%) and alcohol abuse (67.7%) problems prevailed in the studied group. G2 histological type was recognized in 80.6% cases. T4 (77.4%) and N2 (56.5%) traits occurred in the majority of patients. No cases of metastasis were observed (M0 100%). HPV - 24.2%, EBV - 27.4% and BKV 17.7% were detected in the studied samples. We observed co-infection EBV/BKV in 8% of cases, HPV/BKV in 4.8%, and HPV/EBV in 9% cases. Only in two cases co-infection of all three viruses was found.

  19. Pretreatment antigen-specific immunity and regulation - association with subsequent immune response to anti-tumor DNA vaccination.

    Science.gov (United States)

    Johnson, Laura E; Olson, Brian M; McNeel, Douglas G

    2017-07-18

    Immunotherapies have demonstrated clinical benefit for many types of cancers, however many patients do not respond, and treatment-related adverse effects can be severe. Hence many efforts are underway to identify treatment predictive biomarkers. We have reported the results of two phase I trials using a DNA vaccine encoding prostatic acid phosphatase (PAP) in patients with biochemically recurrent prostate cancer. In both trials, persistent PAP-specific Th1 immunity developed in some patients, and this was associated with favorable changes in serum PSA kinetics. In the current study, we sought to determine if measures of antigen-specific or antigen non-specific immunity were present prior to treatment, and associated with subsequent immune response, to identify possible predictive immune biomarkers. Patients who developed persistent PAP-specific, IFNγ-secreting immune responses were defined as immune "responders." The frequency of peripheral T cell and B cell lymphocytes, natural killer cells, monocytes, dendritic cells, myeloid derived suppressor cells, and regulatory T cells were assessed by flow cytometry and clinical laboratory values. PAP-specific immune responses were evaluated by cytokine secretion in vitro, and by antigen-specific suppression of delayed-type hypersensitivity to a recall antigen in an in vivo SCID mouse model. The frequency of peripheral blood cell types did not differ between the immune responder and non-responder groups. Non-responder patients tended to have higher PAP-specific IL-10 production pre-vaccination (p = 0.09). Responder patients had greater preexisting PAP-specific bystander regulatory responses that suppressed DTH to a recall antigen (p = 0.016). While our study population was small (n = 38), these results suggest that different measures of antigen-specific tolerance or regulation might help predict immunological outcome from DNA vaccination. These will be prospectively evaluated in an ongoing randomized, phase II trial.

  20. Building essential biodiversity variables (EBVs) of species distribution and abundance at a global scale.

    Science.gov (United States)

    Kissling, W Daniel; Ahumada, Jorge A; Bowser, Anne; Fernandez, Miguel; Fernández, Néstor; García, Enrique Alonso; Guralnick, Robert P; Isaac, Nick J B; Kelling, Steve; Los, Wouter; McRae, Louise; Mihoub, Jean-Baptiste; Obst, Matthias; Santamaria, Monica; Skidmore, Andrew K; Williams, Kristen J; Agosti, Donat; Amariles, Daniel; Arvanitidis, Christos; Bastin, Lucy; De Leo, Francesca; Egloff, Willi; Elith, Jane; Hobern, Donald; Martin, David; Pereira, Henrique M; Pesole, Graziano; Peterseil, Johannes; Saarenmaa, Hannu; Schigel, Dmitry; Schmeller, Dirk S; Segata, Nicola; Turak, Eren; Uhlir, Paul F; Wee, Brian; Hardisty, Alex R

    2018-02-01

    aggregation of various raw data sources, data quality control, taxonomic name matching and statistical modelling of integrated data. We illustrate these steps with concrete examples from existing citizen science and professional monitoring projects, including eBird, the Tropical Ecology Assessment and Monitoring network, the Living Planet Index and the Baltic Sea zooplankton monitoring. The identified workflow steps are applicable to both terrestrial and aquatic systems and a broad range of spatial, temporal and taxonomic scales. They depend on clear, findable and accessible metadata, and we provide an overview of current data and metadata standards. Several challenges remain to be solved for building global EBV data products: (i) developing tools and models for combining heterogeneous, multi-source data sets and filling data gaps in geographic, temporal and taxonomic coverage, (ii) integrating emerging methods and technologies for data collection such as citizen science, sensor networks, DNA-based techniques and satellite remote sensing, (iii) solving major technical issues related to data product structure, data storage, execution of workflows and the production process/cycle as well as approaching technical interoperability among research infrastructures, (iv) allowing semantic interoperability by developing and adopting standards and tools for capturing consistent data and metadata, and (v) ensuring legal interoperability by endorsing open data or data that are free from restrictions on use, modification and sharing. Addressing these challenges is critical for biodiversity research and for assessing progress towards conservation policy targets and sustainable development goals. © 2017 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.

  1. Deregressed EBV as the response variable yield more reliable genomic predictions than traditional EBV in pure-bred pigs

    DEFF Research Database (Denmark)

    Ostersen, Tage; Christensen, Ole Fredslund; Henryon, Mark

    2011-01-01

    Background Genomic selection can be implemented by a multi-step procedure, which requires a response variable and a statistical method. For pure-bred pigs, it was hypothesised that deregressed estimated breeding values (EBV) with the parent average removed as the response variable generate higher...

  2. Shedding of HSV-1, HSV-2, CMV, and EBV in the saliva of hematopoietic stem cell transplant recipients at Fundación HOMI - Hospital de la Misericordia, Bogotá, D.C.

    Science.gov (United States)

    Bohórquez, Sonia P; Díaz, Juliana; Rincón, Claudia M; Estupiñán, Marcela; Chaparro, Mauricio; Low-Calle, Ana María; Castellanos, Jaime E

    2016-05-19

    Hematopoietic stem cell transplantation in pediatric patients is an alternative treatment for different diseases. The conditioning regimen for transplant predisposes recipients to the development of infections. Viral infections by herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), human cytomegalovirus (CMV), and Epstein-Barr virus (EBV), are the most common, and the leading cause of morbidity and mortality among these patients. These viruses lie dormant in various cell types and the reactivation of latent infections may lead to asymptomatic viral shedding in saliva. The detection of these viruses in secretions may contribute to understand the behavioral dynamics of these viral infections in transplanted patients, and to the early diagnosis of reactivation.  To assess HSV-1, HSV-2, CMV and EBV viral shedding in the saliva of patients admitted for hematopoietic stem cell transplantation at Fundación HOMI - Hospital de la Misericordia between January and November of 2012.  We evaluated stimulated saliva samples of 17 hematopoietic stem cell transplantation recipients weekly. We performed DNA extraction from saliva, and we evaluated the presence of DNA for HSV-1, HSV-2, CMV, and EBV by PCR.  While we detected HSV-2 and CMV DNA in the saliva of four patients, EBV DNA was detected in nine patients with leukopenia. In contrast, we did not detect HSV-1 DNA in saliva. Additionally, four out of the 17 patients showed a simultaneous shedding of CMV and EBV.  By conventional PCR, we demonstrated asymptomatic HSV-2, CMV, and EBV viral shedding in saliva, associated with leukopenia.

  3. Coinfection with Epstein–Barr Virus (EBV, Human Papilloma Virus (HPV and Polyoma BK Virus (BKPyV in Laryngeal, Oropharyngeal and Oral Cavity Cancer

    Directory of Open Access Journals (Sweden)

    Bartłomiej Drop

    2017-12-01

    Full Text Available Most research providing evidence for the role of oncogenic viruses in head and neck squamous cell carcinoma (SCC development is focused on one type of virus without analyzing possible interactions between two or more types of viruses. The aim of this study was to analyse the prevalence of co-infection with human papillomavirus (HPV, Epstein–Barr virus (EBV and polyoma BK virus (BKPyV in oral, oropharyngeal and laryngeal squamous cell carcinomas in Polish patients. The correlations between viral infection, SCC, demographic parameters, evidence of metastases and grading were also investigated. Fresh-frozen tumour tissue samples were collected from 146 patients with laryngeal, oropharyngeal and oral cancer. After DNA extraction, the DNA of the studied viruses was detected using polymerase chain rection (PCR assay. Males (87.7% with a history of smoking (70.6% and alcohol abuse (59.6% prevailed in the studied group. Histological type G2 was recognized in 64.4% cases. The patients were most frequently diagnosed with T2 stage (36.3% and with N1 stage (45.8%. Infection with at least two viruses was detected in 56.2% of patients. In this group, co-infection with HPV/EBV was identified in 34.1% of cases, EBV/BKV in 23.2%, HPV/BKV in 22.0%, and HPV/EBV/BKV in 20.7%. No difference of multiple infection in different locations of cancer was observed. The prevalence of poorly differentiated tumours (G3 was more frequent in co-infection with all three viruses than EBV or BKV alone. A significant correlation was observed between tumour dimensions (T and lymph-node involvement (N in co-infected patients compared to single infection. Further studies are necessary to clarify whether co-infection plays an important role in the initiation and/or progression of oncogenic transformation of oral, oropharyngeal and laryngeal epithelial cells.

  4. Coinfection with Epstein-Barr Virus (EBV), Human Papilloma Virus (HPV) and Polyoma BK Virus (BKPyV) in Laryngeal, Oropharyngeal and Oral Cavity Cancer.

    Science.gov (United States)

    Drop, Bartłomiej; Strycharz-Dudziak, Małgorzata; Kliszczewska, Ewa; Polz-Dacewicz, Małgorzata

    2017-12-19

    Most research providing evidence for the role of oncogenic viruses in head and neck squamous cell carcinoma (SCC) development is focused on one type of virus without analyzing possible interactions between two or more types of viruses. The aim of this study was to analyse the prevalence of co-infection with human papillomavirus (HPV), Epstein-Barr virus (EBV) and polyoma BK virus (BKPyV) in oral, oropharyngeal and laryngeal squamous cell carcinomas in Polish patients. The correlations between viral infection, SCC, demographic parameters, evidence of metastases and grading were also investigated. Fresh-frozen tumour tissue samples were collected from 146 patients with laryngeal, oropharyngeal and oral cancer. After DNA extraction, the DNA of the studied viruses was detected using polymerase chain rection (PCR) assay. Males (87.7%) with a history of smoking (70.6%) and alcohol abuse (59.6%) prevailed in the studied group. Histological type G2 was recognized in 64.4% cases. The patients were most frequently diagnosed with T2 stage (36.3%) and with N1 stage (45.8%). Infection with at least two viruses was detected in 56.2% of patients. In this group, co-infection with HPV/EBV was identified in 34.1% of cases, EBV/BKV in 23.2%, HPV/BKV in 22.0%, and HPV/EBV/BKV in 20.7%. No difference of multiple infection in different locations of cancer was observed. The prevalence of poorly differentiated tumours (G3) was more frequent in co-infection with all three viruses than EBV or BKV alone. A significant correlation was observed between tumour dimensions (T) and lymph-node involvement (N) in co-infected patients compared to single infection. Further studies are necessary to clarify whether co-infection plays an important role in the initiation and/or progression of oncogenic transformation of oral, oropharyngeal and laryngeal epithelial cells.

  5. EBV induces persistent NF-κB activation and contributes to survival of EBV-positive neoplastic T- or NK-cells.

    Directory of Open Access Journals (Sweden)

    Honami Takada

    Full Text Available Epstein-Barr virus (EBV has been detected in several T- and NK-cell neoplasms such as extranodal NK/T-cell lymphoma nasal type, aggressive NK-cell leukemia, EBV-positive peripheral T-cell lymphoma, systemic EBV-positive T-cell lymphoma of childhood, and chronic active EBV infection (CAEBV. However, how this virus contributes to lymphomagenesis in T or NK cells remains largely unknown. Here, we examined NF-κB activation in EBV-positive T or NK cell lines, SNT8, SNT15, SNT16, SNK6, and primary EBV-positive and clonally proliferating T/NK cells obtained from the peripheral blood of patients with CAEBV. Western blotting, electrophoretic mobility shift assays, and immunofluorescent staining revealed persistent NF-κB activation in EBV-infected cell lines and primary cells from patients. Furthermore, we investigated the role of EBV in infected T cells. We performed an in vitro infection assay using MOLT4 cells infected with EBV. The infection directly induced NF-κB activation, promoted survival, and inhibited etoposide-induced apoptosis in MOLT4 cells. The luciferase assay suggested that LMP1 mediated NF-κB activation in MOLT4 cells. IMD-0354, a specific inhibitor of NF-κB that suppresses NF-κB activation in cell lines, inhibited cell survival and induced apoptosis. These results indicate that EBV induces NF-κB-mediated survival signals in T and NK cells, and therefore, may contribute to the lymphomagenesis of these cells.

  6. Immunoglobulin gene analysis reveals 2 distinct cells of origin for EBV-positive and EBV-negative Burkitt lymphomas.

    Science.gov (United States)

    Bellan, Cristiana; Lazzi, Stefano; Hummel, Michael; Palummo, Nazzareno; de Santi, Margherita; Amato, Teresa; Nyagol, Joshua; Sabattini, Elena; Lazure, Thierry; Pileri, Stefano A; Raphael, Martine; Stein, Harald; Tosi, Piero; Leoncini, Lorenzo

    2005-08-01

    The normal counterpart of the neoplastic B cells in Burkitt lymphoma (BL) is still unclear. Based on immunoglobulin gene rearrangement studies, some authors suggest an origin from germinal center cells and others from memory B cells. However, most of these studies rely on cell lines or on a small series of cases. To help clarify the cell of origin of BL, semi-nested polymerase chain reaction (PCR) was performed to amplify the VDJ rearrangements of the immunoglobulin heavy chain (V(H)) genes, and the resultant amplificates were sequenced for comparison with known germline V(H) segments. The results of this approach revealed that all cases (15 endemic BL [eBL], 10 sporadic BL [sBL], and 6 AIDS-related BL) harbor mutated V(H) genes, with different mutation ranges among the 3 types of BL. The eBL and AIDS-related forms showed considerably higher mutation rates than the sBL form (5.1%, 5.4%, and 1.5%, respectively). The mutations in eBL and AIDS-related BL also showed signs of antigen selection, whereas no signs of antigen selection were found in sBL. Finally, after subcloning the amplificates, sequence analysis revealed no signs of ongoing mutations in any of the cases analyzed. Given that one of the main differences between eBL and AIDS-related BL on the one hand and sBL on the other hand is the association with Epstein-Barr virus (EBV), we compared EBV-positive and EBV-negative BLs independently of their geographic origin and HIV status. The differences in the number of somatic mutations and antigen selection were even more evident when this approach was used. According to our molecular results, it appears that EBV-positive and EBV-negative BL may originate from 2 distinct subsets of B cells, pointing to a particular role for the germinal-center reaction in the pathogenesis of these tumors. The different types of C-MYC translocation reported in BL may also be related to the different stages of B-cell maturation.

  7. Atypical prediagnosis Epstein-Barr virus serology restricted to EBV-positive Hodgkin lymphoma

    Science.gov (United States)

    Chang, Ellen T.; Ambinder, Richard F.; Lennette, Evelyne T.; Rubertone, Mark V.; Mann, Risa B.; Borowitz, Michael; Weir, Edward G.; Abbondanzo, Susan L.; Mueller, Nancy E.

    2012-01-01

    An altered anti–Epstein-Barr virus (EBV) serologic profile preceding diagnosis is associated with an increased risk of Hodgkin lymphoma. It is unknown whether this atypical pattern predicts Hodgkin lymphoma risk further subdivided by determination of EBV in tumor cells. A nested case-control study of 128 incident Hodgkin lymphoma cases and 368 matched controls from active-duty military personnel with archived serum in the US Department of Defense Serum Repository was conducted to determine whether a panel of anti-EBV antibody titers differed in EBV+ and EBV− Hodgkin lymphoma. Among 40 EBV+ Hodgkin lymphoma cases and matched controls, statistically significant increased risks were associated with elevated anti-EBV VCA IgG antibody titers (relative risk = 3.1; 95% confidence interval [CI], 1.1-8.7), and an anti–EBNA-1/anti–EBNA-2 antibody ratio ≤ 1.0 versus > 1.0 (relative risk = 4.7; 95% CI, 1.6-13.8). In contrast, no significant associations were found among 88 EBV− Hodgkin lymphoma cases relative to their matched controls. In case-case analysis, EBV+ disease was significantly associated with a low anti–EBNA-1/anti–EBNA-2 antibody ratio. This distinc-tive serologic response to EBV latent antigens, indicative of immune dysfunction in other clinical settings, is associated with an increased risk of developing EBV+ but not EBV− Hodgkin lymphoma. PMID:22972983

  8. Interaction of Epstein-Barr virus (EBV) with human B-lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Klein, George, E-mail: Georg.Klein@ki.se [Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology (MTC), Box 280, S171 77 Stockholm (Sweden); Klein, Eva; Kashuba, Elena [Karolinska Institutet, Department of Microbiology, Tumor and Cell Biology (MTC), Box 280, S171 77 Stockholm (Sweden)

    2010-05-21

    Epstein-Barr virus, EBV, and humans have a common history that reaches back to our primate ancestors. The virus co-evolved with man and has established a largely harmless and highly complex co-existence. It is carried as silent infection by almost all human adults. A serendipitous discovery established that it is the causative agent of infectious mononucleosis. Still, EBV became known first in 1964, in a rare, geographically prevalent malignant lymphoma of B-cell origin, Burkitt lymphoma BL. Its association with a malignancy prompted intensive studies and its capacity to immortalize B-lymphocytes in vitro was soon demonstrated. Consequently EBV was classified therefore as a potentially tumorigenic virus. Despite of this property however, the virus carrier state itself does not lead to malignancies because the transformed cells are recognized by the immune response. Consequently the EBV induced proliferation of EBV carrying B-lymphocytes is manifested only under immunosuppressive conditions. The expression of EBV encoded genes is regulated by the cell phenotype. The virus genome can be found in malignancies originating from cell types other than the B-lymphocyte. Even in the EBV infected B-cell, the direct transforming capacity is restricted to a defined window of differentiation. A complex interaction between virally encoded proteins and B-cell specific cellular proteins constitute the proliferation inducing program. In this short review we touch upon aspects which are the subject of our present work. We describe the mechanisms of some of the functional interactions between EBV encoded and cellular proteins that determine the phenotype of latently infected B-cells. The growth promoting EBV encoded genes are not expressed in the virus carrying BL cells. Still, EBV seems to contribute to the etiology of this tumor by modifying events that influence cell survival and proliferation. We describe a possible growth promoting mechanism in the genesis of Burkitt lymphoma

  9. EBV-positive diffuse large B-cell lymphoma of the elderly

    NARCIS (Netherlands)

    C.Y. Ok (Chi Young); T.G. Papathomas (Thomas); L.J. Medeiros (L. Jeffrey); K.H. Young (Ken)

    2013-01-01

    textabstractEpstein-Barr virus (EBV) positive diffuse large B-cell lymphoma (DLBCL) of the elderly, initially described in 2003, is a provisional entity in the 2008World Health Organization classification system and is defined as an EBV-positive monoclonal large B-cell proliferation that occurs in

  10. Ascorbic Acid Kills Epstein-Barr Virus (EBV) Positive Burkitt Lymphoma Cells and EBV Transformed B-Cells in Vitro, but not in Vivo

    Science.gov (United States)

    Shatzer, Amber N.; Espey, Michael G.; Chavez, Mayra; Tu, Hongbin; Levine, Mark; Cohen, Jeffrey I.

    2014-01-01

    Ascorbic acid has been shown to kill various cancer cell lines at pharmacologic concentrations. We found that Epstein-Barr virus (EBV)-positive Burkitt lymphoma (BL) cells were more susceptible to ascorbic acid-induced cell killing than EBV-negative BL cells or EBV-transformed lymphoblastoid cells (LCLs). Ascorbic acid did not induce apoptosis in any of the tested cells but did induce the production of reactive oxygen species and cell death. Previously, we showed that bortezomib, a proteasome inhibitor, induces cell death in LCLs and EBV-positive BL cells. We found that ascorbic acid is strongly antagonistic for ascorbic acid-induced cell death in LCLs and EBV-positive BL cells. Finally, ascorbic acid did not prolong survival of severe combined immunodefiency mice inoculated with LCLs either intraperitoneally or subcutaneously. Thus, while ascorbic acid was highly effective at killing EBV-positive BL cells and LCLs in vitro, it antagonized cell killing by bortezomib and was ineffective in an animal model. PMID:23067008

  11. Altered EBV viral load setpoint after HIV seroconversion is in accordance with lack of predictive value of EBV load for the occurrence of AIDS-related non-Hodgkin lymphoma

    NARCIS (Netherlands)

    Piriou, Erwan R.; van Dort, Karel; Nanlohy, Nening M.; Miedema, Frank; van Oers, Marinus H.; van Baarle, Debbie

    2004-01-01

    In contrast to the situation in the post-transplant setting, in HIV-infected individuals an elevated EBV load is not predictive of EBV-related malignancies. To study whether a high EBV load is already a normal situation early in HIV infection and is not related to a decrease in immune function over

  12. Epstein-Barr virus (EBV) reactivation is a frequent event after allogeneic stem cell transplantation (SCT) and quantitatively predicts EBV-lymphoproliferative disease following T-cell--depleted SCT

    NARCIS (Netherlands)

    van Esser, J W; van der Holt, B; Meijer, E; Niesters, H G; Trenschel, R; Thijsen, S F; van Loon, A M; Frassoni, F; Bacigalupo, A; Schaefer, U W; Osterhaus, A D; Gratama, J W; Löwenberg, B; Verdonck, L F; Cornelissen, J J

    2001-01-01

    Reactivation of the Epstein-Barr virus (EBV) after allogeneic stem cell transplantation (allo-SCT) may evoke a protective cellular immune response or may be complicated by the development of EBV-lymphoproliferative disease (EBV-LPD). So far, very little is known about the incidence, recurrence, and

  13. Emergence of CD4+ and CD8+ Polyfunctional T Cell Responses Against Immunodominant Lytic and Latent EBV Antigens in Children With Primary EBV Infection

    Directory of Open Access Journals (Sweden)

    Janice K. P. Lam

    2018-03-01

    Full Text Available Long term carriers were shown to generate robust polyfunctional T cell (PFC responses against lytic and latent antigens of Epstein-Barr virus (EBV. However, the time of emergence of PFC responses against EBV antigens, pattern of immunodominance and difference between CD4+ and CD8+ T cell responses during various stages of EBV infection are not clearly understood. A longitudinal study was performed to assess the development of antigen-specific PFC responses in children diagnosed to have primary symptomatic (infectious mononucleosis [IM] and asymptomatic (AS EBV infection. Evaluation of IFN-γ secreting CD8+ T cell responses upon stimulation by HLA class I-specific peptides of EBV lytic and latent proteins by ELISPOT assay followed by assessment of CD4+ and CD8+ PFC responses upon stimulation by a panel of overlapping EBV peptides for co-expression of IFN-γ, TNF-α, IL-2, perforin and CD107a by flow cytometry were performed. Cytotoxicity of T cells against autologous lymphoblastoid cell lines (LCLs as well as EBV loads in PBMC and plasma were also determined. Both IM and AS patients had elevated PBMC and plasma viral loads which declined steadily during a 12-month period from the time of diagnosis whilst decrease in the magnitude of CD8+ T cell responses toward EBV lytic peptides in contrast to increase toward latent peptides was shown with no significant difference between those of IM and AS patients. Both lytic and latent antigen-specific CD4+ and CD8+ T cells demonstrated polyfunctionality (defined as greater or equal to three functions concurrent with enhanced cytotoxicity against autologous LCLs and steady decrease in plasma and PBMC viral loads over time. Immunodominant peptides derived from BZLF1, BRLF1, BMLF1 and EBNA3A-C proteins induced the highest proportion of CD8+ as well as CD4+ PFC responses. Diverse functional subtypes of both CD4+ and CD8+ PFCs were shown to emerge at 6–12 months. In conclusion, EBV antigen-specific CD4+ and CD

  14. Bilateral adrenal EBV-associated smooth muscle tumors in a child with a natural killer cell deficiency

    OpenAIRE

    Shaw, Rachel K.; Issekutz, Andrew C.; Fraser, Robert; Schmit, Pierre; Morash, Barb; Monaco-Shawver, Linda; Orange, Jordan S.; Fernandez, Conrad V.

    2012-01-01

    EBV-associated smooth muscle tumors are found in immunocompromised patients, most commonly HIV/AIDS. We present a 12-year-old girl with the first documented case of EBV-related smooth muscle tumors in the presence of a rare classic NK cell deficiency. This sheds light on the role of NK cells in controlling EBV-related smooth muscle tumors.

  15. Epstein-Barr Viruses (EBVs) Deficient in EBV-Encoded RNAs Have Higher Levels of Latent Membrane Protein 2 RNA Expression in Lymphoblastoid Cell Lines and Efficiently Establish Persistent Infections in Humanized Mice.

    Science.gov (United States)

    Gregorovic, Goran; Boulden, Elizabeth A; Bosshard, Rachel; Elgueta Karstegl, Claudio; Skalsky, Rebecca; Cullen, Bryan R; Gujer, Cornelia; Rämer, Patrick; Münz, Christian; Farrell, Paul J

    2015-11-01

    Functions of Epstein-Barr virus (EBV)-encoded RNAs (EBERs) were tested in lymphoblastoid cell lines containing EBER mutants of EBV. Binding of EBER1 to ribosomal protein L22 (RPL22) was confirmed. Deletion of EBER1 or EBER2 correlated with increased levels of cytoplasmic EBV LMP2 RNA and with small effects on specific cellular microRNA (miRNA) levels, but protein levels of LMP1 and LMP2A were not affected. Wild-type EBV and EBER deletion EBV had approximately equal abilities to infect immunodeficient mice reconstituted with a human hematopoietic system. Copyright © 2015, Gregorovic et al.

  16. Serological profiling of the EBV immune response in Chronic Fatigue Syndrome using a peptide microarray.

    Directory of Open Access Journals (Sweden)

    Madlen Loebel

    Full Text Available Epstein-Barr-Virus (EBV plays an important role as trigger or cofactor for various autoimmune diseases. In a subset of patients with Chronic Fatigue Syndrome (CFS disease starts with infectious mononucleosis as late primary EBV-infection, whereby altered levels of EBV-specific antibodies can be observed in another subset of patients.We performed a comprehensive mapping of the IgG response against EBV comparing 50 healthy controls with 92 CFS patients using a microarray platform. Patients with multiple sclerosis (MS, systemic lupus erythematosus (SLE and cancer-related fatigue served as controls. 3054 overlapping peptides were synthesised as 15-mers from 14 different EBV proteins. Array data was validated by ELISA for selected peptides. Prevalence of EBV serotypes was determined by qPCR from throat washing samples.EBV type 1 infections were found in patients and controls. EBV seroarray profiles between healthy controls and CFS were less divergent than that observed for MS or SLE. We found significantly enhanced IgG responses to several EBNA-6 peptides containing a repeat sequence in CFS patients compared to controls. EBNA-6 peptide IgG responses correlated well with EBNA-6 protein responses. The EBNA-6 repeat region showed sequence homologies to various human proteins.Patients with CFS had a quite similar EBV IgG antibody response pattern as healthy controls. Enhanced IgG reactivity against an EBNA-6 repeat sequence and against EBNA-6 protein is found in CFS patients. Homologous sequences of various human proteins with this EBNA-6 repeat sequence might be potential targets for antigenic mimicry.

  17. Inhibitory activities of microalgal extracts against Epstein-Barr Virus (EBV antigen expression in lymphoblastoid cells

    Directory of Open Access Journals (Sweden)

    Koh Yih Yih

    2014-01-01

    Full Text Available The inhibitory activities of microalgal extracts against the expression of three EBV antigens, latent membrane protein (LMP1, Epstein-Barr nuclear antigen (EBNA1 and Z Epstein-Barr reactivation activator (ZEBRA were assessed by immunocytochemistry. The observation that the methanol extracts and their fractions from Ankistrodesmus convolutus, Synechococcus elongatus and Spirulina platensis exhibited inhibitory activity against EBV proteins in three Burkitt’s lymphoma cell lines at concentrations as low as 20 μg/ml suggests that microalgae could be a potential source of antiviral compounds against EBV.

  18. EBV-transformed lymphoblastoid cell lines as vaccines against cancer testis antigen-positive tumors.

    Science.gov (United States)

    Neumann, Frank; Kaddu-Mulindwa, Dominic; Widmann, Thomas; Preuss, Klaus-Dieter; Held, Gerhard; Zwick, Carsten; Roemer, Klaus; Pfreundschuh, Michael; Kubuschok, Boris

    2013-07-01

    EBV-transformed lymphoblastoid cell lines (LCL) are potent antigen-presenting cells. To investigate their potential use as cancer testis antigen (CTA) vaccines, we studied the expression of 12 cancer testis (CT) genes in 20 LCL by RT-PCR. The most frequently expressed CT genes were SSX4 (50 %), followed by GAGE (45 %), SSX1 (40 %), MAGE-A3 and SSX2 (25 %), SCP1, HOM-TES-85, MAGE-C1, and MAGE-C2 (15 %). NY-ESO-1 and MAGE-A4 were found in 1/20 LCL and BORIS was not detected at all. Fifteen of 20 LCL expressed at least one antigen, 9 LCL expressed ≥2 CT genes, and 7 of the 20 LCL expressed ≥4 CT genes. The expression of CT genes did not correlate with the length of in vitro culture, telomerase activity, aneuploidy, or proliferation state. While spontaneous expression of CT genes determined by real-time PCR and Western blot was rather weak in most LCL, treatment with DNA methyltransferase 1 inhibitor alone or in combination with histone deacetylase inhibitors increased CTA expression considerably thus enabling LCL to induce CTA-specific T cell responses. The stability of the CT gene expression over prolonged culture periods makes LCL attractive candidates for CT vaccines both in hematological neoplasias and solid tumors.

  19. HLA class II alleles and the presence of circulating Epstein-Barr virus DNA in greek patients with nasopharyngeal carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Karanikiotis, C. [424 Army General Hospital, Thessaloniki (Greece); Daniilidis, M.; Karyotis, N.; Nikolaou, A. [AHEPA Hospital, Aristotle Univ. of Thessaloniki School of Medicine (Greece); Bakogiannis, C. [Hygeia Hospital, Athens (Greece); Economopoulos, T. [' Attikon' Univ. Hospital, Athens (Greece); Murray, S. [Metropolitan Hospital, Athens (Greece); Papamichael, D. [Bank of Cyprus Oncology Center, Nicosia, Cyprus (Greece); Samantas, E. [' Agii Anargiri' Cancer Hospital, Athens (Greece); Skoura, L. [' Hippokration' Hospital, Thessaloniki (Greece); Tselis, N.; Zamboglou, N. [Dept. of Radiotherapy, Offenbach Hospital (Germany); Fountzilas, G. [' Papageorgiou' Hospital, Aristotle Univ. of Thessaloniki School of Medicine (Greece)

    2008-06-15

    Background and purpose: nasopharyngeal carcinoma (NPC) represents a seldom malignancy in most developed countries. Nevertheless, NPC receives an endemic form in concrete racial entities. The aims of this study were to detect the presence of Epstein-Barr virus DNA (EBV-DNA) in peripheral blood of NPC patients, to molecularly define human leukocyte antigens (HLA) DRB1*, DQA1* and DQB1* allele frequencies, and, finally, to determine whether the genetic predisposition of an individual to NPC depends on the liability to EBV infection. Patients and methods: a total of 101 patients of Hellenic origin and nationality, with histologically proven NPC, participated in this study. EBV-DNA detection was also applied in 66 patients with EBV-related malignancies (Hodgkin's [HL] and non-Hodgkin's lymphoma [NHL]) and infectious mononucleosis (IM), as well as in 80 healthy EBV-seropositive controls. Results: 81% of the NPC patients, 77.8% with HL, 72.2% with NHL, and 66.7% with IM were EBV-DNA positive, whereas the EBV genome was detected only in 15% of the healthy controls. These differences were statistically significant in all cases. Analysis of HLA class II antigens showed decreased frequency of the DRB1*07 (p = 0.003), DQA1*0103 (p = 0.002), and DQA1*0201 (p = 0.003) alleles among NPC patients. A significant association between the HLA-DR/DQ alleles and the presence of EBV-DNA in peripheral whole blood was not established. Conclusion: circulating EBV-DNA and specific HLA class II alleles may predispose to or protect from NPC. However, the results of this study suggest that the genetic predisposition of an individual to NPC is independent of the liability to EBV infection. (orig.)

  20. HLA class II alleles and the presence of circulating Epstein-Barr virus DNA in greek patients with nasopharyngeal carcinoma

    International Nuclear Information System (INIS)

    Karanikiotis, C.; Daniilidis, M.; Karyotis, N.; Nikolaou, A.; Bakogiannis, C.; Economopoulos, T.; Murray, S.; Papamichael, D.; Samantas, E.; Skoura, L.; Tselis, N.; Zamboglou, N.; Fountzilas, G.

    2008-01-01

    Background and purpose: nasopharyngeal carcinoma (NPC) represents a seldom malignancy in most developed countries. Nevertheless, NPC receives an endemic form in concrete racial entities. The aims of this study were to detect the presence of Epstein-Barr virus DNA (EBV-DNA) in peripheral blood of NPC patients, to molecularly define human leukocyte antigens (HLA) DRB1*, DQA1* and DQB1* allele frequencies, and, finally, to determine whether the genetic predisposition of an individual to NPC depends on the liability to EBV infection. Patients and methods: a total of 101 patients of Hellenic origin and nationality, with histologically proven NPC, participated in this study. EBV-DNA detection was also applied in 66 patients with EBV-related malignancies (Hodgkin's [HL] and non-Hodgkin's lymphoma [NHL]) and infectious mononucleosis (IM), as well as in 80 healthy EBV-seropositive controls. Results: 81% of the NPC patients, 77.8% with HL, 72.2% with NHL, and 66.7% with IM were EBV-DNA positive, whereas the EBV genome was detected only in 15% of the healthy controls. These differences were statistically significant in all cases. Analysis of HLA class II antigens showed decreased frequency of the DRB1*07 (p 0.003), DQA1*0103 (p = 0.002), and DQA1*0201 (p = 0.003) alleles among NPC patients. A significant association between the HLA-DR/DQ alleles and the presence of EBV-DNA in peripheral whole blood was not established. Conclusion: circulating EBV-DNA and specific HLA class II alleles may predispose to or protect from NPC. However, the results of this study suggest that the genetic predisposition of an individual to NPC is independent of the liability to EBV infection. (orig.)

  1. CD95 (Fas) ligand expression of Epstein-Barr virus (EBV)-infected lymphocytes: a possible mechanism of immune evasion in chronic active EBV infection.

    Science.gov (United States)

    Ohshima, K; Suzumiya, J; Sugihara, M; Nagafuchi, S; Ohga, S; Kikuchi, M

    1999-01-01

    The Epstein-Barr virus (EBV) induces infectious mononucleosis (IM) and can be associated with chronic active EBV infection (CAEBV). Cytotoxic T lymphocytes (CTL) play an important role in excluding EBV-infected cells. Two cytotoxic mechanisms of CTL have been demonstrated: one perforin/granzyme-based and the other Fas (CD95)/Fas ligand (FasL)-based. To clarify these two pathways in CAEBV, we analyzed six patients with CAEBV and four patients with IM using immunohistochemical staining of the lymph nodes. In both CAEBV and IM, CD8+ T-cells increased in number, but CD56+ natural killer cells were rare. In four of six cases with CAEBV, approximately half the lymphocytes were positive for T cell-restricted intracellular antigens (TIA-1), which were recognized by the cytolytic granules of CTL. In IM, the number of TIA-1 positive cells was smaller than that in CAEBV. Fas-positive lymphocytes were frequently encountered in both CAEBV and IM. However, FasL-positive lymphocytes increased in three of six patients with CAEBV, but not in patients with IM. Except for one case with CAEBV, the number of perforin- and/or granzyme-positive cells was small in number in both CAEBV and IM cases. In double-staining FasL and EBV in situ hybridization, FasL-positive EBV-infected lymphocytes were detected in CAEBV but not in IM. In CAEBV, the Fas/FasL pathway and not perforin pathways appears to play an important role in the pathogenesis. The data suggest that EBV-infected lymphocytes may evade immune attack through the expression of FasL.

  2. Building essential biodiversity variables (EBVs) of species distribution and abundance at a global scale

    NARCIS (Netherlands)

    Kissling, W.D.; Ahumada, J.A.; Bowser, A.; Fernandez, M.; Fernández, N.; Garcia, E.A.; Guralnick, R.P.; Isaac, N.J.B.; Kelling, S.; Los, W.; McRae, L.; Mihoub, J.-B.; Obst, M.; Santamaria, M.; Skidmore, A.K.; Williams, K.J.; Agosti, D.; Amariles, D.; Arvanitidis, C.; Bastin, L.; De Leo, F.; Egloff, W.; Elith, J.; Hobern, D.; Martin, D.; Pereira, H.M.; Pesole, G.; Peterseil, J.; Saarenmaa, H.; Schigel, D.; Schmeller, D.S.; Segata, N.; Turak, E.; Uhlir, P.F.; Wee, B.; Hardisty, A.R.

    2018-01-01

    Much biodiversity data is collected worldwide, but it remains challenging to assemble the scattered knowledge for assessing biodiversity status and trends. The concept of Essential Biodiversity Variables (EBVs) was introduced to structure biodiversity monitoring globally, and to harmonize and

  3. High incidence of Epstein-Barr virus (EBV)-positive Hodgkin lymphoma and Hodgkin lymphoma-like B-cell lymphoproliferations with EBV latency profile 2 in children with interleukin-2-inducible T-cell kinase deficiency.

    Science.gov (United States)

    Bienemann, Kirsten; Borkhardt, Arndt; Klapper, Wolfram; Oschlies, Ilske

    2015-11-01

    Interleukin-2-inducible T-cell kinase (ITK) deficiency is an inherited T-cell deficiency characterized by the development of Epstein-Barr virus (EBV)-associated lymphoproliferations. We aimed to describe the histopathological features of lymphoproliferative processes arising in ITK deficiency, and to compare them with lymphoproliferations in otherwise immunocompromised patients. We revised the histopathological diagnoses of 12 biopsies of lymphoproliferations from seven ITK-deficient children according to the World Health Organization criteria, and determined the EBV latency types and lytic activity by staining for EBV-encoded small RNA, latent membrane protein 1, EBV nuclear antigen 2, and ZEBRA. We found polymorphic and borderline polymorphic to monomorphic B-cell lymphoproliferations with variable contents in large cells (five cases), a Hodgkin-like B-cell proliferation (one case), and classic mixed-cellularity Hodgkin lymphoma (six cases). All cases (12/12) were EBV-positive. The Hodgkin lymphoma-like and Hodgkin lymphoma, and all but one polymorphic B-cell lymphoproliferation, showed EBV latency type 2, as observed in classic EBV-positive Hodgkin lymphoma. The 100% EBV association, the high percentage of EBV-positive classic Hodgkin lymphoma and Hodgkin-like B-cell proliferations and the predominance of EBV latency type 2 even in polymorphic lesions are the main features of lymphoproliferations in patients with ITK deficiency, and suggest a unique pathomechanism of lymphomagenesis in this T-cell immunodeficiency. © 2015 John Wiley & Sons Ltd.

  4. Changing patterns in the Epstein-Barr virus (EBV)and Hodgkin lymphoma association in Tunisia.

    Science.gov (United States)

    Dhiab, Myriam Ben; Ziadi, Sonia; Saad, Hanene; Louhichi, Teheni; Trimeche, Mounir

    2016-09-01

    We compared the features of the Epstein-Barr virus (EBV) and Hodgkin lymphoma (HL) association in Tunisia in two periods of time, 1991-2001 (111 cases) and 2002-2012 (122 cases). The investigation of the EBV status by EBER in situ hybridization showed a significant decrease in the prevalence of EBV-positive HL from 69.3 % for the period 1991-2001 to 40.1 % for the 2002-2012 period (p = 0.00001). EBV positivity has decreased in all age groups but was more pronounced among young patients, in the 15-24-year age group (46.1 vs 10.3 %, p = 0.003), in the 25-34-year age group (56.2 vs 25 %, p = 0.04), and among children (88.4 vs 59.2 %, p = 0.01). This decrease in EBV-positive HL over time contrasted with a remarkable increase in EBV-negative HL in young adults aged 15-34 years (51.2 vs 83 %; p = 0.001), especially among women (59.1 vs 91.2 %; p = 0.01). The decrease in EBV-positive HL over time concerns particularly the nodular sclerosis histological subtype (69.2 vs 31.6 %, p = 0.000001). These results indicate that the epidemiology of HL and its association with EBV are changing over time, with a trend toward a Western profile, and point toward the emergence of other environmental causative factors, especially among young women, which remain to be identified.

  5. Epstein-Barr virus DNA loads in adult human immunodeficiency virus type 1-infected patients receiving highly active antiretroviral therapy

    Science.gov (United States)

    Ling, Paul D.; Vilchez, Regis A.; Keitel, Wendy A.; Poston, David G.; Peng, Rong Sheng; White, Zoe S.; Visnegarwala, Fehmida; Lewis, Dorothy E.; Butel, Janet S.

    2003-01-01

    Patients with human immunodeficiency virus type 1 (HIV-1) infection are at high risk of developing Epstein-Barr virus (EBV)-associated lymphoma. However, little is known of the EBV DNA loads in patients receiving highly active antiretroviral therapy (HAART). Using a real-time quantitative polymerase chain reaction assay, we demonstrated that significantly more HIV-1-infected patients receiving HAART than HIV-1-uninfected volunteers had detectable EBV DNA in blood (57 [81%] of 70 vs. 11 [16%] of 68 patients; P=.001) and saliva (55 [79%] of 68 vs. 37 [54%] of 68 patients; P=.002). The mean EBV loads in blood and saliva samples were also higher in HIV-1-infected patients than in HIV-1-uninfected volunteers (P=.001). The frequency of EBV detection in blood was associated with lower CD4+ cell counts (P=.03) among HIV-1-infected individuals, although no differences were observed in the EBV DNA loads in blood or saliva samples in the HIV-1-infected group. Additional studies are needed to determine whether EBV-specific CD4+ and CD8+ cells play a role in the pathogenesis of EBV in HIV-1-infected patients receiving HAART.

  6. EBV-Associated Lymphoproliferative Disorder and Hemophagocytic Lymphohistiocytosis in a Patient with Severe Celiac Disease

    Directory of Open Access Journals (Sweden)

    John Jacob Kinross-Wright

    2018-01-01

    Full Text Available Background. Epstein-Barr virus- (EBV- associated lymphoproliferative disease (LPD is a rare condition, usually occurring in immunocompromised patients. We report a case of EBV-associated LPD in a patient with severe celiac disease, the first report to describe this syndrome in a patient with this diagnosis. Case Summary. A 69-year-old Caucasian woman with recent diagnosis of celiac sprue presented to our hospital with persistent diarrhea, abdominal pain, weight loss, and fatigue despite adherence to gluten-free diet for a number of weeks prior to presentation. She underwent evaluation for occult malignancy and was found to have diffuse intra-abdominal mesenteric lymphadenopathy on CT scan. Biopsy of mesenteric nodes revealed an EBV positive, CD20 positive mixed lymphoproliferative process with T-cell predominance, but without a monoclonal cell population felt to be consistent with EBV-associated LPD. Bone marrow biopsy revealed hemophagocytic lymphohistiocytosis, complicating her course. She was treated with steroids and rituximab but continued to decline, eventually developing MSSA bacteremia and succumbing to her disease. Conclusion. To our knowledge, this is the first report of the constellation of celiac sprue, EBV-associated LPD, and hemophagocytic lymphohistiocytosis. Providers caring for patients with severe, uncontrolled celiac disease and adenopathy should consider EBV-associated LPD.

  7. Coinfection with EBV/CMV and other respiratory agents in children with suspected infectious mononucleosis

    Directory of Open Access Journals (Sweden)

    Wei Cong

    2010-09-01

    Full Text Available Abstract Background Numerous studies have shown that Epstein-Barr virus (EBV and cytomegalovirus (CMV can infect immunocompetent patients simultaneously with other agents. Nonetheless, multiple infections with other agents in EBV/CMV-infected children have received little attention. We conducted a retrospective study of children with suspected infectious mononucleosis. Peripheral blood samples were analyzed by indirect immunofluorescence to detect EBV, CMV and other respiratory agents including respiratory syncytial virus; adenovirus; influenza virus types A and B; parainfluenza virus types 1, 2 and 3; Chlamydia pneumoniae and Mycoplasma pneumoniae. A medical history was collected for each child. Results The occurrence of multipathogen infections was 68.9%, 81.3% and 63.6% in the children with primary EBV, CMV or EBV/CMV, respectively, which was significantly higher than that in the past-infected group or the uninfected group (p C. pneumoniae in children with primary infection was as high as 50%, significantly higher than in the other groups (p Conclusion Our study suggests that there is a high incidence of multipathogen infections in children admitted with EBV/CMV primary infection and that the distribution of these pathogens is not random.

  8. Epstein-Barr virus (EBV infection in Chinese children: a retrospective study of age-specific prevalence.

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    Geng Xiong

    Full Text Available BACKGROUND: Epstein-Barr Virus (EBV is a globally prevalent herpesvirus associated with infectious mononucleosis and many malignancies. The survey on EBV prevalence appears to be important to study EBV-related diseases and determine when to administer prophylactic vaccine. The purpose of this retrospective study was to collect baseline information about the prevalence of EBV infection in Chinese children. METHODOLOGY/PRINCIPAL FINDING: We collected 1778 serum samples from healthy children aged 0 to 10, who were enrolled in conventional health and nutrition examinations without any EBV-related symptom in 2012 and 2013 in North China (n = 973 and South China (n = 805. We detected four EBV-specific antibodies, i.e., anti-VCA-IgG and IgM, anti-EBNA-IgG and anti-EA-IgG, by ELISA, representing all of the phases of EBV infection. The overall EBV seroprevalence in samples from North and South China were 80.78% and 79.38% respectively. The EBV seropositivity rates dropped slightly at age 2, and then increased gradually with age. The seroprevalence became stabilized at over 90% after age 8. In this study, the seroprevalence trends between North and South China showed no difference (P>0.05, and the trends of average antibody concentrations were similar as well (P>0.05. CONCLUSIONS/SIGNIFICANCE: EBV seroprevalence became more than 50% before age 3 in Chinese children, and exceed 90% after age 8. This study can be helpful to study the relationship between EBV and EBV-associated diseases, and supportive to EBV vaccine development and implementation.

  9. Frequent monitoring of Epstein-Barr virus DNA load in unfractionated whole blood is essential for early detection of posttransplant lymphoproliferative disease in high-risk patients

    NARCIS (Netherlands)

    Stevens, SJC; Verschuuren, EAM; Pronk, [No Value; van der Bij, W; Harmsen, MC; The, TH; Meijer, CJLM; van den Brule, AJC; Middeldorp, JM

    2001-01-01

    Posttransplant lymphoproliferative disease (PTLD) is a frequent and severe Epstein-Barr virus (EBV)-associated complication in transplantation recipients that is caused by iatrogenic suppression of T-cell function, The diagnostic value of weekly EBV DNA load monitoring was investigated in

  10. Viral induction and targeted inhibition of galectin-1 in EBV+ posttransplant lymphoproliferative disorders.

    Science.gov (United States)

    Ouyang, Jing; Juszczynski, Przemyslaw; Rodig, Scott J; Green, Michael R; O'Donnell, Evan; Currie, Treeve; Armant, Myriam; Takeyama, Kunihiko; Monti, Stefano; Rabinovich, Gabriel A; Ritz, Jerome; Kutok, Jeffery L; Shipp, Margaret A

    2011-04-21

    Posttransplant lymphoproliferative disorders (PTLDs) are potentially fatal, EBV-driven B-cell malignancies that develop in immunocompromised solid organ or hematopoietic stem cell recipients. In PTLD, the expression of EBV proteins, including latent membrane protein 1 (LMP1) and LMP2A, viral immune evasion strategies, and impaired host immune surveillance foster the proliferation of EBV-transformed B cells. Current PTLD treatment strategies include reduction of immunosuppression, which increases the risk of graft rejection, anti-CD20 treatment, combination chemotherapy, and administration of EBV-specific cytotoxic T cells. In the present study, we report that EBV-transformed lymphoblastoid B-cell lines (LCLs) and primary PTLDs overexpress galectin-1 (Gal1), a carbohydrate-binding lectin that induces tolerogenic dendritic cells and triggers the selective apoptosis of CD4(+) Th1 and Th17 cells and cytotoxic T cells. In transcriptional reporter assays, LMP2A and LMP1 each increased Gal1-driven luciferase expression, and the combination of LMP2A and LMP1 was additive. In addition, small interfering RNA (siRNA)-mediated depletion of LMP2A decreased Gal1 protein abundance in EBV-transformed LCLs. Gal1 expression in LCLs was dependent on both activating protein 1 (AP-1) and PI3K. A newly developed neutralizing Gal1 mAb selectively inhibited Gal1-mediated apoptosis of EBV-specific CD8(+) T cells. Given the tolerogenic and immunosuppressive function of Gal1, antibody-mediated Gal1 neutralization may represent a novel immunotherapeutic strategy for PTLD and other Gal1-expressing tumors.

  11. Identification of potential hot spots in the carboxy-terminal part of the Epstein-Barr virus (EBV) BNLF-1 gene in both malignant and benign EBV-associated diseases

    DEFF Research Database (Denmark)

    Sandvej, K; Peh, S C; Andresen, B S

    1994-01-01

    In this study, we have sequenced the C-terminal part of the Epstein-Barr virus (EBV)-BNLF-1 gene encoding for the latent membrane protein-1 from tissues of EBV-positive Danish Hodgkin's disease (HD) and of Danish and Malaysian peripheral T-cell lymphomas (PTLs) and from tonsils of Danish infectio...... of the CAO-BNLF-1 gene do not characterize a new EBV type A substrain. Rather, some of the positions of single base mutations and the 30-bp deletion are hot spots that may have mutated independently through the evolution of EBV strains....

  12. Ganetespib, an HSP90 inhibitor, kills Epstein-Barr virus (EBV)-infected B and T cells and reduces the percentage of EBV-infected cells in the blood.

    Science.gov (United States)

    Shatzer, Amber; Ali, Mir A; Chavez, Mayra; Dowdell, Kennichi; Lee, Min-Jung; Tomita, Yusuke; El-Hariry, Iman; Trepel, Jane B; Proia, David A; Cohen, Jeffrey I

    2017-04-01

    HSP90 inhibitors have been shown to kill Epstein-Barr virus (EBV)-infected cells by reducing the level of EBV EBNA-1 and/or LMP1. We treated virus-infected cells with ganetespib, an HSP90 inhibitor currently being evaluated in multiple clinical trials for cancer and found that the drug killed EBV-positive B and T cells and reduced the level of both EBV EBNA-1 and LMP1. Treatment of cells with ganetespib also reduced the level of pAkt. Ganetespib delayed the onset of EBV-positive lymphomas and prolonged survival in SCID mice inoculated with one EBV-transformed B-cell line, but not another B-cell line. The former cell line showed lower levels of EBNA-1 after treatment with ganetespib in vitro. Treatment of a patient with T-cell chronic active EBV with ganetespib reduced the percentage of EBV-positive cells in the peripheral blood. These data indicate that HSP90 inhibitors may have a role in the therapy of certain EBV-associated diseases.

  13. Coxsackie Myocarditis and Hepatitis with Reactivated Epstein-Bar Virus (EBV): A Case Report.

    Science.gov (United States)

    Atti, Varunsiri; Anderson, Nathan M; Day, Mathew B

    2017-02-15

    BACKGROUND Myocarditis, defined as inflammation of myocardial tissue of the heart, is an uncommon cardiac presentation and is due to a variety of causes. It affects 1% of the US population, 50% of which is caused by coxsackie B virus. Cardiac tissue is the prime target, and destruction of myocardium results in cardiac failure with fluid overload. CASE REPORT Our patient was a 57-year-old woman with fever, headache, neck pain, and generalized malaise. Her white blood cell count was 13×10³ cells/mm³. Interestingly, lumbar puncture ruled out meningitis. An echocardiogram to evaluate elevated troponin revealed an ejection fraction of 30% with severe left ventricular global hypokinesis without valvular vegetations consistent with new-onset systolic heart failure. Cardiac MRI showed a small pericardial effusion with bilateral pleural effusion. As she continued to be febrile, a viral panel was ordered, revealing coxsackie B4 antibody titer of 1: 640 (reference: >1: 32 indicates recent infection) with positive Epstein-Barr virus deoxyribonucleic acid by PCR, consistent with viral myocarditis. CONCLUSIONS Coxsackie B virus myocarditis is rarely recognized and reported by the general internist in clinical practice, so we would like present our experience with an interesting clinical presentation of the viral prodrome. An estimated 95% people in the US are infected with Epstein-Barr virus by adulthood, but it remains dormant in memory B lymphocytes. Recirculation of these B cells in lymphoid tissue stimulated by antigens, which in our case is coxsackie B virus; they differentiate into plasma cells, and the production of Z Epstein-Barr replication activator protein (ZEBRA) increases viral replication, thus explaining the positive EBV DNA measured by PCR.

  14. Chronic active EBV infection: the experience of the Samsung Medical Center in South Korea.

    Science.gov (United States)

    Lee, Tae-Hee; Ko, Young-Hyeh

    Chronic active EBV infection (CAEBV) of T-cell or NK-cell type is an EBV+ polyclonal, oligoclonal or often monoclonal lymphoproliferative disorder (LPD) recognized as representing the spectrum of EBV-associated T-cell and NK-cell LPD with different clinical presentations; one systemic and two cutaneous disorders including hydroa vacciniforme-like T-cell LPD and mosquito bite hypersensitivity. The systemic form of the disease is characterized by fever, persistent hepatitis, hepatosplenomegaly and lymphadenopathy, which shows varying degrees of clinical severity depending on the immune response of the host and the EBV viral load. We described the clinicopathological findings of two children with CAEBV with a brief review of the literature. Recognition of the disease is important for adequate management of the patient. EBV analysis should be included in the principal diagnostic tests for febrile children. Copyright © 2015 Hospital Infantil de México Federico Gómez. Publicado por Masson Doyma México S.A. All rights reserved.

  15. EBV tegument protein BNRF1 disrupts DAXX-ATRX to activate viral early gene transcription.

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    Kevin Tsai

    2011-11-01

    Full Text Available Productive infection by herpesviruses involve the disabling of host-cell intrinsic defenses by viral encoded tegument proteins. Epstein-Barr Virus (EBV typically establishes a non-productive, latent infection and it remains unclear how it confronts the host-cell intrinsic defenses that restrict viral gene expression. Here, we show that the EBV major tegument protein BNRF1 targets host-cell intrinsic defense proteins and promotes viral early gene activation. Specifically, we demonstrate that BNRF1 interacts with the host nuclear protein Daxx at PML nuclear bodies (PML-NBs and disrupts the formation of the Daxx-ATRX chromatin remodeling complex. We mapped the Daxx interaction domain on BNRF1, and show that this domain is important for supporting EBV primary infection. Through reverse transcription PCR and infection assays, we show that BNRF1 supports viral gene expression upon early infection, and that this function is dependent on the Daxx-interaction domain. Lastly, we show that knockdown of Daxx and ATRX induces reactivation of EBV from latently infected lymphoblastoid cell lines (LCLs, suggesting that Daxx and ATRX play a role in the regulation of viral chromatin. Taken together, our data demonstrate an important role of BNRF1 in supporting EBV early infection by interacting with Daxx and ATRX; and suggest that tegument disruption of PML-NB-associated antiviral resistances is a universal requirement for herpesvirus infection in the nucleus.

  16. EBV Tegument Protein BNRF1 Disrupts DAXX-ATRX to Activate Viral Early Gene Transcription

    Science.gov (United States)

    Tsai, Kevin; Thikmyanova, Nadezhda; Wojcechowskyj, Jason A.; Delecluse, Henri-Jacques; Lieberman, Paul M.

    2011-01-01

    Productive infection by herpesviruses involve the disabling of host-cell intrinsic defenses by viral encoded tegument proteins. Epstein-Barr Virus (EBV) typically establishes a non-productive, latent infection and it remains unclear how it confronts the host-cell intrinsic defenses that restrict viral gene expression. Here, we show that the EBV major tegument protein BNRF1 targets host-cell intrinsic defense proteins and promotes viral early gene activation. Specifically, we demonstrate that BNRF1 interacts with the host nuclear protein Daxx at PML nuclear bodies (PML-NBs) and disrupts the formation of the Daxx-ATRX chromatin remodeling complex. We mapped the Daxx interaction domain on BNRF1, and show that this domain is important for supporting EBV primary infection. Through reverse transcription PCR and infection assays, we show that BNRF1 supports viral gene expression upon early infection, and that this function is dependent on the Daxx-interaction domain. Lastly, we show that knockdown of Daxx and ATRX induces reactivation of EBV from latently infected lymphoblastoid cell lines (LCLs), suggesting that Daxx and ATRX play a role in the regulation of viral chromatin. Taken together, our data demonstrate an important role of BNRF1 in supporting EBV early infection by interacting with Daxx and ATRX; and suggest that tegument disruption of PML-NB-associated antiviral resistances is a universal requirement for herpesvirus infection in the nucleus. PMID:22102817

  17. Primary EBV infection induces an expression profile distinct from other viruses but similar to hemophagocytic syndromes.

    Directory of Open Access Journals (Sweden)

    Samantha K Dunmire

    Full Text Available Epstein-Barr Virus (EBV causes infectious mononucleosis and establishes lifelong infection associated with cancer and autoimmune disease. To better understand immunity to EBV, we performed a prospective study of natural infection in healthy humans. Transcriptome analysis defined a striking and reproducible expression profile during acute infection but no lasting gene changes were apparent during latent infection. Comparing the EBV response profile to multiple other acute viral infections, including influenza A (influenza, respiratory syncytial virus (RSV, human rhinovirus (HRV, attenuated yellow fever virus (YFV, and Dengue fever virus (DENV, revealed similarity only to DENV. The signature shared by EBV and DENV was also present in patients with hemophagocytic syndromes, suggesting these two viruses cause uncontrolled inflammatory responses. Interestingly, while EBV induced a strong type I interferon response, a subset of interferon induced genes, including MX1, HERC5, and OAS1, were not upregulated, suggesting a mechanism by which viral antagonism of immunity results in a profound inflammatory response. These data provide an important first description of the response to a natural herpesvirus infection in humans.

  18. Correlation between ebv co-infection and HPV16 genome integrity in Tunisian cervical cancer patients

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    Saloua Kahla

    2012-06-01

    Full Text Available Infection with high risk Human papillomavirus (HR-HPV is necessary but not sufficient to cause cervical carcinoma. This study explored whether multiple HR-HPV or coinfection with Epstein-Barr virus (EBV influence the integration status of HPV16 genome. The presence and typing of HPV in a series of 125 cervical specimens were assessed by polymerase chain reaction (PCR using the specific primers for the HPV L1 region. As for EBV infection, the viral EBNA1 gene was used for its detection through PCR amplification. Disruption of the HPV E2 gene was assessed by amplification of the entire E2 gene with single set of primers, while E2 transcripts were evaluated by a reverse transcription PCR method (RT-PCR. The overall prevalence of HPVDNA was of 81.8% in cervical cancers versus 26.9% in benign lesions. In HPV positive cases, HPV16 and HPV18 were the most prevalent types, followed by HPV types 33, 31. EBV EBNA1 prevalence was statistically more frequent in cervical carcinomas than in benign lesions (29.5%, vs 9.6%; P=0.01. No viral infection was detected in healthy control women. The uninterrupted E2 gene was correlated with the presence of E2 transcripts originating from the HPV episomal forms. It was observed that integration was more common in HPV18 and EBV coinfection. The presence of EBV caused a five-fold [OR= 5; CI= 1.15-21.8; P = 0.04] increase in the risk of HPV16 genome integration in the host genome. This study indicates that EBV infection is acting as a cofactor for induction of cervical cancer by favoring HPVDNA integration.

  19. Direct Epstein-Barr virus (EBV) typing on peripheral blood mononuclear cells: no association between EBV type 2 infection or superinfection and the development of acquired immunodeficiency syndrome-related non-Hodgkin's lymphoma

    NARCIS (Netherlands)

    van Baarle, D.; Hovenkamp, E.; Kersten, M. J.; Klein, M. R.; Miedema, F.; van Oers, M. H.

    1999-01-01

    In the literature, a correlation has been suggested between the occurrence of acquired immunodeficiency syndrome (AIDS)-related non-Hodgkin's lymphomas (NHL) and Epstein-Barr virus (EBV) type 2 infection. To further investigate a possible role for EBV type 2 infection in the development of AIDS-NHL,

  20. Dysfunctional Epstein-Barr virus (EBV)-specific CD8(+) T lymphocytes and increased EBV load in HIV-1 infected individuals progressing to AIDS-related non-Hodgkin lymphoma

    NARCIS (Netherlands)

    van Baarle, D; Hovenkamp, E; Callan, M F; Wolthers, K C; Kostense, S; Tan, L C; Niesters, H G; Osterhaus, A D; McMichael, A J; van Oers, M H; Miedema, F

    2001-01-01

    Acquired immunodeficiency syndrome-related non-Hodgkin lymphomas (AIDS-NHL) are thought to arise because of loss of Epstein-Barr Virus (EBV)-specific cellular immunity. Here, an investigation was done to determine whether cellular immunity to EBV is lost because of physical loss or dysfunction of

  1. Dysfunctional Epstein-Barr virus (EBV)-specific CD8(+) T lymphocytes and increased EBV load in HIV-1 infected individuals progressing to AIDS-related non-Hodgkin lymphoma

    NARCIS (Netherlands)

    D. van Baarle (Debbie); F. Miedema (Frank); E. Hovenkamp (Egbert); M.F.C. Callan (Margareth); K.C. Wolthers (Katja); S. Kostense; L.C. Tan; H.G.M. Niesters (Bert); A.D.M.E. Osterhaus (Albert); A.J. McMichael (Andrew); M.H.J. van Oers (Marinus)

    2001-01-01

    textabstractAcquired immunodeficiency syndrome-related non-Hodgkin lymphomas (AIDS-NHL) are thought to arise because of loss of Epstein-Barr Virus (EBV)-specific cellular immunity. Here, an investigation was done to determine whether cellular immunity to EBV is lost because of physical loss or

  2. Development of a robust, higher throughput green fluorescent protein (GFP)-based Epstein-Barr Virus (EBV) micro-neutralization assay.

    Science.gov (United States)

    Lin, Rui; Heeke, Darren; Liu, Hui; Rao, Eileen; Marshall, Jason D; Chio, Vera; Cataniag, Floro; Yu, Li; Zuo, Fengrong; McCarthy, Michael P

    2017-09-01

    The goal of most prophylactic vaccines is to elicit robust and effective neutralizing antibodies against the human pathogen target. The titer of neutralizing antibodies to Epstein-Barr Virus (EBV) is a useful biomarker for evaluating EBV vaccines. Here, the development and optimization of a 96-well micro-neutralization fluorescent imaging assay (FIA) using an EBV virus-encoding green fluorescent protein (GFP) to infect adherent EBV recipient cells is reported. The conditions were optimized for generating reproducible EBV-GFP virus, for maintaining viral infectivity for months, and for efficient viral infection of recipient cell culture. The utility of the EBV-GFP FIA neutralization assay was demonstrated in a mouse study of an investigational adjuvanted EBV gp350 subunit vaccine. This assay confirmed the generation of high titers of anti-EBV-neutralizing antibodies which correlated well with the established Raji cell-based flow cytometry-based EBV neutralization assay, as well as with anti-gp350 IgG titers. In naturally infected EBV+ human serum samples, a good correlation between anti-gp350 IgG ELISA titer and EBV-GFP FIA neutralization antibody titer was also observed. Taken together, these results demonstrate the establishment of a scalable high throughput EBV-GFP FIA micro-neutralization assay suitable to measure humoral EBV vaccine response in a large-scale human trial. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Frequency of Epstein-Barr virus DNA sequences in human gliomas

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    Renata Fragelli Fonseca

    Full Text Available CONTEXT AND OBJECTIVE: The Epstein-Barr virus (EBV is the most common cause of infectious mononucleosis and is also associated with several human tumors, including Burkitt's lymphoma, Hodgkin's lymphoma, some cases of gastric carcinoma and nasopharyngeal carcinoma, among other neoplasms. The aim of this study was to screen 75 primary gliomas for the presence of specific EBV DNA sequences by means of the polymerase chain reaction (PCR, with confirmation by direct sequencing. DESIGN AND SETTING: Prevalence study on EBV molecular genetics at a molecular pathology laboratory in a university hospital and at an applied genetics laboratory in a national institution. METHODS: A total of 75 primary glioma biopsies and 6 others from other tumors from the central nervous system were obtained. The tissues were immediately frozen for subsequent DNA extraction by means of traditional methods using proteinase K digestion and extraction with a phenol-chloroform-isoamyl alcohol mixture. DNA was precipitated with ethanol, resuspended in buffer and stored. The PCRs were carried out using primers for amplification of the EBV BamM region. Positive and negative controls were added to each reaction. The PCR products were used for direct sequencing for confirmation. RESULTS: The viral sequences were positive in 11/75 (14.7% of our samples. CONCLUSION: The prevalence of EBV DNA was 11/75 (14.7% in our glioma collection. Further molecular and epidemiological studies are needed to establish the possible role played by EBV in the tumorigenesis of gliomas.

  4. Higher prevalence of Epstein-Barr virus DNA in deeper periodontal pockets of chronic periodontitis in Japanese patients.

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    Ayako Kato

    Full Text Available Periodontitis, a complex chronic inflammatory disease caused by subgingival infection, is among the most prevalent microbial diseases in humans. Although traditional microbiological research on periodontitis has focused on putative bacteria such as Porphyromonas gingivalis, the herpes virus is proposed to be involved in the pathogenesis of periodontitis because bacterial etiology alone does not adequately explain various clinical aspects. In this study, we established for the first time, more Epstein-Barr virus (EBV DNA is found deeper in periodontal pockets of chronic periodontitis in Japanese patients. Subgingival samples were collected from 85 patients with chronic periodontitis having two periodontal sites with probing depths (PD of ≤ 3 mm (shallow or ≥ 5 mm (deep and were subjected to a nested polymerase chain reaction. EBV DNA was more frequently detected in patients with deeper PD sites (66% than in those with shallow PD sites (48% or healthy controls (45%. Coexistence of EBV DNA and P. gingivalis was significantly higher in patients with deeper PD sites (40% than in those with shallow PD sites (14% or healthy controls (13%. Although no difference in clinical index for periodontitis, the odds ratio of EBV DNA in patients with deeper PD sites was 2.36, which was 2.07-fold higher than that in those with shallow PD sites. Interestingly, the odds of acquiring chronic periodontitis (PD ≥ 5 mm were higher in the presence of both EBV DNA and P. gingivalis compared with either EBV DNA or P. gingivalis only. In addition, we also observed that EBV-encoded small RNA (EBER in positive cells of human gingival tissues. These results would suggest that EBV DNA may serve as a pathogenic factor leading to chronic periodontitis among Japanese patients.

  5. Epstein-Barr virus early antigen diffuse (EBV-EA/D)-directed immunoglobulin A antibodies in systemic lupus erythematosus patients

    DEFF Research Database (Denmark)

    Draborg, A H; Jørgensen, J M; Müller, H

    2012-01-01

    We sought to determine whether the serological response towards lytic cycle antigens of Epstein-Barr virus (EBV) is altered in systemic lupus erythematosus (SLE) patients.......We sought to determine whether the serological response towards lytic cycle antigens of Epstein-Barr virus (EBV) is altered in systemic lupus erythematosus (SLE) patients....

  6. EBV stimulates TLR- and autophagy-dependent pathways and impairs maturation in plasmacytoid dendritic cells: implications for viral immune escape.

    Science.gov (United States)

    Severa, Martina; Giacomini, Elena; Gafa, Valerie; Anastasiadou, Eleni; Rizzo, Fabiana; Corazzari, Marco; Romagnoli, Alessandra; Trivedi, Pankaj; Fimia, Gian Maria; Coccia, Eliana Marina

    2013-01-01

    Plasmacytoid DCs (pDCs) are crucial mediators in the establishment of immunity against most viruses, given their extraordinary capacity to produce a massive quantity of type I IFN. In this study we investigate the response of pDCs to infection with EBV, a γ-herpes virus that persists with an asymptomatic infection in immunocompetent hosts, although in certain conditions it can promote development of cancers or autoimmune diseases. We show that high amounts of type I IFNs were released from isolated pDCs after exposure to EBV by a mechanism requiring TLRs and a functional autophagic machinery. We next demonstrate that EBV can infect pDCs via viral binding to MHC class II molecule HLA-DR and that pDCs express EBV-induced latency genes. Furthermore, we observe that EBV is able to induce activation but not maturation of pDCs, which correlates with an impaired TNF-α release. Accordingly, EBV-infected pDCs are unable to mount a full T-cell response, suggesting that impaired pDC maturation, combined with a concomitant EBV-mediated upregulation of the T-cell inhibitory molecules B7-H1 and ICOS-L, could represent an immune-evasion strategy promoted by the virus. These mechanisms might lead to persistence in immunocompetent hosts or to dysregulated immune responses linked to EBV-associated diseases. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Sequence analysis of the Epstein-Barr virus (EBV) latent membrane protein-1 gene and promoter region

    DEFF Research Database (Denmark)

    Sandvej, K; Gratama, J W; Munch, M

    1997-01-01

    . The widespread prevalence of LMP-1 sequence variations, particularly the Xho I polymorphism and the 30-bp deletion, indicate that they cannot be used as simple markers for oncogenic viruses related to particular forms of EBV-associated tumor. Several of the structural changes detected occur, however, at sites......Sequence variations in the Epstein-Barr virus (EBV) encoded latent membrane protein-1 (LMP-1) gene have been described in a Chinese nasopharyngeal carcinoma-derived isolate (CAO), and in viral isolates from various EBV-associated tumors. It has been suggested that these genetic changes, which...... wild-type virus isolates, we sequenced the LMP-1 promoter and gene in EBV from lymphoblastoid cell lines from healthy carriers and patients without EBV-associated disease. Sequence changes were often present, and defined at least four main groups of viral isolates, which we designate Groups A through D...

  8. Prevalence of human papillomavirus and Epstein-Barr virus DNA in Chinese children with tonsillar and/or adenoidal hypertrophy.

    Science.gov (United States)

    Xue, Xiao-cheng; Chen, Xiao-ping; Yao, Wen-hao; Zhang, Yi; Sun, Guang-bin; Tan, Xue-jun

    2014-06-01

    Tonsillar and adenoidal hypertrophy are prevalent otolaryngologic disorders in children, but their pathogenesis is largely unknown. The presence of human papillomavirus (HPV) and Epstein-Barr virus (EBV) DNA in 146 tonsil and/or adenoid tissue specimens from 104 Chinese children with tonsillar and/or adenoidal hypertrophy were screened using flow-through hybridization gene-chip technology and real-time fluorescence-based quantitative PCR. Then, the relationships between the prevalence of the viruses and other clinical characteristics of tonsillar and/or adenoidal hypertrophy were analyzed. No patient had HPV DNA. EBV DNA was detected in 19/42 (45.2%) tonsil tissues and 72/104 (69.2%) adenoid tissue specimens (P hypertrophy. Adenoid tissues might more susceptible than tonsil tissues to EBV infection. In addition, EBV infection did not aggravate snoring in patients with tonsillar and/or adenoidal hypertrophy. © 2014 Wiley Periodicals, Inc.

  9. P-glycoprotein is expressed and causes resistance to chemotherapy in EBV-positive T-cell lymphoproliferative diseases

    International Nuclear Information System (INIS)

    Yoshimori, Mayumi; Takada, Honami; Imadome, Ken-Ichi; Kurata, Morito; Yamamoto, Kouhei; Koyama, Takatoshi; Shimizu, Norio; Fujiwara, Shigeyoshi; Miura, Osamu; Arai, Ayako

    2015-01-01

    Epstein–Barr virus-positive T-cell lymphoproliferative diseases (EBV-T-LPDs) are rare lymphomas with poor prognosis. Although chemotherapeutic strategies such as CHOP have been often selected, they have exhibited only limited efficacy. To clarify the mechanism of chemoresistance, we examined P-glycoprotein (P-gp) expression. P-gp acts as an energy-dependent efflux pump that excretes drugs from the cytoplasm, resulting in low-intracellular drug concentrations and poor sensitivity to chemotherapy. We examined P-gp expression in EBV-positive cells by immunohistochemistry staining in three patients of EBV-T-LPDs and the expression was detected in all patients. We also examined mdr1 mRNA expression by reverse-transcriptase polymerase-chain reaction (RT-PCR) in EBV-positive tumor cells from these patients and additional three patients. The expression was detected in all examined patients. In five EBV-T-LPDs patients, P-gp function was detected by Rhodamine-123 efflux assay in these cells. The efflux was inhibited by treatment with a P-gp inhibitor, cyclosporine A (CsA). We also examined and detected P-gp expression in EBV-positive T-cell lines SNT8 and SNT16 established from EBV-T-LPDs patients, by RT-PCR and western blotting. The function was also detected by Rhodamine-123 efflux in these cell lines. Inhibition and knock down of P-gp by CsA and siRNA, respectively, enhanced etoposide- and doxorubicin-induced cell death in the EBV-positive T-cell lines. Finally, we infected the T-cell line MOLT4 with EBV, and found that mdr1 mRNA expression and Rhodamine 123 efflux were upregulated after infection. These results indicated that enhanced P-gp expression contributed to the chemoresistance of EBV-T-LPDs

  10. EBV-positive diffuse large B-cell lymphoma in young adults: is this a distinct disease entity?

    Science.gov (United States)

    Hong, J Y; Yoon, D H; Suh, C; Huh, J; Do, I-G; Sohn, I; Jo, J; Jung, S-H; Hong, M E; Yoon, H; Ko, Y H; Kim, S J; Kim, W S

    2015-03-01

    Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) of the elderly is defined only in adults older than 50 years. However, EBV-positive DLBCL can affect younger patients. We investigated the prevalence, clinical characteristics and survival outcomes of EBV-positive DLBCL in young adults. We analyzed patients with de novo DLBCL who were registered in the Samsung Medical Center (SMC) retrospective lymphoma cohort and prospective SMC Lymphoma Cohort Study I (ClinicalTrials.gov: NCT00822731). A total of 571 cases were included in the analysis. The prevalence of EBV positivity was 6.7% (13/195) and 9.3% (35/376) in the young group (≤50 years) and in the elderly group (>50 years), respectively. EBV status was closely associated with unique unfavorable clinical characteristics [older age, more advanced stage, two or more sites of extranodal involvement, higher International Prognostic Index (IPI), and age-adjusted IPI risk] only in the elderly group. Poor prognostic impact of EBV positivity on overall survival was observed only in the elderly group [hazard ratio (HR) 2.86; 95% confidence interval (CI) 1.83-4.47; P young group (HR 1.17; 95% CI 0.35-3.89; P = 0.801). A substantial proportion of EBV-positive DLBCL of the elderly can occur in young adults. EBV positivity of DLBCL in young adults was not associated with unfavorable clinical characteristics or worse outcomes. We suggest that EBV-positive DLBCL should not be confined only in the elderly and 'EBV-positive DLBCL in young adults' needs to be considered as a clinically distinct disease entity. ClinicalTrials.gov: NCT02060435. © The Author 2014. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  11. Methods for pretreating biomass

    Science.gov (United States)

    Balan, Venkatesh; Dale, Bruce E; Chundawat, Shishir; Sousa, Leonardo

    2017-05-09

    A method for pretreating biomass is provided, which includes, in a reactor, allowing gaseous ammonia to condense on the biomass and react with water present in the biomass to produce pretreated biomass, wherein reactivity of polysaccharides in the biomass is increased during subsequent biological conversion as compared to the reactivity of polysaccharides in biomass which has not been pretreated. A method for pretreating biomass with a liquid ammonia and recovering the liquid ammonia is also provided. Related systems which include a biochemical or biofuel production facility are also disclosed.

  12. Progesterone analogues reduce plasma Epstein–Barr virus DNA load and improve pain control in recurrent/metastatic nasopharyngeal carcinoma patients under supportive care

    Directory of Open Access Journals (Sweden)

    Chia-Yen Hung

    2017-08-01

    Conclusion: Progesterone analogues improve quality of life in terms of performance status, pain control, and plasma EBV DNA load in patients with locally recurrent/metastatic NPC under palliative care.

  13. Towards global interoperability for supporting biodiversity research on Essential Biodiversity Variables (EBVs)

    NARCIS (Netherlands)

    Kissling, W.D.; Hardisty, A.; García, E.A.; Santamaria, M.; De Leo, F.; Pesole, G.; Freyhof, J.; Manset, D.; Wissel, S.; Konijn, J.; Los, W.

    2015-01-01

    Essential biodiversity variables (EBVs) have been proposed by the Group on Earth Observations Biodiversity Observation Network (GEO BON) to identify a minimum set of essential measurements that are required for studying, monitoring and reporting biodiversity and ecosystem change. Despite the initial

  14. Pretreatment of microbial sludges

    Science.gov (United States)

    Rivard, Christopher J.; Nagle, Nicholas J.

    1995-01-01

    Methods are described for pretreating microbial sludges to break cells and disrupt organic matter. One method involves the use of sonication, and another method involves the use of shear forces. The pretreatment of sludge enhances bioconversion of the organic fraction. This allows for efficient dewatering of the sludge and reduces the cost for final disposal of the waste.

  15. Histone deacetylase inhibitors are potent inducers of gene expression in latent EBV and sensitize lymphoma cells to nucleoside antiviral agents.

    Science.gov (United States)

    Ghosh, Sajal K; Perrine, Susan P; Williams, Robert M; Faller, Douglas V

    2012-01-26

    Induction of EBV lytic-phase gene expression, combined with exposure to an antiherpes viral drug, represents a promising targeted therapeutic approach to EBV-associated lymphomas. Short-chain fatty acids or certain chemotherapeutics have been used to induce EBV lytic-phase gene expression in cultured cells and mouse models, but these studies generally have not translated into clinical application. The recent success of a clinical trial with the pan-histone deacetylase (pan-HDAC) inhibitor arginine butyrate and the antiherpes viral drug ganciclovir in the treatment of EBV lymphomas prompted us to investigate the potential of several HDAC inhibitors, including some new, highly potent compounds, to sensitize EBV(+) human lymphoma cells to antiviral agents in vitro. Our study included short-chain fatty acids (sodium butyrate and valproic acid); hydroxamic acids (oxamflatin, Scriptaid, suberoyl anilide hydroxamic acid, panobinostat [LBH589], and belinostat [PXD101]); the benzamide MS275; the cyclic tetrapeptide apicidin; and the recently discovered HDAC inhibitor largazole. With the exception of suberoyl anilide hydroxamic acid and PXD101, all of the other HDAC inhibitors effectively sensitized EBV(+) lymphoma cells to ganciclovir. LBH589, MS275, and largazole were effective at nanomolar concentrations and were 10(4) to 10(5) times more potent than butyrate. The effectiveness and potency of these HDAC inhibitors make them potentially applicable as sensitizers to antivirals for the treatment of EBV-associated lymphomas.

  16. Efficacy of the ND:YAG laser therapy on EBV and HSV1 contamination in periodontal pockets.

    Science.gov (United States)

    Martelli, Francesco Saverio; Bacci, Giovanni; Martelli, Maria Laura; Nobili, Piero; Boddi, Anna; Rosati, Claudio; Fanti, Elena

    2015-01-01

    The aim of this retrospective multicenter study was to verify the efficacy of Nd:YAG laser in the treatment of periodontal pockets infected by Epstein-Barr Virus (EBV) and Herpes Simplex Virus 1 (HSV1). Subgingival plaque samples of 291 Italian periodontal patients were analyzed by Real Time PCR to evaluate the frequency of both viruses before and after Nd:YAG laser-assisted periodontal treatment. Before treatment, EBV and HSV1 were observed in 29.9% and in 3.8% of periodontal patients respectively, while co-infection with both viruses was detected in 1.7% of cases. Periodontal Nd:YAG laser treatment ("Periodontal Biological Laser-Assisted Therapy", PERIOBLAST) produced statistical significant benefits, especially in EBV periodontal infection: 78.2% of EBV positive patients became EBV-negative following treatment. Results of this preliminary study highlight that EBV is found in periodontal pockets more frequently than HSV1, supporting the theory of the potential role of EBV in the onset and progression of periodontal disease. Moreover, our data showed that Nd:YAG laser-assisted periodontal treatment (Perioblast) is also effective in case of viral infection, validating evidences that it represents a successful alternative approach to traditional periodontal protocols.

  17. Epstein-Barr virus early antigen diffuse (EBV-EA/D)-directed immunoglobulin A antibodies in systemic lupus erythematosus patients.

    Science.gov (United States)

    Draborg, A H; Jørgensen, J M; Müller, H; Nielsen, C T; Jacobsen, S; Iversen, L V; Theander, E; Nielsen, L P; Houen, G; Duus, K

    2012-08-01

    We sought to determine whether the serological response towards lytic cycle antigens of Epstein-Barr virus (EBV) is altered in systemic lupus erythematosus (SLE) patients. We used enzyme-linked immunosorbent assay (ELISA) to investigate the prevalence of EBV early antigen diffuse (EBV-EA/D) antibodies in sera from 60 patients with SLE, 40 with scleroderma (SSc), 20 with primary Sjögren's syndrome (pSS), 20 with rheumatoid arthritis (RA), 20 healthy controls, and also subjects with various circulating autoantibodies. Samples from patients were obtained from clinics specialized within the diseases in Denmark and Sweden and samples from healthy controls were obtained from volunteers. A significant elevated titre of immunoglobulin (Ig)A, IgG, and IgM EBV-EA/D antibodies was found in SLE patients compared to healthy controls, a finding not explained by immunosuppressive treatment or disease activity. The largest difference was observed for IgA EBV-EA/D antibodies (p = 0.0013) with a seropositive rate of 58% in SLE patients and 0% in healthy controls. RA and SSc patients and individuals seropositive for anti-Scl-70 were additionally found to have elevated titres of IgA EBV-EA/D antibodies (40%, p = 0.014; 60%, p = 0.015; and 38.5%, p = 0.045, respectively). However, the titres were generally lower than in SLE patients. Our findings support an association between EBV and SLE. The elevated titre of EBV-EA/D-directed IgA antibodies found in SLE patients could suggest reactivation of EBV in epithelial cells or reinfection of epithelial cells after reactivation in B cells, indicating lack of control of the latent infection.

  18. Latent Membrane Protein LMP2A Impairs Recognition of EBV-Infected Cells by CD8+ T Cells.

    Directory of Open Access Journals (Sweden)

    Chiara Rancan

    2015-06-01

    Full Text Available The common pathogen Epstein-Barr virus (EBV transforms normal human B cells and can cause cancer. Latent membrane protein 2A (LMP2A of EBV supports activation and proliferation of infected B cells and is expressed in many types of EBV-associated cancer. It is not clear how latent EBV infection and cancer escape elimination by host immunity, and it is unknown whether LMP2A can influence the interaction of EBV-infected cells with the immune system. We infected primary B cells with EBV deleted for LMP2A, and established lymphoblastoid cell lines (LCLs. We found that CD8+ T cell clones showed higher reactivity against LMP2A-deficient LCLs compared to LCLs infected with complete EBV. We identified several potential mediators of this immunomodulatory effect. In the absence of LMP2A, expression of some EBV latent antigens was elevated, and cell surface expression of MHC class I was marginally increased. LMP2A-deficient LCLs produced lower amounts of IL-10, although this did not directly affect CD8+ T cell recognition. Deletion of LMP2A led to several changes in the cell surface immunophenotype of LCLs. Specifically, the agonistic NKG2D ligands MICA and ULBP4 were increased. Blocking experiments showed that NKG2D activation contributed to LCL recognition by CD8+ T cell clones. Our results demonstrate that LMP2A reduces the reactivity of CD8+ T cells against EBV-infected cells, and we identify several relevant mechanisms.

  19. Prevalence of human papillomavirus and Epstein-Barr virus DNA in penile cancer cases from Brazil

    Directory of Open Access Journals (Sweden)

    Larissa Alves Afonso

    2012-02-01

    Full Text Available Penile cancer is a potentially mutilating disease. Although its occurrence is relatively rare worldwide, penile cancer rates can be high in developing countries. A few studies have been conducted on the involvement of human papillomavirus (HPV in penile carcinoma, which have found HPV present in 30-70% of penile malignant lesions, with a higher prevalence of HPV 16 and 18. It has been assumed that cofactors, such as Epstein-Barr virus (EBV infections, may play a role in the progression of penile neoplasia. The aim of this study was to determine HPV and EBV prevalence in 135 penile malignant lesions from Brazilian men through the use of MY09/11 polymerase chain reaction (PCR, type-specific PCR and restriction fragment length polymorphism analysis. HPV prevalence among the men tested was 60.7%. Of the men who tested positive, 27 presented with HPV 16 (29.7%, five with HPV 18 (5.5%, 21 with HPV 45 (23.1% and nine with HPV 6 (9.9%. Seven mixed infections were detected (9.2%, while 11 cases remained untyped (13.4%. Regarding EBV positivity, 46.7% of the samples contained EBV DNA with EBV-1 as the most prevalent type (74.6%. More than 23% of the men were co-infected with both HPV and EBV, while 35% presented exclusively with HPV DNA and 20% presented only with EBV DNA. Penile carcinoma aetiology has not been fully elucidated and the role of HPV and EBV infections individually or synergistically is still controversial. Hence, more studies are needed to determine their possible role in carcinogenesis.

  20. Quantitative detection of epstein-barr virus DNA in cerebrospinal fluid and blood samples of patients with relapsing-remitting multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Clementina E Cocuzza

    Full Text Available The presence of Epstein-Barr Virus (EBV DNA in cerebrospinal fluid (CSF and peripheral blood (PB samples collected from 55 patients with clinical and radiologically-active relapsing-remitting MS (RRMS and 51 subjects with other neurological diseases was determined using standardized commercially available kits for viral nucleic acid extraction and quantitative EBV DNA detection. Both cell-free and cell-associated CSF and PB fractions were analyzed, to distinguish latent from lytic EBV infection. EBV DNA was detected in 5.5% and 18.2% of cell-free and cell-associated CSF fractions of patients with RRMS as compared to 7.8% and 7.8% of controls; plasma and peripheral blood mononuclear cells (PBMC positivity rates were 7.3% and 47.3% versus 5.8% and 31.4%, respectively. No significant difference in median EBV viral loads of positive samples was found between RRMS and control patients in all tested samples. Absence of statistically significant differences in EBV positivity rates between RRMS and control patients, despite the use of highly sensitive standardized methods, points to the lack of association between EBV and MS disease activity.

  1. Plasma Epstein-Barr virus DNA for pediatric Burkitt lymphoma diagnosis, prognosis and response assessment in Malawi.

    Science.gov (United States)

    Westmoreland, Katherine D; Montgomery, Nathan D; Stanley, Christopher C; El-Mallawany, Nader Kim; Wasswa, Peter; van der Gronde, Toon; Mtete, Idah; Butia, Mercy; Itimu, Salama; Chasela, Mary; Mtunda, Mary; Kampani, Coxcilly; Liomba, N George; Tomoka, Tamiwe; Dhungel, Bal M; Sanders, Marcia K; Krysiak, Robert; Kazembe, Peter; Dittmer, Dirk P; Fedoriw, Yuri; Gopal, Satish

    2017-06-01

    Point-of-care tools are needed in sub-Saharan Africa (SSA) to improve pediatric Burkitt lymphoma (BL) diagnosis and treatment. We evaluated plasma Epstein-Barr virus (pEBV) DNA as a pediatric BL biomarker in Malawi. Prospectively enrolled children with BL were compared to classical Hodgkin lymphoma (cHL) and nonlymphoma diagnoses. Pediatric BL patients received standardized chemotherapy and supportive care. pEBV DNA was measured at baseline, mid-treatment, and treatment completion. Of 121 assessed children, pEBV DNA was detected in 76/88 (86%) with BL, 16/17 (94%) with cHL, and 2/16 (12%) with nonlymphoma, with proportions higher in BL versus nonlymphoma (p < 0.001) and similar in BL versus cHL (p = 0.69). If detected, median pEBV DNA was 6.1 log 10 copies/mL for BL, 4.8 log 10 copies/mL for cHL, and 3.4 log 10 copies/mL for nonlymphoma, with higher levels in BL versus cHL (p = 0.029), and a trend toward higher levels in BL versus nonlymphoma (p = 0.062). pEBV DNA declined during treatment in the cohort overall and increased in several children before clinical relapse. Twelve-month overall survival was 40% in the cohort overall, and for children with baseline pEBV detected, survival was worse if baseline pEBV DNA was ≥6 log 10 copies/mL versus <6 log 10 copies/mL (p = 0.0002), and also if pEBV DNA was persistently detectable at mid-treatment versus undetectable (p = 0.041). Among children with baseline pEBV DNA detected, viremia was the only significant risk factor for death by 12 months in multivariate analyses (adjusted hazard ratio 1.35 per log 10 copies/mL, 95% CI 1.04-1.75, p = 0.023). Quantitative pEBV DNA has potential utility for diagnosis, prognosis, and response assessment for pediatric BL in SSA. © 2017 UICC.

  2. Uracil DNA glycosylase BKRF3 contributes to Epstein-Barr virus DNA replication through physical interactions with proteins in viral DNA replication complex.

    Science.gov (United States)

    Su, Mei-Tzu; Liu, I-Hua; Wu, Chia-Wei; Chang, Shu-Ming; Tsai, Ching-Hwa; Yang, Pei-Wen; Chuang, Yu-Chia; Lee, Chung-Pei; Chen, Mei-Ru

    2014-08-01

    Epstein-Barr virus (EBV) BKRF3 shares sequence homology with members of the uracil-N-glycosylase (UNG) protein family and has DNA glycosylase activity. Here, we explored how BKRF3 participates in the DNA replication complex and contributes to viral DNA replication. Exogenously expressed Flag-BKRF3 was distributed mostly in the cytoplasm, whereas BKRF3 was translocated into the nucleus and colocalized with the EBV DNA polymerase BALF5 in the replication compartment during EBV lytic replication. The expression level of BKRF3 increased gradually during viral replication, coupled with a decrease of cellular UNG2, suggesting BKRF3 enzyme activity compensates for UNG2 and ensures the fidelity of viral DNA replication. In immunoprecipitation-Western blotting, BKRF3 was coimmuno-precipitated with BALF5, the polymerase processivity factor BMRF1, and the immediate-early transactivator Rta. Coexpression of BMRF1 appeared to facilitate the nuclear targeting of BKRF3 in immunofluorescence staining. Residues 164 to 255 of BKRF3 were required for interaction with Rta and BALF5, whereas residues 81 to 166 of BKRF3 were critical for BMRF1 interaction in glutathione S-transferase (GST) pulldown experiments. Viral DNA replication was defective in cells harboring BKRF3 knockout EBV bacmids. In complementation assays, the catalytic mutant BKRF3(Q90L,D91N) restored viral DNA replication, whereas the leucine loop mutant BKRF3(H213L) only partially rescued viral DNA replication, coupled with a reduced ability to interact with the viral DNA polymerase and Rta. Our data suggest that BKRF3 plays a critical role in viral DNA synthesis predominantly through its interactions with viral proteins in the DNA replication compartment, while its enzymatic activity may be supplementary for uracil DNA glycosylase (UDG) function during virus replication. Catalytic activities of both cellular UDG UNG2 and viral UDGs contribute to herpesviral DNA replication. To ensure that the enzyme activity executes at

  3. Molecular Pathogenesis of EBV Susceptibility in XLP as Revealed by Analysis of Female Carriers with Heterozygous Expression of SAP

    Science.gov (United States)

    Palendira, Umaimainthan; Low, Carol; Chan, Anna; Hislop, Andrew D.; Ho, Edwin; Phan, Tri Giang; Deenick, Elissa; Cook, Matthew C.; Riminton, D. Sean; Choo, Sharon; Loh, Richard; Alvaro, Frank; Booth, Claire; Gaspar, H. Bobby; Moretta, Alessandro; Khanna, Rajiv; Rickinson, Alan B.; Tangye, Stuart G.

    2011-01-01

    X-linked lymphoproliferative disease (XLP) is a primary immunodeficiency caused by mutations in SH2D1A which encodes SAP. SAP functions in signalling pathways elicited by the SLAM family of leukocyte receptors. A defining feature of XLP is exquisite sensitivity to infection with EBV, a B-lymphotropic virus, but not other viruses. Although previous studies have identified defects in lymphocytes from XLP patients, the unique role of SAP in controlling EBV infection remains unresolved. We describe a novel approach to this question using female XLP carriers who, due to random X-inactivation, contain both SAP+ and SAP− cells. This represents the human equivalent of a mixed bone marrow chimera in mice. While memory CD8+ T cells specific for CMV and influenza were distributed across SAP+ and SAP− populations, EBV-specific cells were exclusively SAP+. The preferential recruitment of SAP+ cells by EBV reflected the tropism of EBV for B cells, and the requirement for SAP expression in CD8+ T cells for them to respond to Ag-presentation by B cells, but not other cell types. The inability of SAP− clones to respond to Ag-presenting B cells was overcome by blocking the SLAM receptors NTB-A and 2B4, while ectopic expression of NTB-A on fibroblasts inhibited cytotoxicity of SAP− CD8+ T cells, thereby demonstrating that SLAM receptors acquire inhibitory function in the absence of SAP. The innovative XLP carrier model allowed us to unravel the mechanisms underlying the unique susceptibility of XLP patients to EBV infection in the absence of a relevant animal model. We found that this reflected the nature of the Ag-presenting cell, rather than EBV itself. Our data also identified a pathological signalling pathway that could be targeted to treat patients with severe EBV infection. This system may allow the study of other human diseases where heterozygous gene expression from random X-chromosome inactivation can be exploited. PMID:22069374

  4. Molecular pathogenesis of EBV susceptibility in XLP as revealed by analysis of female carriers with heterozygous expression of SAP.

    Directory of Open Access Journals (Sweden)

    Umaimainthan Palendira

    2011-11-01

    Full Text Available X-linked lymphoproliferative disease (XLP is a primary immunodeficiency caused by mutations in SH2D1A which encodes SAP. SAP functions in signalling pathways elicited by the SLAM family of leukocyte receptors. A defining feature of XLP is exquisite sensitivity to infection with EBV, a B-lymphotropic virus, but not other viruses. Although previous studies have identified defects in lymphocytes from XLP patients, the unique role of SAP in controlling EBV infection remains unresolved. We describe a novel approach to this question using female XLP carriers who, due to random X-inactivation, contain both SAP(+ and SAP(- cells. This represents the human equivalent of a mixed bone marrow chimera in mice. While memory CD8(+ T cells specific for CMV and influenza were distributed across SAP(+ and SAP(- populations, EBV-specific cells were exclusively SAP(+. The preferential recruitment of SAP(+ cells by EBV reflected the tropism of EBV for B cells, and the requirement for SAP expression in CD8(+ T cells for them to respond to Ag-presentation by B cells, but not other cell types. The inability of SAP(- clones to respond to Ag-presenting B cells was overcome by blocking the SLAM receptors NTB-A and 2B4, while ectopic expression of NTB-A on fibroblasts inhibited cytotoxicity of SAP(- CD8(+ T cells, thereby demonstrating that SLAM receptors acquire inhibitory function in the absence of SAP. The innovative XLP carrier model allowed us to unravel the mechanisms underlying the unique susceptibility of XLP patients to EBV infection in the absence of a relevant animal model. We found that this reflected the nature of the Ag-presenting cell, rather than EBV itself. Our data also identified a pathological signalling pathway that could be targeted to treat patients with severe EBV infection. This system may allow the study of other human diseases where heterozygous gene expression from random X-chromosome inactivation can be exploited.

  5. The Relationship Between N(Htotal) and E(B-V) at High Galactic Latitude

    Science.gov (United States)

    Doppel, Jessica Erin; Magnani, Loris A.

    2017-06-01

    We studied the gas-to-dust ratio at high Galactic latitudes (|b| ≥ 20° and 30°). The gas content was measured by the column density of atomic hydrogen from the Leiden/Argentine/Bonn survey and the dust by E(B-V) from the Schlafly et al. (2014) compilation. We do not see significant variation as a function of galactic quadrant or hemisphere. Our results are similar to other studies that used different techniques for deriving N(HI), N(H2), or E(B-V). Like Liszt (2013), we see a higher slope [N(HI)/E(B-V)] if we choose only those points with E(B-V) ≤ 0.1 mag. We also see the break in slope he noted at 0.08 mag. We examined lines of sight that coincided with the Georgia/Harvard Smithsonian CfA high-latitude CO(1-0) surveys. Although at |b| ≥ 30° there were only about 200 CO detections, those points lie systematically below our best fit lines to the data. We can convert the CO line intensities to N(H2) and determine a global CO-H2 conversion factor for the high latitude sky.

  6. Triterpenoids from Ganoderma lucidum inhibit the activation of EBV antigens as telomerase inhibitors.

    Science.gov (United States)

    Zheng, Dong-Shu; Chen, Liang-Shu

    2017-10-01

    Nasopharyngeal carcinoma (NPC) is a malignant disease that threatens the health of humans. To find effective agents for the inhibition of Epstein-Barr virus (EBV) infection, which is associated with NPC, a phytochemical investigation of Ganoderma lucidum was carried out in the present study. Five triterpenoids were identified, including ganoderic acid A (compound 1), ganoderic acid B (compound 2), ganoderol B (compound 3), ganodermanontriol (compound 4), and ganodermanondiol (compound 5), on the basis of spectroscopic analysis. An inhibition of EBV antigens activation assay was implemented to elucidate the triterpenoids from G. lucidum and potentially prevent NPC. All the triterpenoids showed significant inhibitory effects on both EBV EA and CA activation at 16 nmol. At 3.2 nmol, all the compounds moderately inhibited the activation of the two antigens. The activity of telomerase was inhibited by these triterpenoids at 10 µM. Molecular docking demonstrated that compound 1 was able to inhibit telomerase as a ligand. In addition, the physicochemical properties of these compounds were calculated to elucidate their drug-like properties. These results provided evidence for the application of these triterpenoids and whole G. lucidum in the treatment of NPC.

  7. A novel immune evasion mechanism of LMP-1, an EBV-primary oncogene, in nasopharyngeal carcinoma.

    Science.gov (United States)

    Yoshizaki, Tomokazu

    2011-01-01

    Nasopharyngeal carcinoma is an Epstein-Barr virus (EBV)-associated tumor. Viruses that are associated with malignant transformation have evolved unique mechanisms to interfere with this interaction to evade antiviral T cell responses. EBV exploits many immune evasive strategies to successfully establish a latent infection in B cells. CD8+ T cell responses to LMP-1 are generally very low and rarely detected in healthy virus carriers. Activation of the NF-kB pathway by EBV-LMP-1 leads to an upregulation of the MHC class I antigen-processing pathway. Paradoxically, LMP-1itself induces a subdominant CD8+ T cell response and appears to have evolved to avoid immune recognition. An expression of LMP-1 in human cells enhanced the trans-presentation of CD8+ T cell epitopes; however, cis-presentation of LMP-1-derived epitopes was severely impaired. Deletion of the first transmembrane domain of LMP-1, which prevented self-aggregation, significantly enhanced the cis-presentation of T cell epitopes from this protein, whereas it lost its ability to upregulate trans-presentation. These results delineate a novel mechanism of immune evasion, which renders a virally encoded oncogene inaccessible to the conventional MHC class I pathway limiting its cis-presentation. Copyright © 2011 S. Karger AG, Basel.

  8. GREET Pretreatment Module

    Energy Technology Data Exchange (ETDEWEB)

    Adom, Felix K. [Argonne National Lab. (ANL), Argonne, IL (United States). Energy Systems Division; Dunn, Jennifer B. [Argonne National Lab. (ANL), Argonne, IL (United States). Energy Systems Division; Han, Jeongwoo [Argonne National Lab. (ANL), Argonne, IL (United States). Energy Systems Division

    2014-09-01

    A wide range of biofuels and biochemicals can be produced from cellulosic biomass via different pretreatment technologies that yield sugars. Process simulations of dilute acid and ammonia fiber expansion pretreatment processes and subsequent hydrolysis were developed in Aspen Plus for four lignocellulosic feedstocks (corn stover, miscanthus, switchgrass, and poplar). This processing yields sugars that can be subsequently converted to biofuels or biochemical. Material and energy consumption data from Aspen Plus were then compiled in a new Greenhouses Gases, Regulated Emissions, and Energy Use in Transportation (GREETTM) pretreatment module. The module estimates the cradle-to-gate fossil energy consumption (FEC) and greenhouse gas (GHG) emissions associated with producing fermentable sugars. This report documents the data and methodology used to develop this module and the cradle-to-gate FEC and GHG emissions that result from producing fermentable sugars.

  9. Use of Foscarnet Therapy for EBV Infection following Control of PTLD with Enhancement of Cellular Immunity in a Lung-Transplant Recipient

    Directory of Open Access Journals (Sweden)

    Kamyar Afshar

    2011-01-01

    Full Text Available Posttransplant lymphoproliferative disorder (PTLD is a serious complication following solid organ transplantation with an annual incidence rate of 3–5% in lung-transplant recipients. Pathogenesis indicates a strong association with functional over-immunosuppression and EBV infection. Clinical improvement is generally observed with reduction in immunosuppression intensity alone. We present a case of a 24-year-old woman with EBV-associated PTLD following lung transplant where decreasing the immunosuppression improved PTLD but was ineffective against controlling the EBV infection. Foscarnet in combination with immunoglobulins was successfully administered to cause a remission of the EBV infection. This is the second case reported of a persistent EBV infection after reducing immunosuppression levels and evidence of PTLD remission that required foscarnet for EBV infection control.

  10. KSHV LANA and EBV LMP1 induce the expression of UCH-L1 following viral transformation

    Energy Technology Data Exchange (ETDEWEB)

    Bentz, Gretchen L.; Bheda-Malge, Anjali; Wang, Ling [Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill (United States); Shackelford, Julia [Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill (United States); Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill (United States); Damania, Blossom [Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill (United States); Departments of Medicine and of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC (United States); Pagano, Joseph S., E-mail: joseph_pagano@med.unc.edu [Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill (United States); Departments of Medicine and of Microbiology and Immunology, University of North Carolina, Chapel Hill, NC (United States)

    2014-01-05

    Ubiquitin C-terminal Hydrolase L1 (UCH-L1) has oncogenic properties and is highly expressed during malignancies. We recently documented that Epstein-Barr virus (EBV) infection induces uch-l1 expression. Here we show that Kaposi's Sarcoma-associated herpesvirus (KSHV) infection induced UCH-L1 expression, via cooperation of KSHV Latency-Associated Nuclear Antigen (LANA) and RBP-Jκ and activation of the uch-l1 promoter. UCH-L1 expression was also increased in Primary Effusion Lymphoma (PEL) cells co-infected with KSHV and EBV compared with PEL cells infected only with KSHV, suggesting EBV augments the effect of LANA on uch-l1. EBV latent membrane protein 1 (LMP1) is one of the few EBV products expressed in PEL cells. Results showed that LMP1 was sufficient to induce uch-l1 expression, and co-expression of LMP1 and LANA had an additive effect on uch-l1 expression. These results indicate that viral latency products of both human γ-herpesviruses contribute to uch-l1 expression, which may contribute to the progression of lymphoid malignancies. - Highlights: • Infection of endothelial cells with KSHV induced UCH-L1 expression. • KSHV LANA is sufficient for the induction of uch-l1. • Co-infection with KSHV and EBV (observed in some PELs) results in the additive induction of uch-l1. • EBV LMP1 also induced UCH-L1 expression. • LANA- and LMP1-mediated activation of the uch-l1 promoter is in part through RBP-Jκ.

  11. Using Essential Biodiversity Variables (EBVs) As a Framework for Coordination Between Research and Monitoring Networks: A Case Study with Phenology

    Science.gov (United States)

    Weltzin, J. F.; Jones, K. D.; Brown, J. F.; Elmendorf, S.; Enquist, C.; Rosemartin, A.; Thorpe, A.; Wee, B.

    2014-12-01

    The United Nations Convention on Biological Diversity (CBD) was organized to encourage countries to take action to address issues of declining biodiversity. In2010, the CBD identified specific goals for 2011-2020 (the "Aichi Targets") and a tiered system of indicators necessary to achieve those targets. Essential biodiversity variables (EBVs) are the standardized measurements and observations at the base of this system; they are the basic level of information that is necessary to calculate these indicators. By providing a list of pre-defined EBVs, existing research and research planned for the future can align measurements to address common questions. We assessed the applicability of phenology EBVs for standardizing measurements across observation networks within the US as a test case for use of the standardized used of EBVs. Phenology products from the USA National Phenology Network, a citizen science observer based program, NEON, a multi-scale ecological observatory, and remotely sensed data from USGS EROS were considered for this purpose. Essential Biodiversity Variables currently defined for phenology are insufficient to support consistent measurement across monitoring networks. Specifically, phenology which is a field of study, is currently listed as a single EBV within the general category of 'species traits'. With the only guidance provided to future observation networks being that of measuring 'phenology,' there would likely be as many approaches to achieving this goal as networks participating. We propose more narrowly defined variables which may be more appropriate for standardization and demonstrate how these measurements satisfy the basic characteristics of an EBV in that they are relevant, sensitive to change, biological and generalizable, scalable, feasible, stable and, represent state variables. We map these variables to the tiered indicators identified by the CBD and, finally, to Aichi Targets to which they contribute. EBVs may be used not only to

  12. B cell differentiation in EBV-positive Burkitt Lymphoma is impaired at post-transcriptional level by miRNA altered expression

    DEFF Research Database (Denmark)

    Leucci, E; Onnis, A; Cocco, M

    2009-01-01

    investigated the expression of specific miRNAs predicted to be involved in B cell differentiation and we found that hsa-miR-127 is differentially expressed between EBV-positive and EBV-negative BLs. In particular, it was strongly up-regulated only in EBV-positive BL samples, whereas EBV-negative cases showed...... levels of expression similar to normal controls, including microdissected GC cells.In addition, we found evidence that hsa-miR-127 is involved in B cell differentiation process through post transcriptional regulation of BLIMP1 and XBP1. The over-expression of this miRNA may thus represent a key event...

  13. Plasticity of DNA replication initiation in Epstein-Barr virus episomes.

    Directory of Open Access Journals (Sweden)

    Paolo Norio

    2004-06-01

    Full Text Available In mammalian cells, the activity of the sites of initiation of DNA replication appears to be influenced epigenetically, but this regulation is not fully understood. Most studies of DNA replication have focused on the activity of individual initiation sites, making it difficult to evaluate the impact of changes in initiation activity on the replication of entire genomic loci. Here, we used single molecule analysis of replicated DNA (SMARD to study the latent duplication of Epstein-Barr virus (EBV episomes in human cell lines. We found that initiation sites are present throughout the EBV genome and that their utilization is not conserved in different EBV strains. In addition, SMARD shows that modifications in the utilization of multiple initiation sites occur across large genomic regions (tens of kilobases in size. These observations indicate that individual initiation sites play a limited role in determining the replication dynamics of the EBV genome. Long-range mechanisms and the genomic context appear to play much more important roles, affecting the frequency of utilization and the order of activation of multiple initiation sites. Finally, these results confirm that initiation sites are extremely redundant elements of the EBV genome. We propose that these conclusions also apply to mammalian chromosomes.

  14. Advances in aluminum pretreatment

    Energy Technology Data Exchange (ETDEWEB)

    Sudour, Michel; Maintier, Philippe [PPG Industries France, 3 Z.A.E. Les Dix Muids, B.P. 89, F-59583 Marly (France); Simpson, Mark [PPG Industries Inc., 1200 Piedmont Troy, Michigan 48083 (United States); Quaglia, Paolo [PPG Industries Italia, Via Garavelli 21, I-15028 Quattordio (Italy)

    2004-07-01

    As automotive manufacturers continue to look for ways to reduce vehicle weight, aluminum is finding more utility as a body panel component. The substitution of cold-rolled steel and zinc-coated substrates with aluminum has led to new challenges in vehicle pretreatment. As a result, changes to traditional pretreatment chemistries and operating practices are necessary in order to produce an acceptable coating on aluminum body panels. These changes result in increased sludging and other undesirable characteristics. In addition to the chemistry changes, there are also process-related problems to consider. Many existing automotive pretreatment lines simply were not designed to handle aluminum and its increased demands on filtration and circulation equipment. To retrofit such a system is capital intensive and in addition to requiring a significant amount of downtime, may not be totally effective. Thus, the complexities of pre-treating aluminum body panels have actually had a negative effect on efforts to introduce more aluminum into new vehicle design programs. Recent research into ways of reducing the negative effects has led to a new understanding of the nature of zinc phosphate bath -aluminum interactions. Many of the issues associated with the pretreatment of aluminum have been identified and can be mitigated with only minor changes to the zinc phosphate bath chemistry. The use of low levels of soluble Fe ions, together with free fluoride, has been shown to dramatically improve the efficiency of a zinc phosphate system processing aluminum. Appearance of zinc phosphate coatings, coating weights and sludge are all benefited by this chemistry change. (authors)

  15. Encephalitis treatment – a case report with long-term follow-up of EBV PCR in cerebrospinal fluid

    Directory of Open Access Journals (Sweden)

    Zarlasht F

    2017-10-01

    Full Text Available Fnu Zarlasht,1 Mashal Salehi,2 Mohammad Abu-Hishmeh,3 Muzammil Khan2 1Department of Medicine, Lourdes Hospital, Binghamton, NY, USA, 2Department of Medicine, NYC Health + Hospital/Harlem, Columbia University, NY, USA, 3Department of Medicine, Lincoln Medical and Mental Health Centre, Bronx NY, USA Background: Epstein–Barr virus (EBV has been found to cause infectious mononucleosis multiple times, but has been associated rarely with EBV encephalitis. Also, whenever it is diagnosed, it is always treated symptomatically.Case report: A case of confirmed EBV encephalitis is presented, which was treated with antiviral therapy resulting in complete clearance of the virus in cerebrospinal fluid and minimal neurologic symptoms after hospital discharge.Conclusion: The Infectious Diseases Society of America guidelines state that intravenous acyclovir is not recommended for EBV-related encephalitis. But we reviewed the literature and found similar cases, and we believe that antiviral therapy should be recommended for EBV encephalitis because it is a potentially fatal disease and if left untreated, can lead to raised intracranial pressure, craniotomy and even death. Keywords: Epstein–Barr virus, intravenous, human immune deficiency virus, HIV

  16. Use of different marker pre-selection methods based on single SNP regression in the estimation of Genomic-EBVs

    Directory of Open Access Journals (Sweden)

    Corrado Dimauro

    2010-01-01

    Full Text Available Two methods of SNPs pre-selection based on single marker regression for the estimation of genomic breeding values (G-EBVs were compared using simulated data provided by the XII QTL-MAS workshop: i Bonferroni correction of the significance threshold and ii Permutation test to obtain the reference distribution of the null hypothesis and identify significant markers at P<0.01 and P<0.001 significance thresholds. From the set of markers significant at P<0.001, random subsets of 50% and 25% markers were extracted, to evaluate the effect of further reducing the number of significant SNPs on G-EBV predictions. The Bonferroni correction method allowed the identification of 595 significant SNPs that gave the best G-EBV accuracies in prediction generations (82.80%. The permutation methods gave slightly lower G-EBV accuracies even if a larger number of SNPs resulted significant (2,053 and 1,352 for 0.01 and 0.001 significance thresholds, respectively. Interestingly, halving or dividing by four the number of SNPs significant at P<0.001 resulted in an only slightly decrease of G-EBV accuracies. The genetic structure of the simulated population with few QTL carrying large effects, might have favoured the Bonferroni method.

  17. Mitochondrial DNA single nucleotide polymorphism associated with weight estimated breeding values in Nelore cattle (Bos indicus

    Directory of Open Access Journals (Sweden)

    Fernando Henrique Biase

    2007-01-01

    Full Text Available We sampled 119 Nelore cattle (Bos indicus, 69 harboring B. indicus mtDNA plus 50 carrying Bos taurus mtDNA, to estimate the frequencies of putative mtDNA single nucleotide polymorphisms (SNPs and investigate their association with Nelore weight and scrotal circumference estimated breeding values (EBVs. The PCR restriction fragment length polymorphism (PCR-RFLP method was used to detect polymorphisms in the mitochondrial asparagine, cysteine, glycine, leucine and proline transporter RNA (tRNA genes (tRNAasn, tRNAcys, tRNAgly, tRNAleu and tRNApro. The 50 cattle carrying B. taurus mtDNA were monomorphic for all the tRNA gene SNPs analyzed, suggesting that they are specific to mtDNA from B. indicus cattle. No tRNAcys or tRNAgly polymorphisms were detected in any of the cattle but we did detect polymorphic SNPs in the tRNAasn, tRNAleu and tRNApro genes in the cattle harboring B. indicus mtDNA, with the same allele observed in the B. taurus sequence being present in the following percentage of cattle harboring B. indicus mtDNA: 72.46% for tRNAasn, 95.23% for tRNAleu and 90.62% for tRNApro. Analyses of variance using the tRNAasn SNP as the independent variable and EBVs as the dependent variable showed that the G -> T SNP was significantly associated (p < 0.05 with maternal EBVs for weight at 120 and 210 days (p < 0.05 and animal's EBVs for weight at 210, 365 and 455 days. There was no association of the tRNAasn SNP with the scrotal circumference EBVs. These results confirm that mtDNA can affect weight and that mtDNA polymorphisms can be a source of genetic variation for quantitative traits.

  18. Comparison of DNA extraction methods from small samples of newborn screening cards suitable for retrospective perinatal viral research.

    Science.gov (United States)

    McMichael, Gai L; Highet, Amanda R; Gibson, Catherine S; Goldwater, Paul N; O'Callaghan, Michael E; Alvino, Emily R; MacLennan, Alastair H

    2011-04-01

    Reliable detection of viral DNA in stored newborn screening cards (NSC) would give important insight into possible silent infection during pregnancy and around birth. We sought a DNA extraction method with sufficient sensitivity to detect low copy numbers of viral DNA from small punch samples of NSC. Blank NSC were spotted with seronegative EDTA-blood and seropositive EBV EDTA-blood. DNA was extracted with commercial and noncommercial DNA extraction methods and quantified on a spectrofluorometer using a PicoGreen dsDNA quantification kit. Serial dilutions of purified viral DNA controls determined the sensitivity of the amplification protocol, and seropositive EBV EDTA-blood amplified by nested PCR (nPCR) validated the DNA extraction methods. There were considerable differences between the commercial and noncommercial DNA extraction methods (P=0.014; P=0.016). Commercial kits compared favorably, but the QIamp DNA micro kit with an added forensic filter step was marginally more sensitive. The mean DNA yield from this method was 3 ng/μl. The limit of detection was 10 viral genome copies in a 50-μl reaction. EBV nPCR detection in neat and 1:10 diluted DNA extracts could be replicated reliably. We conclude that the QIamp Micro DNA extraction method with the added forensic spin-filter step was suitable for retrospective DNA viral assays from NSC.

  19. Nonhazardous Urine Pretreatment Method

    Science.gov (United States)

    Akse, James R.; Holtsnider, John T.

    2012-01-01

    A method combines solid phase acidification with two non-toxic biocides to prevent ammonia volatilization and microbial proliferation. The safe, non-oxidizing biocide combination consists of a quaternary amine and a food preservative. This combination has exhibited excellent stabilization of both acidified and unacidified urine. During pretreatment tests, composite urine collected from donors was challenged with a microorganism known to proliferate in urine, and then was processed using the nonhazardous urine pre-treatment method. The challenge microorganisms included Escherichia coli, a common gram-negative bacteria; Enterococcus faecalis, a ureolytic gram-positive bacteria; Candida albicans, a yeast commonly found in urine; and Aspergillus niger, a problematic mold that resists urine pre-treatment. Urine processed in this manner remained microbially stable for over 57 days. Such effective urine stabilization was achieved using non-toxic, non-oxidizing biocides at higher pH (3.6 to 5.8) than previous methods in use or projected for use aboard the International Space Station (ISS). ISS urine pretreatment methods employ strong oxidants including ozone and hexavalent chromium (Cr(VI)), a carcinogenic material, under very acidic conditions (pH = 1.8 to 2.4). The method described here offers a much more benign chemical environment than previous pretreatment methods, and will lower equivalent system mass (ESM) by reducing containment volume and mass, system complexity, and crew time needed to handle pre-treatment chemicals. The biocides, being non-oxidizing, minimize the potential for chemical reactions with urine constituents to produce volatile, airborne contaminants such as cyanogen chloride. Additionally, the biocides are active under significantly less acidic conditions than those used in the current system, thereby reducing the degree of required acidification. A simple flow-through solid phase acidification (SPA) bed is employed to overcome the natural buffering

  20. Frequency of EBV associated classical Hodgkin lymphoma decreases over a 54-year period in a Brazilian population.

    Science.gov (United States)

    Campos, Antonio Hugo Jose Froes Marques; Moreira, Adriana; Ribeiro, Karina Braga; Paes, Roberto Pinto; Zerbini, Maria Claudia; Aldred, Vera; de Souza, Carmino Antonio; Neto, Cristovam Scapulatempo; Soares, Fernando Augusto; Vassallo, Jose

    2018-01-30

    The epidemiology of classical Hodgkin lymphoma varies significantly in populations with different socioeconomic conditions. Among other changes, improvement in such conditions leads to a reduction in the association with EBV infection and predominance of the nodular sclerosis subtype. This study provides an overview of the epidemiology of 817 cases of classical Hodgkin lymphoma diagnosed in five reference hospitals of the State of Sao Paulo, Brazil, over 54 years (1954-2008). The cases were distributed in 3 periods (1954-1979; 1980-1999; and 2000-2008). EBV-positive cases decreased from 87% to 46%. In children and adolescents (45 years), the decrease in EBV infection was not significant. Nodular Sclerosis was the most common subtype in all periods. These results support the hypothesis that, in the Brazilian State of Sao Paulo, classical Hodgkin lymphoma has changed and now shows characteristics consistent with Pattern III observed in populations that experienced a similar socioeconomic transition.

  1. The Epstein-Barr virus DNA load in the peripheral blood of transplant recipients does not accurately reflect the burden of infected cells.

    Science.gov (United States)

    Fink, Susanne; Tsai, Ming-Han; Schnitzler, Paul; Zeier, Martin; Dreger, Peter; Wuchter, Patrick; Bulut, Olcay C; Behrends, Uta; Delecluse, Henri-Jacques

    2017-01-01

    Transplant recipients frequently exhibit an increased Epstein-Barr virus (EBV) load in the peripheral blood. Here, we quantitated the EBV-infected cells in the peripheral blood of these patients and defined the mode of viral infection, latent or lytic. These data indicated that there is no strong correlation between the number of infected cells and the EBV load (EBVL). This can be explained by a highly variable number of EBV copies per infected cell and by lytic replication in some cells. The plasma of these patients did not contain any free infectious viruses, but contained nevertheless EBV DNA, sometimes in large amounts, that probably originates from cell debris and contributed to the total EBVL. Some of the investigated samples carried a highly variable number of infected cells in active latency, characterized by an expression of the Epstein-Barr nuclear antigens (EBNA2) protein. However, a third of the samples expressed neither EBNA2 nor lytic proteins. Patients with an increased EBVL represent a heterogeneous group of patients whose infection cannot be characterized by this method alone. Precise characterization of the origin of an increased EBVL, in particular, in terms of the number of EBV-infected cells, requires additional investigations including the number of EBV-encoded small RNA-positive cells. © 2016 Steunstichting ESOT.

  2. Biomass shock pretreatment

    Science.gov (United States)

    Holtzapple, Mark T.; Madison, Maxine Jones; Ramirez, Rocio Sierra; Deimund, Mark A.; Falls, Matthew; Dunkelman, John J.

    2014-07-01

    Methods and apparatus for treating biomass that may include introducing a biomass to a chamber; exposing the biomass in the chamber to a shock event to produce a shocked biomass; and transferring the shocked biomass from the chamber. In some aspects, the method may include pretreating the biomass with a chemical before introducing the biomass to the chamber and/or after transferring shocked biomass from the chamber.

  3. A spectrum of basaloid morphology in a subset of EBV-associated "lymphoepithelial carcinomas" of major salivary glands

    DEFF Research Database (Denmark)

    Friborg, Jeppe Tang; Therkildsen, Marianne Hamilton; Homoe, Preben

    2012-01-01

    salivary gland carcinomas. Amongst primary LEC of major salivary gland, most cases reported in the literature have represented typical nasopharynx-like tumors. Variants of Epstein-Barr Virus (EBV) associated LEC have not been described previously, to the best of our knowledge. In this report, we describe 4...... EBV-associated major salivary gland LECs with prominent basaloid morphology, which represent 22 % of a cohort of 18 salivary LECs from an Inuit population in Greenland. The features described in these cases raise a differential diagnosis of other basaloid tumors, particularly in light of the salivary...

  4. Epigenetic control of viral life-cycle by a DNA-methylation dependent transcription factor.

    Directory of Open Access Journals (Sweden)

    Kirsty Flower

    Full Text Available Epstein-Barr virus (EBV encoded transcription factor Zta (BZLF1, ZEBRA, EB1 is the prototype of a class of transcription factor (including C/EBPalpha that interact with CpG-containing DNA response elements in a methylation-dependent manner. The EBV genome undergoes a biphasic methylation cycle; it is extensively methylated during viral latency but is reset to an unmethylated state following viral lytic replication. Zta is expressed transiently following infection and again during the switch between latency and lytic replication. The requirement for CpG-methylation at critical Zta response elements (ZREs has been proposed to regulate EBV replication, specifically it could aid the activation of viral lytic gene expression from silenced promoters on the methylated genome during latency in addition to preventing full lytic reactivation from the non-methylated EBV genome immediately following infection. We developed a computational approach to predict the location of ZREs which we experimentally assessed using in vitro and in vivo DNA association assays. A remarkably different binding motif is apparent for the CpG and non-CpG ZREs. Computational prediction of the location of these binding motifs in EBV revealed that the majority of lytic cycle genes have at least one and many have multiple copies of methylation-dependent CpG ZREs within their promoters. This suggests that the abundance of Zta protein coupled with the methylation status of the EBV genome act together to co-ordinate the expression of lytic cycle genes at the majority of EBV promoters.

  5. Epstein-Barr virus antibodies in serum and DNA load in saliva are not associated with radiological or clinical disease activity in patients with early multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    René M Gieß

    Full Text Available To investigate the association of Epstein-Barr virus (EBV nuclear antigen-1 (EBNA-1 and viral capsid antigen (VCA immunoglobulin (IgG antibodies in serum as well as EBV DNA load in saliva with radiological and clinical disease activity in patients with clinically isolated syndrome (CIS and early relapsing-remitting MS (RRMS.EBNA-1 and VCA immunoglobulin (IgG antibodies were determined in serum of 100 patients with CIS/early RRMS and 60 healthy controls. EBV DNA load was measured in saliva of 48 patients and 50 controls. Patients underwent clinical assessment with the Expanded Disability Status Scale (EDSS and 3 Tesla magnetic resonance imaging at baseline and after a median of 20 months of follow-up (n = 63 for MRI, n = 71 for EDSS. The association of EBV parameters with occurrence of a second relapse, indicating conversion to clinically definite MS (CDMS, was evaluated over a median of 35 months of follow-up after the first clinical event (n = 89.EBNA-1 IgG antibody frequency (p = 0.00005 and EBNA-1 and VCA IgG antibody levels (p<0.0001 for both were higher in patients than in controls. EBV DNA load in saliva did not differ between groups. Neither EBV antibody levels nor DNA load in saliva were associated with baseline or follow-up number or volume of T2-weighted (T2w or contrast enhancing lesions, number of Barkhof criteria or the EDSS, or with the number of new T2w lesions, T2w lesion volume change or EDSS change on follow-up. Likewise, levels of EBV IgG antibodies in serum and DNA load in saliva were not associated with conversion to CDMS.While these findings confirm the association of EBV infection with early MS, neither EBNA-1 nor VCA IgG antibodies in serum nor EBV DNA load in saliva were associated with radiological or clinical disease activity in patients with CIS/early RRMS. These data are compatible with the concept that EBV may be a trigger for MS acting very early during the development of the disease.

  6. Reconstitution of EBV latent but not lytic antigen-specific CD4(+) and CD8(+) T cells after HIV treatment with highly active antiretroviral therapy

    NARCIS (Netherlands)

    Piriou, Erwan; Jansen, Christine A.; van Dort, Karel; de Cuyper, Iris; Nanlohy, Nening M.; Lange, Joep M. A.; van Oers, Marinus H. J.; Miedema, Frank; van Baarle, Debbie

    2005-01-01

    The incidence of (EBV-rlelated) malignancies in HIV-infected subjects has declined since the introduction of highly active antiretroviral therapy (HAART). To investigate the effect of HAART on EBV infection, we performed a longitudinal analysis of the T cell response to both a latent and a lytic Ag

  7. Aberrant Expression of ID2 protein and its correlation with EBV-LMP1 and P16(INK4A) in Classical Hodgkin Lymphoma in China

    International Nuclear Information System (INIS)

    Zhao, Po; Lu, Yali; Liu, Lin; Zhong, Mei

    2008-01-01

    The relationships between the expression of ID2, EBV-LMP1 and P16(INK4A) in Chinese classical Hodgkin lymphoma are unknown and need exploring. Samples of classical Hodgkin lymphoma from 60 Chinese patients were analyzed for the expression of ID2, EBV-LMP1 and p16(INK4A) proteins by immunohistochemistry. ID2 protein was expressed in 83.3% of this group of classical Hodgkin lymphoma, staining strongly in both cytoplasm and nucleus of the Hodgkin and Reed-Sternberg (HRS) cells. EBV-LMP1 and P16(INK4A) were overexpressed in 85.0% and 71.7% of Hodgkin lymphoma, respectively. EBV-LMP1 was noted in the cytoplasm, membrane and nucleus of HRS cells; P16(INK4A) was in the nucleus and cytoplasm. Microscopically, ID2, EBV-LMP1 and P16(INK4A) staining distinguished the HRS cells from the complex background of lymphocytes. ID2 was positively correlated with EBV-LMP1(P < 0.01), but P16(INK4A) was inversely related to EBV-LMP1 (P < 0.05). It is suggested that ID2, EBV-LMP1 and P16(INK4A) could play an important role in the evolution of classical Hodgkin lymphoma, and be considered as potential adjunct markers to identify HRS cells in diagnosis

  8. Discordance of epstein-barr virus (ebv) specific humoral and cellular immunity in patients with malignant lymphomas : Elevated antibody titers and lowered invitro lymphocyte-reactivity

    NARCIS (Netherlands)

    ten Napel, C. H. H.; The, T. Hauw; van Egten-Bijker, J; de Gast, G. C.; Halie, M. R.; Langenhuysen, M. M. A. C.

    1978-01-01

    The relationship between specific viral cellular and humoral immunity to the Epstein–Barr Virus (EBV) was investigated in thirty-one untreated patients with malignant lymphoma (ML) and sex- and age-matched controls. In vitro reactivity of peripheral blood lymphocytes to heatinactivated purified EBV,

  9. The EBV immunoevasins vIL-10 and BNLF2a protect newly infected B cells from immune recognition and elimination.

    Directory of Open Access Journals (Sweden)

    Simon Jochum

    Full Text Available Lifelong persistence of Epstein-Barr virus (EBV in infected hosts is mainly owed to the virus' pronounced abilities to evade immune responses of its human host. Active immune evasion mechanisms reduce the immunogenicity of infected cells and are known to be of major importance during lytic infection. The EBV genes BCRF1 and BNLF2a encode the viral homologue of IL-10 (vIL-10 and an inhibitor of the transporter associated with antigen processing (TAP, respectively. Both are known immunoevasins in EBV's lytic phase. Here we describe that BCRF1 and BNLF2a are functionally expressed instantly upon infection of primary B cells. Using EBV mutants deficient in BCRF1 and BNLF2a, we show that both factors contribute to evading EBV-specific immune responses during the earliest phase of infection. vIL-10 impairs NK cell mediated killing of infected B cells, interferes with CD4+ T-cell activity, and modulates cytokine responses, while BNLF2a reduces antigen presentation and recognition of newly infected cells by EBV-specific CD8+ T cells. Together, both factors significantly diminish the immunogenicity of EBV-infected cells during the initial, pre-latent phase of infection and may improve the establishment of a latent EBV infection in vivo.

  10. Analysis of the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP-1) gene and promoter in Hodgkin's disease isolates

    DEFF Research Database (Denmark)

    Sandvej, K; Andresen, B S; Zhou, X G

    2000-01-01

    AIMS: To study the distribution of Epstein-Barr virus (EBV) variants containing mutations in the latent membrane protein 1 (LMP-1) oncogene and promoter in EBV associated Hodgkin's disease and infectious mononucleosis compared with previous findings in asymptomatic EBV carriers. METHODS: Sequence...... analysis of the EBV LMP-1 promoter and gene in isolates from Danish patients with Hodgkin's disease (n = 61) and infectious mononucleosis (n = 10). RESULTS: Viruses (previously designated group D) that contain two mutations in the activating transcription factor/cAMP response element (ATF/CRE) in the LMP-1...... promoter, which are known to decrease promoter activity greatly, were significantly less frequent in Hodgkin's disease than in both infectious mononucleosis (p = 0.0081) and asymptomatic EBV carriers (p = 0.0084). In some cases, the LMP-1 gene contained mutations in a recently identified cytotoxic T cell...

  11. Mechanism of melphalan crosslink enhancement by misonidazole pretreatment

    International Nuclear Information System (INIS)

    Taylor, Y.C.; Sawyer, J.M.; Hsu, B.; Brown, J.M.

    1984-01-01

    Sensitization of Chinese hamster ovary cells to melphalan (L-PAM) toxicity by prior treatment with misonidazole is associated with increased levels of DNA crosslinks believed to be the critical lesion for bifunctional alkylating agent toxicity. Enhanced L-PAM crosslinking of DNA could occur by a variety of mechanisms in MISO-pretreated cells including: (1) increased transport or binding of L-PAM, (2) decreased repair of L-PAM monoadducts which would allow more time for their conversion to crosslinks, (3) decreased crosslink repair (unhooking of one arm), or (4) chemical modification of the DNA structure, presumably by bound MISO derivatives, such that crosslink formation is facilitated. Previous studies have eliminated mechanisms (1) and (3). Mechanism (4) was investigated by following MISO-pretreatments of whole cells with L-PAM treatments of the isolated DNA from these cells. Treatment of bare DNA with L-PAM modeled very well the crosslinking behavior in whole cells although it was somewhat more efficient. In the presence of double stranded DNA and absence of repair systems during and after the L-PAM exposure, it was determined that MISO-pretreatments did not increase the crosslinking efficiency of L-PAM

  12. Plasma Epstein-Barr virus and Hepatitis B virus in non-Hodgkin lymphomas: Two lymphotropic, potentially oncogenic, latently occurring DNA viruses.

    Science.gov (United States)

    Sinha, Mahua; Rao, Clementina Rama; Premalata, C S; Shafiulla, Mohammed; Lakshmaiah, K C; Jacob, Linu Abraham; Babu, Govind K; Viveka, B K; Appaji, L; Subramanyam, Jayshree R

    2016-01-01

    There is a need to study potential infective etiologies in lymphomas. Lymphocyte-transforming viruses can directly infect lymphocytes, disrupt normal cell functions, and promote cell division. Epstein-Barr virus (EBV) is known to be associated with several lymphomas, especially Hodgkin lymphomas (HLs). And recently, the lymphocyte-transforming role of hepatitis B virus (HBV) has been emphasized. The aim of this study was to elucidate the association of two potentially oncogenic, widely prevalent latent DNA viruses, EBV and HBV, in non-HL (NHL). In this prospective study, we estimated plasma EBV and HBV DNA in NHL patients. Peripheral blood was obtained from newly diagnosed, treatment na ïve, histologically confirmed NHL patients. Plasma EBV DNA was quantified by real-time polymerase chain reaction (PCR) targeting Epstein-Barr Nucleic acid 1 while the plasma HBV DNA was detected using nested PCR targeting HBX gene. In a small subset of patients, follow-up plasma samples post-anticancer chemotherapy were available and retested for viral DNA. Of the 110 NHL patients, ~79% were B-cell NHL and ~21% were T-cell NHL. Plasma EBV-DNA was detected in 10% NHLs with a higher EBV association in Burkitt lymphoma (33.3%) than other subtypes. Pretherapy HBV DNA was detected in 21% NHLs; most of them being diffuse large B-cell lymphoma (DLBCL). Moreover, 42% of DLBCL patients had HBV DNA in plasma. Since all patients were HBV surface antigen seronegative at diagnosis, baseline plasma HBV-DNAemia before chemotherapy was indicative of occult hepatitis B infection. Our findings indicate a significant association of HBV with newly diagnosed DLBCL.

  13. (HHV6) et d'Epstein-Barr virus (EBV) par PCR en temps réel chez ...

    African Journals Online (AJOL)

    Diagnostic moléculaire du Cytomégalovirus (CMV), de l'herpès virus humain de type 6 (HHV6) et d'Epstein-Barr virus (EBV) par PCR en temps réel chez les femmes enceintes VIH séropositives et séronégatives à Ouagadougou, Burkina Faso.

  14. Inhibition of EBV-mediated membrane fusion by anti-gHgL antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Sathiyamoorthy, Karthik; Jiang, Jiansen; Möhl, Britta S.; Chen, Jia; Zhou, Z. Hong; Longnecker, Richard; Jardetzky, Theodore S. (UCLA); (Stanford-MED); (NWU)

    2017-09-22

    Herpesvirus entry into cells requires the coordinated action of multiple virus envelope glycoproteins, including gH, gL, and gB. For EBV, the gp42 protein assembles into complexes with gHgL heterodimers and binds HLA class II to activate gB-mediated membrane fusion with B cells. EBV tropism is dictated by gp42 levels in the virion, as it inhibits entry into epithelial cells while promoting entry into B cells. The gHgL and gB proteins are targets of neutralizing antibodies and potential candidates for subunit vaccine development, but our understanding of their neutralizing epitopes and the mechanisms of inhibition remain relatively unexplored. Here we studied the structures and mechanisms of two anti-gHgL antibodies, CL40 and CL59, that block membrane fusion with both B cells and epithelial cells. We determined the structures of the CL40 and CL59 complexes with gHgL using X-ray crystallography and EM to identify their epitope locations. CL59 binds to the C-terminal domain IV of gH, while CL40 binds to a site occupied by the gp42 receptor binding domain. CL40 binding to gHgL/gp42 complexes is not blocked by gp42 and does not interfere with gp42 binding to HLA class II, indicating that its ability to block membrane fusion with B cells represents a defect in gB activation. These data indicate that anti-gHgL neutralizing antibodies can block gHgL-mediated activation of gB through different surface epitopes and mechanisms.

  15. DNA Oncogenic Virus-Induced Oxidative Stress, Genomic Damage, and Aberrant Epigenetic Alterations

    Directory of Open Access Journals (Sweden)

    Mankgopo Magdeline Kgatle

    2017-01-01

    Full Text Available Approximately 20% of human cancers is attributable to DNA oncogenic viruses such as human papillomavirus (HPV, hepatitis B virus (HBV, and Epstein-Barr virus (EBV. Unrepaired DNA damage is the most common and overlapping feature of these DNA oncogenic viruses and a source of genomic instability and tumour development. Sustained DNA damage results from unceasing production of reactive oxygen species and activation of inflammasome cascades that trigger genomic changes and increased propensity of epigenetic alterations. Accumulation of epigenetic alterations may interfere with genome-wide cellular signalling machineries and promote malignant transformation leading to cancer development. Untangling and understanding the underlying mechanisms that promote these detrimental effects remain the major objectives for ongoing research and hope for effective virus-induced cancer therapy. Here, we review current literature with an emphasis on how DNA damage influences HPV, HVB, and EBV replication and epigenetic alterations that are associated with carcinogenesis.

  16. Effect of cadmium pretreatment on liver regeneration after partial hepatectomy in rats

    International Nuclear Information System (INIS)

    Margeli, A.; Theocharis, S.; Skaltsas, S.; Skopelitou, A.; Kittas, C.; Mykoniatis, M.; Varonos, D.

    1994-01-01

    In this study we examined the effect of cadmium pretreatment, administered 24 h before partial hepatectomy, on the liver regenerative process in rats, at different time intervals. The rate of 3 H thymidine incorporation into hepatic DNA and the activity of the enzyme thymidine kinase were used as indices of liver proliferative capacity. Thymidine kinase, the rate-determining enzyme of DNA biosynthesis, was suppressed during the first hours following partial hepatectomy in the liver of cadmium pretreated animals. DNA biosynthesis was also strongly decreased in cadmium pretreated animals, by delaying the first peak of liver regeneration, compared with the partially hepatectomized ones. Biochemical parameters, mitotic index and proliferating cell nuclear antigen staining were also coestimated. The above data suggest that cadmium pretreatment suppressed the hepatic regenerative process, probably due to the inhibition of thymidine kinase. (orig./MG)

  17. Effect of cadmium pretreatment on liver regeneration after partial hepatectomy in rats

    Energy Technology Data Exchange (ETDEWEB)

    Margeli, A. (Dept. of Experimental Pharmacology, Medical School, Univ. Athens (Greece)); Theocharis, S. (Dept. of Experimental Pharmacology, Medical School, Univ. Athens (Greece)); Skaltsas, S. (Dept. of Experimental Pharmacology, Medical School, Univ. Athens (Greece)); Skopelitou, A. (Dept. of Pathology, Medical School, Univ. Athens (Greece)); Kittas, C. (Dept. of Pathology, Medical School, Univ. Athens (Greece)); Mykoniatis, M. (Dept. of Experimental Pharmacology, Medical School, Univ. Athens (Greece)); Varonos, D. (Dept. of Experimental Pharmacology, Medical School, Univ. Athens (Greece))

    1994-02-01

    In this study we examined the effect of cadmium pretreatment, administered 24 h before partial hepatectomy, on the liver regenerative process in rats, at different time intervals. The rate of [sup 3]H thymidine incorporation into hepatic DNA and the activity of the enzyme thymidine kinase were used as indices of liver proliferative capacity. Thymidine kinase, the rate-determining enzyme of DNA biosynthesis, was suppressed during the first hours following partial hepatectomy in the liver of cadmium pretreated animals. DNA biosynthesis was also strongly decreased in cadmium pretreated animals, by delaying the first peak of liver regeneration, compared with the partially hepatectomized ones. Biochemical parameters, mitotic index and proliferating cell nuclear antigen staining were also coestimated. The above data suggest that cadmium pretreatment suppressed the hepatic regenerative process, probably due to the inhibition of thymidine kinase. (orig./MG)

  18. Measurement of Epstein-Barr virus DNA loads in whole blood and plasma by TaqMan PCR and in peripheral blood lymphocytes by competitive PCR.

    Science.gov (United States)

    Wadowsky, Robert M; Laus, Stella; Green, Michael; Webber, Steven A; Rowe, David

    2003-11-01

    Epstein-Barr virus (EBV) DNA load values were measured in samples of whole blood (n = 60) and plasma (n = 59) by TaqMan PCR and in samples of peripheral blood lymphocytes (PBLs) (n = 60) by competitive PCR (cPCR). The samples were obtained from 44 transplant recipients. The whole-blood and PBL loads correlated highly (r(2) > 0.900), whereas the plasma and PBL loads correlated poorly (r(2) = 0.512). Testing of whole blood by TaqMan PCR is an acceptable alternative to testing of PBLs by cPCR for quantifying EBV DNA load.

  19. Changes in Tonsil B Cell Phenotypes and EBV Receptor Expression in Children Under 5-Years-Old.

    Science.gov (United States)

    Wohlford, Eric M; Baresel, Paul C; Wilmore, Joel R; Mortelliti, Anthony J; Coleman, Carrie B; Rochford, Rosemary

    2018-03-01

    Palatine tonsils are principally B cell organs that are the initial line of defense against many oral pathogens, as well as the site of infection for others. While the size of palatine tonsils changes greatly in the first five years of life, the cellular changes during this period are not well studied. Epstein Barr virus (EBV) is a common orally transmitted virus that infects tonsillar B cells. Naïve B cells are thought to be the target of primary infection with EBV in vivo, suggesting that they are targeted by the virus. EBV enters B cells through CD21, but studies of older children and adults have not shown differences in surface CD21 between naïve B cells and other tonsil B cell populations. In this study, we used an 11-color flow cytometry panel to detail the changes in B cell subpopulations in human tonsils over the first five years of life from 33 healthy US children. We provide reference ranges for tonsil B cell subpopulations over this age range. We show that the frequency of naïve tonsil B cells decreases over the early years of life, and that naïve B cells expressed higher surface levels of CD21 relative to other tonsil B cell populations. We show that young children have a higher frequency of naïve tonsil B cells, and importantly that these cells express increased surface EBV receptor, suggesting that young children have a larger pool of cells that can be infected by the virus. © 2017 International Clinical Cytometry Society. © 2017 International Clinical Cytometry Society.

  20. Improving nuclear envelope dynamics by EBV BFRF1 facilitates intranuclear component clearance through autophagy.

    Science.gov (United States)

    Liu, Guan-Ting; Kung, Hsiu-Ni; Chen, Chung-Kuan; Huang, Cheng; Wang, Yung-Li; Yu, Cheng-Pu; Lee, Chung-Pei

    2018-02-26

    Although a vesicular nucleocytoplasmic transport system is believed to exist in eukaryotic cells, the features of this pathway are mostly unknown. Here, we report that the BFRF1 protein of the Epstein-Barr virus improves vesicular transport of nuclear envelope (NE) to facilitate the translocation and clearance of nuclear components. BFRF1 expression induces vesicles that selectively transport nuclear components to the cytoplasm. With the use of aggregation-prone proteins as tools, we found that aggregated nuclear proteins are dispersed when these BFRF1-induced vesicles are formed. BFRF1-containing vesicles engulf the NE-associated aggregates, exit through from the NE, and putatively fuse with autophagic vacuoles. Chemical treatment and genetic ablation of autophagy-related factors indicate that autophagosome formation and autophagy-linked FYVE protein-mediated autophagic proteolysis are involved in this selective clearance of nuclear proteins. Remarkably, vesicular transport, elicited by BFRF1, also attenuated nuclear aggregates accumulated in neuroblastoma cells. Accordingly, induction of NE-derived vesicles by BFRF1 facilitates nuclear protein translocation and clearance, suggesting that autophagy-coupled transport of nucleus-derived vesicles can be elicited for nuclear component catabolism in mammalian cells.-Liu, G.-T., Kung, H.-N., Chen, C.-K., Huang, C., Wang, Y.-L., Yu, C.-P., Lee, C.-P. Improving nuclear envelope dynamics by EBV BFRF1 facilitates intranuclear component clearance through autophagy.

  1. Influence of Maximum Inbreeding Avoidance under BLUP EBV Selection on Pinzgau Population Diversity

    Directory of Open Access Journals (Sweden)

    Radovan Kasarda

    2011-05-01

    Full Text Available Evaluated was effect of mating (random vs. maximum avoidance of inbreeding under BLUP EBV selection strategy. Existing population structure was under Monte Carlo stochastic simulation analyzed from the point to minimize increase of inbreeding. Maximum avoidance of inbreeding under BLUP selection resulted into comparable increase of inbreeding then random mating in average of 10 generation development. After 10 generations of simulation of mating strategy was observed ΔF= 6,51 % (2 sires, 5,20 % (3 sires, 3,22 % (4 sires resp. 2,94 % (5 sires. With increased number of sires selected, decrease of inbreeding was observed. With use of 4, resp. 5 sires increase of inbreeding was comparable to random mating with phenotypic selection. For saving of genetic diversity and prevention of population loss is important to minimize increase of inbreeding in small populations. Classical approach was based on balancing ratio of sires and dams in mating program. Contrariwise in the most of commercial populations small number of sires was used with high mating ratio.

  2. A workflow for in silico design of hIL-10 and ebvIL-10 inhibitors using well-known miniprotein scaffolds.

    Science.gov (United States)

    Dueñas, Salvador; Aguila, Sergio A; Pimienta, Genaro

    2017-04-01

    The over-expression of immune-suppressors such as IL-10 is a crucial landmark in both tumor progression, and latent viral and parasite infection. IL-10 is a multifunctional protein. Besides its immune-cell suppressive function, it also promotes B-cell tumorigenesis of lymphomas and melanoma. Human pathogens like unicellular parasites and viruses that remain latent inside B cells promote the over-expression of hIL-10 upon infection, which inhibits cell-mediated immune surveillance, and at the same time mediates B cell proliferation. The B-cell specific oncogenic latent virus Epstein-Barr virus (EBV) encodes a viral homologue of hIL-10 (ebvIL-10), expressed during lytic viral proliferation. Once expressed, ebvIL-10 inhibits cell-mediated immune surveillance, assuring EBV re-infection. During long-term latency, EBV-infected B cells over-express hIL-10 to assure B-cell proliferation, occasionally inducing EBV-mediated lymphomas. The amino acid sequences of hIL-10 and ebvIL-10 are more than 80% identical and thus have a very similar tridimensional structure. Based on their published crystallographic structures bound to their human receptor IL10R1, we report a structure-based design of hIL-10 and ebvIL-10 inhibitors based on 3 loops from IL10R1 that establish specific hydrogen bonds with the two IL10s. We have grafted these loops onto a permissible loop in three well-known miniprotein scaffolds-the Conus snail toxin MVIIA, the plant-derived trypsin inhibitor EETI, and the human appetite modulator AgRP. Our computational workflow described in detail below was invigorated by the negative and positive controls implemented, and therefore paves the way for future in vitro and in vivo validation assays of the IL-10 inhibitors engineered.

  3. Measurement of CD8+ and CD4+ T Cell Frequencies Specific for EBV LMP1 and LMP2a Using mRNA-Transfected DCs.

    Directory of Open Access Journals (Sweden)

    Dae-Hee Sohn

    Full Text Available An EBV-specific cellular immune response is associated with the control of EBV-associated malignancies and lymphoproliferative diseases, some of which have been successfully treated by adoptive T cell therapy. Therefore, many methods have been used to measure EBV-specific cellular immune responses. Previous studies have mainly used autologous EBV-transformed B-lymphoblastoid cell lines (B-LCLs, recombinant viral vectors transfected or peptide pulsed dendritic cells (DCs as stimulators of CD8(+ and CD4(+ T lymphocytes. In the present study, we used an interferon-γ (IFN-γ enzyme-linked immunospot (ELISPOT assay by using isolated CD8(+ and CD4(+ T cells stimulated with mRNA-transfected DCs. The frequency of latent membrane protein 1 (LMP1-specific IFN-γ producing CD4(+ T cells was significantly higher than that of LMP2a. The frequency of IFN-γ producing CD4(+ T cells was significantly correlated with that of CD8(+ T cells in LMP1-specific immune responses (r = 0.7187, Pc < 0.0001. To determine whether there were changes in LMP1- or LMP2a-specific immune responses, subsequent peripheral blood mononuclear cells (PBMCs samples were analyzed. Significant changes were observed in 5 of the 10 donors examined, and CD4(+ T cell responses showed more significant changes than CD8(+ T cell responses. CD8(+ and CD4(+ T cells from EBV-seropositive donors secreted only the Th1 cytokines IFN-γ, TNF-α, and IL-2, while Th2 (IL-4 and Th17 (IL-17a cytokines were not detected. CD4(+ T cells secreted significantly higher cytokine levels than did CD8(+ T cells. Analysis of EBV-specific T cell responses using autologous DCs transfected with mRNA might provide a comprehensive tool for monitoring EBV infection and new insights into the pathogenesis of EBV-associated diseases.

  4. Human-derived IgG level as an indicator for EBV-associated lymphoma model in Hu-PBL/SCID chimeras.

    Science.gov (United States)

    Tang, Yunlian; He, Rongfang; Zhang, Yang; Liu, Fang; Cheng, Ailan; Wu, Yimou; Gan, Runliang

    2011-05-09

    Epstein-Barr virus (EBV) has a close association with various types of human lymphomas. Animal models are essential to elucidate the pathogenesis of human EBV-associated lymphomas. The aim of the present study is to evaluate the association between human IgG concentration and EBV-associated lymphoma development in huPBL/SCID mice. Human peripheral blood lymphocytes (hu-PBL) from EBV-seropositive donors were inoculated intraperitoneally into SCID mouse. Immunohistochemical staining was used to examine differentiated antigens of tumor cells. EBV infection of the induced tumors was detected by in situ hybridization. IgG concentrations in the serums of 12 SCID mice were measured by unidirectional immunodiffusion assay. 21 out of 29 mice developed tumors in their body. Immunohistochemical staining showed that all induced tumors were LCA (leukocyte common antigen) positive, B-cell markers (CD20, CD79a) positive, and T-cell markers (both CD3 and CD45RO) negative. The tumors can be diagnosed as human B-cell lymphomas by these morphological and immunohistochemical features. In situ hybridization exhibited resultant tumor cells had EBV encoded small RNA-1 (EBER-1). Human-derived IgG could be found in the serum from SCID mice on the 15th day following hu-PBL transplantation, and IgG levels increased with the tumor development in 6 hu-PBL/SCID chimeras. Intraperitoneal transfer of hu-PBLs from EBV+ donors to SCID mice leads to high human IgG levels in mouse serum and B cell lymphomas. Our findings suggest that increasing levels of human-derived IgG in peripheral blood from hu-PBL/SCID mice could be used to monitor EBV-related human B-cell lymphoma development in experimental animals.

  5. Hexachlorophene suppresses beta-catenin expression by up-regulation of Siah-1 in EBV-infected B lymphoma cells.

    Science.gov (United States)

    Min, Hye-Jin; Cho, Il-Rae; Srisuttee, Ratakorn; Park, Eun-Hee; Cho, Dae Ho; Ahn, Jin-Hyun; Lee, Im-Soon; Johnston, Randal N; Oh, Sangtaek; Chung, Young-Hwa

    2009-04-18

    Many studies have shown that the activation of beta-catenin signaling can promote oncogenesis, and it is therefore of interest to find agents that modulate this pathway. Recent work has shown using B lymphoma cells that infection by Epstein-Barr virus (EBV) and expression of its latent membrane protein (LMP)-1, cause increases in the expression of beta-catenin and cellular transformation. Conversely, results from cell-based small molecule screening studies have shown that the antibiotic hexachlorophene can down-regulate beta-catenin in colon cancer cells. Here we report that hexachlorophene also counteracts the elevated beta-catenin levels in EBV-infected B lymphomas. This is associated with restoration in levels of Siah-1 (an E3 ubiquitin ligase that is active in beta-catenin regulation) which had been diminished by LMP-1. Our results suggest that Siah-1 is targeted by both LMP-1 and hexachlorophene with opposite effects. The hexachlorophene modulation of Siah-1 and beta-catenin is independent of p53 and results in reduced expression of cyclin-D1 and c-Myc (target genes of beta-catenin), leading to the growth arrest of B lymphoma cells. From these results we propose that hexachlorophene may provide a novel therapeutic strategy for EBV-infected B lymphoma cells by reducing beta-catenin levels via the restoration of Siah-1.

  6. Characterization of EBV gB indicates properties of both class I and class II viral fusion proteins

    International Nuclear Information System (INIS)

    Backovic, Marija; Leser, George P.; Lamb, Robert A.; Longnecker, Richard; Jardetzky, Theodore S.

    2007-01-01

    To gain insight into Epstein-Barr virus (EBV) glycoprotein B (gB), recombinant, secreted variants were generated. The role of putative transmembrane regions, the proteolytic processing and the oligomerization state of the gB variants were investigated. Constructs containing 2 of 3 C-terminal hydrophobic regions were secreted, indicating that these do not act as transmembrane anchors. The efficiency of cleavage of the gB furin site was found to depend on the nature of C-terminus. All of the gB constructs formed rosette structures reminiscent of the postfusion aggregates formed by other viral fusion proteins. However, substitution of putative fusion loop residues, WY 112-113 and WLIY 193-196 , with less hydrophobic amino acids from HSV-1 gB, produced trimeric protein and abrogated the ability of the EBV gB ectodomains to form rosettes. These data demonstrate biochemical features of EBV gB that are characteristic of other class I and class II viral fusion proteins, but not of HSV-1 gB

  7. Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α

    Science.gov (United States)

    Anyndita, Nadya V. M.; Dluha, Nurul; Himmah, Karimatul; Rifa'i, Muhaimin; Widodo

    2017-11-01

    Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn't give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.

  8. Detection and quantification of Epstein-Barr virus EBER1 in EBV-infected cells by fluorescent in situ hybridization and flow cytometry

    Science.gov (United States)

    Stowe, R. P.; Cubbage, M. L.; Sams, C. F.; Pierson, D. L.; Barrett, A. D.

    1998-01-01

    A rapid and highly sensitive fluorescent in situ hybridization (FISH) assay was developed to detect Epstein Barr virus (EBV)-infected cells in peripheral blood. Multiple fluorescein-labeled antisense oligonucleotide probes were designed to hybridize to the EBER1 transcript, which is highly expressed in latently infected cells. After a rapid (30 min) hybridization, the cells were analyzed by flow cytometry. EBER1 was detected in several positive control cell lines that have variable numbers of EBV genome copies. No EBER1 was detected in two known EBV-negative cell lines. Northern blot analyses confirmed the presence and quantity of EBER1 transcripts in each cell line. This method was used to quantify the number of EBV-infected cells in peripheral blood from a patient with chronic mononucleosis. These results indicate that EBV-infected cells can be detected at the single cell level, and that this assay can be used to quantify the number of EBV-infected cells in clinical samples.

  9. Gemcitabine and capecitabine for heavily pre-treated metastatic colorectal cancer patients

    DEFF Research Database (Denmark)

    Spindler, Karen-Lise G; Pallisgaard, Niels; Andersen, Rikke F

    2014-01-01

    AIM: We investigated the efficacy and safety of capecitabine and gemcitabin (GemCap) in heavily pre-treated, therapy-resistant metastatic colorectal cancer (mCRC) patients and the clinical importance of cell-free DNA (cfDNA) measurement. PATIENTS AND METHODS: Patients' inclusion criteria included...... histopathologically-verified mCRC refractory to standard chemotherapy, adequate organ function and performance status. Treatment included capecitabine (2,000 mg/m(2) day on days 1-7 q2w) and gemcitabine (1,000 mg/m(2) on day 1). The number of DNA alleles was measured in pre-treatment plasma samples using an in...

  10. Somatic cell hybrids between human lymphoma lines. II. Spontaneous and induced patterns of the Epstein-Barr virus (EBV) cycle.

    Science.gov (United States)

    Klein, G; Clements, G; Zeuthen, J; Westman, A

    1976-06-15

    The regulation of spontaneous, IUDR-induced and P3HR-1 virus-induced EA and VCA production patterns was studied in two new somatic hybrids between human lymphoma lines. The hybrid 8A was derived from the crossing of the non-producer Raji with the spontaneous producer Daudi line. The second hybrid, 83, was produced by the fusion of Raji with the EBV-genome-negative B-lymphoma line, BJAB. The studies suggest the following EBV regulation patterns: (1) the spontaneous production of EA and VCA appears to be regulated by controls that differ from the regulators of P3HR-1 virus-induced or IUDR-induced EA synthesis. While spontaneous producer status was dominant over non-producer status, the level of EA inducibility was set by one of the parental cells, Raji ATG, and could either raise (in the previously studied Raji/Namalwa hybrid, cf Nyormoi et al. 1973) or depress (in Raji/Daudi) the level of relative EA inducibility found in the partner cell. (2) Although EA production is a prerequisite for VCA synthesis, the latter is under its own restriction mechanisms, quite independent of those that regulate the level of EA synthesis. (3) Inducibility of EA synthesis by P3HR-1 virus and by IUDR appear to be under the influence of at least partially identical controls. (4) EBV-negative lymphoma cells, exemplified by BJAB, may exert a "complementation" effect on the EA inducibility of their EBV-positive fusion partner, in spite of their own restrictivity against virus-induced EA synthesis. In more general terms, it is obvious that the EBV cycle is under the influence of multiple regulatory mechanisms in the human lymphoid cell. Depending on the parental cell and viral genomes that are allowed to interact, somatic cell hybrids may display a variety of patterns. At this time, cell hybridization is one of the few pathways that permit an approach to this complex and completely unknown world.

  11. Processes for pretreating lignocellulosic biomass: A review

    Energy Technology Data Exchange (ETDEWEB)

    McMillan, J.D.

    1992-11-01

    This paper reviews existing and proposed pretreatment processes for biomass. The focus is on the mechanisms by which the various pretreatments act and the influence of biomass structure and composition on the efficacy of particular pretreatment techniques. This analysis is used to identify pretreatment technologies and issues that warrant further research.

  12. Human B cells fail to secrete type I interferons upon cytoplasmic DNA exposure.

    Science.gov (United States)

    Gram, Anna M; Sun, Chenglong; Landman, Sanne L; Oosenbrug, Timo; Koppejan, Hester J; Kwakkenbos, Mark J; Hoeben, Rob C; Paludan, Søren R; Ressing, Maaike E

    2017-11-01

    Most cells are believed to be capable of producing type I interferons (IFN I) as part of an innate immune response against, for instance, viral infections. In macrophages, IFN I is potently induced upon cytoplasmic exposure to foreign nucleic acids. Infection of these cells with herpesviruses leads to triggering of the DNA sensors interferon-inducible protein 16 (IFI16) and cyclic GMP-AMP (cGAMP) synthase (cGAS). Thereby, the stimulator of interferon genes (STING) and the downstream molecules TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3) are sequentially activated culminating in IFN I secretion. Human gamma-herpesviruses, such as Epstein-Barr virus (EBV), exploit B cells as a reservoir for persistent infection. In this study, we investigated whether human B cells, similar to macrophages, engage the cytoplasmic DNA sensing pathway to induce an innate immune response. We found that the B cells fail to secrete IFN I upon cytoplasmic DNA exposure, although they express the DNA sensors cGAS and IFI16 and the signaling components TBK1 and IRF3. In primary human B lymphocytes and EBV-negative B cell lines, this deficiency is explained by a lack of detectable levels of the central adaptor protein STING. In contrast, EBV-transformed B cell lines did express STING, yet both these lines as well as STING-reconstituted EBV-negative B cells did not produce IFN I upon dsDNA or cGAMP stimulation. Our combined data show that the cytoplasmic DNA sensing pathway is dysfunctional in human B cells. This exemplifies that certain cell types cannot induce IFN I in response to cytoplasmic DNA exposure providing a potential niche for viral persistence. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  13. The effect of heat pretreatment temperature on fermentative hydrogen production using mixed cultures

    Energy Technology Data Exchange (ETDEWEB)

    Baghchehsaraee, Bita; Nakhla, George; Karamanev, Dimitre; Margaritis, Argyrios [Department of Chemical and Biochemical Engineering, The University of Western Ontario, London, Ontario N6A 5B9 (Canada); Reid, Gregor [Department of Microbiology and Immunology, The University of Western Ontario, London, Ontario (Canada); Canadian Research and Development Center for Probiotics, Lawson Health Research Institute, 268 Grosvenor Street, London, Ontario N6A 4V2 (Canada)

    2008-08-15

    The effect of heat treatment at different temperatures on two types of inocula, activated sludge and anaerobically digested sludge, was investigated in batch cultures. Heat treatments were conducted at 65, 80 and 95 C for 30 min. The untreated inocula produced less amount of hydrogen than the pretreated inocula, with lactic acid as the main metabolite. The maximum yields of 2.3 and 1.6 mol H{sub 2}/mol glucose were achieved for the 65 C pretreated anaerobically digested and activated sludges, respectively. Approximately a 15% decrease in yield was observed with increasing pretreatment temperature from 65 to 95 C concomitant with an increase in butyrate/acetate ratio from 1.5 to 2.4 for anaerobically digested sludge. The increase of pretreatment temperature of activated sludge to 95 C suppressed the hydrogen production by lactic acid fermentation. DNA analysis of the microbial community showed that the elevated pretreatment temperatures reduced the species diversity. (author)

  14. T-cell receptor (TCR) phenotype of nodal Epstein-Barr virus (EBV)-positive cytotoxic T-cell lymphoma (CTL): a clinicopathologic study of 39 cases.

    Science.gov (United States)

    Kato, Seiichi; Asano, Naoko; Miyata-Takata, Tomoko; Takata, Katsuyoshi; Elsayed, Ahmed Ali; Satou, Akira; Takahashi, Emiko; Kinoshita, Tomohiro; Nakamura, Shigeo

    2015-04-01

    Among Epstein-Barr virus (EBV)-positive cytotoxic T/NK-cell lymphoma, there are only a few reports on the clinicopathologic features of patients with primary nodal presentation (nodal EBV cytotoxic T-cell lymphoma [CTL]). Here, we compared the clinicopathologic profiles of 39 patients with nodal EBV CTL with those of 27 cases of "extranasal" NK/T-cell lymphoma of nasal type (ENKTL), especially addressing their T-cell receptor (TCR) phenotype. Histologically, 22 of 39 nodal EBV CTL cases (56%) were unique in having centroblastoid appearance, which was contrasted with the lower incidence of this feature in ENKTL (15%, P=0.001). In contrast, pleomorphic appearance was more frequently seen in ENKTL than in nodal EBV CTL (67% vs. 23%, P=0.001). Thirty-three of 39 nodal EBV CTL cases (85%) were of T-cell lineage on the basis of TCR expression and/or TCRγ gene rearrangement; in detail, 18 cases (46%) were TCRβ positive (αβ T), 5 (13%) were TCRγ and/or δ positive (γδ T), and 10 (26%) were TCR-silent type with clonal TCRγ gene rearrangement but no expression of TCRβ, γ, or δ. These results were clearly contrasted by a lower incidence of T-cell lineage in ENKTL (7 cases, 26%, P<0.001). Notably, the survival time of the 5 nodal lymphoma patients with γδ T-cell phenotype was within 3 months, which was inferior to those of αβ T and TCR-silent types (P=0.003), and 3 of those with available clinical information were all found to be associated with autoimmune diseases. These data suggest that nodal EBV CTL is distinct from ENKTL.

  15. Similarities between the Epstein-Barr Virus (EBV) Nuclear Protein EBNA1 and the Pioneer Transcription Factor FoxA: Is EBNA1 a “Bookmarking” Oncoprotein that Alters the Host Cell Epigenotype?

    Science.gov (United States)

    Niller, Hans Helmut; Minarovits, Janos

    2012-01-01

    EBNA1, a nuclear protein expressed in all EBV-associated neoplasms is indispensable for the maintenance of the viral episomes in latently infected cells. EBNA1 may induce genetic alterations by upregulating cellular recombinases, production of reactive oxygen species (ROS) and affecting p53 levels and function. All these changes may contribute to tumorigenesis. In this overview we focus, however, on the epigenetic alterations elicited by EBNA1 by drawing a parallel between EBNA1 and the FoxA family of pioneer transcription factors. Both EBNA1 and FoxA induce local DNA demethylation, nucleosome destabilization and bind to mitotic chromosomes. Local DNA demethylation and nucleosome rearrangement mark active promoters and enhancers. In addition, EBNA1 and FoxA, when associated with mitotic chromatin may “bookmark” active genes and ensure their reactivation in postmitotic cells (epigenetic memory). We speculate that DNA looping induced by EBNA1-EBNA1 interactions may reorganize the cellular genome. Such chromatin loops, sustained in mitotic chromatin similarly to the long-distance interactions mediated by the insulator protein CTCF, may also mediate the epigenetic inheritance of gene expression patterns. We suggest that EBNA1 has the potential to induce patho-epigenetic alterations contributing to tumorigenesis. PMID:25436603

  16. Similarities between the Epstein-Barr Virus (EBV Nuclear Protein EBNA1 and the Pioneer Transcription Factor FoxA: Is EBNA1 a “Bookmarking” Oncoprotein that Alters the Host Cell Epigenotype?

    Directory of Open Access Journals (Sweden)

    Hans Helmut Niller

    2012-09-01

    Full Text Available EBNA1, a nuclear protein expressed in all EBV-associated neoplasms is indispensable for the maintenance of the viral episomes in latently infected cells. EBNA1 may induce genetic alterations by upregulating cellular recombinases, production of reactive oxygen species (ROS and affecting p53 levels and function. All these changes may contribute to tumorigenesis. In this overview we focus, however, on the epigenetic alterations elicited by EBNA1 by drawing a parallel between EBNA1 and the FoxA family of pioneer transcription factors. Both EBNA1 and FoxA induce local DNA demethylation, nucleosome destabilization and bind to mitotic chromosomes. Local DNA demethylation and nucleosome rearrangement mark active promoters and enhancers. In addition, EBNA1 and FoxA, when associated with mitotic chromatin may “bookmark” active genes and ensure their reactivation in postmitotic cells (epigenetic memory. We speculate that DNA looping induced by EBNA1-EBNA1 interactions may reorganize the cellular genome. Such chromatin loops, sustained in mitotic chromatin similarly to the long-distance interactions mediated by the insulator protein CTCF, may also mediate the epigenetic inheritance of gene expression patterns. We suggest that EBNA1 has the potential to induce patho-epigenetic alterations contributing to tumorigenesis.

  17. Similarities between the Epstein-Barr Virus (EBV) Nuclear Protein EBNA1 and the Pioneer Transcription Factor FoxA: Is EBNA1 a "Bookmarking" Oncoprotein that Alters the Host Cell Epigenotype?

    Science.gov (United States)

    Niller, Hans Helmut; Minarovits, Janos

    2012-09-17

    EBNA1, a nuclear protein expressed in all EBV-associated neoplasms is indispensable for the maintenance of the viral episomes in latently infected cells. EBNA1 may induce genetic alterations by upregulating cellular recombinases, production of reactive oxygen species (ROS) and affecting p53 levels and function. All these changes may contribute to tumorigenesis. In this overview we focus, however, on the epigenetic alterations elicited by EBNA1 by drawing a parallel between EBNA1 and the FoxA family of pioneer transcription factors. Both EBNA1 and FoxA induce local DNA demethylation, nucleosome destabilization and bind to mitotic chromosomes. Local DNA demethylation and nucleosome rearrangement mark active promoters and enhancers. In addition, EBNA1 and FoxA, when associated with mitotic chromatin may "bookmark" active genes and ensure their reactivation in postmitotic cells (epigenetic memory). We speculate that DNA looping induced by EBNA1-EBNA1 interactions may reorganize the cellular genome. Such chromatin loops, sustained in mitotic chromatin similarly to the long-distance interactions mediated by the insulator protein CTCF, may also mediate the epigenetic inheritance of gene expression patterns. We suggest that EBNA1 has the potential to induce patho-epigenetic alterations contributing to tumorigenesis.

  18. Tank Focus Area pretreatment activities

    International Nuclear Information System (INIS)

    McGinnis, C.P.; Welch, T.D.; Manke, K.L.

    1997-01-01

    Plans call for the high-level wastes to be retrieved from the tanks and immobilized in a stable waste form suitable for long-term isolation. Chemistry and chemical engineering operations are required to retrieve the wastes, to condition the wastes for subsequent steps, and to reduce the costs of the waste management enterprise. Pretreatment includes those processes between retrieval and immobilization, and includes preparation of suitable feed material for immobilization and separations to partition the waste into streams that yield lower life-cycle costs. Some of the technologies being developed by the Tank Focus Area (TFA) to process these wastes are described. These technologies fall roughly into three areas: (1) solid/liquid separation (SLS), (2) sludge pretreatment, and (3) supernate pretreatment

  19. Designing and overproducing a tandem epitope of gp350/220 that shows a potential to become an EBV vaccine

    Directory of Open Access Journals (Sweden)

    Widodo

    2018-03-01

    Full Text Available Background: Epstein-Barr virus (EBV can cause cancer in people from around the world. There is no EBV vaccine available for use on a global scale. However, emerging evidence suggests that the epitope on the gp350/220 capsid protein may be developed into an EBV vaccine. Nevertheless, the production of small, single epitope is challenging of stability issues and possible alteration of peptide structure. In this study, a tandem epitope was developed consisting of three single epitopes, aimed to improve stability, antigenicity and preserve epitope structure. Materials and methods: A tandem epitope was designed using bioinformatics based on the epitope structure of the gp350/220 protein. The tandem epitope structure was analyzed using a protein folding method with Abalone software, which was further refined via YASARA force field and molecular repairing using a FoldX method. Immunogenicity was examined with Epitopia software, whereas allergen properties were tested using AlgPred. The pattern of the tandem epitope binding with anti-gp350/220 antibodies was performed using Z-dock and snugDock. The tandem epitope was then overproduced in E. coli strain BL21 as a host cell. Result: Our model demonstrated a successfully designed and overproduced tandem epitope. The tandem epitope demonstrated a similar structure compared with the epitope of whole protein gp350/220. Our epitope also demonstrated non-allergen and antigenicity properties, and possessed antibody binding patterns consistent with whole protein gp350/220. Conclusion and recommendation: These data suggest a novel tandem epitope composed of three similar epitopes demonstrates antigenicity, structure, and binding properties consistent with whole protein gp350/220. We also demonstrate successful production of the tandem epitope using E. coli strain BL21 as a host. Future in vivo experimental animal research is necessary to test the ability of this tandem epitope to stimulate antibody production

  20. Detecção do vírus Epstein-Barr (EBV em adenocarcinomas gástricos procedentes dos estados do Ceará e de São Paulo Epstein-Barr virus (EBV detection in gastric carcinomas from Ceará and São Paulo states, in Brazil

    Directory of Open Access Journals (Sweden)

    Marcos Antonio Pereira de Lima

    2011-04-01

    Full Text Available INTRODUÇÃO: O vírus Epstein-Barr (EBV está associado a cerca de 10% dos adenocarcinomas gástricos, representando mais de 50 mil casos por ano no mundo. Apesar dos estudos realizados em várias partes do mundo, alguns aspectos clinicopatológicos permanecem controversos. OBJETIVOS: O presente estudo teve como objetivo analisar as características clinicopatológicas de casos de adenocarcinomas gástricos procedentes dos estados de São Paulo e Ceará, correlacionando-os com a detecção de EBV. MATERIAIS E MÉTODOS: Foram obtidos 192 casos de adenocarcinomas gástricos de hospitais dos estados de São Paulo e do Ceará, dos quais 160 foram submetidos à técnica de RNA-hibridização in situ para detecção de EBV. RESULTADOS: Dos 160 casos, 11 (6,9% foram EBV-positivo, exibindo intensa marcação nuclear em células tumorais. Destes, dois casos também apresentaram linfócitos infiltrados marcados. Não encontramos marcação em tecido normal ou pré-neoplásico. São Paulo e Ceará apresentaram as frequências 3/60 (5% e 8/100 (8%, respectivamente, e maior relação do EBV com indivíduos do sexo masculino, de idade avançada, com tumores do tipo intestinal, de estadiamento elevado e grau pouco a moderadamente diferenciado. Os casos do Ceará exibiram aumento relativo de tumores EBV(+ localizados na cárdia, enquanto os casos de São Paulo demonstraram aumento naqueles localizados no corpo gástrico. CONCLUSÃO: A frequência de tumores EBV(+ do presente estudo situa-se nos valores descritos na literatura mundial. Entre os achados, um deles não encontra paralelo na literatura mundial e refere-se ao elevado percentual de tumores EBV(+ no corpo gástrico observado nos casos de São Paulo.INTRODUCTION: The Epstein-Barr virus (EBV has been associated with approximately 10% of gastric adenocarcinomas, which represents more than 50,000 cases/year worldwide. Despite the studies undertaken in several countries, some clinical-pathological aspects

  1. Carbonic Acid Pretreatment of Biomass

    Energy Technology Data Exchange (ETDEWEB)

    G. Peter van Walsum; Kemantha Jayawardhana; Damon Yourchisin; Robert McWilliams; Vanessa Castleberry

    2003-05-31

    This project sought to address six objectives, outlined below. The objectives were met through the completion of ten tasks. 1) Solidify the theoretical understanding of the binary CO2/H2O system at reaction temperatures and pressures. The thermodynamics of pH prediction have been improved to include a more rigorous treatment of non-ideal gas phases. However it was found that experimental attempts to confirm theoretical pH predictions were still off by a factor of about 1.8 pH units. Arrhenius experiments were carried out and the activation energy for carbonic acid appears to be substantially similar to sulfuric acid. Titration experiments have not yet confirmed or quantified the buffering or acid suppression effects of carbonic acid on biomass. 2) Modify the carbonic acid pretreatment severity function to include the effect of endogenous acid formation and carbonate buffering, if necessary. It was found that the existing severity functions serve adequately to account for endogenous acid production and carbonate effects. 3) Quantify the production of soluble carbohydrates at different reaction conditions and severity. Results show that carbonic acid has little effect on increasing soluble carbohydrate concentrations for pretreated aspen wood, compared to pretreatment with water alone. This appears to be connected to the release of endogenous acids by the substrate. A less acidic substrate such as corn stover would derive benefit from the use of carbonic acid. 4) Quantify the production of microbial inhibitors at selected reaction conditions and severity. It was found that the release of inhibitors was correlated to reaction severity and that carbonic acid did not appear to increase or decrease inhibition compared to pretreatment with water alone. 5) Assess the reactivity to enzymatic hydrolysis of material pretreated at selected reaction conditions and severity. Enzymatic hydrolysis rates increased with severity, but no advantage was detected for the use of carbonic

  2. Repositioning antimicrobial agent pentamidine as a disruptor of the lateral interactions of transmembrane domain 5 of EBV latent membrane protein 1.

    Directory of Open Access Journals (Sweden)

    Xiaohui Wang

    Full Text Available The lateral transmembrane protein-protein interactions (PPI have been regarded as "undruggable" despite their importance in many essential biological processes. The homo-trimerization of transmembrane domain 5 (TMD-5 of latent membrane protein 1 (LMP-1 is critical for the constitutive oncogenic activation of the Epstein-Barr virus (EBV. Herein we repurpose the antimicrobial agent pentamidine as a regulator of LMP-1 TMD-5 lateral interactions. The results of ToxR assay, tryptophan fluorescence assay, courmarin fluorescence dequenching assay, and Bis-Tris sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE consistently show pentamidine disrupts LMP-1 TMD-5 lateral interactions. Furthermore, pentamidine inhibits LMP-1 signaling, inducing cellular apoptosis and suppressing cell proliferation in the EBV infected B cells. In contrast, EBV negative cells are less susceptible to pentamidine. This study provides a novel non-peptide small molecule agent for regulating LMP-1 TMD-5 lateral interactions.

  3. ASSOCIATION OF EPSTEIN-BARR VIRUS (EBV) BUT NOT HUMAN PAPILLOMAVIRUS (HPV) WITH GINGIVITIS AND/OR PERIODONTITIS IN TRANSPLANTED INDIVIDUALS.

    Science.gov (United States)

    Baez, Camila Freze; Savassi-Ribas, Flavia; Rocha, Wilker Menezes da; Almeida, Stéphanie G S; Gonçalves, Marianna T V; Guimarães, Maria Angelica A M; Cavalcanti, Silvia Maria B; Varella, Rafael B

    2016-07-11

    The aim of this study was to investigate the association of EBV and HPV with gingivitis and/or periodontitis according to the immunologic status. To this end, 74 oral biopsies from transplanted and non-transplanted individuals with the abovementioned oral manifestations were submitted to a screening by PCR for both viruses. According to the results, EBV was strongly associated with gingivitis and/or periodontitis in transplanted individuals (p = 0.011) but not HPV (p = 0.766). EBV-HPV co-detections did not enhance the presence of tissue injury as well. Although a causal relationship was not investigated in this study, the higher frequency of these two oncoviruses in lesion tissues must be investigated in follow-up studies, especially among immunocompromised individuals.

  4. ASSOCIATION OF EPSTEIN-BARR VIRUS (EBV BUT NOT HUMAN PAPILLOMAVIRUS (HPV WITH GINGIVITIS AND/OR PERIODONTITIS IN TRANSPLANTED INDIVIDUALS

    Directory of Open Access Journals (Sweden)

    Camila Freze BAEZ

    2016-01-01

    Full Text Available SUMMARY The aim of this study was to investigate the association of EBV and HPV with gingivitis and/or periodontitis according to the immunologic status. To this end, 74 oral biopsies from transplanted and non-transplanted individuals with the abovementioned oral manifestations were submitted to a screening by PCR for both viruses. According to the results, EBV was strongly associated with gingivitis and/or periodontitis in transplanted individuals (p = 0.011 but not HPV (p = 0.766. EBV-HPV co-detections did not enhance the presence of tissue injury as well. Although a causal relationship was not investigated in this study, the higher frequency of these two oncoviruses in lesion tissues must be investigated in follow-up studies, especially among immunocompromised individuals.

  5. Ancient DNA

    DEFF Research Database (Denmark)

    Willerslev, Eske; Cooper, Alan

    2004-01-01

    ancient DNA, palaeontology, palaeoecology, archaeology, population genetics, DNA damage and repair......ancient DNA, palaeontology, palaeoecology, archaeology, population genetics, DNA damage and repair...

  6. Novel method for detection of virus-specific CD41+ T cells indicates a decreased EBV-specific CD4+ T cell response in untreated HIV-infected subjects

    NARCIS (Netherlands)

    Piriou, Erwan R.; van Dort, Karel; Nanlohy, Nening M.; van Oers, Marinus H.; Miedema, Frank; van Baarle, Debbie

    2005-01-01

    A lower function of EBV-specific CD8(+) T cells in HIV-infected subjects could be related to a lack of specific CD4(+) T cell help. Therefore, we studied EBV-specific CD4(+) T cells in both healthy donors and untreated or highly active antiretroviral therapy (HAART)-treated HIV-seropositive

  7. Pretreatment Solution for Water Recovery Systems

    Science.gov (United States)

    Muirhead, Dean (Inventor)

    2018-01-01

    Chemical pretreatments are used to produce usable water by treating a water source with a chemical pretreatment that contains a hexavalent chromium and an acid to generate a treated water source, wherein the concentration of sulfate compounds in the acid is negligible, and wherein the treated water source remains substantially free of precipitates after the addition of the chemical pretreatment. Other methods include reducing the pH in urine to be distilled for potable water extraction by pretreating the urine before distillation with a pretreatment solution comprising one or more acid sources selected from a group consisting of phosphoric acid, hydrochloric acid, and nitric acid, wherein the urine remains substantially precipitate free after the addition of the pretreatment solution. Another method described comprises a process for reducing precipitation in urine to be processed for water extraction by mixing the urine with a pretreatment solution comprising hexavalent chromium compound and phosphoric acid.

  8. Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities

    Science.gov (United States)

    Fei Cheng; Lin Hou; Keith Woeste; Zhengchun Shang; Xiaobang Peng; Peng Zhao; Shuoxin Zhang

    2016-01-01

    Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high...

  9. A soluble form of Epstein-Barr virus gH/gL inhibits EBV-induced membrane fusion and does not function in fusion

    Energy Technology Data Exchange (ETDEWEB)

    Rowe, Cynthia L. [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Connolly, Sarah A. [Department of Health Sciences, DePaul University, Chicago, IL 60614 (United States); Chen, Jia [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States); Jardetzky, Theodore S. [Department of Structural Biology, Stanford University School of Medicine, 371 Serra Mall, Stanford, CA 94305 (United States); Longnecker, Richard, E-mail: r-longnecker@northwestern.edu [Department of Microbiology and Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 (United States)

    2013-02-05

    We investigated whether soluble EBV gH/gL (sgH/gL) functions in fusion and made a series of truncations of gH/gL domains based on the gH/gL crystal structure. We found sgH/gL failed to mediate cell-cell fusion both when co-expressed with the other entry glycoproteins and when added exogenously to fusion assays. Interestingly, sgH/gL inhibited cell-cell fusion in a dose dependent manner when co-expressed. sgH/gL from HSV was unable to inhibit EBV fusion, suggesting the inhibition was specific to EBV gH/gL. sgH/gL stably binds gp42, but not gB nor gH/gL. The domain mutants, DI/gL, DI-II/gL and DI-II-III/gL were unable to bind gp42. Instead, DI-II/gL, DI-II-III/gL and sgH/gL but not DI/gL decreased the expression of gp42, resulting in decreased overall fusion. Overall, our results suggest that domain IV may be required for proper folding and the transmembrane domain and cytoplasmic tail of EBV gH/gL are required for the most efficient fusion.

  10. In Vitro and In Vivo Antitumor Effect of Anti-CD33 Chimeric Receptor-Expressing EBV-CTL against CD33+ Acute Myeloid Leukemia

    Directory of Open Access Journals (Sweden)

    A. Dutour

    2012-01-01

    Full Text Available Genetic engineering of T cells with chimeric T-cell receptors (CARs is an attractive strategy to treat malignancies. It extends the range of antigens for adoptive T-cell immunotherapy, and major mechanisms of tumor escape are bypassed. With this strategy we redirected immune responses towards the CD33 antigen to target acute myeloid leukemia. To improve in vivo T-cell persistence, we modified human Epstein Barr Virus-(EBV- specific cytotoxic T cells with an anti-CD33.CAR. Genetically modified T cells displayed EBV and HLA-unrestricted CD33 bispecificity in vitro. In addition, though showing a myeloablative activity, they did not irreversibly impair the clonogenic potential of normal CD34+ hematopoietic progenitors. Moreover, after intravenous administration into CD33+ human acute myeloid leukemia-bearing NOD-SCID mice, anti-CD33-EBV-specific T cells reached the tumor sites exerting antitumor activity in vivo. In conclusion, targeting CD33 by CAR-modified EBV-specific T cells may provide additional therapeutic benefit to AML patients as compared to conventional chemotherapy or transplantation regimens alone.

  11. PiggyBac-mediated cancer immunotherapy using EBV-specific cytotoxic T-cells expressing HER2-specific chimeric antigen receptor.

    Science.gov (United States)

    Nakazawa, Yozo; Huye, Leslie E; Salsman, Vita S; Leen, Ann M; Ahmed, Nabil; Rollins, Lisa; Dotti, Gianpietro; Gottschalk, Stephen M; Wilson, Matthew H; Rooney, Cliona M

    2011-12-01

    Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTLs) can be modified to function as heterologous tumor directed effector cells that survive longer in vivo than tumor directed T cells without virus specificity, due to chronic stimulation by viral antigens expressed during persistent infection in seropositive individuals. We evaluated the nonviral piggyBac (PB) transposon system as a platform for modifying EBV-CTLs to express a functional human epidermal growth factor receptor 2-specific chimeric antigen receptor (HER2-CAR) thereby directing virus-specific, gene modified CTLs towards HER2-positive cancer cells. Peripheral blood mononuclear cells (PBMCs) were nucleofected with transposons encoding a HER2-CAR and a truncated CD19 molecule for selection followed by specific activation and expansion of EBV-CTLs. HER2-CAR was expressed in ~40% of T cells after CD19 selection with retention of immunophenotype, polyclonality, and function. HER2-CAR-modified EBV-CTLs (HER2-CTLs) killed HER2-positive brain tumor cell lines in vitro, exhibited transient and reversible increases in HER2-CAR expression following antigen-specific stimulation, and stably expressed HER2-CAR beyond 120 days. Adoptive transfer of PB-modified HER2-CTLs resulted in tumor regression in a murine xenograft model. Our results demonstrate that PB can be used to redirect virus-specific CTLs to tumor targets, which should prolong tumor-specific T cell survival in vivo producing more efficacious immunotherapy.

  12. Chronic rhinosinusitis with nasal polyps is characterized by B-cell inflammation and EBV-induced protein 2 expression

    Science.gov (United States)

    Hulse, Kathryn E.; Norton, James E.; Suh, Lydia; Zhong, Qiu; Mahdavinia, Mahboobeh; Simon, Patrick; Kern, Robert C.; Conley, David B.; Chandra, Rakesh K.; Tan, Bruce K.; Peters, Anju T.; Grammer, Leslie C.; Harris, Kathleen E.; Carter, Roderick G.; Kato, Atsushi; Schleimer, Robert P.

    2013-01-01

    Background Despite the high prevalence and morbidity of chronic rhinosinusitis (CRS), little is known about the mechanisms that underlie its pathogenesis. Recent studies have suggested that B cells might play an important role in CRS. Objective We sought to thoroughly characterize B lineage cells within sinus tissues of patients with CRS and healthy control subjects and to determine whether levels of EBV-induced protein 2, which is known to play an important role in the development of B-cell responses, were increased in patients with CRS. Methods Cells isolated from sinus tissues of patients with CRS and healthy control subjects were characterized by means of flow cytometry and immunohistochemistry. Local production of antibodies was measured in tissue extracts, nasal lavage fluid, and sera by using multiplex bead arrays and ELISA. Quantitative RT-PCR, ELISA, and Western blotting were used to assess gene and protein expression from tissue extracts. Results Nasal polyps (NPs) from patients with CRS had increased levels of both B cells and plasma cells compared with uncinate tissue from healthy control subjects (P < .05). NPs also contained significantly increased levels of several antibody isotypes compared with normal uncinate tissue (P < .05), but no differences in circulating antibody levels were found. Interestingly, levels of EBV-induced protein 2 were also increased in NPs (P < .05) and were positively correlated with expression of plasma cell markers (CD138 and B lymphocyte–induced maturation protein) in sinus tissue. Conclusion B cells and plasma cells are enriched in NPs, actively produce antibodies locally, and might contribute to chronic inflammation in patients with CRS. Elucidating the mechanisms that underlie this excessive local B-cell response might provide novel insights for the development of improved therapeutic strategies. PMID:23473835

  13. Replication of Human Herpesviruses Is Associated with Higher HIV DNA Levels during Antiretroviral Therapy Started at Early Phases of HIV Infection

    Science.gov (United States)

    Anderson, Christy M.; Var, Susanna R.; Oliveira, Michelli F.; Lada, Steven M.; Vargas, Milenka V.; Little, Susan J.; Richman, Douglas D.; Strain, Matthew C.; Pérez-Santiago, Josué; Smith, Davey M.

    2016-01-01

    ABSTRACT Asymptomatic replication of human herpesviruses (HHV) is frequent in HIV-infected men and is associated with increased T-cell activation and HIV disease progression. We hypothesized that the presence of replication of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) (the most frequently detected HHV) might influence HIV DNA decay during antiretroviral therapy (ART). We investigated 607 peripheral blood mononuclear cell (PBMC) samples from 107 CMV-seropositive, HIV-infected men who have sex with men, who started ART within a median of 3 months from their estimated date of infection (EDI) and were monitored for a median of 19 months thereafter. Levels of HIV, CMV, and EBV DNA and cellular HIV RNA were measured by droplet digital PCR (ddPCR) for each time point. Using a general linear mixed-effect regression model, we evaluated associations between the presence of detectable CMV DNA and EBV DNA levels and HIV DNA decay and cellular HIV RNA levels, while adjusting for peak HIV RNA, nadir CD4+ count, CD4/CD8 ratio, CMV IgG levels, time from EDI to ART initiation, time from ART initiation to virologic suppression, detectable CMV DNA pre-ART, and age. The presence of intermittent CMV DNA in PBMC during ART was significantly associated with slower decay of HIV DNA (P = 0.011) but not with increased cellular HIV RNA transcription or more detectable 2-long terminal repeat circles. Higher levels of EBV DNA were also associated with higher levels of HIV DNA (P < 0.001) and increased unspliced cellular HIV RNA transcription (P = 0.010). These observations suggest that replication of HHV may help maintain a larger HIV DNA reservoir, but the underlying mechanisms remain unclear. IMPORTANCE Over three-fourths of HIV-infected men have at least one actively replicating human herpesvirus (HHV) in their mucosal secretions at any one time. Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) are the most common, and although it is often asymptomatic, such CMV and EBV

  14. Genetic diversity of EBV-encoded LMP1 in the Swiss HIV Cohort Study and implication for NF-Κb activation.

    Directory of Open Access Journals (Sweden)

    Emilie Zuercher

    Full Text Available Epstein-Barr virus (EBV is associated with several types of cancers including Hodgkin's lymphoma (HL and nasopharyngeal carcinoma (NPC. EBV-encoded latent membrane protein 1 (LMP1, a multifunctional oncoprotein, is a powerful activator of the transcription factor NF-κB, a property that is essential for EBV-transformed lymphoblastoid cell survival. Previous studies reported LMP1 sequence variations and induction of higher NF-κB activation levels compared to the prototype B95-8 LMP1 by some variants. Here we used biopsies of EBV-associated cancers and blood of individuals included in the Swiss HIV Cohort Study (SHCS to analyze LMP1 genetic diversity and impact of sequence variations on LMP1-mediated NF-κB activation potential. We found that a number of variants mediate higher NF-κB activation levels when compared to B95-8 LMP1 and mapped three single polymorphisms responsible for this phenotype: F106Y, I124V and F144I. F106Y was present in all LMP1 isolated in this study and its effect was variant dependent, suggesting that it was modulated by other polymorphisms. The two polymorphisms I124V and F144I were present in distinct phylogenetic groups and were linked with other specific polymorphisms nearby, I152L and D150A/L151I, respectively. The two sets of polymorphisms, I124V/I152L and F144I/D150A/L151I, which were markers of increased NF-κB activation in vitro, were not associated with EBV-associated HL in the SHCS. Taken together these results highlighted the importance of single polymorphisms for the modulation of LMP1 signaling activity and demonstrated that several groups of LMP1 variants, through distinct mutational paths, mediated enhanced NF-κB activation levels compared to B95-8 LMP1.

  15. Constitutive AP-1 activity and EBV infection induce PD-L1 in Hodgkin lymphomas and posttransplant lymphoproliferative disorders: implications for targeted therapy.

    Science.gov (United States)

    Green, Michael R; Rodig, Scott; Juszczynski, Przemyslaw; Ouyang, Jing; Sinha, Papiya; O'Donnell, Evan; Neuberg, Donna; Shipp, Margaret A

    2012-03-15

    Programmed cell death ligand 1 (PD-L1) is a molecule expressed on antigen-presenting cells that engages the PD-1 receptor on T cells and inhibits T-cell receptor signaling. The PD-1 axis can be exploited by tumor cells to dampen host antitumor immune responses and foster tumor cell survival. PD-1 blockade has shown promise in multiple malignancies but should be directed toward patients in whom it will be most effective. In recent studies, we found that the chromosome 9p24.1 amplification increased the gene dosage of PD-L1 and its induction by JAK2 in a subset of patients with classical Hodgkin lymphoma (cHL). However, cHLs with normal 9p24.1 copy numbers also expressed detectable PD-L1, prompting analyses of additional PD-L1 regulatory mechanisms. Herein, we utilized immunohistochemical, genomic, and functional analyses to define alternative mechanisms of PD-L1 activation in cHL and additional EBV(+) lymphoproliferative disorders. We identified an AP-1-responsive enhancer in the PD-L1 gene. In cHL Reed-Sternberg cells, which exhibit constitutive AP-1 activation, the PD-L1 enhancer binds AP-1 components and increases PD-L1 promoter activity. In addition, we defined Epstein-Barr virus (EBV) infection as an alternative mechanism for PD-L1 induction in cHLs with diploid 9p24.1. PD-L1 was also expressed by EBV-transformed lymphoblastoid cell lines as a result of latent membrane protein 1-mediated, JAK/STAT-dependent promoter and AP-1-associated enhancer activity. In addition, more than 70% of EBV(+) posttransplant lymphoproliferative disorders expressed detectable PD-L1. AP-1 signaling and EBV infection represent alternative mechanisms of PD-L1 induction and extend the spectrum of tumors in which to consider PD-1 blockade.

  16. Associations between Burkitt lymphoma among children in Malawi and infection with HIV, EBV and malaria: results from a case-control study.

    Directory of Open Access Journals (Sweden)

    Nora Mutalima

    Full Text Available BACKGROUND: Burkitt lymphoma, a childhood cancer common in parts of sub-Saharan Africa, has been associated with Epstein Barr Virus (EBV and malaria, but its association with human immunodeficiency virus (HIV is not clear. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a case-control study of Burkitt lymphoma among children (aged < or = 15 years admitted to the pediatric oncology unit in Blantyre, Malawi between July 2005 and July 2006. Cases were 148 children diagnosed with Burkitt lymphoma and controls were 104 children admitted with non-malignant conditions or cancers other than hematological malignancies and Kaposi sarcoma. Interviews were conducted and serological samples tested for antibodies against HIV, EBV and malaria. Odds ratios for Burkitt lymphoma were estimated using unconditional logistic regression adjusting for sex, age, and residential district. Cases had a mean age of 7.1 years and 60% were male. Cases were more likely than controls to be HIV positive (Odds ratio (OR = 12.4, 95% Confidence Interval (CI 1.3 to 116.2, p = 0.03. ORs for Burkitt lymphoma increased with increasing antibody titers against EBV (p = 0.001 and malaria (p = 0.01. Among HIV negative participants, cases were thirteen times more likely than controls to have raised levels of both EBV and malaria antibodies (OR = 13.2; 95% CI 3.8 to 46.6; p = 0.001. Reported use of mosquito nets was associated with a lower risk of Burkitt lymphoma (OR = 0.2, 95% CI, 0.03 to 0.9, p = 0.04. CONCLUSIONS: Our findings support prior evidence that EBV and malaria act jointly in the pathogenesis of Burkitt lymphoma, suggesting that malaria prevention may decrease the risk of Burkitt lymphoma. HIV may also play a role in the etiology of this childhood tumor.

  17. Method for pretreating lignocellulosic biomass

    Science.gov (United States)

    Kuzhiyil, Najeeb M.; Brown, Robert C.; Dalluge, Dustin Lee

    2015-08-18

    The present invention relates to a method for pretreating lignocellulosic biomass containing alkali and/or alkaline earth metal (AAEM). The method comprises providing a lignocellulosic biomass containing AAEM; determining the amount of the AAEM present in the lignocellulosic biomass; identifying, based on said determining, the amount of a mineral acid sufficient to completely convert the AAEM in the lignocellulosic biomass to thermally-stable, catalytically-inert salts; and treating the lignocellulosic biomass with the identified amount of the mineral acid, wherein the treated lignocellulosic biomass contains thermally-stable, catalytically inert AAEM salts.

  18. Novel Epstein-Barr virus-like particles incorporating gH/gL-EBNA1 or gB-LMP2 induce high neutralizing antibody titers and EBV-specific T-cell responses in immunized mice.

    Science.gov (United States)

    Perez, Elizabeth M; Foley, Joslyn; Tison, Timelia; Silva, Rute; Ogembo, Javier Gordon

    2017-03-21

    Previous Epstein-Barr virus (EBV) prophylactic vaccines based on the major surface glycoprotein gp350/220 as an immunogen have failed to block viral infection in humans, suggesting a need to target other viral envelope glycoproteins. In this study, we reasoned that incorporating gH/gL or gB, critical glycoproteins for viral fusion and entry, on the surface of a virus-like particle (VLP) would be more immunogenic than gp350/220 for generating effective neutralizing antibodies to prevent viral infection of both epithelial and B cell lines. To boost the humoral response and trigger cell-mediated immunity, EBV nuclear antigen 1 (EBNA1) and latent membrane protein 2 (LMP2), intracellular latency proteins expressed in all EBV-infected cells, were also included as critical components of the polyvalent EBV VLP. gH/gL-EBNA1 and gB-LMP2 VLPs were efficiently produced in Chinese hamster ovary cells, an FDA-approved vehicle for mass-production of biologics. Immunization with gH/gL-EBNA1 and gB-LMP2 VLPs without adjuvant generated both high neutralizing antibody titers in vitro and EBV-specific T-cell responses in BALB/c mice. These data demonstrate that will be invaluable not only in preventing EBV infection, but importantly, in preventing and treating the 200,000 cases of EBV-associated cancers that occur globally every year.

  19. Radio response of human lymphocytes pretreated with boron and gadoliniums assessed by the, comet assay

    International Nuclear Information System (INIS)

    Kim, J. K.; Park, T. W.; Cebulska-Wasiewska, A.; Nili, M.

    2009-01-01

    Boron and gadolinium are among the nuclides that hold a unique property of being a neutron capture therapy agent. Neutron beams have often a considerable portion of gamma rays with fast neutrons. Gamma rays, as beam contaminants, can cause considerable damage to normal tissues even if such tissues do contain high boron concentrations. Materials and Methods: The modification of radio response in human lymphocytes pretreated with boron or gadolinium compound was studied by assessing the DNA damage using single cell gel electrophoresis, the comet assay. The lymphocytes from the human peripheral blood were irradiated with 0, 1, 2 and 4 Gy of gamma rays from a 60 Co isotopic source with or without pretreatment of boron or gadolinium compound for 10 minutes at 4 d egree C . Post-irradiation procedures included slide preparation, cell-lysing, unwinding and electrophoresis, neutralization, staining, and analytic steps, gel electrophoresis. Results: The results indicate that pretreatment with boron compound (50 n M or 250 n M of 10 B) is effective in reducing the radiosensitivity of the lymphocyte DNA. Conversely, pretreatment with gadolinium compound (50 n M) led to a dose-dependent increase in the radiosensitivity, most prominently with a dose of 4 Gy (P<0.001). Furthermore, when the lymphocytes were pretreated with a Combined mixture (1:1) of boron (250 n M) and gadolinium (50 n M) compounds, the reduced radiosensitivity was also observed.

  20. Wash water waste pretreatment system

    Science.gov (United States)

    1977-01-01

    Investigations were completed on wash waters based on each candidate personal cleansing agent. Evaluations of coagulants, antifoam agents, and the effect of promising antifoams on the chemical precipitation were included. Based on these evaluations two candidate soaps as well as their companion antifoam agents were selected for further work. Operating parameters included the effect of soap concentration, ferric chloride concentration, duration of mixing, and pore size of depth filters on the degree of soap removal. The effect of pressure on water flow through filter cartridges and on the rate of decline of water flow was also investigated. The culmination of the program was the recommendation of a pretreatment concept based on chemical precipitation followed by pressure filtration.

  1. Pretreatment of the macroalgae Chaetomorpha linum for the production of bioethanol - Comparison of five pretreatment technologies

    DEFF Research Database (Denmark)

    Schultz-Jensen, Nadja; Thygesen, Anders; Thomsen, Sune Tjalfe

    2013-01-01

    -assisted pretreatment (PAP) and ball milling (BM), to determine effects of the pretreatment methods on the conversion of C. linum into ethanol by simultaneous saccharification and fermentation (SSF). WO and BM showed the highest ethanol yield of 44. g ethanol/100. g glucan, which was close to the theoretical ethanol......A qualified estimate for pretreatment of the macroalgae Chaetomorpha linum for ethanol production was given, based on the experience of pretreatment of land-based biomass. C. linum was subjected to hydrothermal pretreatment (HTT), wet oxidation (WO), steam explosion (STEX), plasma...... yield of 57. g ethanol/100. g glucan. A 64% higher ethanol yield, based on raw material, was reached after pretreatment with WO and BM compared with unpretreated C. linum, however 50% of the biomass was lost during WO. Results indicated that the right combination of pretreatment and marine macroalgae...

  2. Oncogenic Viruses and Breast Cancer: Mouse Mammary Tumor Virus (MMTV), Bovine Leukemia Virus (BLV), Human Papilloma Virus (HPV), and Epstein–Barr Virus (EBV)

    OpenAIRE

    James S. Lawson; Brian Salmons; Wendy K. Glenn

    2018-01-01

    BackgroundAlthough the risk factors for breast cancer are well established, namely female gender, early menarche and late menopause plus the protective influence of early pregnancy, the underlying causes of breast cancer remain unknown. The development of substantial recent evidence indicates that a handful of viruses may have a role in breast cancer. These viruses are mouse mammary tumor virus (MMTV), bovine leukemia virus (BLV), human papilloma viruses (HPVs), and Epstein–Barr virus (EBV-al...

  3. The Definition of Epstein Barr Virus (EBV)’s Role in HTLV-III Infected USAF Personnel as Related to Disease Progression

    Science.gov (United States)

    1989-04-15

    manifestations. The subgroup of patients with oral hairy leukoplakia (OIIL) had an overall high body burden of EBV, including a high density of (ntinued on...result of the increased longevity of HIV-infected patients through the administration of antiretroviral chemotherapy. Oral hairy leukoplakia was found...Boswell RN: Erhanced Epstein-Barr virus activity in patients with oral hairy leukoplakia and human iMndeficiency virus infection. Presented at the

  4. EBV, HCMV, HHV6, and HHV7 Screening in Bone Marrow Samples from Children with Acute Lymphoblastic Leukemia

    Directory of Open Access Journals (Sweden)

    A. Morales-Sánchez

    2014-01-01

    Full Text Available Acute lymphoblastic leukemia (ALL is the most common cancer in childhood worldwide and Mexico has reported one of the highest incidence rates. An infectious etiology has been suggested and supported by epidemiological evidences; however, the identity of the involved agent(s is not known. We considered that early transmitted lymphotropic herpes viruses were good candidates, since transforming mechanisms have been described for them and some are already associated with human cancers. In this study we interrogated the direct role of EBV, HCMV, HHV6, and HHV7 human herpes viruses in childhood ALL. Viral genomes were screened in 70 bone marrow samples from ALL patients through standard and a more sensitive nested PCR. Positive samples were detected only by nested PCR indicating a low level of infection. Our result argues that viral genomes were not present in all leukemic cells, and, hence, infection most likely was not part of the initial genetic lesions leading to ALL. The high statistical power of the study suggested that these agents are not involved in the genesis of ALL in Mexican children. Additional analysis showed that detected infections or coinfections were not associated with prognosis.

  5. Negative interaction between smoking and EBV in the risk of multiple sclerosis: The EnvIMS study.

    Science.gov (United States)

    Bjørnevik, Kjetil; Riise, Trond; Bostrom, Inger; Casetta, Ilaria; Cortese, Marianna; Granieri, Enrico; Holmøy, Trygve; Kampman, Margitta T; Landtblom, Anne-Marie; Magalhaes, Sandra; Pugliatti, Maura; Wolfson, Christina; Myhr, Kjell-Morten

    2017-06-01

    Results from previous studies on a possible interaction between smoking and Epstein-Barr virus (EBV) in the risk of multiple sclerosis (MS) are conflicting. To examine the interaction between smoking and infectious mononucleosis (IM) in the risk of MS. Within the case-control study on Environmental Factors In Multiple Sclerosis (EnvIMS), 1904 MS patients and 3694 population-based frequency-matched healthy controls from Norway, Italy, and Sweden reported on prior exposure to smoking and history of IM. We examined the interaction between the two exposures on the additive and multiplicative scale. Smoking and IM were each found to be associated with an increased MS risk in all three countries, and there was a negative multiplicative interaction between the two exposures in each country separately as well as in the pooled analysis ( p = 0.001). Among those who reported IM, there was no increased risk associated with smoking (odds ratio (OR): 0.95, 95% confidence interval (CI): 0.66-1.37). The direction of the estimated interactions on the additive scale was consistent with a negative interaction in all three countries (relative excess risk due to interaction (RERI): -0.98, 95% CI: -2.05-0.15, p = 0.09). Our findings indicate competing antagonism, where the two exposures compete to affect the outcome.

  6. EBV, HCMV, HHV6, and HHV7 Screening in Bone Marrow Samples from Children with Acute Lymphoblastic Leukemia

    Science.gov (United States)

    Morales-Sánchez, A.; Pompa-Mera, E. N.; Fajardo-Gutiérrez, A.; Alvarez-Rodríguez, F. J.; Bekker-Méndez, V. C.; Flores-Chapa, J. de Diego; Flores-Lujano, J.; Jiménez-Hernández, E.; Peñaloza-González, J. G.; Rodríguez-Zepeda, M. C.; Torres-Nava, J. R.; Velázquez-Aviña, M. M.; Amador-Sánchez, R.; Alvarado-Ibarra, M.; Reyes-Zepeda, N.; Espinosa-Elizondo, R. M.; Pérez-Saldivar, M. L.; Núñez-Enríquez, J. C.; Mejía-Aranguré, J. M.; Fuentes-Pananá, E. M.

    2014-01-01

    Acute lymphoblastic leukemia (ALL) is the most common cancer in childhood worldwide and Mexico has reported one of the highest incidence rates. An infectious etiology has been suggested and supported by epidemiological evidences; however, the identity of the involved agent(s) is not known. We considered that early transmitted lymphotropic herpes viruses were good candidates, since transforming mechanisms have been described for them and some are already associated with human cancers. In this study we interrogated the direct role of EBV, HCMV, HHV6, and HHV7 human herpes viruses in childhood ALL. Viral genomes were screened in 70 bone marrow samples from ALL patients through standard and a more sensitive nested PCR. Positive samples were detected only by nested PCR indicating a low level of infection. Our result argues that viral genomes were not present in all leukemic cells, and, hence, infection most likely was not part of the initial genetic lesions leading to ALL. The high statistical power of the study suggested that these agents are not involved in the genesis of ALL in Mexican children. Additional analysis showed that detected infections or coinfections were not associated with prognosis. PMID:25309913

  7. Integrated analysis of hydrothermal flow through pretreatment

    Directory of Open Access Journals (Sweden)

    Archambault-Leger Veronique

    2012-07-01

    Full Text Available Abstract Background The impact of hydrothermal flowthrough (FT pretreatment severity on pretreatment and solubilization performance metrics was evaluated for three milled feedstocks (corn stover, bagasse, and poplar and two conversion systems (simultaneous saccharification and fermentation using yeast and fungal cellulase, and fermentation by Clostridium thermocellum. Results Compared to batch pretreatment, FT pretreatment consistently resulted in higher XMG recovery, higher removal of non-carbohydrate carbon and higher glucan solubilization by simultaneous saccharification and fermentation (SSF. XMG recovery was above 90% for FT pretreatment below 4.1 severity but decreased at higher severities, particularly for bagasse. Removal of non-carbohydrate carbon during FT pretreatment increased from 65% at low severity to 80% at high severity for corn stover, and from 40% to 70% for bagasse and poplar. Solids obtained by FT pretreatment were amenable to high conversion for all of the feedstocks and conversion systems examined. The optimal time and temperature for FT pretreatment on poplar were found to be 16 min and 210°C. At these conditions, SSF glucan conversion was about 85%, 94% of the XMG was removed, and 62% of the non carbohydrate mass was solubilized. Solubilization of FT-pretreated poplar was compared for C. thermocellum fermentation (10% inoculum, and for yeast-fungal cellulase SSF (5% inoculum, cellulase loading of 5 and 10 FPU/g glucan supplemented with β-glucosidase at 15 and 30 U/g glucan. Under the conditions tested, which featured low solids concentration, C. thermocellum fermentation achieved faster rates and more complete conversion of FT-pretreated poplar than did SSF. Compared to SSF, solubilization by C. thermocellum was 30% higher after 4 days, and was over twice as fast on ball-milled FT-pretreated poplar. Conclusions XMG removal trends were similar between feedstocks whereas glucan conversion trends were significantly

  8. Metallothionein expression during liver regeneration after partial hepatectomy in cadmium-pretreated rats

    Energy Technology Data Exchange (ETDEWEB)

    Margeli, A.P. (Dept. of Forensic Medicine and Toxicology, School of Medicine, Univ. of Athens (Greece)); Theocharis, S.E. (Dept. of Forensic Medicine and Toxicology, School of Medicine, Univ. of Athens (Greece)); Yannacou, N.N. (Dept. of Forensic Medicine and Toxicology, School of Medicine, Univ. of Athens (Greece)); Spiliopoulou, C. (Dept. of Forensic Medicine and Toxicology, School of Medicine, Univ. of Athens (Greece)); Koutselinis, A. (Dept. of Forensic Medicine and Toxicology, School of Medicine, Univ. of Athens (Greece))

    1994-10-01

    Metallothionein is a low molecular mass protein inducible mainly by heavy metals, having high affinity for binding cadmium, zinc and copper. In the present study we investigated the expression of metallothionein in regenerating liver, at different time intervals, in cadmium pretreated partially hepatectomized rats. Liver metallothionein is highly expressed during regeneration induced by partial hepatectomy in rats, providing zinc within the rapidly growing tissue. Cadmium pretreatment caused inhibition of the first peak of liver regeneration, while metallothionein expression was markedly more prominent in the liver residues of cadmium-pretreated rats. These results demonstrate that although metallothionein able to bind temporarily metal ions as zinc and cadmium has been highly expressed, the liver regenerative process was inhibited possibly due to the effects of cadmium on other pivotal events necessary to the DNA replication. (orig.)

  9. Constitutive AP-1 Activity and EBV Infection Induce PD-L1 in Hodgkin Lymphomas and Post-transplant Lymphoproliferative Disorders: Implications for Targeted Therapy

    Science.gov (United States)

    Green, Michael R.; Rodig, Scott; Juszczynski, Przemyslaw; Ouyang, Jing; Sinha, Papiya; O’Donnell, Evan; Neuberg, Donna; Shipp, Margaret A.

    2012-01-01

    Purpose Programmed cell death ligand 1 (PD-L1) is a molecule expressed on antigen-presenting cells that engages the PD-1 receptor on T cells and inhibits T-cell receptor signaling. The PD-1 axis can be exploited by tumor cells to dampen host anti-tumor immune responses and foster tumor cell survival. PD-1 blockade has shown promise in multiple malignancies but should be directed towards patients in whom it will be most effective. In recent studies, we found that the chromosome 9p24.1 amplification increased the gene dosage of PD-L1 and its induction by JAK2 in a subset of patients with classical Hodgkin lymphoma (cHL). However, cHLs with normal 9p24.1 copy numbers also expressed detectable PD-L1, prompting analyses of additional PD-L1 regulatory mechanisms. Experimental Design Herein, we utilized immunohistochemical, genomic and functional analyses to define alternative mechanisms of PD-L1 activation in cHL and additional EBV+ lymphoproliferative disorders. Results We identified an AP-1-responsive enhancer in the PD-L1 gene. In cHL Reed Sternberg cells, which exhibit constitutive AP-1 activation, the PD-L1 enhancer binds AP-1 components and increases PD-L1 promoter activity. In addition, we defined EBV infection as an alternative mechanism for PD-L1 induction in cHLs with diploid 9p24.1. PD-L1 was also expressed by EBV-transformed lymphoblastoid cell lines as a result of latent membrane protein 1-mediated, JAK/STAT-dependent promoter and AP-1-associated enhancer activity. In addition, over 70% of EBV+ post-transplant lymphoproliferative disorders expressed detectable PD-L1. Conclusions AP-1 signaling and EBV infection represent alternative mechanisms of PD-L1 induction and extend the spectrum of tumors in which to consider PD-1 blockade. PMID:22271878

  10. Detection of human papillomavirus and Epstein-Barr virus DNA sequences in oral mucosa of HIV-infected patients by the polymerase chain reaction.

    Science.gov (United States)

    Snijders, P. J.; Schulten, E. A.; Mullink, H.; ten Kate, R. W.; Jiwa, M.; van der Waal, I.; Meijer, C. J.; Walboomers, J. M.

    1990-01-01

    The presence of human papillomavirus (HPV) and Epstein-Barr virus (EBV) was analyzed in 21 oral biopsy specimens of HIV-infected patients using the polymerase chain reaction (PCR) method. Biopsies were categorized as hairy leukoplakia (HL) (n = 12), candidiasis (n = 3), oral warts (n = 2), and clinically normal epithelium (n = 4). For HPV detection a modified general primer-mediated PCR method (GP-PCR), which detects a broad spectrum of HPV genotypes at sub-picogram levels, was used. Human papillomavirus DNA was only found in two oral warts and was identified as HPV type 32. Epstein-Barr virus DNA was detected in 16 biopsy specimens, including the 12 HLs, 2 cases of candidiasis, and 2 samples of normal epithelium. Epstein-Barr virus positivity in HL could be confirmed by Southern blot analysis and DNA in situ hybridization using biotinylated DNA probes (bio-DISH). Epstein-Barr virus bio-DISH was also positive in one sample of normal epithelium from a patient with HL. The results indicate that HL is strongly associated with EBV and not with any of the common HPV types that react with general HPV primers in the PCR. However the detection of EBV in normal oral epithelium by PCR and bio-DISH suggests that the presence of this virus is not exclusively related to HL. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:2169191

  11. The SWI/SNF Chromatin Regulator BRG1 Modulates the Transcriptional Regulatory Activity of the Epstein-Barr Virus DNA Polymerase Processivity Factor BMRF1.

    Science.gov (United States)

    Su, Mei-Tzu; Wang, Ya-Ting; Chen, Yen-Ju; Lin, Su-Fang; Tsai, Ching-Hwa; Chen, Mei-Ru

    2017-05-01

    During the lytic phase of Epstein-Barr virus (EBV), binding of the transactivator Zta to the origin of lytic replication (oriLyt) and the BHLF1 transcript, forming a stable RNA-DNA hybrid, is required to initiate viral DNA replication. EBV-encoded viral DNA replication proteins form complexes to amplify viral DNA. BMRF1, the viral DNA polymerase accessory factor, is essential for lytic DNA replication and also known as a transcriptional regulator of the expression of BHLF1 and BALF2 (single-stranded DNA [ssDNA]-binding protein). In order to determine systematically how BMRF1 regulates viral transcription, a BMRF1 knockout bacmid was generated to analyze viral gene expression using a viral DNA microarray. We found that a subset of Rta-responsive late genes, including BcLF1, BLLF1, BLLF2, and BDLF3, were downregulated in cells harboring a BMRF1 knockout EBV bacmid (p2089ΔBMRF1). In reporter assays, BMRF1 appears to transactivate a subset of viral late promoters through distinct pathways. BMRF1 activates the BDLF3 promoter in an SP1-dependent manner. Notably, BMRF1 associates with the transcriptional regulator BRG1 in EBV-reactivated cells. BMRF1-mediated transactivation activities on the BcLF1 and BLLF1 promoters were attenuated by knockdown of BRG1. In BRG1-depleted EBV-reactivated cells, BcLF1 and BLLF1 transcripts were reduced in number, resulting in reduced virion secretion. BMRF1 and BRG1 bound to the adjacent upstream regions of the BcLF1 and BLLF1 promoters, and depletion of BRG1 attenuated the recruitment of BMRF1 onto both promoters, suggesting that BRG1 is involved in BMRF1-mediated regulation of these two genes. Overall, we reveal a novel pathway by which BMRF1 can regulate viral promoters through interaction with BRG1. IMPORTANCE The cascade of viral gene expression during Epstein-Barr virus (EBV) replication is exquisitely regulated by the coordination of the viral DNA replication machinery and cellular factors. Upon lytic replication, the EBV immediate

  12. Understanding Ionic Liquid Pretreatment of Lignocellulosic Biomasses

    Science.gov (United States)

    Pretreatment of biomass is essential for breaking apart highly ordered and crystalline plant cell walls and loosening the lignin and hemicellulose conjugation to cellulose microfibrills, thereby facilitating enzyme accessibility and adsorption and reducing costs of downstream saccharification proces...

  13. Extrusion Pretreatment of Lignocellulosic Biomass: A Review

    Directory of Open Access Journals (Sweden)

    Jun Zheng

    2014-10-01

    Full Text Available Bioconversion of lignocellulosic biomass to bioethanol has shown environmental, economic and energetic advantages in comparison to bioethanol produced from sugar or starch. However, the pretreatment process for increasing the enzymatic accessibility and improving the digestibility of cellulose is hindered by many physical-chemical, structural and compositional factors, which make these materials difficult to be used as feedstocks for ethanol production. A wide range of pretreatment methods has been developed to alter or remove structural and compositional impediments to (enzymatic hydrolysis over the last few decades; however, only a few of them can be used at commercial scale due to economic feasibility. This paper will give an overview of extrusion pretreatment for bioethanol production with a special focus on twin-screw extruders. An economic assessment of this pretreatment is also discussed to determine its feasibility for future industrial cellulosic ethanol plant designs.

  14. Hydrolysis of alkaline pretreated banana peel

    Science.gov (United States)

    Fatmawati, A.; Gunawan, K. Y.; Hadiwijaya, F. A.

    2017-11-01

    Banana peel is one of food wastes that are rich in carbohydrate. This shows its potential as fermentation substrate including bio-ethanol. This paper presented banana peel alkaline pretreatment and enzymatic hydrolysis. The pretreatment was intended to prepare banana peel in order to increase hydrolysis performance. The alkaline pretreatment used 10, 20, and 30% w/v NaOH solution and was done at 60, 70 and 80°C for 1 hour. The hydrolysis reaction was conducted using two commercial cellulose enzymes. The reaction time was varied for 3, 5, and 7 days. The best condition for pretreatment process was one conducted using 30% NaOH solution and at 80°C. This condition resulted in cellulose content of 90.27% and acid insoluble lignin content of 2.88%. Seven-day hydrolysis time had exhibited the highest reducing sugar concentration, which was7.2869 g/L.

  15. Ultrasonic sludge pretreatment under pressure.

    Science.gov (United States)

    Le, Ngoc Tuan; Julcour-Lebigue, Carine; Delmas, Henri

    2013-09-01

    The objective of this work was to optimize the ultrasound (US) pretreatment of sludge. Three types of sewage sludge were examined: mixed, secondary and secondary after partial methanisation ("digested" sludge). Thereby, several main process parameters were varied separately or simultaneously: stirrer speed, total solid content of sludge (TS), thermal operating conditions (adiabatic vs. isothermal), ultrasonic power input (PUS), specific energy input (ES), and for the first time external pressure. This parametric study was mainly performed for the mixed sludge. Five different TS concentrations of sludge (12-36 g/L) were tested for different values of ES (7000-75,000 kJ/kgTS) and 28 g/L was found as the optimum value according to the solubilized chemical oxygen demand in the liquid phase (SCOD). PUS of 75-150 W was investigated under controlled temperature and the "high power input - short duration" procedure was the most effective at a given ES. The temperature increase in adiabatic US application significantly improved SCOD compared to isothermal conditions. With PUS of 150 W, the effect of external pressure was investigated in the range of 1-16 bar under isothermal and adiabatic conditions for two types of sludge: an optimum pressure of about 2 bar was found regardless of temperature conditions and ES values. Under isothermal conditions, the resulting improvement of sludge disintegration efficacy as compared to atmospheric pressure was by 22-67% and 26-37% for mixed and secondary sludge, respectively. Besides, mean particle diameter (D[4,3]) of the three sludge types decreased respectively from 408, 117, and 110 μm to about 94-97, 37-42, and 36-40 μm regardless of sonication conditions, and the size reduction process was much faster than COD extraction. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Pretreatment techniques for biofuels and biorefineries

    Energy Technology Data Exchange (ETDEWEB)

    Fang, Zhen (ed.) [Chinese Academy of Sciences, Kunming, YN (China). Xishuangbanna Tropical Botonical Garden

    2013-02-01

    The first book focused on pretreatment techniques for biofuels contributed by the world's leading experts. Extensively covers the different types of biomass, various pretreatment approaches and methods that show the subsequent production of biofuels and chemicals. In addition to traditional pretreatment methods, novel techniques are also introduced and discussed. An accessible reference work for students, researchers, academicians and industrialists in biorefineries. This book includes 19 chapters contributed by the world's leading experts on pretreatment methods for biomass. It extensively covers the different types of biomass (e.g. molasses, sugar beet pulp, cheese whey, sugarcane residues, palm waste, vegetable oil, straws, stalks and wood), various pretreatment approaches (e.g. physical, thermal, chemical, physicochemical and biological) and methods that show the subsequent production of biofuels and chemicals such as sugars, ethanol, extracellular polysaccharides, biodiesel, gas and oil. In addition to traditional methods such as steam, hot-water, hydrothermal, diluted-acid, organosolv, ozonolysis, sulfite, milling, fungal and bacterial, microwave, ultrasonic, plasma, torrefaction, pelletization, gasification (including biogas) and liquefaction pretreatments, it also introduces and discusses novel techniques such as nano and solid catalysts, organic electrolyte solutions and ionic liquids. This book offers a review of state-of-the-art research and provides guidance for the future paths of developing pretreatment techniques of biomass for biofuels, especially in the fields of biotechnology, microbiology, chemistry, materials science and engineering. It intends to provide a systematic introduction of pretreatment techniques. It is an accessible reference work for students, researchers, academicians and industrialists in biorefineries.

  17. Broadly reactive human CD8 T cells that recognize an epitope conserved between VZV, HSV and EBV.

    Directory of Open Access Journals (Sweden)

    Christopher Chiu

    2014-03-01

    Full Text Available Human herpesviruses are important causes of potentially severe chronic infections for which T cells are believed to be necessary for control. In order to examine the role of virus-specific CD8 T cells against Varicella Zoster Virus (VZV, we generated a comprehensive panel of potential epitopes predicted in silico and screened for T cell responses in healthy VZV seropositive donors. We identified a dominant HLA-A*0201-restricted epitope in the VZV ribonucleotide reductase subunit 2 and used a tetramer to analyze the phenotype and function of epitope-specific CD8 T cells. Interestingly, CD8 T cells responding to this VZV epitope also recognized homologous epitopes, not only in the other α-herpesviruses, HSV-1 and HSV-2, but also the γ-herpesvirus, EBV. Responses against these epitopes did not depend on previous infection with the originating virus, thus indicating the cross-reactive nature of this T cell population. Between individuals, the cells demonstrated marked phenotypic heterogeneity. This was associated with differences in functional capacity related to increased inhibitory receptor expression (including PD-1 along with decreased expression of co-stimulatory molecules that potentially reflected their stimulation history. Vaccination with the live attenuated Zostavax vaccine did not efficiently stimulate a proliferative response in this epitope-specific population. Thus, we identified a human CD8 T cell epitope that is conserved in four clinically important herpesviruses but that was poorly boosted by the current adult VZV vaccine. We discuss the concept of a "pan-herpesvirus" vaccine that this discovery raises and the hurdles that may need to be overcome in order to achieve this.

  18. Pretreatment Mitochondrial Priming Correlates with Clinical Response to Cytotoxic Chemotherapy

    Science.gov (United States)

    Chonghaile, Triona Ni; Sarosiek, Kristopher A.; Vo, Thanh-Trang; Ryan, Jeremy A.; Tammareddi, Anupama; Moore, Victoria Del Gaizo; Deng, Jing; Anderson, Ken; Richardson, Paul; Tai, Yu-Tzu; Mitsiades, Constantine S.; Matulonis, Ursula A.; Drapkin, Ronny; Stone, Richard; DeAngelo, Daniel J.; McConkey, David J.; Sallan, Stephen E.; Silverman, Lewis; Hirsch, Michelle S.; Carrasco, Daniel Ruben; Letai, Anthony

    2011-01-01

    Cytotoxic chemotherapy targets elements common to all nucleated human cells, such as DNA and microtubules, yet it selectively kills tumor cells. Here we show that clinical response to these drugs correlates with, and may be partially governed by, the pre-treatment proximity of tumor cell mitochondria to the apoptotic threshold, a property called mitochondrial priming. We used BH3 profiling to measure priming in tumor cells from patients with multiple myeloma, acute myelogenous and lymphoblastic leukemia, and ovarian cancer. This assay measures mitochondrial response to peptides derived from pro-apoptotic BH3 domains of proteins critical for death signaling to mitochondria. Patients with highly primed cancers exhibited superior clinical response to chemotherapy. In contrast, chemoresistant cancers and normal tissues were poorly primed. Manipulation of mitochondrial priming might enhance the efficacy of cytotoxic agents. PMID:22033517

  19. Prevalence and risk factors for oral DNA tumor viruses in HIV-infected youth.

    Science.gov (United States)

    Kahn, Jessica A; Rudy, Bret J; Xu, Jiahong; Kapogiannis, Bill; Secord, Elizabeth; Gillison, Maura

    2016-11-01

    Human papillomavirus (HPV), Epstein-Barr virus (EBV), and Kaposi sarcoma-associated herpes virus (KSHV) may promote oral cancers, especially among immunosuppressed individuals. The aims of this study were to examine whether demographic characteristics, medical history, sexual behaviors, substance use, CD4+ T-cell count, HIV viral load, and HPV vaccination were associated with HPV, EBV, and KSHV infection and viral load. Multivariable modeling using logistic or linear regression examined associations between independent variables and infection or viral load, respectively. Among 272 HIV-infected 12-24-year-old youth, 19.5% were positive for oral HPV, 88.2% for EBV, and 11.8% for KSHV. In multivariable models, recent marijuana use (OR 1.97, 95%CI 1.02-3.82) and lower CD4+ T-cell count (HIV viral load (>400 vs. HIV diagnosis (1-14 vs. 18-20 years: EC 0.33, SE 0.16, P = 0.049; 15-17 vs. 18-20 years: EC 0.35, SE 0.13, P = 0.0099) with KSHV viral load. In conclusion, substance use and immunosuppression are associated with oral DNA tumor viruses in HIV-infected youth. J. Med. Virol. 88:1944-1952, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  20. Microwave pretreatment of switchgrass for bioethanol production

    Science.gov (United States)

    Keshwani, Deepak Radhakrishin

    Lignocellulosic materials are promising alternative feedstocks for bioethanol production. These materials include agricultural residues, cellulosic waste such as newsprint and office paper, logging residues, and herbaceous and woody crops. However, the recalcitrant nature of lignocellulosic biomass necessitates a pretreatment step to improve the yield of fermentable sugars. The overall goal of this dissertation is to expand the current state of knowledge on microwave-based pretreatment of lignocellulosic biomass. Existing research on bioenergy and value-added applications of switchgrass is reviewed in Chapter 2. Switchgrass is an herbaceous energy crop native to North America and has high biomass productivity, potentially low requirements for agricultural inputs and positive environmental impacts. Based on results from test plots, yields in excess of 20 Mg/ha have been reported. Environmental benefits associated with switchgrass include the potential for carbon sequestration, nutrient recovery from run-off, soil remediation and provision of habitats for grassland birds. Published research on pretreatment of switchgrass reported glucose yields ranging from 70-90% and xylose yields ranging from 70-100% after hydrolysis and ethanol yields ranging from 72-92% after fermentation. Other potential value-added uses of switchgrass include gasification, bio-oil production, newsprint production and fiber reinforcement in thermoplastic composites. Research on microwave-based pretreatment of switchgrass and coastal bermudagrass is presented in Chapter 3. Pretreatments were carried out by immersing the biomass in dilute chemical reagents and exposing the slurry to microwave radiation at 250 watts for residence times ranging from 5 to 20 minutes. Preliminary experiments identified alkalis as suitable chemical reagents for microwave-based pretreatment. An evaluation of different alkalis identified sodium hydroxide as the most effective alkali reagent. Under optimum pretreatment

  1. Enzymatic hydrolysis of pretreated soybean straw

    International Nuclear Information System (INIS)

    Xu Zhong; Wang Qunhui; Jiang Zhaohua; Yang Xuexin; Ji Yongzhen

    2007-01-01

    In order to produce lactic acid, from agricultural residues such as soybean straw, which is a raw material for biodegradable plastic production, it is necessary to decompose the soybean straw into soluble sugars. Enzymatic hydrolysis is one of the methods in common use, while pretreatment is the effective way to increase the hydrolysis rate. The optimal conditions of pretreatment using ammonia and enzymatic hydrolysis of soybean straw were determined. Compared with the untreated straw, cellulose in straw pretreated by ammonia liquor (10%) soaking for 24 h at room temperature increased 70.27%, whereas hemicellulose and lignin in pretreated straw decreased to 41.45% and 30.16%, respectively. The results of infrared spectra (IR), scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis also showed that the structure and the surface of the straw were changed through pretreatment that is in favor of the following enzymatic hydrolysis. maximum enzymatic hydrolysis rate of 51.22% was achieved at a substrate concentration of 5% (w/v) at 50 deg. C and pH 4.8 using cellulase (50 fpu/g of substrate) for 36 h

  2. Temperature Modelling of the Biomass Pretreatment Process

    DEFF Research Database (Denmark)

    Prunescu, Remus Mihail; Blanke, Mogens; Jensen, Jakob M.

    2012-01-01

    In a second generation biorefinery, the biomass pretreatment stage has an important contribution to the efficiency of the downstream processing units involved in biofuel production. Most of the pretreatment process occurs in a large pressurized thermal reactor that presents an irregular temperature...... distribution. Therefore, an accurate temperature model is critical for observing the biomass pretreatment. More than that, the biomass is also pushed with a constant horizontal speed along the reactor in order to ensure a continuous throughput. The goal of this paper is to derive a temperature model...... that captures the environmental temperature differences inside the reactor using distributed parameters. A Kalman filter is then added to account for any missing dynamics and the overall model is embedded into a temperature soft sensor. The operator of the plant will be able to observe the temperature in any...

  3. Optimizing hyaluronidase dose and plasmid DNA delivery greatly improves gene electrotransfer efficiency in rat skeletal muscle

    DEFF Research Database (Denmark)

    Åkerström, Thorbjörn; Vedel, Kenneth; Needham Andersen, Josefine

    2015-01-01

    Transfection of rat skeletal muscle in vivo is a widely used research model. However, gene electrotransfer protocols have been developed for mice and yield variable results in rats. We investigated whether changes in hyaluronidase pre-treatment and plasmid DNA delivery can improve transfection...... efficiency in rat skeletal muscle. We found that pre-treating the muscle with a hyaluronidase dose suitable for rats (0.56. U/g b.w.) prior to plasmid DNA injection increased transfection efficiency by >200% whereas timing of the pre-treatment did not affect efficiency. Uniformly distributing plasmid DNA...... with a homogenous distribution. We also show that transfection was stable over five weeks of regular exercise or inactivity. Our findings show that species-specific plasmid DNA delivery and hyaluronidase pre-treatment greatly improves transfection efficiency in rat skeletal muscle....

  4. Cleanser, Detergent, Personal Care Product Pretreatment Evaluation

    Science.gov (United States)

    Adam, Niklas

    2010-01-01

    The purpose of the Cleanser, Detergent, Personal Care Product, and Pretreatment Evaluation & Selection task is to identify the optimal combination of personal hygiene products, crew activities, and pretreatment strategies to provide the crew with sustainable life support practices and a comfortable habitat. Minimal energy, mass, and crew time inputs are desired to recycle wastewater during long duration missions. This document will provide a brief background on the work this past year supporting the ELS Distillation Comparison Test, issues regarding use of the hygiene products originally chosen for the test, methods and results used to select alternative products, and lessons learned from testing.

  5. Mitochondrial DNA.

    Science.gov (United States)

    Wright, Russell G.; Bottino, Paul J.

    1986-01-01

    Provides background information for teachers on mitochondrial DNA, pointing out that it may have once been a free-living organism. Includes a ready-to-duplicate exercise titled "Using Microchondrial DNA to Measure Evolutionary Distance." (JN)

  6. Modeling DNA

    Science.gov (United States)

    Robertson, Carol

    2016-01-01

    Deoxyribonucleic acid (DNA) is life's most amazing molecule. It carries the genetic instructions that almost every organism needs to develop and reproduce. In the human genome alone, there are some three billion DNA base pairs. The most difficult part of teaching DNA structure, however, may be getting students to visualize something as small as a…

  7. Fermentation of pretreated corncob hemicellulose hydrolysate to ...

    African Journals Online (AJOL)

    To investigate the effect of unknown fermentation inhibitors in corncob hemicellulose acid hydrolysate processed by pretreatment and detoxification on fermentation, corncob hemicellulose acid hydrolysate and artificially prepared hydrolysate were fermented in parallel by Candida shehatae YHFK-2. The results show that ...

  8. Antibacterial effect of surface pretreatment techniques against ...

    African Journals Online (AJOL)

    (CFU/ml), respectively, among the groups evaluated (P < 0.05). Er:YAG laser irradiation and its combinations with other antibacterial surface pretreatment applications also inhibited the bacterial growth with, respectively, 1444, 406, and 294 CFU/ml bacterial recovery being more efficient than KTP laser irradiation and ozone ...

  9. Antibacterial Effect of Surface Pretreatment Techniques against ...

    African Journals Online (AJOL)

    2018-02-23

    Feb 23, 2018 ... of dentin chips were obtained from the cavity walls, and the number of bacteria recovered was counted. Kruskal–Wallis ... Keywords: Antibacterial effect, cavity surface pretreatment techniques, cavity preparation, dental, dental ... wavelengths for removing oral soft and dental hard tissues without pain relief, ...

  10. Bromine pretreated chitosan for adsorption of lead

    Indian Academy of Sciences (India)

    Pollution by heavy metals like lead (II) is responsible for health hazards and environmental degradation. Adsorption is a prevalent method applied for removal of heavy metal pollutants from water. This study explored adsorption performances of 30% bromine pretreated chitosan for lead (II) abatement from water. Bromine ...

  11. The Safety and Immunological Effects of rAd5-EBV-LMP2 Vaccine in Nasopharyngeal Carcinoma Patients: A Phase I Clinical Trial and Two-Year Follow-Up.

    Science.gov (United States)

    Si, Yongfeng; Deng, Zhuoxia; Lan, Guiping; Du, Haijun; Wang, Yongli; Si, Jinyuan; Wei, Jiazhang; Weng, Jingjin; Qin, Yangda; Huang, Bo; Yang, Yong; Qin, Ying

    2016-01-01

    Epstein-Barr virus (EBV)-encoded latent membrane protein 2 (LMP2) promotes nasopharyngeal carcinoma (NPC) progression. Previously, we reported that the dendritic cells (DCs) transfected with EBV-LMP2 recombinant serotype 5 adenoviruses (rAd5) induced anti-tumor effect by eliciting cytotoxic T lymphocytes (CTLs)-mediated immune response in vitro and the adenoviral vaccine of EBV-LMP2 (rAd5-EBV-LMP2) stimulated antigen-specific cellular immunity in mice. However, the safety and immunological effect of rAd5-EBV-LMP2 vaccine in human still remained unknown. Here we conducted a single-center, non-randomized, open-label, single-arm phase I clinical trial to clarify this unsolved issue. A total of 24 patients with regional advanced NPC were sequentially enrolled into three dose level groups (2×10(9), 2×10(10), 2×10(11) vp). The rAd5-EBV-LMP2 vaccines were intramuscularly injected for four times within 28 d (D0, D7, D14, D28). Blood samples were harvested immediately before every vaccination, one week and one month after the last vaccination (D0, D7, D14, D28, D35, D58). All the vaccine inoculation-related toxicities presented as grade I/II adverse events. The most frequent systemic adverse reactions were fatigue (33.0%, 8/24), myalgia (29.2%, 7/24) and cough (29.2%, 7/24), while the most common regional adverse reaction was tenderness in the inoculation site (54.2%, 13/24). In addition, proportion of CD(3+)CD(4+) cells in peripheral blood was significantly increased in the high dose group (2×10(11) vp). The rAd5-EBV-LMP2 vaccine was generally well-tolerated and the high dose (2×10(11) vp) is recommended to be adopted in phase II studies. The long-term outcome of rAd5-EBV-LMP2 vaccine inoculation is required to be determined in following placebo-controlled trials.

  12. Dna Sequencing

    Science.gov (United States)

    Tabor, Stanley; Richardson, Charles C.

    1995-04-25

    A method for sequencing a strand of DNA, including the steps off: providing the strand of DNA; annealing the strand with a primer able to hybridize to the strand to give an annealed mixture; incubating the mixture with four deoxyribonucleoside triphosphates, a DNA polymerase, and at least three deoxyribonucleoside triphosphates in different amounts, under conditions in favoring primer extension to form nucleic acid fragments complementory to the DNA to be sequenced; labelling the nucleic and fragments; separating them and determining the position of the deoxyribonucleoside triphosphates by differences in the intensity of the labels, thereby to determine the DNA sequence.

  13. DNA Damage in Inflammation-Related Carcinogenesis and Cancer Stem Cells

    Directory of Open Access Journals (Sweden)

    Shiho Ohnishi

    2013-01-01

    Full Text Available Infection and chronic inflammation have been recognized as important factors for carcinogenesis. Under inflammatory conditions, reactive oxygen species (ROS and reactive nitrogen species (RNS are generated from inflammatory and epithelial cells and result in oxidative and nitrative DNA damage, such as 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG and 8-nitroguanine. The DNA damage can cause mutations and has been implicated in the initiation and/or promotion of inflammation-mediated carcinogenesis. It has been estimated that various infectious agents are carcinogenic to humans (IARC group 1, including parasites (Schistosoma haematobium (SH and Opisthorchis viverrini (OV, viruses (hepatitis C virus (HCV, human papillomavirus (HPV, and Epstein-Barr virus (EBV, and bacterium Helicobacter pylori (HP. SH, OV, HCV, HPV, EBV, and HP are important risk factors for bladder cancer, cholangiocarcinoma, hepatocellular carcinoma, cervical cancer, nasopharyngeal carcinoma, and gastric cancer, respectively. We demonstrated that 8-nitroguanine was strongly formed via inducible nitric oxide synthase (iNOS expression at these cancer sites of patients. Moreover, 8-nitroguanine was formed in Oct3/4-positive stem cells in SH-associated bladder cancer tissues and in Oct3/4- and CD133-positive stem cells in OV-associated cholangiocarcinoma tissues. Therefore, it is considered that oxidative and nitrative DNA damage in stem cells may play a key role in inflammation-related carcinogenesis.

  14. Investigação da LMP1 do EBV e a coinfeçcão do HPV em lesões genitais de pacientes infectados ou não pelo HIV Investigation of the LMP1 EBV and co-infection by HPV in genital lesions of patients infected or not by HIV

    Directory of Open Access Journals (Sweden)

    Fabiana Resende Rodrigues

    2010-10-01

    Full Text Available INTRODUÇÃO: Vários estudos têm demonstrado associação do vírus Epstein-Barr (EBV com neoplasias malignas, inclusive genitais, em que o papilomavírus humano (HPV é o principal vírus associado às neoplasias epiteliais benignas e malignas. OBJETIVO: Investigar a presença do EBV e do HPV em lesões genitais de ambos os sexos, em pacientes soropositivos (grupo A ou não (grupo B para o vírus da imunodeficiência humana (HIV. MATERIAL E MÉTODO: Selecionados 126 pacientes e 135 lesões anogenitais, sendo 67 pacientes (53% e 75 lesões (56% no grupo A e 59 pacientes (47% e 60 lesões (44% no grupo B, para análise imuno-histoquímica (IHQ por meio dos anticorpos monoclonais antiproteína latente de membrana 1 (LMP1 e HPV (DAKO®. RESULTADOS: A análise mostrou que o número total de lesões com imunopositividade para o HPV e para a LMP1 foi maior no grupo A (32 e 35, respectivamente quando comparado ao B (16 e seis, respectivamente. A análise estatística (nível de significância de 5% mostrou que as proporções para o HPV não são estatisticamente significativas (z = 1,93; valor p = 0,053. Entretanto, para a LMP1, a diferença (47% no grupo A e 10% no B é significativa (z = 4,60; valor p = 4,2×10-6. Do mesmo modo, a associação HPV-LMP1 (21% no grupo A e 7% no B também mostrou diferença estatisticamente significativa (z = 2,38; valor p = 0,017. CONCLUSÃO: Esses resultados indicam a possibilidade de sinergismo da infecção pelo EBV e a coinfecção EBV-HPV em lesões epiteliais genitais, particularmente em pacientes soropositivos para o HIV. Entretanto, investigações com metodologia de maior especificidade e sensibilidade são necessárias para a verificação da real participação do EBV na patogênese de lesões epiteliais genitais.INTRODUCTION: Several studies have demonstrated the association between Epstein-Barr virus (EBV and malignant neoplasias, including genital lesions, in which the human papillomavirus (HPV is the

  15. Epstein-Barr Virus Rta-Mediated Accumulation of DNA Methylation Interferes with CTCF Binding in both Host and Viral Genomes.

    Science.gov (United States)

    Chen, Yen-Ju; Chen, Yu-Lian; Chang, Yao; Wu, Chung-Chun; Ko, Ying-Chieh; Tsao, Sai Wah; Chen, Jen-Yang; Lin, Su-Fang

    2017-08-01

    Rta, an Epstein-Barr virus (EBV) immediate-early protein, reactivates viral lytic replication that is closely associated with tumorigenesis. In previous studies, we demonstrated that in epithelial cells Rta efficiently induced cellular senescence, which is an irreversible G 1 arrest likely to provide a favorable environment for productive replications of EBV and Kaposi's sarcoma-associated herpesvirus (KSHV). To restrict progression of the cell cycle, Rta simultaneously upregulates CDK inhibitors and downregulates MYC, CCND1, and JUN, among others. Rta has long been known as a potent transcriptional activator, thus its role in gene repression is unexpected. In silico analysis revealed that the promoter regions of MYC , CCND1 , and JUN are common in (i) the presence of CpG islands, (ii) strong chromatin immunoprecipitation (ChIP) signals of CCCTC-binding factor (CTCF), and (iii) having at least one Rta binding site. By combining ChIP assays and DNA methylation analysis, here we provide evidence showing that Rta binding accumulated CpG methylation and decreased CTCF occupancy in the regulatory regions of MYC , CCND1 , and JUN , which were associated with downregulated gene expression. Stable residence of CTCF in the viral latency and reactivation control regions is a hallmark of viral latency. Here, we observed that Rta-mediated decreased binding of CTCF in the viral genome is concurrent with virus reactivation. Via interfering with CTCF binding, in the host genome Rta can function as a transcriptional repressor for gene silencing, while in the viral genome Rta acts as an activator for lytic gene loci by removing a topological constraint established by CTCF. IMPORTANCE CTCF is a multifunctional protein that variously participates in gene expression and higher-order chromatin structure of the cellular and viral genomes. In certain loci of the genome, CTCF occupancy and DNA methylation are mutually exclusive. Here, we demonstrate that the Epstein-Barr virus (EBV

  16. Pretreatment of Latent Prints for Laser Development.

    Science.gov (United States)

    Menzel, E R

    1989-06-01

    The pretreatment procedures for laser detection of latent fingerprints is reviewed. The general features of laser detection and the operational aspects of the examination of physical evidence for laser fingerprint detection are enumerated in the initial sections. The literature review is divided into various pretreatment approaches. Cited studies prior to 1981 are primarily concerned with the demonstration of the viability of laser fingerprint detection, whereas work post-1981 addresses issues of compatibility with the traditional methods of fingerprint development, examination of difficult surfaces, and the routine implementation of laser detection by law enforcement agencies. Related topics of research, review articles, conference reports, case examinations, the research support climate, and future trends are also briefly addressed. Copyright © 1989 Central Police University.

  17. Pretreatment of Dioscorea zingiberensis for Microbial Transformation

    Directory of Open Access Journals (Sweden)

    Tianxiang Zheng

    2014-09-01

    Full Text Available The influences of five pretreatments on fungal growth and enzyme production during microbial transformation of Dioscorea zingiberensis (DZW were studied. The biomass, α-rhamnase and β-glucosidase activities in the fermentation system were employed in the study to determine how each method affected the efficiency of microbial transformation. The fungal strain grew better on the substrate which contained easily utilized carbon source. While lack of carbon source induced the strain produce more glucosidase. Among five pretreatment methods, complex enzymatic hydrolyzation can remove 84.3% starch and 76.5% fibre from DZW in form of sugar, which resulted in high α-rhamnase activity of 2.89 IU/mL and β-glucosidase activity of 8.17 IU/mL in fermentation broth.

  18. Influence of amino acids Shiff bases on irradiated DNA stability in vivo.

    Science.gov (United States)

    Karapetyan, N H; Malakyan, M H; Bajinyan, S A; Torosyan, A L; Grigoryan, I E; Haroutiunian, S G

    2013-01-01

    To reveal protective role of the new Mn(II) complexes with Nicotinyl-L-Tyrosinate and Nicotinyl-L-Tryptophanate Schiff Bases against ionizing radiation. The DNA of the rats liver was isolated on 7, 14, and 30 days after X-ray irradiation. The differences between the DNA of irradiated rats and rats pre-treated with Mn(II) complexes were studied using the melting, microcalorimetry, and electrophoresis methods. The melting parameters and the melting enthalpy of rats livers DNA were changed after the X-ray irradiation: melting temperature and melting enthalpy were decreased and melting interval was increased. These results can be explained by destruction of DNA molecules. It was shown that pre-treatment of rats with Mn(II) complexes approximates the melting parameters to norm. Agarose gel electrophoresis data confirmed the results of melting studies. The separate DNA fragments were revealed in DNA samples isolated from irradiated animals. The DNA isolated from animals pre-treated with the Mn(II) chelates had better electrophoretic characteristics, which correspond to healthy DNA. Pre-treatment of the irradiated rats with Mn(II)(Nicotinil-L-Tyrosinate) and Mn(II)(Nicotinil-L-Tryptophanate)2 improves the DNA characteristics.

  19. Pretreatment Capabilities and Benefits of Electrocoagulation

    Science.gov (United States)

    2004-12-01

    4.1.1.2 Oil-Water Emulsions – Mock Solutions ………………… 20 4.1.1.3 Tannin and Humic Acid Mixture – Mock Solutions …….. 35 4.1.1.4 Silt – Mock...water Influent/ effluent water control Industrial wastewater Pre-treatment for drinking water Oil emulsion breaking Medical waste

  20. Hydrothermal pretreatments of macroalgal biomass for biorefineries

    DEFF Research Database (Denmark)

    Ruiz, Héctor A.; Rodríguez-Jasso, Rosa M.; Aguedo, Mario

    2015-01-01

    ecently, macroalgal biomass is gaining wide attention as an alternative in the production of biofuels (as bioetanol and biogas) and compounds with high added value with specific properties (antioxidants, anticoagulants, anti–inflammatories) for applications in food, medical and energy industries...... in accordance with the integrated biorefineries. Furthermore, biorefinery concept requires processes that allow efficient utilization of all components of the biomass. The pretreatment step in a biorefinery is often based on hydrothermal principles of high temperatures in aqueous solution. Therefore...

  1. Waste Separations and Pretreatment Workshop report

    International Nuclear Information System (INIS)

    Cruse, J.M.; Harrington, R.A.; Quadrel, M.J.

    1994-01-01

    This document provides the minutes from the Waste Separations and Pretreatment Workshop sponsored by the Underground Storage Tank-Integrated Demonstration in Salt Lake City, Utah, February 3--5, 1993. The Efficient Separations and Processing-Integrated Program and the Hanford Site Tank Waste Remediation System were joint participants. This document provides the detailed minutes, including responses to questions asked, an attendance list, reproductions of the workshop presentations, and a revised chart showing technology development activities

  2. Phosphoproteomic Profiling Reveals Epstein-Barr Virus Protein Kinase Integration of DNA Damage Response and Mitotic Signaling.

    Directory of Open Access Journals (Sweden)

    Renfeng Li

    2015-12-01

    Full Text Available Epstein-Barr virus (EBV is etiologically linked to infectious mononucleosis and several human cancers. EBV encodes a conserved protein kinase BGLF4 that plays a key role in the viral life cycle. To provide new insight into the host proteins regulated by BGLF4, we utilized stable isotope labeling by amino acids in cell culture (SILAC-based quantitative proteomics to compare site-specific phosphorylation in BGLF4-expressing Akata B cells. Our analysis revealed BGLF4-mediated hyperphosphorylation of 3,046 unique sites corresponding to 1,328 proteins. Frequency analysis of these phosphosites revealed a proline-rich motif signature downstream of BGLF4, indicating a broader substrate recognition for BGLF4 than its cellular ortholog cyclin-dependent kinase 1 (CDK1. Further, motif analysis of the hyperphosphorylated sites revealed enrichment in ATM, ATR and Aurora kinase substrates while functional analyses revealed significant enrichment of pathways related to the DNA damage response (DDR, mitosis and cell cycle. Phosphorylation of proteins associated with the mitotic spindle assembly checkpoint (SAC indicated checkpoint activation, an event that inactivates the anaphase promoting complex/cyclosome, APC/C. Furthermore, we demonstrated that BGLF4 binds to and directly phosphorylates the key cellular proteins PP1, MPS1 and CDC20 that lie upstream of SAC activation and APC/C inhibition. Consistent with APC/C inactivation, we found that BGLF4 stabilizes the expression of many known APC/C substrates. We also noted hyperphosphorylation of 22 proteins associated the nuclear pore complex, which may contribute to nuclear pore disassembly and SAC activation. A drug that inhibits mitotic checkpoint activation also suppressed the accumulation of extracellular EBV virus. Taken together, our data reveal that, in addition to the DDR, manipulation of mitotic kinase signaling and SAC activation are mechanisms associated with lytic EBV replication. All MS data have been

  3. Lack of dependence on p53 for DNA double strand break repair of episomal vectors in human lymphoblasts

    Science.gov (United States)

    Kohli, M.; Jorgensen, T. J.

    1999-01-01

    The p53 tumor suppressor gene has been shown to be involved in a variety of repair processes, and recent findings have suggested that p53 may be involved in DNA double strand break repair in irradiated cells. The role of p53 in DNA double strand break repair, however, has not been fully investigated. In this study, we have constructed a novel Epstein-Barr virus (EBV)-based shuttle vector, designated as pZEBNA, to explore the influence of p53 on DNA strand break repair in human lymphoblasts, since EBV-based vectors do not inactivate the p53 pathway. We have compared plasmid survival of irradiated, restriction enzyme linearized, and calf intestinal alkaline phosphatase (CIP)-treated pZEBNA with a Simian virus 40 (SV40)-based shuttle vector, pZ189, in TK6 (wild-type p53) and WTK1 (mutant p53) lymphoblasts and determined that p53 does not modulate DNA double strand break repair in these cell lines. Copyright 1999 Academic Press.

  4. Cost comparison for REDC pretreatment project

    International Nuclear Information System (INIS)

    Robinson, S.M.; Homan, F.J.

    1997-06-01

    This analysis has been prepared to support the planned expenditure to provide the Radiochemical Engineering Development Center (REDC) with the capability to pretreat their liquid low-level waste (LLLW) before discharging it to the Oak Ridge National Laboratory (ORNL) LLLW system. Pretreatment will remove most of the radioactivity, particularly the transuranic isotopes and Cs-137 from the waste to be discharged. This will render the supernates that accumulate in the storage tanks low-activity Class B low-level wastes rather than high-activity Class B or Class C wastes. The sludges will be Class C rather than remote-handled transuranic (RH-TRU) wastes. When REDC wastes are commingled with other ORNL LLLW, the present-worth treatment and transport costs are higher by a factor of 1.3 for the no-pretreatment cases. This result is consistent with data from similar studies conducted at other sites. Based on the information presented in this analysis, the recommendation is to proceed with REDC treatment projects

  5. Pretreatment of Agave americana stalk for enzymatic saccharification.

    Science.gov (United States)

    Yang, Qiang; Pan, Xuejun

    2012-12-01

    Agave americana is one of commonly grown agave species but currently less valuable because its large flower stalk cannot be used for producing alcoholic beverage. In the present study, the stalk was pretreated with dilute acid (DA), sulfite (SPORL), and sodium hydroxide (NaOH) to preliminarily assess its potential as feedstock for bioethanol production. The changes of cell wall components during the pretreatments, enzymatic digestibility of the pretreated stalks, and the adsorption of cellulases on the substrates were investigated. Results indicated that the pretreatments significantly improved the enzymatic digestibility of the agave stalk. SPORL pretreatment gave higher substrate and sugar yields, while NaOH pretreated stalk had better digestibility under the investigated conditions. The better hydrolysability of NaOH-pretreated stalk was attributed to low lignin and hemicellulose content and high affinity to cellulases. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Oncogenic Viruses and Breast Cancer: Mouse Mammary Tumor Virus (MMTV, Bovine Leukemia Virus (BLV, Human Papilloma Virus (HPV, and Epstein–Barr Virus (EBV

    Directory of Open Access Journals (Sweden)

    James S. Lawson

    2018-01-01

    Full Text Available BackgroundAlthough the risk factors for breast cancer are well established, namely female gender, early menarche and late menopause plus the protective influence of early pregnancy, the underlying causes of breast cancer remain unknown. The development of substantial recent evidence indicates that a handful of viruses may have a role in breast cancer. These viruses are mouse mammary tumor virus (MMTV, bovine leukemia virus (BLV, human papilloma viruses (HPVs, and Epstein–Barr virus (EBV-also known as human herpes virus type 4. Each of these viruses has documented oncogenic potential. The aim of this review is to inform the scientific and general community about this recent evidence.The evidenceMMTV and human breast cancer—the evidence is detailed and comprehensive but cannot be regarded as conclusive. BLV and human breast cancer—the evidence is limited. However, in view of the emerging information about BLV in human breast cancer, it is prudent to encourage the elimination of BLV in cattle, particularly in the dairy industry. HPVs and breast cancer—the evidence is substantial but not conclusive. The availability of effective preventive vaccines is a major advantage and their use should be encouraged. EBV and breast cancer—the evidence is also substantial but not conclusive. Currently, there are no practical means of either prevention or treatment. Although there is evidence of genetic predisposition, and cancer in general is a culmination of events, there is no evidence that inherited genetic traits are causal.ConclusionThe influence of oncogenic viruses is currently the major plausible hypothesis for a direct cause of human breast cancer.

  7. Soro-epidemiologia do virus de Epstein-Barr (VEB The seroepidemiology of the Epstein-Barr vírus (EBV

    Directory of Open Access Journals (Sweden)

    Claudio Sergio Pannuti

    1981-02-01

    Full Text Available É feita revisão dos estudos soro-epidemiológicos do vírus de Epstein-Barr (vírus EB, mostrando a ubicuidade deste vírus, bem como sua relação com doenças malígnas e não malígnas do homem. O desenvolvimento de novas técnicas laboratoriais e o acúmulo, nos últimos 10 anos, de grande número de estudos soro-epidemiológicos, levaram à conclusão de que, além da associação etiológica com a mononucleose infecciosa, torna-se cada vez mais evidente a ligação do vírus EB com o linfoma de Burkitt e com o carcinoma do nasofaringe. Estas observações, além de sua extraordinária importância intrínseca, tem sido de grande utilidade para a melhor compreensão dos possíveis mecanismos e efeitos das infecções virais no homem.Since the discovery of the Epstein-Barr virus (EBV in 1964, seroepidemiological studies have sought to characterize this member of the Herpetoviridae family. Through these studies the ubiquity of EBV became evident, as did its relationship to human malignant and non-malignant diseases. Besides being linked etiologically with infectious mononucleosis, the Epstein-Barr virus is closely associated with Burkitt lymphoma and nasopharingeal carcinoma. In addition to the intrinsic value of the findings reported, these extremely important studies have also brought a better understanding of the possible mechanisms and affects of viral infections in man.

  8. Oncogenic Viruses and Breast Cancer: Mouse Mammary Tumor Virus (MMTV), Bovine Leukemia Virus (BLV), Human Papilloma Virus (HPV), and Epstein-Barr Virus (EBV).

    Science.gov (United States)

    Lawson, James S; Salmons, Brian; Glenn, Wendy K

    2018-01-01

    Although the risk factors for breast cancer are well established, namely female gender, early menarche and late menopause plus the protective influence of early pregnancy, the underlying causes of breast cancer remain unknown. The development of substantial recent evidence indicates that a handful of viruses may have a role in breast cancer. These viruses are mouse mammary tumor virus (MMTV), bovine leukemia virus (BLV), human papilloma viruses (HPVs), and Epstein-Barr virus (EBV-also known as human herpes virus type 4). Each of these viruses has documented oncogenic potential. The aim of this review is to inform the scientific and general community about this recent evidence. MMTV and human breast cancer-the evidence is detailed and comprehensive but cannot be regarded as conclusive. BLV and human breast cancer-the evidence is limited. However, in view of the emerging information about BLV in human breast cancer, it is prudent to encourage the elimination of BLV in cattle, particularly in the dairy industry. HPVs and breast cancer-the evidence is substantial but not conclusive. The availability of effective preventive vaccines is a major advantage and their use should be encouraged. EBV and breast cancer-the evidence is also substantial but not conclusive. Currently, there are no practical means of either prevention or treatment. Although there is evidence of genetic predisposition, and cancer in general is a culmination of events, there is no evidence that inherited genetic traits are causal. The influence of oncogenic viruses is currently the major plausible hypothesis for a direct cause of human breast cancer.

  9. Possible involvement of ROS generation in vorinostat pretreatment induced enhancement of the antibacterial activity of ciprofloxacin.

    Science.gov (United States)

    Masadeh, Majed M; Alzoubi, Karem H; Al-Azzam, Sayer I; Al-Buhairan, Ahlam M

    2017-01-01

    The mechanism underlying ciprofloxacin action involves interference with transcription and replication of bacterial DNA and, thus, the induction of double-strand breaks in DNA. It also involves elevated oxidative stress, which might contribute to bacterial cell death. Vorinostat was shown to induce oxidative DNA damage. The current work investigated a possible interactive effect of vorinostat on ciprofloxacin-induced cytotoxicity against a number of reference bacteria. Standard bacterial strains were Escherichia coli ATCC 35218, Staphylococcus aureus ATCC29213, Pseudomonas aeruginosa ATCC 9027, Staphylococcus epidermidis ATCC 12228, Acinetobacter baumannii ATCC 17978, Proteus mirabilis ATCC 12459, Klebsiella pneumoniae ATCC 13883, methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 43300), and Streptococcus pneumoniae (ATCC 25923). The antibacterial activity of ciprofloxacin, with or without pretreatment of bacterial cells by vorinostat, was examined using the disc diffusion procedure and determination of the minimum inhibitory concentration (MIC) and zones of inhibition of bacterial growth. All tested bacterial strains showed sensitivity to ciprofloxacin. When pretreated with vorinostat, significantly larger zones of inhibition and smaller MIC values were observed in all bacterial strains compared to those treated with ciprofloxacin alone. In correlation, generation of reactive oxygen species (ROS) induced by the antibacterial action of ciprofloxacin was enhanced by treatment of bacterial cells with vorinostat. Results showed the possible agonistic properties of vorinostat when used together with ciprofloxacin. This could be related to the ability of these agents to enhance oxidative stress in bacterial cells.

  10. DNA glue

    DEFF Research Database (Denmark)

    Filichev, Vyacheslav V; Astakhova, Irina V.; Malakhov, Andrei D.

    2008-01-01

    Significant alterations in thermal stability of parallel DNA triplexes and antiparallel duplexes were observed upon changing the attachment of ethynylpyrenes from para to ortho in the structure of phenylmethylglycerol inserted as a bulge into DNA (TINA). Insertions of two ortho-TINAs as a pseudo...

  11. DNA Vaccines

    Indian Academy of Sciences (India)

    DNA vaccine, immune response, antibodies, infectious diseases. GENERAL I ARTICLE. DNA Vaccines. P N Rangarajan. History of Vaccine Development. The year 1996 marked the 200th anniversary of the first vaccine developed against smallpox by Edward Jenner. In the now- famous 1796 experiment, Jenner scratched ...

  12. Hyperstretching DNA

    NARCIS (Netherlands)

    Schakenraad, Koen; Biebricher, Andreas S.; Sebregts, Maarten; Ten Bensel, Brian; Peterman, Erwin J.G.; Wuite, Gijs J L; Heller, Iddo; Storm, Cornelis; Van Der Schoot, Paul

    2017-01-01

    The three-dimensional structure of DNA is highly susceptible to changes by mechanical and biochemical cues in vivo and in vitro. In particular, large increases in base pair spacing compared to regular B-DNA are effected by mechanical (over)stretching and by intercalation of compounds that are widely

  13. Pretreatment of Reed by Wet Oxidation and Subsequent Utilization of the Pretreated Fibers for Ethanol Production

    DEFF Research Database (Denmark)

    Szijarto, Nora; Kádár, Zsófia; Varga, Eniko

    2009-01-01

    lignocelluloses usually do. In the present study, wet oxidation was investigated as the pretreatment method to enhance the enzymatic digestibility of reed cellulose to soluble sugars and thus improve the convertibility of reed to ethanol. The most effective treatment increased the digestibility of reed cellulose...... by cellulases more than three times compared to the untreated control. During this wet oxidation, 51.7% of the hemicellulose and 58.3% of the lignin were solubilized, whereas 87.1% of the cellulose remained in the solids. After enzymatic hydrolysis of pretreated fibers from the same treatment, the conversion...... of cellulose to glucose was 82.4%. Simultaneous saccharification and fermentation of pretreated solids resulted in a final ethanol concentration as high as 8.7 g/L, yielding 73% of the theoretical....

  14. Unsuitability of lymphoblastoid cell lines as surrogate of cryopreserved isolated lymphocytes for the analysis of DNA double-strand break repair activity

    Energy Technology Data Exchange (ETDEWEB)

    Zijno, Andrea [Department of Environment and Primary Prevention, Istituto Superiore di Sanita, Rome (Italy); Porcedda, Paola [Department of Clinical and Biological Sciences, University of Turin (Italy); Saini, Francesca [Department of Environment and Primary Prevention, Istituto Superiore di Sanita, Rome (Italy); Allione, Alessandra [Institute for Scientific Interchange (ISI) Foundation, Villa Gualino, Turin (Italy); Garofalo, Bruno; Marcon, Francesca [Department of Environment and Primary Prevention, Istituto Superiore di Sanita, Rome (Italy); Guarrera, Simonetta [Institute for Scientific Interchange (ISI) Foundation, Villa Gualino, Turin (Italy); Turinetto, Valentina; Minieri, Valentina [Department of Clinical and Biological Sciences, University of Turin (Italy); Funaro, Ada [Department of Genetics, Biology and Biochemistry, University of Turin (Italy); Crebelli, Riccardo [Department of Environment and Primary Prevention, Istituto Superiore di Sanita, Rome (Italy); Giachino, Claudia [Department of Clinical and Biological Sciences, University of Turin (Italy); Matullo, Giuseppe, E-mail: giuseppe.matullo@unito.it [Institute for Scientific Interchange (ISI) Foundation, Villa Gualino, Turin (Italy); Department of Genetics, Biology and Biochemistry, University of Turin (Italy)

    2010-02-03

    As first task of a comprehensive investigation on DNA repair genotype-phenotype correlations, the suitability of Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCLs) as surrogate of cryopreserved peripheral blood mononuclear cells (PBMCs) in DNA repair phenotypic assays was evaluated. To this aim the amount of DNA damage induced by {gamma}-rays and DNA repair capacity were evaluated in unstimulated (G{sub 0}) and mitogen-simulated (G{sub 2}) PBMC from 20 healthy subjects and in EBV-transformed LCL obtained from the same individuals. Phosphorylation of histone H2AX, micronuclei and chromosomal aberrations were the end-points investigated. The results obtained show higher basal frequencies of binucleated cells bearing micronuclei and nucleoplasmic bridge (NPB) in LCL with respect to PBMC, suggesting that EBV transformation may be associated with chromosomal instability. After irradiation, higher levels of micronuclei were induced in G{sub 0}-treated PBMC compared to cycling LCL; conversely, NPB were more frequent in LCL than in PBMC. Moreover, higher levels of chromosomal aberrations were observed in G{sub 2}-treated PBMC compared to LCL. Concerning {gamma}-H2AX measurements, phosphorylation levels 1 h after treatment and dephosphorylation kinetics were basically similar in LCL and in PBMC. However, while Spearman's test showed a strong correlation between the results obtained in replicated experiments with PBMC, high inter-experimental variability and poor reproducibility was observed in the experiments performed with LCL, possibly due to the intrinsic instability of LCL. In summary, both the analysis of {gamma}-H2AX and the evaluation of chromosome damage highlighted a larger inter-experimental variability in the results obtained with LCL compared to PBMC. Noteworthy, the two set of results proved to lack any significant correlation at the individual level. These results indicate that LCL may be unsuitable for investigating genotype

  15. Cell-Free DNA in Metastatic Colorectal Cancer

    DEFF Research Database (Denmark)

    Spindler, Karen-Lise G; Boysen, Anders K; Pallisgård, Niels

    2017-01-01

    -analysis of the prognostic value of total cfDNA in patients with metastatic colorectal cancer (mCRC) treated with chemotherapy. In addition, we report on the overall performance of cfDNA as source for KRAS mutation detection. MATERIALS AND METHODS: A systematic literature search of PubMed and Embase was performed by two...... therapy. Small fragments of circulating cell-free DNA (cfDNA) can be measured in a simple blood sample. This report presents the first meta-analysis of the prognostic value of total cfDNA measurement in patients with metastatic colorectal cancer. Data from 1,076 patients confirmed that patients...... with the lowest pre-treatment levels of cfDNA had a significantly higher chance of longer survival than those with higher levels. Cell-free DNA analysis can also be used for detection of tumor-specific mutations, and hold potential as a valuable tool in colorectal cancer treatment....

  16. Physiochemical Characterization of Lignocellulosic Biomass Dissolution by Flowthrough Pretreatment

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Lishi; Pu, Yunqiao; Bowden, Mark; Ragauskas, Arthur J.; Yang, Bin

    2016-01-04

    Comprehensive understanding of biomass solubilization chemistry in aqueous pretreatment such as water-only and dilute acid flowthrough pretreatment is of fundamental importance to achieve the goal of valorizing biomass to fermentable sugars and lignin for biofuels production. In this study, poplar wood was flowthrough pretreated by water-only or 0.05% (w/w) sulfuric acid at different temperatures (220-270 °C), flow rate (25 mL/min), and reaction times (8-90 min), resulting in significant disruption of the lignocellulosic biomass. Ion chromatography (IC), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) analysis, and solid state cross-polarization/magic angle spinning (CP/MAS) 13C nuclear magnetic resonance (NMR) spectroscopy were applied to characterize the pretreated biomass whole slurries in order to reveal depolymerization as well as solubilization mechanism and identify unique dissolution structural features during these pretreatments. Results showed temperature-dependent cellulose decrystallization in flowthrough pretreatment. Crystalline cellulose was completely disrupted, and mostly converted to amorphous cellulose and oligomers by water-only operation at 270 °C for 10 min and by 0.05 wt % H2SO4 flowthrough pretreatment at 220 °C for 12 min. Flowthrough pretreatment with 0.05% (w/w) H2SO4 led to a greater disruption of structures in pretreated poplar at a lower temperature compared to water-only pretreatment.

  17. Pretreatment on corn stover with low concentration of formic acid.

    Science.gov (United States)

    Xu, Jian; Thomsen, Mette Hedegaard; Thomsen, Anne Belinda

    2009-08-01

    Bioethanol derived from lignocellulosic biomass has the potential to replace gasoline. Cellulose is naturally recalcitrant to enzymatic attack, and it also surrounded by the matrix of xylan and lignin, which enhances the recalcitrance. Therefore, lignocellulosic materials must be pretreated to make the cellulose easily degraded into sugars and further fermented to ethanol. In this work, hydrothermal pretreatment on corn stover at 195 degrees for 15 min with and without lower concentration of formic acid was compared in terms of sugar recoveries and ethanol fermentation. For pretreatment with formic acid, the overall glucan recovery was 89% and pretreatment without formic acid yielded the recovery of 94%. Compared with glucan, xylan was more sensitive to the pretreatment condition. The lowest xylan recovery of 55% was obtained after pretreatment with formic acid and the highest of 75% found following pretreatment without formic acid. Toxicity tests of liquor parts showed that there were no inhibitions found for both pretreatment conditions. After simultaneous saccharification and fermentation (SSF) of the pretreated corn stover with Baker's yeast, the highest ethanol yield of 76.5% of the theoretical was observed from corn stover pretreated at 195 degrees for 15 min with formic acid.

  18. DNA probes

    International Nuclear Information System (INIS)

    Castelino, J.

    1992-01-01

    The creation of DNA probes for detection of specific nucleotide segments differs from ligand detection in that it is a chemical rather than an immunological reaction. Complementary DNA or RNA is used in place of the antibody and is labelled with 32 P. So far, DNA probes have been successfully employed in the diagnosis of inherited disorders, infectious diseases, and for identification of human oncogenes. The latest approach to the diagnosis of communicable and parasitic infections is based on the use of deoxyribonucleic acid (DNA) probes. The genetic information of all cells is encoded by DNA and DNA probe approach to identification of pathogens is unique because the focus of the method is the nucleic acid content of the organism rather than the products that the nucleic acid encodes. Since every properly classified species has some unique nucleotide sequences that distinguish it from every other species, each organism's genetic composition is in essence a finger print that can be used for its identification. In addition to this specificity, DNA probes offer other advantages in that pathogens may be identified directly in clinical specimens

  19. Rheological evaluation of pretreated cladding removal waste

    International Nuclear Information System (INIS)

    McCarthy, D.; Chan, M.K.C.; Lokken, R.O.

    1986-01-01

    Cladding removal waste (CRW) contains concentrations of transuranic (TRU) elements in the 80 to 350 nCi/g range. This waste will require pretreatment before it can be disposed of as glass or grout at Hanford. The CRW will be pretreated with a rare earth strike and solids removal by centrifugation to segregate the TRU fraction from the non-TRU fraction of the waste. The centrifuge centrate will be neutralized with sodium hydroxide. This neutralized cladding removal waste (NCRW) is expected to be suitable for grouting. The TRU solids removed by centrifugation will be vitrified. The goal of the Rheological Evaluation of Pretreated Cladding Removal Waste Program was to evaluate those rheological and transport properties critical to assuring successful handling of the NCRW and TRU solids streams and to demonstrate transfers in a semi-prototypic pumping environment. This goal was achieved by a combination of laboratory and pilot-scale evaluations. The results obtained during these evaluations were correlated with classical rheological models and scaled-up to predict the performance that is likely to occur in the full-scale system. The Program used simulated NCRW and TRU solid slurries. Rockwell Hanford Operations (Rockwell) provided 150 gallons of simulated CRW and 5 gallons of simulated TRU solid slurry. The simulated CRW was neutralized by Pacific Northwest Laboratory (PNL). The physical and rheological properties of the NCRW and TRU solid slurries were evaluated in the laboratory. The properties displayed by NCRW allowed it to be classified as a pseudoplastic or yield-pseudoplastic non-Newtonian fluid. The TRU solids slurry contained very few solids. This slurry exhibited the properties associated with a pseudoplastic non-Newtonian fluid

  20. Energy consumption modeling during dairy sewage pretreatment

    Science.gov (United States)

    Dąbrowski, Wojciech; Żyłka, Radosław; Malinowski, Paweł; Boruszko, Dariusz

    2017-11-01

    The research was conducted in a dairy WWTP located in north-eastern Poland with the average flow of 546 m3d-1 and PE 11500 in 2016. Energy consumption was measured with the help of Lumel 3-phase network parameter transducers installed within the plant. The modeling was conducted based on the quantity and quality of raw sewage, after its screening, averaging and dissolved air flotation. The following parameters were determined: BOD5, COD. N-total and P-total. During the research period. 15 measurement series were carried out. Pollution loads removed in primary treatment varied from 167.0 to 803.5 kgO2d-1 and 1205.9 to 10032 kgO2d-1 for BOD5 and COD respectively. The energy consumption share during dairy pretreatment in relation to the total energy consumption was in the range from 13.8 to 28.5% with the mean value of 18.7% during the research period. Energy consumption indicators relating to removed pollution loads for primary treatment were established with the mean values of 0.74 and 0.83 kWhkg-1d-1 for BOD5 and COD respectively. An attempt was made to determine the influence of raw sewage characteristics and pretreatment efficiency on energy consumption of the object. A model of energy consumption during pretreatment was estimated according to the experimental data obtained in the research period. It was modeled using the linear regression model and principal component analysis.

  1. Energy consumption modeling during dairy sewage pretreatment

    Directory of Open Access Journals (Sweden)

    Dąbrowski Wojciech

    2017-01-01

    Full Text Available The research was conducted in a dairy WWTP located in north-eastern Poland with the average flow of 546 m3d-1 and PE 11500 in 2016. Energy consumption was measured with the help of Lumel 3-phase network parameter transducers installed within the plant. The modeling was conducted based on the quantity and quality of raw sewage, after its screening, averaging and dissolved air flotation. The following parameters were determined: BOD5, COD. N-total and P-total. During the research period. 15 measurement series were carried out. Pollution loads removed in primary treatment varied from 167.0 to 803.5 kgO2d-1 and 1205.9 to 10032 kgO2d-1 for BOD5 and COD respectively. The energy consumption share during dairy pretreatment in relation to the total energy consumption was in the range from 13.8 to 28.5% with the mean value of 18.7% during the research period. Energy consumption indicators relating to removed pollution loads for primary treatment were established with the mean values of 0.74 and 0.83 kWhkg-1d-1 for BOD5 and COD respectively. An attempt was made to determine the influence of raw sewage characteristics and pretreatment efficiency on energy consumption of the object. A model of energy consumption during pretreatment was estimated according to the experimental data obtained in the research period. It was modeled using the linear regression model and principal component analysis.

  2. Surface pretreatments for medical application of adhesion

    Directory of Open Access Journals (Sweden)

    Weber Michael

    2003-09-01

    Full Text Available Abstract Medical implants and prostheses (artificial hips, tendono- and ligament plasties usually are multi-component systems that may be machined from one of three material classes: metals, plastics and ceramics. Typically, the body-sided bonding element is bone. The purpose of this contribution is to describe developments carried out to optimize the techniques , connecting prosthesis to bone, to be joined by an adhesive bone cement at their interface. Although bonding of organic polymers to inorganic or organic surfaces and to bone has a long history, there remains a serious obstacle in realizing long-term high-bonding strengths in the in vivo body environment of ever present high humidity. Therefore, different pretreatments, individually adapted to the actual combination of materials, are needed to assure long term adhesive strength and stability against hydrolysis. This pretreatment for metal alloys may be silica layering; for PE-plastics, a specific plasma activation; and for bone, amphiphilic layering systems such that the hydrophilic properties of bone become better adapted to the hydrophobic properties of the bone cement. Amphiphilic layering systems are related to those developed in dentistry for dentine bonding. Specific pretreatment can significantly increase bond strengths, particularly after long term immersion in water under conditions similar to those in the human body. The bond strength between bone and plastic for example can be increased by a factor approaching 50 (pealing work increasing from 30 N/m to 1500 N/m. This review article summarizes the multi-disciplined subject of adhesion and adhesives, considering the technology involved in the formation and mechanical performance of adhesives joints inside the human body.

  3. Enzymatic Hydrolysis of Alkaline Pretreated Coconut Coir

    Directory of Open Access Journals (Sweden)

    Akbarningrum Fatmawati

    2013-06-01

    Full Text Available The purpose of this research is to study the effect of concentration and temperature on the cellulose and lignin content, and the reducing sugars produced in the enzymatic hydrolysis of coconut coir. In this research, the coconut coir is pretreated using 3%, 7%, and 11% NaOH solution at 60oC, 80oC, and 100oC. The pretreated coir were assayed by measuring the amount of cellulose and lignin and then hydrolysed using Celluclast and Novozyme 188 under various temperature (30oC, 40oC, 50oC and pH (3, 4, 5. The hydrolysis results were assayed for the reducing sugar content. The results showed that the alkaline delignification was effective to reduce lignin and to increase the cellulose content of the coir. The best delignification condition was observed at 11% NaOH solution and 100oC which removed 14,53% of lignin and increased the cellulose content up to 50,23%. The best condition of the enzymatic hydrolysis was obtained at 50oC and pH 4 which produced 7,57 gr/L reducing sugar. © 2013 BCREC UNDIP. All rights reservedReceived: 2nd October 2012; Revised: 31st January 2013; Accepted: 6th February 2013[How to Cite: Fatmawati, A., Agustriyanto, R., Liasari, Y. (2013. Enzymatic Hydrolysis of Alkaline Pre-treated Coconut Coir. Bulletin of Chemical Reaction Engineering & Catalysis, 8 (1: 34-39 (doi:10.9767/bcrec.8.1.4048.34-39[Permalink/DOI: http://dx.doi.org/10.9767/bcrec.8.1.4048.34-39] | View in  |

  4. Acetic acid pretreatment improves the hardness of cooked potato slices.

    Science.gov (United States)

    Zhao, Wenlin; Shehzad, Hussain; Yan, Shoulei; Li, Jie; Wang, Qingzhang

    2017-08-01

    The effects of acetic acid pretreatment on the texture of cooked potato slices were investigated in this work. Potato slices were pretreated with acetic acid immersion (AAI), distilled water immersion (DWI), or no immersion (NI). Subsequently, the cell wall material of the pretreated samples was isolated and fractioned to evaluate changes in the monosaccharide content and molar mass (MM), and the hardness and microscopic structure of the potato slices in different pretreatments before and after cooking were determined. The results showed that the highest firmness was obtained with more intact structure of the cell wall for cooked potato slices with AAI pretreatment. Furthermore, the MM and sugar ratio demonstrated that the AAI pretreated potato slices contained a higher content of the small molecular polysaccharides of cell walls, especially in the hemicellulose fraction. This work may provide a reference for potato processing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Impact of pretreatment on colour and texture of watermelon rind

    Science.gov (United States)

    Athmaselvi, K.; Alagusundaram, K.; Kavitha, C.; Arumuganathan, T.

    2012-07-01

    The effect of osmotic dehydration pretreatment on water loss, solid gain, colour and textural change was investigated. Watermelon rind 1 x 1 cm size was immersed in sucrose solution of 40, 50 and 60° Brix after pretreatment with microwave and conventional boiling in water for 1, 3, and 5 min, respectively. Water loss and solid gain increased with the time of cooking and sugar concentration. Microwave pretreated samples showed higher water loss and solid gain. Increase in the time of cooking decreased the brightness of all the samples. Microwave pretreated samples showed higher `b' values than conventionally pretreated ones. There was no significant difference (P≤0.05) in texture profile analysis parameters except for hardness. Hardness decreased with increase in time of cooking and sugar concentration. Second order regression model was developed for water loss and solid gain of microwave and conventional pretreated watermelon rind.

  6. DNA data

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Raw DNA chromatogram data produced by the ABI 373, 377, 3130 and 3730 automated sequencing machines in ABI format. These are from fish (primarily Sebastes spp.,...

  7. DNA methylation

    DEFF Research Database (Denmark)

    Williams, Kristine; Christensen, Jesper; Helin, Kristian

    2012-01-01

    DNA methylation is involved in key cellular processes, including X-chromosome inactivation, imprinting and transcriptional silencing of specific genes and repetitive elements. DNA methylation patterns are frequently perturbed in human diseases such as imprinting disorders and cancer. The recent...... discovery that the three members of the TET protein family can convert 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) has provided a potential mechanism leading to DNA demethylation. Moreover, the demonstration that TET2 is frequently mutated in haematopoietic tumours suggests that the TET...... proteins are important regulators of cellular identity. Here, we review the current knowledge regarding the function of the TET proteins, and discuss various mechanisms by which they contribute to transcriptional control. We propose that the TET proteins have an important role in regulating DNA methylation...

  8. DNA nanotechnology

    Science.gov (United States)

    Seeman, Nadrian C.; Sleiman, Hanadi F.

    2018-01-01

    DNA is the molecule that stores and transmits genetic information in biological systems. The field of DNA nanotechnology takes this molecule out of its biological context and uses its information to assemble structural motifs and then to connect them together. This field has had a remarkable impact on nanoscience and nanotechnology, and has been revolutionary in our ability to control molecular self-assembly. In this Review, we summarize the approaches used to assemble DNA nanostructures and examine their emerging applications in areas such as biophysics, diagnostics, nanoparticle and protein assembly, biomolecule structure determination, drug delivery and synthetic biology. The introduction of orthogonal interactions into DNA nanostructures is discussed, and finally, a perspective on the future directions of this field is presented.

  9. Lignocellulosic biomass-Thermal pretreatment with steam: Pretreatment techniques for biofuels and biorefineries

    DEFF Research Database (Denmark)

    Toor, Saqib; Rosendahl, Lasse; Hoffmann, Jessica

    2013-01-01

    technologies; thermal pre-treatment with steam. The Norwegian company CAMBI developed a process for treatment of sludge from waste water treatment plants, and the idea was based on the experience that cooking sludge under pressure at temperature from 150oC to 180oC improved the digestibility and at the same...

  10. The characteristic changes of betung bamboo (Dendrocalamus asper pretreated by fungal pretreatment

    Directory of Open Access Journals (Sweden)

    Widya Fatriasari

    2014-05-01

    Full Text Available Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 The fungal pretreatment effect on chemical structural and morphological changes of Betung Bamboo was evaluated based on its biomass components after being cultivated by white rot fungi, Trametes versicolor. Betung bamboo powder (15 g was exposed to liquid inoculum of white rot fungi and incubated at 270C for 15, 30 and 45 days. The treated samples were then characterized by FT-IR spectroscopy, X-Ray diffraction and SEM-EDS analyses. Cultivation for 30 days with 5 and 10% loadings retained greater selectivity compared to that of the other treatments. FTIR spectra demonstrated that the fungus affected the decreasing of functional group quantities without changing the functional groups. The decrease in intensity at wave number of 1246 cm-1 (guaiacyl of lignin was greater than that at wave number of 1328 cm-1 (deformation combination of syringyl and xylan after fungal treatment. X-ray analysis showed the pretreated samples had a higher crystallinity than the untreated ones which might be due to the cleavage of amorphous fractions of cellulose. The pretreated samples have more fragile than the untreated ones confirmed by SEM. Crystalline allomorph calculated by XRD analysis showed that fungus pretreatment for 30 days has transformed triclinic structure of cellulose to monoclinic structure.

  11. Influence of caffeine pretreatment on biphasic in vitro maturation of dog oocytes.

    Science.gov (United States)

    Salavati, Mazdak; Ghafari, Fataneh; Zhang, Tiantian; Fouladi-Nashta, Ali A

    2013-10-15

    Phosphodiesterase (PDE) inhibitors have been utilized for in vitro maturation (IVM) of oocytes to manipulate the meiotic resumption and progression. Premature chromatin condensation and DNA replication of the oocytes, immediately after the decrease in the cAMP level, are the difficulties in canine IVM. Caffeine, a nonselective competitive PDE inhibitor, due to its structural similarity to adenosine molecule maintains the cAMP level by occupying PDE enzymes such as PDE-3A inside the oocyte and PDE-4 and PDE-5 in the cumulus cells. In this study, the effects of 12-hour caffeine pretreatment in a biphasic IVM protocol were assessed on maturation rates of canine oocytes. Sixty hours of culture after a 12-hour of 10 mM caffeine pretreatment resulted in 16.9% ± 2.4 of the oocytes reaching metaphase II stage (MII) and 25.9% ± 5.2 degeneration rate compared with the control group with 2.2% ± 2.2 MII and 37.6% ± 4.3 degeneration rates (P BCB) staining. Oocytes showed difference in meiotic resumption (MI-MII) (BCB+ = 16.11% ± 5.5, BCB- = 9.86% ± 5.0; P BCB+ canine oocytes had higher MII rate than the BCB- group under caffeine pretreatment (10.2% ± 2.9 vs. 1.1% ± 1.1, respectively; P BCB+ oocytes have higher competency in vitro for nuclear maturation. Copyright © 2013 Elsevier Inc. All rights reserved.

  12. FRACTIONAL CRYSTALLIZATION TESTING WITH INTERIM PRETREATMENT SYSTEM FEEDS

    International Nuclear Information System (INIS)

    HERTING DL

    2008-01-01

    The fractional crystallization process was developed as a pretreatment method for saltcake waste retrieved from Hanford single-shell tanks (SST). The process separates the retrieved SST waste into a high-level waste stream containing the bulk of the radionuclides and a low-activity waste stream containing the bulk of the nonradioactive sodium salts. The Interim Pretreatment System project shifted the focus on pretreatment planning from SST waste to double-shell tank waste

  13. Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities

    Directory of Open Access Journals (Sweden)

    Fei Cheng

    Full Text Available Abstract Humic substances in soil DNA samples can influence the assessment of microbial diversity and community composition. Using multiple steps during or after cell lysis adds expenses, is time-consuming, and causes DNA loss. A pretreatment of soil samples and a single step DNA extraction may improve experimental results. In order to optimize a protocol for obtaining high purity DNA from soil microbiota, five prewashing agents were compared in terms of their efficiency and effectiveness in removing soil contaminants. Residual contaminants were precipitated by adding 0.6 mL of 0.5 M CaCl2. Four cell lysis methods were applied to test their compatibility with the pretreatment (prewashing + Ca2+ flocculation and to ultimately identify the optimal cell lysis method for analyzing fungal communities in forest soils. The results showed that pretreatment with TNP + Triton X-100 + skim milk (100 mM Tris, 100 mM Na4P2O7, 1% polyvinylpyrrolidone, 100 mM NaCl, 0.05% Triton X-100, 4% skim milk, pH 10.0 removed most soil humic contaminants. When the pretreatment was combined with Ca2+ flocculation, the purity of all soil DNA samples was further improved. DNA samples obtained by the fast glass bead-beating method (MethodFGB had the highest purity. The resulting DNA was successfully used, without further purification steps, as a template for polymerase chain reaction targeting fungal internal transcribed spacer regions. The results obtained by terminal restriction fragment length polymorphism analysis indicated that the MethodFGB revealed greater fungal diversity and more distinctive community structure compared with the other methods tested. Our study provides a protocol for fungal cell lysis in soil, which is fast, convenient, and effective for analyzing fungal communities in forest soils.

  14. Cellulose conversion of corn pericarp without pretreatment.

    Science.gov (United States)

    Kim, Daehwan; Orrego, David; Ximenes, Eduardo A; Ladisch, Michael R

    2017-12-01

    We report enzyme hydrolysis of cellulose in unpretreated pericarp at a cellulase loading of 0.25FPU/g pericarp solids using a phenol tolerant Aspergillus niger pectinase preparation. The overall protein added was 5mg/g and gave 98% cellulose conversion in 72h. However, for double the amount of enzyme from Trichoderma reesei, which is significantly less tolerant to phenols, conversion was only 16%. The key to achieving high conversion without pretreatment is combining phenol inhibition-resistant enzymes (such as from A. niger) with unground pericarp from which release of phenols is minimal. Size reduction of the pericarp, which is typically carried out in a corn-to-ethanol process, where corn is first ground to a fine powder, causes release of highly inhibitory phenols that interfere with cellulase enzyme activity. This work demonstrates hydrolysis without pretreatment of large particulate pericarp is a viable pathway for directly producing cellulose ethanol in corn ethanol plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. DNA expressions - A formal notation for DNA

    NARCIS (Netherlands)

    Vliet, Rudy van

    2015-01-01

    We describe a formal notation for DNA molecules that may contain nicks and gaps. The resulting DNA expressions denote formal DNA molecules. Different DNA expressions may denote the same molecule. Such DNA expressions are called equivalent. We examine which DNA expressions are minimal, which

  16. Memory CD8(+) T cells elicited by HIV-1 lipopeptide vaccines display similar phenotypic profiles but differences in term of magnitude and multifunctionality compared with FLU- or EBV-specific memory T cells in humans.

    Science.gov (United States)

    Figueiredo, Suzanne; Charmeteau, Benedicte; Surenaud, Mathieu; Salmon, Dominique; Launay, Odile; Guillet, Jean-Gérard; Hosmalin, Anne; Gahery, Hanne

    2014-01-16

    Differentiation marker, multifunctionality and magnitude analyses of specific-CD8(+) memory T cells are crucial to improve development of HIV vaccines designed to generate cell-mediated immunity. Therefore, we fully characterized the HIV-specific CD8(+) T cell responses induced in volunteers vaccinated with HIV lipopeptide vaccines for phenotypic markers, tetramer staining, cytokine secretion, and cytotoxic activities. The frequency of ex vivo CD8(+) T cells elicited by lipopeptide vaccines is very rare and central-memory phenotype and functions of these cells were been shown to be important in AIDS immunity. So, we expanded them using specific peptides to compare the memory T cell responses induced in volunteers by HIV vaccines with responses to influenza (FLU) or Epstein Barr virus (EBV). By analyzing the differentiation state of IFN-γ-secreting CD8(+) T cells, we found a CCR7(-)CD45RA(-)CD28(+int)/CD28(-) profile (>85%) belonging to a subset of intermediate-differentiated effector T cells for HIV, FLU, and EBV. We then assessed the quality of the response by measuring various T cell functions. The percentage of single IFN-γ T cell producers in response to HIV was 62% of the total of secreting T cells compared with 35% for FLU and EBV, dual and triple (IFN-γ/IL-2/CD107a) T cell producers could also be detected but at lower levels (8% compared with 37%). Finally, HIV-specific T cells secreted IFN-γ and TNF-α, but not the dual combination like FLU- and EBV-specific T cells. Thus, we found that the functional profile and magnitude of expanded HIV-specific CD8(+) T precursors were more limited than those of to FLU- and EBV-specific CD8(+) T cells. These data show that CD8(+) T cells induced by these HIV vaccines have a similar differentiation profile to FLU and EBV CD8(+) T cells, but that the vaccine potency to induce multifunctional T cells needs to be increased in order to improve vaccination strategies. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Biologically Pre-Treated Habitation Waste Water as a Sustainable Green Urine Pre-Treat Solution

    Science.gov (United States)

    Jackson, W. Andrew; Thompson, Bret; Sevanthi, Ritesh; Morse, Audra; Meyer, Caitlin; Callahan, Michael

    2017-01-01

    The ability to recover water from urine and flush water is a critical process to allow long term sustainable human habitation in space or bases on the moon or mars. Organic N present as urea or similar compounds can hydrolyze producing free ammonia. This reaction results in an increase in the pH converting ammonium to ammonia which is volatile and not removed by distillation. The increase in pH will also cause precipitation reactions to occur. In order to prevent this, urine on ISS is combined with a pretreat solution. While use of a pretreatment solution has been successful, there are numerous draw backs including: storage and use of highly hazardous solutions, limitations on water recovery (less than 85%), and production of brine with pore dewatering characteristics. We evaluated the use of biologically treated habitation wastewaters (ISS and early planetary base) to replace the current pretreat solution. We evaluated both amended and un-amended bioreactor effluent. For the amended effluent, we evaluated "green" pretreat chemicals including citric acid and citric acid amended with benzoic acid. We used a mock urine/air separator modeled after the urine collection assembly on ISS. The urine/air separator was challenged continually for >6 months. Depending on the test point, the separator was challenged daily with donated urine and flushed with amended or un-amended reactor effluent. We monitored the pH of the urine, flush solution and residual pH in the urine/air separator after each urine event. We also evaluated solids production and biological growth. Our results support the use of both un-amended and amended bioreactor effluent to maintain the operability of the urine /air separator. The ability to use bioreactor effluent could decrease consumable cost, reduce hazards associated with current pre-treat chemicals, allow other membrane based desalination processes to be utilized, and improve brine characteristics.

  18. Feasibility of Hydrothermal Pretreatment on Maize Silage for Bioethanol Production

    DEFF Research Database (Denmark)

    Xu, Jian; Thomsen, Mette Hedegaard; Thomsen, Anne Belinda

    2010-01-01

    The potential of maize silage as a feedstock to produce bioethanol was evaluated in the present study. The hydrothermal pretreatment with five different pretreatment severity factors (PSF) was employed to pretreat the maize silage and compared in terms of sugar recovery, toxic test, and ethanol...... the liquors from the five conditions were not toxic to the Baker’s yeast. Pretreatment under 195°C for 7 min had the similar PSF with that of 185°C for 15 min, and both gave the higher ethanol concentration of 19.92 and 19.98 g/L, respectively. The ethanol concentration from untreated maize silage was only 7...

  19. Nonhazardous Urine Pretreatment Method for Future Exploration Systems, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — A nonhazardous urine pretreatment system prototype is proposed that will stabilize urine against biological growth or chemical instabilities without using hazardous...

  20. Nonhazardous Urine Pretreatment Method for Future Exploration Systems, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — A novel urine pretreatment that will prevent biological growth or chemical instabilities in urine without using hazardous chemicals is proposed. Untreated urine...

  1. What Is Mitochondrial DNA?

    Science.gov (United States)

    ... DNA What is mitochondrial DNA? What is mitochondrial DNA? Although most DNA is packaged in chromosomes within ... proteins. For more information about mitochondria and mitochondrial DNA: Molecular Expressions, a web site from the Florida ...

  2. Saccharification and ethanol fermentation from cholinium ionic liquid-pretreated bagasse with a different number of post-pretreatment washings.

    Science.gov (United States)

    Ninomiya, Kazuaki; Omote, Sayuri; Ogino, Chiaki; Kuroda, Kosuke; Noguchi, Mana; Endo, Takatsugu; Kakuchi, Ryohei; Shimizu, Nobuaki; Takahashi, Kenji

    2015-01-01

    Choline acetate (ChOAc), a cholinium ionic liquid (IL), was compared with 1-ethyl-3-methylimidazolium acetate (EmimOAc) with regard to biomass pretreatment, inhibition on cellulase and yeast, residuals in pretreated biomass, and saccharification and fermentation of pretreated biomass. Irrespective of ChOAc and EmimOAc, cellulose and hemicellulose saccharification of the IL-pretreated bagasse were over 90% and 60%, respectively. Median effective concentrations (EC50) based on cellulase activity were 32 wt% and 16 wt% for ChOAc and EmimOAc, respectively. The EC50 based on yeast growth were 3.1 wt% and 0.3 wt% for ChOAc and EmimOAc respectively. The residuals in IL-pretreated bagasse were 10% and 23% for ChOAc and EmimOAc, respectively, when washed 2 times after pretreatment. Ethanol yield on a bagasse basis were 60% and 24% for ChOAc and EmimOAc, respectively, in the saccharification and fermentation of IL-pretreated bagasse when washed 2 times. ChOAc-pretreated bagasse could be saccharified and fermented with fewer wash times than EmimOAc-pretreated bagasse. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Glycerol carbonate as green solvent for pretreatment of sugarcane bagasse

    Science.gov (United States)

    2013-01-01

    Background Pretreatment of lignocellulosic biomass is a prerequisite for effective saccharification to produce fermentable sugars. In this study, “green” solvent systems based on acidified mixtures of glycerol carbonate (GC) and glycerol were used to treat sugarcane bagasse and the roles of each solvent in deconstructing biomass were determined. Results Pretreatment of sugarcane bagasse at 90°C for only 30 min with acidified GC produced a solid residue having a glucan digestibility of 90% and a glucose yield of 80%, which were significantly higher than a glucan digestibility of 16% and a glucose yield of 15% obtained for bagasse pretreated with acidified ethylene carbonate (EC). Biomass compositional analyses showed that GC pretreatment removed more lignin than EC pretreatment (84% vs 54%). Scanning electron microscopy (SEM) showed that fluffy and size-reduced fibres were produced from GC pretreatment whereas EC pretreatment produced compact particles of reduced size. The maximal glucan digestibility and glucose yield of GC/glycerol systems were about 7% lower than those of EC/ethylene glycol (EG) systems. Replacing up to 50 wt% of GC with glycerol did not negatively affect glucan digestibility and glucose yield. The results from pretreatment of microcrystalline cellulose (MCC) showed that (1) pretreatment with acidified alkylene glycol (AG) alone increased enzymatic digestibility compared to pretreatments with acidified alkylene carbonate (AC) alone and acidified mixtures of AC and AG, (2) pretreatment with acidified GC alone slightly increased, but with acidified EC alone significantly decreased, enzymatic digestibility compared to untreated MCC, and (3) there was a good positive linear correlation of enzymatic digestibility of treated and untreated MCC samples with congo red (CR) adsorption capacity. Conclusions Acidified GC alone was a more effective solvent for pretreatment of sugarcane bagasse than acidified EC alone. The higher glucose yield obtained

  4. Understanding of alkaline pretreatment parameters for corn stover enzymatic saccharification

    Directory of Open Access Journals (Sweden)

    Chen Ye

    2013-01-01

    Full Text Available Abstract Background Previous research on alkaline pretreatment has mainly focused on optimization of the process parameters to improve substrate digestibility. To achieve satisfactory sugar yield, extremely high chemical loading and enzyme dosages were typically used. Relatively little attention has been paid to reduction of chemical consumption and process waste management, which has proven to be an indispensable component of the bio-refineries. To indicate alkali strength, both alkali concentration in pretreatment solution (g alkali/g pretreatment liquor or g alkali/L pretreatment liquor and alkali loading based on biomass solids (g alkali/g dry biomass have been widely used. The dual approaches make it difficult to compare the chemical consumption in different process scenarios while evaluating the cost effectiveness of this pretreatment technology. The current work addresses these issues through pretreatment of corn stover at various combinations of pretreatment conditions. Enzymatic hydrolysis with different enzyme blends was subsequently performed to identify the effects of pretreatment parameters on substrate digestibility as well as process operational and capital costs. Results The results showed that sodium hydroxide loading is the most dominant variable for enzymatic digestibility. To reach 70% glucan conversion while avoiding extensive degradation of hemicellulose, approximately 0.08 g NaOH/g corn stover was required. It was also concluded that alkali loading based on total solids (g NaOH/g dry biomass governs the pretreatment efficiency. Supplementing cellulase with accessory enzymes such as α-arabinofuranosidase and β-xylosidase significantly improved the conversion of the hemicellulose by 6–17%. Conclusions The current work presents the impact of alkaline pretreatment parameters on the enzymatic hydrolysis of corn stover as well as the process operational and capital investment costs. The high chemical consumption for alkaline

  5. Evaluation of High Solids Alkaline Pretreatment of Rice Straw

    Science.gov (United States)

    Cheng, Yu-Shen; Zheng, Yi; Yu, Chao Wei; Dooley, Todd M.; Jenkins, Bryan M.

    2010-01-01

    Fresh-harvested, air-dried rice straw was pretreated at a water content of 5 g H2O/g straw using sodium hydroxide (NaOH) and compared to pretreatment at 10 g H2O/g straw by hydrated lime (Ca(OH)2). Full factorial experiments including parallel wash-only treatments were completed with both sources of alkali. The experiments were designed to measure the effects of alkaline loading and pretreatment time on delignification and sugar yield upon enzymatic hydrolysis. Reaction temperature was held constant at 95°C for lime pretreatment and 55°C for NaOH pretreatment. The range of delignification was 13.1% to 27.0% for lime pretreatments and was 8.6% to 23.1% for NaOH pretreatments. Both alkaline loading and reaction time had significant positive effects (p delignification under the design conditions, but only alkaline loading had a significant positive effect on enzymatic hydrolysis. Treatment at higher temperature also improved delignification; delignification with water alone ranged from 9.9% to 14.5% for pretreatment at 95°C, but there was little effect observed at 55°C. Post-pretreatment washing of biomass was not necessary for subsequent enzymatic hydrolysis. Maximum glucose yields were 176.3 mg/g dried biomass (48.5% conversion efficiency of total glucose) in lime-pretreated and unwashed biomass and were 142.3 mg/g dried biomass (39.2% conversion efficiency of total glucose) in NaOH-pretreated and unwashed biomass. PMID:20440580

  6. Quantitation by flow microfluorometry of total cellular DNA in Acanthamoeba

    Energy Technology Data Exchange (ETDEWEB)

    Coulson, P.B.; Tyndall, R.

    1978-01-01

    The DNA content of five speciea of Acanthamoeba was determined by flow microfluorometry. Acanthamoeba castellanii (AC-30), acanthamoeba polyphaga (APG and P-23), acanthamoeba rhysodes, acanthamoeba culbertsoni (A-1), and acanthamoeba royreba were grown in a casitone based medium 24 to 48 hr. The trophozoites were harvested, fixed in 70% ethanol (acidified), pretreated with RNase, stained with propidium diiodide, and evaluated for DNA-bound fluorescence. All species tested had DNA values between 2.0 to 5.0 pg/cell. These results placed DNA/cell values of Acanthamoeba slightly lower than DNA/cell values of other eucaryotic cells and much lower than Amoeba proteus values. These results indicate that FMF may be a useful adjunct in distinguishing Acanthamoeba cells from either eucaryotic cells or some other amoeba. However, differences in DNA/cell between species of Acanthamoeba are small and would not be useful in identification of species.

  7. Plasma-Assisted Pretreatment of Wheat Straw

    DEFF Research Database (Denmark)

    Schultz-Jensen, Nadja; Leipold, Frank; Bindslev, Henrik

    2011-01-01

    O3 generated in a plasma at atmospheric pressure and room temperature, fed with dried air (or oxygen-enriched dried air), has been used for the degradation of lignin in wheat straw to optimize the enzymatic hydrolysis and to get more fermentable sugars. A fixed bed reactor was used combined...... with a CO2 detector and an online technique for O3 measurement in the fed and exhaust gas allowing continuous measurement of the consumption of O3. This rendered it possible for us to determine the progress of the pretreatment in real time (online analysis). The process time can be adjusted to produce wheat...... straw with desired lignin content because of the online analysis. The O3 consumption of wheat straw and its polymeric components, i.e., cellulose, hemicellulose, and lignin, as well as a mixture of these, dry as well as with 50% water, were studied. Furthermore, the process parameters dry matter content...

  8. Analysis of ancient DNA from coprolites: a perspective with random amplified polymorphic DNA-polymerase chain reaction approach

    Directory of Open Access Journals (Sweden)

    Iñiguez Alena M

    2003-01-01

    Full Text Available The aim of this work was to determine approaches that would improve the quality of ancient DNA (aDNA present in coprolites to enhance the possibility of success in retrieving specific sequence targets. We worked with coprolites from South American archaeological sites in Brazil and Chile dating up to 7,000 years ago. Using established protocols for aDNA extraction we obtained samples showing high degradation as usually happens with this kind of material. The reconstructive polymerization pretreatment was essential to overcome the DNA degradation and the serial dilutions helped with to prevent polymerase chain reaction (PCR inhibitors. Moreover, the random amplified polymorphic DNA-PCR has been shown to be a reliable technique for further experiments to recover specific aDNA sequences.

  9. DNA nanotechnology

    OpenAIRE

    Nadrian C Seeman

    2003-01-01

    Since Watson and Crick’s determination of its structure nearly 50 years ago, DNA has come to fill our lives in many areas, from genetic counseling to forensics, from genomics to gene therapy. These, and other ways in which DNA affects human activities, are related to its function as genetic material, not just our genetic material, but the genetic material of all living organisms. Here, we will ignore DNA’s biological role; rather, we will discuss how the properties that make it so successful ...

  10. Quality evaluation of beverage produced from pre-treated tigernut ...

    African Journals Online (AJOL)

    Beverage samples from pretreated and untreated tigernut were evaluated for physical and chemical properties. Sensory evaluation of the beverage samples were carried out and the best preferred tigernutbeverage was compared with a popularly consumed beverage, soymilk. All the pre-treatments, except germination, ...

  11. Effects of magnetic fields pretreatment of mungbean seeds on sprout ...

    African Journals Online (AJOL)

    Effects of magnetic fields pretreatment of mungbean seeds on sprout yield and quality. Yi-Ping Chen, Jun-Min He, Ran Li. Abstract. The aim of this investigation was to determine the effect of magnetic field pretreatment of mungbean seeds on the yield and quality of sprout. The sprout elongation, biomass and nutrition ...

  12. Wet explosion pretreatment of sugarcane bagasse for enhanced enzymatic hydrolysis

    DEFF Research Database (Denmark)

    Biswas, Rajib; Uellendahl, Hinrich; Ahring, Birgitte Kiær

    2014-01-01

    Wet explosion pretreatment of sugarcane bagasse was investigated in pilot-scale with the aim of obtaining the highest possible sugar yield after pretreatment. The temperatures used were 155, 170, 185 and 200 C with or without addition of oxygen (0.6 MPa pressure). Enzymatic hydrolysis of washed...

  13. Synergistic Effect of Trehalose and Saccharose Pretreatment on ...

    African Journals Online (AJOL)

    Purpose: To investigate the synergistic effect of trehalose and saccharose pretreatment on maintenance of lyophilized human red blood cell (RBC) quality. Methods: RBCs were pre-treated with trehalose and saccharose, and then lyophilized and re-hydrated. Prior to lyophilization and after re hydration, RBC parameters, ...

  14. 40 CFR 406.26 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 406.26 Section 406.26 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Corn Dry Milling Subcategory § 406.26 Pretreatment...

  15. 40 CFR 406.34 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for existing sources. 406.34 Section 406.34 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... § 406.34 Pretreatment standards for existing sources. Any existing source subject to this subpart that...

  16. 40 CFR 406.44 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for existing sources. 406.44 Section 406.44 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED... § 406.44 Pretreatment standards for existing sources. Any existing source subject to this subpart that...

  17. 40 CFR 406.16 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 406.16 Section 406.16 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Corn Wet Milling Subcategory § 406.16 Pretreatment...

  18. Effect of chemical pretreatment of some lignocellulosic wastes on ...

    African Journals Online (AJOL)

    Highest cellulase activity (0.06777 IU/ml/min) was display d by the organism grown on bagasse substrate pretreated with 2M NaOH for one hour. The proximate analysis of the cellulosic residues differed from one substrate to another, with the bagasse being the best. Pulverized substrates syndicated with alkali pretreatment ...

  19. Influence of Chemical Pretreatment on the Quality of Dried Whole ...

    African Journals Online (AJOL)

    Chemical pretreatment of vegetables prior to drying is widely used to improve colour, texture and overall acceptability. Thus, this study investigated the effect of chemical pretreatment on some physico-chemical properties, colour and rehydration properties of dried red bell pepper. Fresh samples of whole red bell peppers ...

  20. Organosolv pretreatment of olive tree biomass for fermentable sugars

    Energy Technology Data Exchange (ETDEWEB)

    Diaz, M.J.; Cara, C.; Castro, E. [Department of Chemical, Environmental and Materials Engineering, Campus Las Lagunillas, University of Jaen, Jaen (Spain); Huijgen, W.J.J.; Van der Laan, R.R.; Reith, J.H. [ECN Biomass, Coal and Environmental Research, Petten (Netherlands)

    2011-03-15

    Olive tree pruning biomass is one of the main agricultural residues available in Mediterranean countries and is currently lacking commercial applications. To take advantage of its sugar content, a pretreatment is necessary to enhance enzyme accessibility of the cellulose fraction of the residue. This paper describes for the first time the use of organosolv pretreatment in this regard. The main process variables such as pretreatment temperature, residence time, and solvent composition (aqueous ethanol) are studied. Results show that organosolv pretreatment causes delignification and hydrolysis of hemicelluloses and improves the enzymatic digestibility of olive tree pruning biomass. A higher pretreatment severity and ethanol content of the solvent were found to increase delignification (up to 64% at 66% w/w aqueous ethanol, 210C, 60 min). By contrast, xylan hydrolysis was promoted by a lower ethanol content (maximum 92%). The highest enzymatic hydrolysis yield (90% of the structural glucan present in the raw material) has been obtained after pretreatment with 43% w/w aqueous ethanol at 210C for 15 min. Organosolv pretreatment was found to be the most effective pretreatment for enzymatic hydrolysis of olive tree pruning biomass.

  1. Effect of combination pre-treatment on physicochemical, sensory ...

    African Journals Online (AJOL)

    Effect of combination pre-treatment on physicochemical, sensory and microbial characteristics of fresh aerobically stored minced goat (Black Bengal) meat organs. ... African Journal of Biotechnology ... However, acetic acid and glucose pretreatment controlled the fungal growth in meat samples most effectively. The

  2. 40 CFR 427.86 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Pretreatment standards for new sources. 427.86 Section 427.86 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT... Textiles Subcategory § 427.86 Pretreatment standards for new sources. Any new source subject to this...

  3. Alkaline pretreatment of Mexican pine residues for bioethanol ...

    African Journals Online (AJOL)

    CLAUDIA

    2013-07-31

    Jul 31, 2013 ... form of branches and tree tops are left on the field. Part of this residue is cut into ..... Table 3. Chemical composition of the solids from pretreated and hydrolyzed biomass and black and white liquors obtained in the pretreatment during the enzymatic .... (2000) employed Oak wood lignocellulosic biomass ...

  4. Development of pretreatment of empty fruit bunches for enhanced ...

    African Journals Online (AJOL)

    Heating, boiling and steaming are among the physical agents and different concentrations of nitric acid, sulfuric acid and sodium hydroxide (NaOH) were the chemical agents used for the pretreatment of EFB to enhance the enzymatic saccharification of EFB. NaOH was proved to be the best among all the pretreatment ...

  5. Structural Analysis of Alkaline Pretreated Rice Straw for Ethanol Production

    Directory of Open Access Journals (Sweden)

    Paripok Phitsuwan

    2017-01-01

    Full Text Available Rice straw (RS is an abundant, readily available agricultural waste, which shows promise as a potential feedstock for Asian ethanol production. To enhance release of glucose by enzymatic hydrolysis, RS was pretreated with aqueous ammonia (27% w/w at two pretreatment temperatures: room temperature and 60°C. Statistical analysis indicated similarity of enzymatic glucose production at both pretreatment temperatures after 3-day incubation. Chemical composition, FTIR, and EDX analyses confirmed the retention of glucan and xylan in the pretreated solid, but significant reduction of lignin (60.7% removal and silica. SEM analysis showed the disorganized surfaces and porosity of the pretreated RS fibers, thus improving cellulose accessibility for cellulase. The crystallinity index increased from 40.5 to 52.3%, indicating the higher exposure of cellulose. With 10% (w/v solid loadings of pretreated RS, simultaneous saccharification and fermentation yielded a final ethanol concentration of 24.6 g/L, corresponding to 98% of maximum theoretical yield. Taken together, aqueous ammonia pretreatment is an effective method to generate highly digestible pretreated RS for bioethanol production and demonstrates potential application in biorefinery industry.

  6. Essential Role of Rta in Lytic DNA Replication of Epstein-Barr Virus

    Science.gov (United States)

    Ghiassi-Nejad, Maryam; Golden, Sean; Delecluse, Henri-Jacques; Miller, George

    2013-01-01

    Two transcription factors, ZEBRA and Rta, switch Epstein-Barr virus (EBV) from the latent to the lytic state. While ZEBRA also plays an obligatory role as an activator of replication, it is not known whether Rta is directly required for replication. Rta is dispensable for amplification of an oriLyt-containing plasmid in a transient-replication assay. Here, we assessed the requirement for Rta in activation of viral DNA synthesis from the endogenous viral genome, a function that has not been established. Initially, we searched for a ZEBRA mutant that supports viral replication but not transcription. We found that Z(S186A), a mutant of ZEBRA unable to activate transcription of Rta or viral genes encoding replication proteins, is competent to bind to oriLyt and to function as an origin recognition protein. Ectopic expression of the six components of the EBV lytic replication machinery failed to rescue replication by Z(S186A). However, addition of Rta to Z(S186A) and the mixture of replication factors activated viral replication and late gene expression. Deletion mutagenesis of Rta indicated that the C-terminal 10 amino acids (aa) were essential for the function of Rta in replication. In vivo DNA binding studies revealed that Rta interacted with the enhancer region of oriLyt. In addition, expression of Rta and Z(S186A) together, but not individually, activated synthesis of the BHLF1 transcript, a lytic transcript required for the process of viral DNA replication. Our findings demonstrate that Rta plays an indispensable role in the process of lytic DNA replication. PMID:23077295

  7. Hypomethylation and Over-Expression of the Beta Isoform of BLIMP1 is Induced by Epstein-Barr Virus Infection of B Cells; Potential Implications for the Pathogenesis of EBV-Associated Lymphomas

    Directory of Open Access Journals (Sweden)

    Katerina Vrzalikova

    2012-10-01

    Full Text Available B-lymphocyte-induced maturation protein 1 (BLIMP1 exists as two major isoforms, α and β, which arise from alternate promoters. Inactivation of the full length BLIMP1α isoform is thought to contribute to B cell lymphomagenesis by blocking post-germinal centre (GC B cell differentiation. In contrast, the shorter β isoform is functionally impaired and over-expressed in several haematological malignancies, including diffuse large B cell lymphomas (DLBCL. We have studied the influence on BLIMP1β expression of the Epstein-Barr virus (EBV, a human herpesvirus that is implicated in the pathogenesis of several GC-derived lymphomas, including a subset of DLBCL and Hodgkin’s lymphoma (HL. We show that BLIMP1β expression is increased following the EBV infection of normal human tonsillar GC B cells. We also show that this change in expression is accompanied by hypomethylation of the BLIMP1β-specific promoter. Furthermore, we confirmed previous reports that the BLIMP1β promoter is hypomethylated in DLBCL cell lines and show for the first time that BLIMP1β is hypomethylated in the Hodgkin/Reed-Sternberg (HRS cells of HL. Our results provide evidence in support of a role for BLIMP1β in the pathogenesis of EBV-associated B cell lymphomas.

  8. Dielectric barrier discharge plasma pretreatment on hydrolysis of microcrystalline cellulose

    Science.gov (United States)

    Huang, Fangmin; Long, Zhouyang; Liu, Sa; Qin, Zhenglong

    2017-04-01

    Dielectric barrier discharge (DBD) plasma was used as a pretreatment method for downstream hydrolysis of microcrystalline cellulose (MCC). The degree of polymerization (DP) of MCC decreased after it was pretreated by DBD plasma under a carrier gas of air/argon. The effectiveness of depolymerization was found to be influenced by the crystallinity of MCC when under the pretreatment of DBD plasma. With the addition of tert-butyl alcohol in the treated MCC water suspension solution, depolymerization effectiveness of MCC was inhibited. When MCC was pretreated by DBD plasma for 30 min, the total reducing sugar concentration (TRSC) and liquefaction yield (LY) of pretreated-MCC (PMCC) increased by 82.98% and 34.18% respectively compared with those for raw MCC.

  9. Pretreatment of Biomass by Aqueous Ammonia for Bioethanol Production

    Science.gov (United States)

    Kim, Tae Hyun; Gupta, Rajesh; Lee, Y. Y.

    The methods of pretreatment of lignocellulosic biomass using aqueous ammonia are described. The main effect of ammonia treatment of biomass is delignification without significantly affecting the carbohydrate contents. It is a very effective pretreatment method especially for substrates that have low lignin contents such as agricultural residues and herbaceous feedstock. The ammonia-based pretreatment is well suited for simultaneous saccharification and co-fermentation (SSCF) because the treated biomass retains cellulose as well as hemicellulose. It has been demonstrated that overall ethanol yield above 75% of the theoretical maximum on the basis of total carbohydrate is achievable from corn stover pretreated with aqueous ammonia by way of SSCF. There are two different types of pretreatment methods based on aqueous ammonia: (1) high severity, low contact time process (ammonia recycle percolation; ARP), (2) low severity, high treatment time process (soaking in aqueous ammonia; SAA). Both of these methods are described and discussed for their features and effectiveness.

  10. Effect of 5-fluorodeoxyuridine on DNA replication in ultraviolet-irradiated HeLa cells

    Energy Technology Data Exchange (ETDEWEB)

    Brozmanova, J.; Masek, F.; Synzynys, B.I.; Saenko, A.S.

    1985-11-05

    In HeLa cells precultivated for 6 hours with 5-fluorodeoxyuridine (FUdR) and for 18 hours in FUdR-free medium, DNA synthesis was much more resistant to UV irradiation than that of untreated cells. DNA synthesized in FUdR-pretreated and UV irradiated cells represents a semiconservative DNA replication and shows more rapid shift of the pulse-labelled chased DNA to high molecular weight. This DNA synthesis is not induced by synchronization of the cell cycle. It is assumed that either the changes of chromatine structure, or an enhanced level of some enzymes might be involved in the replication of the damaged template. (author).

  11. Niacinamide pretreatment reduces microvesicle formation in hairless guinea pigs cutaneously exposed to sulfur mustard. (Reannouncement with new availability information)

    Energy Technology Data Exchange (ETDEWEB)

    Yourick, J.J.; Clark, C.R.; Mitcheltree, L.W.

    1991-12-31

    It has been proposed that sulfur mustard (HD) may indirectly activate poly(ADP-ribose) polymerase (PADPRP) by alkylating cellular DNA (Papirmeister et al., 1985). Activation of PADPRP results in the depletion of cellular NAD+ which initiates a series of biochemical processes that have been proposed to culminate in blister formation. Preventing PADPRP activation and NAD+ depletion should inhibit blister formation. Niacinamide is both an inhibitor of PADPRP and a precursor for NAD+ synthesis. The present study was undertaken to determine whether niacinamide can protect against HD-induced microvesication in cutaneously exposed hairless guinea pigs. Each site was exposed to HD for 8 min by means of a vapor cup. Niacinamide (750 mg/kg, ip) given as a 30-min pretreatment inhibited microvesicle formation by 50% after HD application. However, niacinamide given 2 hr after HD application did not reduce microvesicle formation. There was no benefit when niacinamide was given as both a pretreatment and treatment when compared to niacinamide given only as a pretreatment. The reduction in microvesication 24 hr after HD did not correlate with skin NAD+ content. Niacinamide did not reduce the degree of erythema or edema. Ballooning degeneration of basal epidermal cells was present in some niacinamide pretreated HD exposure sites.

  12. DNA Investigations.

    Science.gov (United States)

    Mayo, Ellen S.; Bertino, Anthony J.

    1991-01-01

    Presents a simulation activity that allow students to work through the exercise of DNA profiling and to grapple with some analytical and ethical questions involving a couple arranging with a surrogate mother to have a baby. Can be used to teach the principles of restriction enzyme digestion, gel electrophoresis, and probe hybridization. (MDH)

  13. Modification of gelatin-DNA interaction for optimised DNA extraction from gelatin and gelatin capsule.

    Science.gov (United States)

    Mohamad, Nurhidayatul Asma; Mustafa, Shuhaimi; El Sheikha, Aly Farag; Khairil Mokhtar, Nur Fadhilah; Ismail, Amin; Ali, Md Eaqub

    2016-05-01

    Poor quality and quantity of DNA extracted from gelatin and gelatin capsules often causes failure in the determination of animal species using PCR. Gelatin, which is mainly derived from porcine and bovine, has been a matter of concern among customers in order to fulfill religious obligation and safety precaution against several transmissible infectious diseases associated with bovine species. Thus, optimised DNA extraction from gelatin is very important for successful real-time PCR detection of gelatin species. In this work, the DNA extraction method was optimised in terms of lysis incubation period and inclusion of pre-treatment pH modification of samples. The yield of DNA extracted from porcine gelatin was significantly increased when the pH of the samples was adjusted to pH 8.5 prior to DNA precipitation with isopropanol. The optimal pH for DNA precipitation from bovine gelatin solution was then determined at the original pH range of solution: pH 7.6 to 8. A DNA fragment of approximately 300 base pairs was available for PCR amplification. DNA extracted from gelatin and commercially available capsules has been successfully utilised for species detection using real-time PCR assay. However, significant adulterations of porcine and bovine in pure gelatin and capsules have been detected, which require further analytical techniques for validation. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  14. Low temperature lignocellulose pretreatment: effects and interactions of pretreatment pH are critical for maximizing enzymatic monosaccharide yields from wheat straw

    DEFF Research Database (Denmark)

    Pedersen, Mads; Johansen, Katja S.; Meyer, Anne S.

    2011-01-01

    Background: The recent development of improved enzymes and pentose-using yeast for cellulosic ethanol processes calls for new attention to the lignocellulose pretreatment step. This study assessed the influence of pretreatment pH, temperature, and time, and their interactions on the enzymatic...... alkaline pretreatments. Alkaline pretreatments also solubilized most of the lignin. Conclusions: Pretreatment pH exerted significant effects and factor interactions on the enzymatic glucose and xylose releases. Quite extreme pH values were necessary with mild thermal pretreatment strategies (T...... glucose and xylose yields from mildly pretreated wheat straw in multivariate experimental designs of acid and alkaline pretreatments. Results: The pretreatment pH was the most significant factor affecting both the enzymatic glucose and xylose yields after mild thermal pretreatments at maximum 140 degrees...

  15. Possible involvement of ROS generation in vorinostat pretreatment induced enhancement of the antibacterial activity of ciprofloxacin

    Directory of Open Access Journals (Sweden)

    Masadeh MM

    2017-10-01

    Full Text Available Majed M Masadeh,1 Karem H Alzoubi,2 Sayer I Al-azzam,2 Ahlam M Al-buhairan3 1Department of Pharmaceutical Technology, 2Department of Clinical Pharmacy, Jordan University of Science and Technology, Irbid, Jordan; 3Department of Clinical Laboratory Sciences, King Saud University, Riyadh, Saudi Arabia Abstract: The mechanism underlying ciprofloxacin action involves interference with transcription and replication of bacterial DNA and, thus, the induction of double-strand breaks in DNA. It also involves elevated oxidative stress, which might contribute to bacterial cell death. Vorinostat was shown to induce oxidative DNA damage. The current work investigated a possible interactive effect of vorinostat on ciprofloxacin-induced cytotoxicity against a number of reference bacteria. Standard bacterial strains were Escherichia coli ATCC 35218, Staphylococcus aureus ATCC29213, Pseudomonas aeruginosa ATCC 9027, Staphylococcus epidermidis ATCC 12228, Acinetobacter baumannii ATCC 17978, Proteus mirabilis ATCC 12459, Klebsiella pneumoniae ATCC 13883, methicillin-resistant Staphylococcus aureus (MRSA (ATCC 43300, and Streptococcus pneumoniae (ATCC 25923. The antibacterial activity of ciprofloxacin, with or without pretreatment of bacterial cells by vorinostat, was examined using the disc diffusion procedure and determination of the minimum inhibitory concentration (MIC and zones of inhibition of bacterial growth. All tested bacterial strains showed sensitivity to ciprofloxacin. When pretreated with vorinostat, significantly larger zones of inhibition and smaller MIC values were observed in all bacterial strains compared to those treated with ciprofloxacin alone. In correlation, generation of reactive oxygen species (ROS induced by the antibacterial action of ciprofloxacin was enhanced by treatment of bacterial cells with vorinostat. Results showed the possible agonistic properties of vorinostat when used together with ciprofloxacin. This could be related to the

  16. Lime pretreatment and fermentation of enzymatically hydrolyzed sugarcane bagasse.

    Science.gov (United States)

    Rabelo, Sarita C; Maciel Filho, Rubens; Costa, Aline C

    2013-03-01

    Sugarcane bagasse was subjected to lime (calcium hydroxide) pretreatment and enzymatic hydrolysis for second-generation ethanol production. A central composite factorial design was performed to determine the best combination of pretreatment time, temperature, and lime loading, as well as to evaluate the influence of enzymatic loadings on hydrolysis conversion. The influence of increasing solids loading in the pretreatment and enzymatic hydrolysis stages was also determined. The hydrolysate was fermented using Saccharomyces cerevisiae in batch and continuous mode. In the continuous fermentation, the hydrolysates were concentrated with molasses. Lime pretreatment significantly increased the enzymatic digestibility of sugarcane bagasse without the need for prior particle size reduction. In the optimal pretreatment conditions (90 h, 90 °C, 0.47 glime/g bagasse) and industrially realistic conditions of hydrolysis (12.7 FPU/g of cellulase and 7.3 CBU/g of β-glucosidase), 139.6 kglignin/ton raw bagasse and 126.0 kg hemicellulose in the pretreatment liquor per ton raw bagasse were obtained. The hydrolysate from lime pretreated sugarcane bagasse presented low amounts of inhibitors, leading to ethanol yield of 164.1 kgethanol/ton raw bagasse.

  17. Enhanced enzymatic conversion with freeze pretreatment of rice straw

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Ken-Lin; Thitikorn-amorn, Jitladda; Ou, Bay-Ming; Chen, Shan-He; Huang, Po-Jung [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Hsieh, Jung-Feng [Department of Food Science, Fu Jen Catholic University, Xin Zhuang, Taipei 242 (China); Ratanakhanokchai, Khanok [School of Bioresources and Technology, King Mongkut' s University of Technology Thonburi, Bangkok 10150 (Thailand); Chen, Shui-Tein [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 106 (China)

    2011-01-15

    Production of bioethanol by the conversion of lignocellulosic waste has attracted much interest in recent years, because of its low cost and great potential availability. The pretreatment process is important for increasing the enzymatic digestibility of lignocellulosic materials. Enzymatic conversion with freeze pretreatment of rice straw was evaluated in this study. The freeze pretreatment was found to significantly increase the enzyme digestibility of rice straw from 48% to 84%. According to the results, enzymatic hydrolysis of unpretreated rice straw with 150 U cellulase and 100 U xylanase for 48 h yielded 226.77 g kg{sup -1} and 93.84 g kg{sup -1} substrate-reducing sugars respectively. However, the reducing sugar yields from freeze pretreatment under the same conditions were 417.27 g kg{sup -1} and 138.77 g kg{sup -1} substrate, respectively. In addition, hydrolyzates analysis showed that the highest glucose yield obtained during the enzymatic hydrolysis step in the present study was 371.91 g kg{sup -1} of dry rice straw, following pretreatment. Therefore, the enhanced enzymatic conversion with freeze pretreatment of rice straw was observed in this study. This indicated that freeze pretreatment was highly effective for enzymatic hydrolysis and low environmental impact. (author)

  18. Understanding the impact of ionic liquid pretreatment on eucalyptus

    Energy Technology Data Exchange (ETDEWEB)

    Centikol, Ozgul [Joint Bioenergy Institute; Dibble, Dean [Joint Bioenergy Institute; Cheng, Gang [Joint Bioenergy Institute; Kent, Michael S [ORNL; Knierim, Manfred [Joint Bioenergy Institute; Melnichenko, Yuri B [ORNL

    2010-01-01

    The development of cost-competitive biofuels necessitates the realization of advanced biomass pretreatment technologies. Ionic liquids provide a basis for one of the most promising pretreatment technologies and are known to allow effective processing of cellulose and some biomass species. Here, we demonstrate that the ionic liquid 1-ethyl-3-methyl imidazolium acetate, [C2mim][OAc], induces structural changes at the molecular level in the cell wall of Eucalyptus globulus. Deacetylation of xylan, acetylation of the lignin units, selective removal of guaiacyl units (increasing the syringyl:guaiacyl ratio) and decreased {beta}-ether content were the most prominent changes observed. Scanning electron microscopy images of the plant cell wall sections reveal extensive swelling during [C2mim][OAc] pretreatment. X-ray diffraction measurements indicate a change in cellulose crystal structure from cellulose I to cellulose II after [C2mim][OAc] pretreatment. Enzymatic saccharification of the pretreated material produced increased sugar yields and improved hydrolysis kinetics after [C2mim][OAc] pretreatment. These results provide new insight into the mechanism of ionic liquid pretreatment and reaffirm that this approach may be promising for the production of cellulosic biofuels from woody biomass.

  19. Effects of Pretreatment Methods on Electrodes and SOFC Performance

    Directory of Open Access Journals (Sweden)

    Guo-Bin Jung

    2014-06-01

    Full Text Available Commercially available tapes (anode, electrolyte and paste (cathode were choosen to prepare anode-supported cells for solid oxide fuel cell applications. For both anode-supported cells or electrolyte-supported cells, the anode needs pretreatment to reduce NiO/YSZ to Ni/YSZ to increase its conductivity as well as its catalytic characteristics. In this study, the effects of different pretreatments (open-circuit, closed-circuit on cathode and anodes as well as SOFC performance are investigated. To investigate the influence of closed-circuit pretreatment on the NiO/YSZ anode alone, a Pt cathode is utilized as reference for comparison with the LSM cathode. The characterization of the electrical resistance, AC impedance, and SOFC performance of the resulting electrodes and/or anode-supported cell were carried out. It’s found that the influence of open-circuit pretreatment on the LSM cathode is limited. However, the influence of closed-circuit pretreatment on both the LSM cathode and NiO/YSZ anode and the resulting SOFC performance is profound. The effect of closed-circuit pretreatment on the NiO/YSZ anode is attributed to its change of electronic/pore structure as well as catalytic characteristics. With closed-circuit pretreatment, the SOFC performance improved greatly from the change of LSM cathode (and Pt reference compared to the Ni/YSZ anode.

  20. Dilute alkali pretreatment of softwood pine: A biorefinery approach.

    Science.gov (United States)

    Safari, Ali; Karimi, Keikhosro; Shafiei, Marzieh

    2017-06-01

    Dilute alkali pretreatment was performed on softwood pine to maximize ethanol and biogas production via a biorefinery approach. Alkali pretreatments were performed with 0-2% w/v NaOH at 100-180°C for 1-5h. The liquid fraction of the pretreated substrates was subjected to anaerobic digestion. The solid fraction of the pretreatment was used for separate enzymatic hydrolysis and fermentation. High ethanol yields of 76.9‒78.0% were achieved by pretreatment with 2% (w/v) NaOH at 180°C. The highest biogas yield of 244mL/g volatile solid (at 25°C, 1bar) was achieved by the pretreatment with 1% (w/v) NaOH at 180°C. The highest gasoline equivalent (sum of ethanol and methane) of 197L per ton of pinewood and the lowest ethanol manufacturing cost of 0.75€/L was obtained after pretreatment with 1% NaOH at 180°C for 5h. The manufacturing cost of ethanol from untreated wood was 4.12€/L. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. 40 CFR 439.37 - Pretreatment standards for new sources (PSNS).

    Science.gov (United States)

    2010-07-01

    ...) EFFLUENT GUIDELINES AND STANDARDS PHARMACEUTICAL MANUFACTURING POINT SOURCE CATEGORY Chemical Synthesis... achieve the pretreatment standard for ammonia (as N). (b) The pretreatment standards for cyanide are as...

  2. Effects of different pretreatment methods on fermentation types and dominant bacteria for hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Ren, Nan-Qi; Guo, Wan-Qian; Liu, Bing-Feng; Wang, Xing-Zu; Ding, Jie; Chen, Zhao-Bo [State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090, Heilongjiang (China); Wang, Xiang-Jing; Xiang, Wen-Sheng [Research Center of Life Science and Biotechnology, Northeast Agricultural University, Harbin 150030 (China)

    2008-08-15

    In order to enrich hydrogen producing bacteria and to establish high-efficient communities of the mixed microbial cultures, inoculum needs to be pretreated before the cultivation. Four pretreatment methods including heat-shock pretreatment, acid pretreatment, alkaline pretreatment and repeated-aeration pretreatment were performed on the seed sludge which was collected from a secondary settling tank of a municipal wastewater treatment plant. In contrast to the control test without any pretreatment, the heat-shock pretreatment, acid pretreatment and repeated-aeration pretreatment completely suppressed the methanogenic activity of the seed sludge, but the alkaline pretreatment did not. Employing different pretreatment methods resulted in the change in fermentation types as butyric-acid type fermentation was achieved by the heat-shock and alkaline pretreatments, mixed-acid type fermentation was achieved by acid pretreatment and the control, and ethanol-type fermentation was observed by repeated-aeration pretreatment. Denaturing gradient gel electrophoresis (DGGE) profiles revealed that pretreatment method substantially affected the species composition of microbial communities. The highest hydrogen yield of 1.96 mol/mol-glucose was observed with the repeated-aeration pretreatment method, while the lowest was obtained as the seed sludge was acidified. It is concluded that the pretreatment methods led to the difference in the initial microbial communities which might be directly responsible for different fermentation types and hydrogen yields. (author)

  3. TGF-β1 accelerates the DNA damage response in epithelial cells via Smad signaling

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jeeyong; Kim, Mi-Ra; Kim, Hyun-Ji; An, You Sun; Yi, Jae Youn, E-mail: yjy_71@kcch.re.kr

    2016-08-05

    The evidence suggests that transforming growth factor-beta (TGF-β) regulates the DNA-damage response (DDR) upon irradiation, and we previously reported that TGF-β1 induced DNA ligase IV (Lig4) expression and enhanced the nonhomologous end-joining repair pathway in irradiated cells. In the present study, we investigated the effects of TGF-β1 on the irradiation-induced DDRs of A431 and HaCaT cells. Cells were pretreated with or without TGF-β1 and irradiated. At 30 min post-irradiation, DDRs were detected by immunoblotting of phospho-ATM, phospho-Chk2, and the presence of histone foci (γH2AX). The levels of all three factors were similar right after irradiation regardless of TGF-β1 pretreatment. However, they soon thereafter exhibited downregulation in TGF-β1-pretreated cells, indicating the acceleration of the DDR. Treatment with a TGF-β type I receptor inhibitor (SB431542) or transfections with siRNAs against Smad2/3 or DNA ligase IV (Lig4) reversed this acceleration of the DDR. Furthermore, the frequency of irradiation-induced apoptosis was decreased by TGF-β1 pretreatment in vivo, but this effect was abrogated by SB431542. These results collectively suggest that TGF-β1 could enhance cell survival by accelerating the DDR via Smad signaling and Lig4 expression. -- Highlights: •TGF-β1 pretreatment accelerates γ-radiation-induced DNA damage response. •TGF-β1-accelerated DNA damage response is dependent on Smad signaling and DNA Ligase IV. •TGF-β1 pretreatment protects epithelial cells from γ-radiation in vivo.

  4. Influence of ultrasound pretreatment on wood physiochemical structure.

    Science.gov (United States)

    He, Zhengbin; Wang, Zhenyu; Zhao, Zijian; Yi, Songlin; Mu, Jun; Wang, Xiaoxu

    2017-01-01

    As an initial step to increase the use of renewable biomass resources, this study was aimed at investigating the effects of ultrasound pretreatment on structural changes of wood. Samples were pretreated by ultrasound with the power of 300W and frequency of 28kHz in aqueous soda solution, aqueous acetic acid, or distilled water, then pretreated and control samples were characterized via X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA). The results shown that ultrasound pretreatment is indeed effective in modifying the physiochemical structure of eucalyptus wood; the pretreatment decreased the quantity of alkali metals (e.g., potassium, calcium and magnesium) in the resulting material. Compared to the control group, the residual char content of samples pretreated in aqueous soda solution increased by 10.08%-20.12% and the reaction temperature decreased from 361°C to 341°C, however, in samples pretreated by ultrasound in acetic solution or distilled water, the residual char content decreased by 12.40%-21.45% and there were no significant differences in reactivity apart from a slightly higher maximum reaction rate. Ultrasound pretreatment increased the samples' crystallinity up to 35.5% and successfully removed cellulose, hemicellulose, and lignin from the samples; the pretreatment also increased the exposure of the sample to the treatment solutions, broke down sample pits, and generated collapses and microchannels on sample pits, and removed attachments in the samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Investigation of Pleurotus ostreatus pretreatment on switchgrass for ethanol production

    Science.gov (United States)

    Slavens, Shelyn Gehle

    Fungal pretreatment using the white-rot fungus Pleurotus ostreatus on switchgrass for ethanol production was studied. In a small-scale storage study, small switchgrass bales were inoculated with fungal spawn and automatically watered to maintain moisture. Sampled at 25, 53, and 81 d, the switchgrass composition was determined and liquid hot water (LHW) pretreatment was conducted. Fungal pretreatment significantly decreased the xylan and lignin content; glucan was not significantly affected by fungal loading. The glucan, xylan, and lignin contents significantly decreased with increased fungal pretreatment time. The effects of the fungal pretreatment were not highly evident after the LHW pretreatment, showing only changes based on sampling time. Although other biological activity within the bales increased cellulose degradation, the fungal pretreatment successfully reduced the switchgrass lignin and hemicellulose contents. In a laboratory-scale nutrient supplementation study, copper, manganese, glucose, or water was added to switchgrass to induce production of ligninolytic enzymes by P. ostreatus. After 40 d, ligninolytic enzyme activities and biomass composition were determined and simultaneous saccharification and fermentation (SSF) was conducted to determine ethanol yield. Laccase activity was similar for all supplements and manganese peroxidase (MnP) activity was significantly less in copper-treated samples than in the other fungal-inoculated samples. The fungal pretreatment reduced glucan, xylan, and lignin content, while increasing extractable sugars content. The lowest lignin contents occurred in the water-fungal treated samples and produced the greatest ethanol yields. The greatest lignin contents occurred in the copper-fungal treated samples and produced the lowest ethanol yields. Manganese-fungal and glucose-fungal treated samples had similar, intermediate lignin contents and produced similar, intermediate ethanol yields. Ethanol yields from switchgrass

  6. DNA Chip

    Indian Academy of Sciences (India)

    Imagine a world without identity cards; no I-cards for the college or office or bank account or anything! All you are carrying is a small (say, 2cm x 2cm) 'DNA-chip', which has the whole of your genetic profile on it. Your identity cannot get more authentic than that. Imagine a world where marriages are not decided by matching ...

  7. DNA repair

    International Nuclear Information System (INIS)

    Setlow, R.

    1978-01-01

    Some topics discussed are as follows: difficulty in extrapolating data from E. coli to mammalian systems; mutations caused by UV-induced changes in DNA; mutants deficient in excision repair; other postreplication mechanisms; kinds of excision repair systems; detection of repair by biochemical or biophysical means; human mutants deficient in repair; mutagenic effects of UV on XP cells; and detection of UV-repair defects among XP individuals

  8. High sensitivity but normal DNA-repair activity after UV irradiation in Epstein-Barr virus-transformed lymphoblastoid cell lines from Chediak-Higashi syndrome

    International Nuclear Information System (INIS)

    Tanaka, H.; Orii, T.

    1980-01-01

    We established lymphoblastoid cell lines from 2 children with Chediak-Higashi syndrome (CHS), 2 xeroderma pigmentosum (XP) patients and control donors after transformation of peripheral lymphocytes by Epstein-Barr virus (EBV). We used these lymphoblastoid cell lines to investigate repair activity after ultraviolet irradiation. Cell survival of both CHS lymphoblastoid cell lines after irradiation by UV and treatment by 4-nitroquinoline 1-oxide (4NQO) fell between those of the XP and control cells lines. Unscheduled DNA synthesis of CHS cells after UV irradiation occured at rates similar to those of control cells. (orig.)

  9. Stool DNA Test

    Science.gov (United States)

    ... The stool DNA test is a noninvasive laboratory test that identifies DNA changes in the cells of a stool sample. ... the presence of cancer. If a stool DNA test detects abnormal DNA, additional testing may be used to investigate the ...

  10. PROJECT W-551 INTERIM PRETREATMENT SYSTEM PRECONCEPTUAL CANDIDATE TECHNOLOGY DESCRIPTIONS

    Energy Technology Data Exchange (ETDEWEB)

    MAY TH

    2008-08-12

    The Office of River Protection (ORP) has authorized a study to recommend and select options for interim pretreatment of tank waste and support Waste Treatment Plant (WTP) low activity waste (LAW) operations prior to startup of all the WTP facilities. The Interim Pretreatment System (IPS) is to be a moderately sized system which separates entrained solids and 137Cs from tank waste for an interim time period while WTP high level waste vitrification and pretreatment facilities are completed. This study's objective is to prepare pre-conceptual technology descriptions that expand the technical detail for selected solid and cesium separation technologies. This revision includes information on additional feed tanks.

  11. Evaluation of wet oxidation pretreatment for enzymatic hydrolysis of softwood

    DEFF Research Database (Denmark)

    Palonen, H.; Thomsen, A.B.; Tenkanen, M.

    2004-01-01

    , and the compositions of solid and liquid fractions were analyzed. The solid fraction after wet oxidation contained 58-64% cellulose, 2-16% hemicellulose, and 24-30% lignin. The pretreatment series gave information about the roles of lignin and hemicellulose in the enzymatic hydrolysis. The temperature...... of the pretreatment, the residual hemicellulose content of the substrate, and the type of the commercial cellulase preparation used were the most important factors affecting the enzymatic hydrolysis. The highest sugar yield in a 72-h hydrolysis, 79% of theoretical, was obtained using a pretreatment of 200degrees...

  12. Automated extraction of DNA from biological stains on fabric from crime cases. A comparison of a manual and three automated methods

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Hjort, Benjamin B; Hansen, Thomas N

    2013-01-01

    The presence of PCR inhibitors in extracted DNA may interfere with the subsequent quantification and short tandem repeat (STR) reactions used in forensic genetic DNA typing. DNA extraction from fabric for forensic genetic purposes may be challenging due to the occasional presence of PCR inhibitors...... that may be co-extracted with the DNA. Using 120 forensic trace evidence samples consisting of various types of fabric, we compared three automated DNA extraction methods based on magnetic beads (PrepFiler Express Forensic DNA Extraction Kit on an AutoMate Express, QIAsyphony DNA Investigator kit either...... with the sample pre-treatment recommended by Qiagen or an in-house optimized sample pre-treatment on a QIAsymphony SP) and one manual method (Chelex) with the aim of reducing the amount of PCR inhibitors in the DNA extracts and increasing the proportion of reportable STR-profiles. A total of 480 samples were...

  13. Application of SCGE assay, classical cytogenetics and FISH for studying in vitro californium-252 neutrons irradiations and BSH pretreatment on human lymphocytes

    International Nuclear Information System (INIS)

    Florjan, D.; Niedzwiedz, W.; Schneider, K.; Cebulska-Wasilewska, A.; Kreft, A.

    1999-01-01

    Recently, there has been observed a tendency to apply various energy neutrons for tumor therapy and particularly low energy neutrons from various sources, including californium-252 source, for cancer radiotherapy based on the neutron capture. In this study peripheral blood lymphocytes were used as a model for human cells and three methods were applied to assess the effectiveness of californium-252 neutrons irradiations in vitro in normal cells or pre-treated with compound enriched in B-10 ion. Human blood samples or isolated lymphocytes were irradiated with neutrons from isotopic 252 Cf source at the Faculty of Nuclear Physics and Technics at University of Mining and Metallurgy (both neutron source and samples were placed in polyethylene block). Chemical pretreatment with BSH (Na 2 10 B 12 H 11 SH) was done to introduce boron-10 ion into cells in order to check any enhancement effect due to the process of boron neutron capture. Comet assay was done to investigate the DNA damage. Classical cytogenetics was applied to assess the frequencies of unstable aberrations (dicentrics and rings). Fluorescence in situ hybridization (FISH) with probes for chromosomes 1, 4 (14.3% of whole genome) and pancentromeric probe was performed to evaluate the frequencies of stable aberrations. Linear (or close to linear) increase of DNA damage and aberration frequency was observed with dose of radiation both for lymphocytes untreated or pre-treated with BSH. Very little differences (statistically insignificant) due to radiation dose and BSH pretreatment were observed in the frequencies of SCEs detected in the second mitosis. There is no significant difference between boron pre-treated and not treated cells, and even slightly higher effects were observed in case of the highest dose without BSH pretreatment. The level of translocations observed is comparable with the frequencies of dicentrics and rings. (author)

  14. Wet oxidation pretreatment of rape straw for ethanol production

    DEFF Research Database (Denmark)

    Arvaniti, Efthalia; Bjerre, Anne Belinda; Schmidt, Jens Ejbye

    2012-01-01

    Simultaneous Saccharification and Fermentation (SSF). To reduce the water use and increase the energy efficiency in WO pretreatment features like recycling liquid (filtrate), presoaking of rape straw in water or recycled filtrate before WO, skip washing pretreated solids (filter cake) after WO, or use of whole...... slurry (Filter cake + filtrate) in SSF were also tested. Except ethanol yields, pretreatment methods were evaluated based on achieved glucose yields, amount of water used, recovery of cellulose, hemicellulose, and lignin.The highest ethanol yield obtained was 67% after fermenting the whole slurry...... produced by WO at 205 °C for 3 min with 12 bar of oxygen gas pressure and featured with presoaking in water. At these conditions after pre-treatment, cellulose and hemicellulose was recovered quantitatively (100%) together with 86% of the lignin. WO treatments of 2–3 min at 205–210 °C with 12 bar of oxygen...

  15. Effects of Pretreatments in Convective Dehydration of Rosehip (Rosa eglanteria

    Directory of Open Access Journals (Sweden)

    Alejandra Mabellini

    2012-04-01

    Full Text Available The aim of this work was to experimentally determine drying curves for thin layer and bed drying of rosehip fruits, with and without pretreatments, to reduce processing times as a function of drying air operating variables, to propose dehydration kinetics of fruits and to determine its kinetic parameters for further use within drying simulation software. Fruits were pre-treated both chemically and mechanically, which included dipping the fruits in NaOH and ethyl oleate solutions; and cutting or perforating the fruit cuticle, respectively. Simulation models were then adopted to fit the kinetics drying data considering fruit volume shrinkage. These simple models minimized the calculation time during the simulation of deep-bed driers. Results show that pre-treatments reduced processing times up to 57%, and evaluated models satisfactorily predicted the drying of rosehip fruit. Effective mass diffusion coefficients were up to 4-fold greater when fruit was submitted to mechanical pretreatments.

  16. Ensiling as pretreatment of grass for lignocellulosic biomass conversion

    DEFF Research Database (Denmark)

    Ambye-Jensen, Morten

    an efficient production of ethanol. Lastly, the conversion of xylan was extremely low in both grass and grass silage. Optimization of the enzymatic saccharification of grass was attempted through improvement of the hemicellulase content in the enzyme blend. However, neither additional xylanases (Cellic HTec2......Development of sound technologies of biomass conversion will be increasingly important for many years to come as planetary bounderies drive the development towards a biobased society. Pretreatment of lignocellulosic biomass is, in this regard, an essential technology. Current pretreatment methods...... method with low cost and low energy requirements, plus brings about multiple advantages with regards to agricultural management. However, the pretreatment effect of ensiling, and the overall effects for further conversion are limited. In this study, ensiling was evaluated as a method of pretreatment...

  17. Optimization of microwave pretreatment on wheat straw for ethanol production

    DEFF Research Database (Denmark)

    Xu, Jian; Chen, Hongzhang; Kádár, Zsófia

    2011-01-01

    An orthogonal design (L9(34)) was used to optimize the microwave pretreatment on wheat straw for ethanol production. The orthogonal analysis was done based on the results obtained from the nine pretreatments. The effect of four factors including the ratio of biomass to NaOH solution, pretreatment...... time, microwave power, and the concentration of NaOH solution with three different levels on the chemical composition, cellulose/hemicellulose recoveries and ethanol concentration was investigated. According to the orthogonal analysis, pretreatment with the ratio of biomass to liquid at 80 g kg−1......, the NaOH concentration of 10 kg m−3, the microwave power of 1000 W for 15 min was confirmed to be the optimal condition. The ethanol yield was 148.93 g kg−1 wheat straw at this condition, much higher than that from the untreated material which was only 26.78 g kg−1....

  18. Effect of ultrasonic pretreatment on emulsion polymerization of styrene.

    Science.gov (United States)

    Nagatomo, Daichi; Horie, Takafumi; Hongo, Chizuru; Ohmura, Naoto

    2016-07-01

    This study investigated the effect of pretreatment of ultrasonic irradiation on emulsion polymerization of styrene to propose a process intensification method which gives high conversion, high reaction rate, and high energy efficiency. The solution containing styrene monomer was irradiated by a horn mounted on the ultrasonic transducer with the diameter of 5mm diameter and the frequency of 28 kHz before starting polymerization. The pretreatment of ultrasound irradiation as short as 1 min drastically improved monomer dispersion and increased reaction rate even under the agitation condition with low rotational speed of impeller. Furthermore, the ultrasonic pretreatment resulted in higher monomer concentration in polymer particles and produced larger polymer particles than conventional polymerization without ultrasonic pretreatment. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Pretreatment of microalgae to improve biogas production: a review.

    Science.gov (United States)

    Passos, Fabiana; Uggetti, Enrica; Carrère, Hélène; Ferrer, Ivet

    2014-11-01

    Microalgae have been intensively studied as a source of biomass for replacing conventional fossil fuels in the last decade. The optimization of biomass production, harvesting and downstream processing is necessary for enabling its full-scale application. Regarding biofuels, biogas production is limited by the characteristics of microalgae, in particular the complex cell wall structure of most algae species. Therefore, pretreatment methods have been investigated for microalgae cell wall disruption and biomass solubilization before undergoing anaerobic digestion. This paper summarises the state of the art of different pretreatment techniques used for improving microalgae anaerobic biodegradability. Pretreatments were divided into 4 categories: (i) thermal; (ii) mechanical; (iii) chemical and (iv) biological methods. According to experimental results, all of them are effective at increasing biomass solubilization and methane yield, pretreatment effect being species dependent. Pilot-scale research is still missing and would help evaluating the feasibility of full-scale implementation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. 40 CFR 417.134 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitations. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  1. 40 CFR 417.74 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do. [39... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  2. 40 CFR 417.54 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do. [40... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  3. 40 CFR 408.206 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55787... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  4. 40 CFR 408.104 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  5. 40 CFR 408.226 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ...: Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  6. 40 CFR 408.144 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard pH No limitation. BOD5 Do. Oil and grease Do. [40 FR 6439, Feb. 11... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  7. 40 CFR 408.294 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  8. 40 CFR 408.316 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55798... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  9. 40 CFR 408.14 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do. [40 FR... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  10. 40 CFR 417.124 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  11. 40 CFR 408.94 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  12. 40 CFR 417.84 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  13. 40 CFR 408.236 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55790... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  14. 40 CFR 408.154 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55781... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  15. 40 CFR 408.34 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  16. 40 CFR 408.44 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... existing source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  17. 40 CFR 408.306 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55797... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  18. 40 CFR 408.334 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55800... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  19. 40 CFR 417.44 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do. [40... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  20. 40 CFR 408.284 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55795... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  1. 40 CFR 408.256 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ...: Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  2. 40 CFR 408.286 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55795, Dec. 1, 1975, as... introduces process wastewater pollutants into a publicly owned treatment works must comply with 40 CFR part...

  3. 40 CFR 408.24 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... existing source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  4. 40 CFR 408.156 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55781, Dec. 1, 1975, as... introduces process wastewater pollutants into a publicly owned treatment works must comply with 40 CFR part...

  5. 40 CFR 408.186 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ...: Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  6. 40 CFR 417.104 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  7. 40 CFR 408.246 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55791... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  8. 40 CFR 408.176 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55783... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  9. 40 CFR 417.114 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  10. 40 CFR 408.326 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55799... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  11. 40 CFR 408.166 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55781... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  12. 40 CFR 408.304 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  13. 40 CFR 408.336 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55800, Dec. 1, 1975, as... introduces process wastewater pollutants into a publicly owned treatment works must comply with 40 CFR part...

  14. 40 CFR 417.14 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  15. 40 CFR 408.296 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55796... subpart that introduces process wastewater pollutants into a publicly owned treatment works must comply...

  16. 40 CFR 408.276 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55794... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  17. 40 CFR 408.244 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  18. 40 CFR 408.54 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do... existing source subject to this subpart that introduces process wastewater pollutants into a publicly owned...

  19. 40 CFR 417.94 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    .... Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do... subject to this subpart that introduces process wastewater pollutants into a publicly owned treatment...

  20. 40 CFR 408.196 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55786... this subpart that introduces process wastewater pollutants into a publicly owned treatment works must...

  1. Pretreatment Characteristics of Waste Oak Wood by Ammonia Percolation

    Science.gov (United States)

    Kim, Jun-Seok; Kim, Hyunjoon; Lee, Jin-Suk; Lee, Joon-Pyo; Park, Soon-Chul

    A log of waste oak wood collected from a Korean mushroom farm has been tested for ammonia percolation pretreatment. The waste log has different physical characteristics from that of virgin oak wood. The density of the waste wood was 30% lower than that of virgin oak wood. However, there is little difference in the chemical compositions between the woods. Due to the difference in physical characteristics, the optimal pretreatment conditions were also quite different. While for waste oak the optimum temperature was determined to be 130°C, for virgin oak wood the optimum pretreatment was only achieved at 170°C. Presoaking for 12 h with ammonia solution before pretreatment was helpful to increase the delignification efficiency.

  2. Pretreatment of Miscanthus for hydrogen production by Thermotoga elfii

    NARCIS (Netherlands)

    Vrije, de T.; Haas, de G.G.; Tan, G.B.; Keijsers, E.R.P.; Claassen, P.A.M.

    2002-01-01

    Pretreatment methods for the production of fermentable substrates from Miscanthus, a lignocellulosic biomass, were investigated. Results demonstrated an inverse relationship between lignin content and the efficiency of enzymatic hydrolysis of polysaccharides. High delignification values were

  3. Carbohydrate degradation mechanisms and compounds from pretreated biomass

    DEFF Research Database (Denmark)

    Rasmussen, Helena

    The formation of inhibitors during pretreatment of lignocellulosic feedstocks is a persistent problem, and notably the compounds that retard enzymatic cellulose conversion represent an obstacle for achieving optimal enzymatic productivity and high glucose yields. Compounds with many chemical......, reduced the level of oligophenolic compounds with 73 % compared to the original pretreatment and 41 % compared to the control. When pretreatment severity was increased the amount of xylooligosaccharides decreased whereas the amount of oligophenolic compounds increased. No new degradation compounds were...... pretreated wheat straw after enzymatic treatment. It was found that formation of the oligophenolic degradation compounds were common across biomass sources as sugar cane bagasse and oil palm empty fruit bunches. These findings were in line with that the oligophenolic compounds arise from reactions involving...

  4. Pretreatment cardiometabolic status in youth with early-onset psychosis

    DEFF Research Database (Denmark)

    Jensen, Karsten G.; Correll, Christoph U.; Rudå, Ditte

    2017-01-01

    Objective: To describe pretreatment cardiometabolic constitution in children and adolescents with first-episode psychosis (FEP). Methods: Baseline cardiometabolic assessment was performed in youths aged 12-17 years with FEP entering the Tolerability and Efficacy of Antipsychotics (TEA) trial...

  5. Green liquor pretreatment for improving enzymatic hydrolysis of corn stover.

    Science.gov (United States)

    Gu, Feng; Yang, Linfeng; Jin, Yongcan; Han, Qiang; Chang, Hou-min; Jameel, Hasan; Phillips, Richard

    2012-11-01

    Green liquor consists of sodium carbonate and sodium sulfide and is readily available in any kraft mills. The green liquor pretreatment process for bioethanol production was developed for wood chips. This process uses only proven technology and equipment currently used in a kraft pulp mill and has several additional advantages such as high sugar recovery and concentration, no inhibitive substances produced, as compared to acid-based pretreatment methods. The liquor was used to pretreat corn stover for enhancing enzymatic hydrolysis in bioethanol production. Pulp yield of 70% with 45% lignin removal was achieved under optimized conditions (8% total titratable alkali, 40% sulfidity and 140°C). About 70% of the original polysaccharides were converted into fermentable sugars, using 20 FPU/g-pulp of enzyme in the subsequent enzymatic hydrolysis. The result indicates that green liquor is a feasible pretreatment to improve the enzymatic saccharification of corn stover for bioethanol production. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Pretreatment of bituminous coal for hydrogenation

    Energy Technology Data Exchange (ETDEWEB)

    Hupfer, H.

    1943-03-29

    Bituminous coal became plastic at certain temperature ranges and swelled, causing plugging of preheater tubes. With few exceptions, this problem was encountered only when hydrogenating bituminous coal. This swelling often appeared in the coking of coals, and for the majority of the cases it was found that efforts to reduce the baking ability of bituminous coal also reduced the swelling of coal paste in the preheater tubes. Three methods of pretreatment to reduce swelling were preheating, oxidation (anoxidation), and sulfur treatment. The preheating by hot nitrogen at 250 atm for 3 hours showed no improvement in swelling characteristics, but produced a greater asphalt content and a lower amount of gasification. Absorption of oxygen was shown to increase the viscosity of the coal paste, while preheating in a vacuum showed no effects. Oxidation (anoxidation) increased the splitting and gasification and gave higher asphalt content. The baking ability of Saar coal could be destroyed by addition of 2.5% sulfur and heating for 1/sup 1///sub 2/ hours at 110/sup 0/C, in a stream of H/sub 2/S. The volatile content dropped significantly when the same coal was heated to 200/sup 0/ to 250/sup 0/C. When neutralized Saar coal was dried under a stream of H/sub 2/S, only slight deterioration of the yield was noted with equal degree of gasification. It was concluded that a more or less distinct improvement would be observed from the absorption of sulfur, but that residue processing would have to be altered to account for the sulfur content.

  7. Pretreatment seizure semiology in childhood absence epilepsy.

    Science.gov (United States)

    Kessler, Sudha Kilaru; Shinnar, Shlomo; Cnaan, Avital; Dlugos, Dennis; Conry, Joan; Hirtz, Deborah G; Hu, Fengming; Liu, Chunyan; Mizrahi, Eli M; Moshé, Solomon L; Clark, Peggy; Glauser, Tracy A

    2017-08-15

    To determine seizure semiology in children with newly diagnosed childhood absence epilepsy and to evaluate associations with short-term treatment outcomes. For participants enrolled in a multicenter, randomized, double-blind, comparative-effectiveness trial, semiologic features of pretreatment seizures were analyzed as predictors of treatment outcome at the week 16 to 20 visit. Video of 1,932 electrographic absence seizures from 416 participants was evaluated. Median seizure duration was 10.2 seconds; median time between electrographic seizure onset and clinical manifestation onset was 1.5 seconds. For individual seizures and by participant, the most common semiology features were pause/stare (seizure 95.5%, participant 99.3%), motor automatisms (60.6%, 86.1%), and eye involvement (54.9%, 76.5%). The interrater agreement for motor automatisms and eye involvement was good (72%-84%). Variability of semiology features between seizures even within participants was high. Clustering analyses revealed 4 patterns (involving the presence/absence of eye involvement and motor automatisms superimposed on the nearly ubiquitous pause/stare). Most participants experienced more than one seizure cluster pattern. No individual semiologic feature was individually predictive of short-term outcome. Seizure freedom was half as likely in participants with one or more seizure having the pattern of eye involvement without motor automatisms than in participants without this pattern. Almost all absence seizures are characterized by a pause in activity or staring, but rarely is this the only feature. Semiologic features tend to cluster, resulting in identifiable absence seizure subtypes with significant intraparticipant seizure phenomenologic heterogeneity. One seizure subtype, pause/stare and eye involvement but no motor automatisms, is specifically associated with a worse treatment outcome. © 2017 American Academy of Neurology.

  8. DNA Microarrays

    Science.gov (United States)

    Nguyen, C.; Gidrol, X.

    Genomics has revolutionised biological and biomedical research. This revolution was predictable on the basis of its two driving forces: the ever increasing availability of genome sequences and the development of new technology able to exploit them. Up until now, technical limitations meant that molecular biology could only analyse one or two parameters per experiment, providing relatively little information compared with the great complexity of the systems under investigation. This gene by gene approach is inadequate to understand biological systems containing several thousand genes. It is essential to have an overall view of the DNA, RNA, and relevant proteins. A simple inventory of the genome is not sufficient to understand the functions of the genes, or indeed the way that cells and organisms work. For this purpose, functional studies based on whole genomes are needed. Among these new large-scale methods of molecular analysis, DNA microarrays provide a way of studying the genome and the transcriptome. The idea of integrating a large amount of data derived from a support with very small area has led biologists to call these chips, borrowing the term from the microelectronics industry. At the beginning of the 1990s, the development of DNA chips on nylon membranes [1, 2], then on glass [3] and silicon [4] supports, made it possible for the first time to carry out simultaneous measurements of the equilibrium concentration of all the messenger RNA (mRNA) or transcribed RNA in a cell. These microarrays offer a wide range of applications, in both fundamental and clinical research, providing a method for genome-wide characterisation of changes occurring within a cell or tissue, as for example in polymorphism studies, detection of mutations, and quantitative assays of gene copies. With regard to the transcriptome, it provides a way of characterising differentially expressed genes, profiling given biological states, and identifying regulatory channels.

  9. Pretreatment of Ferulic Acid Protects Human Dermal Fibroblasts against Ultraviolet A Irradiation

    Science.gov (United States)

    Hahn, Hyung Jin; Kim, Ki Bbeum; Bae, Seunghee; Choi, Byung Gon; An, Sungkwan

    2016-01-01

    Background Approximately 90%~99% of ultraviolet A (UVA) ray reaches the Earth's surface. The deeply penetrating UVA rays induce the formation of reactive oxygen species (ROS), which results in oxidative stress such as photoproducts, senescence, and cell death. Thus, UVA is considered a primary factor that promotes skin aging. Objective Researchers investigated whether pretreatment with ferulic acid protects human dermal fibroblasts (HDFs) against UVA-induced cell damages. Methods HDF proliferation was analyzed using the water-soluble tetrazolium salt assay. Cell cycle distribution and intracellular ROS levels were assessed by flow cytometric analysis. Senescence was evaluated using a senescence-associated β-galactosidase assay, while Gadd45α promoter activity was analyzed through a luciferase assay. The expression levels of superoxide dismutase 1 (SOD1), catalase (CAT), xeroderma pigmentosum complementation group A and C, matrix metalloproteinase 1 and 3, as well as p21 and p16 were measured using quantitative real-time polymerase chain reaction. Results Inhibition of proliferation and cell cycle arrest were detected in cells that were irradiated with UVA only. Pretreatment with ferulic acid significantly increased the proliferation and cell cycle progression in HDFs. Moreover, ferulic acid pretreatment produced antioxidant effects such as reduced DCF intensity, and affected SOD1 and CAT mRNA expression. These effects were also demonstrated in the analysis of cell senescence, promoter activity, expression of senescent markers, and DNA repair. Conclusion These results demonstrate that ferulic acid exerts protective effects on UVA-induced cell damages via anti-oxidant and stress-inducible cellular mechanisms in HDFs. PMID:27904274

  10. Pretreatment of Cottage Cheese to Enhance Biogas Production

    Directory of Open Access Journals (Sweden)

    Vidhya Prabhudessai

    2014-01-01

    Full Text Available This study evaluated the possibility of pretreating selected solid fraction of an anaerobic digester treating food waste to lower the hydraulic retention time and increase the methane production. The study investigated the effect of different pretreatments (thermal, chemical, thermochemical and enzymatic for enhanced methane production from cottage cheese. The most effective pretreatments were thermal and enzymatic. Highest solubilisation of COD was observed in thermal pretreatment, followed by thermochemical. In single enzyme systems, lipase at low concentration gave significantly higher methane yield than for the experiments without enzyme additions. The highest lipase dosages decreased methane yield from cottage cheese. However, in case of protease enzyme an increase in concentration of the enzyme showed higher methane yield. In the case of mixed enzyme systems, pretreatment at 1 : 2 ratio of lipase : protease showed higher methane production in comparison with 1 : 1 and 2 : 1 ratios. Methane production potentials for different pretreatments were as follows: thermal 357 mL/g VS, chemical 293 mL/g VS, and thermochemical 441 mL/g VS. The average methane yield from single enzyme systems was 335 mL/g VS for lipase and 328 mL/g VS for protease. Methane potentials for mixed enzyme ratios were 330, 360, and 339 mL/g VS for 1 : 1, 1 : 2, and 2 : 1 lipase : protease, respectively.

  11. Coagulation pretreatment of highly concentrated acrylonitrile wastewater from petrochemical plants.

    Science.gov (United States)

    Zheng, Dongju; Qin, Lin; Wang, Tao; Ren, Xiaojing; Zhang, Zhongguo; Li, Jiding

    2014-01-01

    Acrylonitrile (AN) wastewater is a heavily polluted and a likely hazardous liquid that is generated during the production of AN. Several chemical methods for the pretreatment of AN wastewater are available in laboratory scale. However, the harsh reaction conditions and high operational cost make these methods undesirable. Until now, four-effect evaporation is the only pretreatment method used for AN wastewater in industry despite its huge energy consumption and high cost. It is difficult to find an energy-saving pretreatment technique from the perspective of industrial application. In this study, a safe and low-cost coagulation technique was developed for the pretreatment of AN wastewater. Three types of inorganic coagulant and three types of polymer coagulant were investigated for the coagulation treatment of highly concentrated AN wastewater from petrochemical plants. The effects of coagulant type, dosage, and coagulation conditions on the pretreatment efficiency of AN wastewater were investigated. The results show that a combination of inorganic and polymer coagulants is effective for the pretreatment of AN wastewater.

  12. Influence of pretreatment of cotton yarns prior to biopolishing.

    Science.gov (United States)

    Ulson de Souza, A A; Ferreira, F C S; Guelli U Souza, S M A

    2013-04-02

    Cellulase is one of the enzymes most commonly used in the textile industry for the biopolishing process. The appropriate choice of pretreatment is a possible route to promoting enzymatic attack in situations in which this is not favored due to the effects of packing. In order to evaluate the influence of pretreatment the yarn was maintained in water for 24h before biopolishing to promote greater spacing between the chains. In the tensile testing the pretreated Combed 13/1 yarn showed a greater percentage reduction in the maximum breaking force following biopolishing, evidencing a stronger enzymatic attack. Also, the Combed 13/1 and OE 14/1 yarns without pretreatment had an approximately 22% reduction in the shrinkage and after pretreatment the Carded 13/1 yarn had the best shrinkage reduction values (18%). These data demonstrate that the introduction of the pretreatment promotes a change in the access of the enzyme to the fiber. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. New prospects in pretreatment of cotton fabrics using microwave heating.

    Science.gov (United States)

    Hashem, M; Taleb, M Abou; El-Shall, F N; Haggag, K

    2014-03-15

    As microwaves are known to give fast and rapid volume heating, the present study is undertaken to investigate the use of microwave heating for pretreatment cotton fabrics to reduce the pretreatment time, chemicals and water. The onset of the microwave heating technique on the physicochemical and performance properties of desized, scoured and bleached cotton fabric is elucidated and compared with those obtained on using conventional thermal heating. Combined one-step process for desizing, scouring and bleaching of cotton fabric under microwave heating was also investigated. The dual effect of adding urea, (as microwave absorber and hydrogen peroxide activator) has been exploiting to accelerate the pretreatment reaction of cotton fabric. DSC, FT-IR and SEM have been used to investigate the onset of microwave on the morphological and chemical change of cotton cellulose after pretreatment and bleaching under microwave heating. Results obtained show that, a complete fabric preparation was obtained in just 5 min on using microwave in pretreatments process and the fabric properties were comparable to those obtained in traditional pretreatment process which requires 2.5-3h for completion. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Effect of alkaline pretreatment on anaerobic digestion of solid wastes

    International Nuclear Information System (INIS)

    Lopez Torres, M.; Espinosa Llorens, Ma. del C.

    2008-01-01

    The introduction of the anaerobic digestion for the treatment of the organic fraction of municipal solid waste (OFMSW) is currently of special interest. The main difficulty in the treatment of this waste fraction is its biotransformation, due to the complexity of organic material. Therefore, the first step must be its physical, chemical and biological pretreatment for breaking complex molecules into simple monomers, to increase solubilization of organic material and improve the efficiency of the anaerobic treatment in the second step. This paper describes chemical pretreatment based on lime addition (Ca(OH) 2 ), in order to enhance chemical oxygen demand (COD) solubilization, followed by anaerobic digestion of the OFMSW. Laboratory-scale experiments were carried out in completely mixed reactors, 1 L capacity. Optimal conditions for COD solubilization in the first step of pretreatment were 62.0 mEq Ca(OH) 2 /L for 6.0 h. Under these conditions, 11.5% of the COD was solubilized. The anaerobic digestion efficiency of the OFMSW, with and without pretreatment, was evaluated. The highest methane yield under anaerobic digestion of the pretreated waste was 0.15 m 3 CH 4 /kg volatile solids (VS), 172.0% of the control. Under that condition the soluble COD and VS removal were 93.0% and 94.0%, respectively. The results have shown that chemical pretreatment with lime, followed by anaerobic digestion, provides the best results for stabilizing the OFMSW

  15. Dilute sulfuric acid pretreatment of sunflower stalks for sugar production.

    Science.gov (United States)

    Ruiz, Encarnación; Romero, Inmaculada; Moya, Manuel; Cara, Cristóbal; Vidal, Juan D; Castro, Eulogio

    2013-07-01

    In this work the pretreatment of sunflower stalks by dilute sulfuric acid is studied. Pretreatment temperature and the concentration of acid solution were selected as operation variables and modified according to a central rotatable composite experimental design. Based on previous studies pretreatment time was kept constant (5 min) while the variation range for temperature and acid concentration was centered at 175°C and 1.25% (w/v) respectively. Following pretreatment the insoluble solids were separated by filtration and further submitted to enzymatic hydrolysis, while liquid fractions were analyzed for sugars and inhibitors. Response surface methodology was applied to analyze results based on the combined severity of pretreatment experiments. Optimized results show that up to 33 g of glucose and xylose per 100g raw material (65% of the glucose and xylose present in the raw material) may be available for fermentation after pretreatment at 167°C and 1.3% sulfuric acid concentration. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Detection of Herpesviridae in whole blood by multiplex PCR DNA-based microarray analysis after hematopoietic stem cell transplantation.

    Science.gov (United States)

    Debaugnies, France; Busson, Laurent; Ferster, Alina; Lewalle, Philippe; Azzi, Nadira; Aoun, Mickael; Verhaegen, Godelieve; Mahadeb, Bhavna; de Marchin, Jérôme; Vandenberg, Olivier; Hallin, Marie

    2014-07-01

    Viral infections are important causes of morbidity and mortality in patients after hematopoietic stem cell transplantation. The monitoring by PCR of Herpesviridae loads in blood samples has become a critical part of posttransplant follow-up, representing mounting costs for the laboratory. In this study, we assessed the clinical performance of the multiplex PCR DNA microarray Clart Entherpex kit for detection of cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesvirus 6 (HHV-6) as a screening test for virological follow-up. Two hundred fifty-five blood samples from 16 transplanted patients, prospectively tested by routine PCR assays, were analyzed by microarray. Routine PCR detected single or multiple viruses in 42% and 10% of the samples, respectively. Microarray detected single or multiple viruses in 34% and 18% of the samples, respectively. Microarray results correlated well with CMV and EBV detections by routine PCR (kappa tests = 0.79 and 0.78, respectively), whereas a weak correlation was observed with HHV-6 (0.43). HHV-7 was also detected in 48 samples by microarray. In conclusion, the microarray is a reliable screening assay for a posttransplant virological follow-up to detect CMV and EBV infections in blood. However, positive samples must be subsequently confirmed and viral loads must be quantified by PCR assays. Limitations were identified regarding HHV-6 detection. Although it is promising, is easy to use as a first-line test, and allows a reduction in the cost of analysis without undue delay in the reporting of the final quantitative result to the clinician, some characteristics of this microarray should be improved, particularly regarding quality control and the targeted virus panel, such that it could then be used as a routine test. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  17. Techniques and practices for pretreatment of low and intermediate level solid and liquid radioactive wastes

    International Nuclear Information System (INIS)

    1987-01-01

    An overall waste management strategy generally includes several components: pretreatment, treatment, conditioning, transport and disposal. Benefits of pretreatment are improved safety, lower radiation exposures and significantly lower costs in subsequent waste management operations. This publication reviews current practices in the pretreatment of wastes in different countries and may assist the specialist in selection of appropriate pretreatment techniques

  18. Pretreatment of woody biomass for biofuel production: energy efficiency, technologies, and recalcitrance

    Science.gov (United States)

    J.Y. Zhu; Xuejun Pan; Ronald S. Jr. Zalesny

    2010-01-01

    This mini review discusses several key technical issues associated with cellulosic ethanol production from woody biomass: energy consumption for woody biomass pretreatment, pretreatment energy efficiency, woody biomass pretreatment technologies, and quantification of woody biomass recalcitrance. Both total sugar yield and pretreatment energy efficiency, defined as the...

  19. Woody biomass pretreatment for cellulosic ethanol production : technology and energy consumption evaluation

    Science.gov (United States)

    Junyong Zhu; X.J. Pan

    2010-01-01

    This review presents a comprehensive discussion of the key technical issues in woody biomass pretreatment: barriers to efficient cellulose saccharification, pretreatment energy consumption, in particular energy consumed for wood-size reduction, and criteria to evaluate the performance of a pretreatment. A post-chemical pretreatment size-reduction approach is proposed...

  20. BRCA1 Regulates IFI16 Mediated Nuclear Innate Sensing of Herpes Viral DNA and Subsequent Induction of the Innate Inflammasome and Interferon-β Responses.

    Directory of Open Access Journals (Sweden)

    Dipanjan Dutta

    2015-06-01

    Full Text Available The innate immune system pattern recognition receptors (PRR are the first line of host defenses recognizing the various pathogen- or danger-associated molecular patterns and eliciting defenses by regulating the production of pro-inflammatory cytokines such as IL-1β, IL-18 or interferon β (IFN-β. NOD-like receptors (NLRs and AIM2-like receptors (ALRs are cytoplasmic inflammasome sensors of foreign molecules, including DNA. IFI16, a sequence-independent nuclear innate sensor ALR, recognizes episomal dsDNA genomes of herpes viruses such as KSHV, EBV, and HSV-1 in the infected cell nuclei, forms an inflammasome complex with ASC and procaspase1, and relocates into the cytoplasm leading into Caspase-1 and IL-1β generation. IFI16 also induces IFN-β during HSV-1 infection via the cytoplasmic STING-TBK1-IRF3 pathway. Thus far, whether IFI16 recognizes foreign DNA directly or utilizes other host protein(s is unknown. Here, we demonstrate that BRCA1, a DNA damage repair sensor and transcription regulator, is in complex with IFI16 in the host cell nucleus, and their association increases in the presence of nuclear viral genomes during de novo KSHV, EBV and HSV-1 infection, and in latent KSHV or EBV infection, but not by DNA damage responses (DDR induced by bleomycin and vaccinia virus cytoplasmic dsDNA. BRCA1 is a constituent of the triggered IFI16-inflammasome and is translocated into the cytoplasm after genome recognition along with the IFI16-inflammasome. The absence of BRCA1 abrogated IFI16-viral genome association, inflammasome assembly, IFI16 cytoplasmic localization, and Caspase-1 and IL-1β production. The absence of BRCA1 also abolished the cytoplasmic IFI16-STING interaction, downstream IRF3 phosphorylation, nuclear translocation of pIRF3 and IFN-β production during de novo KSHV and HSV-1 infection. These findings highlight that BRCA1 plays a hitherto unidentified innate immunomodulatory role by facilitating nuclear foreign DNA sensing by

  1. DNA immunisation. New histochemical and morphometric data

    Directory of Open Access Journals (Sweden)

    D Ehirchiou

    2010-01-01

    Full Text Available Splenic germinal center reactions were measured during primary response to a plasmidic DNA intramuscular injection. Cardiotoxin-pretreated Balb/c mice were immunized with DNA plasmids encoding or not the SAG1 protein, a membrane antigen of Toxoplasma gondii. Specific anti-SAG1 antibodies were detected on days 16 and 36 after injection of coding plasmids. The results of ELISAs showed that the SAG1-specific antibodies are of the IgG2a class. Morphometric analyses were done on serial immunostained cryosections of spleen and draining or non-draining lymph nodes. This new approach made it possible to evaluate the chronological changes induced by DNA immunisation in the germinal centres (in number and in size. Significant increases in the number of germinal centres were measured in the spleen and only in draining lymph nodes after plasmid injection. the measured changes of the germinal centers appeared to result from the adjuvant stimulatory effect of the plasmidic DNA since both the coding and the noncoding plasmid DNA induced them. No measurable changes were recorded in the Tdependent zone of lymph organs.

  2. Biomass pretreatment affects Ustilago maydis in producing itaconic acid

    Directory of Open Access Journals (Sweden)

    Klement Tobias

    2012-04-01

    Full Text Available Abstract Background In the last years, the biotechnological production of platform chemicals for fuel components has become a major focus of interest. Although ligno-cellulosic material is considered as suitable feedstock, the almost inevitable pretreatment of this recalcitrant material may interfere with the subsequent fermentation steps. In this study, the fungus Ustilago maydis was used to produce itaconic acid as platform chemical for the synthesis of potential biofuels such as 3-methyltetrahydrofuran. No studies, however, have investigated how pretreatment of ligno-cellulosic biomass precisely influences the subsequent fermentation by U. maydis. Thus, this current study aims to first characterize U. maydis in shake flasks and then to evaluate the influence of three exemplary pretreatment methods on the cultivation and itaconic acid production of this fungus. Cellulose enzymatically hydrolysed in seawater and salt-assisted organic-acid catalysed cellulose were investigated as substrates. Lastly, hydrolysed hemicellulose from fractionated beech wood was applied as substrate. Results U. maydis was characterized on shake flask level regarding its itaconic acid production on glucose. Nitrogen limitation was shown to be a crucial condition for the production of itaconic acid. For itaconic acid concentrations above 25 g/L, a significant product inhibition was observed. Performing experiments that simulated influences of possible pretreatment methods, U. maydis was only slightly affected by high osmolarities up to 3.5 osmol/L as well as of 0.1 M oxalic acid. The production of itaconic acid was achieved on pretreated cellulose in seawater and on the hydrolysed hemicellulosic fraction of pretreated beech wood. Conclusion The fungus U. maydis is a promising producer of itaconic acid, since it grows as single cells (yeast-like in submerged cultivations and it is extremely robust in high osmotic media and real seawater. Moreover, U. maydis can grow on

  3. White-rot fungi pretreatment combined with alkaline/oxidative pretreatment to improve enzymatic saccharification of industrial hemp.

    Science.gov (United States)

    Xie, Chunliang; Gong, Wenbing; Yang, Qi; Zhu, Zuohua; Yan, Li; Hu, Zhenxiu; Peng, Yuande

    2017-11-01

    White-rot fungi combined with alkaline/oxidative (A/O) pretreatments of industrial hemp woody core were proposed to improve enzymatic saccharification. In this study, hemp woody core were treated with only white rot fungi, only A/O and combined with the two methods. The results showed that Pleurotus eryngii (P. eryngii) was the most effective fungus for pretreatment. Reducing sugars yield was 329mg/g with 30 Filter Paper Unit (FPU)/g cellulase loading when treated 21day. In the A/O groups, the results showed that when treated with 3% NaOH and 3% H 2 O 2 , the yield of reducing sugars was 288mg/g with 30FPU/g cellulase loading. After combination pretreatment with P. eryngii and A/O pretreatment, the reducing sugar yield from enzymatic hydrolysis of combined sample increased 1.10-1.29-fold than that of bio-treated or A/O pretreatment sample at the same conditions, suggesting that P. eryngii combined with A/O pretreatment was an effective method to improve enzyme hydrolysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Modeling of pretreatment condition of extrusion-pretreated prairie cordgrass and corn stover with poly (oxyethylen)20 sorbitan monolaurate.

    Science.gov (United States)

    Eckard, Anahita Dehkhoda; Muthukumarappan, Kasiviswanathan; Gibbons, William

    2012-05-01

    Extrusion processing has shown potential to be used as a pretreatment method for second-generation bioethanol production. Furthermore, surfactants have been shown to reduce enzyme deactivation and increase the efficiency of hydrolysis. Therefore, a sequential pretreatment technique was developed for corn stover (CS) and prairie cordgrass (PCG) in which a single screw extruder was used for the first pretreatment according to a previously optimized condition using 70-180 °C for feed, barrel, and die zones with 65-155 rpm screw speed. The second pretreatment was optimized in this study at 45-55 °C, 1-4 h, 0.15-0.6 g Tween 20/g glucan according to response surface methodology. Optimization of surfactant pretreatment facilitated the estimation of interaction and higher-order effects for major factors involved in surfactant treatment (temperature, time, surfactant loading). Using 8.6 FPU/g glucan cellulase, the optimum conditions found by fitting appropriate quadratic models to the data increased glucose and xylose yield by 27.5 and 33% for CS and by 21.5 and 27% for PCG, respectively. Tween 20 concentrations and pretreatment temperature were the most significant factors affecting sugar yield (p value <0.05). Studies of SDS concentration at and beyond critical micelle concentration (5.2-100 mM) demonstrated a decrease in sugar yield compared to control.

  5. Defining Essential Biodiversity Variables (EBVs) as a contribution to Essential Ocean Variables (EOVs): A Core Task of the Marine Biodiversity Observation Network (MBON) to Accelerate Integration of Biological Observations in the Global Ocean Observing System (GOOS)

    Science.gov (United States)

    Pearlman, J.; Muller-Karger, F. E.; Sousa Pinto, I.; Costello, M. J.; Duffy, J. E.; Appeltans, W.; Fischer, A. S.; Canonico, G.; Klein, E.; Obura, D.; Montes, E.; Miloslavich, P.; Howard, M.

    2017-12-01

    The Marine Biodiversity Observation Network (MBON) is a networking effort under the umbrella of the Group on Earth Observations Biodiversity Observation Network (GEO BON). The objective of the MBON is to link existing groups engaged in ocean observation and help define practical indices to deploy in an operational manner to track changes in the number of marine species, the abundance and biomass of marine organisms, the diverse interactions between organisms and the environment, and the variability and change of specific habitats of interest. MBON serves as the biodiversity arm of Blue Planet, the initiative of the Group on Earth Observations (GEO) for the benefit of society. The Global Ocean Observing System (GOOS) was established under the auspices of the Intergovernmental Oceanographic Commission (IOC) in 1991 to organize international ocean observing efforts. The mission of the GOOS is to support monitoring to improve the management of marine and coastal ecosystems and resources, and to enable scientific research. GOOS is engaged in a continuing, rigorous process of identifying Essential Ocean Variables (EOVs). MBON is working with GOOS and the Ocean Biogeographic Information System (OBIS, also under the IOC) to define Essential Biodiversity Variables (EBVs) as those Essential Ocean Variables (EOVs) that have explicit taxonomic records associated with them. For practical purposes, EBVs are a subset of the EOVs. The focus is to promote the integration of biological EOVs including EBVs into the existing and planned national and international ocean observing systems. The definition avoids a proliferation of 'essential' variables across multiple organizations. MBON will continue to advance practical and wide use of EBVs and related EOV. This is an effective way to contribute to several UN assessments (e.g., from IPBES, IPCC, and the World Ocean Assessment under the UN Regular Process), UN Sustainable Development Goals, and to address targets and goals defined under

  6. Tailoring Wet Explosion Process Parameters for the Pretreatment of Cocksfoot Grass for High Sugar Yields

    DEFF Research Database (Denmark)

    Njoku, Stephen Ikechukwu; Ahring, Birgitte Kiær; Uellendahl, Hinrich

    2013-01-01

    The pretreatment of lignocellulosic biomass is crucial for efficient subsequent enzymatic hydrolysis and ethanol fermentation. In this study, wet explosion (WEx) pretreatment was applied to cocksfoot grass and pretreatment conditions were tailored for maximizing the sugar yields using response su...... when applying less severe pretreatment conditions C (160 °C, 5 min, 0.2 % dilute sulfuric acid). Therefore, the choice of the most suitable pretreatment conditions is depending on the main target product, i.e., hexose or pentose sugars....

  7. Silane pre-treatments on copper and aluminium

    International Nuclear Information System (INIS)

    Deflorian, F.; Rossi, S.; Fedrizzi, L.

    2006-01-01

    A large part of aluminium products are coated with an organic layer in order to improve the corrosion resistance. Copper surfaces are also sometimes protected with an organic coating to improve the durability or the aesthetic properties. Examples of industrial applications are household appliances and heat exchanger components. For these applications it is not rare to have the industrial need to treat at the same time components made of aluminium and copper. In order to extend the service life of the organic coated copper a specific surface pre-treatment is often required. Nevertheless, probably because of the limited market of this application, no specific pre-treatments for copper are industrially developed, with the exception of cleaning procedures, but simply extensions of existing pre-treatments optimised for other metals (aluminium, zinc) are used. The application of silane pre-treatments as adhesion promoters for organic coated metals is remarkably increasing in the last decade, because silanes offer very good performance together with high environmental compatibility. The idea is therefore to try to develop a specific silane based pre-treatment for copper. The starting point is the existing silane products for aluminium, optimising the composition and the application conditions (concentration, temperature, pH of the bath, etc.) in order to develop a high performance copper alloy pre-treatment increasing the protective properties and the adhesion of a successively applied organic coating. Moreover these pre-treatments could be used for aluminium alloys too and therefore could be suggested for multi-metals components. The deposits were analysed using FTIR spectroscopy and optical and electron microscopic observations. A careful electrochemical characterisation, mainly by electrochemical impedance spectroscopy measurements (EIS) was carried out to highlight the presence of silane and to evaluate the performance of the different deposits. In order to study an

  8. Activation of glassy carbon electrodes by photocatalytic pretreatment

    Energy Technology Data Exchange (ETDEWEB)

    Dumanli, Onur [Department of Chemistry, Faculty of Science and Art, Ondokuz Mayis University, Kurupelit, 55139 Samsun (Turkey); Onar, A. Nur [Department of Chemistry, Faculty of Science and Art, Ondokuz Mayis University, Kurupelit, 55139 Samsun (Turkey)], E-mail: nonar@omu.edu.tr

    2009-11-01

    This paper describes a simple and rapid photocatalytic pretreatment procedure that removes contaminants from glassy carbon (GC) surfaces. The effectiveness of TiO{sub 2} mediated photocatalytic pretreatment procedure was compared to commonly used alumina polishing procedure. Cyclic voltammetric and chronocoulometric measurements were carried out to assess the changes in electrode reactivity by using four redox systems. Electrochemical measurements obtained on photocatalytically treated GC electrodes showed a more active surface relative to polished GC. In cyclic voltammograms of epinephrine, Fe(CN){sub 6}{sup 3-/4-} and ferrocene redox systems, higher oxidation and reduction currents were observed. The heterogeneous electron transfer rate constants (k{sup o}) were calculated for Fe(CN){sub 6}{sup 3-/4-} and ferrocene which were greater for photocatalytic pretreatment. Chronocoulometry was performed in order to find the amount of adsorbed methylene blue onto the electrode and was calculated as 0.34 pmol cm{sup -2} for photocatalytically pretreated GC. The proposed photocatalytic GC electrode cleansing and activating pretreatment procedure was more effective than classical alumina polishing.

  9. ALA Pretreatment Improves Waterlogging Tolerance of Fig Plants.

    Directory of Open Access Journals (Sweden)

    Yuyan An

    Full Text Available 5-aminolevulinic acid (ALA, a natural and environmentally friendly plant growth regulator, can improve plant tolerance to various environmental stresses. However, whether ALA can improve plant waterlogging tolerance is unknown. Here, we investigated the effects of ALA pretreatment on the waterlogging-induced damage of fig (Ficus carica Linn. plants, which often suffer from waterlogging stress. ALA pretreatment significantly alleviated stress-induced morphological damage, increased leaf relative water content (RWC, and reduced leaf superoxide anion ([Formula: see text] production rate and malonaldehyde (MDA content in fig leaves, indicating ALA mitigates waterlogging stress of fig plants. We further demonstrated that ALA pretreatment largely promoted leaf chlorophyll content, photosynthetic electron transfer ability, and photosynthetic performance index, indicating ALA significantly improves plant photosynthetic efficiency under waterlogging stress. Moreover, ALA pretreatment significantly increased activities of leaf superoxide dismutase (SOD and peroxidase (POD, root vigor, and activities of root alcohol dehydrogenase (ADH, and lactate dehydrogenase (LDH, indicating ALA also significantly improves antioxidant ability and root function of fig plants under waterlogging stress. Taken together, ALA pretreatment improves waterlogging tolerance of fig plants significantly, and the promoted root respiration, leaf photosynthesis, and antioxidant ability may contribute greatly to this improvement. Our data firstly shows that ALA can improve plant waterlogging tolerance.

  10. Autohydrolysis pretreatment assessment in ethanol production from agave bagasse.

    Science.gov (United States)

    Rios-González, Leopoldo J; Morales-Martínez, Thelma K; Rodríguez-Flores, María F; Rodríguez-De la Garza, José A; Castillo-Quiroz, David; Castro-Montoya, Agustín J; Martinez, Alfredo

    2017-10-01

    The aim of the present work was to assess the autohydrolysis pretreatment of Agave tequilana bagasse for ethanol production. The pretreatment was conducted using a one-liter high pressure Parr reactor under different severity factors (SF) at a 1:6w/v ratio (solid:liquid) and 200rpm. The solids obtained under the selected autohydrolysis conditions were subjected to enzymatic hydrolysis with a commercial cellulase cocktail, and the enzymatic hydrolysate was fermented using Saccharomyces cerevisiae. The results obtained from the pretreatment process showed that the glucan content in the pretreated solid was mostly preserved, and an increase in the digestibility was observed for the case with a SF of 4.13 (190°C, 30min). Enzymatic hydrolysis of the pretreated solids showed a yield of 74.3%, with a glucose concentration of 126g/L, resulting in 65.26g/L of ethanol after 10h of fermentation, which represent a 98.4% conversion according to the theoretical ethanol yield value. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Specific pretreatments reduce curing period of vanilla (Vanilla planifolia) beans.

    Science.gov (United States)

    Sreedhar, R V; Roohie, K; Venkatachalam, L; Narayan, M S; Bhagyalakshmi, N

    2007-04-18

    With the aiming of reducing the curing period, effects of pretreatments on flavor formation in vanilla beans during accelerated curing at 38 degrees C for 40 days were studied. Moisture loss, change in texture, levels of flavoring compounds, and activities of relevant enzymes were compared among various pretreatments as well as the commercial sample. Use of naphthalene acetic acid (NAA; 5 mg/L) or Ethrel (1%) with blanching pretreatment resulted in 3-fold higher vanillin on the 10th day. Other flavoring compounds-vanillic acid, p-hydroxybenzoic acid, and p-hydroxybenzaldehyde-fluctuated greatly, showing no correlation with the pretreatments. Scarification of beans resulted in nearly 4- and 3.6-fold higher vanillin formations on the 10th day in NAA- and Ethrel-treated beans, respectively, as compared to control with a significant change in texture. When activities of major relevant enzymes were followed, addition of NAA or Ethrel helped to retain higher levels of cellulase throughout the curing period and higher levels of beta-glucosidase on the 20th day that correlated with higher vanillin content during curing and subsequent periods. Peroxidase, being highest throughout, did not correlate with the change in levels of major flavoring compounds. The pretreatment methods of the present study may find importance for realizing higher flavor formation in a shorter period because the major quality parameters were found to be comparable to those of a commercial sample.

  12. Effect of pretreating of host oil on coprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Hajdu, P.E.; Tierney, J.W.; Wender, I. [Univ. of Pittsburgh, PA (United States)

    1995-12-31

    The principal objective of this research was to determine if coprocessing performance (i.e., coal conversion and oil yield) could be significantly improved by pretreating the heavy resid prior to reacting it with coal. For this purpose, two petroleum vacuum resids (1000{degrees}F+), one from the Amoco Co. and another from the Citgo Co., were used as such and after they had been pretreated by catalytic hydrogenation and hydrocracking reactions. The pretreatments were aimed at improving the host oil by; (1) converting any aromatic structures in the petroleum to hydroaromatic compounds capable of donating hydrogen, (2) cracking the heavy oil to lower molecular weight material that might serve as a better solvent, (3) reducing the coking propensity of the heavy oil through the hydrogenation of polynuclear aromatic compounds, and (4) removing metals and heteroatoms that might poison a coprocessing catalyst. Highly dispersed catalysts, including fine particle Fe- and Mo-based, and dicobalt octacarbonyl, Co{sub 2}(CO){sub 8}, were used in this study. The untreated and pretreated resids were extensively characterized in order to determine chemical changes brought about by the pretreatments. The modified heavy oils were then coprocessed with an Illinois No. 6 coal as well as with a Wyodak coal, and compared to coprocessing with untreated resids under the same hydroliquefaction conditions. The amount of oil derived from coal was estimated by measuring the level of phenolic oxygen (derived mainly from coal) present in the oil products. Results are presented and discussed.

  13. Breakdown of cell wall nanostructure in dilute acid pretreated biomass.

    Science.gov (United States)

    Pingali, Sai Venkatesh; Urban, Volker S; Heller, William T; McGaughey, Joseph; O'Neill, Hugh; Foston, Marcus; Myles, Dean A; Ragauskas, Arthur; Evans, Barbara R

    2010-09-13

    The generation of bioethanol from lignocellulosic biomass holds great promise for renewable and clean energy production. A better understanding of the complex mechanisms of lignocellulose breakdown during various pretreatment methods is needed to realize this potential in a cost and energy efficient way. Here we use small-angle neutron scattering (SANS) to characterize morphological changes in switchgrass lignocellulose across molecular to submicrometer length scales resulting from the industrially relevant dilute acid pretreatment method. Our results demonstrate that dilute acid pretreatment increases the cross-sectional radius of the crystalline cellulose fibril. This change is accompanied by removal of hemicellulose and the formation of R(g) ∼ 135 A lignin aggregates. The structural signature of smooth cell wall surfaces is observed at length scales larger than 1000 A, and it remains remarkably invariable during pretreatment. This study elucidates the interplay of the different biomolecular components in the breakdown process of switchgrass by dilute acid pretreatment. The results are important for the development of efficient strategies of biomass to biofuel conversion.

  14. Ohmic heating pretreatment of algal slurry for production of biodiesel.

    Science.gov (United States)

    Yodsuwan, Natthawut; Kamonpatana, Pitiya; Chisti, Yusuf; Sirisansaneeyakul, Sarote

    2018-02-10

    Suspensions of the model microalga Chlorella sp. TISTR 8990 were pretreated by ohmic heating to facilitate release of lipids from the cells in subsequent extraction and lipase-mediated transesterification to biodiesel. After ohmic pretreatment, the moist biomass was suspended in a system of water, hexane, methanol and immobilized lipase for extraction of lipids and simultaneous conversion to biodiesel. The ohmic pretreatment was optimized using an experimental design based on Taguchi method to provide treated biomass that maximized the biodiesel yield in subsequent extraction-transesterification operation. The experimental factors were the frequency of electric current (5-10 5  Hz), the processing temperature (50-70 °C), the algal biomass concentration in the slurry (algal fresh weight to water mass ratio of 1-3) and the incubation time (1-3 min). Extraction-transesterification of the pretreated biomass was carried out at 40 °C for 24 h using a reaction systems of a fixed composition (i.e. biomass, hexane, methanol, water and immobilized enzyme). Compared to control (i.e. untreated biomass), the ohmic pretreatment under optimal conditions (5 Hz current frequency, 70 °C, 1:2 mass ratio of biomass to water, incubation time of 2-min) increased the rate of subsequent transesterification by nearly 2-fold. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Reduction of erythema in hairless guinea pigs after cutaneous sulfur mustard vapor exposure by pretreatment with niacinamide, promethazine and indomethacin

    Energy Technology Data Exchange (ETDEWEB)

    Yourick, J.J.; Dawson, J.S.; Mitcheltree, L.W.

    1995-12-31

    Erythema is the initial symptom that occurs after sulfur mustard (HD) cutaneous exposure. The time course of HD-induced erythema is similar to that observed after UV irradiation, which can be reduced by indomethacin. Sulfur mustard lethality is decreased by using promethazine, which is an antihistamine. Niacinamide can reduce microvesication after HD vapor exposure in hairless guinea pig (HGP) skin. The present study examines the effect of the combined administration of niacinamide, indomethacin and promethazine used alone or in all possible combinations on the degree of erythema and histopathologic skin damage after HD exposure in HGP. Niacinamide (750 mg kg%`, i.p.), promethazine (12.5 mg kg%1, i.m.) or indomethacin (4 mg kg%1, p.o.) used singly or in combination was given as a 30-min pretreatment before an 8-min HD vapor cup skin exposure. Using a combination pretreatment of niacinamide, promethazine and indomethacin, erythema was reduced at 4 (91%) and 6 (55%) h, but not 24 h after HD. The incidence of histopathological skin changes (microvesicles, follicular involvement, epidermal necrosis, intracellular edema and pustular epidermatitis) 24 h after HD was not reduced. This study indicates that HD (induced erythema) may result from several different mechanisms, including inflammation, histamine release and DNA damage. It is suggested that two phases of inflammation may occur: an early phase sensitive to antihistamines and non-steroidal antiinflammatory drugs and a late phase of extensive cell damage that was not sensitive to these drug pretreatments.

  16. Characterization of human lymphoid cell lines GM9947 and GM9948 as intra- and interlaboratory reference standards for DNA typing

    Energy Technology Data Exchange (ETDEWEB)

    Fregeau, C.J.; Elliott, J.C.; Fourney, R.M. [RCMP Central Forensic Laboratory, Ottawa, Ontario (Canada)] [and others

    1995-07-20

    The incorporation of reference DNA is crucial to the validation of any DNA typing protocol. Currently, reference DNA standards are restricted to molecular size DNA ladders and/or tumor cell line DNA. Either of these, however, presents some limitations. We have rigorously characterized two Epstein-Barr virus (EBV)-immortalized human lymphoid cell lines-GM9947 (female) and GM9948 (male)-to determine their suitability as alternative in-line standards for three widely employed allele profiling strategies. Twenty-one highly polymorphic VNTR-based allelic systems (7 RFLPs, 2 AmpFLPs, and 12 STRs) distributed over 12 chromosomes were scrutinized along with 3 gender-based discriminatory systems. The genetic stability of each locus was confirmed over a period of 225 in vitro population doublings. Allele size estimates and degree of informativeness for each of the 21 VNTR systems were compiled. The reproducibility of allele scoring by traditional RFLP analyses, using both cell lines as reference standards, was also verified by an interlaboratory validation study involving 13 analysts from two geographically distinct forensic laboratories. Taken together, our data indicate that GM9947 and GM9948 genomic DNAs could be adopted as reliable reference standards for DNA typing. 82 refs., 3 figs., 8 tabs.

  17. HPV DNA test

    Science.gov (United States)

    ... HPV testing in women; Cervical cancer - HPV DNA test; Cancer of cervix - HPV DNA test ... The HPV DNA test may be done during a Pap smear . You lie on a table and place your feet in stirrups. The ...

  18. Synthesis of DNA

    Science.gov (United States)

    Mariella, Jr., Raymond P.

    2008-11-18

    A method of synthesizing a desired double-stranded DNA of a predetermined length and of a predetermined sequence. Preselected sequence segments that will complete the desired double-stranded DNA are determined. Preselected segment sequences of DNA that will be used to complete the desired double-stranded DNA are provided. The preselected segment sequences of DNA are assembled to produce the desired double-stranded DNA.

  19. Enzymatic hydrolysis of sugarcane bagasse pretreated with acid or alkali

    Directory of Open Access Journals (Sweden)

    Vivian Cristina Pietrobon

    2011-04-01

    Full Text Available The aim of this study was to evaluate the performance of enzymatic hydrolysis of acid or alkali pretreated sugarcane bagasse for the production of fermentable sugars. The first step consisted of selection of commercial enzymes presenting the highest cellulolytic activities. After selection of four enzymes: HPL, CL, P1 and P4, their performances were tested in the bagasse pretreated with acid and alkali. The sugar content of the hydrolysates was analyzed by anion exchange liquid chromatography. Data showed that the joint action of 0.5% acid pretreatment, 121ºC, 30 minutes and enzyme CL provides the best results, 67.25 g of hexose and 148.13g of pentose per kg of dry bagasse.

  20. Urine pretreatment for waste water processing systems. [for space station

    Science.gov (United States)

    Winkler, H. E.; Verostko, C. E.; Dehner, G. F.

    1983-01-01

    Recovery of high quality water from urine is an essential part of life support on a Space Station to avoid costly launch and resupply penalties. Water can be effectively recovered from urine by distillation following pretreatment by a chemical agent to inhibit microorganism contamination and fix volatile ammonia constituents. This paper presents the results of laboratory investigations of several pretreatment chemicals which were tested at several concentration levels in combination with sulfuric acid in urine. The optimum pretreatment formulation was then evaluated with urine in the Hamilton Standard Thermoelectric Integrated Membrane Evaporation Subsystem (TIMES). Over 2600 hours of test time was accumulated. Results of these laboratory and system tests are presented in this paper.

  1. New pentose dimers with bicyclic moieties from pretreated biomass

    DEFF Research Database (Denmark)

    Rasmussen, H.; Sørensen, Henrik Rokkjær; Tanner, David Ackland

    2017-01-01

    degraded prior to cellulose hydrolysis, a relief in cellulase inhibition was observed, but some inhibition remained, suggesting that other components also played a role in inhibition. We propose that these components include dipentoses with bicyclic moieties and feruloylated tripentoses, because LC......-MS/MS analysis revealed the presence of these components in the liquid from hydrothermal pretreated wheat straw after enzymatic treatment. The reaction mechanisms for synthesis of the new dipentoses having hydroxylated oxane bicyclic residues are considered and they are proposed to be formed as reaction products...... from either xylose or glucose reacting with glyceraldehyde during pretreatment. The data show that the main cellulase inhibition from hydrothermally pretreated wheat straw liquors is due to xylooligosaccharides followed by oligophenolic compounds and the newly discovered dipentose with bicyclic...

  2. Pretreatment attrition and childhood social phobia: Parental concerns about medication.

    Science.gov (United States)

    Young, Brennan J; Beidel, Deborah C; Turner, Samuel M; Ammerman, Robert T; McGraw, Kelly; Coaston, Susannah C

    2006-01-01

    Pretreatment attrition, the systematic self-exclusion of potential participants during the recruitment phase of a study, poses a significant threat to the external validity of randomized clinical trials. Very little is known about the factors that contribute to pretreatment attrition, especially among families seeking treatment for a child. The current study assessed pretreatment attrition in a randomized clinical trial of behavior therapy, fluoxetine, and placebo for child and adolescent social phobia. Reluctance toward medication treatment accounted for 44.7% of study refusals and was disproportionately common among ethnic minority families. Parents were particularly worried about the potential for side effects or physical/psychological dependency upon the medication. Results are discussed in terms of the implications for external validity in future psychopharmacological clinical trials.

  3. Hazard Analysis for the Pretreatment Engineering Platform (PEP)

    Energy Technology Data Exchange (ETDEWEB)

    Sullivan, Robin S.; Geeting, John GH; Lawrence, Wesley E.; Young, Jonathan

    2008-07-10

    The Pretreatment Engineering Platform (PEP) is designed to perform a demonstration on an engineering scale to confirm the Hanford Waste Treatment Plant Pretreatment Facility (PTF) leaching and filtration process equipment design and sludge treatment process. The system will use scaled prototypic equipment to demonstrate sludge water wash, caustic leaching, oxidative leaching, and filtration. Unit operations to be tested include pumping, solids washing, chemical reagent addition and blending, heating, cooling, leaching, filtration, and filter cleaning. In addition, the PEP will evaluate potential design changes to the ultrafiltration process system equipment to potentially enhance leaching and filtration performance as well as overall pretreatment throughput. The skid-mounted system will be installed and operated in the Processing Development Laboratory-West at Pacific Northwest National Laboratory (PNNL) in Richland, Washington.

  4. A xylanase-aided enzymatic pretreatment facilitates cellulose nanofibrillation.

    Science.gov (United States)

    Long, Lingfeng; Tian, Dong; Hu, Jinguang; Wang, Fei; Saddler, Jack

    2017-11-01

    Although biological pretreatment of cellulosic fiber based on endoglucanases has shown some promise to facilitate cellulose nanofibrillation, its efficacy is still limited. In this study, a xylanase-aided endoglucanase pretreatment was assessed on the bleached hardwood and softwood Kraft pulps to facilitate the downstream cellulose nanofibrillation. Four commercial xylanase preparations were compared and the changes of major fiber physicochemical characteristics such as cellulose/hemicellulose content, gross fiber properties, fiber morphologies, cellulose accessibility/degree of polymerization (DP)/crystallinity were systematically evaluated before and after enzymatic pretreatment. It showed that the synergistic cooperation between endoglucanase and certain xylanase (Biobrite) could efficiently "open up" the hardwood Kraft pulp with limited carbohydrates degradation (cellulose nanofibrillation during mild sonication process (90Wh) with more uniform disintegrated nanofibril products (50-150nm, as assessed by scanning electron microscopy and UV-vis spectroscopy). Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Pretreatment of food waste with high voltage pulse discharge towards methane production enhancement.

    Science.gov (United States)

    Zou, Lianpei; Ma, Chaonan; Liu, Jianyong; Li, Mingfei; Ye, Min; Qian, Guangren

    2016-12-01

    Anaerobic batch tests were performed to investigate the methane production enhancement and solid transformation rates from food waste (FW) by high voltage pulse discharge (HVPD) pretreatment. The total cumulative methane production with HVPD pretreatment was 134% higher than that of the control. The final volatile solids transformation rates of FW with and without HVPD pretreatment were 54.3% and 32.3%, respectively. Comparison study on HVPD pretreatment with acid, alkali and ultrasonic pretreatments showed that the methane production and COD removal rates of FW pretreated with HVPD were more than 100% higher than the control, but only about 50% higher can be obtained with other pretreatments. HVPD pretreatment could be a promising pretreatment method in the application of energy recovery from FW. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Pretreatment of woody biomass for biofuel production: energy efficiency, technologies, and recalcitrance.

    Science.gov (United States)

    Zhu, J Y; Pan, Xuejun; Zalesny, Ronald S

    2010-07-01

    This mini review discusses several key technical issues associated with cellulosic ethanol production from woody biomass: energy consumption for woody biomass pretreatment, pretreatment energy efficiency, woody biomass pretreatment technologies, and quantification of woody biomass recalcitrance. Both total sugar yield and pretreatment energy efficiency, defined as the total sugar recovery divided by total energy consumption for pretreatment, should be used to evaluate the performance of a pretreatment process. A post-chemical pretreatment wood size-reduction approach was proposed to significantly reduce energy consumption. The review also emphasizes using a low liquid-to-wood ratio (L/W) to reduce thermal energy consumption for any thermochemical/physical pretreatment in addition to reducing pretreatment temperature.

  7. Deep Eutectic Solvents pretreatment of agro-industrial food waste.

    Science.gov (United States)

    Procentese, Alessandra; Raganati, Francesca; Olivieri, Giuseppe; Russo, Maria Elena; Rehmann, Lars; Marzocchella, Antonio

    2018-01-01

    Waste biomass from agro-food industries are a reliable and readily exploitable resource. From the circular economy point of view, direct residues from these industries exploited for production of fuel/chemicals is a winning issue, because it reduces the environmental/cost impact and improves the eco-sustainability of productions. The present paper reports recent results of deep eutectic solvent (DES) pretreatment on a selected group of the agro-industrial food wastes (AFWs) produced in Europe. In particular, apple residues, potato peels, coffee silverskin, and brewer's spent grains were pretreated with two DESs, (choline chloride-glycerol and choline chloride-ethylene glycol) for fermentable sugar production. Pretreated biomass was enzymatic digested by commercial enzymes to produce fermentable sugars. Operating conditions of the DES pretreatment were changed in wide intervals. The solid to solvent ratio ranged between 1:8 and 1:32, and the temperature between 60 and 150 °C. The DES reaction time was set at 3 h. Optimal operating conditions were: 3 h pretreatment with choline chloride-glycerol at 1:16 biomass to solvent ratio and 115 °C. Moreover, to assess the expected European amount of fermentable sugars from the investigated AFWs, a market analysis was carried out. The overall sugar production was about 217 kt yr -1 , whose main fraction was from the hydrolysis of BSGs pretreated with choline chloride-glycerol DES at the optimal conditions. The reported results boost deep investigation on lignocellulosic biomass using DES. This investigated new class of solvents is easy to prepare, biodegradable and cheaper than ionic liquid. Moreover, they reported good results in terms of sugars' release at mild operating conditions (time, temperature and pressure).

  8. Effect of nitrogen oxide pretreatments on enzymatic hydrolysis of cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Borrevik, R.K.; Wilke, C.R.; Brink, D.L.

    1978-09-01

    This work considers the effect of nitrogen oxide pretreatments on the subsequent enzymatic hydrolysis by Trichoderma viride cellulase of the cellulose occurring in wheat straw; Triticum Aestivum-L, em. Thell. In the pretreatment scheme the straw is first reacted with nitric oxide and air, and then extracted in aqueous solution. In this way, overall sugar yields increased from 17% for the case of no pretreatment to 70%. The glucose yield increased from 20 to 60%. The yield of glucose during enzymatic hydrolysis is dependent on the reaction time of the gas phase reaction. For a 24 hour reaction the yield is 60%, but drops to 45% for a reaction time of 2 hours. Xylose, a potentially valuable side product of the pretreatment, is obtained by dilute acid hydrolysis during the extraction stage in yields of 90 to 96%. In acidic media, the kinetics of both the rate of formation and destruction of xylose were found to follow the first-order rate laws reported in the literature. These were determined to be 4.5 (liter/gmole)(hr./sup -1/) and 0.03 hr./sup -1/, respectively. However, the rate of formation is much greater (20.4 (liter/gmole) (hr./sup -1/)) when the extraction liquor is recycled. The most likely explanation for this is that the increased total acidity of the recycled liquor compensates for diffusional limitations. A preliminary design and cost analysis of the pretreatment-hydrolysis scheme indicates that glucose can be produced at 10.86 cents per pound, exclusive of straw cost. The corresponding cost per pound of total sugars produced is 5.0 cents. Sensitivity analyses indicate that 42% of the pretreatment cost (excluding hydrolysis) can be attributed to nitric oxide production, and the high yield of sugar obtained is advantageous when considering the cost of straw.

  9. Fungal pretreatment of straw for enhanced biogas yield

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Xinmei; Pilar Castillo, Maria del; Schnuerer, Anna

    2013-07-01

    Among lignocellulosic materials from the agricultural sector, straw is considered to have the biggest potential as a biofuel and therefore also represents a big potential for biogas production. However, the degradation of lignocellulosic materials is somewhat restricted due to the high content of lignin that binds cellulose and hemicellulose and makes them unavailable for microbial degradation. Consequently, low methane yields are achieved. The biodegradability of the lignocellulosic material can be increased by a pretreatment. Optimally the pre-treatment should give an increase in the formation of sugars while avoiding the degradation or loss of carbohydrates and the formation of inhibitory by-products. The treatment should also be cost-effective. Different methods for pre-treatment of lignocellulosic material have been explored, for example thermal, acid, alkaline and oxidative pretreatments. However, they often have a high energy demand. Biological treatment with fungi represents an alternative method for pretreatment of lignocellulosic materials that could be comparably more environmentally friendly, easier to operate and with low energy input. The fungal groups of interest for lignocellulose degradation are the wood decaying fungi, such as the white-, brown-rot and cellulose degraders. The purpose with this work was to increase the biogas potential of straw by using a pretreatment with fungi. Straw was incubated with fungi at aerobic conditions under certain periods of time. The growth and colonization of the straw by the fungi was expected to increase the availability of the lignocellulosic structure of the straw and thus positively affect the biogas potential. In addition also, the spent lignocellulosic material from the cultivation of edible fungi was investigated. We hypothesized that also growth of edible fungi could give a more accessible material and thus give higher biogas potential compared to the substrate before fungal growth.

  10. Bioethanol production: Pretreatment and enzymatic hydrolysis of softwood

    Energy Technology Data Exchange (ETDEWEB)

    Tengborg, Charlotte

    2000-05-01

    The enzymatic hydrolysis process can be used to produce bioethanol from softwood, which are the dominating raw material in the Northern hemisphere. This thesis deals with the development of the process focusing on the pretreatment and the enzymatic hydrolysis stages. The influence of pretreatment conditions on sugar yield, and the effect of inhibitors on the ethanol yield, were investigated for spruce and pine. The maximum yields of hemicellulose sugars and glucose were obtained under different pretreatment conditions. This indicates that two-stage pretreatment may be preferable. The added catalysts, H{sub 2}SO{sub 4} and SO{sub 2}, resulted in similar total sugar yields about 40 g/100 g dry raw material. However, the fermentability of SO{sub 2}-impregnated material was better. This pretreatment resulted in the formation of inhibitors to the subsequent process steps, e.g. sugar and lignin degradation products. The glucose yield in the enzymatic hydrolysis stage was affected by various parameters such as enzyme loading, temperature, pH, residence time, substrate concentration, and agitation. To decrease the amount of fresh water used and thereby waste water produced, the sugar-rich prehydrolysate from the pretreatment step was included in the enzymatic hydrolysis of the solid fraction, resulting in a reduction in the cellulose conversion of up to 36%. Different prehydrolysate detoxification methods, such as treatment with Ca(OH){sub 2}, laccase, and fermentation using yeast, were investigated. The latter was shown to be very efficient. The amount of fresh water used can be further reduced by recycling various process streams. This was simulated experimentally in a bench-scale process. A reduction in fresh water demand of 50% was obtained without any further negative effects on either hydrolysis or fermentation.

  11. Pre-treating Seed to Enhance Germination of Desert Shrubs

    Energy Technology Data Exchange (ETDEWEB)

    W. K. Ostler; D. C. Anderson; D. J. Hansen

    2002-06-01

    Creosotebush [Larrea tridentata (D.C.) Cav.] and white bursage [Ambrosia dumosa (A. Gray) W.W. Payne] seeds were subjected to pre-treatments of rinsing and soaking in water and thiourea to enhance germination in laboratory experiments. The effects of darkness, temperature, seed source, and soil moisture were also evaluated in the laboratory. The best pre-treatment from the laboratory experiments, rinsing with water for 36 hours followed by drying, was field-tested at Fort Irwin, California. Two sites and two seeding dates (early March and mid April) were determined for each site. Five mulch treatments (no mulch, straw, gravel, chemical stabilizer, and plastic) were evaluated in combination with the seed pre-treatments. Field emergence was greatly enhanced with the seed pre-treatment for white bursage during the March (18-42% increase in germination) and April seedings (16-23% increase in germination). Creosotebush showed poor germination during March (2-5%) when soil temperatures averaged 15 C, but germination increased during the April trials (6-43%) when soil temperatures averaged 23 C. The seed pre-treatment during the April trials increased germination from 16-23%. The plastic mulch treatment increased germination dramatically during both the March and April trials. The plastic mulch increased soil temperatures (8-10 C)and maintained high humidity during germination. Both the chemical stabilizer and the gravel mulches improved germination over the control while the straw mulch decreased germination. These results suggest that seed pre-treatments combined with irrigation and mulch are effective techniques to establish these two dominant Mojave Desert species from seed.

  12. Partial Protection of PC12 Cells from Cellular Stress by Low-Dose Sodium Nitroprusside Pre-treatment.

    Science.gov (United States)

    Varga, Judit; Bátor, Judit; Nádasdi, Gergő; Árvai, Zita; Schipp, Renáta; Szeberényi, József

    2016-10-01

    The PC12 rat pheochromocytoma cell line is an in vitro model system widely used for the investigation of intracellular signaling events contributing to neuronal differentiation and cell death. We found earlier that the nitric oxide donor compound sodium nitroprusside (SNP) induced apoptosis of PC12 cells if it was applied in high concentration (400 µM). Yoshioka et al. (J Pharmacol Sci 101:126-134, 2006) reported that cell death evoked by cytotoxic concentrations of SNP could be prevented by a 100 µM SNP pre-treatment in a murine macrophage cell line. The apoptosis caused by toxic-dose SNP treatment (400 µM) could be partially overcome in PC12 cells as well by the low-dose SNP pre-treatment. The partial inhibition of apoptosis was accompanied by reduced phosphorylation of certain proteins (such as stress-activated protein kinases, the p53, and the eIF2α proteins), decreased caspase activation, and less intense internucleosomal DNA fragmentation. The 100 µM SNP pre-treatment reduced the pro-apoptotic potential of certain other stress stimuli (serum withdrawal, cisplatin and tunicamycin treatments) as well, although the underlying biochemical changes were not entirely uniform. On the contrary, the 100 µM SNP pre-treatment was unable to prevent cell death caused by the protein synthesis inhibitor anisomycin. Further clarification of the above-mentioned processes may be important in understanding the mechanisms by which mild nitrosative stress protects cells against certain forms of cellular stress conditions.

  13. Pre-treated bioactive composite in rat soft tissue

    Energy Technology Data Exchange (ETDEWEB)

    Tirri, T.; Jaakkola, T.; Naerhi, T.; Yli-Urpo, A. [Turku Univ. (Finland). Biomaterials Research and Inst. of Dentistry; Rich, J.; Seppaelae, J. [Helsinki Univ. of Technology, Espoo (Finland). Dept. of Chemical Technology

    2001-07-01

    Effect of in vitro formed calcium phosphate surface on a bioactive composite was studied in rat subcutaneous tissue. Pre-treatment in simulated body fluid (SBF) for 14 days resulted in the formation of calcium phosphate deposites on the composite surface whereas no formation was observed on the copolymer without bioactive glass. Pre-treatment had no effect on short term soft tissue reactions around the copolymer without bioactive glass granules whereas the calcium phosphate surface formed on the composite resulted in delayed healing of the surgical wound. This may be due to mechanical stress caused by rough calcium phosphate surface. (orig.)

  14. Cleanser, Detergent, Personal Care Product, and Pretreatment Evaluation

    Science.gov (United States)

    Adam, Niklas; Carrier, Chris; Vega, Leticia; Casteel, Michael; Verostko, chuck; Pickering, Karen

    2011-01-01

    The purpose of the Cleanser, Detergent, Personal Care Product, and Pretreatment Evaluation & Selection task is to identify the optimal combination of personal hygiene products, crew activities, and pretreatment strategies to provide the crew with sustainable life support practices and a comfortable habitat. Minimal energy, mass, and crew time inputs are desired to recycle wastewater during long duration missions. This document will provide a brief background on the work this past year supporting the ELS Distillation Comparison Test, issues regarding use of the hygiene products originally chosen for the test, methods and results used to select alternative products, and lessons learned from testing.

  15. Plasma-Assisted Pretreatment of Wheat Straw for Ethanol Production

    DEFF Research Database (Denmark)

    Schultz-Jensen, Nadja; Kádár, Zsófia; Thomsen, Anne Belinda

    2011-01-01

    , while lignin was degraded up to 95% by O3. The loss of biomass after washing could be explained by the amount of lignin degraded. The washing water of pretreated samples (0–7 h) was analyzed for potential fermentation inhibitors. Approximately 30 lignin degradation products and a number of simple...... carboxylic acids and phenolic compounds were found, e.g., vanillic acid, acetic acid, and formic acid. Some components had the highest concentration at the beginning of the ozonisation process (0.5, 1 h), e.g., 4-hydroxybenzladehyde, while the concentration of others increased during the entire pretreatment...

  16. GSDO Program Hexavalent Chrome Alternatives: Final Pretreatments Test Report

    Science.gov (United States)

    Kessel, Kurt

    2013-01-01

    Hexavalent chrome free pretreatments should be considered for use on Ground Support Equipment (OSE) and Electrical Ground Support Equipment (EOSE). Both of the hexavalent chrome free pretreatments (Metalast TCP HF and SurTec 650C) evaluated by this project met, and in some instances exceeded, the requirements ofMIL-DTL-5541 "Chemical Conversion Coatings on Aluminum and Aluminum Alloys". For DC resistance measurements, both Metalast TCP HF and SurTec (!50C met initial requirements following assembly and in many cases continued to maintain passing readings for the duration of testing.

  17. Epstein-Barr virus associated central nervous system leiomyosarcoma occurring after renal transplantation: case report and review of the literature; Leiomyosarcome primitif du systeme nerveux central associe au virus d'Epstein-Barr (EBV) et survenu apres transplantation renale: a propos d'un cas et revue de la litterature

    Energy Technology Data Exchange (ETDEWEB)

    Tahri, A.; Noel, G.; Feuvret, L.; Jauffret, E.; Brun, B.; Mazeron, J.J.; Baillet, F. [Centre des Tumeurs, Groupe Hospitalier Universitaire Pitie-Salpetriere, 75 - Paris (France); Feuvret, L. [Centre de Protontherapie d' Orsay, 91 (France); Figuerella-Branger, D. [Hopital de la Timone, Service d' Anatomopathologie, 13 - Marseille (France); Goncalves, A. [Institut Paoli-Calmettes, Service d' Oncologie Medicale, 13 - Marseille (France)

    2003-10-01

    Central nervous system leiomyosarcomas are extremely rare, however, they became more frequent among immuno-deficient patients, either in a patients infected with human immunodeficiency virus (HIV), or after organ transplantation. The data of the literature indicate that the infection by Epstein-Barr virus (EBV) plays a causal role in the development of these tumours but its precise role in the onco-genesis remains unresolved. We report a new case of EBV associated leiomyosarcoma of the left cavernous sinus occurring after renal transplantation. The epidemiological, clinical, pathological and therapeutic characteristics of these tumours are discussed. (authors)

  18. Fungal cellulase/xylanase production and corresponding hydrolysis using pretreated corn stover as substrates.

    Science.gov (United States)

    Zhang, Liang; Wang, Xiaoqing; Ruan, Zhenhua; Liu, Ying; Niu, Xiaorui; Yue, Zhengbo; Li, Zhimin; Liao, Wei; Liu, Yan

    2014-01-01

    Three pretreated corn stover (ammonia fiber expansion, dilute acid, and dilute alkali) were used as carbon source to culture Trichoderma reesei Rut C-30 for cellulase and xylanase production. The results indicated that the cultures on ammonia fiber expansion and alkali pretreated corn stover had better enzyme production than the acid pretreated ones. The consequent enzymatic hydrolysis was performed applying fungal enzymes on pretreated corn stover samples. Tukey's statistical comparisons exhibited that there were significant differences on enzymatic hydrolysis among different combination of fungal enzymes and pretreated corn stover. The higher sugar yields were achieved by the enzymatic hydrolysis of dilute alkali pretreated corn stover.

  19. Methods of pretreating comminuted cellulosic material with carbonate-containing solutions

    Energy Technology Data Exchange (ETDEWEB)

    Francis, Raymond

    2012-11-06

    Methods of pretreating comminuted cellulosic material with an acidic solution and then a carbonate-containing solution to produce a pretreated cellulosic material are provided. The pretreated material may then be further treated in a pulping process, for example, a soda-anthraquinone pulping process, to produce a cellulose pulp. The pretreatment solutions may be extracted from the pretreated cellulose material and selectively re-used, for example, with acid or alkali addition, for the pretreatment solutions. The resulting cellulose pulp is characterized by having reduced lignin content and increased yield compared to prior art treatment processes.

  20. Síndrome linfoproliferativo ligado al cromosoma X, infección por el virus EBV y defectos en la regulación de la citotoxicidad linfocitaria X-linked lymphoproliferative syndrome, EBV infection and impaired regulation of cell-mediated cytotoxicity

    Directory of Open Access Journals (Sweden)

    A. Malbrán

    2003-01-01

    Full Text Available La deficiencia del gen SH2D1A que codifica para la proteína reguladora SAP trae aparejada la activación incontrolada de la vía de activación linfocitaria señalizada por SLAM (molécula señaladora de la activación linfocitaria. Es una inmunodeficiencia ligada al cromosoma X (XLP que se pone en evidencia cuando los pacientes portadores de mutaciones en el gen se enfrentan con el virus de Epstein Barr, desarrollando una mononucleosis infecciosa fulminante. Algunos pacientes desarrollan un síndrome linfoproliferativo fatal; los que sobreviven pueden presentar hipogammaglobulinemia severa y mayor frecuencia de neoplasia hematológica que la población normal. En esta revisión se discuten los mecanismos inmuno-regulatorios involucrados en el desarrollo de la patología mencionada, así como la participación de diferentes células efectoras de la respuesta inmune (linfocitos CD8 citotóxicos, células NK.Mutations in SH2D1A, a gene that codifies for the regulatory protein SAP, result in uncontrolled activation of the SLAM (signaling lymphocyte-activation molecule pathway. This X-linked immunodeficiency becomes evident when the patients are infected with Epstein Barr virus (EBV and develop a fulminant form of infectious mononucleosis leading to a lymphoproliferative syndrome that is often fatal (X-linked lymphoproliferative syndrome, XLP. In those who survive, hypogammaglobulinemia and oncohematologic diseases are frequently observed. In this revision, the immuno-regulatory mechanisms involved in XLP immunopathology and the role of different effector cells (CD8 T lymphocytes, NK cells are discussed.

  1. Effect of moisture on pretreatment efficiency for anaerobic digestion of lignocellulosic substrates.

    Science.gov (United States)

    Peces, M; Astals, S; Mata-Alvarez, J

    2015-12-01

    The present study evaluates the effect of moisture in low-temperature and ultrasound pretreatment on lignocellulosic substrates anaerobic biodegradability, where brewer's spent grain was used as model substrate. Besides moisture content, low-temperature pretreatment was also evaluated in terms of temperature (60-80°C) and exposure time (12-72 h). Likewise, ultrasonication was also evaluated in terms of specific energy (1000-50,000 kJ kg TS(-1)). In addition, the effect of substrate particle size reduction by milling pretreatment was also considered. The results clearly demonstrated that substrate moisture (total solid concentration) is a significant parameter for pretreatment performance, although it has been rarely considered in pretreatment optimisation. Specifically, moisture optimisation increased the methane yield of brewer's spent grain by 6% for low-temperature pretreatment (60°C), and by 14% for ultrasound pretreatment (1000 kJ kg TS(-1)) towards the control (without pretreatment). In both pretreatments, the experimental optimum total solid concentration was 100 gTS kg(-1). Thus, lowering substrate moisture, a strategy suggested attaining energetic pretreatment feasibility, needs to be analysed as another pretreatment variable since it might have limited correlation. Finally, a preliminary energetic balance of the pretreatments under study showed that the extra methane production could not cover the energetic pretreatment expenses. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Tank Focus Area Pretreatment Program. FY 1995 Program Management Plan

    International Nuclear Information System (INIS)

    Morrison, M.I.; McGinnis, C.P.; Wilkenson, W.T.; Hunt, R.D.

    1995-02-01

    This program management plan (PMP) describes the FY 1995 project plans for the Pretreatment Program of the Tank Focus Area. The Tank Focus Area is one of five areas of environmental concerns originally identified by the Deputy Assistant Secretary for Technology Development (EM-50). Projects in the Tank Focus Area relate to the remediation of liquid waste stored in underground storage tanks at various US Department of Energy sites. The Pretreatment Program is an organizational unit performing work within the Tank Focus Area. The function of the Pretreatment Program is to develop, test, evaluate, and demonstrate new technologies, with emphasis on separations. The 11 Pretreatment Program projects for FY 1995 are (1) Cesium Extraction Testing, (2) Comprehensive Supernate Treatment, (3) Hot Cell Studies, (4) Cesium Removal Demonstration, (5) Out-of-Tank Evaporator Demonstration, (6) Crossflow Filtration, (7) Technical Interchange with CEA, (8) TRUEX Applications, (9) NAC/NAG Process Studies (conducted at Oak Ridge National Laboratory), (10) NAC/NAG Process and Waste Form Studies (conducted at Florida International University), and (11) Program Management. Section 2 of this PMP contains a separate subsection for each FY 1995 project. A brief description of the project, a schedule of major milestones, and a breakdown of costs are provided for each project. The PMP also contains sections that describe the project controls that are in place. Quality assurance, document control, the project management system, and the management organization are described in these sections

  3. 40 CFR 418.76 - Pretreatment standard for new sources.

    Science.gov (United States)

    2010-07-01

    ... properties controlled by this section which may be discharged to a publicly owned treatment works by a new source subject to the provisions of this subpart: Pollutant or pollutant property Pretreatment standard BOD5 No limitations. TSS Do. pH Do. Ammonia (as N) 30 mg/l. Nitrate (as N) Do. Total phosphorus (as P...

  4. Bromine pretreated chitosan for adsorption of lead (II) from water

    Indian Academy of Sciences (India)

    Lead; 30% bromine pretreated chitosan; water; adsorption; langmuir; Freundlich. 1. Introduction. Many toxic heavy metals like lead, copper, zinc, mercury, chromium, arsenic and cadmium etc are either contami- nated as natural constituents in earth crust (sediments deposit as metal salts) or enter owing to urbanization and ...

  5. Synergistic Effect of Trehalose and Saccharose Pretreatment on ...

    African Journals Online (AJOL)

    2). Osmotic fragility is an important factor in the maintenance of RBC integrity and normal functions [23]. Trehalose and saccharose pretreatment synergistically decreased lyophilization-rehydration-induced damage on. RBC osmotic fragility by reducing the osmotic fragility (Fig. 3). PS is only distributed in the internal side of ...

  6. Effects of glycyrrhizin pre-treatment on transient ischemic brain ...

    African Journals Online (AJOL)

    Effects of glycyrrhizin pre-treatment on transient ischemic brain injury in mice. ... on transient ischemic brain injury in mice. Chiyeon Lim, Sehyun Lim, Young-Jun Lee, Bokcheul Kong, Byoungho Lee, Chang-Hyun Kim, Buyeo Kim, Suin Cho ... induced brain damage. Keywords: Glycyrrhizin, licorice, stroke, apoptosis ...

  7. Bromine pretreated chitosan for adsorption of lead (II) from water

    Indian Academy of Sciences (India)

    Pollution by heavy metals like lead (II) is responsible for health hazards and environmental degradation. Adsorption is a prevalent method applied for removal of heavy metal pollutants from water. This study explored adsorption performances of 30% bromine pretreated chitosan for lead (II) abatement from water. Bromine ...

  8. Effects of Chloramphenicol Pretreatment on Xylazine/ketamine ...

    African Journals Online (AJOL)

    Keyword: Chloramphenicol, xylazine, ketamine, anaesthesia, cats. The effect of pretreatment with a single intramuscular (im) dose of chloramphenicol (10mg/kg) on the anaethesia induced with im injection of ketamine (25mg/kg) was investigated in five cats premedicated with im xylazine (1.0mg/kg) and atropine ...

  9. The Use of Clay-Polymer Nanocomposites in Wastewater Pretreatment

    Science.gov (United States)

    Rytwo, Giora

    2012-01-01

    Some agricultural effluents are unsuitable for discharge into standard sewage-treatment plants: their pretreatment is necessary to avoid clogging of the filtering devices by colloidal matter. The colloidal stability of the effluents is mainly due to mutual repulsive forces that keep charged particles in suspension. Pretreatment processes are based on two separate stages: (a) neutralization of the charges (“coagulation”) and (b) bridging between several small particles to form larger aggregates that sink, leaving clarified effluent (“flocculation”). The consequent destabilization of the colloidal suspension lowers total suspended solids (TSSs), turbidity, and other environmental quality parameters, making the treatments that follow more efficient. Clay-based materials have been widely used for effluent pretreatment and pollutant removal. This study presents the use of nanocomposites, comprised of an anchoring particle and a polymer, as “coagoflocculants” for the efficient and rapid reduction of TSS and turbidity in wastewater with a high organic load. The use of such particles combines the advantages of coagulant and flocculant by neutralizing the charge of the suspended particles while bridging between them and anchoring them to a denser particle (the clay mineral), enhancing their precipitation. Very rapid and efficient pretreatment is achieved in one single treatment step. PMID:22454607

  10. 40 CFR 406.36 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 406.36 Section 406.36 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Normal Wheat Flour Milling Subcategory § 406.36...

  11. 40 CFR 406.14 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for existing sources. 406.14 Section 406.14 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Corn Wet Milling Subcategory § 406.14...

  12. 40 CFR 406.46 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 406.46 Section 406.46 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Bulgur Wheat Flour Milling Subcategory § 406.46...

  13. 40 CFR 406.24 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for existing sources. 406.24 Section 406.24 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS GRAIN MILLS POINT SOURCE CATEGORY Corn Dry Milling Subcategory § 406.24...

  14. Pretreatment on Corn Stover with Low Concentration of Formic Acid

    DEFF Research Database (Denmark)

    Xu, Jian; Thomsen, Mette Hedegaard; Thomsen, Anne Belinda

    2009-01-01

    Bioethanol derived from lignocellulosic biomass has the potential to replace gasoline. Cellulose is naturally recalcitrant to enzymatic attack, and it also surrounded by the matrix of xylan and lignin, which enhances the recalcitrance. Therefore, lignocellulosic materials must be pretreated to make...

  15. Effect of Pretreatments on Seed Viability During Fruit Development ...

    African Journals Online (AJOL)

    Fiifi Baidoo

    foil and incubated for 4 h after which the embryos were then washed and observed for red colouration. The coloured embryos were counted as viable. Percentage viability from five replicates were recorded (Demir & Ellis, 1992). Seed pretreatments and germination tests. Five seeds from each seed lots of the two harvests ...

  16. Effect of Kaempferol Pretreatment on Myocardial Injury in Rats.

    Science.gov (United States)

    Vishwakarma, Anamika; Singh, Thakur Uttam; Rungsung, Soya; Kumar, Tarun; Kandasamy, Arunvikram; Parida, Subhashree; Lingaraju, Madhu Cholenahalli; Kumar, Ajay; Kumar, Asok; Kumar, Dinesh

    2018-01-20

    The present study was undertaken to evaluate the effect of kaempferol in isoprenaline (ISP)-induced myocardial injury in rats. ISP was administered subcutaneously for two subsequent days to induce myocardial injury. Assessment of myocardial injury was done by estimation of hemodynamic functions, myocardial infarcted area, cardiac injury markers, lipid profile, oxidative stress, pro-inflammatory cytokines and histopathology of heart and liver. Rats pretreated with kaempferol showed reduction in the myocardial infarcted area and heart rate. However, no improvement was observed in change in body weight, mean arterial, systolic and diastolic blood pressure. Kaempferol showed significant decrease in serum LDH, CK-MB, troponin-I and lipid profile. However, highest dose of kaempferol did not reduce the serum triglyceride level. Further, antioxidant enzymes, SOD and catalase, were also higher. However, reduced glutathione, serum SGOT and creatinine did not show any improvement. Kaempferol showed reduction in MDA level. Kaempferol at highest dose showed reduction in pro-MMP-2 expression and MMP-9 level. mRNA expression level of TNF-α was not different in kaempferol-pretreated myocardial injured rats with ISP-alone group. Pretreatment with kaempferol at highest dose showed mild mononuclear infiltration and degenerative changes in heart tissue section of myocardial injured rats. Rats pretreated with kaempferol at higher concentration showed normal cordlike arrangement of hepatocytes with moderate swelling of hepatocytes (vacuolar degeneration) around the central vein. Study suggests that kaempferol attenuated lipid profile, infarcted area and oxidative stress in ISP-induced myocardial injury in rats.

  17. Effect of pretreatments on seed viability during fruit development of ...

    African Journals Online (AJOL)

    Studies to identify the stage at which developing fruits of Irvingia gabonensis (var. excelsa and var. gabonensis), picked from standing trees and/or forest floors, attain maximum viability and germinability were conducted in two harvesting seasons in 2000 and 2001. Some pretreatment methods were used as a means of ...

  18. Separations/pretreatment considerations for Hanford privatization phase 2

    Energy Technology Data Exchange (ETDEWEB)

    Hunt, R.D.; McGinnis, C.P.; Welch, T.D.

    1998-05-01

    The Tank Focus Area is funded to develop, demonstrate, and deploy technologies that will assist in the treatment and closure of its nuclear waste tanks. Pretreatment technologies developed to support the privatization effort by the Department of Energy are reviewed. Advancements in evaporation, solid-liquid separation, sludge treatment, solids controls, sodium management, and radionuclide removal are considered.

  19. Influence of pretreatment of agriculture residues on phytase ...

    African Journals Online (AJOL)

    phytase production. Pretreatment of agriculture residues with water to remove excess inorganic phosphate has significantly enhanced the phytase activity in case of de-oiled rice bran, wheat bran, peanut cake (low and high oil) and coconut cake. Maximum increase of 20.3 times in phytase activity was observed in case of ...

  20. Pretreatment of lignocellulose with biological acid recycling (the Biosulfurol process)

    NARCIS (Netherlands)

    Groenestijn, van J.; Hazewinkel, O.; Bakker, R.R.C.

    2006-01-01

    A biomass pretreatment process is being developed based on contacting lignocellulosic biomass with 70% sulfuric acid and subsequent hydrolysis by adding water. In this process, the hydrolysate can be fermented yielding ethanol, while the sulfuric acid is partly recovered by anion-selective membranes

  1. Effect of lime pre-treatment mellowing duration on some ...

    African Journals Online (AJOL)

    The effect of lime pre-treatment duration on some geotechnical properties of shale treated with cement for use as flexible pavement material was studied. Atterberg's limits, compaction, California bearing ratio (CBR) and unconfined compressive strength (UCS) tests were conducted on the natural shale and shale pre-treated ...

  2. Alkaline pretreatment of Mexican pine residues for bioethanol ...

    African Journals Online (AJOL)

    Alkaline pretreatment of Mexican pine residues for bioethanol production. Claudia Elena Soto Alvarez, Javier López Miranda, Gustavo Pérez Verdín, Mario A. Rodríguez Pérez, Isaías Chairez Hernández ...

  3. Fuel ethanol production from alkaline peroxide pretreated corn stover

    Science.gov (United States)

    Corn stover (CS) has the potential to serve as an abundant low-cost feedstock for production of fuel ethanol. Due to heterogeneous complexity and recalcitrance of lignocellulosic feedstocks, pretreatment is required to break the lignin seal and/or disrupt the structure of crystalline cellulose to in...

  4. Efficient Fuel Pretreatment: Simultaneous Torrefaction and Grinding of Biomass

    DEFF Research Database (Denmark)

    Saleh, Suriyati Binti; Hansen, Brian Brun; Jensen, Peter Arendt

    2013-01-01

    Combining torrefaction and grinding of biomass in one reactor may be an attractive fuel pretreatment process. A combined laboratory torrefaction and ball mill reactor has been constructed for studies of the influence of temperature and residence time on the product yields and particle size...

  5. Influence of pre-treatments on the desorption isotherm ...

    African Journals Online (AJOL)

    Influence of pre-treatments on the desorption isotherm characteristics of plaintain. P-N T Johnson. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT · http://dx.doi.org/10.4314/gjs.v39i1.15851 · AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for ...

  6. Sugar cane bagasse pretreatment: An attempt to enhance the ...

    African Journals Online (AJOL)

    +1.5%NaOH. The pretreatment of bagasse with 2.0% H2O2 along with 1.5% NaOH enhanced the biosynthesis of cellulases by H. insolens. Production rate was also optimized with different parameters like thickness of fermentation medium, ...

  7. The Use of Clay-Polymer Nanocomposites in Wastewater Pretreatment

    Directory of Open Access Journals (Sweden)

    Giora Rytwo

    2012-01-01

    Full Text Available Some agricultural effluents are unsuitable for discharge into standard sewage-treatment plants: their pretreatment is necessary to avoid clogging of the filtering devices by colloidal matter. The colloidal stability of the effluents is mainly due to mutual repulsive forces that keep charged particles in suspension. Pretreatment processes are based on two separate stages: (a neutralization of the charges (“coagulation” and (b bridging between several small particles to form larger aggregates that sink, leaving clarified effluent (“flocculation”. The consequent destabilization of the colloidal suspension lowers total suspended solids (TSSs, turbidity, and other environmental quality parameters, making the treatments that follow more efficient. Clay-based materials have been widely used for effluent pretreatment and pollutant removal. This study presents the use of nanocomposites, comprised of an anchoring particle and a polymer, as “coagoflocculants” for the efficient and rapid reduction of TSS and turbidity in wastewater with a high organic load. The use of such particles combines the advantages of coagulant and flocculant by neutralizing the charge of the suspended particles while bridging between them and anchoring them to a denser particle (the clay mineral, enhancing their precipitation. Very rapid and efficient pretreatment is achieved in one single treatment step.

  8. 40 CFR 405.96 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for new sources. 405.96 Section 405.96 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS DAIRY PRODUCTS PROCESSING POINT SOURCE CATEGORY Condensed Milk Subcategory § 405.96...

  9. 40 CFR 405.94 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Pretreatment standards for existing sources. 405.94 Section 405.94 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS DAIRY PRODUCTS PROCESSING POINT SOURCE CATEGORY Condensed Milk...

  10. Microalgal biomass pretreatment for bioethanol production: a review

    Directory of Open Access Journals (Sweden)

    Jesús Velazquez-Lucio

    2018-03-01

    Full Text Available Biofuels derived from microalgae biomass have received a great deal of attention owing to their high potentials as sustainable alternatives to fossil fuels. Microalgae have a high capacity of CO2 fixation and depending on their growth conditions, they can accumulate different quantities of lipids, proteins, and carbohydrates. Microalgal biomass can, therefore, represent a rich source of fermentable sugars for third generation bioethanol production. The utilization of microalgal carbohydrates for bioethanol production follows three main stages: i pretreatment, ii saccharification, and iii fermentation. One of the most important stages is the pretreatment, which is carried out to increase the accessibility to intracellular sugars, and thus plays an important role in improving the overall efficiency of the bioethanol production process. Diverse types of pretreatments are currently used including chemical, thermal, mechanical, biological, and their combinations, which can promote cell disruption, facilitate extraction, and result in the modification the structure of carbohydrates as well as the production of fermentable sugars. In this review, the different pretreatments used on microalgae biomass for bioethanol production are presented and discussed. Moreover, the methods used for starch and total carbohydrates quantification in microalgae biomass are also briefly presented and compared.

  11. 40 CFR 464.16 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... alternate monitoring) 0.363 0.121 (c) Die Casting Operations. PSNS Pollutant or pollutant property Maximum... GUIDELINES AND STANDARDS METAL MOLDING AND CASTING POINT SOURCE CATEGORY Aluminum Casting Subcategory § 464... CFR part 403 and achieve the following pretreatment standards for new sources. (a) Casting Cleaning...

  12. 40 CFR 464.46 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... 0.093 0.0304 Oil and grease (for alternate monitoring) 1.34 0.446 (b) Die Casting Operations. PSNS... GUIDELINES AND STANDARDS METAL MOLDING AND CASTING POINT SOURCE CATEGORY Zinc Casting Subcategory § 464.46... and achieve the following pretreatment standards for new sources. (a) Casting Quench Operations. PSNS...

  13. 40 CFR 408.64 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... sources. Any existing source subject to this subpart that introduces process wastewater pollutants into a... provisions of this subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. Oil and grease Do. TSS Do. [40 FR 6438, Feb. 11, 1975, as amended at 60 FR 33941, June 29, 1995] ...

  14. 40 CFR 408.274 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55794, Dec. 1, 1975, as amended at 60 FR 33947, June 29, 1995] ...

  15. 40 CFR 408.266 - Pretreatment standards for new sources.

    Science.gov (United States)

    2010-07-01

    ... new source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart: Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55793, Dec. 1, 1975, as amended at 60 FR 33947, June 29, 1995] ...

  16. 40 CFR 408.234 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55790, Dec. 1, 1975, as amended at 60 FR 33946, June 29, 1995] ...

  17. 40 CFR 408.164 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... existing source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55781, Dec. 1, 1975, as amended at 60 FR 33944, June 29, 1995] ...

  18. 40 CFR 408.314 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard BOD5 No limitation. TSS Do. pH Do. Oil and grease Do. [40 FR 55798, Dec. 1, 1975, as amended at 60 FR 33948, June 29, 1995] ...

  19. 40 CFR 417.34 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... existing source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do. [40 FR 6442, Feb. 11, 1975, as amended at 60 FR 33952, June 29, 1995] ...

  20. 40 CFR 417.194 - Pretreatment standards for existing sources.

    Science.gov (United States)

    2010-07-01

    ... source subject to this subpart that introduces process wastewater pollutants into a publicly owned... subpart. Pollutant or pollutant property Pretreatment standard pH No limitation. BOD5 Do. TSS Do. Oil and grease Do. COD Do. Surfactants Do. [40 FR 6443, Feb. 11, 1975, as amended at 60 FR 33956, June 29, 1995] ...