18. Process area room. Incinerator to the left. Filter boxes ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

18. Process area room. Incinerator to the left. Filter boxes on the right. Looking north towards change room. - Plutonium Finishing Plant, Waste Incinerator Facility, 200 West Area, Richland, Benton County, WA

Use of the microscreen phage-induction assay to assess the genotoxicity of 14 hazardous industrial wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Houk, V.S.; DeMarini, D.M.

1988-01-01

The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s(lambda), was used to test 14 crude (unfractionated) hazardous industrial waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 pg per ml. Comparisons between the ability of these waste samples to induce prophage and their mutagenicity in the Salmonella reverse mutation assay indicate that the phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, themore » Microscreen assay detected as genotoxic five additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed, as are some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

Assessment of the microscreen phage-induction assay for screening hazardous wastes (1989)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Houk, V.S.; DeMarini, D.M.

1989-01-01

The Microscreen phage-induction assay, which quantitatively measures the induction of prophage Lambda in Escherichia coli WP2s(Lambda), was used to test 14 crude (unfractionated) hazardous industrial-waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons of the mutagenic activity of these waste samples in Salmonella and their ability to induce prophage Lambda indicate that the phage-induction assay was a more-sensitive indicator of genetic damage for this group of wastes. All but one of the wastes that weremore » mutagenic to Salmonella were detected by the phage-induction assay, and 5 wastes not mutagenic to Salmonella were genetically active in the phage assay. The enhanced ability of the phage-induction assay to detect genotoxic activity may be related to the constituents comprising these waste samples. Partial chemical characterizations of the wastes showed high concentrations of carcinogenic metals, solvents, and chlorinated compounds, most of which are detected poorly by the Salmonella assay.« less

Assaying Auxin Receptor Activity Using SPR Assays with F-Box Proteins and Aux/IAA Degrons.

PubMed

Quareshy, Mussa; Uzunova, Veselina; Prusinska, Justyna M; Napier, Richard M

2017-01-01

The identification of TIR1 as an auxin receptor combined with advanced biophysical instrumentation has led to the development of real-time activity assays for auxins. Traditionally, molecules have been assessed for auxinic activity using bioassays, and agrochemical compound discovery continues to be based on "spray and pray" technologies. Here, we describe the methodology behind an SPR-based assay that uses TIR1 and related F-box proteins with surface plasmon resonance spectrometry for rapid compound screening. In addition, methods for collecting kinetic binding data and data processing are given so that they may support programs for rational design of novel auxin ligands.

40 CFR 62.14431 - What must my waste management plan include?

Code of Federal Regulations, 2013 CFR

2013-07-01

.... In developing your waste management plan, you must consider the American Hospital Association (AHA... publication (AHA Catalog Number 057007) is available for purchase from AHA Services, Inc., Post Office Box...

40 CFR 62.14431 - What must my waste management plan include?

Code of Federal Regulations, 2014 CFR

2014-07-01

.... In developing your waste management plan, you must consider the American Hospital Association (AHA... publication (AHA Catalog Number 057007) is available for purchase from AHA Services, Inc., Post Office Box...

8. Front (east) side of incinerator and glove boxes. Ash ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

8. Front (east) side of incinerator and glove boxes. Ash canning hood to the left, combustion chamber in the middle, incinerator hood to the right. Looking west. - Plutonium Finishing Plant, Waste Incinerator Facility, 200 West Area, Richland, Benton County, WA

miR-128 modulates chemosensitivity and invasion of prostate cancer cells through targeting ZEB1.

PubMed

Sun, Xianglun; Li, Youkong; Yu, Jie; Pei, Hong; Luo, Pengcheng; Zhang, Jie

2015-05-01

Recent reports strongly suggest the profound role of miRNAs in cancer therapeutic response and progression, including invasion and metastasis. The sensitivity to therapy and invasion is the major obstacle for successful treatment in prostate cancer. We aimed to investigate the regulative effect of miR-128/zinc-finger E-box-binding homeobox 1 axis on prostate cancer cell chemosensitivity and invasion. The miR-128 expression pattern of prostate cancer cell lines and tissues was detected by real-time reverse transcriptase-polymerase chain reaction, while the mRNA and protein expression levels of zinc-finger E-box-binding homeobox 1 were measured by real-time reverse transcriptase-polymerase chain reaction and western blot assay, respectively. Dual-luciferase reporter gene assay was used to find the direct target of miR-128. Furthermore, prostate cancer cells were treated with miR-128 mimic or zinc-finger E-box-binding homeobox 1-siRNA, and then the cells' chemosensitivity and invasion were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and transwell assay, respectively. We found miR-128 expression obviously decreased in prostate cancer tissues compared with paired normal tissues. Restored miR-128 expression sensitized prostate cancer cells to cisplatin and inhibited the invasion. Furthermore, there was an inverse expression pattern between miR-128 and zinc-finger E-box-binding homeobox 1 in prostate cancer cells and tissues, and zinc-finger E-box-binding homeobox 1 was identified as a direct target of miR-128 in prostate cancer. Knockdown of zinc-finger E-box-binding homeobox 1 expression efficiently sensitized prostate cancer cells to cisplatin and inhibited the invasion. However, ectopic zinc-finger E-box-binding homeobox 1 expression impaired the effects of miR-128 on chemosensitivity and invasion in prostate cancer cells. miR-128 functions as a potential cancer suppressor in prostate cancer progression and rational therapeutic strategies for prostate cancer would be developed based on miR-128/zinc-finger E-box-binding homeobox 1 axis. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Standard Waste Box Lid Screw Removal Option Testing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anast, Kurt Roy

This report provides results from test work conducted to resolve the removal of screws securing the standard waste box (SWB) lids that hold the remediated nitrate salt (RNS) drums. The test work evaluated equipment and process alternatives for removing the 42 screws that hold the SWB lid in place. The screws were secured with a red Loctite thread locker that makes removal very difficult because the rivets that the screw threads into would slip before the screw could be freed from the rivet, making it impossible to remove the screw and therefore the SWB lid.

Arabidopsis F-box protein containing a Nictaba-related lectin domain interacts with N-acetyllactosamine structures.

PubMed

Stefanowicz, Karolina; Lannoo, Nausicaä; Proost, Paul; Van Damme, Els J M

2012-01-01

The Arabidopsis thaliana genome contains a small group of bipartite F-box proteins, consisting of an N-terminal F-box domain and a C-terminal domain sharing sequence similarity with Nictaba, the jasmonate-induced glycan-binding protein (lectin) from tobacco. Based on the high sequence similarity between the C-terminal domain of these proteins and Nictaba, the hypothesis was put forward that the so-called F-box-Nictaba proteins possess carbohydrate-binding activity and accordingly can be considered functional homologs of the mammalian sugar-binding F-box or Fbs proteins which are involved in proteasomal degradation of glycoproteins. To obtain experimental evidence for the carbohydrate-binding activity and specificity of the A. thaliana F-box-Nictaba proteins, both the complete F-box-Nictaba sequence of one selected Arabidopsis F-box protein (in casu At2g02360) as well as the Nictaba-like domain only were expressed in Pichia pastoris and analyzed by affinity chromatography, agglutination assays and glycan micro-array binding assays. These results demonstrated that the C-terminal Nictaba-like domain provides the F-box-protein with a carbohydrate-binding activity that is specifically directed against N- and O-glycans containing N-acetyllactosamine (Galβ1-3GlcNAc and Galβ1-4GlcNAc) and poly-N-acetyllactosamine ([Galβ1-4GlcNAc]n) as well as Lewis A (Galβ1-3(Fucα1-4)GlcNAc), Lewis X (Galβ1-4(Fucα1-3)GlcNAc, Lewis Y (Fucα1-2Galβ1-4(Fucα1-3)GlcNAc) and blood type B (Galα1-3(Fucα1-2)Galβ1-3GlcNAc) motifs. Based on these findings one can reasonably conclude that at least the A. thaliana F-box-Nictaba protein encoded by At2g02360 can act as a carbohydrate-binding protein. The results from the glycan array assays revealed differences in sugar-binding specificity between the F-box protein and Nictaba, indicating that the same carbohydrate-binding motif can accommodate unrelated oligosaccharides.

Assessment of the Microscreen phage-induction assay for screening hazardous wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Houk, V.S.; DeMarini, D.M.

1987-09-01

The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s(lambda), was used to test 14 crude (unfractionated) hazardous industrial waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons between the mutagenicity of these waste samples in Salmonella and their ability to induce prophage lambda indicate that the Microscreen phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, the Microscreen assaymore » detected as genotoxic 5 additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed along with some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

Use of the Microscreen phage-induction assay to assess the genotoxicity of 14 hazardous industrial wastes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Houk, V.S.; DeMarini, D.M.

1988-01-01

The Microscreen phage-induction assay, which quantitatively measures the induction of prophage lambda in Escherichia coli WP2s lambda, was used to test 14 crude (unfractionated) hazardous industrial-waste samples for genotoxic activity in the presence and absence of metabolic activation. Eleven of the 14 wastes induced prophage, and induction was observed at concentrations as low as 0.4 picograms per ml. Comparisons between the mutagenicity of these waste samples in Salmonella and their ability to induce prophage lambda indicate that the Microscreen phage-induction assay detected genotoxic activity in all but one of the wastes that were mutagenic in Salmonella. Moreover, the Microscreen assaymore » detected as genotoxic 5 additional wastes that were not detected in the Salmonella assay. The applicability of the Microscreen phage-induction assay for screening hazardous wastes for genotoxic activity is discussed along with some of the problems associated with screening highly toxic wastes containing toxic volatile compounds.« less

Solar Disinfection of MODS Mycobacterial Cultures in Resource-Poor Settings

PubMed Central

Nathavitharana, Ruvandhi; Coronel, Jorge; Moore, David A. J.

2007-01-01

Introduction Safe disposal of TB culture material in which the infectious burden of clinical samples has been greatly amplified is an important challenge in resource-limited settings. The bactericidal capacity of solar cookers has been demonstrated previously for conventional bacteria and contaminated clinical waste. We investigated the use of a simple solar cooker for the sterilization of mycobacterial broth cultures from the microscopic observation drug susceptibility assay (MODS). Methods Simulated TB culture materials were prepared by inoculating 24-well MODS plates with 500 µL of a known concentration of Mycobacterium bovis BCG. In a series of experiments, samples were simultaneously placed inside a box-type solar cooker and control box and removed at timepoints between 15 minutes and 6 hours. Quantitative cultures were performed using retrieved samples to determine sterilization effect. Results All cultures from the control box were positive at or within 1–4 logs of inoculation concentration. Simulated culture plates at concentrations from 103colony-forming-units (CFU)/ml to 107 CFU/ml were completely sterilized after only one hour of cooker exposure, at temperatures between 50–102°C. At 109 CFU/ml (far in excess of diagnostic cultures), it was only possible to recover mycobacterial growth in plates removed after 15 minutes. By 30 minutes all plates were effectively sterilized. Discussion Solar disinfection provides a very effective, safe and low-cost alternative to conventional equipment used for disposal of mycobacterial culture material. Effect of climatic conditions and optimal operating procedure remain to be defined. PMID:17971863

Bilayer Effects of Antimalarial Compounds

PubMed Central

Ramsey, Nicole B.; Andersen, Olaf S.

2015-01-01

Because of the perpetual development of resistance to current therapies for malaria, the Medicines for Malaria Venture developed the Malaria Box to facilitate the drug development process. We tested the 80 most potent compounds from the box for bilayer-mediated effects on membrane protein conformational changes (a measure of likely toxicity) in a gramicidin-based stopped flow fluorescence assay. Among the Malaria Box compounds tested, four compounds altered membrane properties (p< 0.05); MMV007384 stood out as a potent bilayer-perturbing compound that is toxic in many cell-based assays, suggesting that testing for membrane perturbation could help identify toxic compounds. In any case, MMV007384 should be approached with caution, if at all. PMID:26551613

Bilayer Effects of Antimalarial Compounds.

PubMed

Ramsey, Nicole B; Andersen, Olaf S

2015-01-01

Because of the perpetual development of resistance to current therapies for malaria, the Medicines for Malaria Venture developed the Malaria Box to facilitate the drug development process. We tested the 80 most potent compounds from the box for bilayer-mediated effects on membrane protein conformational changes (a measure of likely toxicity) in a gramicidin-based stopped flow fluorescence assay. Among the Malaria Box compounds tested, four compounds altered membrane properties (p< 0.05); MMV007384 stood out as a potent bilayer-perturbing compound that is toxic in many cell-based assays, suggesting that testing for membrane perturbation could help identify toxic compounds. In any case, MMV007384 should be approached with caution, if at all.

Treatment of Bottled Liquid Waste During Remediation of the Hanford 618-10 Burial Ground - 13001

DOE Office of Scientific and Technical Information (OSTI.GOV)

Faulk, Darrin E.; Pearson, Chris M.; Vedder, Barry L.

2013-07-01

A problematic waste form encountered during remediation of the Hanford Site 618-10 burial ground consists of bottled aqueous waste potentially contaminated with regulated metals. The liquid waste requires stabilization prior to landfill disposal. Prior remediation activities at other Hanford burial grounds resulted in a standard process for sampling and analyzing liquid waste using manual methods. Due to the highly dispersible characteristics of alpha contamination, and the potential for shock sensitive chemicals, a different method for bottle processing was needed for the 618-10 burial ground. Discussions with the United States Department of Energy (DOE) and United States Environmental Protection Agency (EPA)more » led to development of a modified approach. The modified approach involves treatment of liquid waste in bottles, up to one gallon per bottle, in a tray or box within the excavation of the remediation site. Bottles are placed in the box, covered with soil and fixative, crushed, and mixed with a Portland cement grout. The potential hazards of the liquid waste preclude sampling prior to treatment. Post treatment verification sampling is performed to demonstrate compliance with land disposal restrictions and disposal facility acceptance criteria. (authors)« less

Characterization of an AGAMOUS-like MADS Box Protein, a Probable Constituent of Flowering and Fruit Ripening Regulatory System in Banana

PubMed Central

Roy Choudhury, Swarup; Roy, Sujit; Nag, Anish; Singh, Sanjay Kumar; Sengupta, Dibyendu N.

2012-01-01

The MADS-box family of genes has been shown to play a significant role in the development of reproductive organs, including dry and fleshy fruits. In this study, the molecular properties of an AGAMOUS like MADS box transcription factor in banana cultivar Giant governor (Musa sp, AAA group, subgroup Cavendish) has been elucidated. We have detected a CArG-box sequence binding AGAMOUS MADS-box protein in banana flower and fruit nuclear extracts in DNA-protein interaction assays. The protein fraction in the DNA-protein complex was analyzed by mass spectrometry and using this information we have obtained the full length cDNA of the corresponding protein. The deduced protein sequence showed ∼95% amino acid sequence homology with MA-MADS5, a MADS-box protein described previously from banana. We have characterized the domains of the identified AGAMOUS MADS-box protein involved in DNA binding and homodimer formation in vitro using full-length and truncated versions of affinity purified recombinant proteins. Furthermore, in order to gain insight about how DNA bending is achieved by this MADS-box factor, we performed circular permutation and phasing analysis using the wild type recombinant protein. The AGAMOUS MADS-box protein identified in this study has been found to predominantly accumulate in the climacteric fruit pulp and also in female flower ovary. In vivo and in vitro assays have revealed specific binding of the identified AGAMOUS MADS-box protein to CArG-box sequence in the promoters of major ripening genes in banana fruit. Overall, the expression patterns of this MADS-box protein in banana female flower ovary and during various phases of fruit ripening along with the interaction of the protein to the CArG-box sequence in the promoters of major ripening genes lead to interesting assumption about the possible involvement of this AGAMOUS MADS-box factor in banana fruit ripening and floral reproductive organ development. PMID:22984496

Characterization of an AGAMOUS-like MADS box protein, a probable constituent of flowering and fruit ripening regulatory system in banana.

PubMed

Roy Choudhury, Swarup; Roy, Sujit; Nag, Anish; Singh, Sanjay Kumar; Sengupta, Dibyendu N

2012-01-01

The MADS-box family of genes has been shown to play a significant role in the development of reproductive organs, including dry and fleshy fruits. In this study, the molecular properties of an AGAMOUS like MADS box transcription factor in banana cultivar Giant governor (Musa sp, AAA group, subgroup Cavendish) has been elucidated. We have detected a CArG-box sequence binding AGAMOUS MADS-box protein in banana flower and fruit nuclear extracts in DNA-protein interaction assays. The protein fraction in the DNA-protein complex was analyzed by mass spectrometry and using this information we have obtained the full length cDNA of the corresponding protein. The deduced protein sequence showed ~95% amino acid sequence homology with MA-MADS5, a MADS-box protein described previously from banana. We have characterized the domains of the identified AGAMOUS MADS-box protein involved in DNA binding and homodimer formation in vitro using full-length and truncated versions of affinity purified recombinant proteins. Furthermore, in order to gain insight about how DNA bending is achieved by this MADS-box factor, we performed circular permutation and phasing analysis using the wild type recombinant protein. The AGAMOUS MADS-box protein identified in this study has been found to predominantly accumulate in the climacteric fruit pulp and also in female flower ovary. In vivo and in vitro assays have revealed specific binding of the identified AGAMOUS MADS-box protein to CArG-box sequence in the promoters of major ripening genes in banana fruit. Overall, the expression patterns of this MADS-box protein in banana female flower ovary and during various phases of fruit ripening along with the interaction of the protein to the CArG-box sequence in the promoters of major ripening genes lead to interesting assumption about the possible involvement of this AGAMOUS MADS-box factor in banana fruit ripening and floral reproductive organ development.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hensley, D.C.

Nuclear Fuel Services sent more than 800 drums of nuclear waste to Oak Ridge National Laboratory, with the majority of the waste packaged into five different waste matrix types. A thorough and complete assay of the waste was performed at both NFS and at ORNL. A detailed comparing of the two assay sets provides valuable. insights into problems encountered in typical assay campaigns, particularly as there is, for the most part, excellent agreement between these two campaigns.

NAIplot: An opensource web tool to visualize neuraminidase inhibitor (NAI) phenotypic susceptibility results using kernel density plots.

PubMed

Lytras, Theodore; Kossyvakis, Athanasios; Mentis, Andreas

2016-02-01

The results of neuraminidase inhibitor (NAI) enzyme inhibition assays are commonly expressed as 50% inhibitory concentration (IC50) fold-change values and presented graphically in box plots (box-and-whisker plots). An alternative and more informative type of graph is the kernel density plot, which we propose should be the preferred one for this purpose. In this paper we discuss the limitations of box plots and the advantages of the kernel density plot, and we present NAIplot, an opensource web application that allows convenient creation of density plots specifically for visualizing the results of NAI enzyme inhibition assays, as well as for general purposes. Copyright © 2015 Elsevier B.V. All rights reserved.

Demonstration of geophysical methods for burial ground geophysical characterization study at the DOE Savannah River site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hasbrouck, J.C.; MacLean, H.D.; Geotech, R.

1996-11-01

Rust Geotech, operating contractor at the U.S. Department of Energy Grand Junction Projects Office (DOE-GJPO), conducted a demonstration of the trench boundary and large-object location capabilities of five nonintrusive geophysical methods in the Low-Level Radioactive Waste Disposal Facility (LLRWDF) at the DOE Savannah River Site (SRS). The plan for Resource Conservation and Recovery Act (RCRA) closure of the SRS LLRWDF specifies inplace compaction of {open_quotes}B-25{close_quotes} metal boxes containing low-level radioactive wastes. The boxes are buried in Engineered Low-Level Trenches (ELLTs) at the facility. To properly guide and control the compaction operation, the coordinates of the trench boundaries must be determinedmore » to an accuracy within 5 feet and the outer edges of the metal boxes in the trenches must be determined to within 2 feet.« less

Applications Where Snap is BPM for Radioactive Waste Assay

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, T.J.

2008-07-01

Historically, the Atomic Weapons Establishment (AWE) at Aldermaston in the United Kingdom (UK), has used a variety of assay techniques to measure the radioactive content of a diverse range of waste packages from decommissioning, operational and legacy sources. The regulator, the Environment Agency in the UK, places conditions and limits on AWE through an authorisation within the Radioactive Substances Act (RSA93). The conditions and limits require Best Practical Means (BPM) measurements to be used to demonstrate compliance with the authorisation. Hence, the assay technique employed needs to achieve a balance between risk of exposure, environmental considerations, technological considerations, health andmore » safety considerations and cost effectiveness, without being grossly disproportionate in terms of money, time or trouble. Recently published work has concluded that the Spectral Non-destructive Assay Platform (SNAP) assay system is BPM for Depleted Uranium (DU) waste assay at AWE (1) and low level plutonium in soft drummed waste, HEPA filters and soils (2-4). The purpose of this paper is to highlight other applications where SNAP represents BPM for radioactive waste assay. This has been done by intercomparison studies of SNAP with other assay techniques, such as Segmented Gamma Scanner (SGS) and Passive Neutron Coincidence Counter (PNCC). It has been concluded that, for a large range of waste packages encountered at AWE, SNAP is BPM. (author)« less

76 FR 35471 - Notice of Lodging of Material Modification to Consent Decree Under the Clean Air Act

Federal Register 2010, 2011, 2012, 2013, 2014

2011-06-17

... and re-use certain condensed waste water streams at the facility. The proposed Second Decree... mailed either electronically to [email protected] or in hard copy to P.O. Box 7611, U.S... obtained by mail from the Department of Justice Consent Decree Library, P.O. Box 7611, Washington, DC 20044...

Status of the waste assay for nonradioactive disposal (WAND) project

NASA Astrophysics Data System (ADS)

Arnone, Gaetano L.; Foster, Lynn A.; Foxx, Charles L.; Hagan, Roland C.; Martin, E. R.; Myers, Steven C.; Parker, Jack L.

1999-01-01

The WAND (Waste Assay for Nonradioactive Disposal) system scans thought-to-be-clean, low-density waste (mostly paper and plastics) to verify the absence of radioactive contaminants at very low-levels. Much of the low-density waste generated in radiologically controlled areas, formally considered `suspect' radioactive, is now disposed more cheaply at the Los Alamos County Landfill as opposed to the LANL Radioactive Waste Landfill.

Overexpression of VpEIFP1, a novel F-box/Kelch-repeat protein from wild Chinese Vitis pseudoreticulata, confers higher tolerance to powdery mildew by inducing thioredoxin z proteolysis.

PubMed

Wang, Jie; Yao, Wenkong; Wang, Lei; Ma, Fuli; Tong, Weihuo; Wang, Chen; Bao, Rui; Jiang, Changyue; Yang, Yazhou; Zhang, Jianxia; Xu, Yan; Wang, Xiping; Zhang, Chaohong; Wang, Yuejin

2017-10-01

An F-box protein (VpEIFP1) induced by Erysiphe necator was isolated from Vitis pseudoreticulata, a wild Chinese grapevine species naturally resistant to powdery mildew (PM). It contains an F-box domain and two Kelch-repeat motifs. Expression profiles indicate the VpEIFP1 is strongly induced at both transcriptional and translational levels by PM infection. A subcellular localisation assay showed that VpEIFP1 is predominantly located in the nucleus and cytoplasm. Overexpression of VpEIFP1 accelerated the accumulation of hydrogen peroxide (H 2 O 2 ) and up-regulated the expressions of ICS2, NPR1 and PR1 involved in defence responses, resulting in suppression of PM germination and growth. As an F-box protein, VpEIFP1 interacts with thioredoxin z (VpTrxz) in the yeast-two-hybrid (Y2H) assay and in the bimolecular fluorescence complementation (BiFC) assay. Decreased amounts of VpTrxz protein in transgenic grapevine leaves overexpressing VpEIFP1 were restored by proteasome inhibitor MG132, implying that VpEIFP1 mediated VpTrxz for degradation through the SCF VpEIFP1 (Skp1-Cullin-F-box) E3 ubiquitin ligase complex. The RNA interference line of VpTrxz showed increased H 2 O 2 accumulation following PM inoculation. We propose VpEIFP1 positively modulates the grapevine defence response to PM by inducing the degradation of VpTrxz via the ubiquitin/26S proteasome system. Copyright © 2017 Elsevier B.V. All rights reserved.

The activation of p38 MAPK primarily contributes to UV-induced RhoB expression by recruiting the c-Jun and p300 to the distal CCAAT box of the RhoB promoter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahn, Jiwon; Department of Microbiology, Chungnam National University, Daejeon 305-764; Choi, Jeong-Hae

2011-06-03

Highlights: {yields} Regulation of transcriptional activation of RhoB is still unclear. {yields} We examine the effect of p38 MAPK inhibition, and c-Jun and RhoB depletion on UV-induced RhoB expression and apoptosis. {yields} We identify the regions of RhoB promoter necessary to confer UV responsiveness using pRhoB-luciferase reporter assays. {yields} c-Jun, ATF2 and p300 are dominantly associated with NF-Y on the distal CCAAT box. {yields} The activation of p38 MAPK primarily contribute to UV-induced RhoB expression by recruiting the c-Jun and p300 proteins on distal CCAAT box of RhoB promoter. -- Abstract: The Ras-related small GTP-binding protein RhoB is rapidly inducedmore » in response to genotoxic stresses caused by ionizing radiation. It is known that UV-induced RhoB expression results from the binding of activating transcription factor 2 (ATF2) via NF-Y to the inverted CCAAT box (-23) of the RhoB promoter. Here, we show that the association of c-Jun with the distal CCAAT box (-72) is primarily involved in UV-induced RhoB expression and p38 MAPK regulated RhoB induction through the distal CCAAT box. UV-induced RhoB expression and apoptosis were markedly attenuated by pretreatment with the p38 MAPK inhibitor. siRNA knockdown of RhoB, ATF2 and c-Jun resulted in decreased RhoB expression and eventually restored the growth of UV-irradiated Jurkat cells. In the reporter assay using luciferase under the RhoB promoter, inhibition of RhoB promoter activity by the p38 inhibitor and knockdown of c-Jun using siRNA occurred through the distal CCAAT box. Immunoprecipitation and DNA affinity protein binding assays revealed the association of c-Jun and p300 via NF-YA and the dissociation of histone deacetylase 1 (HDAC1) via c-Jun recruitment to the CCAAT boxes of the RhoB promoter. These results suggest that the activation of p38 MAPK primarily contributes to UV-induced RhoB expression by recruiting the c-Jun and p300 proteins to the distal CCAAT box of the RhoB promoter in Jurkat cells.« less

Speaker box made of composite particle board based on mushroom growing media waste

NASA Astrophysics Data System (ADS)

Tjahjanti, P. H.; Sutarman, Widodo, E.; Kurniawan, A. R.; Winarno, A. T.; Yani, A.

2017-06-01

This research aimed to use mushroom growing media waste (MGMW) that was added by urea, starch and polyvinyl chloride (PVC) glue as a composite particle board to be used as the material of speaker box manufacture. Physical and mechanical testing of particle board including density, moisture content, thickness swelling after immersion in water, strength in water absorption, internal bonding, modulus of elasticity, modulus of rupture and screw holding power, were carried out in accordance with the Stándar Nasional Indonesia (SNI) 03-2105-2006 and Japanese International Standard (JIS) A 5908-2003. The optimum composition of composite particle boards was 60% MGMW + 39% (50% urea +50% starch) + 1% PVC glue. Furthermore, the optimum composition to create speaker box with hardness values of 14.9 Brinnel Hardness Number and results of vibration test obtained amplitude values of the Z-axis, minimum of 0.032007 and maximum of 0.151575. For the acoustic test, results showed good sound absorption coefficients at frequencies of 500 Hz and it has better damping absorption.

48 CFR 52.204-4 - Printed or Copied Double-Sided on Postconsumer Fiber Content Paper.

Code of Federal Regulations, 2013 CFR

2013-10-01

... boxes; old newspapers; old magazines; mixed waste paper; tabulating cards; and used cordage; or (2) All paper, paperboard, and fibrous materials that enter and are collected from municipal solid waste; but... practicable, when not using electronic commerce methods to submit information or data to the Government. (End...

48 CFR 52.204-4 - Printed or Copied Double-Sided on Postconsumer Fiber Content Paper.

Code of Federal Regulations, 2012 CFR

2012-10-01

... boxes; old newspapers; old magazines; mixed waste paper; tabulating cards; and used cordage; or (2) All paper, paperboard, and fibrous materials that enter and are collected from municipal solid waste; but... practicable, when not using electronic commerce methods to submit information or data to the Government. (End...

48 CFR 52.204-4 - Printed or Copied Double-Sided on Postconsumer Fiber Content Paper.

Code of Federal Regulations, 2014 CFR

2014-10-01

... boxes; old newspapers; old magazines; mixed waste paper; tabulating cards; and used cordage; or (2) All paper, paperboard, and fibrous materials that enter and are collected from municipal solid waste; but... practicable, when not using electronic commerce methods to submit information or data to the Government. (End...

Comparison of solid-phase and eluate assays to gauge the ecotoxicological risk of organic wastes on soil organisms.

PubMed

Domene, Xavier; Alcañiz, Josep M; Andrés, Pilar

2008-02-01

Development of methodologies to assess the safety of reusing polluted organic wastes in soil is a priority in Europe. In this study, and coupled with chemical analysis, seven organic wastes were subjected to different aquatic and soil bioassays. Tests were carried out with solid-phase waste and three different waste eluates (water, methanol, and dichloromethane). Solid-phase assays were indicated as the most suitable for waste testing not only in terms of relevance for real situations, but also because toxicity in eluates was generally not representative of the chronic effects in solid-phase. No general correlations were found between toxicity and waste pollutant burden, neither in solid-phase nor in eluate assays, showing the inability of chemical methods to predict the ecotoxicological risks of wastes. On the contrary, several physicochemical parameters reflecting the degree of low organic matter stability in wastes were the main contributors to the acute toxicity seen in collembolans and daphnids.

Cleanup Verification Package for the 118-F-1 Burial Ground

DOE Office of Scientific and Technical Information (OSTI.GOV)

E. J. Farris and H. M. Sulloway

2008-01-10

This cleanup verification package documents completion of remedial action for the 118-F-1 Burial Ground on the Hanford Site. This burial ground is a combination of two locations formerly called Minor Construction Burial Ground No. 2 and Solid Waste Burial Ground No. 2. This waste site received radioactive equipment and other miscellaneous waste from 105-F Reactor operations, including dummy elements and irradiated process tubing; gun barrel tips, steel sleeves, and metal chips removed from the reactor; filter boxes containing reactor graphite chips; and miscellaneous construction solid waste.

High mobility group box (HMGB) proteins of Plasmodium falciparum: DNA binding proteins with pro-inflammatory activity.

PubMed

Kumar, Krishan; Singal, Ankita; Rizvi, M Moshahid A; Chauhan, Virander S

2008-06-01

High mobility group box chromosomal protein 1 (HMGB1), known as an abundant, non-histone architectural chromosomal protein, is highly conserved across different species. Homologues of HMGB1 were identified and cloned from malaria parasite, Plasmodium falciparum. Sequence analyses showed that the P. falciparum HMGB1 (PfHMGB1) exhibits 45, 23 and 18%, while PfHMGB2 shares 42, 21 and 17% homology with Saccharomyces cerevisiae, human and mouse HMG box proteins respectively. Parasite PfHMGB1and PfHMGB2 proteins contain one HMG Box domain similar to B-Box of mammalian HMGB1. Electrophoretic Mobility Shift Assay (EMSA) showed that recombinant PfHMGB1 and PfHMGB2 bind to DNA. Immunofluorescence Assay using specific antibodies revealed that these proteins are expressed abundantly in the ring stage nuclei. Significant levels of PfHMGB1 and PfHMGB2 were also present in the parasite cytosol at trophozoite and schizont stages. Both, PfHMGB1 and PfHMGB2 were found to be potent inducers of pro-inflammatory cytokines such as TNFalpha from mouse peritoneal macrophages as analyzed by both reverse transcription PCR and by ELISA. These results suggest that secreted PfHMGB1 and PfHMGB2 may be responsible for eliciting/ triggering host inflammatory immune responses associated with malaria infection.

40 CFR 60.3062 - What is an air curtain incinerator?

Code of Federal Regulations, 2011 CFR

2011-07-01

... curtain of air across an open, integrated combustion chamber (fire box) or open pit or trench (trench... 40 Protection of Environment 6 2011-07-01 2011-07-01 false What is an air curtain incinerator? 60... Rule-Air Curtain Incinerators That Burn Only Wood Waste, Clean Lumber, and Yard Waste § 60.3062 What is...

75 FR 62120 - Notice of Availability of Draft Basis for Determination Under Section 3116 of the Ronald W...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-07

... waste storage tanks and supporting ancillary structures. Two of those waste tanks, Tanks 17 and 20 were... available for public review and comment. DATES: The comment period will end on January 7, 2011. Comments... structures are two evaporator systems, transfer lines, six diversion boxes, one catch tank, a concentrate...

A batch assay to measure microbial hydrogen sulfide production from sulfur-containing solid wastes.

PubMed

Sun, Mei; Sun, Wenjie; Barlaz, Morton A

2016-05-01

Large volumes of sulfur-containing wastes enter municipal solid waste landfills each year. Under the anaerobic conditions that prevail in landfills, oxidized forms of sulfur, primarily sulfate, are converted to sulfide. Hydrogen sulfide (H2S) is corrosive to landfill gas collection and treatment systems, and its presence in landfill gas often necessitates the installation of expensive removal systems. For landfill operators to understand the cost of managing sulfur-containing wastes, an estimate of the H2S production potential is needed. The objective of this study was to develop and demonstrate a biochemical sulfide potential (BSP) test to measure the amount of H2S produced by different types of sulfur-containing wastes in a relatively fast (30days) and inexpensive (125mL serum bottles) batch assay. This study confirmed the toxic effect of H2S on both sulfate reduction and methane production in batch systems, and demonstrated that removing accumulated H2S by base adsorption was effective for mitigating inhibition. H2S production potentials of coal combustion fly ash, flue gas desulfurization residual, municipal solid waste combustion ash, and construction and demolition waste were determined in BSP assays. After 30days of incubation, most of the sulfate in the wastes was converted to gaseous or aqueous phase sulfide, with BSPs ranging from 0.8 to 58.8mLH2S/g waste, depending on the chemical composition of the samples. Selected samples contained solid phase sulfide which contributed to the measured H2S yield. A 60day incubation in selected samples resulted in 39-86% additional sulfide production. H2S production measured in BSP assays was compared with that measured in simulated landfill reactors and that calculated from chemical analyses. H2S production in BSP assays and in reactors was lower than the stoichiometric values calculated from chemical composition for all wastes tested, demonstrating the importance of assays to estimate the microbial sulfide production potential of sulfur-containing wastes. Copyright © 2016 Elsevier B.V. All rights reserved.

29 CFR 1915.91 - Housekeeping.

Code of Federal Regulations, 2011 CFR

2011-07-01

...) All working areas on or immediately surrounding vessels and dry docks, graving docks, or marine... boxes or fire-extinguishing equipment. (e) All oils, paints thinners, solvents, waste, rags, or other...

Pharmaceutical wastewater being composite mixture of environmental pollutants may be associated with mutagenicity and genotoxicity.

PubMed

Sharif, Ali; Ashraf, Muhammad; Anjum, Aftab Ahmed; Javeed, Aqeel; Altaf, Imran; Akhtar, Muhammad Furqan; Abbas, Mateen; Akhtar, Bushra; Saleem, Ammara

2016-02-01

Pharmaceutical industries are amongst the foremost contributor to industrial waste. Ecological well-being is endangered owing to its facile discharge. In the present study, heavy metals and organic contaminants in waste water were characterized using atomic absorption spectrophotometer and GC-MS, respectively. Mutagenicity and genotoxic potential of pharmaceutical waste water were investigated through bacterial reverse mutation assay and in vitro comet assay, respectively. Ames test and comet assay of first sample were carried out at concentrations of 100, 50, 25, 12.5, 6.25 % v/v effluent with distilled water. Chromium (Cr), lead (Pb), arsenic (As), and cadmium (Cd) were found in high concentrations as compared to WHO- and EPA-recommended maximum limits. Arsenic was found to be the most abundant metal and its maximum concentration was 0.8 mg.L(-1). GC-MS revealed the presence of lignocaine, digitoxin, trimethoprim, caffeine, and vitamin E in waste water. Dose-dependent decrease in mutagenic index was observed in both strains. Substantial increase in mutagenicity was observed for TA-100, when assay was done by incorporating an enzyme activation system, whereas a slight increase was detected for TA-102. In vitro comet assay of waste water exhibited decrease in damage index and percentage fragmentation with the increase in dilution of waste water. Tail length also decreased with an increase in the dilution factor of waste water. These findings suggest that pharmaceutical waste water being a mix of different heavy metals and organic contaminants may have a potent mutagenic and genotoxic effect on exposed living organisms.

Cardiac tissue enriched factors serum response factor and GATA-4 are mutual coregulators

NASA Technical Reports Server (NTRS)

Belaguli, N. S.; Sepulveda, J. L.; Nigam, V.; Charron, F.; Nemer, M.; Schwartz, R. J.

2000-01-01

Combinatorial interaction among cardiac tissue-restricted enriched transcription factors may facilitate the expression of cardiac tissue-restricted genes. Here we show that the MADS box factor serum response factor (SRF) cooperates with the zinc finger protein GATA-4 to synergistically activate numerous myogenic and nonmyogenic serum response element (SRE)-dependent promoters in CV1 fibroblasts. In the absence of GATA binding sites, synergistic activation depends on binding of SRF to the proximal CArG box sequence in the cardiac and skeletal alpha-actin promoter. GATA-4's C-terminal activation domain is obligatory for synergistic coactivation with SRF, and its N-terminal domain and first zinc finger are inhibitory. SRF and GATA-4 physically associate both in vivo and in vitro through their MADS box and the second zinc finger domains as determined by protein A pullout assays and by in vivo one-hybrid transfection assays using Gal4 fusion proteins. Other cardiovascular tissue-restricted GATA factors, such as GATA-5 and GATA-6, were equivalent to GATA-4 in coactivating SRE-dependent targets. Thus, interaction between the MADS box and C4 zinc finger proteins, a novel regulatory paradigm, mediates activation of SRF-dependent gene expression.

F.E.A.S.T.

MedlinePlus

... 2017 New blog: "Wasting Away": a Channel 4 documentary more Calendar The upcoming calendar is currently empty. ... O. Box 1281| Warrenton VA 20188 USA Association Management Software Powered by YourMembership :: Legal

Assessment of hazardous wastes for genotoxicity

DOE Office of Scientific and Technical Information (OSTI.GOV)

DeMarini, D.M.; Houk, V.S.

1987-09-01

The authors have evaluated a group of short-term bioassays to identify those that may be suitable for screening large numbers of diverse hazardous industrial wastes for genotoxicity. Fifteen wastes (and dichloromethane extracts of these wastes) from a variety of manufacturing processes were tested for mutagenicity in Salmonella typhimurium strains TA98 and TA100 with and without Aroclor 1254-induced rat-liver S9. Ten of these wastes were fed by gavage to F-344 male rats, and the raw urines were assayed for mutagenicity in the presence of beta-glucuronidase in strain TA98 with S9. Six of these urines were extracted by C18/methanol elution, incubated withmore » beta-glucuronidase, and evaluated in strain TA98 with S9 and beta-glucuronidase. Fourteen of the wastes were examined for their ability to induce prophage lambda in Escherichia coli in a microsuspension assay. A second set of wastes, consisting of four industrial wastes, were evaluated in Salmonella and in a series of mammalian cell assays to measure mutagenicity, cytogenetic effects, and transformation.« less

Active and passive computed tomography mixed waste focus area final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Roberson, G P

1998-08-19

The Mixed Waste Focus Area (MWFA) Characterization Development Strategy delineates an approach to resolve technology deficiencies associated with the characterization of mixed wastes. The intent of this strategy is to ensure the availability of technologies to support the Department of Energy's (DOE) mixed waste low-level or transuranic (TRU) contaminated waste characterization management needs. To this end the MWFA has defined and coordinated characterization development programs to ensure that data and test results necessary to evaluate the utility of non-destructive assay technologies are available to meet site contact handled waste management schedules. Requirements used as technology development project benchmarks are basedmore » in the National TRU Program Quality Assurance Program Plan. These requirements include the ability to determine total bias and total measurement uncertainty. These parameters must be completely evaluated for waste types to be processed through a given nondestructive waste assay system constituting the foundation of activities undertaken in technology development projects. Once development and testing activities have been completed, Innovative Technology Summary Reports are generated to provide results and conclusions to support EM-30, -40, or -60 end user/customer technology selection. The Active and Passive Computed Tomography non-destructive assay system is one of the technologies selected for development by the MWFA. Lawrence Livermore National Laboratory's (LLNL) is developing the Active and Passive Computed Tomography (A&PCT) nondestructive assay (NDA) technology to identify and accurately quantify all detectable radioisotopes in closed containers of waste. This technology will be applicable to all types of waste regardless of .their classification; low level, transuranic or provide results and conclusions to support EM-30, -40, or -60 end user/customer technology selection. The Active and Passive Computed Tomography non-destructive assay system is one of the technologies selected for development by the MWFA. Lawrence Livermore National Laboratory's (LLNL) is developing the Active and Passive Computed Tomography (A&PCT) nondestructive assay (NDA) technology to identify and accurately quantify all detectable radioisotopes in closed containers of waste. This technology will be applicable to all types of waste regardless of .their classification; low level, transuranic or mixed, which contains radioactivity and hazardous organic species. The scope of our technology is to develop a non-invasive waste-drum scanner that employs the principles of computed tomography and gamma-ray spectral analysis to identify and quantify all of the detectable radioisotopes. Once this and other applicable technologies are developed, waste drums can be non- destructively and accurately characterized to satisfy repository and regulatory guidelines prior to disposal.« less

The effect of recycled plastics and cooking oil on coke quality.

PubMed

Lange, Liséte Celina; Ferreira, Alison Frederico Medeiros

2017-03-01

This study assessed the effects of adding plastics and waste vegetable oil on the quality of coke in the coking process, on a pilot scale. A typical composition of the main plastics found in municipal solid waste was prepared using 33% HDPE, 5% LDPE, 10% PP, 21% PET, 24.8% PS, 5.2% PVC, 1% cellulose and also a 0.5% waste vegetable oil was added. The wastes were added to the coal blends in the proportions of 1%, 2% and 3% for plastics and 0.5% for vegetable oil. Two types of experiments were performed. The first was carried out in a hearth heating furnace (HHF) at temperatures of up to 900°C for a 7 h period. The second was a box test, which consists of heating coal blends in 18L cans using a pilot coking oven, for approximately 20 h at temperatures between 1050 and 1100°C. The quality parameters used for the assessment were the CSR (coke strength after reaction), CRI (coke reactivity index), ash, volatile matter and sulfur in order to identify the effect of plastic and vegetable oil on coke quality. Results for CSR in the HHF averaged 52.3%, and 56.63% in box test trials. The CRI results ranged from 26.6% to 35.7%. Among the different percentages of plastics used, 3% plastic blends provided the most stable CSR results. The industrial furnaces work at temperatures between 1100 and 1350°C and time coking 21-24h, compared to the test conditions achieved in the HHF and pilot furnace with box test. It was concluded that the results of CSR and CRI are consistent with the tests confirming the feasibility of using plastic in the steelmaking process. Copyright © 2016 Elsevier Ltd. All rights reserved.

VITRIFICATION SYSTEM FOR THE TREATMENT OF PLUTONIUM-BEARING WASTE AT LOS ALAMOS NATIONAL LABORATORY

DOE Office of Scientific and Technical Information (OSTI.GOV)

R. NAKAOKA; G. VEAZEY; ET AL

2001-05-01

A glove box vitrification system is being fabricated to process aqueous evaporator bottom waste generated at the Plutonium Facility (TA-55) at Los Alamos National Laboratory (LANL). The system will be the first within the U.S. Department of Energy Complex to routinely convert Pu{sup 239}-bearing transuranic (TRU) waste to a glass matrix for eventual disposal at the Waste Isolation Pilot Plant (WIPP). Currently at LANL, this waste is solidified in Portland cement. Radionuclide loading in the cementation process is restricted by potential radiolytic degradation (expressed as a wattage limit), which has been imposed to prevent the accumulation of flammable concentrations ofmore » H{sub 2} within waste packages. Waste matrixes with a higher water content (e.g., cement) are assigned a lower permissible wattage limit to compensate for their potential higher generation of H{sub 2}. This significantly increases the number of waste packages that must be prepared and shipped, thus driving up the costs of waste handling and disposal. The glove box vitrification system that is under construction will address this limitation. Because the resultant glass matrix produced by the vitrification process is non-hydrogenous, no H{sub 2} can be radiolytically evolved, and drums could be loaded to the maximum allowable limit of 40 watts. In effect, the glass waste form shifts the limiting constraint for loading disposal drums from wattage to the criticality limit of 200 fissile gram equivalents, thus significantly reducing the number of drums generated from this waste stream. It is anticipated that the number of drums generated from treatment of evaporator bottoms will be reduced by a factor of 4 annually when the vitrification system is operational. The system is currently undergoing non-radioactive operability testing, and will be fully operational in the year 2003.« less

Installation Restoration Program. Phase 1. Records Search, England AFB, Louisiana

DTIC Science & Technology

1983-05-01

compound shown on Figure 4.3. No herbicides, expired DDT or other pesticides were stored at this site. Some battery acid was stored in plastic boxes...Union Carbide Corporation, Chemicals and Plastics Divi- sion, Environomental Engineering Department. As a pro- cess/project engineer performed...paper mill waste treatment facility. Project Manager on Solid and Hazardous Waste study for a diverse chemicals and plastics production facility

Dynamics of TBP binding to the TATA box

NASA Astrophysics Data System (ADS)

Schluesche, Peter; Heiss, Gregor; Meisterernst, Michael; Lamb, Don C.

2008-02-01

Gene expression is highly controlled and regulated in living cells. One of the first steps in gene transcription is recognition of the promoter site by the TATA box Binding Protein (TBP). TBP recruits other transcriptions factors and eventually the RNA polymerase II to transcribe the DNA in mRNA. We developed a single pair Förster Resonance Energy Transfer (spFRET) assay to investigate the mechanism of gene regulation. Here, we apply this assay to investigate the initial binding process of TBP to the adenovirus major late (AdML) promoter site. From the spFRET measurements, we were able to identify two conformations of the TBP-DNA complex that correspond to TBP bound in the correct and the opposite orientation. Increased incubation times or the presence of the transcription factor TFIIA improved the alignment of TBP on the promoter site. Binding of TBP to the TATA box shows a rich dynamics with abrupt transitions between multiple FRET states. A frame-wise histogram analysis revealed the presence of at least six discrete states, showing that TBP binding is more complicated than previously thought. Hence, the spFRET assay is very sensitive to the conformation of the TBP-DNA complex and is very promising tool for investigating the pathway of TBP binding in detail.

A simple approach to the generation of heterologous competitive internal controls for real-time PCR assays on the LightCycler.

PubMed

Stöcher, Markus; Leb, Victoria; Hölzl, Gabriele; Berg, Jörg

2002-12-01

The real-time PCR technology allows convenient detection and quantification of virus derived DNA. This approach is used in many PCR based assays in clinical laboratories. Detection and quantification of virus derived DNA is usually performed against external controls or external standards. Thus, adequacy within a clinical sample is not monitored for. This can be achieved using internal controls that are co-amplified with the specific target within the same reaction vessel. We describe a convenient way to prepare heterologous internal controls as competitors for real-time PCR based assays. The internal controls were devised as competitors in real-time PCR, e.g. LightCycler-PCR. The bacterial neomycin phosphotransferase gene (neo) was used as source for heterologous DNA. Within the neo gene a box was chosen containing sequences for four differently spaced forward primers, one reverse primer, and a pair of neo specific hybridization probes. Pairs of primers were constructed to compose of virus-specific primer sequences and neo box specific primer sequences. Using those composite primers in conventional preparative PCR four types of internal controls were amplified from the neo box and subsequently cloned. A panel of the four differently sized internal controls was generated and tested by LightCycler PCR using their virus-specific primers. All four different PCR products were detected with the single pair of neo specific FRET-hybridization probes. The presented approach to generate competitive internal controls for use in LightCycler PCR assays proved convenient und rapid. The obtained internal controls match most PCR product sizes used in clinical routine molecular assays and will assist to discriminate true from false negative results.

Hazardous Waste Minimization and Treatment Opportunities in the Eighth U.S. Army and the U.S. Army, Japan

DTIC Science & Technology

1991-04-01

II), vapor degreasing unit using 1,1,1-trichloroethane, steam part cleaner using caustic soda, carburetor cleaning using cleaning compounds. * Bldg No...56 gal 7(" Dental clinic Hypo solution 15 gal Spent amalgam 8 lb 7 04 Medical x-ray H-yivo solution 240 gal 7(W Pathology lab Acetone and alcohols... spent millions of dollars to return PCP ammunition boxes to CONUS for disposal. However, a PCP ammunition box has never been hazardous material or

Waste from grocery stores

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lieb, K.

1993-11-01

The Community Recycling Center, Inc., (CRC, Champaign, Ill.), last year conducted a two-week audit of waste generated at two area grocery stores. The stores surveyed are part of a 10-store chain. For two of the Kirby Foods Stores, old corrugated containers (OCC) accounted for 39-45% of all waste. The summary drew correlations between the amount of OCC and the sum of food and garbage waste. The study suggested that one can reasonably estimate volumes of waste based on the amount of OCC because most things come in a box. Auditors set up a series of containers to make the collectionmore » process straightforward. Every day the containers were taken to local recycling centers and weighed. Approximate waste breakdowns for the two stores were as follows: 45% OCC; 35% food waste; 20% nonrecyclable or noncompostable items; and 10% other.« less

Role of the Box C/D Motif in Localization of Small Nucleolar RNAs to Coiled Bodies and Nucleoli

PubMed Central

Narayanan, Aarthi; Speckmann, Wayne; Terns, Rebecca; Terns, Michael P.

1999-01-01

Small nucleolar RNAs (snoRNAs) are a large family of eukaryotic RNAs that function within the nucleolus in the biogenesis of ribosomes. One major class of snoRNAs is the box C/D snoRNAs named for their conserved box C and box D sequence elements. We have investigated the involvement of cis-acting sequences and intranuclear structures in the localization of box C/D snoRNAs to the nucleolus by assaying the intranuclear distribution of fluorescently labeled U3, U8, and U14 snoRNAs injected into Xenopus oocyte nuclei. Analysis of an extensive panel of U3 RNA variants showed that the box C/D motif, comprised of box C′, box D, and the 3′ terminal stem of U3, is necessary and sufficient for the nucleolar localization of U3 snoRNA. Disruption of the elements of the box C/D motif of U8 and U14 snoRNAs also prevented nucleolar localization, indicating that all box C/D snoRNAs use a common nucleolar-targeting mechanism. Finally, we found that wild-type box C/D snoRNAs transiently associate with coiled bodies before they localize to nucleoli and that variant RNAs that lack an intact box C/D motif are detained within coiled bodies. These results suggest that coiled bodies play a role in the biogenesis and/or intranuclear transport of box C/D snoRNAs. PMID:10397754

Solving the Pallet Loading Problem

DTIC Science & Technology

2008-01-01

solution M = N(X,Y,a,b) there must be at least one unit column with zero waste because W(M,X,Y,a,b) < X + a and therefore W(M 0,X + a, Y,a,b) < X + a for M0...P M + By. Any such unit column with zero waste must be covered with H-boxes because this corresponds to the only perfect Y-partition. If there is a

The bZIP transcription factor HY5 interacts with the promoter of the monoterpene synthase gene QH6 in modulating its rhythmic expression.

PubMed

Zhou, Fei; Sun, Tian-Hu; Zhao, Lei; Pan, Xi-Wu; Lu, Shan

2015-01-01

The Artemisia annua L. β-pinene synthase QH6 was previously determined to be circadian-regulated at the transcriptional level, showing a rhythmic fluctuation of steady-state transcript abundances. Here we isolated both the genomic sequence and upstream promoter region of QH6. Different regulatory elements, such as G-box (TGACACGTGGCA, -421 bp from the translation initiation site) which might have effects on rhythmic gene expression, were found. Using the yeast one-hybrid and electrophoretic mobility shift assay (EMSA), we confirmed that the bZIP transcription factor HY5 binds to this motif of QH6. Studies with promoter truncations before and after this motif suggested that this G-box was important for the diurnal fluctuation of the transgenic β-glucuronidase gene (GUS) transcript abundance in Arabidopsis thaliana. GUS gene driven by the promoter region immediately after G-box showed an arrhythmic expression in both light/dark (LD) and constant dark (DD) conditions, whereas the control with G-box retained its fluctuation in both LD and DD. We further transformed A. thaliana with the luciferase gene (LUC) driven by an 1400 bp fragment upstream QH6 with its G-box intact or mutated, respectively. The luciferase activity assay showed that a peak in the early morning disappeared in the mutant. Gene expression analysis also demonstrated that the rhythmic expression of LUC was abolished in the hy5-1 mutant.

Elucidating the evolutionary conserved DNA-binding specificities of WRKY transcription factors by molecular dynamics and in vitro binding assays

PubMed Central

Brand, Luise H.; Fischer, Nina M.; Harter, Klaus; Kohlbacher, Oliver; Wanke, Dierk

2013-01-01

WRKY transcription factors constitute a large protein family in plants that is involved in the regulation of developmental processes and responses to biotic or abiotic stimuli. The question arises how stimulus-specific responses are mediated given that the highly conserved WRKY DNA-binding domain (DBD) exclusively recognizes the ‘TTGACY’ W-box consensus. We speculated that the W-box consensus might be more degenerate and yet undetected differences in the W-box consensus of WRKYs of different evolutionary descent exist. The phylogenetic analysis of WRKY DBDs suggests that they evolved from an ancestral group IIc-like WRKY early in the eukaryote lineage. A direct descent of group IIc WRKYs supports a monophyletic origin of all other group II and III WRKYs from group I by loss of an N-terminal DBD. Group I WRKYs are of paraphyletic descent and evolved multiple times independently. By homology modeling, molecular dynamics simulations and in vitro DNA–protein interaction-enzyme-linked immunosorbent assay with AtWRKY50 (IIc), AtWRKY33 (I) and AtWRKY11 (IId) DBDs, we revealed differences in DNA-binding specificities. Our data imply that other components are essentially required besides the W-box-specific binding to DNA to facilitate a stimulus-specific WRKY function. PMID:23975197

The Twist Box Domain is Required for Twist1-induced Prostate Cancer Metastasis

PubMed Central

Gajula, Rajendra P.; Chettiar, Sivarajan T.; Williams, Russell D.; Thiyagarajan, Saravanan; Kato, Yoshinori; Aziz, Khaled; Wang, Ruoqi; Gandhi, Nishant; Wild, Aaron T.; Vesuna, Farhad; Ma, Jinfang; Salih, Tarek; Cades, Jessica; Fertig, Elana; Biswal, Shyam; Burns, Timothy F.; Chung, Christine H.; Rudin, Charles M.; Herman, Joseph M.; Hales, Russell K.; Raman, Venu; An, Steven S.; Tran, Phuoc T.

2013-01-01

Twist1, a basic helix-loop-helix transcription factor, plays a key role during development and is a master regulator of the epithelial-mesenchymal transition (EMT) that promotes cancer metastasis. Structure-function relationships of Twist1 to cancer-related phenotypes are underappreciated, so we studied the requirement of the conserved Twist box domain for metastatic phenotypes in prostate cancer (PCa). Evidence suggests that Twist1 is overexpressed in clinical specimens and correlated with aggressive/metastatic disease. Therefore, we examined a transactivation mutant, Twist1-F191G, in PCa cells using in vitro assays which mimic various stages of metastasis. Twist1 overexpression led to elevated cytoskeletal stiffness and cell traction forces at the migratory edge of cells based on biophysical single-cell measurements. Twist1 conferred additional cellular properties associated with cancer cell metastasis including increased migration, invasion, anoikis resistance, and anchorage-independent growth. The Twist box mutant was defective for these Twist1 phenotypes in vitro. Importantly, we observed a high frequency of Twist1-induced metastatic lung tumors and extra-thoracic metastases in vivo using the experimental lung metastasis assay. The Twist box was required for PCa cells to colonize metastatic lung lesions and extra-thoracic metastases. Comparative genomic profiling revealed transcriptional programs directed by the Twist box that were associated with cancer progression, such as Hoxa9. Mechanistically, Twist1 bound to the Hoxa9 promoter and positively regulated Hoxa9 expression in PCa cells. Finally, Hoxa9 was important for Twist1-induced cellular phenotypes associated with metastasis. These data suggest that the Twist box domain is required for Twist1 transcriptional programs and PCa metastasis. PMID:23982216

Toxicity assessment of carbon black waste: A by-product from oil refineries.

PubMed

Zhen, Xu; Ng, Wei Cheng; Fendy; Tong, Yen Wah; Dai, Yanjun; Neoh, Koon Gee; Wang, Chi-Hwa

2017-01-05

In Singapore, approximately 30t/day of carbon-based solid waste are produced from petrochemical processes. This carbon black waste has been shown to possess physical properties that are characteristic of a good adsorbent such as high external surface area. Therefore, there is a growing interest to reutilize and process this carbon black waste into secondary materials such as adsorbents. However, the carbon black waste obtained from petrochemical industries may contain heavy metals that are hazardous to human health and the environment, hence restricting its full potential for re-utilization. Therefore, it is important to examine the possible toxicity effects and toxicity mechanism of carbon black waste on human health. In this study, inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis showed that the heavy metals, vanadium (V), molybdenum (Mo) and nickel (Ni), were present in the carbon black waste in high concentrations. Three human cell lines (HepG2 cells, MRC-5 cells and MDA-MB-231 cells) were used to investigate the toxicity of carbon black waste extract in a variety of in vitro assays. Results from MTS assays indicated that carbon black waste extract decreased the viability of all three cell lines in a dose and time-dependent manner. Observations from confocal microscopy further confirmed this phenomenon. Flow cytometry assay also showed that carbon black waste extract induced apoptosis of human cell lines, and the level of apoptosis increased with increasing waste concentration. Results from reactive oxygen species (ROS) assay indicated that carbon black waste extract induced oxidative stress by increasing intracellular ROS generation in these three human cell lines. Moreover, induction of oxidative damage in these cells was also observed through the alteration of glutathione (GSH) and superoxide dismutase (SOD) activities. Last but not least, by treating the cells with V-spiked solution of concentration equivalent to that found in the carbon black waste extract, V was identified as the main culprit for the high toxicity of carbon black waste extract. These findings could potentially provide insight into the hazards of carbon black waste extract and its toxicity mechanism on human cell lines. Copyright © 2016 Elsevier B.V. All rights reserved.

Polyhydroxybutyrate (PHB) Synthesis by Spirulina sp. LEB 18 Using Biopolymer Extraction Waste.

PubMed

da Silva, Cleber Klasener; Costa, Jorge Alberto Vieira; de Morais, Michele Greque

2018-01-20

The reuse of waste as well as the production of biodegradable compounds has for years been the object of studies and of global interest as a way to reduce the environmental impact generated by unsustainable exploratory processes. The conversion of linear processes into cyclical processes has environmental and economic advantages, reducing waste deposition and reducing costs. The objective of this work was to use biopolymer extraction waste in the cultivation of Spirulina sp. LEB 18, for the cyclic process of polyhydroxybutyrate (PHB) synthesis. Concentrations of 10, 15, 20, 25, and 30% (v/v) of biopolymer extraction waste were tested. For comparison, two assays were used without addition of waste, Zarrouk (SZ) and modified Zarrouk (ZM), with reduction of nitrogen. The assays were carried out in triplicate and evaluated for the production of microalgal biomass and PHB. The tests with addition of waste presented a biomass production statistically equal to ZM (0.79 g L -1 ) (p < 0.1). The production of PHB in the assay containing 25% of waste was higher when compared to the other cultivations, obtaining 10.6% (w/w) of biopolymer. From the results obtained, it is affirmed that the use of PHB extraction waste in the microalgal cultivation, aiming at the synthesis of biopolymers, can occur in a cyclic process, reducing process costs and the deposition of waste, thus favoring the preservation of the environment.

40 CFR 63.8266 - What definitions apply to this subpart?

Code of Federal Regulations, 2013 CFR

2013-07-01

... all vessels, piping, and equipment that evacuate the head space of each mercury cell end box (and...-containing wastes, further classified as either debris or nondebris (i.e., cell room sludges and carbon from...

40 CFR 63.8266 - What definitions apply to this subpart?

Code of Federal Regulations, 2012 CFR

2012-07-01

... all vessels, piping, and equipment that evacuate the head space of each mercury cell end box (and...-containing wastes, further classified as either debris or nondebris (i.e., cell room sludges and carbon from...

The plant G box promoter sequence activates transcription in Saccharomyces cerevisiae and is bound in vitro by a yeast activity similar to GBF, the plant G box binding factor.

PubMed Central

Donald, R G; Schindler, U; Batschauer, A; Cashmore, A R

1990-01-01

G box and I box sequences of the Arabidopsis thaliana ribulose-bisphosphate-1,5-carboxylase small subunit (RBCS) promoter are required for expression mediated by the Arabidopsis rbcS-1A promoter in transgenic tobacco plants and are bound in vitro by factors from plant nuclear extracts termed GBF and GA-1, respectively. We show here that a -390 to -60 rbcS-1A promoter fragment containing the G box and two I boxes activates transcription from a truncated iso-1-cytochrome c (CYC1) gene promoter in Saccharomyces cerevisiae. Mutagenesis of either the rbcS-1A G box or both I box sequences eliminated the expression mediated by this fragment. When polymerized, I box oligonucleotides were also capable of enhancing expression from the truncated CYC1 promoter. Single-copy G box sequences from the Arabidopsis rbcS-1A, Arabidopsis Adh and tomato rbcS-3A promoters were more potent activators and were used in mobility shift assays to identify a DNA binding activity in yeast functionally similar to GBF. In methylation interference experiments, the binding specificity of the yeast protein was indistinguishable from that obtained with plant nuclear extracts. Images Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:2161333

Deciphering the molecular mechanisms underlying the binding of the TWIST1/E12 complex to regulatory E-box sequences

PubMed Central

Bouard, Charlotte; Terreux, Raphael; Honorat, Mylène; Manship, Brigitte; Ansieau, Stéphane; Vigneron, Arnaud M.; Puisieux, Alain; Payen, Léa

2016-01-01

Abstract The TWIST1 bHLH transcription factor controls embryonic development and cancer processes. Although molecular and genetic analyses have provided a wealth of data on the role of bHLH transcription factors, very little is known on the molecular mechanisms underlying their binding affinity to the E-box sequence of the promoter. Here, we used an in silico model of the TWIST1/E12 (TE) heterocomplex and performed molecular dynamics (MD) simulations of its binding to specific (TE-box) and modified E-box sequences. We focused on (i) active E-box and inactive E-box sequences, on (ii) modified active E-box sequences, as well as on (iii) two box sequences with modified adjacent bases the AT- and TA-boxes. Our in silico models were supported by functional in vitro binding assays. This exploration highlighted the predominant role of protein side-chain residues, close to the heart of the complex, at anchoring the dimer to DNA sequences, and unveiled a shift towards adjacent ((-1) and (-1*)) bases and conserved bases of modified E-box sequences. In conclusion, our study provides proof of the predictive value of these MD simulations, which may contribute to the characterization of specific inhibitors by docking approaches, and their use in pharmacological therapies by blocking the tumoral TWIST1/E12 function in cancers. PMID:27151200

Characterization of temporal variations in landfill gas components inside an open solid waste dump site in Sri Lanka.

PubMed

Nagamori, Masanao; Mowjood, M I M; Watanabe, Youichi; Isobe, Yugo; Ishigaki, Tomonori; Kawamoto, Ken

2016-12-01

A long-term monitoring of composition of landfill gases in the region with high rainfall was conducted using an argon assay in order to discuss air intrusion into the dump site. Gas samples were taken from vertical gas monitoring pipes installed along transects at two sections (called new and old) of an abandoned waste dump site in Sri Lanka. N 2 O concentrations varied especially widely, by more than three orders of magnitude (0.046-140 ppmv). The nitrogen/argon ratio of landfill gas was normally higher than that of fresh air, implying that denitrification occurred in the dump site. Argon assays indicate that both N 2 and N 2 O production occurred inside waste and more significantly in the old section. The Ar assay would help for evaluations of N 2 O emission in developing countries. A long-term monitoring of composition of landfill gases in the region with high rainfall was conducted using an argon assay in order to discuss air intrusion into the dump site. Argon assays indicate that both N 2 and N 2 O production occurred inside waste and more significantly in the old section.

Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study on Xenopus laevis Larvae Assay with Biologically Active Substances Present in Livestock Wastes

PubMed Central

Martini, Federica; Tarazona, José V.; Pablos, M. Victoria

2012-01-01

Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians. PMID:22629159

Screening of the MMV and GSK open access chemical boxes using a viability assay developed against the kinetoplastid Crithidia fasciculata.

PubMed

Kipandula, Wakisa; Young, Simon A; MacNeill, Stuart A; Smith, Terry K

2018-06-01

Diseases caused by the pathogenic kinetoplastids continue to incapacitate and kill hundreds of thousands of people annually throughout the tropics and sub-tropics. Unfortunately, in the countries where these neglected diseases occur, financial obstacles to drug discovery and technical limitations associated with biochemical studies impede the development of new, safe, easy to administer and effective drugs. Here we report the development and optimisation of a Crithidia fasciculata resazurin viability assay, which is subsequently used for screening and identification of anti-crithidial compounds in the MMV and GSK open access chemical boxes. The screening assay had an average Z' factor of 0.7 and tolerated a maximum dimethyl sulfoxide concentration of up to 0.5%. We identified from multiple chemical boxes two compound series exhibiting nanomolar potency against C. fasciculata, one centred around a 5-nitrofuran-2-yl scaffold, a well-known moiety in several existing anti-infectives, and another involving a 2-(pyridin-2-yl) pyrimidin-4-amine scaffold which seems to have pan-kinetoplastid activity. This work facilitates the future use of C. fasciculata as a non-pathogenic and inexpensive biological resource to identify mode of action/protein target(s) of potentially pan-trypanocidal potent compounds. This knowledge will aid in the development of new treatments for African sleeping sickness, Chagas disease and leishmaniasis. Copyright © 2018 Elsevier B.V. All rights reserved.

Development and validation of quantitative PCR for detection of Terrapene herpesvirus 1 utilizing free-ranging eastern box turtles (Terrapene carolina carolina).

PubMed

Kane, Lauren P; Bunick, David; Abd-Eldaim, Mohamed; Dzhaman, Elena; Allender, Matthew C

2016-06-01

Diseases that affect the upper respiratory tract (URT) in chelonians have been well described as a significant contributor of morbidity and mortality. Specifically, herpesviruses are common pathogens in captive chelonians worldwide, but their importance on free-ranging populations is less well known. Historical methods for the diagnosis of herpesvirus infections include virus isolation and conventional PCR. Real-time PCR has become an essential tool for detection and quantitation of many pathogens, but has not yet been developed for herpesviruses in box turtles. Two quantitative real-time TaqMan PCR assays, TerHV58 and TerHV64, were developed targeting the DNA polymerase gene of Terrapene herpesvirus 1 (TerHV1). The assay detected a viral DNA segment cloned within a plasmid with 10-fold serial dilutions from 1.04 × 10(7) to 1.04 × 10(1) viral copies per reaction. Even though both primers had acceptable levels of efficiency and variation, TerHV58 was utilized to test clinical samples based on less variation and increased efficiency. This assay detected as few as 10 viral copies per reaction and should be utilized in free-ranging and captive box turtles to aid in the characterization of the epidemiology of this disease. Copyright © 2016 Elsevier B.V. All rights reserved.

Negative modulation of the chicken infectious anemia virus promoter by COUP-TF1 and an E box-like element at the transcription start site binding deltaEF1.

PubMed

Miller, Myrna M; Jarosinski, Keith W; Schat, Karel A

2008-12-01

Expression of enhanced green fluorescent protein (EGFP) under control of the promoter-enhancer of chicken infectious anemia virus (CAV) is increased in an oestrogen receptor-enhanced cell line when treated with oestrogen and the promoter-enhancer binds unidentified proteins that recognize a consensus oestrogen response element (ERE). Co-transfection assays with the CAV promoter and the nuclear receptor chicken ovalbumin upstream promoter transcription factor 1 (COUP-TF1) showed that expression of EGFP was decreased by 50 to 60 % in DF-1 and LMH cells. The CAV promoter that included sequences at and downstream of the transcription start point had less expression than a short promoter construct. Mutation of a putative E box at this site restored expression levels. Electromobility shift assays showed that the transcription regulator delta-EF1 (deltaEF1) binds to this E box region. These findings indicate that the CAV promoter activity can be affected directly or indirectly by COUP-TF1 and deltaEF1.

Design and adaptation of a novel supercritical extraction facility for operation in a glove box for recovery of radioactive elements

NASA Astrophysics Data System (ADS)

Kumar, V. Suresh; Kumar, R.; Sivaraman, N.; Ravisankar, G.; Vasudeva Rao, P. R.

2010-09-01

The design and development of a novel supercritical extraction experimental facility adapted for safe operation in a glove box for the recovery of radioactive elements from waste is described. The apparatus incorporates a high pressure extraction vessel, reciprocating pumps for delivering supercritical fluid and reagent, a back pressure regulator, and a collection chamber. All these components of the system have been specially designed for glove box adaptation and made modular to facilitate their replacement. Confinement of these materials must be ensured in a glove box to protect the operator and prevent contamination to the work area. Since handling of radioactive materials under high pressure (30 MPa) and temperature (up to 333 K) is involved in this process, the apparatus needs elaborate safety features in the design of the equipment, as well as modification of a standard glove box to accommodate the system. As a special safety feature to contain accidental leakage of carbon dioxide from the extraction vessel, a safety vessel has been specially designed and placed inside the glove box. The extraction vessel was enclosed in the safety vessel. The safety vessel was also incorporated with pressure sensing and controlling device.

Mitigated Transfer Line Leaks that Result in Surface Pools and Spray Leaks into Pits

DOE Office of Scientific and Technical Information (OSTI.GOV)

HEY, B.E.

This analysis provides radiological and toxicological consequence calculations for postulated mitigated leaks during transfers of six waste compositions. Leaks in Cleanout Boxes equipped with supplemental covers and leaks in pits are analyzed.

Solar disinfection of infectious biomedical waste: a new approach for developing countries.

PubMed

Chitnis, V; Chitnis, S; Patil, S; Chitnis, D

2003-10-18

Poor developing countries cannot afford expensive technologies such as incineration for management of infectious biomedical waste. We assessed solar heating as an alternative technology. We immersed simulated infectious waste with added challenge bacteria in water in a box-type solar cooker, which was left in the sun for 6 h. In 24 sets of observations, the amount of viable bacteria was reduced by about 7 log. We also tested infectious medical waste with a heavy load of bacteria (10(8)-10(9)/g) from our hospital's burn unit for solar heat disinfection in 20 experiments. Our results showed a similar 7 log reduction in the amount of viable bacteria. Solar heating thus seems to be a cheap method to disinfect infectious medical waste in less economically developed countries.

Structure-Function Relationships in Human Testis-determining Factor SRY

PubMed Central

Racca, Joseph D.; Chen, Yen-Shan; Maloy, James D.; Wickramasinghe, Nalinda; Phillips, Nelson B.; Weiss, Michael A.

2014-01-01

Human testis determination is initiated by SRY, a Y-encoded architectural transcription factor. Mutations in SRY cause 46 XY gonadal dysgenesis with female somatic phenotype (Swyer syndrome) and confer a high risk of malignancy (gonadoblastoma). Such mutations cluster in the SRY high mobility group (HMG) box, a conserved motif of specific DNA binding and bending. To explore structure-function relationships, we constructed all possible substitutions at a site of clinical mutation (W70L). Our studies thus focused on a core aromatic residue (position 15 of the consensus HMG box) that is invariant among SRY-related HMG box transcription factors (the SOX family) and conserved as aromatic (Phe or Tyr) among other sequence-specific boxes. In a yeast one-hybrid system sensitive to specific SRY-DNA binding, the variant domains exhibited reduced (Phe and Tyr) or absent activity (the remaining 17 substitutions). Representative nonpolar variants with partial or absent activity (Tyr, Phe, Leu, and Ala in order of decreasing side-chain volume) were chosen for study in vitro and in mammalian cell culture. The clinical mutation (Leu) was found to markedly impair multiple biochemical and cellular activities as respectively probed through the following: (i) in vitro assays of specific DNA binding and protein stability, and (ii) cell culture-based assays of proteosomal degradation, nuclear import, enhancer DNA occupancy, and SRY-dependent transcriptional activation. Surprisingly, however, DNA bending is robust to this or the related Ala substitution that profoundly impairs box stability. Together, our findings demonstrate that the folding, trafficking, and gene-regulatory function of SRY requires an invariant aromatic “buttress” beneath its specific DNA-bending surface. PMID:25258310

TECHNICAL ASSESSMENT OF BULK VITRIFICATION PROCESS & PRODUCT FOR TANK WASTE TREATMENT AT THE DEPARTMENT OF ENERGY HANFORD SITE

DOE Office of Scientific and Technical Information (OSTI.GOV)

SCHAUS, P.S.

At the U.S. Department of Energy (DOE) Hanford Site, the Waste Treatment Plant (WTP) is being constructed to immobilize both high-level waste (IUW) for disposal in a national repository and low-activity waste (LAW) for onsite, near-surface disposal. The schedule-controlling step for the WTP Project is vitrification of the large volume of LAW, current capacity of the WTP (as planned) would require 50 years to treat the Hanford tank waste, if the entire LAW volume were to be processed through the WTP. To reduce the time and cost for treatment of Hanford Tank Waste, and as required by the Tank Wastemore » Remediation System Environmental Impact Statement Record of Decision and the Hanford Federal Facility Consent Agreement (Tn-Party Agreement), DOE plans to supplement the LAW treatment capacity of the WTP. Since 2002, DOE, in cooperation with the Environmental Protection Agency and State of Washington Department of Ecology has been evaluating technologies that could provide safe and effective supplemental treatment of LAW. Current efforts at Hanford are intended to provide additional information to aid a joint agency decision on which technology will be used to supplement the WTP. A Research, Development and Demonstration permit has been issued by the State of Washington to build and (for a limited time) operate a Demonstration Bulk Vitrification System (DBVS) facility to provide information for the decision on a supplemental treatment technology for up to 50% of the LAW. In the Bulk Vitrification (BV) process, LAW, soil, and glass-forming chemicals are mixed, dried, and placed in a refractory-lined box, Electric current, supplied through two graphite electrodes in the box, melts the waste feed, producing a durable glass waste-form. Although recent modifications to the process have resulted in significant improvements, there are continuing technical concerns.« less

Mobile site safety review for the transuranic (TRU) waste characterization program

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1996-11-01

This Safety Review Document (SRD) applies to the Active/Passive Neutron Examination and Assay (APNEA) system installed on a Lockheed Martin Specialty Components, Inc., (Specialty Components) trailer. The APNEA is designed to perform nuclear waste drum assay. The purpose of this document is to describe the safety features of the APNEA system.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Weaver, Brian Phillip

The purpose of this document is to describe the statistical modeling effort for gas concentrations in WIPP storage containers. The concentration (in ppm) of CO 2 in the headspace volume of standard waste box (SWB) 68685 is shown. A Bayesian approach and an adaptive Metropolis-Hastings algorithm were used.

Screening complex hazardous wastes for mutagenic activity using a modified version of the TLC/Salmonella assay.

PubMed

Houk, V S; Claxton, L D

1986-03-01

10 complex hazardous wastes were tested for mutagenic activity using a modified version of the TLC/Salmonella assay developed by Bjørseth et al. (1982). This fractionation/bioassay scheme couples thin-layer chromatography (TLC) with the Salmonella/mammalian-microsome (Ames) assay for the detection of mutagenic constituents in complex mixtures. Crude (unadulterated) hazardous wastes and selected hazardous waste extracts were fractionated on commercially available cellulose TLC plates. Mutagenicity testing was performed in situ by applying a single overlay of minimal growth agar, tester strain TA98 or TA100, and the optional metabolic activation system directly onto the developed chromatogram. A mutagenic effect was indicated either by the appearance of localized clusters of revertant colonies or by an increase in total revertant growth vis-à-vis control plates. 7 of 10 hazardous wastes (including tars, emulsions, sludges, and spent acids and caustics) demonstrated mutagenic activity when tested by this method. To assess the sensitivity of the modified TLC/Salmonella assay, 14 Salmonella mutagens from a wide range of chemical classes and polarities were tested. Selected compounds included heterocyclics, aromatic amines, alkylating agents, antitumor agents, a nitrosamine and a nitroaromatic. 11 of the 14 mutagens were positive in this test system. The 3 compounds refractory to analysis included a polycyclic aromatic hydrocarbon and two volatiles.

Forkhead box K2 inhibits the proliferation, migration, and invasion of human glioma cells and predicts a favorable prognosis.

PubMed

Wang, Bo; Zhang, XueBin; Wang, Wei; Zhu, ZhiZhong; Tang, Fan; Wang, Dong; Liu, Xi; Zhuang, Hao; Yan, XiaoLing

2018-01-01

Forkhead box K2 (FOXK2) is a member of the forkhead box family of transcription factors. Recently, researchers discovered that overexpression of FOXK2 inhibits the proliferation and metastasis of breast cancer, non-small cell lung cancer, and colorectal cancer, and is related to the clinical prognosis. However, in hepatocellular carcinoma, FOXK2 results in the opposite phenotypes. Currently, the contribution of FOXK2 to glioma pathogenesis is not clear. We evaluated the expression of FOXK2 in 151 glioma patients using immunohistochemistry assays. The associations among the expression of FOXK2, clinicopathological parameters, and the prognosis of glioma patients were statistically analyzed. We downregulated and upregulated the level of FOXK2 in glioma cells by transfections with small interfering RNA and plasmids. Then, we investigated the effects on tumor cell behavior in vitro by Cell Counting Kit-8 assays, colony-formation assay, transwell assay, and the epithelial-to-mesenchymal transition (EMT) biomarker levels. The clinical data showed that expression of FOXK2 gradually decreased with increasing World Health Organization (WHO) grades and a low level of FOXK2 indicates a poor prognosis. FOXK2 expression is negatively correlated with Ki67 expression and the WHO degree but is not correlated with other clinicopathological parameters, including sex, age, Karnofsky Performance Status, tumor diameter, O -6-methylguanine-DNA methyltransferase, and glutathione S -transferase pi. FOXK2 knockdown enhances glioma cell proliferation, migration, invasion, and EMT process, and, in contrast, FOXK2 overexpression inhibits glioma cell proliferation, migration, invasion, and the EMT process. Expression of FOXK2 gradually decreases with increasing WHO grades. FOXK2 inhibits tumor proliferation, migration, and invasion. FOXK2 is a critical mediator of the EMT process.

Facile Synthesis of Three-Dimensional Heteroatom-Doped and Hierarchical Egg-Box-Like Carbons Derived from Moringa oleifera Branches for High-Performance Supercapacitors.

PubMed

Cai, Yijin; Luo, Ying; Xiao, Yong; Zhao, Xiao; Liang, Yeru; Hu, Hang; Dong, Hanwu; Sun, Luyi; Liu, Yingliang; Zheng, Mingtao

2016-12-07

In this paper, we demonstrate that Moringa oleifera branches, a renewable biomass waste with abundant protein content, can be employed as novel precursor to synthesize three-dimensional heteroatom-doped and hierarchical egg-box-like carbons (HEBLCs) by a facile room-temperature pretreatment and direct pyrolysis process. The as-prepared HEBLCs possess unique egg-box-like frameworks, high surface area, and interconnected porosity as well as the doping of heteroatoms (oxygen and nitrogen), endowing its excellent electrochemical performances (superior capacity, high rate capability, and outstanding cycling stability). Therefore, the resultant HEBLC manifests a maximum specific capacitance of 355 F g -1 at current density of 0.5 A g -1 and remarkable rate performance. Moreover, 95% of capacitance retention of HEBLCs can be also achieved after 20 000 charge-discharge cycles at an extremely high current density (20 A g -1 ), indicating a prominent cycling stability. Furthermore, the as-assembled HEBLC//HEBLC symmetric supercapacitor displays a superior energy density of 20 Wh kg -1 in aqueous electrolyte and remarkable capacitance retention (95.6%) after 10 000 charge-discharge cycles. This work provides an environmentally friendly and reliable method to produce higher-valued carbon nanomaterials from renewable biomass wastes for energy storage applications.

Performance test results of noninvasive characterization of Resource Conservation and Recovery Act surrogate waste by prompt gamma neutron activation analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gehrke, R.J.; Streier, G.G.

1997-03-01

During FY-96, a performance test was carried out with funding from the Mixed Waste Focus Area (MWFA) of the Department of Energy (DOE) to determine the noninvasive elemental assay capabilities of commercial companies for Resource Conservation and Recovery Act (RCRA) metals present in 8-gal drums containing surrogate waste. Commercial companies were required to be experienced in the use of prompt gamma neutron activation analysis (PGNAA) techniques and to have a prototype assay system with which to conduct the test assays. Potential participants were identified through responses to a call for proposals advertised in the Commerce Business Daily and through personalmore » contacts. Six companies were originally identified. Two of these six were willing and able to participate in the performance test, as described in the test plan, with some subsidizing from the DOE MWFA. The tests were conducted with surrogate sludge waste because (1) a large volume of this type of waste awaits final disposition and (2) sludge tends to be somewhat homogeneous. The surrogate concentrations of the above RCRA metals ranged from {approximately} 300 ppm to {approximately} 20,000 ppm. The lower limit was chosen as an estimate of the expected sensitivity of detection required by noninvasive, pretreatment elemental assay systems to be of value for operational and compliance purposes and to still be achievable with state-of-the-art methods of analysis. The upper limit of {approximately} 20,000 ppm was chosen because it is the opinion of the author that assay above this concentration level is within current state-of-the-art methods for most RCRA constituents. This report is organized into three parts: Part 1, Test Plan to Evaluate the Technical Status of Noninvasive Elemental Assay Techniques for Hazardous Waste; Part 2, Participants` Results; and Part 3, Evaluation of and Comments on Participants` Results.« less

Dehydrogenase activity and quality of leachates in Technosols with gossan and sulfide materials from the São Domingos mine

NASA Astrophysics Data System (ADS)

Santos, Erika; Abreu, Manuela; Macías, Felipe; de Varennes, Amarílis

2014-05-01

Wastes produced by mining activity in São Domingos (Portuguese Iberian Pyrite Belt) were disposed over a large area. To speed up the ecological rehabilitation in this mine, an integrative strategy using different amendments+mine wastes was used to produce Technosols with enhanced soil functions. To evaluate the efficiency of these Technosols the dehydrogenase activity and chemical quality of leachates were monitored. Technosols were composed of different mine wastes (gossan and sulfide materials), collected at the São Domingos mine, and mixtures of amendments applied at 30 and 75 Mg/ha (rockwool+agriculture wastes+wastes from liquors distillation of strawberry tree fruits (Arbutus unedo L.) and/or carobs (Ceratonia siliqua L. fruits)). Three assays, under controlled conditions, were carried out: (1 and 2) Sulfide or gossan materials with/without amendments; (3) Sulfide wastes, with/without amendments, incubated during four months and then with application of an overlayer of gossan (~3 cm thick) with/without the same amendments. Dehydrogenase activity (DHA) and chemical characteristics of leachates (multielemental concentration, pH, and electric conductivity) were determined after four/seven/thirteen months of incubation. Sulfide wastes had more hazardous characteristics (pH~2 and total concentrations (g/kg) of Al (58.1), As (1.1), Cu (2.1), Fe (107.3), Pb (11.7), S (65.3) and Zn (1.1) than the gossan materials (pH=4.3; g/kg, Al: 24.8, As: 3.0, Cu: 0.2, Fe: 129, Pb: 9.2, S: 13.7, Zn: 0.04). Amendments application to gossan (assay 2) enhanced DHA in both sampling periods (µg TPF g dry weight 16 h-1, Control: 0,72-1,78; Amended treatments: 2.49-16.36 depending on mixture/application rate/sampling period). Greater application rates stimulated DHA (more than 1.5-fold with 75 Mg/ha). No differences were observed in DHA in the gossan layer with/without amendments (assay 3) suggesting a negative impact on gossan microrganisms from sulfide materials located below. In fact, for sulfide materials (assays 1 and 3), the amendments did not lead to increases in DHA (µg TPF g dry weight 16 h-1, Control: 0.87-2.72; Amended treatments: 0.23-2.11, depending on assay/sampling period/treatment). In assay 3, the DHA of each mine waste presented, in general, smaller enzymatic activity compared to the same treatments from the other assays. The amendments reduced the electrical condutivity (35-60 %) in the leachates from Technosols with sulfide materials but the pH of leachates increased in all cases between 0.5 and 1.5 units, depending on assay/treatment/sampling period.

Warehouse hazardous and toxic waste design in Karingau Balikpapan

NASA Astrophysics Data System (ADS)

Pratama, Bayu Rendy; Kencanawati, Martheana

2017-11-01

PT. Balikpapan Environmental Services (PT. BES) is company that having core business in Hazardous and Toxic Waste Management Services which consisting storage and transporter at Balikpapan. This research starting with data collection such as type of waste, quantity of waste, dimension area of existing building, waste packaging (Drum, IBC tank, Wooden Box, & Bulk Bag). Processing data that will be done are redesign for warehouse dimension and layout of position waste, specify of capacity, specify of quantity, type and detector placement, specify of quantity, type and fire extinguishers position which refers to Bapedal Regulation No. 01 In 1995, SNI 03-3985-2000, Employee Minister Regulation RI No. Per-04/Men/1980. Based on research that already done, founded the design for warehouse dimension of waste is 23 m × 22 m × 5 m with waste layout position appropriate with type of waste. The necessary of quantity for detector on this waste warehouse design are 56 each. The type of fire extinguisher that appropriate with this design is dry powder which containing natrium carbonate, alkali salts, with having each weight of 12 Kg about 18 units.

[Waste management in hospitals. Current situation in the state of North Rhine-Westphalia].

PubMed

Popp, W; Hansen, D; Hilgenhöner, M; Grandek, M; Heinemann, A; Blättler, T

2009-07-01

In 20 hospitals in North Rhine-Westphalia in-plant handling wastes and the delivery of the waste to the disposer were examined. Deficits were seen regarding risk assessment and operating instructions, support by company doctors, personal protection equipment, and break areas for the waste collecting personnel. Also the qualification of the waste management officer and his/her time contingent, correct declaration of the wastes, the training of the waste collecting personnel, the cleaning of multi-use containers and transportation vehicles, storage of the wastes at the collecting points, and the use of sharp collecting boxes were to be partly criticized. Consequences and recommendations are given, concerning the company's obligations (e.g., provide risk assessment, operating instructions), waste management officer (e.g., qualification, enough time contingent, regular inspections), waste collecting personnel (e.g., training courses), industrial safety (e.g., protection equipment, break area wash places), company doctors, transportation vehicles in the house (e.g., regular cleaning), one-way collectors (e.g., labelling at the site of the collection), multi-use collectors (e.g., cleaning), and compressing containers (e.g., larger maintenance openings).

The comparison of fossil carbon fraction and greenhouse gas emissions through an analysis of exhaust gases from urban solid waste incineration facilities.

PubMed

Kim, Seungjin; Kang, Seongmin; Lee, Jeongwoo; Lee, Seehyung; Kim, Ki-Hyun; Jeon, Eui-Chan

2016-10-01

In this study, in order to understand accurate calculation of greenhouse gas emissions of urban solid waste incineration facilities, which are major waste incineration facilities, and problems likely to occur at this time, emissions were calculated by classifying calculation methods into 3 types. For the comparison of calculation methods, the waste characteristics ratio, dry substance content by waste characteristics, carbon content in dry substance, and (12)C content were analyzed; and in particular, CO2 concentration in incineration gases and (12)C content were analyzed together. In this study, 3 types of calculation methods were made through the assay value, and by using each calculation method, emissions of urban solid waste incineration facilities were calculated then compared. As a result of comparison, with Calculation Method A, which used the default value as presented in the IPCC guidelines, greenhouse gas emissions were calculated for the urban solid waste incineration facilities A and B at 244.43 ton CO2/day and 322.09 ton CO2/day, respectively. Hence, it showed a lot of difference from Calculation Methods B and C, which used the assay value of this study. It is determined that this was because the default value as presented in IPCC, as the world average value, could not reflect the characteristics of urban solid waste incineration facilities. Calculation Method B indicated 163.31 ton CO2/day and 230.34 ton CO2/day respectively for the urban solid waste incineration facilities A and B; also, Calculation Method C indicated 151.79 ton CO2/day and 218.99 ton CO2/day, respectively. This study intends to compare greenhouse gas emissions calculated using (12)C content default value provided by the IPCC (Intergovernmental Panel on Climate Change) with greenhouse gas emissions calculated using (12)C content and waste assay value that can reflect the characteristics of the target urban solid waste incineration facilities. Also, the concentration and (12)C content were calculated by directly collecting incineration gases of the target urban solid waste incineration facilities, and greenhouse gas emissions of the target urban solid waste incineration facilities through this survey were compared with greenhouse gas emissions, which used the previously calculated assay value of solid waste.

Idea Bank.

ERIC Educational Resources Information Center

Talesnick, Irwin, Ed.

1984-01-01

Provides innovative ideas in biology, chemistry, and physics on the following topics: enzyme decomposition; chemical waste; time measurement; acid-base color magic; ball bouncing properties; heat; cell theory; and specimen boxes. Materials and procedures are listed when appropriate along with hints for expanding these ideas and investigations. (JM)

The rice F-box protein KISS ME DEADLY2 functions as a negative regulator of cytokinin signalling.

PubMed

Kim, Hyo Jung; Kieber, Joseph J; Schaller, G Eric

2013-01-01

Cytokinins are plant hormones that play critical roles in growth and development. We recently determined that the transcriptional response to cytokinin of Arabidopsis is modulated by the KISS ME DEADLY (KMD) family of F-box proteins. Here we demonstrate a conserved function for a member of the rice KMD family. Ectopic overexpression of OsKMD2 in Arabidopsis results in decreased cytokinin sensitivity based on a hypocotyl growth response assay, the decrease in sensitivity correlating with a decrease in the levels of the transcriptional regulator AtARR12. Furthermore, OsKMD2 directly interacts with AtARR12 based on yeast two-hybrid and co-immunoprecipitation assays. These results indicate that both monocots and dicots employ a similar KMD-dependent mechanism to regulate the transcriptional response to cytokinin.

Packaging, Transportation and Recycling of NPP Condenser Modules - 12262

DOE Office of Scientific and Technical Information (OSTI.GOV)

Polley, G.M.

2012-07-01

Perma-Fix was awarded contract from Energy Northwest for the packaging, transportation and disposition of the condenser modules, water boxes and miscellaneous metal, combustibles and water generated during the 2011 condenser replacement outage at the Columbia Generating Station. The work scope was to package the water boxes and condenser modules as they were removed from the facility and transfer them to the Perma-Fix Northwest facility for processing, recycle of metals and disposition. The condenser components were oversized and overweight (the condenser modules weighed ∼102,058 kg [225,000 lb]) which required special equipment for loading and transport. Additional debris waste was packaged inmore » inter-modals and IP-1 boxes for transport. A waste management plan was developed to minimize the generation of virtually any waste requiring landfill disposal. The Perma-Fix Northwest facility was modified to accommodate the ∼15 m [50-ft] long condenser modules and equipment was designed and manufactured to complete the disassembly, decontamination and release survey. The condenser modules are currently undergoing processing for free release to a local metal recycler. Over three millions pounds of metal will be recycled and over 95% of the waste generated during this outage will not require land disposal. There were several elements of this project that needed to be addressed during the preparation for this outage and the subsequent packaging, transportation and processing. - Staffing the project to support 24/7 generation of large components and other wastes. - The design and manufacture of the soft-sided shipping containers for the condenser modules that measured ∼15 m X 4 m X 3 m [50 ft X 13 ft X 10 ft] and weighed ∼102,058 kg [225,000 lbs] - Developing a methodology for loading the modules into the shipping containers. - Obtaining a transport vehicle for the modules. - Designing and modifying the processing facility. - Movement of the modules at the processing facility. If any of these issues were not adequately resolved prior to the start of the outage, costly delays would result and the re-start of the power plant could be impacted. The main focus of this project was to find successful methods for keeping this material out of the landfills and preserving the natural resources. In addition, this operation provided a significant cost savings to the public utility by minimizing landfill disposal. The onsite portion of the project has been completed without impact to the overall outage schedule. By the date of presentation, the majority of the waste from the condenser replacement project will have been processed and recycled. The goals for this project included helping Energy Northwest maintain the outage schedule, package and characterize waste compliantly, perform transportation activities in compliance with 49CFR (Ref-1), and minimize the waste disposal volume. During this condenser replacement project, over three millions pounds of waste was generated, packaged, characterized and transported without injury or incident. It is anticipated that 95% of the waste generated during this project will not require landfill disposal. All of the waste is scheduled to be processed, decontaminated and recycled by June of 2012. (authors)« less

PRODUCTION OF BIOBUTANOL FROM BIOMASS USING NOVEL MEMBRANE REACTOR

EPA Science Inventory

Food Waste Composition
Team REFUSE first performed sugar and starch assays on food waste from an on-campus cafeteria to determine its composition. The testing of the Chartwells food waste indicated the waste contained approximately 10% sugar and 25-30% starch co...

A DNA-binding protein from Candida albicans that binds to the RPG box of Saccharomyces cerevisiae and the telomeric repeat sequence of C. albicans.

PubMed

Ishii, N; Yamamoto, M; Lahm, H W; Iizumi, S; Yoshihara, F; Nakayama, H; Arisawa, M; Aoki, Y

1997-02-01

Electromobility shift assays with a DNA probe containing the Saccharomyces cerevisiae ENO1 RPG box identified a specific DNA-binding protein in total protein extracts of Candida albicans. The protein, named Rbf1p (RPG-box-binding protein 1), bound to other S. cerevisiae RPG boxes, although the nucleotide recognition profile was not completely the same as that of S. cerevisiae Rap 1p (repressor-activator protein 1), an RPG-box-binding protein. The repetitive sequence of the C. albicans chromosomal telomere also competed with RPG-box binding to Rbf1p. For further analysis, we purified Rbf1p 57,600-fold from C. albicans total protein extracts, raised mAbs against the purified protein and immunologically cloned the gene, whose ORF specified a protein of 527 aa. The bacterially expressed protein showed RPG-box-binding activity with the same profile as that of the purified one. The Rbf1p, containing two glutamine-rich regions that are found in many transcription factors, showed transcriptional activation capability in S. cerevisiae and was predominantly observed in nuclei. These results suggest that Rbf1p is a transcription factor with telomere-binding activity in C. albicans.

Characterization of Co(III) EDTA-Reducing Bacteria in Metal- and Radionuclide-Contaminated Groundwater

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gao, Weimin; Gentry, Terry J; Mehlhorn, Tonia L

The Waste Area Grouping 5 (WAG5) site at Oak Ridge National Laboratory has a potential to be a field site for evaluating the effectiveness of various bioremediation approaches and strategies. The site has been well studied in terms of its geological and geochemical properties over the past decade. However, despite the importance of microorganisms in bioremediation processes, the microbiological populations at the WAG5 site and their potential in bioremediation have not been similarly evaluated. In this study, we initiated research to characterize the microbial populations in WAG5 groundwater. Approximately 100 isolates from WAG5 groundwater were isolated and selected based onmore » colony morphology. Fifty-five unique isolates were identified by BOX-PCR and subjected to further characterization. 16S rRNA sequences indicated that these isolates belong to seventeen bacterial genera including Alcaligenes (1 isolate), Aquamonas (1), Aquaspirillum (1), Bacillus (10), Brevundimonas (5), Caulobacter (7), Dechloromonas (2), Janibacter (1), Janthinobacterium (2), Lactobacillus (1), Paenibacillus (4), Pseudomonas (9), Rhodoferax (1), Sphingomonas (1), Stenotrophomonas (6), Variovorax (2), and Zoogloea (1). Metal respiration assays identified several isolates, which phylogenically belong or are close to Caulobacter, Stenotrophomonas, Bacillus, Paenibacillus and Pseudomonas, capable of reducing Co(III)EDTA- to Co(II)EDTA{sup 2-} using the defined M1 medium under anaerobic conditions. In addition, using WAG5 groundwater directly as the inoculants, we found that organisms associated with WAG5 groundwater can reduce both Fe(III) and Co(III) under anaerobic conditions. Further assays were then performed to determine the optimal conditions for Co(III) reduction. These assays indicated that addition of various electron donors including ethanol, lactate, methanol, pyruvate, and acetate resulted in metal reduction. These experiments will provide useful background information for future bioremediation field experiments at the WAG5 site.« less

Remaining Sites Verification Package for the 100-F-44:2, Discovery Pipeline Near 108-F Building, Waste Site Reclassification Form 2007-006

DOE Office of Scientific and Technical Information (OSTI.GOV)

J. M. Capron

2008-05-30

The 100-F-44:2 waste site is a steel pipeline that was discovered in a junction box during confirmatory sampling of the 100-F-26:4 pipeline from December 2004 through January 2005. The 100-F-44:2 pipeline feeds into the 100-F-26:4 subsite vitrified clay pipe (VCP) process sewer pipeline from the 108-F Biology Laboratory at the junction box. In accordance with this evaluation, the confirmatory sampling results support a reclassification of this site to No Action. The current site conditions achieve the remedial action objectives and the corresponding remedial action goals established in the Remaining Sites ROD. The results of confirmatory sampling show that residual contaminantmore » concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also demonstrate that residual contaminant concentrations are protective of groundwater and the Columbia River.« less

High level waste tank closure project: ALARA applications at the Idaho National Engineering and Environmental Laboratory.

PubMed

Aitken, Steven B; Butler, Richard; Butterworth, Steven W; Quigley, Keith D

2005-05-01

Bechtel BWXT Idaho, Maintenance and Operating Contractor for the Department of Energy at the Idaho National Engineering and Environmental Laboratory, has emptied, cleaned, and sampled six of the eleven 1.135 x 10(6) L high level waste underground storage tanks at the Idaho Nuclear Technology and Engineering Center, well ahead of the State of Idaho Consent Order cleaning schedule. Cleaning of a seventh tank is expected to be complete by the end of calendar year 2004. The tanks, with associated vaults, valve boxes, and distribution systems, are being closed to meet Resource Conservation and Recovery Act regulations and Department of Energy orders. The use of remotely operated equipment placed in the tanks through existing tank riser access points, sampling methods and application of as-low-as-reasonably-achievable (ALARA) principles have proven effective in keeping personnel dose low during equipment removal, tank, vault, and valve box cleaning, and sampling activities, currently at 0.03 Sv.

A semi-automated luminescence based standard membrane feeding assay identifies novel small molecules that inhibit transmission of malaria parasites by mosquitoes

PubMed Central

Vos, Martijn W.; Stone, Will J. R.; Koolen, Karin M.; van Gemert, Geert-Jan; van Schaijk, Ben; Leroy, Didier; Sauerwein, Robert W.; Bousema, Teun; Dechering, Koen J.

2015-01-01

Current first-line treatments for uncomplicated falciparum malaria rapidly clear the asexual stages of the parasite, but do not fully prevent parasite transmission by mosquitoes. The standard membrane feeding assay (SMFA) is the biological gold standard assessment of transmission reducing activity (TRA), but its throughput is limited by the need to determine mosquito infection status by dissection and microscopy. Here we present a novel dissection-free luminescence based SMFA format using a transgenic Plasmodium falciparum reporter parasite without resistance to known antimalarials and therefore unrestricted in its utility in compound screening. Analyses of sixty-five compounds from the Medicines for Malaria Venture validation and malaria boxes identified 37 compounds with high levels of TRA (>80%); different assay modes allowed discrimination between gametocytocidal and downstream modes of action. Comparison of SMFA data to published assay formats for predicting parasite infectivity indicated that individual in vitro screens show substantial numbers of false negatives. These results highlight the importance of the SMFA in the screening pipeline for transmission reducing compounds and present a rapid and objective method. In addition we present sixteen diverse chemical scaffolds from the malaria box that may serve as a starting point for further discovery and development of malaria transmission blocking drugs. PMID:26687564

Box-Behnken design approach towards optimization of activated carbon synthesized by co-pyrolysis of waste polyester textiles and MgCl2

NASA Astrophysics Data System (ADS)

Yuan, Zhihang; Xu, Zhihua; Zhang, Daofang; Chen, Weifang; Zhang, Tianqi; Huang, Yuanxing; Gu, Lin; Deng, Haixuan; Tian, Danqi

2018-01-01

Pyrolysis activation of waste polyester textiles (WPT) was regarded as a sustainable technique to synthesize multi-pore activated carbons. MgO-template method of using MgCl2 as the template precursor was employed, which possessed the advantages of ideal pore-forming effect and efficient preparation process. The response surface methodology coupled with Box-Behnken design (BBD) was conducted to study the interaction between different variables and optimized preparation conditions of waste polyester textiles based activated carbons. Derived from BBD design results, carbonization temperature was the most significant individual factor. And the maximum specific surface area of 1364 m2/g, which presented a good agreement with the predicted response values(1315 m2/g), was obtained at mixing ratio in MgCl2/WPT, carbonization temperature and time of 5:1, 900 °C and 90 min, respectively. Furthermore, the physicochemical properties of the sample prepared under optimal conditions were carried on utilizing nitrogen adsorption/desorption isotherms, EA, XRD, SEM and FTIR. In addition, the pore-forming mechanism was mainly attributed to the tendency of carbon layer coating on MgO to form pore walls after elimination of MgO and the strong dehydration effect of MgCl2 on WPT.

Statistical Evaluation and Optimization of Factors Affecting the Leaching Performance of Copper Flotation Waste

PubMed Central

Çoruh, Semra; Elevli, Sermin; Geyikçi, Feza

2012-01-01

Copper flotation waste is an industrial by-product material produced from the process of manufacturing copper. The main concern with respect to landfilling of copper flotation waste is the release of elements (e.g., salts and heavy metals) when in contact with water, that is, leaching. Copper flotation waste generally contains a significant amount of Cu together with trace elements of other toxic metals, such as Zn, Co, and Pb. The release of heavy metals into the environment has resulted in a number of environmental problems. The aim of this study is to investigate the leaching characteristics of copper flotation waste by use of the Box-Behnken experimental design approach. In order to obtain the optimized condition of leachability, a second-order model was examined. The best leaching conditions achieved were as follows: pH = 9, stirring time = 5 min, and temperature = 41.5°C. PMID:22629194

Statistical evaluation and optimization of factors affecting the leaching performance of copper flotation waste.

PubMed

Coruh, Semra; Elevli, Sermin; Geyikçi, Feza

2012-01-01

Copper flotation waste is an industrial by-product material produced from the process of manufacturing copper. The main concern with respect to landfilling of copper flotation waste is the release of elements (e.g., salts and heavy metals) when in contact with water, that is, leaching. Copper flotation waste generally contains a significant amount of Cu together with trace elements of other toxic metals, such as Zn, Co, and Pb. The release of heavy metals into the environment has resulted in a number of environmental problems. The aim of this study is to investigate the leaching characteristics of copper flotation waste by use of the Box-Behnken experimental design approach. In order to obtain the optimized condition of leachability, a second-order model was examined. The best leaching conditions achieved were as follows: pH = 9, stirring time = 5 min, and temperature = 41.5 °C.

40 CFR 246.200-6 - Recommended procedures: Storage.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Recommended procedures: Storage. 246....200-6 Recommended procedures: Storage. Among the alternatives for paper storage are on-site bailing, the use of stationary compactors, or storage in corrugated boxes or normal waste containers. Stored...

40 CFR 246.200-6 - Recommended procedures: Storage.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Recommended procedures: Storage. 246....200-6 Recommended procedures: Storage. Among the alternatives for paper storage are on-site bailing, the use of stationary compactors, or storage in corrugated boxes or normal waste containers. Stored...

40 CFR 246.200-6 - Recommended procedures: Storage.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 25 2014-07-01 2014-07-01 false Recommended procedures: Storage. 246....200-6 Recommended procedures: Storage. Among the alternatives for paper storage are on-site bailing, the use of stationary compactors, or storage in corrugated boxes or normal waste containers. Stored...

40 CFR 246.200-6 - Recommended procedures: Storage.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 26 2012-07-01 2011-07-01 true Recommended procedures: Storage. 246....200-6 Recommended procedures: Storage. Among the alternatives for paper storage are on-site bailing, the use of stationary compactors, or storage in corrugated boxes or normal waste containers. Stored...

40 CFR 246.200-6 - Recommended procedures: Storage.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Recommended procedures: Storage. 246....200-6 Recommended procedures: Storage. Among the alternatives for paper storage are on-site bailing, the use of stationary compactors, or storage in corrugated boxes or normal waste containers. Stored...

Analysis of a cis-Acting Element Involved in Regulation by Estrogen of Human Angiotensinogen Gene Expression.

PubMed

Zhao, Yan-Yan; Sun, Kai-Lai; Ashok, Kumar

1998-01-01

The work was aimed to identify the estrogen responsive element in the human angiotensinogen gene. The nucleotide sequence between the transcription initiation site and TATA box in angiotensinogen gene promoter was found to be strongly homologous with the consensus estrogen responsive element. This sequence was confirmed as the estrogen responsive element (HAG ERE) by electrophoretic mobility shift assay. The recombinant expression vectors were constructed in which chloramphenicol acetyltransferase (CAT) reporter gene was driven by angiotensinogen core promoter with HAG ERE of by TK core promoter with multiplied HAG ERE, and were used in cotransfection with the human estrogen receptor expression vector into HepG(2) cells; CAT assays showed an increase of the CAT activity on 17beta-estradiol treatment in those transfectants. These results suggest that the human angiotensinogen gene is transcriptionally up-regulated by estrogen through the estrogen responsive element near TATA box of the promoter.

Retraction: Myostatin Induces Degradation of Sarcomeric Proteins through a Smad3 Signaling Mechanism During Skeletal Muscle Wasting

PubMed Central

Lokireddy, Sudarsanareddy; McFarlane, Craig; Ge, Xiaojia; Zhang, Huoming; Sze, Siu Kwan; Sharma, Mridula

2011-01-01

Ubiquitination-mediated proteolysis is a hallmark of skeletal muscle wasting manifested in response to negative growth factors, including myostatin. Thus, the characterization of signaling mechanisms that induce the ubiquitination of intracellular and sarcomeric proteins during skeletal muscle wasting is of great importance. We have recently characterized myostatin as a potent negative regulator of myogenesis and further demonstrated that elevated levels of myostatin in circulation results in the up-regulation of the muscle-specific E3 ligases, Atrogin-1 and muscle ring finger protein 1 (MuRF1). However, the exact signaling mechanisms by which myostatin regulates the expression of Atrogin-1 and MuRF1, as well as the proteins targeted for degradation in response to excess myostatin, remain to be elucidated. In this report, we have demonstrated that myostatin signals through Smad3 (mothers against decapentaplegic homolog 3) to activate forkhead box O1 and Atrogin-1 expression, which further promotes the ubiquitination and subsequent proteasome-mediated degradation of critical sarcomeric proteins. Smad3 signaling was dispensable for myostatin-dependent overexpression of MuRF1. Although down-regulation of Atrogin-1 expression rescued approximately 80% of sarcomeric protein loss induced by myostatin, only about 20% rescue was seen when MuRF1 was silenced, implicating that Atrogin-1 is the predominant E3 ligase through which myostatin manifests skeletal muscle wasting. Furthermore, we have highlighted that Atrogin-1 not only associates with myosin heavy and light chain, but it also ubiquitinates these sarcomeric proteins. Based on presented data we propose a model whereby myostatin induces skeletal muscle wasting through targeting sarcomeric proteins via Smad3-mediated up-regulation of Atrogin-1 and forkhead box O1. PMID:21964591

Mutations of the phage lambda nutL region that prevent the action of Nun, a site-specific transcription termination factor.

PubMed Central

Baron, J; Weisberg, R A

1992-01-01

Phage HK022 encodes a protein, Nun, that promotes transcription termination within the pL and pR operons of its relative, phage lambda. The lambda sequences required for termination had previously been shown to overlap the nut sites, which are essential for transcription antitermination during normal lambda growth. To further specify the Nun target and to determine its relation to the nut sites, we constructed deletion and base substitution mutations of the lambda nutL region and measured Nun-dependent reduction of the expression of a downstream reporter gene. The shortest construct that retained full Nun responsiveness was a 42-bp segment that included both boxA and boxB, sequences that have been implicated in lambda antitermination. Deletion of boxA reduced Nun termination, and deletion of both sequences eliminated Nun termination. Base substitutions in boxA and the proximal portion of boxB impaired Nun termination, while base substitutions between boxA and boxB, in the distal portion of boxB, and immediately downstream from boxB had no appreciable effect. The termination defect of all of the base substitution mutations was relieved by increasing the level of Nun protein; in contrast, the deletions and a multiple-base substitution did not regain full Nun responsiveness at elevated Nun concentrations. We also asked if these mutant nut regions retained their ability to interact with N, the lambda-encoded antitermination protein. A qualitative assay showed that mutations within boxA or boxB reduced interaction, while mutations outside boxA and boxB did not. These data show that (i) the recognition sites for N and Nun overlap to a very considerable extent but are probably not identical and (ii) a high concentration of Nun promotes its interaction with mutant nut sites, a behavior also reported to be characteristic of N. PMID:1532174

Has the question of e-waste opened a Pandora's box? An overview of unpredictable issues and challenges.

PubMed

Bakhiyi, Bouchra; Gravel, Sabrina; Ceballos, Diana; Flynn, Michael A; Zayed, Joseph

2018-01-01

Despite regulatory efforts and position papers, electrical and electronic waste (e-waste) remains ill-managed as evidenced by the extremely low rates of proper e-waste recycling (e-recycling) worldwide, ongoing illegal shipments to developing countries and constantly reported human health issues and environmental pollution. The objectives of this review are, first, to expose the complexity of e-waste problems, and then to suggest possible upstream and downstream solutions. Exploring e-waste issues is akin to opening a Pandora's box. Thus, a review of prevailing e-waste management practices reveals complex and often intertwined gaps, issues and challenges. These include the absence of any consistent definition of e-waste to date, a prevalent toxic potential still involving already banned or restricted hazardous components such as heavy metals and persistent and bioaccumulative organic compounds, a relentless growth in e-waste volume fueled by planned obsolescence and unsustainable consumption, problematic e-recycling processes, a fragile formal e-recycling sector, sustained and more harmful informal e-recycling practices, and more convoluted and unpredictable patterns of illegal e-waste trade. A close examination of the e-waste legacy contamination reveals critical human health concerns, including significant occupational exposure during both formal and informal e-recycling, and persistent environmental contamination, particularly in some developing countries. However, newly detected e-waste contaminants as well as unexpected sources and environmental fates of contaminants are among the emerging issues that raise concerns. Moreover, scientific knowledge gaps remain regarding the complexity and magnitude of the e-waste legacy contamination, specifically, a comprehensive characterization of e-waste contaminants, information on the scale of legacy contamination in developing countries and on the potential environmental damage in developed countries, and a stronger body of evidence of adverse health effects specifically ascribed to e-waste contaminants. However, the knowledge accumulated to date is sufficient to raise awareness and concern among all stakeholders. Potential solutions to curb e-waste issues should be addressed comprehensively, by focusing on two fronts: upstream and downstream. Potential upstream solutions should focus on more rational and eco-oriented consumer habits in order to decrease e-waste quantities while fostering ethical and sustained commitments from manufacturers, which include a limited usage of hazardous compounds and an optimal increase in e-waste recyclability. At the downstream level, solutions should include suitable and pragmatic actions to progressively reduce the illegal e-waste trade particularly through international cooperation and coordination, better enforcement of domestic laws, and monitoring in both exporting and receiving countries, along with the supervised integration of the informal sector into the recycling system of developing countries and global expansion of formal e-waste collection and recycling activities. Downstream solutions should also introduce stronger reverse logistics, together with upgraded, more affordable, and eco-friendly and worker-friendly e-recycling technologies to ensure that benefits are derived fully and safely from the great economic potential of e-waste. Copyright © 2017 Elsevier Ltd. All rights reserved.

Environmental Technology Verification Report for Private Pallet Security Systems, LLC. MultiTrack™ Layered Tracking Systems

EPA Science Inventory

This test simulated shipments of hazardous waste contained in polyethylene (poly) drums, metal drums, and corrugated boxes through routine land transportation routes and across international ports of entry in the El Paso/Ciudad Juarez trade area. RFID tags were attached to four ...

40 CFR 62.14104 - Requirements for municipal waste combustor operating practices.

Code of Federal Regulations, 2013 CFR

2013-07-01

...), proceed in accordance with ASME PTC 4.1-1964 (Reaffirmed 1991), Power Test Codes: Test Code for Steam Generating Units (with 1968 and 1969 Addenda). For design, construction, installation, calibration, and use... Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You may...

40 CFR 62.14104 - Requirements for municipal waste combustor operating practices.

Code of Federal Regulations, 2014 CFR

2014-07-01

...), proceed in accordance with ASME PTC 4.1-1964 (Reaffirmed 1991), Power Test Codes: Test Code for Steam Generating Units (with 1968 and 1969 Addenda). For design, construction, installation, calibration, and use... Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You may...

40 CFR 62.14104 - Requirements for municipal waste combustor operating practices.

Code of Federal Regulations, 2010 CFR

2010-07-01

...), proceed in accordance with ASME PTC 4.1-1964 (Reaffirmed 1991), Power Test Codes: Test Code for Steam Generating Units (with 1968 and 1969 Addenda). For design, construction, installation, calibration, and use... Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You may...

40 CFR 62.14104 - Requirements for municipal waste combustor operating practices.

Code of Federal Regulations, 2012 CFR

2012-07-01

...), proceed in accordance with ASME PTC 4.1-1964 (Reaffirmed 1991), Power Test Codes: Test Code for Steam Generating Units (with 1968 and 1969 Addenda). For design, construction, installation, calibration, and use... Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You may...

40 CFR 62.14104 - Requirements for municipal waste combustor operating practices.

Code of Federal Regulations, 2011 CFR

2011-07-01

...), proceed in accordance with ASME PTC 4.1-1964 (Reaffirmed 1991), Power Test Codes: Test Code for Steam Generating Units (with 1968 and 1969 Addenda). For design, construction, installation, calibration, and use... Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You may...

CITY IN A BOX: A NEW PARADIGM FOR SUSTAINABLE LIVING

EPA Science Inventory

Global carbon emissions generated by buildings are estimated at between 40 to 50 percent, 25 percent from transportation and about 25 percent from industrial sources. In addition, cities are estimated to be responsible for as much as 70 percent of the waste impacting the envir...

36. MAIN CANAL Photographic copy of construction drawing dated ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

36. MAIN CANAL - Photographic copy of construction drawing dated 1907 (from Record Group 115, Box 17, Denver Branch of the National Archives, Denver) WASTE GATES ABOVE DARK CANYON SIPHON - Carlsbad Irrigation District, Main Canal, 4 miles North to 12 miles Southeast of Carlsbad, Carlsbad, Eddy County, NM

A novel activity of HMG domains: promotion of the triple-stranded complex formation between DNA containing (GGA/TCC)11 and d(GGA)11 oligonucleotides.

PubMed Central

Suda, T; Mishima, Y; Takayanagi, K; Asakura, H; Odani, S; Kominami, R

1996-01-01

The high mobility group protein (HMG)-box is a DNA-binding domain found in many proteins that bind preferentially to DNA of irregular structures in a sequence-independent manner and can bend the DNA. We show here that GST-fusion proteins of HMG domains from HMG1 and HMG2 promote a triple-stranded complex formation between DNA containing the (GGA/TCC)11 repeat and oligonucleotides of d(GGA)11 probably due to G:G base pairing. The activity is to reduce association time and requirements of Mg2+ and oligonucleotide concentrations. The HMG box of SRY, the protein determining male-sex differentiation, also has the activity, suggesting that it is not restricted to the HMG-box domains derived from HMG1/2 but is common to those from other members of the HMG-box family of proteins. Interestingly, the box-AB and box-B of HMG1 bend DNA containing the repeat, but SRY fails to bend in a circularization assay. The difference suggests that the two activities of association-promotion and DNA bending are distinct. These results suggest that the HMG-box domain has a novel activity of promoting the association between GGA repeats which might be involved in higher-order architecture of chromatin. PMID:8972860

Determination of estrogenic potential in waste water without sample extraction.

PubMed

Avberšek, Miha; Žegura, Bojana; Filipič, Metka; Uranjek-Ževart, Nataša; Heath, Ester

2013-09-15

This study describes the modification of the ER-Calux assay for testing water samples without sample extraction (NE-(ER-Calux) assay). The results are compared to those obtained with ER-Calux assay and a theoretical estrogenic potential obtained by GC-MSD. For spiked tap and waste water samples there was no statistical difference between estrogenic potentials obtained by the three methods. Application of NE-(ER-Calux) to "real" influent and effluents from municipal waste water treatment plants and receiving surface waters found that the NE-(ER-Calux) assay gave higher values compared to ER-Calux assay and GC-MSD. This is explained by the presence of water soluble endocrine agonists that are usually removed during extraction. Intraday dynamics of the estrogenic potential of a WWTP influent and effluent revealed an increase in the estrogenic potential of the influent from 12.9 ng(EEQ)/L in the morning to a peak value of 40.0 ng(EEQ)/L in the afternoon. The estrogenic potential of the effluent was

Dismantling of the PETRA glove box: tritium contamination and inventory assessment

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wagner, R.

2015-03-15

The PETRA facility is the first installation in which experiments with tritium were carried out at the Tritium Laboratory Karlsruhe. After completion of two main experimental programs, the decommissioning of PETRA was initiated with the aim to reuse the glove box and its main still valuable components. A decommissioning plan was engaged to: -) identify the source of tritium release in the glove box, -) clarify the status of the main components, -) assess residual tritium inventories, and -) de-tritiate the components to be disposed of as waste. Several analytical techniques - calorimetry on small solid samples, wipe test followedmore » by liquid scintillation counting for surface contamination assessment, gas chromatography on gaseous samples - were deployed and cross-checked to assess the remaining tritium inventories and initiate the decommissioning process. The methodology and the main outcomes of the numerous different tritium measurements are presented and discussed. (authors)« less

Remaining Sites Verification Package for the 331 Life Sciences Laboratory Drain Field Septic System, Waste Site Reclassification Form 2008-020

DOE Office of Scientific and Technical Information (OSTI.GOV)

J. M. Capron

2008-10-16

The 331 Life Sciences Laboratory Drain Field (LSLDF) septic system waste site consists of a diversion chamber, two septic tanks, a distribution box, and a drain field. This septic system was designed to receive sanitary waste water, from animal studies conducted in the 331-A and 331-B Buildings, for discharge into the soil column. However, field observations and testing suggest the 331 LSLDF septic system did not receive any discharges. In accordance with this evaluation, the confirmatory sampling results support a reclassification of the 331 LSLDF waste site to No Action. This site does not have a deep zone or othermore » condition that would warrant an institutional control in accordance with the 300-FF-2 ROD under the industrial land use scenario.« less

Characterization of cis-acting elements required for autorepression of the equine herpesvirus 1 IE gene

PubMed Central

Kim, Seongman; Dai, Gan; O’Callaghan, Dennis J.; Kim, Seong Kee

2012-01-01

The immediate-early protein (IEP), the major regulatory protein encoded by the IE gene of equine herpesvirus 1 (EHV-1), plays a crucial role as both transcription activator and repressor during a productive lytic infection. To investigate the mechanism by which the EHV-1 IEP inhibits its own promoter, IE promoter-luciferase reporter plasmids containing wild-type and mutant IEP-binding site (IEBS) were constructed and used for luciferase reporter assays. The IEP inhibited transcription from its own promoter in the presence of a consensus IEBS (5’-ATCGT-3’) located near the transcription initiation site but did not inhibit when the consensus sequence was deleted. To determine whether the distance between the TATA box and the IEBS affects transcriptional repression, the IEBS was displaced from the original site by the insertion of synthetic DNA sequences. Luciferase reporter assays revealed that the IEP is able to repress its own promoter when the IEBS is located within 26-bp from the TATA box. We also found that the proper orientation and position of the IEBS were required for the repression by the IEP. Interestingly, the level of repression was significantly reduced when a consensus TATA sequence was deleted from the promoter region, indicating that the IEP efficiently inhibits its own promoter in a TATA box-dependent manner. Taken together, these results suggest that the EHV-1 IEP delicately modulates autoregulation of its gene through the consensus IEBS that is near the transcription initiation site and the TATA box. PMID:22265772

Characterization of cis-acting elements required for autorepression of the equine herpesvirus 1 IE gene.

PubMed

Kim, Seongman; Dai, Gan; O'Callaghan, Dennis J; Kim, Seong Kee

2012-04-01

The immediate-early protein (IEP), the major regulatory protein encoded by the IE gene of equine herpesvirus 1 (EHV-1), plays a crucial role as both transcription activator and repressor during a productive lytic infection. To investigate the mechanism by which the EHV-1 IEP inhibits its own promoter, IE promoter-luciferase reporter plasmids containing wild-type and mutant IEP-binding site (IEBS) were constructed and used for luciferase reporter assays. The IEP inhibited transcription from its own promoter in the presence of a consensus IEBS (5'-ATCGT-3') located near the transcription initiation site but did not inhibit when the consensus sequence was deleted. To determine whether the distance between the TATA box and the IEBS affects transcriptional repression, the IEBS was displaced from the original site by the insertion of synthetic DNA sequences. Luciferase reporter assays revealed that the IEP is able to repress its own promoter when the IEBS is located within 26-bp from the TATA box. We also found that the proper orientation and position of the IEBS were required for the repression by the IEP. Interestingly, the level of repression was significantly reduced when a consensus TATA sequence was deleted from the promoter region, indicating that the IEP efficiently inhibits its own promoter in a TATA box-dependent manner. Taken together, these results suggest that the EHV-1 IEP delicately modulates autoregulation of its gene through the consensus IEBS that is near the transcription initiation site and the TATA box. Copyright © 2012. Published by Elsevier B.V.

Development of an integrated transuranic waste management system for a large research facility: NUCEF

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mineo, Hideaki; Matsumura, Tatsuro; Takeshita, Isao

1997-03-01

The Nuclear Fuel Cycle Safety Engineering Research Facility (NUCEF) is a large complex of research facilities where transuranic (TRU) elements are used. Liquid and solid waste containing TRU elements is generated mainly in the treatment of fuel for critical experiments and in the research of reprocessing and TRU waste management in hot cells and glove boxes. The rational management of TRU wastes is a very important issue not only for NUCEF but also for Japan. An integrated TRU waste management system is being developed with NUCEF as the test bed. The basic policy for establishing the system is to classifymore » wastes by TRU concentration, to reduce waste volume, and to maximize reuse of TRU elements. The principal approach of the development program is to apply the outcomes of the research carried out in NUCEF. Key technologies are TRU measurement for classification of solid wastes and TRU separation and volume reduction for organic and aqueous wastes. Some technologies required for treating the wastes specific to the research activities in NUCEF need further development. Specifically, the separation and stabilization technologies for americium recovery from concentrated aqueous waste, which is generated in dissolution of mixed oxide when preparing fuel for critical experiments, needs further research.« less

Effect of leachate recirculation on mesophilic anaerobic digestion of food waste.

PubMed

Shahriari, Haleh; Warith, Mostafa; Hamoda, Mohamed; Kennedy, Kevin J

2012-03-01

The effects of using untreated leachate for supplemental water addition and liquid recirculation on anaerobic digestion of food waste was evaluated by combining cyclic water recycle operations with batch mesophilic biochemical methane potential (BMP) assays. Cyclic BMP assays indicated that using an appropriate fraction of recycled leachate and fresh make up water can stimulate methanogenic activity and enhance biogas production. Conversely increasing the percentage of recycled leachate in the make up water eventually causes methanogenic inhibition and decrease in the rate of food waste stabilization. The decrease in activity is exacerbated as the number cycles increases. Inhibition is possibly attributed to accumulation and elevated concentrations of ammonia as well as other waste by products in the recycled leachate that inhibit methanogenesis. Copyright © 2011 Elsevier Ltd. All rights reserved.

SLFL Genes Participate in the Ubiquitination and Degradation Reaction of S-RNase in Self-compatible Peach.

PubMed

Chen, Qiuju; Meng, Dong; Gu, Zhaoyu; Li, Wei; Yuan, Hui; Duan, Xuwei; Yang, Qing; Li, Yang; Li, Tianzhong

2018-01-01

It has been proved that the gametophytic self-incompatibility (GSI), mainly exists in Rosaceae and Solanaceae, is controlled by S genes, which are two tightly linked genes located at highly polymorphic S -locus: the S-RNase for pistil specificity and the F-box gene ( SFB/SLF ) for pollen specificity, respectively. However, the roles of those genes in SI of peach are still a subject of extensive debate. In our study, we selected 37 representative varieties according to the evolution route of peach and identified their S genotypes. We cloned pollen determinant genes mutated PperSFB1m, PperSFB2m, PperSFB4m , and normal PperSFB2 , and style determinant genes PperS1-RNase, PperS2-RNase, PperS2m-RNase , and PperS4-RNase . The mutated PperSFBs encode truncated SFB proteins due to a fragment insertion. The truncated PperSFBs and normal PperSFB2 interacted with PperS-RNases demonstrated by Y2H. Normal PperSFB2 was divided into four parts: box, box-V1, V1-V2, and HVa-HVb. The box domain of PperSFB2 did not interact with PperS-RNases, both of the box-V1 and V1-V2 had interactions with PperS-RNases, while the hypervariable region of PperSFB2 HVa-HVb only interacted with PperS2-RNase showed by Y2H and BiFC assay. Bioinformatics analysis of peach genome revealed that there were other F-box genes located at S-locus, and of which three F-box genes were specifically expressed in pollen, named as PperSLFL1, PperSLFL2 , and PperSLFL3 , respectively. In phylogenetic analysis PperSLFLs clustered with Maloideae SFBB genes, and PperSFB genes were clustered into the other group with other SFB genes of Prunus . Protein interaction analysis revealed that the three PperSLFLs interacted with PperSSK1 and PperS-RNases with no allelic specificity. In vitro ubiquitination assay showed that PperSLFLs could tag ubiquitin molecules onto PperS-RNases. The above results suggest that three PperSLFLs are the appropriate candidates for the "general inhibitor," which would inactivate the S-RNases in pollen tubes, involved in the self-incompatibility of peach.

SLFL Genes Participate in the Ubiquitination and Degradation Reaction of S-RNase in Self-compatible Peach

PubMed Central

Chen, Qiuju; Meng, Dong; Gu, Zhaoyu; Li, Wei; Yuan, Hui; Duan, Xuwei; Yang, Qing; Li, Yang; Li, Tianzhong

2018-01-01

It has been proved that the gametophytic self-incompatibility (GSI), mainly exists in Rosaceae and Solanaceae, is controlled by S genes, which are two tightly linked genes located at highly polymorphic S-locus: the S-RNase for pistil specificity and the F-box gene (SFB/SLF) for pollen specificity, respectively. However, the roles of those genes in SI of peach are still a subject of extensive debate. In our study, we selected 37 representative varieties according to the evolution route of peach and identified their S genotypes. We cloned pollen determinant genes mutated PperSFB1m, PperSFB2m, PperSFB4m, and normal PperSFB2, and style determinant genes PperS1-RNase, PperS2-RNase, PperS2m-RNase, and PperS4-RNase. The mutated PperSFBs encode truncated SFB proteins due to a fragment insertion. The truncated PperSFBs and normal PperSFB2 interacted with PperS-RNases demonstrated by Y2H. Normal PperSFB2 was divided into four parts: box, box-V1, V1-V2, and HVa-HVb. The box domain of PperSFB2 did not interact with PperS-RNases, both of the box-V1 and V1-V2 had interactions with PperS-RNases, while the hypervariable region of PperSFB2 HVa-HVb only interacted with PperS2-RNase showed by Y2H and BiFC assay. Bioinformatics analysis of peach genome revealed that there were other F-box genes located at S-locus, and of which three F-box genes were specifically expressed in pollen, named as PperSLFL1, PperSLFL2, and PperSLFL3, respectively. In phylogenetic analysis PperSLFLs clustered with Maloideae SFBB genes, and PperSFB genes were clustered into the other group with other SFB genes of Prunus. Protein interaction analysis revealed that the three PperSLFLs interacted with PperSSK1 and PperS-RNases with no allelic specificity. In vitro ubiquitination assay showed that PperSLFLs could tag ubiquitin molecules onto PperS-RNases. The above results suggest that three PperSLFLs are the appropriate candidates for the “general inhibitor,” which would inactivate the S-RNases in pollen tubes, involved in the self-incompatibility of peach. PMID:29520292

Geotechnical, geological, and selected radionuclide retention characteristics of the radioactive waste disposal site near the Farallon Islands

USGS Publications Warehouse

Booth, J.S.; Winters, W.J.; Poppe, L.J.; Neiheisel, J.; Dyer, R.S.

1989-01-01

A geotechnical and geological investigation of the Farallon Islands low-level radioactive waste (LLW) disposal area was conducted to qualitatively assess the host sediments' relative effectiveness as a barrier to radionuclide migration, to estimate the portion of the barrier that is in contact with the waste packages at the three primary disposal sites, and to provide a basic physical description of the sediments. Box cores recovered from within the general disposal area at depths of 500, 1000, and 1500 m were subcored to provide samples (~30 cm in length) for detailed descriptions, textural and mineralogical analyses, and a suite of geotechnical tests (index property, CRS consolidation, and CIU triaxial compression). -from Authors

29 CFR 776.20 - “Goods.”

Code of Federal Regulations, 2013 CFR

2013-07-01

..., bills of lading, checks, drafts, negotiable notes and other commercial paper. 26 “Goods” includes gold... or rented; 33 ice; 34 containers, as, for example, cigar boxes or wrapping paper and packing... of the four dissenting justices in 10 E. 40th St. Bldg. v. Callus, 325 U.S. at p. 586. Waste paper...

29 CFR 776.20 - “Goods.”

Code of Federal Regulations, 2014 CFR

2014-07-01

..., bills of lading, checks, drafts, negotiable notes and other commercial paper. 26 “Goods” includes gold... or rented; 33 ice; 34 containers, as, for example, cigar boxes or wrapping paper and packing... of the four dissenting justices in 10 E. 40th St. Bldg. v. Callus, 325 U.S. at p. 586. Waste paper...

29 CFR 776.20 - “Goods.”

Code of Federal Regulations, 2012 CFR

2012-07-01

..., bills of lading, checks, drafts, negotiable notes and other commercial paper. 26 “Goods” includes gold... or rented; 33 ice; 34 containers, as, for example, cigar boxes or wrapping paper and packing... of the four dissenting justices in 10 E. 40th St. Bldg. v. Callus, 325 U.S. at p. 586. Waste paper...

Recovering lead from cupel waste generated in gold analysis by Pb-Fire assay.

PubMed

Cerceau, Cristiane Isaac; Carvalho, Cornélio de Freitas; Rabelo, Ana Carolina Silveira; Dos Santos, Cláudio Gouvea; Gonçalves, Sabrina Mayra Dias; Varejão, Eduardo Vinícius Vieira

2016-12-01

Because of its precision and accuracy, Pb-Fire assay is the most employed method for gold analysis in geological materials. At the second stage of the method, namely cupellation, lead is oxidized to PbO which is absorbed by the cupel, leading to metallic gold as a tiny bend at the bottom of the recipient. After cupellation, cupel becomes highly contaminated with lead, making its disposal a serious risk of environmental contamination. In the present work, a leaching process for removing lead from cupel waste is proposed, which allowed for removing 96% of PbO by weight. After a precipitation step, 92.0% of lead was recovered from leachates in the form of PbSO 4 . Lead in the solid wastes left by the extraction was above the limit established by Brazilian legislation and these were classified as non-hazardous. Finally, secondary effluents generated after the precipitation step presented lead content more than twenty times lower than that of leachates from cupel waste. Tons of cupel waste are annually generated from gold analysis by Pb-Fire assay. Thus, the proposed method can contribute to prevent the discharge of high amounts of lead into the environment. Also, recovery of lead can help to partially meet the industrial demand for lead compounds. Copyright © 2016 Elsevier Ltd. All rights reserved.

Recognition of a wide-range of S-RNases by S locus F-box like 2, a general-inhibitor candidate in the Prunus-specific S-RNase-based self-incompatibility system.

PubMed

Matsumoto, Daiki; Tao, Ryutaro

2016-07-01

Many species in the Rosaceae, the Solanaceae, and the Plantaginaceae exhibit S-RNase-based gametophytic self-incompatibility (GSI). This system comprises S-ribonucleases (S-RNases) as the pistil S determinant and a single or multiple F-box proteins as the pollen S determinants. In Prunus, pollen specificity is determined by a single S haplotype-specific F-box protein (SFB). The results of several studies suggested that SFB exerts cognate S-RNase cytotoxicity, and a hypothetical general inhibitor (GI) is assumed to detoxify S-RNases in non-specific manner unless it is affected by SFB. Although the identity of the GI is unknown, phylogenetic and evolutionary analyses have indicated that S locus F-box like 1-3 (or S locus F-box with low allelic sequence polymorphism 1-3; SLFL1-3), which are encoded by a region of the Prunus genome linked to the S locus, are good GI candidates. Here, we examined the biochemical characteristics of SLFL1-3 to determine whether they have appropriate GI characteristics. Pull-down assays and quantitative expression analyses indicated that Prunus avium SLFL1-3 mainly formed a canonical SCF complex with PavSSK1 and PavCul1A. Binding assays with PavS(1,3,4,6)-RNases showed that PavSLFL1, PavSLFL2, and PavSLFL3 bound to PavS(3)-RNase, all PavS-RNases tested, and none of the PavS-RNases tested, respectively. Together, these results suggested that SLFL2 has the appropriate characteristics to be the GI in sweet cherry pollen, while SLFL1 may redundantly work with SLFL2 to detoxify all S-RNases. We discuss the possible roles of SLFL1-3 as the GI in the Prunus-specific S-RNase-based GSI mechanism.

[Investigation of radioactivity measurement of medical radioactive waste].

PubMed

Koizumi, Kiyoshi; Masuda, Kazutaka; Kusakabe, Kiyoko; Kinoshita, Fujimi; Kobayashi, Kazumi; Yamamoto, Tetsuo; Kanaya, Shinichi; Kida, Tetsuo; Yanagisawa, Masamichi; Iwanaga, Tetsuo; Ikebuchi, Hideharu; Kusama, Keiji; Namiki, Nobuo; Okuma, Hiroshi; Fujimura, Yoko; Horikoshi, Akiko; Tanaka, Mamoru

2004-11-01

To explore the possibility of which medical radioactive wastes could be disposed as general wastes after keeping them a certain period of time and confirming that their radioactivity reach a background level (BGL), we made a survey of these wastes in several nuclear medicine facilities. The radioactive wastes were collected for one week, packed in a box according to its half-life, and measured its radioactivity by scintillation survey meter with time. Some wastes could reach a BGL within 10 times of half-life, but 19% of the short half-life group (group 1) including 99mTc and 123I, and 8% of the middle half-life group (group 2) including 67Ga, (111)In, and 201Tl did not reach a BGL within 20 times of half-life. A reason for delaying the time of reaching a BGL might be partially attributed to high initial radiation dose rate or heavy package weight. However, mixing with the nuclides of longer half-life was estimated to be the biggest factor affecting this result. When disposing medical radioactive wastes as general wastes, it is necessary to avoid mixing with radionuclide of longer half-life and confirm that it reaches a BGL by actual measurement.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shull, H.E.

The objective of the project was to investigate the economic feasibility of converting potato waste to fuel alcohol. The source of potato starch was Troyer Farms Potato Chips. Experimental work was carried out at both the laboratory scale and the larger pilot scale batch operation at a decommissioned waste water treatment building on campus. The laboratory scale work was considerably more extensive than originally planned, resulting in a much improved scientific work. The pilot scale facility has been completed and operated successfully. In contrast, the analysis of the economic feasibility of commercial production has not yet been completed. The projectmore » was brought to a close with the successful demonstration of the fermentation and distillation using the large scale facilities described previously. Two batches of mash were cooked using the procedures established in support of the laboratory scale work. One of the batches was fermented using the optimum values of the seven controlled factors as predicted by the laboratory scale application of the Box-Wilson design. The other batch was fermented under conditions derived out of Mr. Rouse's interpretation of his long sequence of laboratory results. He was gratified to find that his commitment to the Box-Wilson experiments was justified. The productivity of the Box-Wilson design was greater. The difference between the performance of the two fermentors (one stirred, one not) has not been established yet. Both batches were then distilled together, demonstrating the satisfactory performance of the column still. 4 references.« less

Lead in the bone and soft tissues of box turtles caught near smelters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beresford, W.A.; Donovan, M.P.; Henninger, J.M.

1981-09-01

Three box turtles (Terrapene carolina) were caught by small streams in woodland south-east of Glover, MO and one, northeast of Bixby, MO: rural sites of primary lead smelters. Four other turtles were caught in a similar habitat, but distant from industry and main roads. Both groups shared the same food, water, and outside pen in Morgantown, WV. Lead content was assayed by atomic absorption spectrophotometry. Lead concentrations in the liver, kidney, skin, blood, and two long bone shafts were found to be significantly higher in the turtles captured near the smelters.

Development and Application of an Oversize Reusable DOT 7A Type A Overpack Container at the Y-12 National Security Complex - 13150

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tharp, Tim; Martin, David; Franco, Paul

2013-07-01

Waste Management personnel at the Y-12 National Security Complex (Y-12) are concluding a multi-year effort to dispose of a large backlog of low-level waste. Six containers presented a particularly difficult technical challenge in that they each contained large robust equipment (mostly salt baths) with elevated levels of highly enriched uranium (exceeding U.S. Department of Transportation (DOT) fissile-excepted quantities). The equipment was larger than the standard 1.2 m x 1.2 m x 1.8 m (4 ft x 4 ft x 6 ft) DOT Specification 7A Type A box and would have been very difficult to size-reduce because of several inches ofmore » steel plate (along with insulating block and concrete) in the equipment design. A critical breakthrough for the success of the project involved procuring and developing two oversize reusable DOT Specification 7A Type A (fissile tested) containers (referred to as the CTI Model 7AF-690-SC) that could be used as overpacks for the original boxes of equipment. The 7A Type A overpack containers are approximately 3.5 m long x 2.7 m wide x 2.8 m high (11.7 ft x 8.9 ft x 9.2 ft) with a maximum gross weight of 10,660 kg (23,500 lb) and a payload capacity of 6,804 kg (15,000 lbs). The boxes were designed and fabricated using a split cavity design that allowed the gasketed and bolted closure to lie along the horizontal centerline of the box. The central closure location in this design allows for strengthening of box corners that tend to be points of weakness or failure in 49CFR173.465 drop tests. By combining the split cavity design with large diameter tubing and diagonal cross bracing, drop test requirements of 49CFR173.465(1) and (2) were met and demonstrated through finite element analysis modeling. The development and use of this new container dramatically reduced the need for down-sizing the equipment and allowed the project to meet objectives within cost and schedule targets. (authors)« less

Optimization of Lactobacillus acidophilus cultivation using taro waste and evaluation of its biological activity.

PubMed

Hsieh, Shu-Chen; Liu, Jui-Ming; Pua, Xiao-Hui; Ting, Yuwen; Hsu, Ren-Jun; Cheng, Kuan-Chen

2016-03-01

In this study, taro waste (TW) was utilized for Lactobacillus acidophilus BCRC 14079 cultivation and the anti-tumor and immune-modulatory properties of heat-killed cells (HKCs), cytoplasmic fraction (CF), and exopolysaccharide (EPS) were evaluated. The optimum liquefaction enzyme dosage, temperature, and time determined by Box-Behnken design response surface methodology (BBD-RSM) were 9 mL/L of α-amylase, 79.2 °C, and 5 h of reaction, respectively. The optimum temperature and reaction time for saccharification were determined as 60 °C and 3 h. The optimum medium, CGMY1 medium, constitutes of TW hydrolysate containing 37 g/L of glucose, 25 g/L of corn gluten meal (CGM), and 1 g/L of yeast extract (YE). Results of MTT assay showed that HKCs and EPS from CGM medium exhibited the highest anti-proliferative in HT-29 (IC50 of HKCs, 467.25 μg/mL; EPS, 716.10 μg/mL) and in Caco-2 cells (IC50 of EPS, 741.60 μg/mL). Luciferase-based NF-ΚB and COX-2 systems indicated HKCs from CGM medium stimulated the highest expression of luciferin in both systems. The luciferase activities by using 100 and 500 μg/mL of HKCs from CGM were 24.30- and 45.83-fold in NF-ΚB system and 11.54- and 4.93-fold in COX-2 system higher than the control. In conclusion, this study demonstrated the potential of TW medium for L. acidophilus cultivation and the production of non-viable probiotics with enhanced biological activities.

Purification and DNA-binding properties of the cro-type regulatory repressor protein cng encoded by the Lactobacillus plantarum phage phi g1e.

PubMed

Kakikawa, M; Ohkubo, S; Sakate, T; Sayama, M; Taketo, A; Kodaira, K

2000-05-16

The putative repressor protein Cng (10kDa on an SDS gel) for the lytic pathway of Lactobacillus plantarum phage φg1e was purified using the Escherichia coli Pt7 system, and its DNA-binding ability for the seven operator-like sequences, the GATAC-boxes (Gb1 to Gb7), was investigated in vitro. In gel-shift assays, Cng selectively bound to the DNA fragments containing the GATAC-box(es). In addition, DNase I footprinting analysis with supercoiled DNA demonstrated that Cng can specifically cover about a 25bp region centered around each of the GATAC-boxes, although two boxes, Gb4 and Gb6, were only partially protected. Moreover, protein crosslinking experiments using glutaraldehyde suggested that Cng most likely functions as a dimer. On the other hand, the binding ability of Cpg for the GATAC-boxes in supercoiled DNA was also examined under the same conditions as in Cng; unlike Cng, Cpg covered Gb4 and Gb6 completely sufficiently as well as the other five boxes. Thus, the present and previous [Kakikawa et al., Gene 215 (1998) 371-379; 242 (2000) 155-166] results indicate a possibility that the two proteins Cng and Cpg selectively bind to the GATAC-boxes that act as operators, and can decide between the lytic or lysogenic pathways through repression of the promoter activity of P(R) as well as P(L).

Identification of She3 as an SCFGrr1 Substrate in Budding Yeast

PubMed Central

Wang, Ruiwen; Solomon, Mark J.

2012-01-01

The highly orchestrated progression of the cell cycle depends on the degradation of many regulatory proteins at different cell cycle stages. One of the key cell cycle ubiquitin ligases is the Skp1-cullin-F-box (SCF) complex. Acting in concert with the substrate-binding F-box protein Grr1, SCFGrr1 promotes the degradation of cell cycle regulators as well as various metabolic enzymes. Using a yeast two-hybrid assay with a Grr1 derivative as the bait, we identified She3, which is an adaptor protein in the asymmetric mRNA transport system, as a novel Grr1 substrate. We generated stabilized She3 mutants, which no longer bound to Grr1, and found that the degradation of She3 is not required for regulating asymmetric mRNA transport. However, She3 stabilization leads to slower growth compared to wild-type cells in a co-culture assay, demonstrating that the degradation of She3 by Grr1 is required for optimal cell growth. PMID:23144720

Profiles of embryonic nuclear protein binding to the proximal promoter region of the soybean β-conglycinin α subunit gene.

PubMed

Yoshino, M; Tsutsumi, K; Kanazawa, A

2015-01-01

β-Conglycinin, a major component of seed storage protein in soybean, comprises three subunits: α, α' and β. The expression of genes for these subunits is strictly controlled during embryogenesis. The proximal promoter region up to 245 bp upstream of the transcription start site of the α subunit gene sufficiently confers spatial and temporal control of transcription in embryos. Here, the binding profile of nuclear proteins in the proximal promoter region of the α subunit gene was analysed. DNase I footprinting analysis indicated binding of proteins to the RY element and DNA regions including box I, a region conserved in cognate gene promoters. An electrophoretic mobility shift assay (EMSA) using different portions of box I as a probe revealed that multiple portions of box I bind to nuclear proteins. In addition, an EMSA using nuclear proteins extracted from embryos at different developmental stages indicated that the levels of major DNA-protein complexes on box I increased during embryo maturation. These results are consistent with the notion that box I is important for the transcriptional control of seed storage protein genes. Furthermore, the present data suggest that nuclear proteins bind to novel motifs in box I including 5'-TCAATT-3' rather than to predicted cis-regulatory elements. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

Biological Control of the Chagas Disease Vector Triatoma infestans with the Entomopathogenic Fungus Beauveria bassiana Combined with an Aggregation Cue: Field, Laboratory and Mathematical Modeling Assessment

PubMed Central

Forlani, Lucas; Pedrini, Nicolás; Girotti, Juan R.; Mijailovsky, Sergio J.; Cardozo, Rubén M.; Gentile, Alberto G.; Hernández-Suárez, Carlos M.; Rabinovich, Jorge E.; Juárez, M. Patricia

2015-01-01

Background Current Chagas disease vector control strategies, based on chemical insecticide spraying, are growingly threatened by the emergence of pyrethroid-resistant Triatoma infestans populations in the Gran Chaco region of South America. Methodology and findings We have already shown that the entomopathogenic fungus Beauveria bassiana has the ability to breach the insect cuticle and is effective both against pyrethroid-susceptible and pyrethroid-resistant T. infestans, in laboratory as well as field assays. It is also known that T. infestans cuticle lipids play a major role as contact aggregation pheromones. We estimated the effectiveness of pheromone-based infection boxes containing B. bassiana spores to kill indoor bugs, and its effect on the vector population dynamics. Laboratory assays were performed to estimate the effect of fungal infection on female reproductive parameters. The effect of insect exuviae as an aggregation signal in the performance of the infection boxes was estimated both in the laboratory and in the field. We developed a stage-specific matrix model of T. infestans to describe the fungal infection effects on insect population dynamics, and to analyze the performance of the biopesticide device in vector biological control. Conclusions The pheromone-containing infective box is a promising new tool against indoor populations of this Chagas disease vector, with the number of boxes per house being the main driver of the reduction of the total domestic bug population. This ecologically safe approach is the first proven alternative to chemical insecticides in the control of T. infestans. The advantageous reduction in vector population by delayed-action fungal biopesticides in a contained environment is here shown supported by mathematical modeling. PMID:25969989

78 FR 64905 - Carriage of Conditionally Permitted Shale Gas Extraction Waste Water in Bulk

Federal Register 2010, 2011, 2012, 2013, 2014

2013-10-30

... the ``Keyword'' box. Click ``Search'' then click on the balloon shape in the ``Actions'' column. If... of 5 U.S.C. 552(a). The purpose of this notice is to announce the availability of the Coast Guard's... different wells. Upon reviewing comments received on this proposed policy letter, Coast Guard will issue the...

Using Dynamic Value Stream Mapping and Lean Accounting Box Scores to Support Lean Implementation

ERIC Educational Resources Information Center

Woehrle, Stephen L.; Abou-Shady, Louay

2010-01-01

Lean has proven to be an effective management philosophy for improving businesses in a competitive market by eliminating waste and improving operations. An impact of implementing lean projects is the rapid reduction in inventory levels, which gives management the false impression that profits are decreasing while workers on the shop floor observe…

40 CFR 62.15265 - How do I monitor the load of my municipal waste combustion unit?

Code of Federal Regulations, 2013 CFR

2013-07-01

... Mechanical Engineers (ASME PTC 4.1—1964): Test Code for Steam Generating Units, Power Test Code 4.1-1964... of Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (4) Design, construct...

40 CFR 62.15265 - How do I monitor the load of my municipal waste combustion unit?

Code of Federal Regulations, 2011 CFR

2011-07-01

... Mechanical Engineers (ASME PTC 4.1—1964): Test Code for Steam Generating Units, Power Test Code 4.1-1964... of Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (4) Design, construct...

40 CFR 62.15265 - How do I monitor the load of my municipal waste combustion unit?

Code of Federal Regulations, 2010 CFR

2010-07-01

... Mechanical Engineers (ASME PTC 4.1—1964): Test Code for Steam Generating Units, Power Test Code 4.1-1964... of Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (4) Design, construct...

40 CFR 62.15265 - How do I monitor the load of my municipal waste combustion unit?

Code of Federal Regulations, 2012 CFR

2012-07-01

... Mechanical Engineers (ASME PTC 4.1—1964): Test Code for Steam Generating Units, Power Test Code 4.1-1964... of Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (4) Design, construct...

40 CFR 62.15265 - How do I monitor the load of my municipal waste combustion unit?

Code of Federal Regulations, 2014 CFR

2014-07-01

... Mechanical Engineers (ASME PTC 4.1—1964): Test Code for Steam Generating Units, Power Test Code 4.1-1964... of Mechanical Engineers, Service Center, 22 Law Drive, Post Office Box 2900, Fairfield, NJ 07007. You....archives.gov/federal_register/code_of_federal_regulations/ibr_locations.html. (4) Design, construct...

76 FR 30700 - Agency Information Collection Activities; Submission to OMB for Review and Approval; Proposed...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-26

... prevention and waste management data, including recycling information, for such chemicals. 42 U.S.C. 13106..., install, and utilize technology and systems for the purposes of collecting, validating, and verifying... (Part I: Sec. 5, 5.1) with a check box that reads ``No U.S. Parent Company (for TRI Reporting purposes...

HEU Holdup Measurements on 321-M A-Lathe

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dewberry, R.A.

The Analytical Development Section of SRTC was requested by the Facilities Disposition Division (FDD) of the Savannah River Site to determine the holdup of enriched uranium in the 321-M facility as part of an overall deactivation project of the facility. The 321-M facility was used to fabricate enriched uranium fuel assemblies, lithium-aluminum target tubes, neptunium assemblies, and miscellaneous components for the production reactors. The results of the holdup assays are essential for determining compliance with the solid waste Waste Acceptance Criteria, Material Control and Accountability, and to meet criticality safety controls. Three measurement systems were used to determine highly enrichedmore » uranium (HEU) holdup. This report covers holdup measurements on the A-Lathe that was used to machine uranium-aluminum-alloy (U-Al). Our results indicated that the lathe contained more than the limits stated in the Waste Acceptance Criteria (WAC) for the solid waste E-Area Vaults. Thus the lathe was decontaminated three times and assayed four times in order to bring the amounts of uranium to an acceptable content. This report will discuss the methodology, Non-Destructive Assay (NDA) measurements, and results of the U-235 holdup on the lathe.« less

High-level waste tank farm set point document

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anthony, J.A. III

1995-01-15

Setpoints for nuclear safety-related instrumentation are required for actions determined by the design authorization basis. Minimum requirements need to be established for assuring that setpoints are established and held within specified limits. This document establishes the controlling methodology for changing setpoints of all classifications. The instrumentation under consideration involve the transfer, storage, and volume reduction of radioactive liquid waste in the F- and H-Area High-Level Radioactive Waste Tank Farms. The setpoint document will encompass the PROCESS AREA listed in the Safety Analysis Report (SAR) (DPSTSA-200-10 Sup 18) which includes the diversion box HDB-8 facility. In addition to the PROCESS AREASmore » listed in the SAR, Building 299-H and the Effluent Transfer Facility (ETF) are also included in the scope.« less

The help of simulation codes in designing waste assay systems using neutron measurement methods: Application to the alpha low level waste assay system PROMETHEE 6

NASA Astrophysics Data System (ADS)

Mariani, A.; Passard, C.; Jallu, F.; Toubon, H.

2003-11-01

The design of a specific nuclear assay system for a dedicated application begins with a phase of development, which relies on information from the literature or on knowledge resulting from experience, and on specific experimental verifications. The latter ones may require experimental devices which can be restricting in terms of deadline, cost and safety. One way generally chosen to bypass these difficulties is to use simulation codes to study particular aspects. This paper deals with the potentialities offered by the simulation in the case of a passive-active neutron (PAN) assay system for alpha low level waste characterization; this system has been carried out at the Nuclear Measurements Development Laboratory of the French Atomic Energy Commission. Due to the high number of parameters to be taken into account for its development, this is a particularly sophisticated example. Since the PAN assay system, called PROMETHEE (prompt epithermal and thermal interrogation experiment), must have a detection efficiency of more than 20% and preserve a high level of modularity for various applications, an improved version has been studied using the MCNP4 (Monte Carlo N-Particle) transport code. Parameters such as the dimensions of the assay system, of the cavity and of the detection blocks, and the thicknesses of the nuclear materials of neutronic interest have been optimised. Therefore, the number of necessary experiments was reduced.

Genetic and Genomic Insights into the Role of Benzoate-Catabolic Pathway Redundancy in Burkholderia xenovorans LB400†

PubMed Central

Denef, V. J.; Klappenbach, J. A.; Patrauchan, M. A.; Florizone, C.; Rodrigues, J. L. M.; Tsoi, T. V.; Verstraete, W.; Eltis, L. D.; Tiedje, J. M.

2006-01-01

Transcriptomic and proteomic analyses of Burkholderia xenovorans LB400, a potent polychlorinated biphenyl (PCB) degrader, have implicated growth substrate- and phase-dependent expression of three benzoate-catabolizing pathways: a catechol ortho cleavage (ben-cat) pathway and two benzoyl-coenzyme A pathways, encoded by gene clusters on the large chromosome (boxC) and the megaplasmid (boxM). To elucidate the significance of this apparent redundancy, we constructed mutants with deletions of the ben-cat pathway (the ΔbenABCD::kan mutant), the boxC pathway (the ΔboxABC::kan mutant), and both pathways (the ΔbenABCDΔ boxABC::kan mutant). All three mutants oxidized benzoate in resting-cell assays. However, the ΔbenABCD::kan and ΔbenABCD ΔboxABC::kan mutants grew at reduced rates on benzoate and displayed increased lag phases. By contrast, growth on succinate, on 4-hydroxybenzoate, and on biphenyl was unaffected. Microarray and proteomic analyses revealed that cells of the ΔbenABCD::kan mutant growing on benzoate expressed both box pathways. Overall, these results indicate that all three pathways catabolize benzoate. Deletion of benABCD abolished the ability of LB400 to grow using 3-chlorobenzoate. None of the benzoate pathways could degrade 2- or 4-chlorobenzoate, indicating that the pathway redundancy does not directly contribute to LB400's PCB-degrading capacities. Finally, an extensive sigmaE-regulated oxidative stress response not present in wild-type LB400 grown on benzoate was detected in these deletion mutants, supporting our earlier suggestion that the box pathways are preferentially active under reduced oxygen tension. Our data further substantiate the expansive network of tightly interconnected and complexly regulated aromatic degradation pathways in LB400. PMID:16391095

Max-E47, a Designed Minimalist Protein that Targets the E-Box DNA Site In Vivo and In Vitro

PubMed Central

Xu, Jing; Chen, Gang; De Jong, Antonia T.; Shahravan, S. Hesam; Shin, Jumi A.

2009-01-01

Max-E47 is a designed hybrid protein comprising the Max DNA-binding basic region and E47 HLH dimerization subdomain. In the yeast one-hybrid system (Y1H), Max-E47 shows strong transcriptional activation from the E-box site, 5'-CACGTG, targeted by the Myc/Max/Mad network of transcription factors; two mutants, Max-E47Y and Max-E47YF, activate more weakly from the E-box in the Y1H. Quantitative fluorescence anisotropy titrations to gain free energies of protein:DNA binding gave low nM Kd values for the native MaxbHLHZ, Max-E47, and the Y and YF mutants binding to the E-box site (14 nM, 15 nM, 9 nM, and 6 nM, respectively), with no detectable binding to a nonspecific control duplex. Because these minimalist, E-box-binding hybrids have no activation domain and no interactions with the c-MycbHLHZ, as shown by the yeast two-hybrid assay, they can potentially serve as dominant-negative inhibitors that suppress activation of E-box-responsive genes targeted by transcription factors including the c-Myc/Max complex. As proof-of-principle, we used our modified Y1H, which allows direct competition between two proteins vying for a DNA target, to show that Max-E47 effectively outcompetes the native MaxbHLHZ for the E-box; weaker competition is observed from the two mutants, consistent with Y1H results. These hybrids provide a minimalist scaffold for further exploration of the relationship between protein structure and DNA-binding function and may have applications as protein therapeutics or biochemical probes capable of targeting the E-box site. PMID:19449889

POZ domain transcription factor, FBI-1, represses transcription of ADH5/FDH by interacting with the zinc finger and interfering with DNA binding activity of Sp1.

PubMed

Lee, Dong-Kee; Suh, Dongchul; Edenberg, Howard J; Hur, Man-Wook

2002-07-26

The POZ domain is a protein-protein interaction motif that is found in many transcription factors, which are important for development, oncogenesis, apoptosis, and transcription repression. We cloned the POZ domain transcription factor, FBI-1, that recognizes the cis-element (bp -38 to -22) located just upstream of the core Sp1 binding sites (bp -22 to +22) of the ADH5/FDH minimal promoter (bp -38 to +61) in vitro and in vivo, as revealed by electrophoretic mobility shift assay and chromatin immunoprecipitation assay. The ADH5/FDH minimal promoter is potently repressed by the FBI-1. Glutathione S-transferase fusion protein pull-down showed that the POZ domains of FBI-1, Plzf, and Bcl-6 directly interact with the zinc finger DNA binding domain of Sp1. DNase I footprinting assays showed that the interaction prevents binding of Sp1 to the GC boxes of the ADH5/FDH promoter. Gal4-POZ domain fusions targeted proximal to the GC boxes repress transcription of the Gal4 upstream activator sequence-Sp1-adenovirus major late promoter. Our data suggest that POZ domain represses transcription by interacting with Sp1 zinc fingers and by interfering with the DNA binding activity of Sp1.

The bZIP transcription factor MdHY5 regulates anthocyanin accumulation and nitrate assimilation in apple.

PubMed

An, Jian-Ping; Qu, Feng-Jia; Yao, Ji-Fang; Wang, Xiao-Na; You, Chun-Xiang; Wang, Xiao-Fei; Hao, Yu-Jin

2017-01-01

The basic leucine zipper (bZIP) transcription factor HY5 plays a multifaceted role in plant growth and development. Here the apple MdHY5 gene was cloned based on its homology with Arabidopsis HY5 . Expression analysis demonstrated that MdHY5 transcription was induced by light and abscisic acid treatments. Electrophoretic mobility shift assays and transient expression assays subsequently showed that MdHY5 positively regulated both its own transcription and that of MdMYB10 by binding to E-box and G-box motifs, respectively. Furthermore, we obtained transgenic apple calli that overexpressed the MdHY5 gene, and apple calli coloration assays showed that MdHY5 promoted anthocyanin accumulation by regulating expression of the MdMYB10 gene and downstream anthocyanin biosynthesis genes. In addition, the transcript levels of a series of nitrate reductase genes and nitrate uptake genes in both wild-type and transgenic apple calli were detected. In association with increased nitrate reductase activities and nitrate contents, the results indicated that MdHY5 might be an important regulator in nutrient assimilation. Taken together, these results indicate that MdHY5 plays a vital role in anthocyanin accumulation and nitrate assimilation in apple.

Antiapoptotic Effect of Recombinant HMGB1 A-box Protein via Regulation of microRNA-21 in Myocardial Ischemia-Reperfusion Injury Model in Rats.

PubMed

Han, Qiang; Zhang, Hua-Yong; Zhong, Bei-Long; Zhang, Bing; Chen, Hua

2016-04-01

The ~80 amino acid A box DNA-binding domain of high mobility group box 1 (HMGB1) protein antagonizes proinflammatory responses during myocardial ischemia reperfusion (I/R) injury. The exact role of microRNA-21 (miR-21) is unknown, but its altered levels are evident in I/R injury. This study examined the roles of HMGB1 A-box and miR-21 in rat myocardial I/R injury model. Sixty Sprague-Dawley rats were randomly divided into six equal groups: (1) Sham; (2) I/R; (3) Ischemic postconditioning (IPost); (4) AntagomiR-21 post-treatment; (5) Recombinant HMGB1 A-box pretreatment; and (6) Recombinant HMGB1 A-box + antagomiR-21 post-treatment. Hemodynamic indexes, arrhythmia scores, ischemic area and infarct size, myocardial injury, and related parameters were studied. Expression of miR-21 was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to quantify apoptosis. Left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), maximal rate of pressure rise (+dp/dtmax), and decline (-dp/dtmax) showed clear reduction upon treatment with recombinant HMGB1 A-box. Arrhythmia was relieved and infarct area decreased in the group pretreated with recombinant HMGB1 A-box, compared with other groups. Circulating lactate dehydrogenase (LDH) and malondialdehyde (MDA) levels increased in response to irreversible cellular injury, while creatine kinase MB isoenzymes (CK-MB) and superoxide dismutase (SOD) activities were reduced in the I/R group, which was reversed following recombinant HMGB1 A-box treatment. Interestingly, pretreatment with recombinant HMGB1 A-box showed the most dramatic reductions in miR-21 levels, compared with other groups. Significantly reduced apoptotic index (AI) was seen in recombinant HMGB1 A-box pretreatment group and recombinant HMGB1 A-box + antagomiR-21 post-treatment group, with the former showing a more dramatic lowering in AI than the latter. Bax, caspase-8, and CHOP showed reduced expression, and Bcl-2 and p-AKT levels were upregulated in recombinant HMGB1 A-box pretreatment group. Thus, recombinant HMGB1 A-box treatment protects against I/R injury and the mechanisms may involve inhibition of miR-21 expression.

Measurement of radionuclides in waste packages

DOEpatents

Brodzinski, R.L.; Perkins, R.W.; Rieck, H.G.; Wogman, N.A.

1984-09-12

A method is described for non-destructively assaying the radionuclide content of solid waste in a sealed container by analysis of the waste's gamma-ray spectrum and neutron emissions. Some radionuclides are measured by characteristic photopeaks in the gamma-ray spectrum; transuranic nuclides are measured by neutron emission rate; other radionuclides are measured by correlation with those already measured.

Measurement of radionuclides in waste packages

DOEpatents

Brodzinski, Ronald L.; Perkins, Richard W.; Rieck, Henry G.; Wogman, Ned A.

1986-01-01

A method is described for non-destructively assaying the radionuclide content of solid waste in a sealed container by analysis of the waste's gamma-ray spectrum and neutron emissions. Some radionuclides are measured by characteristic photopeaks in the gamma-ray spectrum; transuranic nuclides are measured by neutron emission rate; other radionuclides are measured by correlation with those already measured.

Gateway BioBox: A Compact, Multi-Purpose Biological Hardware Suite for In Situ Experiments and Analyses in Deep Space

NASA Astrophysics Data System (ADS)

Smith, D. J.; Parra, M.; Lane, M.; Almeida, E. A.; Space Biosciences Research Branch

2018-02-01

A compilation of NASA's smallest biological hardware systems (plus 1-g gravity controls and ancillary sensors) that will allow for a wide range of specimen cultivation and analysis, from molecular measurements to broader cell and tissue assays.

HK022 Nun Requires Arginine-Rich Motif Residues Distinct from λ N

PubMed Central

Tawk, Caroline S.; Ghattas, Ingrid R.

2015-01-01

ABSTRACT Bacteriophage λ N protein binds boxB RNA hairpins in the nut (N utilization) sites of immediate early λ transcripts and interacts with host factors to suppress transcriptional termination at downstream terminators. In opposition to λ N, the Nun protein of HK022 binds the boxBs of coinfecting λ transcripts, interacts with a similar or identical set of host factors, and terminates transcription to suppress λ replication. Comparison of N-boxB and Nun-boxB nuclear magnetic resonance (NMR) structural models suggests similar interactions, though limited mutagenesis of Nun is available. Here, libraries of Nun's arginine-rich motif (ARM) were screened for the ability to exclude λ coinfection, and mutants were assayed for Nun termination with a boxB plasmid reporter system. Several Nun ARM residues appear to be immutable: Asp26, Arg28, Arg29, Arg32, Trp33, and Arg36. Asp26 and Trp33 appear to be unable to contact boxB and are not found at equivalent positions in λ N ARM. To understand if the requirement of Asp26, Trp33, and Arg36 indicated differences between HK022 Nun termination and λ N antitermination complexes, the same Nun libraries were fused to the activation domain of λ N and screened for clones able to complement N-deficient λ. Mutants were assayed for N antitermination. Surprisingly, Asp26 and Trp33 were still essential when Nun ARM was fused to N. Docking suggests that Nun ARM contacts a hydrophobic surface of the NusG carboxy-terminal domain containing residues necessary for Nun function. These findings indicate that Nun ARM relies on distinct contacts in its ternary complex and illustrate how protein-RNA recognition can evolve new regulatory functions. IMPORTANCE λ N protein interacts with host factors to allow λ nut-containing transcripts to elongate past termination signals. A competing bacteriophage, HK022, expresses Nun protein, which causes termination of λ nut transcripts. λ N and HK022 Nun use similar arginine-rich motifs (ARMs) to bind the same boxB RNAs in nut transcripts. Screening libraries of Nun ARM mutants, both in HK022 Nun and in a λ N fusion, revealed amino acids essential to Nun that could bind one or more host factors. Docking suggests that NusG, which is present in both Nun termination and N antitermination, is a plausible partner. These findings could help understand how transcription elongation is regulated and illustrate how subtle differences allow ARMs to evolve new regulatory functions. PMID:26350130

HK022 Nun Requires Arginine-Rich Motif Residues Distinct from λ N.

PubMed

Tawk, Caroline S; Ghattas, Ingrid R; Smith, Colin A

2015-11-01

Bacteriophage λ N protein binds boxB RNA hairpins in the nut (N utilization) sites of immediate early λ transcripts and interacts with host factors to suppress transcriptional termination at downstream terminators. In opposition to λ N, the Nun protein of HK022 binds the boxBs of coinfecting λ transcripts, interacts with a similar or identical set of host factors, and terminates transcription to suppress λ replication. Comparison of N-boxB and Nun-boxB nuclear magnetic resonance (NMR) structural models suggests similar interactions, though limited mutagenesis of Nun is available. Here, libraries of Nun's arginine-rich motif (ARM) were screened for the ability to exclude λ coinfection, and mutants were assayed for Nun termination with a boxB plasmid reporter system. Several Nun ARM residues appear to be immutable: Asp26, Arg28, Arg29, Arg32, Trp33, and Arg36. Asp26 and Trp33 appear to be unable to contact boxB and are not found at equivalent positions in λ N ARM. To understand if the requirement of Asp26, Trp33, and Arg36 indicated differences between HK022 Nun termination and λ N antitermination complexes, the same Nun libraries were fused to the activation domain of λ N and screened for clones able to complement N-deficient λ. Mutants were assayed for N antitermination. Surprisingly, Asp26 and Trp33 were still essential when Nun ARM was fused to N. Docking suggests that Nun ARM contacts a hydrophobic surface of the NusG carboxy-terminal domain containing residues necessary for Nun function. These findings indicate that Nun ARM relies on distinct contacts in its ternary complex and illustrate how protein-RNA recognition can evolve new regulatory functions. λ N protein interacts with host factors to allow λ nut-containing transcripts to elongate past termination signals. A competing bacteriophage, HK022, expresses Nun protein, which causes termination of λ nut transcripts. λ N and HK022 Nun use similar arginine-rich motifs (ARMs) to bind the same boxB RNAs in nut transcripts. Screening libraries of Nun ARM mutants, both in HK022 Nun and in a λ N fusion, revealed amino acids essential to Nun that could bind one or more host factors. Docking suggests that NusG, which is present in both Nun termination and N antitermination, is a plausible partner. These findings could help understand how transcription elongation is regulated and illustrate how subtle differences allow ARMs to evolve new regulatory functions. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Estimates for Pu-239 loadings in burial ground culverts based on fast/slow neutron measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Winn, W.G.; Hochel, R.C.; Hofstetter, K.J.

1989-08-15

This report provides guideline estimates for Pu-239 mass loadings in selected burial ground culverts. The relatively high recorded Pu-239 contents of these culverts have been appraised as suspect relative to criticality concerns, because they were assayed only with the solid waste monitor (SWM) per gamma-ray counting. After 1985, subsequent waste was also assayed with the neutron coincidence counter (NCC), and a comparison of the assay methods showed that the NCC generally yielded higher assays than the SWM. These higher NCC readings signaled a need to conduct non-destructive/non-intrusive nuclear interrogations of these culverts, and a technical team conducted scoping measurements tomore » illustrate potential assay methods based on neutron and/or gamma counting. A fast/slow neutron method has been developed to estimate the Pu-239 in the culverts. In addition, loading records include the SWM assays of all Pu-239 cuts of some of the culvert drums and these data are useful in estimating the corresponding NCC drum assays from NCC vs SWM data. Together, these methods yield predictions based on direct measurements and statistical inference.« less

Functional roles and efficiencies of the thioredoxin boxes of calcium-binding proteins 1 and 2 in protein folding.

PubMed Central

Kramer, B; Ferrari, D M; Klappa, P; Pöhlmann, N; Söling, H D

2001-01-01

The rat luminal endoplasmic-recticulum calcium-binding proteins 1 and 2 (CaBP1 and CaBP2 respectively) are members of the protein disulphide-isomerase (PDI) family. They contain two and three thioredoxin boxes (Cys-Gly-His-Cys) respectively and, like PDI, may be involved in the folding of nascent proteins. We demonstrate here that CaBP1, similar to PDI and CaBP2, can complement the lethal phenotype of the disrupted Saccharomyces cerevisiae PDI gene, provided that the natural C-terminal Lys-Asp-Glu-Leu sequence is replaced by His-Asp-Glu-Leu. Both the in vitro RNase AIII-re-activation assays and in vivo pro-(carboxypeptidase Y) processing assays using CaBP1 and CaBP2 thioredoxin (trx)-box mutants revealed that, whereas the three trx boxes in CaBP2 seem to be functionally equivalent, the first trx box of CaBP1 is significantly more active than the second trx box. Furthermore, only about 65% re-activation of denatured reduced RNase AIII could be obtained with CaBP1 or CaBP2 compared with PDI, and the yield of PDI-catalysed reactions was significantly reduced in the presence of either CaBP1 or CaBP2. In contrast with PDI, neither CaBP1 nor CaBP2 could catalyse the renaturation of denatured glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which is a redox-independent process, and neither protein had any effect on the PDI-catalysed refolding of GAPDH. Furthermore, although PDI can bind peptides via its b' domain, a property it shares with PDIp, the pancreas-specific PDI homologue, and although PDI can bind malfolded proteins such as 'scrambled' ribonuclease, no such interactions could be detected for CaBP2. We conclude that: (1) both CaBP2 and CaBP1 lack peptide-binding activity for GAPDH attributed to the C-terminal region of the a' domain of PDI; (2) CaBP2 lacks the general peptide-binding activity attributed to the b' domain of PDI; (3) interaction of CaBP2 with substrate (RNase AIII) is different from that of PDI and substrate; and (4) both CaBP2 and CaBP1 may promote oxidative folding by different kinetic pathways. PMID:11415439

CAD Instructor Designs Eco-Friendly Shed

ERIC Educational Resources Information Center

Schwendau, Mark

2013-01-01

Dissatisfied with the options offered by big box stores--and wanting to save some money and go as green as possible--the author puts his design and construction skills to good use. In this article, he shares how he designed and built an eco-friendly shed. He says he is very pleased with the results of working with his own design, reducing waste,…

FINISHING FABRICATED METAL PRODUCTS WITH ...

EPA Pesticide Factsheets

This report provides a technical and economic evaluation of a polyester powder coating system applied to the exterior and interior surfaces of metal boxes fabricated for the telephone and cable industries. This evaluation summarized many of the requirements and benefits of a clean technology that effectively eliminates the use of hazardous solvents and prevents the generation of volatile organic emissions and hazardous solid waste. publish information

Microencapsulation of Theobroma cacao L. waste extract: optimization using response surface methodology.

PubMed

Gabbay Alves, Taís Vanessa; Silva da Costa, Russany; Aliakbarian, Bahar; Casazza, Alessandro Alberto; Perego, Patrizia; Carréra Silva Júnior, José Otávio; Ribeiro Costa, Roseane Maria; Converti, Attilio

2017-03-01

The cocoa extract (Theobroma cacao L.) has a significant amount of polyphenols (TP) with potent antioxidant activity (AA). This study aims to optimise microencapsulation of the extract of cocoa waste using chitosan and maltodextrin. Microencapsulation tests were performed according to a Box-Behnken factorial design, and the results were evaluated by response surface methodology with temperature, maltodextrin concentration (MD) and extract flowrate (EF) as independent variables, and the fraction of encapsulated TP, TP encapsulation yield, AA, yield of drying and solubility index as responses. The optimum conditions were: inlet temperature of 170 °C, MD of 5% and EF of 2.5 mL/min. HPLC analysis identified epicatechin as the major component of both the extract and microparticles. TP release was faster at pH 3.5 than in water. These results as a whole suggest that microencapsulation was successful and the final product can be used as a nutrient source for aquatic animal feed. Highlights Microencapsulation is optimised according to a factorial design of the Box-Behnken type. Epicatechin is the major component of both the extract and microcapsules. The release of polyphenols from microcapsules is faster at pH 3.5 than in water.

Performance Assessment of Human and Cattle Associated Quantitative Real-time PCR Assays - slides

EPA Science Inventory

The presentation overview is (1) Single laboratory performance assessment of human- and cattle associated PCR assays and (2) A Field Study: Evaluation of two human fecal waste management practices in Ohio watershed.

Kinetic modelling of anaerobic hydrolysis of solid wastes, including disintegration processes

DOE Office of Scientific and Technical Information (OSTI.GOV)

García-Gen, Santiago; Sousbie, Philippe; Rangaraj, Ganesh

2015-01-15

Highlights: • Fractionation of solid wastes into readily and slowly biodegradable fractions. • Kinetic coefficients estimation from mono-digestion batch assays. • Validation of kinetic coefficients with a co-digestion continuous experiment. • Simulation of batch and continuous experiments with an ADM1-based model. - Abstract: A methodology to estimate disintegration and hydrolysis kinetic parameters of solid wastes and validate an ADM1-based anaerobic co-digestion model is presented. Kinetic parameters of the model were calibrated from batch reactor experiments treating individually fruit and vegetable wastes (among other residues) following a new protocol for batch tests. In addition, decoupled disintegration kinetics for readily and slowlymore » biodegradable fractions of solid wastes was considered. Calibrated parameters from batch assays of individual substrates were used to validate the model for a semi-continuous co-digestion operation treating simultaneously 5 fruit and vegetable wastes. The semi-continuous experiment was carried out in a lab-scale CSTR reactor for 15 weeks at organic loading rate ranging between 2.0 and 4.7 g VS/L d. The model (built in Matlab/Simulink) fit to a large extent the experimental results in both batch and semi-continuous mode and served as a powerful tool to simulate the digestion or co-digestion of solid wastes.« less

HMGB1 a-Box Reverses Brain Edema and Deterioration of Neurological Function in a Traumatic Brain Injury Mouse Model.

PubMed

Yang, Lijun; Wang, Feng; Yang, Liang; Yuan, Yunchao; Chen, Yan; Zhang, Gengshen; Fan, Zhenzeng

2018-01-01

Traumatic brain injury (TBI) is a complex neurological injury in young adults lacking effective treatment. Emerging evidences suggest that inflammation contributes to the secondary brain injury following TBI, including breakdown of the blood brain barrier (BBB), subsequent edema and neurological deterioration. High mobility group box-1 (HMGB1) has been identified as a key cytokine in the inflammation reaction following TBI. Here, we investigated the therapeutic efficacy of HMGB1 A-box fragment, an antagonist competing with full-length HMGB1 for receptor binding, against TBI. TBI was induced by controlled cortical impact (CCI) in adult male mice. HMGB1 A-box fragment was given intravenously at 2 mg/kg/day for 3 days after CCI. HMGB1 A-box-treated CCI mice were compared with saline-treated CCI mice and sham mice in terms of BBB disruption evaluated by Evan's blue extravasation, brain edema by brain water content, cell death by propidium iodide staining, inflammation by Western blot and ELISA assay for cytokine productions, as well as neurological functions by the modified Neurological Severity Score, wire grip and beam walking tests. HMGB1 A-box reversed brain damages in the mice following TBI. It significantly reduced brain edema by protecting integrity of the BBB, ameliorated cell degeneration, and decreased expression of pro-inflammatory cytokines released in injured brain after TBI. These cellular and molecular effects were accompanied by improved behavioral performance in TBI mice. Notably, HMGB1 A-box blocked IL-1β-induced HMGB1 release, and preferentially attenuated TLR4, Myd88 and P65 in astrocyte cultures. Our data suggest that HMGB1 is involved in CCI-induced TBI, which can be inhibited by HMGB1 A-box fragment. Therefore, HMGB1 A-box fragment may have therapeutic potential for the secondary brain damages in TBI. © 2018 The Author(s). Published by S. Karger AG, Basel.

bHLH106 Integrates Functions of Multiple Genes through Their G-Box to Confer Salt Tolerance on Arabidopsis.

PubMed

Ahmad, Aftab; Niwa, Yasuo; Goto, Shingo; Ogawa, Takeshi; Shimizu, Masanori; Suzuki, Akane; Kobayashi, Kyoko; Kobayashi, Hirokazu

2015-01-01

An activation-tagging methodology was applied to dedifferentiated calli of Arabidopsis to identify new genes involved in salt tolerance. This identified salt tolerant callus 8 (stc8) as a gene encoding the basic helix-loop-helix transcription factor bHLH106. bHLH106-knockout (KO) lines were more sensitive to NaCl, KCl, LiCl, ABA, and low temperatures than the wild-type. Back-transformation of the KO line rescued its phenotype, and over-expression (OX) of bHLH106 in differentiated plants exhibited tolerance to NaCl. Green fluorescent protein (GFP) fused with bHLH106 revealed that it was localized to the nucleus. Prepared bHLH106 protein was subjected to electrophoresis mobility shift assays against E-box sequences (5'-CANNTG-3'). The G-box sequence 5'-CACGTG-3' had the strongest interaction with bHLH106. bHLH106-OX lines were transcriptomically analyzed, and resultant up- and down-regulated genes selected on the criterion of presence of a G-box sequence. There were 198 genes positively regulated by bHLH106 and 36 genes negatively regulated; these genes possessed one or more G-box sequences in their promoter regions. Many of these genes are known to be involved in abiotic stress response. It is concluded that bHLH106 locates at a branching point in the abiotic stress response network by interacting directly to the G-box in genes conferring salt tolerance on plants.

SVP-like MADS-box protein from Carya cathayensis forms higher-order complexes.

PubMed

Wang, Jingjing; Hou, Chuanming; Huang, Jianqin; Wang, Zhengjia; Xu, Yingwu

2015-03-01

To properly regulate plant flowering time and construct floral pattern, MADS-domain containing transcription factors must form multimers including homo- and hetero-dimers. They are also active in forming hetero-higher-order complexes with three to five different molecules. However, it is not well known if a MADS-box protein can also form homo-higher-order complex. In this study a biochemical approach is utilized to provide insight into the complex formation for an SVP-like MADS-box protein cloned from hickory. The results indicated that the protein is a heterogeneous higher-order complex with the peak population containing over 20 monomers. Y2H verified the protein to form homo-complex in yeast cells. Western blot of the hickory floral bud sample revealed that the protein exists in higher-order polymers in native. Deletion assays indicated that the flexible C-terminal residues are mainly responsible for the higher-order polymer formation and the heterogeneity. Current results provide direct biochemical evidences for an active MADS-box protein to be a high order complex, much higher than a quartermeric polymer. Analysis suggests that a MADS-box subset may be able to self-assemble into large complexes, and thereby differentiate one subfamily from the other in a higher-order structural manner. Present result is a valuable supplement to the action of mechanism for MADS-box proteins in plant development. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Passive Neutron Non-Destructive Assay for Remediation of Radiological Waste at Hanford Burial Grounds- 13189

DOE Office of Scientific and Technical Information (OSTI.GOV)

Simpson, A.; Pitts, M.; Ludowise, J.D.

The Hanford burial grounds contains a broad spectrum of low activity radioactive wastes, transuranic (TRU) wastes, and hazardous wastes including fission products, byproduct material (thorium and uranium), plutonium and laboratory chemicals. A passive neutron non-destructive assay technique has been developed for characterization of shielded concreted drums exhumed from the burial grounds. This method facilitates the separation of low activity radiological waste containers from TRU waste containers exhumed from the burial grounds. Two identical total neutron counting systems have been deployed, each consisting of He-3 detectors surrounded by a polyethylene moderator. The counts are processed through a statistical filter that removesmore » outliers in order to suppress cosmic spallation events and electronic noise. Upon completion of processing, a 'GO / NO GO' signal is provided to the operator based on a threshold level equivalent to 0.5 grams of weapons grade plutonium in the container being evaluated. This approach allows instantaneous decisions to be made on how to proceed with the waste. The counting systems have been set up using initial on-site measurements (neutron emitting standards loaded into surrogate waste containers) combined with Monte Carlo modeling techniques. The benefit of this approach is to allow the systems to extend their measurement ranges, in terms of applicable matrix types and container sizes, with minimal interruption to the operations at the burial grounds. (authors)« less

High Efficiency Particulate Air (HEPA) Filter Generation, Characterization, and Disposal Experiences at the Oak Ridge National Laboratory

DOE Office of Scientific and Technical Information (OSTI.GOV)

Coffey, D. E.

2002-02-28

High Efficiency Particulate Air filtration is an essential component of the containment and ventilation systems supporting the research and development activities at the Oak Ridge National Laboratory. High Efficiency Particulate Air filters range in size from 7.6cm (3 inch) by 10.2 cm (4 inch) cylindrical shape filters to filter array assemblies up to 2.1 m (7 feet) high by 1.5 m (5 feet) wide. Spent filters are grouped by contaminates trapped in the filter media and become one of the components in the respective waste stream. Waste minimization and pollution prevention efforts are applied for both radiological and non-radiological applications.more » Radiological applications include laboratory hoods, glove boxes, and hot cells. High Efficiency Particulate Air filters also are generated from intake or pre-filtering applications, decontamination activities, and asbestos abatement applications. The disposal avenues include sanitary/industrial waste, Resource Conservation and Recovery Act and Toxic Substance Control Act, regulated waste, solid low-level waste, contact handled transuranic, and remote handled transuranic waste. This paper discusses characterization and operational experiences associated with the disposal of the spent filters across multiple applications.« less

APNEA/WIT system nondestructive assay capability evaluation plan for select accessibly stored INEL RWMC waste forms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Becker, G.K.

1997-01-01

Bio-Imaging Research Inc. (BIR) and Lockheed Martin Speciality Components (LMSC) are engaged in a Program Research and Development Agreement and a Rapid Commercialization Initiative with the Department of Energy, EM-50. The agreement required BIR and LMSC to develop a data interpretation method that merges nondestructive assay and nondestructive examination (NDA/NDE) data and information sufficient to establish compliance with applicable National TRU Program (Program) waste characterization requirements and associated quality assurance performance criteria. This effort required an objective demonstration of the BIR and LMSC waste characterization systems in their standalone and integrated configurations. The goal of the test plan is tomore » provide a mechanism from which evidence can be derived to substantiate nondestructive assay capability and utility statement for the BIT and LMSC systems. The plan must provide for the acquisition, compilation, and reporting of performance data thereby allowing external independent agencies a basis for an objective evaluation of the standalone BIR and LMSC measurement systems, WIT and APNEA respectively, as well as an expected performance resulting from appropriate integration of the two systems. The evaluation is to be structured such that a statement regarding select INEL RWMC waste forms can be made in terms of compliance with applicable Program requirements and criteria.« less

Lab-on-a-Chip-Based PCR-RFLP Assay for the Detection of Malayan Box Turtle (Cuora amboinensis) in the Food Chain and Traditional Chinese Medicines

PubMed Central

Asing; Ali, Md. Eaqub; Abd Hamid, Sharifah Bee; Hossain, M. A. Motalib; Mustafa, Shuhaimi; Kader, Md. Abdul; Zaidul, I. S. M.

2016-01-01

The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected turtle species, but it is a lucrative item in the illegal wildlife trade because of its great appeal as an exotic food item and in traditional medicine. Although several polymerase chain reaction (PCR) assays to identify MBT by various routes have been documented, their applicability for forensic authentication remains inconclusive due to the long length of the amplicon targets, which are easily broken down by natural decomposition, environmental stresses or physiochemical treatments during food processing. To address this research gap, we developed, for the first time, a species-specific PCR-restriction fragment length polymorphism (RFLP) assay with a very short target length (120 bp) to detect MBT in the food chain; this authentication ensured better security and reliability through molecular fingerprints. The PCR-amplified product was digested with Bfa1 endonuclease, and distinctive restriction fingerprints (72, 43 and 5 bp) for MBT were found upon separation in a microfluidic chip-based automated electrophoresis system, which enhances the resolution of short oligos. The chances of any false negative identifications were eliminated through the use of a universal endogenous control for eukaryotes, and the limit of detection was 0.0001 ng DNA or 0.01% of the meat under admixed states. Finally, the optimized PCR-RFLP assay was validated for the screening of raw and processed commercial meatballs, burgers and frankfurters, which are very popular in most countries. The optimized PCR-RFLP assay was further used to screen MBT materials in 153 traditional Chinese medicines of 17 different brands and 62 of them were found MBT positive; wherein the ingredients were not declared in product labels. Overall, the novel assay demonstrated sufficient merit for use in any forensic and/or archaeological authentication of MBT, even under a state of decomposition. PMID:27716792

Lab-on-a-Chip-Based PCR-RFLP Assay for the Detection of Malayan Box Turtle (Cuora amboinensis) in the Food Chain and Traditional Chinese Medicines.

PubMed

Asing; Ali, Md Eaqub; Abd Hamid, Sharifah Bee; Hossain, M A Motalib; Mustafa, Shuhaimi; Kader, Md Abdul; Zaidul, I S M

2016-01-01

The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected turtle species, but it is a lucrative item in the illegal wildlife trade because of its great appeal as an exotic food item and in traditional medicine. Although several polymerase chain reaction (PCR) assays to identify MBT by various routes have been documented, their applicability for forensic authentication remains inconclusive due to the long length of the amplicon targets, which are easily broken down by natural decomposition, environmental stresses or physiochemical treatments during food processing. To address this research gap, we developed, for the first time, a species-specific PCR-restriction fragment length polymorphism (RFLP) assay with a very short target length (120 bp) to detect MBT in the food chain; this authentication ensured better security and reliability through molecular fingerprints. The PCR-amplified product was digested with Bfa1 endonuclease, and distinctive restriction fingerprints (72, 43 and 5 bp) for MBT were found upon separation in a microfluidic chip-based automated electrophoresis system, which enhances the resolution of short oligos. The chances of any false negative identifications were eliminated through the use of a universal endogenous control for eukaryotes, and the limit of detection was 0.0001 ng DNA or 0.01% of the meat under admixed states. Finally, the optimized PCR-RFLP assay was validated for the screening of raw and processed commercial meatballs, burgers and frankfurters, which are very popular in most countries. The optimized PCR-RFLP assay was further used to screen MBT materials in 153 traditional Chinese medicines of 17 different brands and 62 of them were found MBT positive; wherein the ingredients were not declared in product labels. Overall, the novel assay demonstrated sufficient merit for use in any forensic and/or archaeological authentication of MBT, even under a state of decomposition.

LANL OPERATING EXPERIENCE WITH THE WAND AND HERCULES PROTOTYPE SYSTEMS

DOE Office of Scientific and Technical Information (OSTI.GOV)

K. M. GRUETZMACHER; C. L. FOXX; S. C. MYERS

2000-09-01

The Waste Assay for Nonradioactive Disposal (WAND) and the High Efficiency Radiation Counters for Ultimate Low Emission Sensitivity (HERCULES) prototype systems have been operating at Los Alamos National Laboratory's (LANL's) Solid Waste Operation's (SWO'S) non-destructive assay (NDA) building since 1997 and 1998, respectively. These systems are the cornerstone of the verification program for low-density Green is Clean (GIC) waste at the Laboratory. GIC waste includes all non-regulated waste generated in radiological controlled areas (RCAS) that has been actively segregated as clean (i.e., nonradioactive) through the use of waste generator acceptable knowledge (AK). The use of this methodology alters LANL's pastmore » practice of disposing of all room trash generated in nuclear facilities in radioactive waste landfills. Waste that is verified clean can be disposed of at the Los Alamos County Landfill. It is estimated that 50-90% of the low-density room trash from radioactive material handling areas at Los Alamos might be free of contamination. This approach avoids the high cost of disposal of clean waste at a radioactive waste landfill. It also reduces consumption of precious space in the radioactive waste landfill where disposal of this waste provides no benefit to the public or the environment. Preserving low level waste (LLW) disposal capacity for truly radioactive waste is critical in this era when expanding existing radioactive waste landfills or permitting new ones is resisted by regulators and stakeholders. This paper describes the operating experience with the WAND and HERCULES since they began operation at SWO. Waste for verification by the WAND system has been limited so far to waste from the Plutonium Facility and the Solid Waste Operations Facility. A total of461 ft3 (13.1 m3) of low-density shredded waste and paper have been verified clean by the WAND system. The HERCULES system has been used to verify waste from four Laboratory facilities. These are the Solid Waste Operations Facility, the TA-48 Chemistry Facility, the Shops Facility, and the Environmental Facility. A total of 3150 ft3 (89.3 m3) of low-density waste has been verified clean by the HERCULES system.« less

Identification and characterization of cell-specific enhancer elements for the mouse ETF/Tead2 gene.

PubMed

Tanoue, Y; Yasunami, M; Suzuki, K; Ohkubo, H

2001-12-21

We have identified and characterized by transient transfection assays the cell-specific 117-bp enhancer sequence in the first intron of the mouse ETF (Embryonic TEA domain-containing factor)/Tead2 gene required for transcriptional activation in ETF/Tead2 gene-expressing cells, such as P19 cells. The 117-bp enhancer contains one GC-rich sequence (5'-GGGGCGGGG-3'), termed the GC box, and two tandemly repeated GA-rich sequences (5'-GGGGGAGGGG-3'), termed the proximal and distal GA elements. Further analyses, including transfection studies and electrophoretic mobility shift assays using a series of deletion and mutation constructs, indicated that Sp1, a putative activator, may be required to predominate over its competition with another unknown putative repressor, termed the GA element-binding factor, for binding to both the GC box, which overlapped with the proximal GA element, and the distal GA element in the 117-bp sequence in order to achieve a full enhancer activity. We also discuss a possible mechanism underlying the cell-specific enhancer activity of the 117-bp sequence.

Application of Molecular Techniques To Elucidate the Influence of Cellulosic Waste on the Bacterial Community Structure at a Simulated Low-Level-Radioactive-Waste Site▿ †

PubMed Central

Field, Erin K.; D'Imperio, Seth; Miller, Amber R.; VanEngelen, Michael R.; Gerlach, Robin; Lee, Brady D.; Apel, William A.; Peyton, Brent M.

2010-01-01

Low-level-radioactive-waste (low-level-waste) sites, including those at various U.S. Department of Energy sites, frequently contain cellulosic waste in the form of paper towels, cardboard boxes, or wood contaminated with heavy metals and radionuclides such as chromium and uranium. To understand how the soil microbial community is influenced by the presence of cellulosic waste products, multiple soil samples were obtained from a nonradioactive model low-level-waste test pit at the Idaho National Laboratory. Samples were analyzed using 16S rRNA gene clone libraries and 16S rRNA gene microarray (PhyloChip) analyses. Both methods revealed changes in the bacterial community structure with depth. In all samples, the PhyloChip detected significantly more operational taxonomic units, and therefore relative diversity, than the clone libraries. Diversity indices suggest that diversity is lowest in the fill and fill-waste interface (FW) layers and greater in the wood waste and waste-clay interface layers. Principal-coordinate analysis and lineage-specific analysis determined that the Bacteroidetes and Actinobacteria phyla account for most of the significant differences observed between the layers. The decreased diversity in the FW layer and increased members of families containing known cellulose-degrading microorganisms suggest that the FW layer is an enrichment environment for these organisms. These results suggest that the presence of the cellulosic material significantly influences the bacterial community structure in a stratified soil system. PMID:20305022

Quantitative CrAssphage PCR Assays for Human Fecal ...

EPA Pesticide Factsheets

Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, crAssphage, was discovered by metagenomic data mining and reported to be abundant in and closely associated with human fecal waste. To confirm bioinformatic predictions, 384 primer sets were designed along the length of the crAssphage genome. Based upon initial screening, two novel crAssphage qPCR assays (CPQ_056 and CPQ_064) were designed and evaluated in reference fecal samples and water matrices. The assays exhibited high specificities (98.6%) when tested against a large animal fecal reference library and were highly abundant in raw sewage and sewage impacted water samples. In addition, CPQ_056 and CPQ_064 assay performance was compared to HF183/BacR287 and HumM2 methods in paired experiments. Findings confirm viral crAssphage qPCR assays perform at a similar level to well established bacterial human-associated fecal source identification technologies. These new viral based assays could become important water quality management and research tools. To inform the public.

An effect-directed strategy for characterizing emerging chemicals in food contact materials made from paper and board.

PubMed

Rosenmai, Anna Kjerstine; Bengtström, Linda; Taxvig, Camilla; Trier, Xenia; Petersen, Jens Højslev; Svingen, Terje; Binderup, Mona-Lise; Barbara Medea Alice, van Vugt-Lussenburg; Dybdahl, Marianne; Granby, Kit; Vinggaard, Anne Marie

2017-08-01

Food contact materials (FCM) are any type of item intended to come into contact with foods and thus represent a potential source for human exposure to chemicals. Regarding FCMs made of paper and board, information pertaining to their chemical constituents and the potential impacts on human health remains scarce, which hampers safety evaluation. We describe an effect-directed strategy to identify and characterize emerging chemicals in paper and board FCMs. Twenty FCMs were tested in eight reporter gene assays, including assays for the AR, ER, AhR, PPARγ, Nrf2 and p53, as well as mutagenicity. All FCMs exhibited activities in at least one assay. As proof-of-principle, FCM samples obtained from a sandwich wrapper and a pizza box were carried through a complete step-by-step multi-tiered approach. The pizza box exhibited ER activity, likely caused by the presence of bisphenol A, dibutyl phthalate, and benzylbutyl phthalate. The sandwich wrapper exhibited AR antagonism, likely caused by abietic acid and dehydroabietic acid. Migration studies confirmed that the active chemicals can transfer from FCMs to food simulants. In conclusion, we report an effect-directed strategy that can identify hazards posed by FCMs made from paper and board, including the identification of the chemical(s) responsible for the observed activity. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Neutron coincidence counting based on time interval analysis with one- and two-dimensional Rossi-alpha distributions: an application for passive neutron waste assay

NASA Astrophysics Data System (ADS)

Bruggeman, M.; Baeten, P.; De Boeck, W.; Carchon, R.

1996-02-01

Neutron coincidence counting is commonly used for the non-destructive assay of plutonium bearing waste or for safeguards verification measurements. A major drawback of conventional coincidence counting is related to the fact that a valid calibration is needed to convert a neutron coincidence count rate to a 240Pu equivalent mass ( 240Pu eq). In waste assay, calibrations are made for representative waste matrices and source distributions. The actual waste however may have quite different matrices and source distributions compared to the calibration samples. This often results in a bias of the assay result. This paper presents a new neutron multiplicity sensitive coincidence counting technique including an auto-calibration of the neutron detection efficiency. The coincidence counting principle is based on the recording of one- and two-dimensional Rossi-alpha distributions triggered respectively by pulse pairs and by pulse triplets. Rossi-alpha distributions allow an easy discrimination between real and accidental coincidences and are aimed at being measured by a PC-based fast time interval analyser. The Rossi-alpha distributions can be easily expressed in terms of a limited number of factorial moments of the neutron multiplicity distributions. The presented technique allows an unbiased measurement of the 240Pu eq mass. The presented theory—which will be indicated as Time Interval Analysis (TIA)—is complementary to Time Correlation Analysis (TCA) theories which were developed in the past, but is from the theoretical point of view much simpler and allows a straightforward calculation of deadtime corrections and error propagation. Analytical expressions are derived for the Rossi-alpha distributions as a function of the factorial moments of the efficiency dependent multiplicity distributions. The validity of the proposed theory is demonstrated and verified via Monte Carlo simulations of pulse trains and the subsequent analysis of the simulated data.

METHOD FOR MANUFACTURING LAMINATED SHEETS FOR PROTECTION AGAINST RADIOACTIVE WASTES, AND PROTECTING AND PACKAGING MEANS MANUFACTURED WITH THESE SHEETS; Papierfabrik Wilhemstal Wilhelm Ernst

DOE Office of Scientific and Technical Information (OSTI.GOV)

None

1959-07-15

A description is given of laminated sheet, consisting of a first layer of absorbing and preferably fibrous material (e.g., filter or blotting paper, or felt), a second layer of adhesive, impermeable, and hydrophobic material (e.g., wax, bitumen, a polyvinyl or polyacrylic compound, or a polyhydrocarbon), and a third (and fourth) layer of rigid material more or less impermeable to liquids (e.g., metal (aluminum), polyvinyl chloride, polyethylene, or cardboard). These sheets can be used for covering laboratory tables and walls, for radiation protection (manufacture of clothes, etc.), or for packaging radioactive waste (manufacture of boxes, bags, etc.). (NPO)

Enhancing anaerobic digestion of food waste through biochemical methane potential assays at different substrate: inoculum ratios.

PubMed

Hobbs, Shakira R; Landis, Amy E; Rittmann, Bruce E; Young, Michelle N; Parameswaran, Prathap

2018-01-01

Food waste has a high energy potential that can be converted into useful energy in the form of methane via anaerobic digestion. Biochemical Methane Potential assays (BMPs) were conducted to quantify the impacts on methane production of different ratios of food waste. Anaerobic digester sludge (ADS) was used as the inoculum, and BMPs were performed at food waste:inoculum ratios of 0.42, 1.42, and 3.0g chemical oxygen demand/g volatile solids (VS). The 1.42 ratio had the highest CH 4 -COD recovery: 90% of the initial total chemical oxygen demand (TCOD) was from food waste, followed by ratios 0.42 and 3.0 at 69% and 57%, respectively. Addition of food waste above 0.42 caused a lag time for CH 4 production that increased with higher ratios, which highlighted the negative impacts of overloading with food waste. The Gompertz equation was able to represent the results well, and it gave lag times of 0, 3.6 and 30days and maximum methane productions of 370, 910, and 1950mL for ratios 0.42, 1.42 and 3.0, respectively. While ratio 3.0 endured a long lag phase and low VSS destruction, ratio 1.42 achieved satisfactory results for all performance criteria. These results provide practical guidance on food-waste-to-inoculum ratios that can lead to optimizing methanogenic yield. Copyright © 2017 Elsevier Ltd. All rights reserved.

Hydrogen sulfide removal from sediment and water in box culverts/storm drains by iron-based granules.

PubMed

Sun, J L; Shang, C; Kikkert, G A

2013-01-01

A renewable granular iron-based technology for hydrogen sulfide removal from sediment and water in box culverts and storm drains is discussed. Iron granules, including granular ferric hydroxide (GFH), granular ferric oxide (GFO) and rusted waste iron crusts (RWIC) embedded in the sediment phase removed aqueous hydrogen sulfide formed from sedimentary biological sulfate reduction. The exhausted iron granules were exposed to dissolved oxygen and this regeneration process recovered the sulfide removal capacities of the granules. The recovery is likely attributable to the oxidation of the ferrous iron precipitates film and the formation of new reactive ferric iron surface sites on the iron granules and sand particles. GFH and RWIC showed larger sulfide removal capacities in the sediment phase than GFO, likely due to the less ordered crystal structures on their surfaces. This study demonstrates that the iron granules are able to remove hydrogen sulfide from sediment and water in box culverts and storm drains and they have the potential to be regenerated and reused by contacting with dissolved oxygen.

Dosage delivery of sensitive reagents enables glove-box-free synthesis

NASA Astrophysics Data System (ADS)

Sather, Aaron C.; Lee, Hong Geun; Colombe, James R.; Zhang, Anni; Buchwald, Stephen L.

2015-08-01

Contemporary organic chemists employ a broad range of catalytic and stoichiometric methods to construct molecules for applications in the material sciences, and as pharmaceuticals, agrochemicals, and sensors. The utility of a synthetic method may be greatly reduced if it relies on a glove box to enable the use of air- and moisture-sensitive reagents or catalysts. Furthermore, many synthetic chemistry laboratories have numerous containers of partially used reagents that have been spoiled by exposure to the ambient atmosphere. This is exceptionally wasteful from both an environmental and a cost perspective. Here we report an encapsulation method for stabilizing and storing air- and moisture-sensitive compounds. We demonstrate this approach in three contexts, by describing single-use capsules that contain all of the reagents (catalysts, ligands, and bases) necessary for the glove-box-free palladium-catalysed carbon-fluorine, carbon-nitrogen, and carbon-carbon bond-forming reactions. This strategy should reduce the number of error-prone, tedious and time-consuming weighing procedures required for such syntheses and should be applicable to a wide range of reagents, catalysts, and substrate combinations.

Cleanup Verification Package for the 118-H-6:2, 105-H Reactor Ancillary Support Areas, Below-Grade Structures, and Underlying Soils; the 118-H-6:3, 105-H Reactor Fuel Storage Basin and Underlying Soils; The 118-H-6:3 Fuel Storage Basin Deep Zone Side Slope Soils; the 100-H-9, 100-H-10, and 100-H-13 French Drains; the 100-H-11 and 100-H-12 Expansion Box French Drains; and the 100-H-14 and 100-H-31 Surface Contamination Zones

DOE Office of Scientific and Technical Information (OSTI.GOV)

M. J. Appel

2006-06-29

This cleanup verification package documents completion of removal actions for the 105-H Reactor Ancillary Support Areas, Below-Grade Structures, and Underlying Soils (subsite 118-H-6:2); 105-H Reactor Fuel Storage Basin and Underlying Soils (118-H-6:3); and Fuel Storage Basin Deep Zone Side Slope Soils. This CVP also documents remedial actions for the following seven additional waste sties: French Drain C (100-H-9), French Drain D (100-H-10), Expansion Box French Drain E (100-H-11), Expansion Box French Drain F (100-H-12), French Drain G (100-H-13), Surface Contamination Zone H (100-H-14), and the Polychlorinated Biphenyl Surface Contamination Zone (100-H-31).

Innovative Soft-Sided Waste Packaging System Implementation at a Small Department of Energy Environmental Restoration/Waste Management (ER/WM) Site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wolf, J.

2002-02-28

Weiss Associates (WA) performs a broad range of environmental restoration/waste management (ER/WM) activities for the U.S. Department of Energy (DOE) at the former Laboratory for Energy-Related Health Research (LEHR), University of California, Davis (UC Davis). Over the last three years, the LEHR ER/WM program transitioned from a baseline packaging system of steel, 2.7 cubic meter (3.5-cubic yard) B-25 boxes to a 7.0 cubic meter (9.1-cubic yard) soft-sided container (Lift Liner) system. The transition increased efficiencies in processing, packaging, and storage, and when combined with decreased procurement costs, achieved a $402,000 cost savings (Table I). Additional disposal costs between $128,600 andmore » $182,600 were avoided by minimizing void space. Future cost savings by the end of fiscal year 2003 are projected between $250,640 and $1,003,360.« less

Improvement of non-destructive fissile mass assays in α low-level waste drums: A matrix correction method based on neutron capture gamma-rays and a neutron generator

NASA Astrophysics Data System (ADS)

Jallu, F.; Loche, F.

2008-08-01

Within the framework of radioactive waste control, non-destructive assay (NDA) methods may be employed. The active neutron interrogation (ANI) method is now well-known and effective in quantifying low α-activity fissile masses (mainly 235U, 239Pu, 241Pu) with low densities, i.e. less than about 0.4, in radioactive waste drums of volumes up to 200 l. The PROMpt Epithermal and THErmal interrogation Experiment (PROMETHEE [F. Jallu, A. Mariani, C. Passard, A.-C. Raoux, H. Toubon, Alpha low level waste control: improvement of the PROMETHEE 6 assay system performances. Nucl. Technol. 153 (January) (2006); C. Passard, A. Mariani, F. Jallu, J. Romeyer-Dherber, H. Recroix, M. Rodriguez, J. Loridon, C. Denis, PROMETHEE: an alpha low level waste assay system using passive and active neutron measurement methods. Nucl. Technol. 140 (December) (2002) 303-314]) based on ANI has been under development since 1996 to reach the incinerating α low level waste (LLW) criterion of about 50 Bq[α] per gram of crude waste (≈50 μg Pu) in 118 l drums on the date the drums are conditioned. Difficulties arise when dealing with matrices containing neutron energy moderators such as H and neutron absorbents such as Cl. These components may have a great influence on the fissile mass deduced from the neutron signal measured by ANI. For example, the calibration coefficient measured in a 118 l drum containing a cellulose matrix (density d = 0.144 g cm -3) may be 50 times higher than that obtained in a poly-vinyl-chloride matrix ( d = 0.253 g cm -3). Without any information on the matrix, the fissile mass is often overestimated due to safety procedures and by considering the most disadvantageous calibration coefficient corresponding to the most absorbing and moderating calibration matrix. The work discussed in this paper was performed at the CEA Nuclear Measurement Laboratory in France. It concerns the development of a matrix effect correction method, which consists in identifying and quantifying the matrix components by using prompt gamma-rays following neutron capture. The method aims to refine the value of the adequate calibration coefficient used for ANI analysis. This paper presents the final results obtained for 118 l waste drums with low α-activity and low density. This paper discusses the experimental and modelling studies and describes the development of correction abacuses based on gamma-ray spectrometry signals.

Sequence-specific DNA binding by MYC/MAX to low-affinity non-E-box motifs.

PubMed

Allevato, Michael; Bolotin, Eugene; Grossman, Mark; Mane-Padros, Daniel; Sladek, Frances M; Martinez, Ernest

2017-01-01

The MYC oncoprotein regulates transcription of a large fraction of the genome as an obligatory heterodimer with the transcription factor MAX. The MYC:MAX heterodimer and MAX:MAX homodimer (hereafter MYC/MAX) bind Enhancer box (E-box) DNA elements (CANNTG) and have the greatest affinity for the canonical MYC E-box (CME) CACGTG. However, MYC:MAX also recognizes E-box variants and was reported to bind DNA in a "non-specific" fashion in vitro and in vivo. Here, in order to identify potential additional non-canonical binding sites for MYC/MAX, we employed high throughput in vitro protein-binding microarrays, along with electrophoretic mobility-shift assays and bioinformatic analyses of MYC-bound genomic loci in vivo. We identified all hexameric motifs preferentially bound by MYC/MAX in vitro, which include the low-affinity non-E-box sequence AACGTT, and found that the vast majority (87%) of MYC-bound genomic sites in a human B cell line contain at least one of the top 21 motifs bound by MYC:MAX in vitro. We further show that high MYC/MAX concentrations are needed for specific binding to the low-affinity sequence AACGTT in vitro and that elevated MYC levels in vivo more markedly increase the occupancy of AACGTT sites relative to CME sites, especially at distal intergenic and intragenic loci. Hence, MYC binds diverse DNA motifs with a broad range of affinities in a sequence-specific and dose-dependent manner, suggesting that MYC overexpression has more selective effects on the tumor transcriptome than previously thought.

Multiple circadian transcriptional elements cooperatively regulate cell-autonomous transcriptional oscillation of Period3, a mammalian clock gene.

PubMed

Matsumura, Ritsuko; Akashi, Makoto

2017-09-29

Cell-autonomous oscillation in clock gene expression drives circadian rhythms. The development of comprehensive analytical techniques, such as bioinformatics and ChIP-sequencing, has enabled the genome-wide identification of potential circadian transcriptional elements that regulate the transcriptional oscillation of clock genes. However, detailed analyses using traditional biochemical and molecular-biological approaches, such as binding and reporter assays, are still necessary to determine whether these potential circadian transcriptional elements are actually functional and how significantly they contribute to driving transcriptional oscillation. Here, we focused on the molecular mechanism of transcriptional oscillations in the mammalian clock gene Period3 ( Per3 ). The PER3 protein is essential for robust peripheral clocks and is a key component in circadian output processes. We found three E box-like elements located upstream of human Per3 transcription start sites that additively contributed to cell-autonomous transcriptional oscillation. However, we also found that Per3 is still expressed in a circadian manner when all three E box-like elements are functionally impaired. We noted that Per3 transcription was activated by the synergistic actions of two D box-like elements and the three E box-like elements, leading to a drastic increase in circadian amplitude. Interestingly, circadian expression of Per3 was completely disrupted only when all five transcriptional elements were functionally impaired. These results indicate that three E box-like and two D box-like elements cooperatively and redundantly regulate cell-autonomous transcriptional oscillation of Per3 . © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Health care industries: potential generators of genotoxic waste.

PubMed

Sharma, Pratibha; Kumar, Manish; Mathur, N; Singh, A; Bhatnagar, P; Sogani, M

2013-08-01

Health care waste includes all the waste generated by health care establishments, research facilities, and laboratories. This constitutes a variety of chemical substances, such as pharmaceuticals, radionuclides, solvents, and disinfectants. Recently, scientists and environmentalists have discovered that wastewater produced by hospitals possesses toxic properties due to various toxic chemicals and pharmaceuticals capable of causing environmental impacts and even lethal effects to organisms in aquatic ecosystems. Many of these compounds resist normal wastewater treatment and end up in surface waters. Besides aquatic organisms, humans can be exposed through drinking water produced from contaminated surface water. Indeed, some of the substances found in wastewaters are genotoxic and are suspected to be potential contributors to certain cancers. The aim of this study was to evaluate the genotoxic and cytotoxic potential of wastewaters from two hospitals and three clinical diagnostic centers located in Jaipur (Rajasthan State), India using the prokaryotic Salmonella mutagenicity assay (Ames assay) and the eukaryotic Saccharomyces cerevisiae respiration inhibition assay. In the Ames assay, untreated wastewaters from both of the health care sectors resulted in significantly increased numbers of revertant colonies up to 1,000-4,050 as measured by the Salmonella typhimurium TA98 and TA100 strains (with and without metabolic activation) after exposure to undiluted samples, which indicated the highly genotoxic nature of these wastewaters. Furthermore, both hospital and diagnostic samples were found to be highly cytotoxic. Effective concentrations at which 20 % (EC20) and 50 % (EC50) inhibition of the respiration rate of the cells occurred ranged between ~0.00 and 0.52 % and between 0.005 and 41.30 % (calculated with the help of the MS excel software XLSTAT 2012.1.01; Addinsoft), respectively, as determined by the S. cerevisiae assay. The results indicated that hospital wastewaters contain genotoxic and cytotoxic components. In addition, diagnostic centers also represent small but significant sources of genotoxic and cytotoxic wastes.

Overexpression of a novel MADS-box gene SlFYFL delays senescence, fruit ripening and abscission in tomato

PubMed Central

Xie, Qiaoli; Hu, Zongli; Zhu, Zhiguo; Dong, Tingting; Zhao, Zhiping; Cui, Baolu; Chen, Guoping

2014-01-01

MADS-domain proteins are important transcription factors involved in many biological processes of plants. In our study, a tomato MADS-box gene, SlFYFL, was isolated. SlFYFL is expressed in all tissues of tomato and significantly higher in mature leave, fruit of different stages, AZ (abscission zone) and sepal. Delayed leaf senescence and fruit ripening, increased storability and longer sepals were observed in 35S:FYFL tomato. The accumulation of carotenoid was reduced, and ethylene content, ethylene biosynthetic and responsive genes were down-regulated in 35S:FYFL fruits. Abscission zone (AZ) did not form normally and abscission zone development related genes were declined in AZs of 35S:FYFL plants. Yeast two-hybrid assay revealed that SlFYFL protein could interact with SlMADS-RIN, SlMADS1 and SlJOINTLESS, respectively. These results suggest that overexpression of SlFYFL regulate fruit ripening and development of AZ via interactions with the ripening and abscission zone-related MADS box proteins. PMID:24621662

Varicella-zoster virus (VZV) origin of DNA replication oriS influences origin-dependent DNA replication and flanking gene transcription.

PubMed

Khalil, Mohamed I; Sommer, Marvin H; Hay, John; Ruyechan, William T; Arvin, Ann M

2015-07-01

The VZV genome has two origins of DNA replication (oriS), each of which consists of an AT-rich sequence and three origin binding protein (OBP) sites called Box A, C and B. In these experiments, the mutation in the core sequence CGC of the Box A and C not only inhibited DNA replication but also inhibited both ORF62 and ORF63 expression in reporter gene assays. In contrast the Box B mutation did not influence DNA replication or flanking gene transcription. These results suggest that efficient DNA replication enhances ORF62 and ORF63 transcription. Recombinant viruses carrying these mutations in both sites and one with a deletion of the whole oriS were constructed. Surprisingly, the recombinant virus lacking both copies of oriS retained the capacity to replicate in melanoma and HELF cells suggesting that VZV has another origin of DNA replication. Copyright © 2015 Elsevier Inc. All rights reserved.

Cost assessment of the automated VERSANT 440 Molecular System versus the semi-automated System 340 bDNA Analyzer platforms.

PubMed

Elbeik, Tarek; Loftus, Richard A; Beringer, Scott

2007-11-01

Labor, supply and waste were evaluated for HIV-1 and HCV bDNA on the semi-automated System 340 bDNA Analyzer and the automated VERSANT 440 Molecular System (V440). HIV-1 sample processing was evaluated using a 24- and 48-position centrifuge rotor. Vigilance time (hands-on manipulations plus incubation time except initial target hybridization) and disposables were approximately 37 and 12% lower for HIV-1, and 64 and 31% lower for HCV bDNA, respectively, with V440. Biohazardous solid waste was approximately twofold lower for both assays and other waste types were the same for either assay on both platforms. HIV-1 sample processing vigilance time for the 48-position rotor was reduced by 2 h. V440 provides cost savings and improved workflow.

Upregulation of MiR-212 Inhibits Migration and Tumorigenicity and Inactivates Wnt/β-Catenin Signaling in Human Hepatocellular Carcinoma.

PubMed

Jia, Pengbo; Wei, Guangbing; Zhou, Cancan; Gao, Qi; Wu, Yunhua; Sun, Xuejun; Li, Xuqi

2018-01-01

MicroRNAs are involved in hepatocellular carcinoma metastasis, a principal cause of hepatocellular carcinoma-related death in patients worldwide. MiR-212 is a microRNA that has been identified in several types of cancers and is postulated to influence cell signaling and subsequent malignant pathogenesis. Despite emerging reports suggesting that miR-212 plays a significant role in the onset, progression, and migration of these types of malignant tumors, its involvement in the development of hepatocellular carcinoma has not been fully elucidated. Quantitative reverse transcription polymerase chain reaction, wound healing, transwell migration and invasion assays, Western blotting, and xenograft tumor growth models were performed to test the expression levels and functions of miR-212 in hepatocellular carcinoma. Luciferase reporter assay, quantitative reverse transcription polymerase chain reaction, Western blotting, and immunohistochemistry were used to identify and verify the target of miR-212. In this study, we identify significant repression of miR-212 in hepatocellular carcinoma and demonstrate that overexpression of miR-212 inhibits the migration of hepatocellular carcinoma cells in vitro and in vivo. Furthermore, we identify forkhead box M1, whose expression is inversely related to that of miR-212, as a direct target of miR-212. Additionally, reexpression of forkhead box M1 rescues the miR-212-mediated inhibition of cell migration. We observed that inhibition of miR-212 activates forkhead box M1 but inhibits the Wnt/β-catenin pathway by suppressing Wnt, LEF-1, c-Myc, and nuclear β-catenin. Finally, in vivo studies confirmed the inhibitory effect of miR-212 on hepatocellular carcinoma growth. Our present findings indicate that miR-212 is a potential prognostic biomarker of hepatocellular carcinoma and that the miR-212/forkhead box M1 regulatory axis may represent a new therapeutic objective for hepatocellular carcinoma treatment.

Engineering study of 50 miscellaneous inactive underground radioactive waste tanks located at the Hanford Site, Washington

DOE Office of Scientific and Technical Information (OSTI.GOV)

Freeman-Pollard, J.R.

1994-03-02

This engineering study addresses 50 inactive underground radioactive waste tanks. The tanks were formerly used for the following functions associated with plutonium and uranium separations and waste management activities in the 200 East and 200 West Areas of the Hanford Site: settling solids prior to disposal of supernatant in cribs and a reverse well; neutralizing acidic process wastes prior to crib disposal; receipt and processing of single-shell tank (SST) waste for uranium recovery operations; catch tanks to collect water that intruded into diversion boxes and transfer pipeline encasements and any leakage that occurred during waste transfer operations; and waste handlingmore » and process experimentation. Most of these tanks have not been in use for many years. Several projects have, been planned and implemented since the 1970`s and through 1985 to remove waste and interim isolate or interim stabilize many of the tanks. Some tanks have been filled with grout within the past several years. Responsibility for final closure and/or remediation of these tanks is currently assigned to several programs including Tank Waste Remediation Systems (TWRS), Environmental Restoration and Remedial Action (ERRA), and Decommissioning and Resource Conservation and Recovery Act (RCRA) Closure (D&RCP). Some are under facility landlord responsibility for maintenance and surveillance (i.e. Plutonium Uranium Extraction [PUREX]). However, most of the tanks are not currently included in any active monitoring or surveillance program.« less

Open Source Drug Discovery with the Malaria Box Compound Collection for Neglected Diseases and Beyond

PubMed Central

Van Voorhis, Wesley C.; Adams, John H.; Adelfio, Roberto; Ahyong, Vida; Akabas, Myles H.; Alano, Pietro; Alday, Aintzane; Alemán Resto, Yesmalie; Alsibaee, Aishah; Alzualde, Ainhoa; Andrews, Katherine T.; Avery, Simon V.; Avery, Vicky M.; Ayong, Lawrence; Baker, Mark; Baker, Stephen; Ben Mamoun, Choukri; Bhatia, Sangeeta; Bickle, Quentin; Bounaadja, Lotfi; Bowling, Tana; Bosch, Jürgen; Boucher, Lauren E.; Boyom, Fabrice F.; Brea, Jose; Brennan, Marian; Burton, Audrey; Caffrey, Conor R.; Camarda, Grazia; Carrasquilla, Manuela; Carter, Dee; Belen Cassera, Maria; Chih-Chien Cheng, Ken; Chindaudomsate, Worathad; Chubb, Anthony; Colon, Beatrice L.; Colón-López, Daisy D.; Corbett, Yolanda; Crowther, Gregory J.; Cowan, Noemi; D’Alessandro, Sarah; Le Dang, Na; Delves, Michael; Du, Alan Y.; Duffy, Sandra; Abd El-Salam El-Sayed, Shimaa; Ferdig, Michael T.; Fernández Robledo, José A.; Fidock, David A.; Florent, Isabelle; Fokou, Patrick V. T.; Galstian, Ani; Gamo, Francisco Javier; Gold, Ben; Golub, Todd; Goldgof, Gregory M.; Guha, Rajarshi; Guiguemde, W. Armand; Gural, Nil; Guy, R. Kiplin; Hansen, Michael A. E.; Hanson, Kirsten K.; Hemphill, Andrew; Hooft van Huijsduijnen, Rob; Horii, Takaaki; Horrocks, Paul; Hughes, Tyler B.; Huston, Christopher; Igarashi, Ikuo; Ingram-Sieber, Katrin; Itoe, Maurice A.; Jadhav, Ajit; Naranuntarat Jensen, Amornrat; Jensen, Laran T.; Jiang, Rays H. Y.; Kaiser, Annette; Keiser, Jennifer; Ketas, Thomas; Kicka, Sebastien; Kim, Sunyoung; Kirk, Kiaran; Kumar, Vidya P.; Kyle, Dennis E.; Lafuente, Maria Jose; Landfear, Scott; Lee, Nathan; Lee, Sukjun; Lehane, Adele M.; Li, Fengwu; Little, David; Liu, Liqiong; Llinás, Manuel; Loza, Maria I.; Lubar, Aristea; Lucantoni, Leonardo; Lucet, Isabelle; Maes, Louis; Mancama, Dalu; Mansour, Nuha R.; March, Sandra; McGowan, Sheena; Medina Vera, Iset; Meister, Stephan; Mercer, Luke; Mestres, Jordi; Mfopa, Alvine N.; Misra, Raj N.; Moon, Seunghyun; Moore, John P.; Morais Rodrigues da Costa, Francielly; Müller, Joachim; Muriana, Arantza; Nakazawa Hewitt, Stephen; Nare, Bakela; Nathan, Carl; Narraidoo, Nathalie; Nawaratna, Sujeevi; Ojo, Kayode K.; Ortiz, Diana; Panic, Gordana; Papadatos, George; Parapini, Silvia; Patra, Kailash; Pham, Ngoc; Prats, Sarah; Plouffe, David M.; Poulsen, Sally-Ann; Pradhan, Anupam; Quevedo, Celia; Quinn, Ronald J.; Rice, Christopher A.; Abdo Rizk, Mohamed; Ruecker, Andrea; St. Onge, Robert; Salgado Ferreira, Rafaela; Samra, Jasmeet; Robinett, Natalie G.; Schlecht, Ulrich; Schmitt, Marjorie; Silva Villela, Filipe; Silvestrini, Francesco; Sinden, Robert; Smith, Dennis A.; Soldati, Thierry; Spitzmüller, Andreas; Stamm, Serge Maximilian; Sullivan, David J.; Sullivan, William; Suresh, Sundari; Suzuki, Brian M.; Suzuki, Yo; Swamidass, S. Joshua; Taramelli, Donatella; Tchokouaha, Lauve R. Y.; Theron, Anjo; Thomas, David; Tonissen, Kathryn F.; Townson, Simon; Tripathi, Abhai K.; Trofimov, Valentin; Udenze, Kenneth O.; Ullah, Imran; Vallieres, Cindy; Vigil, Edgar; Vinetz, Joseph M.; Voong Vinh, Phat; Vu, Hoan; Watanabe, Nao-aki; Weatherby, Kate; White, Pamela M.; Wilks, Andrew F.; Winzeler, Elizabeth A.; Wojcik, Edward; Wree, Melanie; Wu, Wesley; Yokoyama, Naoaki; Zollo, Paul H. A.; Abla, Nada; Blasco, Benjamin; Burrows, Jeremy; Laleu, Benoît; Leroy, Didier; Spangenberg, Thomas; Wells, Timothy; Willis, Paul A.

2016-01-01

A major cause of the paucity of new starting points for drug discovery is the lack of interaction between academia and industry. Much of the global resource in biology is present in universities, whereas the focus of medicinal chemistry is still largely within industry. Open source drug discovery, with sharing of information, is clearly a first step towards overcoming this gap. But the interface could especially be bridged through a scale-up of open sharing of physical compounds, which would accelerate the finding of new starting points for drug discovery. The Medicines for Malaria Venture Malaria Box is a collection of over 400 compounds representing families of structures identified in phenotypic screens of pharmaceutical and academic libraries against the Plasmodium falciparum malaria parasite. The set has now been distributed to almost 200 research groups globally in the last two years, with the only stipulation that information from the screens is deposited in the public domain. This paper reports for the first time on 236 screens that have been carried out against the Malaria Box and compares these results with 55 assays that were previously published, in a format that allows a meta-analysis of the combined dataset. The combined biochemical and cellular assays presented here suggest mechanisms of action for 135 (34%) of the compounds active in killing multiple life-cycle stages of the malaria parasite, including asexual blood, liver, gametocyte, gametes and insect ookinete stages. In addition, many compounds demonstrated activity against other pathogens, showing hits in assays with 16 protozoa, 7 helminths, 9 bacterial and mycobacterial species, the dengue fever mosquito vector, and the NCI60 human cancer cell line panel of 60 human tumor cell lines. Toxicological, pharmacokinetic and metabolic properties were collected on all the compounds, assisting in the selection of the most promising candidates for murine proof-of-concept experiments and medicinal chemistry programs. The data for all of these assays are presented and analyzed to show how outstanding leads for many indications can be selected. These results reveal the immense potential for translating the dispersed expertise in biological assays involving human pathogens into drug discovery starting points, by providing open access to new families of molecules, and emphasize how a small additional investment made to help acquire and distribute compounds, and sharing the data, can catalyze drug discovery for dozens of different indications. Another lesson is that when multiple screens from different groups are run on the same library, results can be integrated quickly to select the most valuable starting points for subsequent medicinal chemistry efforts. PMID:27467575

Open Source Drug Discovery with the Malaria Box Compound Collection for Neglected Diseases and Beyond.

PubMed

Van Voorhis, Wesley C; Adams, John H; Adelfio, Roberto; Ahyong, Vida; Akabas, Myles H; Alano, Pietro; Alday, Aintzane; Alemán Resto, Yesmalie; Alsibaee, Aishah; Alzualde, Ainhoa; Andrews, Katherine T; Avery, Simon V; Avery, Vicky M; Ayong, Lawrence; Baker, Mark; Baker, Stephen; Ben Mamoun, Choukri; Bhatia, Sangeeta; Bickle, Quentin; Bounaadja, Lotfi; Bowling, Tana; Bosch, Jürgen; Boucher, Lauren E; Boyom, Fabrice F; Brea, Jose; Brennan, Marian; Burton, Audrey; Caffrey, Conor R; Camarda, Grazia; Carrasquilla, Manuela; Carter, Dee; Belen Cassera, Maria; Chih-Chien Cheng, Ken; Chindaudomsate, Worathad; Chubb, Anthony; Colon, Beatrice L; Colón-López, Daisy D; Corbett, Yolanda; Crowther, Gregory J; Cowan, Noemi; D'Alessandro, Sarah; Le Dang, Na; Delves, Michael; DeRisi, Joseph L; Du, Alan Y; Duffy, Sandra; Abd El-Salam El-Sayed, Shimaa; Ferdig, Michael T; Fernández Robledo, José A; Fidock, David A; Florent, Isabelle; Fokou, Patrick V T; Galstian, Ani; Gamo, Francisco Javier; Gokool, Suzanne; Gold, Ben; Golub, Todd; Goldgof, Gregory M; Guha, Rajarshi; Guiguemde, W Armand; Gural, Nil; Guy, R Kiplin; Hansen, Michael A E; Hanson, Kirsten K; Hemphill, Andrew; Hooft van Huijsduijnen, Rob; Horii, Takaaki; Horrocks, Paul; Hughes, Tyler B; Huston, Christopher; Igarashi, Ikuo; Ingram-Sieber, Katrin; Itoe, Maurice A; Jadhav, Ajit; Naranuntarat Jensen, Amornrat; Jensen, Laran T; Jiang, Rays H Y; Kaiser, Annette; Keiser, Jennifer; Ketas, Thomas; Kicka, Sebastien; Kim, Sunyoung; Kirk, Kiaran; Kumar, Vidya P; Kyle, Dennis E; Lafuente, Maria Jose; Landfear, Scott; Lee, Nathan; Lee, Sukjun; Lehane, Adele M; Li, Fengwu; Little, David; Liu, Liqiong; Llinás, Manuel; Loza, Maria I; Lubar, Aristea; Lucantoni, Leonardo; Lucet, Isabelle; Maes, Louis; Mancama, Dalu; Mansour, Nuha R; March, Sandra; McGowan, Sheena; Medina Vera, Iset; Meister, Stephan; Mercer, Luke; Mestres, Jordi; Mfopa, Alvine N; Misra, Raj N; Moon, Seunghyun; Moore, John P; Morais Rodrigues da Costa, Francielly; Müller, Joachim; Muriana, Arantza; Nakazawa Hewitt, Stephen; Nare, Bakela; Nathan, Carl; Narraidoo, Nathalie; Nawaratna, Sujeevi; Ojo, Kayode K; Ortiz, Diana; Panic, Gordana; Papadatos, George; Parapini, Silvia; Patra, Kailash; Pham, Ngoc; Prats, Sarah; Plouffe, David M; Poulsen, Sally-Ann; Pradhan, Anupam; Quevedo, Celia; Quinn, Ronald J; Rice, Christopher A; Abdo Rizk, Mohamed; Ruecker, Andrea; St Onge, Robert; Salgado Ferreira, Rafaela; Samra, Jasmeet; Robinett, Natalie G; Schlecht, Ulrich; Schmitt, Marjorie; Silva Villela, Filipe; Silvestrini, Francesco; Sinden, Robert; Smith, Dennis A; Soldati, Thierry; Spitzmüller, Andreas; Stamm, Serge Maximilian; Sullivan, David J; Sullivan, William; Suresh, Sundari; Suzuki, Brian M; Suzuki, Yo; Swamidass, S Joshua; Taramelli, Donatella; Tchokouaha, Lauve R Y; Theron, Anjo; Thomas, David; Tonissen, Kathryn F; Townson, Simon; Tripathi, Abhai K; Trofimov, Valentin; Udenze, Kenneth O; Ullah, Imran; Vallieres, Cindy; Vigil, Edgar; Vinetz, Joseph M; Voong Vinh, Phat; Vu, Hoan; Watanabe, Nao-Aki; Weatherby, Kate; White, Pamela M; Wilks, Andrew F; Winzeler, Elizabeth A; Wojcik, Edward; Wree, Melanie; Wu, Wesley; Yokoyama, Naoaki; Zollo, Paul H A; Abla, Nada; Blasco, Benjamin; Burrows, Jeremy; Laleu, Benoît; Leroy, Didier; Spangenberg, Thomas; Wells, Timothy; Willis, Paul A

2016-07-01

A major cause of the paucity of new starting points for drug discovery is the lack of interaction between academia and industry. Much of the global resource in biology is present in universities, whereas the focus of medicinal chemistry is still largely within industry. Open source drug discovery, with sharing of information, is clearly a first step towards overcoming this gap. But the interface could especially be bridged through a scale-up of open sharing of physical compounds, which would accelerate the finding of new starting points for drug discovery. The Medicines for Malaria Venture Malaria Box is a collection of over 400 compounds representing families of structures identified in phenotypic screens of pharmaceutical and academic libraries against the Plasmodium falciparum malaria parasite. The set has now been distributed to almost 200 research groups globally in the last two years, with the only stipulation that information from the screens is deposited in the public domain. This paper reports for the first time on 236 screens that have been carried out against the Malaria Box and compares these results with 55 assays that were previously published, in a format that allows a meta-analysis of the combined dataset. The combined biochemical and cellular assays presented here suggest mechanisms of action for 135 (34%) of the compounds active in killing multiple life-cycle stages of the malaria parasite, including asexual blood, liver, gametocyte, gametes and insect ookinete stages. In addition, many compounds demonstrated activity against other pathogens, showing hits in assays with 16 protozoa, 7 helminths, 9 bacterial and mycobacterial species, the dengue fever mosquito vector, and the NCI60 human cancer cell line panel of 60 human tumor cell lines. Toxicological, pharmacokinetic and metabolic properties were collected on all the compounds, assisting in the selection of the most promising candidates for murine proof-of-concept experiments and medicinal chemistry programs. The data for all of these assays are presented and analyzed to show how outstanding leads for many indications can be selected. These results reveal the immense potential for translating the dispersed expertise in biological assays involving human pathogens into drug discovery starting points, by providing open access to new families of molecules, and emphasize how a small additional investment made to help acquire and distribute compounds, and sharing the data, can catalyze drug discovery for dozens of different indications. Another lesson is that when multiple screens from different groups are run on the same library, results can be integrated quickly to select the most valuable starting points for subsequent medicinal chemistry efforts.

"From safe source to safe sink" development of colorimetric assay for gabapentin in bulk drug and capsules using naturally derived genipin.

PubMed

Winotapun, Weerapath; Kongpakwattana, Khachen; Dejpittayanunt, Sirirat; Pathomcharoensukchai, Suwaparp; Suksaran, Udomluck; Nuntharatanapong, Nopparat; Rojanarata, Theerasak

2012-09-15

A novel colorimetric assay for gabapentin in bulk drug and capsules has been developed via a safety-and-sustainability concerning concept. The method relied on the reaction of primary amino group of drug with non-toxic and eco-friendly genipin in totally aqueous medium to form the blue product which was subsequently measured by visible spectrophotometry at 590 nm. Under the optimized conditions, Beer's law was obeyed in the concentration range of 0.15-0.50 mM (r(2)=0.9998). It was accurate, precise and insensitive to the interferences from all related compounds specified in the United States Pharmacopeia as well as commonly used excipients. Furthermore, it gave the assay results in agreement with the pharmacopeial chromatographic method. Owing to the environmental concern and responsibility, a fast and facile method was also proposed for the treatment of waste generated from the assay based on the decoloration by using gypsum as a cheap and commonly available adsorbent. After the treatment, more than 95% of the initial blue product was removed from the waste solution and the treated waste was proven to be safe for aquatic organisms, as studied in brine shrimp and guppy fishes. Therefore, this work not only reports for the first time the application of naturally derived genipin to drug analysis, but also presents a new and contemporary paradigm that illustrates the fully benign-by-design development of the analytical methodologies in the era of Green Chemistry, starting from the safe source of reagents toward the safe sink when waste is released into the environment. Copyright © 2012 Elsevier B.V. All rights reserved.

Enhanced styrene recovery from waste polystyrene pyrolysis using response surface methodology coupled with Box-Behnken design.

PubMed

Mo, Yu; Zhao, Lei; Wang, Zhonghui; Chen, Chia-Lung; Tan, Giin-Yu Amy; Wang, Jing-Yuan

2014-04-01

A work applied response surface methodology coupled with Box-Behnken design (RSM-BBD) has been developed to enhance styrene recovery from waste polystyrene (WPS) through pyrolysis. The relationship between styrene yield and three selected operating parameters (i.e., temperature, heating rate, and carrier gas flow rate) was investigated. A second order polynomial equation was successfully built to describe the process and predict styrene yield under the study conditions. The factors identified as statistically significant to styrene production were: temperature, with a quadratic effect; heating rate, with a linear effect; carrier gas flow rate, with a quadratic effect; interaction between temperature and carrier gas flow rate; and interaction between heating rate and carrier gas flow rate. The optimum conditions for the current system were determined to be at a temperature range of 470-505°C, a heating rate of 40°C/min, and a carrier gas flow rate range of 115-140mL/min. Under such conditions, 64.52% WPS was recovered as styrene, which was 12% more than the highest reported yield for reactors of similar size. It is concluded that RSM-BBD is an effective approach for yield optimization of styrene recovery from WPS pyrolysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

Quantitative and Cost Comparison of Ultrasensitive Human Immunodeficiency Virus Type 1 RNA Viral Load Assays: Bayer bDNA Quantiplex Versions 3.0 and 2.0 and Roche PCR Amplicor Monitor Version 1.5

PubMed Central

Elbeik, Tarek; Charlebois, Edwin; Nassos, Patricia; Kahn, James; Hecht, Frederick M.; Yajko, David; Ng, Valerie; Hadley, Keith

2000-01-01

Quantification of human immunodeficiency virus type 1 (HIV-1) RNA as a measure of viral load has greatly improved the monitoring of therapies for infected individuals. With the significant reductions in viral load now observed in individuals treated with highly active anti-retroviral therapy (HAART), viral load assays have been adapted to achieve greater sensitivity. Two commercially available ultrasensitive assays, the Bayer Quantiplex HIV-1 bDNA version 3.0 (bDNA 3.0) assay and the Roche Amplicor HIV-1 Monitor Ultrasensitive version 1.5 (Amplicor 1.5) assay, are now being used to monitor HIV-1-infected individuals. Both of these ultrasensitive assays have a reported lower limit of 50 HIV-1 RNA copies/ml and were developed from corresponding older generation assays with lower limits of 400 to 500 copies/ml. However, the comparability of viral load data generated by these ultrasensitive assays and the relative costs of labor, disposables, and biohazardous wastes were not determined in most cases. In this study, we used matched clinical plasma samples to compare the quantification of the newer bDNA 3.0 assay with that of the older bDNA 2.0 assay and to compare the quantification and costs of the bDNA 3.0 assay and the Amplicor 1.5 assay. We found that quantification by the bDNA 3.0 assay was approximately twofold higher than that by the bDNA 2.0 assay and was highly correlated to that by the Amplicor 1.5 assay. Moreover, cost analysis based on labor, disposables, and biohazardous wastes showed significant savings with the bDNA 3.0 assay as compared to the costs of the Amplicor 1.5 assay. PMID:10699005

Quantitative and cost comparison of ultrasensitive human immunodeficiency virus type 1 RNA viral load assays: Bayer bDNA quantiplex versions 3.0 and 2.0 and Roche PCR Amplicor monitor version 1.5.

PubMed

Elbeik, T; Charlebois, E; Nassos, P; Kahn, J; Hecht, F M; Yajko, D; Ng, V; Hadley, K

2000-03-01

Quantification of human immunodeficiency virus type 1 (HIV-1) RNA as a measure of viral load has greatly improved the monitoring of therapies for infected individuals. With the significant reductions in viral load now observed in individuals treated with highly active anti-retroviral therapy (HAART), viral load assays have been adapted to achieve greater sensitivity. Two commercially available ultrasensitive assays, the Bayer Quantiplex HIV-1 bDNA version 3.0 (bDNA 3.0) assay and the Roche Amplicor HIV-1 Monitor Ultrasensitive version 1.5 (Amplicor 1.5) assay, are now being used to monitor HIV-1-infected individuals. Both of these ultrasensitive assays have a reported lower limit of 50 HIV-1 RNA copies/ml and were developed from corresponding older generation assays with lower limits of 400 to 500 copies/ml. However, the comparability of viral load data generated by these ultrasensitive assays and the relative costs of labor, disposables, and biohazardous wastes were not determined in most cases. In this study, we used matched clinical plasma samples to compare the quantification of the newer bDNA 3.0 assay with that of the older bDNA 2.0 assay and to compare the quantification and costs of the bDNA 3.0 assay and the Amplicor 1.5 assay. We found that quantification by the bDNA 3.0 assay was approximately twofold higher than that by the bDNA 2.0 assay and was highly correlated to that by the Amplicor 1.5 assay. Moreover, cost analysis based on labor, disposables, and biohazardous wastes showed significant savings with the bDNA 3.0 assay as compared to the costs of the Amplicor 1.5 assay.

SCALE UP OF CERAMIC WASTE FORMS FOR THE EBR-II SPENT FUEL TREATMENT PROCESS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Matthew C. Morrison; Kenneth J. Bateman; Michael F. Simpson

2010-11-01

ABSTRACT SCALE UP OF CERAMIC WASTE FORMS FOR THE EBR-II SPENT FUEL TREATMENT PROCESS Matthew C. Morrison, Kenneth J. Bateman, Michael F. Simpson Idaho National Laboratory, P.O. Box 1625, Idaho Falls, ID 83415 The ceramic waste process is the intended method for disposing of waste salt electrolyte, which contains fission products from the fuel-processing electrorefiners (ER) at the INL. When mixed and processed with other materials, the waste salt can be stored in a durable ceramic waste form (CWF). The development of the CWF has recently progressed from small-scale testing and characterization to full-scale implementation and experimentation using surrogate materialsmore » in lieu of the ER electrolyte. Two full-scale (378 kg and 383 kg) CWF test runs have been successfully completed with final densities of 2.2 g/cm3 and 2.1 g/cm3, respectively. The purpose of the first CWF was to establish material preparation parameters. The emphasis of the second pre-qualification test run was to evaluate a preliminary multi-section CWF container design. Other considerations were to finalize material preparation parameters, measure the material height as it consolidates in the furnace, and identify when cracking occurs during the CWF cooldown process.« less

A COMPREHENSIVE TECHNICAL REVIEW OF THE DEMONSTRATION BULK VITRIFICATION SYSTEM

DOE Office of Scientific and Technical Information (OSTI.GOV)

SCHAUS, P.S.

2006-09-29

In May 2006, CH2M Hill Hanford Group, Inc. chartered an Expert Review Panel (ERP) to review the current status of the Demonstration Bulk Vitrification System (DBVS). It is the consensus of the ERP that bulk vitrification is a technology that requires further development and evaluation to determine its potential for meeting the Hanford waste stabilization mission. No fatal flaws (issues that would jeopardize the overall DBVS mission that cannot be mitigated) were found, given the current state of the project. However, a number of technical issues were found that could significantly affect the project's ability to meet its overall missionmore » as stated in the project ''Justification of Mission Need'' document, if not satisfactorily resolved. The ERP recognizes that the project has changed from an accelerated schedule demonstration project to a formally chartered project that must be in full compliance with DOE 413.3 requirements. The perspective of the ERP presented herein, is measured against the formally chartered project as stated in the approved Justification of Mission Need document. A justification of Mission Need document was approved in July 2006 which defined the objectives for the DBVS Project. In this document, DOE concluded that bulk vitrification is a viable technology that requires additional development to determine its potential applicability to treatment of a portion of the Hanford low activity waste. The DBVS mission need statement now includes the following primary objectives: (1) process approximately 190,000 gallons of Tank S-109 waste into fifty 100 metric ton boxes of vitrified product; (2) store and dispose of these boxes at Hanford's Integrated Disposal Facility (IDF); (3) evaluate the waste form characteristics; (4) gather pilot plant operability data, and (5) develop the overall life cycle system performance of bulk vitrification and produce a comparison of the bulk vitrification process to building a second LAW Immobilization facility or other supplemental treatment alternatives as provided in M-62-08.« less

Molecular Cloning of a cDNA Encoding for Taenia solium TATA-Box Binding Protein 1 (TsTBP1) and Study of Its Interactions with the TATA-Box of Actin 5 and Typical 2-Cys Peroxiredoxin Genes.

PubMed

Rodríguez-Lima, Oscar; García-Gutierrez, Ponciano; Jiménez, Lucía; Zarain-Herzberg, Ángel; Lazzarini, Roberto; Landa, Abraham

2015-01-01

TATA-box binding protein (TBP) is an essential regulatory transcription factor for the TATA-box and TATA-box-less gene promoters. We report the cloning and characterization of a full-length cDNA that encodes a Taenia solium TATA-box binding protein 1 (TsTBP1). Deduced amino acid composition from its nucleotide sequence revealed that encodes a protein of 238 residues with a predicted molecular weight of 26.7 kDa, and a theoretical pI of 10.6. The NH2-terminal domain shows no conservation when compared with to pig and human TBP1s. However, it shows high conservation in size and amino acid identity with taeniids TBP1s. In contrast, the TsTBP1 COOH-terminal domain is highly conserved among organisms, and contains the amino acids involved in interactions with the TATA-box, as well as with TFIIA and TFIIB. In silico TsTBP1 modeling reveals that the COOH-terminal domain forms the classical saddle structure of the TBP family, with one α-helix at the end, not present in pig and human. Native TsTBP1 was detected in T. solium cysticerci´s nuclear extract by western blot using rabbit antibodies generated against two synthetic peptides located in the NH2 and COOH-terminal domains of TsTBP1. These antibodies, through immunofluorescence technique, identified the TBP1 in the nucleus of cells that form the bladder wall of cysticerci of Taenia crassiceps, an organism close related to T. solium. Electrophoretic mobility shift assays using nuclear extracts from T. solium cysticerci and antibodies against the NH2-terminal domain of TsTBP1 showed the interaction of native TsTBP1 with the TATA-box present in T. solium actin 5 (pAT5) and 2-Cys peroxiredoxin (Ts2-CysPrx) gene promoters; in contrast, when antibodies against the anti-COOH-terminal domain of TsTBP1 were used, they inhibited the binding of TsTBP1 to the TATA-box of the pAT5 promoter gene.

Apparatus and method for quantitative assay of samples of transuranic waste contained in barrels in the presence of matrix material

DOEpatents

Caldwell, J.T.; Herrera, G.C.; Hastings, R.D.; Shunk, E.R.; Kunz, W.E.

1987-08-28

Apparatus and method for performing corrections for matrix material effects on the neutron measurements generated from analysis of transuranic waste drums using the differential-dieaway technique. By measuring the absorption index and the moderator index for a particular drum, correction factors can be determined for the effects of matrix materials on the ''observed'' quantity of fissile and fertile material present therein in order to determine the actual assays thereof. A barrel flux monitor is introduced into the measurement chamber to accomplish these measurements as a new contribution to the differential-dieaway technology. 9 figs.

In Vitro Assays for Assessment of Androgenic and Estrogenic Activity of Defined Mixtures and Complex Environmental Samples

EPA Science Inventory

Point sources of endocrine active compounds to aquatic environments such as waste water treatment plants, pulp and paper mills, and animal feeding operations invariably contain complex mixtures of chemicals. The current study investigates the use of targeted in vitro assays des...

Acute toxicity assessment of Polish (waste) water with a microplate-based Hydra attenuata assay: a comparison with the Microtox test.

PubMed

Pardos, M; Benninghoff, C; Guéguen, C; Thomas, R; Dobrowolski, J; Dominik, J

1999-12-15

The use of Hydra attenuata in acute toxicity assessment is a potentially useful tool in (waste) water biomonitoring. The purpose of this study was to compare the sensitivity of H. attenuata with the extensively used Microtox test on 14 (waste) water samples from the Kraków region (South Poland). To this end, specific morphological changes displayed by the freshwater cnidarian Hydra attenuata (lethal LC50s and sublethal EC50s effects) and bioluminescence of the marine bacteria Vibrio fisheri (Microtox) were compared. Clearly, the Hydra assay was the more sensitive indicator of toxicity. No relationship was found among Hydra toxicological responses and water levels of As, Cd, Co, Cu, Pb and Zn. However, it appeared that toxicity to Hydra might be due to ammonia levels. Additional studies to better circumscribe the tolerance of H. attenuata to 'natural' water characteristics are needed.

The pregnane X receptor down‐regulates organic cation transporter 1 (SLC22A1) in human hepatocytes by competing for (“squelching”) SRC‐1 coactivator

PubMed Central

Hyrsova, Lucie; Smutny, Tomas; Carazo, Alejandro; Moravcik, Stefan; Mandikova, Jana; Trejtnar, Frantisek; Gerbal‐Chaloin, Sabine

2016-01-01

Background and Purpose The organic cation transporter 1 (OCT1) transports cationic drugs into hepatocytes. The high hepatic expression of OCT1 is controlled by the HNF4α and USF transcription factors. Pregnane X receptor (PXR) mediates induction of the principal xenobiotic metabolizing enzymes and transporters in the liver. Here, we have assessed the down‐regulation of OCT1 expression by PXR activation. Experimental Approach We used primary human hepatocytes and related cell lines to measure OCT1 expression and activity, by assaying MPP+ accumulation. Western blotting, qRT‐PCR, the OCT1 promoter gene reporter constructs and chromatin immunoprecipitation assays were also used. Key Results OCT1 mRNA in human hepatocytes was down‐regulated along with reduced [3H]MPP+ accumulation in differentiated HepaRG cells after treatment with rifampicin. Rifampicin and hyperforin as well as the constitutively active PXR mutant T248D suppressed activity of the 1.8 kb OCT1 promoter construct in gene reporter assays. Silencing of both PXR and HNF4α in HepaRG cells blocked the PXR ligand‐mediated down‐regulation of OCT1 expression. The mutation of HNF4α and USF1 (E‐box) responsive elements reversed the PXR‐mediated inhibition in gene reporter assays. Chromatin immunoprecipitation assays indicated that PXR activation sequestrates the SRC‐1 coactivator from the HNF4α response element and E‐box of the OCT1 promoter. Consistent with these findings, exogenous overexpression of the SRC‐1, but not the PGC1α coactivator, relieved the PXR‐mediated repression of OCT1 transactivation. Conclusions and Implications PXR ligands reduced the HNF4α‐mediated and USF‐mediated transactivation of OCT1 gene expression by competing for SRC‐1 and decreased delivery of a model OCT1 substrate into hepatocytes. PMID:26920453

Non-radioactive TRF assay modifications to improve telomeric DNA detection efficiency in plants

PubMed Central

Nigmatullina, Liliia R.; Sharipova, Margarita R.; Shakirov, Eugene V.

2016-01-01

The length of telomeric DNA is often considered a cellular biomarker of aging and general health status. Several telomere length measuring assays have been developed, of which the most common is the Telomere Restriction Fragment (TRF) analysis, which typically involves the use of radioactively labeled oligonucleotide probes. While highly effective, this method potentially poses substantial health concerns and generates radioactive waste. Digoxigenin (DIG) alternatives to radioactive probes have been developed and used successfully in a number of assays. Here we optimize the DIG protocol to measure telomere length in the model plant Arabidopsis thaliana and present evidence that this approach can be used successfully to efficiently and accurately measure telomere length in plants. Specifically, hybridization temperature of 42 °C instead of the typical 55 °C appears to generate stronger signals. In addition, DIG incorporation at 5′-end instead of 3′-end of the labeled oligonucleotide greatly enhances signal. We conclude that non-radioactive TRF assays can be as efficient as radioactive methods in detecting and measuring telomere length in plants, making this assay suitable for medical and research laboratories unable to utilize radioactivity due to hazardous waste disposal and safety concerns. PMID:28133587

Evaluation of antioxidant potential of Artocarpus heterophyllus L. J33 variety fruit waste from different extraction methods and identification of phenolic constituents by LCMS.

PubMed

Daud, Mohd Nazrul Hisham; Fatanah, Dian Nashiela; Abdullah, Noriham; Ahmad, Rohaya

2017-10-01

Artocarpus heterophyllus J33 (AhJ33) fruit is a popular and valuable jackfruit variety in Malaysia. For export, the pulp has to be separated from the skin which is usually discarded. Hence, the conversion of the fruit waste to food products with economic value needs to be explored utilizing the waste to wealth concept. This paper reports the evaluation of antioxidant potential of AhJ33 fruit waste (rind and rachis) extracts from three different extraction methods (maceration, percolation and Soxhlet). The antioxidant potential was assessed by DPPH radical scavenging, FRAP and β-carotene bleaching assays. The total phenolic and total flavonoid contents were estimated by TPC and the TFC assays. For both rind and rachis, the maceration technique yielded extracts with the strongest antioxidant activities which correlated with the highest TPC and TFC values. TOF LCMS analyses identified two phenolic acids as the major constituents responsible for the antioxidant activity of the active extracts. Copyright © 2017 Elsevier Ltd. All rights reserved.

S-adenosylmethionine directly inhibits binding of 30S ribosomal subunits to the SMK box translational riboswitch RNA

PubMed Central

Fuchs, Ryan T.; Grundy, Frank J.; Henkin, Tina M.

2007-01-01

The SMK box is a conserved riboswitch motif found in the 5′ untranslated region of metK genes [encoding S-adenosylmethionine (SAM) synthetase] in lactic acid bacteria, including Enterococcus, Streptococcus, and Lactococcus sp. Previous studies showed that this RNA element binds SAM in vitro, and SAM binding causes a structural rearrangement that sequesters the Shine–Dalgarno (SD) sequence by pairing with an anti-SD (ASD) element. A model was proposed in which SAM binding inhibits metK translation by preventing binding of the ribosome to the SD region of the mRNA. In the current work, the addition of SAM was shown to inhibit binding of 30S ribosomal subunits to SMK box RNA; in contrast, the addition of S-adenosylhomocysteine (SAH) had no effect. A mutant RNA, which has a disrupted SD-ASD pairing, was defective in SAM binding and showed no reduction of ribosome binding in the presence of SAM, whereas a compensatory mutation that restored SD-ASD pairing restored the response to SAM. Primer extension inhibition assays provided further evidence for SD-ASD pairing in the presence of SAM. These results strongly support the model that SMK box translational repression operates through occlusion of the ribosome binding site and that SAM binding requires the SD-ASD pairing. PMID:17360376

The SAM-responsive SMK box is a reversible riboswitch

PubMed Central

Smith, Angela M.; Fuchs, Ryan T.; Grundy, Frank J.; Henkin, Tina M.

2010-01-01

The SMK (SAM-III) box is an S-adenosylmethionine (SAM)-responsive riboswitch found in the 5′ untranslated region of metK genes, encoding SAM synthetase, in many members of the Lactobacillales. SAM binding causes a structural rearrangement in the RNA that sequesters the Shine-Dalgarno (SD) sequence by pairing with a complementary anti-SD (ASD) sequence; sequestration of the SD sequence inhibits binding of the 30S ribosomal subunit and prevents translation initiation. We observed a slight increase in the half-life of the metK transcript in vivo when Enterococcus faecalis cells were depleted for SAM, but no significant change in overall transcript abundance, consistent with the model that this riboswitch regulates at the level of translation initiation. The half-life of the SAM-SMK box RNA complex in vitro is shorter than that of the metK transcript in vivo, raising the possibility of reversible binding of SAM. We used a fluorescence assay to directly visualize reversible switching between the SAM-free and SAM-bound conformations. We propose that the SMK box riboswitch can make multiple SAM-dependent regulatory decisions during the lifetime of the transcript in vivo, acting as a reversible switch that allows the cell to respond rapidly to fluctuations in SAM pools by modulating expression of the SAM synthetase gene. PMID:21143313

In vitro mapping of Myotonic Dystrophy (DM) gene promoter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Storbeck, C.J.; Sabourin, L.; Baird, S.

1994-09-01

The Myotonic Dystrophy Kinase (DMK) gene has been cloned and shared homology to serine/threonine protein kinases. Overexpression of this gene in stably transfected mouse myoblasts has been shown to inhibit fusion into myotubes while myoblasts stably transfected with an antisense construct show increased fusion potential. These experiments, along with data showing that the DM gene is highly expressed in muscle have highlighted the possibility of DMK being involved in myogenesis. The promoter region of the DM gene lacks a consensus TATA box and CAAT box, but harbours numerous transcription binding sites. Clones containing extended 5{prime} upstream sequences (UPS) of DMKmore » only weakly drive the reporter gene chloramphenicol acetyl transferase (CAT) when transfected into C2C12 mouse myoblasts. However, four E-boxes are present in the first intron of the DM gene and transient assays show increased expression of the CAT gene when the first intron is present downstream of these 5{prime} UPS in an orientation dependent manner. Comparison between mouse and human sequence reveals that the regions in the first intron where the E-boxes are located are highly conserved. The mapping of the promoter and the importance of the first intron in the control of DMK expression will be presented.« less

Transcriptional Regulation of Fruit Ripening by Tomato FRUITFULL Homologs and Associated MADS Box Proteins[W

PubMed Central

Fujisawa, Masaki; Shima, Yoko; Nakagawa, Hiroyuki; Kitagawa, Mamiko; Kimbara, Junji; Nakano, Toshitsugu; Kasumi, Takafumi; Ito, Yasuhiro

2014-01-01

The tomato (Solanum lycopersicum) MADS box FRUITFULL homologs FUL1 and FUL2 act as key ripening regulators and interact with the master regulator MADS box protein RIPENING INHIBITOR (RIN). Here, we report the large-scale identification of direct targets of FUL1 and FUL2 by transcriptome analysis of FUL1/FUL2 suppressed fruits and chromatin immunoprecipitation coupled with microarray analysis (ChIP-chip) targeting tomato gene promoters. The ChIP-chip and transcriptome analysis identified FUL1/FUL2 target genes that contain at least one genomic region bound by FUL1 or FUL2 (regions that occur mainly in their promoters) and exhibit FUL1/FUL2-dependent expression during ripening. These analyses identified 860 direct FUL1 targets and 878 direct FUL2 targets; this set of genes includes both direct targets of RIN and nontargets of RIN. Functional classification of the FUL1/FUL2 targets revealed that these FUL homologs function in many biological processes via the regulation of ripening-related gene expression, both in cooperation with and independent of RIN. Our in vitro assay showed that the FUL homologs, RIN, and tomato AGAMOUS-LIKE1 form DNA binding complexes, suggesting that tetramer complexes of these MADS box proteins are mainly responsible for the regulation of ripening. PMID:24415769

Cold shock protein YB-1 is involved in hypoxia-dependent gene transcription.

PubMed

Rauen, Thomas; Frye, Bjoern C; Wang, Jialin; Raffetseder, Ute; Alidousty, Christina; En-Nia, Abdelaziz; Floege, Jürgen; Mertens, Peter R

2016-09-16

Hypoxia-dependent gene regulation is largely orchestrated by hypoxia-inducible factors (HIFs), which associate with defined nucleotide sequences of hypoxia-responsive elements (HREs). Comparison of the regulatory HRE within the 3' enhancer of the human erythropoietin (EPO) gene with known binding motifs for cold shock protein Y-box (YB) protein-1 yielded strong similarities within the Y-box element and 3' adjacent sequences. DNA binding assays confirmed YB-1 binding to both, single- and double-stranded HRE templates. Under hypoxia, we observed nuclear shuttling of YB-1 and co-immunoprecipitation assays demonstrated that YB-1 and HIF-1α physically interact with each other. Cellular YB-1 depletion using siRNA significantly induced hypoxia-dependent EPO production at both, promoter and mRNA level. Vice versa, overexpressed YB-1 significantly reduced EPO-HRE-dependent gene transcription, whereas this effect was minor under normoxia. HIF-1α overexpression induced hypoxia-dependent gene transcription through the same element and accordingly, co-expression with YB-1 reduced HIF-1α-mediated EPO induction under hypoxic conditions. Taken together, we identified YB-1 as a novel binding factor for HREs that participates in fine-tuning of the hypoxia transcriptome. Copyright © 2016 Elsevier Inc. All rights reserved.

Human Nup98 regulates the localization and activity of DExH/D-box helicase DHX9

PubMed Central

Capitanio, Juliana S; Montpetit, Ben; Wozniak, Richard W

2017-01-01

Beyond their role at nuclear pore complexes, some nucleoporins function in the nucleoplasm. One such nucleoporin, Nup98, binds chromatin and regulates gene expression. To gain insight into how Nup98 contributes to this process, we focused on identifying novel binding partners and understanding the significance of these interactions. Here we report on the identification of the DExH/D-box helicase DHX9 as an intranuclear Nup98 binding partner. Various results, including in vitro assays, show that the FG/GLFG region of Nup98 binds to N- and C-terminal regions of DHX9 in an RNA facilitated manner. Importantly, binding of Nup98 stimulates the ATPase activity of DHX9, and a transcriptional reporter assay suggests Nup98 supports DHX9-stimulated transcription. Consistent with these observations, our analysis revealed that Nup98 and DHX9 bind interdependently to similar gene loci and their transcripts. Based on our results, we propose that Nup98 functions as a co-factor that regulates DHX9 and, potentially, other RNA helicases. DOI: http://dx.doi.org/10.7554/eLife.18825.001 PMID:28221134

UV and arsenate toxicity: a specific and sensitive yeast bioluminescence assay.

PubMed

Bakhrat, Anya; Eltzov, Evgeni; Finkelstein, Yishay; Marks, Robert S; Raveh, Dina

2011-06-01

We describe a Saccharomyces cerevisiae bioluminescence assay for UV and arsenate in which bacterial luciferase genes are regulated by the promoter of the yeast gene, UFO1. UFO1 encodes the F-box subunit of the Skp1–Cdc53–F-box protein ubiquitin ligase complex and is induced by DNA damage and by arsenate. We engineered the UFO1 promoter into an existing yeast bioreporter that employs human genes for detection of steroid hormone-disrupting compounds in water bodies. Our analysis indicates that use of an endogenous yeast promoter in different mutant backgrounds allows discrimination between different environmental signals. The UFO1-engineered yeast give a robust bioluminescence response to UVB and can be used for evaluating UV protective sunscreens. They are also effective in detecting extremely low concentrations of arsenate, particularly in pdr5Δ mutants that lack a mechanism to extrude toxic chemicals; however, they do not respond to cadmium or mercury. Combined use of endogenous yeast promoter elements and mutants of stress response pathways may facilitate development of high-specificity yeast bioreporters able to discriminate between closely related chemicals present together in the environment.

Using ATP-driven bioluminescence assay to monitor microbial safety in a contemporary human cadaver laboratory.

PubMed

Benninger, Brion; Maier, Thomas

2015-03-01

The objective of this study was to utilize a cost-effective method for assessing the levels of bacterial, yeast, and mold activity during a human dissection laboratory course. Nowadays, compliance with safety regulations is policed by institutions at higher standards than ever before. Fear of acquiring an unknown infection is one of the top concerns of professional healthcare students, and it provokes anti-laboratory anxiety. Human cadavers are not routinely tested for bacteria and viruses prior to embalming. Human anatomy dissecting rooms that house embalmed cadavers are normally cleaned after the dissected cadavers have been removed. There is no evidence that investigators have ever assessed bacterial and fungal activities using adenosine triphosphate (ATP)-driven bioluminescence assays. A literature search was conducted on texts, journals, and websites regarding bacterial, yeast, and mold activities in an active cadaver laboratory. Midway into a clinical anatomy course, ATP bioluminescence assays were used to swab various sites within the dissection room, including entrance and exiting door handles, water taps, cadaver tables, counter tops, imaging material, X-ray box switches, and the cadaver surfaces. The results demonstrated very low activities on cadaver tables, washing up areas, and exiting door handles. There was low activity on counter tops and X-ray boxes. There was medium activity on the entrance door handles. These findings suggest an inexpensive and accurate method for monitoring safety compliance and microbial activity. Students can feel confident and safe in the environment in which they work. © 2014 Wiley Periodicals, Inc.

ABCB1 regulation through LRPPRC is influenced by the methylation status of the GC -100 box in its promoter

PubMed Central

Corrêa, Stephany; Binato, Renata; Du Rocher, Bárbara; Ferreira, Gerson; Cappelletti, Paola; Soares-Lima, Sheila; Pinto, Luis Felipe; Mencalha, André; Abdelhay, Eliana

2014-01-01

One of the potential mechanisms of imatinib mesylate (IM) resistance in chronic myeloid leukemia (CML) is increased level of P-glycoprotein (Pgp). Pgp is an efflux pump capable of activating the multidrug resistance (MDR) phenotype. The gene encoding Pgp (ABCB1) has several binding sites in its promoter region, along with CpG islands and GC boxes, involved in its epigenetic control. In previous work, we performed a proteomic study to identify proteins involved in IM cross-resistance in acute leukemia. Among these proteins, we identified LRPPRC as a potential regulator of ABCB1 transcription via an invMED1 binding site in ABCB1. Interestingly, this invMED1 binding site overlaps with the GC -100 box. In this work, we investigated the potential role of LRPPRC in the regulation of ABCB1 transcriptional activity in CML resistance. In addition, we evaluated the potential connection between this regulation and the methylation status of the ABCB1 promoter in its GC -100 box. Our results show that LRPPRC binds prominently to the ABCB1 promoter in Lucena cells, an IM-resistant cell line. Luciferase assays showed that ABCB1 transcription is positively regulated by LRPPRC upon its knockdown. Pyrosequencing analysis showed that the ABCB1 promoter is differentially methylated at its GC -100 box in K562 cells compared with Lucena cells, and in CML patients with different response to IM. Chromatin immunoprecipitation and Pgp expression after DNA demethylation treatment showed that LRPPRC binding is affected by the methylation status of ABCB1 GC -100 box. Taken together, our findings indicate that LRPPRC is a transcription factor related to ABCB1 expression and highlight the importance of epigenetic regulation in CML resistance. PMID:25089713

ABCB1 regulation through LRPPRC is influenced by the methylation status of the GC -100 box in its promoter.

PubMed

Corrêa, Stephany; Binato, Renata; Du Rocher, Bárbara; Ferreira, Gerson; Cappelletti, Paola; Soares-Lima, Sheila; Pinto, Luis Felipe; Mencalha, André; Abdelhay, Eliana

2014-08-01

One of the potential mechanisms of imatinib mesylate (IM) resistance in chronic myeloid leukemia (CML) is increased level of P-glycoprotein (Pgp). Pgp is an efflux pump capable of activating the multidrug resistance (MDR) phenotype. The gene encoding Pgp (ABCB1) has several binding sites in its promoter region, along with CpG islands and GC boxes, involved in its epigenetic control. In previous work, we performed a proteomic study to identify proteins involved in IM cross-resistance in acute leukemia. Among these proteins, we identified LRPPRC as a potential regulator of ABCB1 transcription via an invMED1 binding site in ABCB1. Interestingly, this invMED1 binding site overlaps with the GC -100 box. In this work, we investigated the potential role of LRPPRC in the regulation of ABCB1 transcriptional activity in CML resistance. In addition, we evaluated the potential connection between this regulation and the methylation status of the ABCB1 promoter in its GC -100 box. Our results show that LRPPRC binds prominently to the ABCB1 promoter in Lucena cells, an IM-resistant cell line. Luciferase assays showed that ABCB1 transcription is positively regulated by LRPPRC upon its knockdown. Pyrosequencing analysis showed that the ABCB1 promoter is differentially methylated at its GC -100 box in K562 cells compared with Lucena cells, and in CML patients with different response to IM. Chromatin immunoprecipitation and Pgp expression after DNA demethylation treatment showed that LRPPRC binding is affected by the methylation status of ABCB1 GC -100 box. Taken together, our findings indicate that LRPPRC is a transcription factor related to ABCB1 expression and highlight the importance of epigenetic regulation in CML resistance.

Molecular cloning and characterization of SoxB2 gene from Zhikong scallop Chlamys farreri

NASA Astrophysics Data System (ADS)

He, Yan; Bao, Zhenmin; Guo, Huihui; Zhang, Yueyue; Zhang, Lingling; Wang, Shi; Hu, Jingjie; Hu, Xiaoli

2013-11-01

The Sox proteins play critical roles during the development of animals, including sex determination and central nervous system development. In this study, the SoxB2 gene was cloned from a mollusk, the Zhikong scallop ( Chlamys farreri), and characterized with respect to phylogeny and tissue distribution. The full-length cDNA and genomic DNA sequences of C. farreri SoxB2 ( Cf SoxB2) were obtained by rapid amplification of cDNA ends and genome walking, respectively, using a partial cDNA fragment from the highly conserved DNA-binding domain, i.e., the High Mobility Group (HMG) box. The full-length cDNA sequence of Cf SoxB2 was 2 048 bp and encoded 268 amino acids protein. The genomic sequence was 5 551 bp in length with only one exon. Several conserved elements, such as the TATA-box, GC-box, CAAT-box, GATA-box, and Sox/sry-sex/testis-determining and related HMG box factors, were found in the promoter region. Furthermore, real-time quantitative reverse transcription PCR assays were carried out to assess the mRNA expression of Cf SoxB 2 in different tissues. SoxB2 was highly expressed in the mantle, moderately in the digestive gland and gill, and weakly expressed in the gonad, kidney and adductor muscle. In male and female gonads at different developmental stages of reproduction, the expression levels of Cf SoxB2 were similar. Considering the specific expression and roles of SoxB 2 in other animals, in particular vertebrates, and the fact that there are many pallial nerves in the mantle, cerebral ganglia in the digestive gland and gill nerves in gill, we propose a possible essential role in nervous tissue function for Sox B 2 in C. farreri.

Optimization and preliminary characterization of venom isolated from 3 medically important jellyfish: the box (Chironex fleckeri), Irukandji (Carukia barnesi), and blubber (Catostylus mosaicus) jellyfish.

PubMed

Wiltshire, C J; Sutherland, S K; Fenner, P J; Young, A R

2000-01-01

To optimize venom extraction and to undertake preliminary biochemical studies of venom from the box jellyfish (Chironex fleckeri), the Irukandji jellyfish (Carukia barnesi), and the blubber jellyfish (Catostylus mosaicus). Lyophilized crude venoms from box jellyfish tentacles and whole Irukandji jellyfish were prepared in water by homogenization, sonication, and rapid freeze thawing. A second technique, consisting of grinding samples with a glass mortar and pestle and using phosphate-buffered saline, was used to prepare crude venom from isolated nematocysts of the box jellyfish, the bells of Irukandji jellyfish, and the oral lobes of blubber jellyfish. Venoms were compared by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot test. Toxicity of some venoms was determined by intravenous median lethal dose assay in mice. Different venom extraction techniques produced significantly different crude venoms for both box and Irukandji jellyfish. Irukandji and blubber venom SDS-PAGE protein profiles were established for the first time. Analysis of Western blot tests revealed that box jellyfish antivenin reacted specifically with the venom of each jellyfish. Toxicity was found in Irukandji jellyfish venom derived by use of the mortar-and-pestle method, but not in the lyophilized venom. Glass mortar-and-pestle grinding and use of an appropriate buffer was found to be a simple and suitable method for the preparation of venom from each jellyfish species studied. This study contributes to biochemical investigations of jellyfish venoms, particularly the venom of the Irukandji jellyfish, for which there are, to our knowledge, no published studies. It also highlights the importance of optimizing venom extraction as the first step toward understanding the complex biological effects of jellyfish venoms.

Increased actin polymerization reduces the inhibition of serum response factor activity by Yin Yang 1.

PubMed Central

Ellis, Peter D; Martin, Karen M; Rickman, Colin; Metcalfe, James C; Kemp, Paul R

2002-01-01

Recent evidence has implicated CC(A/T(richG))GG (CArG) boxes, binding sites for serum response factor (SRF), in the regulation of expression of a number of genes in response to changes in the actin cytoskeleton. In many cases, the activity of SRF at CArG boxes is modulated by transcription factors binding to overlapping (e.g. Yin Yang 1, YY1) or adjacent (e.g. ets) binding sites. However, the mechanisms by which SRF activity is regulated by the cytoskeleton have not been determined. To investigate these mechanisms, we screened for cells that did or did not increase the activity of a fragment of the promoter for a smooth-muscle (SM)-specific gene SM22alpha, in response to changes in actin cytoskeletal polymerization induced by LIM kinase. These experiments showed that vascular SM cells (VSMCs) and C2C12 cells increased the activity of promoters containing at least one of the SM22alpha CArG boxes (CArG near) in response to LIM kinase, whereas P19 cells did not. Bandshift assays using a probe to CArG near showed that P19 cells lacked detectable YY1 DNA binding to the CArG box in contrast with the other two cell types. Expression of YY1 in P19 cells inhibited SM22alpha promoter activity and conferred responsiveness to LIM kinase. Mutation of the CArG box to inhibit YY1 or SRF binding indicated that both factors were required for the LIM kinase response in VSMCs and C2C12 cells. The data indicate that changes in the actin cytoskeletal organization modify SRF activity at CArG boxes by modulating YY1-dependent inhibition. PMID:12023898

Biodegradation of waste lubricants by a newly isolated Ochrobactrum sp. C1.

PubMed

Bhattacharya, Munna; Biswas, Dipa; Sana, Santanu; Datta, Sriparna

2015-10-01

A potential degrader of paraffinic and aromatic hydrocarbons was isolated from oil-contaminated soil from steel plant effluent area in Burnpur, India. The strain was investigated for degradation of waste lubricants (waste engine oil and waste transformer oil) that often contain EPA (Environmental Protection Agency, USA) classified priority pollutants and was identified as Ochrobactrum sp. C1 by 16S rRNA gene sequencing. The strain C1 was found to tolerate unusually high waste lubricant concentration along with emulsification capability of the culture broth, and its degradation efficiency was 48.5 ± 0.5 % for waste engine oil and 30.47 ± 0.25 % for waste transformer oil during 7 days incubation period. In order to get optimal degradation efficiency, a three level Box-Behnken design was employed to optimize the physical parameters namely pH, temperature and waste oil concentration. The results indicate that at temperature 36.4 °C, pH 7.3 and with 4.6 % (v/v) oil concentration, the percentage degradation of waste engine oil will be 57 % within 7 days. At this optimized condition, the experimental values (56.7 ± 0.25 %) are in a good agreement with the predicted values with a calculated R 2 to be 0.998 and significant correlation between biodegradation and emulsification activity (E 24  = 69.42 ± 0.32 %) of the culture broth toward engine oil was found with a correlation coefficient of 0.972. This is the first study showing that an Ochrobactrum sp. strain is capable of degrading waste lubricants, which might contribute to the bioremediation of waste lubricating oil-contaminated soil.

Knowledge, Attitude and Practice of Healthcare Managers to Medical Waste Management and Occupational Safety Practices: Findings from Southeast Nigeria.

PubMed

Anozie, Okechukwu Bonaventure; Lawani, Lucky Osaheni; Eze, Justus Ndulue; Mamah, Emmanuel Johnbosco; Onoh, Robinson Chukwudi; Ogah, Emeka Onwe; Umezurike, Daniel Akuma; Anozie, Rita Onyinyechi

2017-03-01

Awareness of appropriate waste management procedures and occupational safety measures is fundamental to achieving a safe work environment, and ensuring patient and staff safety. This study was conducted to assess the attitude of healthcare managers to medical waste management and occupational safety practices. This was a cross-sectional study conducted among 54 hospital administrators in Ebonyi state. Semi-structured questionnaires were used for qualitative data collection and analyzed with SPSS statistics for windows (2011), version 20.0 statistical software (Armonk, NY: IBM Corp). Two-fifth (40%) of healthcare managers had received training on medical waste management and occupational safety. Standard operating procedure of waste disposal was practiced by only one hospital (1.9%), while 98.1% (53/54) practiced indiscriminate waste disposal. Injection safety boxes were widely available in all health facilities, nevertheless, the use of incinerators and waste treatment was practiced by 1.9% (1/54) facility. However, 40.7% (22/54) and 59.3% (32/54) of respondents trained their staff and organize safety orientation courses respectively. Staff insurance cover was offered by just one hospital (1.9%), while none of the hospitals had compensation package for occupational hazard victims. Over half (55.6%; 30/54) of the respondents provided both personal protective equipment and post exposure prophylaxis for HIV. There was high level of non-compliance to standard medical waste management procedures, and lack of training on occupational safety measures. Relevant regulating agencies should step up efforts at monitoring and regulation of healthcare activities and ensure staff training on safe handling and disposal of hospital waste.

Extraction of medium chain fatty acids from organic municipal waste and subsequent production of bio-based fuels.

PubMed

Kannengiesser, Jan; Sakaguchi-Söder, Kaori; Mrukwia, Timo; Jager, Johannes; Schebek, Liselotte

2016-01-01

This paper provides an overview on investigations for a new technology to generate bio-based fuel additives from bio-waste. The investigations are taking place at the composting plant in Darmstadt-Kranichstein (Germany). The aim is to explore the potential of bio-waste as feedstock in producing different bio-based products (or bio-based fuels). For this investigation, a facultative anaerobic process is to be integrated into the normal aerobic waste treatment process for composting. The bio-waste is to be treated in four steps to produce biofuels. The first step is the facultative anaerobic treatment of the waste in a rotting box namely percolate to generate a fatty-acid rich liquid fraction. The Hydrolysis takes place in the rotting box during the waste treatment. The organic compounds are then dissolved and transferred into the waste liquid phase. Browne et al. (2013) describes the hydrolysis as an enzymatically degradation of high solid substrates to soluble products which are further degraded to volatile fatty acids (VFA). This is confirmed by analytical tests done on the liquid fraction. After the percolation, volatile and medium chain fatty acids are found in the liquid phase. Concentrations of fatty acids between 8.0 and 31.5 were detected depending on the nature of the input material. In the second step, a fermentation process will be initiated to produce additional fatty acids. Existing microorganism mass is activated to degrade the organic components that are still remaining in the percolate. After fermentation the quantity of fatty acids in four investigated reactors increased 3-5 times. While fermentation mainly non-polar fatty acids (pentanoic to octanoic acid) are build. Next to the fermentation process, a chain-elongation step is arranged by adding ethanol to the fatty acid rich percolate. While these investigations a chain-elongation of mainly fatty acids with pair numbers of carbon atoms (acetate, butanoic and hexanoic acid) are demonstrated. After these three pre-treatments, the percolate is brought to a refinery to extract the non-polar fatty acids using bio-diesel, which was generated from used kitchen oil at the refinery. The extraction tests in the lab have proved that the efficiency of the liquid-liquid-extraction is directly linked with the chain length and polarity of the fatty acids. By using a non-polar bio-diesel mainly the non-polar fatty acids, like pentanoic to octanoic acid, are extracted. After extraction, the bio-diesel enriched with the fatty acids is esterified. As a result bio-diesel with a lower viscosity than usual is produced. The fatty acids remaining in the percolate after the extraction can be used in another fermentation process to generate biogas. Copyright © 2015 Elsevier Ltd. All rights reserved.

Engineering study for a melting, casting, rolling and fabrication facility for recycled contaminated stainless steel

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

This Preliminary Report is prepared to study the facilities required for recycling contaminated stainless steel scrap into plate which will be fabricated into boxes suitable for the storage of contaminated wastes and rubble. The study is based upon the underlying premise that the most cost effective way to produce stainless steel is to use the same processes employed by companies now in production of high quality stainless steel. Therefore, the method selected for this study for the production of stainless steel plate from scrap is conventional process using an Electric Arc Furnace for meltdown to hot metal, a Continuous Castermore » for production of cast slabs, and a Reversing Hot Mill for rolling the slabs into plate. The fabrication of boxes from the plate utilizes standard Shears, Punch Presses and welding equipment with Robotic Manipulators. This Study presumes that all process fumes, building dusts and vapors will be cycled through a baghouse and a nuclear grade HEPA filter facility prior to discharge. Also, all process waste water will be evaporated into the hot flue gas stream from the furnace utilizing a quench tank; so there will be no liquid discharges from the facility and all vapors will be processed through a HEPA filter. Even though HEPA filters are used today in controlling radioactive contamination from nuclear facilities there is a sparsity of data concerning radioactivity levels and composition of waste that may be collected from contaminated scrap steel processing. This report suggests some solutions to these problems but it is recommended that additional study must be given to these environmental problems.« less

Biotin-streptavidin enzyme-linked immunosorbent assay for detecting Tetrabromobisphenol A in electronic waste.

PubMed

Bu, Dan; Zhuang, Huisheng; Zhou, Xinchu; Yang, Guangxin

2014-03-01

Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant. A sensitive and selective indirect competitive biotin-streptavidin-amplified enzyme-linked immunosorbent assay (BA-ELISA) was developed for detecting TBBPA. The optimal hapten of TBBPA was 2-(2,6-dibromo-4-(2-(3,5-dibromo-4-hydroxyphenly)propan-2-yl)) acetic acid. Several physiochemical factors that influence assay performance, such as optimal coupling concentration of immunogen and antibody, organic solvent, ionic strength, and pH, were studied and optimized. The limit of detection (IC10) was 0.027 ng/mL and the median inhibitory concentration (IC50) was 0.58 ng/mL. The BA-ELISA was highly selective, with low cross-reactivity with TBBPA analogs. Finally, the assay was used to detect TBBPA in electronic waste samples. The results are consistent with those using liquid chromatography, which proves that the proposed immunoassay is accurate and receptive. This BA-ELISA method is suitable for the rapid and sensitive screening of TBBPA in environmental monitoring. © 2013 Published by Elsevier B.V.

Atrogin-1, a muscle-specific F-box protein highly expressed during muscle atrophy

NASA Technical Reports Server (NTRS)

Gomes, M. D.; Lecker, S. H.; Jagoe, R. T.; Navon, A.; Goldberg, A. L.

2001-01-01

Muscle wasting is a debilitating consequence of fasting, inactivity, cancer, and other systemic diseases that results primarily from accelerated protein degradation by the ubiquitin-proteasome pathway. To identify key factors in this process, we have used cDNA microarrays to compare normal and atrophying muscles and found a unique gene fragment that is induced more than ninefold in muscles of fasted mice. We cloned this gene, which is expressed specifically in striated muscles. Because this mRNA also markedly increases in muscles atrophying because of diabetes, cancer, and renal failure, we named it atrogin-1. It contains a functional F-box domain that binds to Skp1 and thereby to Roc1 and Cul1, the other components of SCF-type Ub-protein ligases (E3s), as well as a nuclear localization sequence and PDZ-binding domain. On fasting, atrogin-1 mRNA levels increase specifically in skeletal muscle and before atrophy occurs. Atrogin-1 is one of the few examples of an F-box protein or Ub-protein ligase (E3) expressed in a tissue-specific manner and appears to be a critical component in the enhanced proteolysis leading to muscle atrophy in diverse diseases.

Data sharing report characterization of population 7: Personal protective equipment, dry active waste, and miscellaneous debris, surveillance and maintenance project Oak Ridge National Laboratory Oak Ridge, Tennessee

DOE Office of Scientific and Technical Information (OSTI.GOV)

Harpenau, Evan M.

2013-10-10

The U.S. Department of Energy (DOE) Oak Ridge Office of Environmental Management (EM-OR) requested that Oak Ridge Associated Universities (ORAU), working under the Oak Ridge Institute for Science and Education (ORISE) contract, provide technical and independent waste management planning support under the American Recovery and Reinvestment Act (ARRA). Specifically, DOE EM-OR requested that ORAU plan and implement a sampling and analysis campaign targeting certain URS|CH2M Oak Ridge, LLC (UCOR) surveillance and maintenance (S&M) process inventory waste. Eight populations of historical and reoccurring S&M waste at the Oak Ridge National Laboratory (ORNL) have been identified in the Waste Handling Plan formore » Surveillance and Maintenance Activities at the Oak Ridge National Laboratory, DOE/OR/01-2565&D2 (WHP) (DOE 2012) for evaluation and processing to determine a final pathway for disposal. Population 7 (POP 7) consists of 56 containers of aged, low-level and potentially mixed S&M waste that has been staged in various locations around ORNL. Several of these POP 7 containers primarily contain personal protective equipment (PPE) and dry active waste (DAW), but may contain other miscellaneous debris. This data sharing report addresses the Comprehensive Environmental Response, Compensation, and Liability Act of 1980 (CERCLA) specified waste in a 13-container subpopulation (including eight steel boxes, three 55-gal drums, one sealand, and one intermodal) that lacked sufficient characterization data for possible disposal at the Environmental Management Waste Management Facility (EMWMF) using the approved Waste Lot (WL) 108.1 profile.« less

Evaluating the toxicity of food processing wastes as co-digestion substrates with dairy manure.

PubMed

Lisboa, Maria Sol; Lansing, Stephanie

2014-07-01

Studies have shown that including food waste as a co-digestion substrate in the anaerobic digestion of livestock manure can increase energy production. However, the type and inclusion rate of food waste used for co-digestion need to be carefully considered in order to prevent adverse conditions in the digestion environment. This study determined the effect of increasing the concentration (2%, 5%, 15% and 30%, by volume) of four food-processing wastes (meatball, chicken, cranberry and ice cream processing wastes) on methane production. Anaerobic toxicity assay (ATA) and specific methanogenic activity (SMA) tests were conducted to determine the concentration at which each food waste became toxic to the digestion environment. Decreases in methane production were observed at concentrations above 5% for all four food waste substrates, with up to 99% decreases in methane production at 30% food processing wastes (by volume). Copyright © 2014 Elsevier Ltd. All rights reserved.

Thermal and enzymatic recovering of proteins from untanned leather waste.

PubMed

Bajza, Z; Vrucek, V

2001-01-01

The laboratory trials of a process to treat untanned leather waste to isolate valuable protein products are presented. In this comparative study, both thermal and enzymatic treatments of leather waste were performed. The enzymatic method utilizes commercially available alkaline protease at moderate temperatures and for short periods of time. The concentration of the enzyme was 500 units per gram of leather waste which makes the method cost-effective. Amino acid composition in the hydrolysate obtained by the enzyme hydrolysis of untanned leather waste is determined. Chemical and physical properties of protein powder products from untanned leather waste were evaluated by spectrophotometric and chromatographic methods and by use of electron microscope. The results of microbiological assays confirm that these products agree to food safety standards. This relatively simple treatment of untanned leather waste may provide a practical and economical solution to the disposal of potentially dangerous waste.

Radiological Characterization Methodology of INEEL Stored RH-TRU Waste from ANL-E

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rajiv N. Bhatt

2003-02-01

An Acceptable Knowledge (AK)-based radiological characterization methodology is being developed for RH TRU waste generated from ANL-E hot cell operations performed on fuel elements irradiated in the EBR-II reactor. The methodology relies on AK for composition of the fresh fuel elements, their irradiation history, and the waste generation and collection processes. Radiological characterization of the waste involves the estimates of the quantities of significant fission products and transuranic isotopes in the waste. Methods based on reactor and physics principles are used to achieve these estimates. Because of the availability of AK and the robustness of the calculation methods, the AK-basedmore » characterization methodology offers a superior alternative to traditional waste assay techniques. Using this methodology, it is shown that the radiological parameters of a test batch of ANL-E waste is well within the proposed WIPP Waste Acceptance Criteria limits.« less

Radiological Characterization Methodology for INEEL-Stored Remote-Handled Transuranic (RH TRU) Waste from Argonne National Laboratory-East

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuan, P.; Bhatt, R.N.

2003-01-14

An Acceptable Knowledge (AK)-based radiological characterization methodology is being developed for RH TRU waste generated from ANL-E hot cell operations performed on fuel elements irradiated in the EBR-II reactor. The methodology relies on AK for composition of the fresh fuel elements, their irradiation history, and the waste generation and collection processes. Radiological characterization of the waste involves the estimates of the quantities of significant fission products and transuranic isotopes in the waste. Methods based on reactor and physics principles are used to achieve these estimates. Because of the availability of AK and the robustness of the calculation methods, the AK-basedmore » characterization methodology offers a superior alternative to traditional waste assay techniques. Using the methodology, it is shown that the radiological parameters of a test batch of ANL-E waste is well within the proposed WIPP Waste Acceptance Criteria limits.« less

Fully Automated RNAscope In Situ Hybridization Assays for Formalin‐Fixed Paraffin‐Embedded Cells and Tissues

PubMed Central

Anderson, Courtney M.; Zhang, Bingqing; Miller, Melanie; Butko, Emerald; Wu, Xingyong; Laver, Thomas; Kernag, Casey; Kim, Jeffrey; Luo, Yuling; Lamparski, Henry; Park, Emily; Su, Nan

2016-01-01

ABSTRACT Biomarkers such as DNA, RNA, and protein are powerful tools in clinical diagnostics and therapeutic development for many diseases. Identifying RNA expression at the single cell level within the morphological context by RNA in situ hybridization provides a great deal of information on gene expression changes over conventional techniques that analyze bulk tissue, yet widespread use of this technique in the clinical setting has been hampered by the dearth of automated RNA ISH assays. Here we present an automated version of the RNA ISH technology RNAscope that is adaptable to multiple automation platforms. The automated RNAscope assay yields a high signal‐to‐noise ratio with little to no background staining and results comparable to the manual assay. In addition, the automated duplex RNAscope assay was able to detect two biomarkers simultaneously. Lastly, assay consistency and reproducibility were confirmed by quantification of TATA‐box binding protein (TBP) mRNA signals across multiple lots and multiple experiments. Taken together, the data presented in this study demonstrate that the automated RNAscope technology is a high performance RNA ISH assay with broad applicability in biomarker research and diagnostic assay development. J. Cell. Biochem. 117: 2201–2208, 2016. © 2016 The Authors. Journal of Cellular Biochemistry Published by Wiley Periodicals, Inc. PMID:27191821

Alternative Radiological Characterization of Sealed Source TRU Waste for WIPP Disposal (LAUR-05-8776)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Whitworth, J.; Pearson, M.; Feldman, A.

2006-07-01

The Offsite Source Recovery (OSR) Project at Los Alamos National Laboratory is now shipping transuranic (TRU) waste containers to the Waste Isolation Pilot Plant (WIPP) in New Mexico for disposal. Sealed source waste disposal has become possible in part because OSR personnel were able to obtain Environmental Protection Agency (EPA) and DOE-CBFO approval for an alternative radiological characterization procedure relying on acceptable knowledge (AK) and modeling, rather than on non-destructive assay (NDA) of each container. This is the first successful qualification of an 'alternate methodology' under the radiological characterization requirements of the WIPP Waste Acceptance Criteria (WAC) by any TRUmore » waste generator site. This paper describes the approach OSR uses to radiologically characterize its sealed source waste and the process by which it obtained certification of this approach. (authors)« less

Size distributions of manure particles released under simulated rainfall.

PubMed

Pachepsky, Yakov A; Guber, Andrey K; Shelton, Daniel R; McCarty, Gregory W

2009-03-01

Manure and animal waste deposited on cropland and grazing lands serve as a source of microorganisms, some of which may be pathogenic. These microorganisms are released along with particles of dissolved manure during rainfall events. Relatively little if anything is known about the amounts and sizes of manure particles released during rainfall, that subsequently may serve as carriers, abode, and nutritional source for microorganisms. The objective of this work was to obtain and present the first experimental data on sizes of bovine manure particles released to runoff during simulated rainfall and leached through soil during subsequent infiltration. Experiments were conducted using 200 cm long boxes containing turfgrass soil sod; the boxes were designed so that rates of manure dissolution and subsequent infiltration and runoff could be monitored independently. Dairy manure was applied on the upper portion of boxes. Simulated rainfall (ca. 32.4 mm h(-1)) was applied for 90 min on boxes with stands of either live or dead grass. Electrical conductivity, turbidity, and particle size distributions obtained from laser diffractometry were determined in manure runoff and soil leachate samples. Turbidity of leachates and manure runoff samples decreased exponentially. Turbidity of manure runoff samples was on average 20% less than turbidity of soil leachate samples. Turbidity of leachate samples from boxes with dead grass was on average 30% less than from boxes with live grass. Particle size distributions in manure runoff and leachate suspensions remained remarkably stable after 15 min of runoff initiation, although the turbidity continued to decrease. Particles had the median diameter of 3.8 microm, and 90% of particles were between 0.6 and 17.8 microm. The particle size distributions were not affected by the grass status. Because manure particles are known to affect transport and retention of microbial pathogens in soil, more information needs to be collected about the concurrent release of pathogens and manure particles during rainfall events.

Biochemical methane potential (BMP) of agro-food wastes from the Cider Region (Spain).

PubMed

Nieto, P P; Hidalgo, D; Irusta, R; Kraut, D

2012-01-01

An inventory of agro-food industry organic waste streams with a high potential for biogas transformation was studied in a logistically viable area (Cider Region, Asturias, Spain). Three industries were selected as the most viable ones: livestock, dairy and beverage. The potential for methane production from six wastes (beverage waste, BW; milled apple waste, MA; milk waste, MK; yogurt waste, YG; fats and oils from dairy wastewater treatment, F&O and cattle manure, CM) at five different substrate:inoculum ratios (0.25, 0.50, 0.75, 1.00 and 1.50) was evaluated in laboratory batch assays. Obtained methane yields ranged from 202-549 mL STP CH(4)·g VS waste(-1), and the methane content in biogas ranged from 58-76%. The ultimate practical biochemical methane potentials were slightly affected by the substrate:inoculum ratio. The estimation of the regional fluxes of waste and methane potentials suggests anaerobic digestion as a sustainable solution for the valorization of the organic wastes generated in this Region.

Research on denitrification efficiency of three types of solid carbon source

NASA Astrophysics Data System (ADS)

Cai, Y.; Zhang, J. D.; Li, F.; Cao, Y. X.; Zhu, L. Y.; Xiao, M. S.

2018-01-01

C/N rates can greatly influence efficiency of denitrification. It is difficult for current treated effluent to reach GB18918-2002 primary effluent standard because of its low C/N rate. To improve the efficiency of denitrification, the quality of effluent, and realize the waste recycling, this article selected magnolia leaves, loofah and degradable meal box as the solid carbon source and set different solid-liquid ratio of magnolia leaves for periodic denitrification stage to study the change of NO3 --N, TN, COD, NO2 --N, NH4 +, PO4 3- and color. The results showed that in the condition of influent nitrate concentration of 40 mg/L, carbon dosage of 10 g, the reaction temperature of 25°C, the nitrate removal rates of magnolia leaves and loofah reached 89.0% and 96.8% respectively, rather higher than degradable meal box (56.3%). The TN removal rates of magnolia leaves (91.7%) and loofah (77.7%) were both higher than degradable meal box (53.9%), and the effluent TN concentration of loofah and degradable meal box reached 25.4 mg/L and 21.1 mg/L respectively, which couldn’t be discharged according to the primary effluent concentration standard of GB18918-2002. The released concentration of ammonia nitrogen and phosphate: loofah> magnolia> degradable meal box. The high solid-liquid ratio of magnolia leaves helped to improve the TN removal rate, which reached 75.0% (1:200) and 91.7% (1:100), but it caused higher released concentration of carbon, ammonia nitrogen and phosphate to effect system heavily. Under the integrated analysis, the low solid-liquid ratio (1:200) of magnolia leaves was more suitable to be the denitrification external carbon source.

Healthcare waste generation and management practice in government health centers of Addis Ababa, Ethiopia.

PubMed

Tadesse, Menelik Legesse; Kumie, Abera

2014-11-25

Healthcare wastes are hazardous organic and inorganic wastes. The waste disposal management in Addis Ababa city is seen unscientific manner. The waste management practice in the health facilities are poor and need improvement. This study will help different organizations, stakeholders and policy makers to correct and improve the existing situation of healthcare waste legislation and enforcement and training of staff in the healthcare facilities in Addis Ababa. The study aimed to assess the existing generation and management practice of healthcare waste in selected government health centers of Addis Ababa. The cross-sectional study was conducted to quantify waste generation rate and evaluate its management system. The study area was Addis Ababa. The sample size was determined by simple random sampling technique, the sampling procedure involved 10 sub-cities of Addis Ababa. Data were collected using both waste collecting and measuring equipment and check list. The Data was entered by EPI INFO version 6.04d and analyzed by and SPSS for WINDOW version15. The mean (±SD) healthcare waste generation rate was 9.61 ± 3.28 kg/day of which (38%) 3.64 ± 1.45 kg/day was general or non-hazardous waste and (62%) 5.97 ± 2.31 kg/day was hazardous. The mean healthcare waste generation rate between health centers was a significant different with Kurskal-Wallis test (χ2 = 21.83, p-value = 0.009). All health centers used safety boxes for collection of sharp wastes and all health centers used plastic buckets without lid for collection and transportation of healthcare waste. Pre treatment of infectious wastes was not practiced by any of the health centers. All health centers used incinerators and had placenta pit for disposal of pathological waste however only seven out of ten pits had proper covering material. Segregation of wastes at point of generation with appropriate collection materials and pre- treatment of infectious waste before disposal should be practiced. Training should be given to healthcare workers and waste handlers. Incinerators must be constructed in a manner that facilitates complete combustion and the lining of placenta pit should be constructed in water tight material.

A minimal murine Msx-1 gene promoter. Organization of its cis-regulatory motifs and their role in transcriptional activation in cells in culture and in transgenic mice.

PubMed

Takahashi, T; Guron, C; Shetty, S; Matsui, H; Raghow, R

1997-09-05

To dissect the cis-regulatory elements of the murine Msx-1 promoter, which lacks a conventional TATA element, a putative Msx-1 promoter DNA fragment (from -1282 to +106 base pairs (bp)) or its congeners containing site-specific alterations were fused to luciferase reporter and introduced into NIH3T3 and C2C12 cells, and the expression of luciferase was assessed in transient expression assays. The functional consequences of the sequential 5' deletions of the promotor revealed that multiple positive and negative regulatory elements participate in regulating transcription of the Msx-1 gene. Surprisingly, however, the optimal expression of Msx-1 promoter in either NIH3T3 or C2C12 cells required only 165 bp of the upstream sequence to warrant detailed examination of its structure. Therefore, the functional consequences of site-specific deletions and point mutations of the cis-acting elements of the minimal Msx-1 promoter were systematically examined. Concomitantly, potential transcriptional factor(s) interacting with the cis-acting elements of the minimal promoter were also studied by gel electrophoretic mobility shift assays and DNase I footprinting. Combined analyses of the minimal promoter by DNase I footprinting, electrophoretic mobility shift assays, and super shift assays with specific antibodies revealed that 5'-flanking regions from -161 to -154 and from -26 to -13 of the Msx-1 promoter contains an authentic E box (proximal E box), capable of binding a protein immunologically related to the upstream stimulating factor 1 (USF-1) and a GC-rich sequence motif which can bind to Sp1 (proximal Sp1), respectively. Additionally, we observed that the promoter activation was seriously hampered if the proximal E box was removed or mutated, and the promoter activity was eliminated completely if the proximal Sp1 site was similarly altered. Absolute dependence of the Msx-1 minimal promoter on Sp1 could be demonstrated by transient expression assays in the Sp1-deficient Drosophila cell line cotransfected with Msx-1-luciferase and an Sp1 expression vector pPacSp1. The transgenic mice embryos containing -165/106-bp Msx-1 promoter-LacZ DNA in their genomes abundantly expressed beta-galactosidase in maxillae and mandibles and in the cellular primordia involved in the formation of the meninges and the bones of the skull. Thus, the truncated murine Msx-1 promoter can target expression of a heterologous gene in the craniofacial tissues of transgenic embryos known for high level of expression of the endogenous Msx-1 gene and found to be severely defective in the Msx-1 knock-out mice.

A Procedure for Evaluating Subpotable Water Reuse Potential at Army Fixed Facilities.

DTIC Science & Technology

1981-11-01

characteristic of phosphate waste waters from fume scrubbers. + Flue gas desulfurization sludge liquors. 75 Table B5 Base Housing Water Supply Quality (Also...ENGINEERING RESEARCH LABORATORY 4A762720A896-B-008 P.O. Box 4005, Champaign, IL 61820 1I. CONTROLLING OFFICE NAME AND ADDRESS 12. REPORT DATE November 1981 13...NUMBER OF PAGES 102 4. MONITORING AGENCY NAME & ADDRESS(If different from Controlling Office) IS. SECURITY CLASS. (of til, report) UNCLASSIFIED 15

VIEW OF THE INTERIOR OF BUILDING 774, THE ORIGINAL LIQUID ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

VIEW OF THE INTERIOR OF BUILDING 774, THE ORIGINAL LIQUID PROCESS WASTEWATER TREATMENT FACILITY. THE PHOTOGRAPH SHOWS STORAGE TANKS AND ASSOCIATED PLUTONIUM-CONTAMINATED SOLUTIONS. THE GLOVE BOX IS USED BY OPERATORS TO MANUALLY OPERATE PUMPS AND VALVES THAT REQUIRE PERIODIC ADJUSTMENT. OTHER VALVES IN THE ROOM WERE INFREQUENTLY ADJUSTED, AND ARE SEALED IN PLASTIC WRAP - Rocky Flats Plant, Waste Treatment Facility, Adjacent to bldg 771C, in northern portion of protected area, Golden, Jefferson County, CO

Wheat Chloroplast Targeted sHSP26 Promoter Confers Heat and Abiotic Stress Inducible Expression in Transgenic Arabidopsis Plants

PubMed Central

Khurana, Neetika; Chauhan, Harsh; Khurana, Paramjit

2013-01-01

The small heat shock proteins (sHSPs) have been found to play a critical role in physiological stress conditions in protecting proteins from irreversible aggregation. To characterize the hloroplast targeted sHSP26 promoter in detail, deletion analysis of the promoter is carried out and analysed via transgenics in Arabidopsis. In the present study, complete assessment of the importance of CCAAT-box elements along with Heat shock elements (HSEs) in the promoter of sHSP26 was performed. Moreover, the importance of 5′ untranslated region (UTR) has also been established in the promoter via Arabidopsis transgenics. An intense GUS expression was observed after heat stress in the transgenics harbouring a full-length promoter, confirming the heat-stress inducibility of the promoter. Transgenic plants without UTR showed reduced GUS expression when compared to transgenic plants with UTR as was confirmed at the RNA and protein levels by qRT-PCR and GUS histochemical assays, thus suggesting the possible involvement of some regulatory elements present in the UTR in heat-stress inducibility of the promoter. Promoter activity was also checked under different abiotic stresses and revealed differential expression in different deletion constructs. Promoter analysis based on histochemical assay, real-time qPCR and fluorimetric analysis revealed that HSEs alone could not transcribe GUS gene significantly in sHSP26 promoter and CCAAT box elements contribute synergistically to the transcription. Our results also provide insight into the importance of 5`UTR of sHsp26 promoter thus emphasizing the probable role of imperfect CCAAT-box element or some novel cis-element with respect to heat stress. PMID:23349883

The integrated endoplasmic reticulum stress response in Leishmania amazonensis macrophage infection: the role of X-box binding protein 1 transcription factor.

PubMed

Dias-Teixeira, Karina Luiza; Calegari-Silva, Teresa Cristina; dos Santos, Guilherme R R M; Vitorino Dos Santos, José; Lima, Carolina; Medina, Jorge Mansur; Aktas, Bertal Huseyin; Lopes, Ulisses G

2016-04-01

Endoplasmic reticulum (ER) stress triggers the integrated ER-stress response (IERSR) that ensures cellular survival of ER stress and represents a primordial form of innate immunity. We investigated the role of IERSR duringLeishmania amazonensisinfection. Treatment of RAW 264.7 infected macrophages with the ER stress-inducing agent thapsigargin (TG; 1 μM) increasedL. amazonensisinfectivity in an IFN1-α receptor (IFNAR)-dependent manner. In Western blot assays, we showed thatL. amazonensisactivates the inositol-requiring enzyme (IRE1)/ X-box binding protein (XBP)-1-splicing arms of the IERSR in host cells. In chromatin immunoprecipitation (ChIP) assays, we showed an increased occupancy of enhancer and promoter sequences for theIfnbgene by XBP1 in infected RAW 264.7 cells. Knocking down XBP1 expression by transducing RAW 264.7 cells with the short hairpin XBP1 lentiviral vector significantly reduced the parasite proliferation associated with impaired translocation of phosphorylated IFN regulatory transcription factor (IRF)-3 to the nucleus and a decrease in IFN1-β expression. Knocking down XBP1 expression also increased NO concentration, as determined by Griess reaction and reduced the expression of antioxidant genes, such as heme oxygenase (HO)-1, that protect parasites from oxidative stress. We conclude thatL. amazonensisactivation of XBP1 plays a critical role in infection by protecting the parasites from oxidative stress and increasing IFN1-β expression.-Dias-Teixeira, K. L., Calegari-Silva, T. C., Dos Santos, G. R. R. M., Vitorino dos Santos, J., Lima, C., Medina, J. M., Aktas, B. H., Lopes, U. G. The integrated endoplasmic reticulum stress response inLeishmania amazonensismacrophage infection: the role of X-box binding protein 1 transcription factor. © FASEB.

Bremsstrahlung-Based Imaging and Assays of Radioactive, Mixed and Hazardous Waste

NASA Astrophysics Data System (ADS)

Kwofie, J.; Wells, D. P.; Selim, F. A.; Harmon, F.; Duttagupta, S. P.; Jones, J. L.; White, T.; Roney, T.

2003-08-01

A new nondestructive accelerator based x-ray fluorescence (AXRF) approach has been developed to identify heavy metals in large-volume samples. Such samples are an important part of the process and waste streams of U.S Department of Energy sites, as well as other industries such as mining and milling. Distributions of heavy metal impurities in these process and waste samples can range from homogeneous to highly inhomogeneous, and non-destructive assays and imaging that can address both are urgently needed. Our approach is based on using high-energy, pulsed bremsstrahlung beams (3-6.5 MeV) from small electron accelerators to produce K-shell atomic fluorescence x-rays. In addition we exploit pair-production, Compton scattering and x-ray transmission measurements from these beams to probe locations of high density and high atomic number. The excellent penetrability of these beams allows assays and images for soil-like samples at least 15 g/cm2 thick, with elemental impurities of atomic number greater than approximately 50. Fluorescence yield of a variety of targets was measured as a function of impurity atomic number, impurity homogeneity, and sample thickness. We report on actual and potential detection limits of heavy metal impurities in a soil matrix for a variety of samples, and on the potential for imaging, using AXRF and these related probes.

Genome-wide identification of the MADS-box transcription factor family in pear (Pyrus bretschneideri) reveals evolution and functional divergence.

PubMed

Wang, Runze; Ming, Meiling; Li, Jiaming; Shi, Dongqing; Qiao, Xin; Li, Leiting; Zhang, Shaoling; Wu, Jun

2017-01-01

MADS-box transcription factors play significant roles in plant developmental processes such as floral organ conformation, flowering time, and fruit development. Pear ( Pyrus ), as the third-most crucial temperate fruit crop, has been fully sequenced. However, there is limited information about the MADS family and its functional divergence in pear. In this study, a total of 95 MADS-box genes were identified in the pear genome, and classified into two types by phylogenetic analysis. Type I MADS-box genes were divided into three subfamilies and type II genes into 14 subfamilies. Synteny analysis suggested that whole-genome duplications have played key roles in the expansion of the MADS family, followed by rearrangement events. Purifying selection was the primary force driving MADS-box gene evolution in pear, and one gene pairs presented three codon sites under positive selection. Full-scale expression information for PbrMADS genes in vegetative and reproductive organs was provided and proved by transcriptional and reverse transcription PCR analysis. Furthermore, the PbrMADS11(12) gene, together with partners PbMYB10 and PbbHLH3 was confirmed to activate the promoters of the structural genes in anthocyanin pathway of red pear through dual luciferase assay. In addition, the PbrMADS11 and PbrMADS12 were deduced involving in the regulation of anthocyanin synthesis response to light and temperature changes. These results provide a solid foundation for future functional analysis of PbrMADS genes in different biological processes, especially of pigmentation in pear.

Genome-wide identification of the MADS-box transcription factor family in pear (Pyrus bretschneideri) reveals evolution and functional divergence

PubMed Central

Li, Jiaming; Shi, Dongqing; Qiao, Xin; Li, Leiting; Zhang, Shaoling

2017-01-01

MADS-box transcription factors play significant roles in plant developmental processes such as floral organ conformation, flowering time, and fruit development. Pear (Pyrus), as the third-most crucial temperate fruit crop, has been fully sequenced. However, there is limited information about the MADS family and its functional divergence in pear. In this study, a total of 95 MADS-box genes were identified in the pear genome, and classified into two types by phylogenetic analysis. Type I MADS-box genes were divided into three subfamilies and type II genes into 14 subfamilies. Synteny analysis suggested that whole-genome duplications have played key roles in the expansion of the MADS family, followed by rearrangement events. Purifying selection was the primary force driving MADS-box gene evolution in pear, and one gene pairs presented three codon sites under positive selection. Full-scale expression information for PbrMADS genes in vegetative and reproductive organs was provided and proved by transcriptional and reverse transcription PCR analysis. Furthermore, the PbrMADS11(12) gene, together with partners PbMYB10 and PbbHLH3 was confirmed to activate the promoters of the structural genes in anthocyanin pathway of red pear through dual luciferase assay. In addition, the PbrMADS11 and PbrMADS12 were deduced involving in the regulation of anthocyanin synthesis response to light and temperature changes. These results provide a solid foundation for future functional analysis of PbrMADS genes in different biological processes, especially of pigmentation in pear. PMID:28924499

SlDEAD31, a Putative DEAD-Box RNA Helicase Gene, Regulates Salt and Drought Tolerance and Stress-Related Genes in Tomato.

PubMed

Zhu, Mingku; Chen, Guoping; Dong, Tingting; Wang, Lingling; Zhang, Jianling; Zhao, Zhiping; Hu, Zongli

2015-01-01

The DEAD-box RNA helicases are involved in almost every aspect of RNA metabolism, associated with diverse cellular functions including plant growth and development, and their importance in response to biotic and abiotic stresses is only beginning to emerge. However, none of DEAD-box genes was well characterized in tomato so far. In this study, we reported on the identification and characterization of two putative DEAD-box RNA helicase genes, SlDEAD30 and SlDEAD31 from tomato, which were classified into stress-related DEAD-box proteins by phylogenetic analysis. Expression analysis indicated that SlDEAD30 was highly expressed in roots and mature leaves, while SlDEAD31 was constantly expressed in various tissues. Furthermore, the expression of both genes was induced mainly in roots under NaCl stress, and SlDEAD31 mRNA was also increased by heat, cold, and dehydration. In stress assays, transgenic tomato plants overexpressing SlDEAD31 exhibited dramatically enhanced salt tolerance and slightly improved drought resistance, which were simultaneously demonstrated by significantly enhanced expression of multiple biotic and abiotic stress-related genes, higher survival rate, relative water content (RWC) and chlorophyll content, and lower water loss rate and malondialdehyde (MDA) production compared to wild-type plants. Collectively, these results provide a preliminary characterization of SlDEAD30 and SlDEAD31 genes in tomato, and suggest that stress-responsive SlDEAD31 is essential for salt and drought tolerance and stress-related gene regulation in plants.

A novel in vitro image-based assay identifies new drug leads for giardiasis.

PubMed

Hart, Christopher J S; Munro, Taylah; Andrews, Katherine T; Ryan, John H; Riches, Andrew G; Skinner-Adams, Tina S

2017-04-01

Giardia duodenalis is an intestinal parasite that causes giardiasis, a widespread human gastrointestinal disease. Treatment of giardiasis relies on a small arsenal of compounds that can suffer from limitations including side-effects, variable treatment efficacy and parasite drug resistance. Thus new anti-Giardia drug leads are required. The search for new compounds with anti-Giardia activity currently depends on assays that can be labour-intensive, expensive and restricted to measuring activity at a single time-point. Here we describe a new in vitro assay to assess anti-Giardia activity. This image-based assay utilizes the Perkin-Elmer Operetta ® and permits automated assessment of parasite growth at multiple time points without cell-staining. Using this new approach, we assessed the "Malaria Box" compound set for anti-Giardia activity. Three compounds with sub-μM activity (IC 50 0.6-0.9 μM) were identified as potential starting points for giardiasis drug discovery. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Molecular Cloning of a cDNA Encoding for Taenia solium TATA-Box Binding Protein 1 (TsTBP1) and Study of Its Interactions with the TATA-Box of Actin 5 and Typical 2-Cys Peroxiredoxin Genes

PubMed Central

Rodríguez-Lima, Oscar; García-Gutierrez, Ponciano; Jiménez, Lucía; Zarain-Herzberg, Ángel; Lazzarini, Roberto; Landa, Abraham

2015-01-01

TATA-box binding protein (TBP) is an essential regulatory transcription factor for the TATA-box and TATA-box-less gene promoters. We report the cloning and characterization of a full-length cDNA that encodes a Taenia solium TATA-box binding protein 1 (TsTBP1). Deduced amino acid composition from its nucleotide sequence revealed that encodes a protein of 238 residues with a predicted molecular weight of 26.7 kDa, and a theoretical pI of 10.6. The NH2-terminal domain shows no conservation when compared with to pig and human TBP1s. However, it shows high conservation in size and amino acid identity with taeniids TBP1s. In contrast, the TsTBP1 COOH-terminal domain is highly conserved among organisms, and contains the amino acids involved in interactions with the TATA-box, as well as with TFIIA and TFIIB. In silico TsTBP1 modeling reveals that the COOH-terminal domain forms the classical saddle structure of the TBP family, with one α-helix at the end, not present in pig and human. Native TsTBP1 was detected in T. solium cysticerci´s nuclear extract by western blot using rabbit antibodies generated against two synthetic peptides located in the NH2 and COOH-terminal domains of TsTBP1. These antibodies, through immunofluorescence technique, identified the TBP1 in the nucleus of cells that form the bladder wall of cysticerci of Taenia crassiceps, an organism close related to T. solium. Electrophoretic mobility shift assays using nuclear extracts from T. solium cysticerci and antibodies against the NH2-terminal domain of TsTBP1 showed the interaction of native TsTBP1 with the TATA-box present in T. solium actin 5 (pAT5) and 2-Cys peroxiredoxin (Ts2-CysPrx) gene promoters; in contrast, when antibodies against the anti-COOH-terminal domain of TsTBP1 were used, they inhibited the binding of TsTBP1 to the TATA-box of the pAT5 promoter gene. PMID:26529408

A combined approach to investigate the toxicity of an industrial landfill's leachate: Chemical analyses, risk assessment and in vitro assays

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baderna, D., E-mail: diego.baderna@marionegri.it; Maggioni, S.; Boriani, E.

2011-05-15

Solid wastes constitute an important and emerging problem. Landfills are still one of the most common ways to manage waste disposal. The risk assessment of pollutants from landfills is becoming a major environmental issue in Europe, due to the large number of sites and to the importance of groundwater protection. Furthermore, there is lack of knowledge for the environmental, ecotoxicological and toxicological characteristics of most contaminants contained into landfill leacheates. Understanding leachate composition and creating an integrated strategy for risk assessment are currently needed to correctly face the landfill issues and to make projections on the long-term impacts of amore » landfill, with particular attention to the estimation of possible adverse effects on human health and ecosystem. In the present study, we propose an integrated strategy to evaluate the toxicity of the leachate using chemical analyses, risk assessment guidelines and in vitro assays using the hepatoma HepG2 cells as a model. The approach was applied on a real case study: an industrial waste landfill in northern Italy for which data on the presence of leachate contaminants are available from the last 11 years. Results from our ecological risk models suggest important toxic effects on freshwater fish and small rodents, mainly due to ammonia and inorganic constituents. Our results from in vitro data show an inhibition of cell proliferation by leachate at low doses and cytotoxic effect at high doses after 48 h of exposure. - Research highlights: {yields} We study the toxicity of leachate from a non-hazardous industrial waste landfill. {yields} We perform chemical analyses, risk assessments and in vitro assays on HepG2 cells. {yields} Risk models suggest toxic effects due to ammonia and inorganic constituents. {yields} In vitro assays show that leachate inhibits cell proliferation at low doses. {yields} Leachate can induce cytotoxic effects on HepG2 cells at high doses.« less

Medically relevant assays with a simple smartphone and tablet based fluorescence detection system.

PubMed

Wargocki, Piotr; Deng, Wei; Anwer, Ayad G; Goldys, Ewa M

2015-05-20

Cell phones and smart phones can be reconfigured as biomedical sensor devices but this requires specialized add-ons. In this paper we present a simple cell phone-based portable bioassay platform, which can be used with fluorescent assays in solution. The system consists of a tablet, a polarizer, a smart phone (camera) and a box that provides dark readout conditions. The assay in a well plate is placed on the tablet screen acting as an excitation source. A polarizer on top of the well plate separates excitation light from assay fluorescence emission enabling assay readout with a smartphone camera. The assay result is obtained by analysing the intensity of image pixels in an appropriate colour channel. With this device we carried out two assays, for collagenase and trypsin using fluorescein as the detected fluorophore. The results of collagenase assay with the lowest measured concentration of 3.75 µg/mL and 0.938 µg in total in the sample were comparable to those obtained by a microplate reader. The lowest measured amount of trypsin was 930 pg, which is comparable to the low detection limit of 400 pg for this assay obtained in a microplate reader. The device is sensitive enough to be used in point-of-care medical diagnostics of clinically relevant conditions, including arthritis, cystic fibrosis and acute pancreatitis.

Comparative evaluation of environmental contamination and DNA damage induced by electronic-waste in Nigeria and China.

PubMed

Alabi, Okunola A; Bakare, Adekunle A; Xu, Xijin; Li, Bin; Zhang, Yuling; Huo, Xia

2012-04-15

In the last decade, China and Nigeria have been prime destinations for the world's e-waste disposal leading to serious environmental contamination. We carried out a comparative study of the level of contamination using soils and plants from e-waste dumping and processing sites in both countries. Levels of polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were analyzed using gas chromatography/spectrophotometry and heavy metals using atomic absorption spectrophotometry. DNA damage was assayed in human peripheral blood lymphocytes using an alkaline comet assay. Soils and plants were highly contaminated with toxic PAHs, PCBs, PBDEs, and heavy metals in both countries. Soil samples from China and plant samples from Nigeria were more contaminated. There was a positive correlation between the concentrations of organics and heavy metals in plant samples and the surrounding soils. In human lymphocytes, all tested samples induced significant (p<0.05) concentration-dependent increases in DNA damage compared with the negative control. These findings suggest that e-waste components/constituents can accumulate, in soil and surrounding vegetation, to toxic and genotoxic levels that could induce adverse health effects in exposed individuals. Copyright © 2012 Elsevier B.V. All rights reserved.

Olympic boxing is associated with elevated levels of the neuronal protein tau in plasma.

PubMed

Neselius, Sanna; Zetterberg, Henrik; Blennow, Kaj; Randall, Jeffrey; Wilson, David; Marcusson, Jan; Brisby, Helena

2013-01-01

The aim of this study was to investigate if olympic (amateur) boxing is associated with elevation of brain injury biomarkers in peripheral blood compared to controls. Thirty olympic boxers competing in at least 47 bouts were compared to 25 controls. Blood was collected from the controls at one occasion and from the boxers within 1-6 days after a bout and after a rest period of at least 14 days. Tau concentration in plasma was determined using a novel single molecule ELISA assay and S-100B, glial fibrillary acidic protein, brain-derived neurotrophic factor and amyloid β 1-42 were determined using standard immunoassays. None of the boxers had been knocked-out during the bout. Plasma-tau was significantly increased in the boxers after a bout compared to controls (mean ± SD, 2.46 ± 5.10 vs. 0.79 ± 0.961 ng L(-1), p = 0.038). The other brain injury markers did not differ between the groups. Plasma-tau decreased significantly in the boxers after a resting period compared to after a bout (p = 0.030). Olympic boxing is associated with elevation of tau in plasma. The repetitive minimal head injury in boxing may lead to axonal injuries that can be diagnosed with a blood test.

Antioxidant and cytoprotective activities of extracts prepared from fruit and vegetable wastes and by-products.

PubMed

Kabir, Faisal; Tow, Wei Wei; Hamauzu, Yasunori; Katayama, Shigeru; Tanaka, Sachi; Nakamura, Soichiro

2015-01-15

In this study, fruit and vegetable wastes and by-products were tested for polyphenol content and their antioxidant activity. The highest content of polyphenols as assessed by the Folin-Ciocalteu assay was the hot-water extract of grape seed, followed by the ethanol extract of buckwheat hull. The highest antioxidant activity measured by 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assays was also detected in the hot-water extract of grape seed, followed by the ethanol extract of immature prune. Most of samples showed protective effects against oxidative stress induced by 2,2'-azobis-(2-amidinopropane) dihydrochloride (AAPH) peroxyl radical generator in African monkey kidney (MA 104) cells. Samples containing high amounts of phenolics (more than 30 mg ChAE/g) generally showed high antioxidant activity and a protective effect against AAPH-induced oxidative stress. This study demonstrates that fruit and vegetable wastes and by-products are good sources of high amounts of phenolics with antioxidant properties. Copyright © 2014 Elsevier Ltd. All rights reserved.

Molecular Characterization and Transcriptional Regulation Analysis of the Bovine PDHB Gene.

PubMed

Li, Anning; Zhang, Yaran; Zhao, Zhidong; Wang, Mingming; Zan, Linsen

2016-01-01

The pyruvate dehydrogenase beta subunit (PDHB) is a subunit of pyruvate dehydrogenase (E1), which catalyzes pyruvate into acetyl-CoA and provides a linkage between the tricarboxylic acid cycle (TCA) and the glycolysis pathway. Previous studies demonstrated PDHB to be positively related to the intramuscular fat (IMF) content. However, the transcriptional regulation of PDHB remains unclear. In our present study, the cDNA of bovine PDHB was cloned and the genomic structure was analyzed. The phylogenetic tree showed bovine PDHB to be closely related to goat and sheep, and least related to chicken. Spatial expression pattern analysis revealed the products of bovine PDHB to be widely expressed with the highest level in the fat of testis. To understand the transcriptional regulation of bovine PDHB, 1899 base pairs (bp) of the 5'-regulatory region was cloned. Sequence analysis neither found consensus TATA-box nor CCAAT-box in the 5'-flanking region of bovine PDHB. However, a CpG island was predicted from nucleotides -284 to +117. Serial deletion constructs of the 5'-flanking region, evaluated in dual-luciferase reporter assay, revealed the core promoter to be located 490bp upstream from the transcription initiation site (+1). Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP) in combination with asite-directed mutation experiment indicated both myogenin (MYOG) and the CCAAT/enhancer-binding protein beta (C/EBPß) to be important transcription factors for bovine PDHB in skeletal muscle cells and adipocytes. Our results provide an important basis for further investigation of the bovine PDHB function and regulation in cattle.

Identification of Four-Jointed Box 1 (FJX1)-Specific Peptides for Immunotherapy of Nasopharyngeal Carcinoma.

PubMed

Chai, San Jiun; Yap, Yoke Yeow; Foo, Yoke Ching; Yap, Lee Fah; Ponniah, Sathibalan; Teo, Soo Hwang; Cheong, Sok Ching; Patel, Vyomesh; Lim, Kue Peng

2015-01-01

Nasopharyngeal carcinoma (NPC) is highly prevalent in South East Asia and China. The poor outcome is due to late presentation, recurrence, distant metastasis and limited therapeutic options. For improved treatment outcome, immunotherapeutic approaches focusing on dendritic and autologous cytotoxic T-cell based therapies have been developed, but cost and infrastructure remain barriers for implementing these in low-resource settings. As our prior observations had found that four-jointed box 1 (FJX1), a tumor antigen, is overexpressed in NPCs, we investigated if short 9-20 amino acid sequence specific peptides matching to FJX1 requiring only intramuscular immunization to train host immune systems would be a better treatment option for this disease. Thus, we designed 8 FJX1-specific peptides and implemented an assay system to first, assess the binding of these peptides to HLA-A2 molecules on T2 cells. After, ELISPOT assays were used to determine the peptides immunogenicity and ability to induce potential cytotoxicity activity towards cancer cells. Also, T-cell proliferation assay was used to evaluate the potential of MHC class II peptides to stimulate the expansion of isolated T-cells. Our results demonstrate that these peptides are immunogenic and peptide stimulated T-cells were able to induce peptide-specific cytolytic activity specifically against FJX1-expressing cancer cells. In addition, we demonstrated that the MHC class II peptides were capable of inducing T-cell proliferation. Our results suggest that these peptides are capable of inducing specific cytotoxic cytokines secretion against FJX1-expressing cancer cells and serve as a potential vaccine-based therapy for NPC patients.

Molecular Characterization and Transcriptional Regulation Analysis of the Bovine PDHB Gene

PubMed Central

Li, Anning; Zhang, Yaran; Zhao, Zhidong; Wang, Mingming; Zan, Linsen

2016-01-01

The pyruvate dehydrogenase beta subunit (PDHB) is a subunit of pyruvate dehydrogenase (E1), which catalyzes pyruvate into acetyl-CoA and provides a linkage between the tricarboxylic acid cycle (TCA) and the glycolysis pathway. Previous studies demonstrated PDHB to be positively related to the intramuscular fat (IMF) content. However, the transcriptional regulation of PDHB remains unclear. In our present study, the cDNA of bovine PDHB was cloned and the genomic structure was analyzed. The phylogenetic tree showed bovine PDHB to be closely related to goat and sheep, and least related to chicken. Spatial expression pattern analysis revealed the products of bovine PDHB to be widely expressed with the highest level in the fat of testis. To understand the transcriptional regulation of bovine PDHB, 1899 base pairs (bp) of the 5’-regulatory region was cloned. Sequence analysis neither found consensus TATA-box nor CCAAT-box in the 5’-flanking region of bovine PDHB. However, a CpG island was predicted from nucleotides -284 to +117. Serial deletion constructs of the 5’-flanking region, evaluated in dual-luciferase reporter assay, revealed the core promoter to be located 490bp upstream from the transcription initiation site (+1). Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP) in combination with asite-directed mutation experiment indicated both myogenin (MYOG) and the CCAAT/enhancer-binding protein beta (C/EBPß) to be important transcription factors for bovine PDHB in skeletal muscle cells and adipocytes. Our results provide an important basis for further investigation of the bovine PDHB function and regulation in cattle. PMID:27379520

Evidence of the neuron-restrictive silencer factor (NRSF) interaction with Sp3 and its synergic repression to the mu opioid receptor (MOR) gene

PubMed Central

Kim, Chun Sung; Choi, Hack Sun; Hwang, Cheol Kyu; Song, Kyu Young; Lee, Byung-Kwon; Law, Ping-Yee; Wei, Li-Na; Loh, Horace H.

2006-01-01

Previously, we reported that the neuron-restrictive silencer element (NRSE) of mu opioid receptor (MOR) functions as a critical regulator to repress the MOR transcription in specific neuronal cells, depending on neuron-restriction silence factor (NRSF) expression levels [C.S.Kim, C.K.Hwang, H.S.Choi, K.Y.Song, P.Y.Law, L.N.Wei and H.H.Loh (2004) J. Biol. Chem., 279, 46464–46473]. Herein, we identify a conserved GC sequence next to NRSE region in the mouse MOR gene. The inhibition of Sp family factors binding to this GC box by mithramycin A led to a significant increase in the endogenous MOR transcription. In the co-immunoprecipitation experiment, NRSF interacted with the full-length Sp3 factor, but not with Sp1 or two short Sp3 isoforms. The sequence specific and functional binding by Sp3 at this GC box was confirmed by in vitro gel-shift assays using either in vitro translated proteins or nuclear extract, and by in vivo chromatin immunoprecipitation assays. Transient transfection assays showed that Sp3-binding site of the MOR gene is a functionally synergic repressor element with NRSE in NS20Y cells, but not in the NRSF negative PC12 cells. The results suggest that the synergic interaction between NRSF and Sp3 is required to negatively regulate MOR gene transcription and that transcription of MOR gene would be governed by the context of available transcription factors rather than by a master regulator. PMID:17130167

Asphalt dust waste material as a paste volume in developing sustainable self compacting concrete (SCC)

NASA Astrophysics Data System (ADS)

Ismail, Isham; Shahidan, Shahiron; Bahari, Nur Amira Afiza Saiful

2017-12-01

Self-compacting concrete (SCC) mixtures are usually designed to have high workability during the fresh state through the influence of higher volumes of paste in concrete mixtures. Asphalt dust waste (ADW) is one of disposed materials obtained during the production of asphalt premix. These fine powder wastes contribute to environmental problems today. However, these waste materials can be utilized in the development of sustainable and economical SCC. This paper focuses on the preliminary evaluations of the fresh properties and compressive strength of developed SCC for 7 and 28 days only. 144 cube samples from 24 mixtures with varying water binder ratios (0.2, 0.3 and 0.4) and ADW volume (0% to 100%) were prepared. MD940 and MD950 showed a satisfactory performance for the slump flow, J-Ring, L-Box and V-Funnel tests at fresh state. The compressive strength after 28 days for MD940 and MD950 was 36.9 MPa and 28.0 MPa respectively. In conclusion, the use of ADW as paste volume should be limited and a higher water binder ratio will significantly reduce the compressive strength.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gruetzmacher, Kathleen M.; Bustos, Roland M.; Ferran, Scott G.

Los Alamos National Laboratory (LANL) uses the Nevada National Security Site (NNSS) as an off-site disposal facility for low-level waste (LLW), including sludge waste. NNSS has issued a position paper that indicates that systems that are not certified by the Carlsbad Field Office (CBFO) for Waste Isolation Pilot Plant (WIPP) disposal of Transuranic (TRU) waste must demonstrate equivalent practices to the CBFO certified systems in order to assign activity concentration values to assayed items without adding in the Total Measurement Uncertainty (TMU) when certifying waste for NNSS disposal. Efforts have been made to meet NNSS requirements to accept sludge wastemore » for disposal at their facility. The LANL LLW Characterization Team uses portable high purity germanium (HPGe) detector systems for the nondestructive assay (NDA) of both debris and sludge LLW. A number of performance studies have been conducted historically by LANL to support the efficacy and quality of assay results generated by the LANL HPGe systems, and, while these detector systems are supported by these performance studies and used with LANL approved procedures and processes, they are not certified by CBFO for TRU waste disposal. Beginning in 2009, the LANL LLW Characterization Team undertook additional NDA measurements of both debris and sludge simulated waste containers to supplement existing studies and procedures to demonstrate full compliance with the NNSS position paper. Where possible, Performance Demonstration Project (PDP) drums were used for the waste matrix and PDP sources were used for the radioactive sources. Sludge drums are an example of a matrix with a uniform distribution of contaminants. When attempting to perform a gamma assay of a sludge drum, it is very important to adequately simulate this uniform distribution of radionuclides in order to accurately model the assay results. This was accomplished by using a spiral radial source tube placement in a sludge drum rather than the standard three source tubes seen in debris PDP drums. Available line sources (Eu-152) were placed in the spiral tubes to further accomplish the desired uniform distribution of radionuclides. The standard PDP drum (PDP matrix drum 005) and PDP sources were used to determine the lower limits of detection (LLD) and TMU. Analysis results for the sludge drum matrix case for two HPGe detectors were tabulated and evaluated. NNSS has accepted the methodology and results of the measurements towards demonstrating equivalence to CBFO certified systems. In conclusion, the WES-WGS and CMR-OPS gamma spectroscopy teams at LANL have defined and performed measurements that serve to establish and demonstrate equivalency with the processes used by CBFO certified NDA systems. The supplemental measurements address four key areas in Appendix A of DOE/WIPP-02-3122: Annual Calibration Confirmation and Performance Check measurements; LLD determination; and TMU definition. For these measurements the containers, matrices and activity loadings are selected to represent items being assayed in real LLW cases. The LLD and the TMU bounding measurements are to be performed one time and will not be required to be repeated in future campaigns. The annual calibration and performance check measurements were performed initially and planned to repeat in annual campaigns in order to maintain NNSS certification. PDP sources and a PDP sludge drum as well as Eu-152 line sources and a spiral sludge drum were used for the measurements. In all cases, the results for accuracy and precision (%R and %RSD, respectively) were within allowable ranges as defined by the WIPP PDP program. LLD (or MDC) results were established for all the ten WIPP reportable radionuclides and U-235, and the MDC for Pu-239 was established in all cases to be well under 100 nCi/g. Useful results for reducing estimated uncertainties were established and an interesting unexpected case of high bias was observed and will be applied toward this end. (authors)« less

The National Shipbuilding Research Program. Update Handbook for Surface Preparation and Coatings in Tanks and Confined Areas

DTIC Science & Technology

2000-10-31

cleaning method are described in Naval Ships’ Technical Manual Chapter 631. 4.6.4 Citric Acid Cleaning The citric acid cleaning system is intended to...acquisition of necessary chemicals and tools, degreasing/cleaning, paint/stripping/removal, citric acid rust removal, passivation of bare steel, and drying...Figure 9-7 Hanging Explosion -Proof Light Box • Figure 9-8 Lighting in Tank • Figure 10-1 Hazardous Waste Storage Area • Figure 10-2 Solvent

Is Middle-Upper Arm Circumference "normally" distributed? Secondary data analysis of 852 nutrition surveys.

PubMed

Frison, Severine; Checchi, Francesco; Kerac, Marko; Nicholas, Jennifer

2016-01-01

Wasting is a major public health issue throughout the developing world. Out of the 6.9 million estimated deaths among children under five annually, over 800,000 deaths (11.6 %) are attributed to wasting. Wasting is quantified as low Weight-For-Height (WFH) and/or low Mid-Upper Arm Circumference (MUAC) (since 2005). Many statistical procedures are based on the assumption that the data used are normally distributed. Analyses have been conducted on the distribution of WFH but there are no equivalent studies on the distribution of MUAC. This secondary data analysis assesses the normality of the MUAC distributions of 852 nutrition cross-sectional survey datasets of children from 6 to 59 months old and examines different approaches to normalise "non-normal" distributions. The distribution of MUAC showed no departure from a normal distribution in 319 (37.7 %) distributions using the Shapiro-Wilk test. Out of the 533 surveys showing departure from a normal distribution, 183 (34.3 %) were skewed (D'Agostino test) and 196 (36.8 %) had a kurtosis different to the one observed in the normal distribution (Anscombe-Glynn test). Testing for normality can be sensitive to data quality, design effect and sample size. Out of the 533 surveys showing departure from a normal distribution, 294 (55.2 %) showed high digit preference, 164 (30.8 %) had a large design effect, and 204 (38.3 %) a large sample size. Spline and LOESS smoothing techniques were explored and both techniques work well. After Spline smoothing, 56.7 % of the MUAC distributions showing departure from normality were "normalised" and 59.7 % after LOESS. Box-Cox power transformation had similar results on distributions showing departure from normality with 57 % of distributions approximating "normal" after transformation. Applying Box-Cox transformation after Spline or Loess smoothing techniques increased that proportion to 82.4 and 82.7 % respectively. This suggests that statistical approaches relying on the normal distribution assumption can be successfully applied to MUAC. In light of this promising finding, further research is ongoing to evaluate the performance of a normal distribution based approach to estimating the prevalence of wasting using MUAC.

Tuning Riboswitch Regulation through Conformational Selection

PubMed Central

Wilson, Ross C.; Smith, Angela M.; Fuchs, Ryan T.; Kleckner, Ian R.; Henkin, Tina M.; Foster, Mark P.

2010-01-01

SUMMARY The SMK box riboswitch, which represents one of three known classes of S-adenosylmethionine (SAM)-responsive riboswitches, regulates gene expression in bacteria at the level of translation initiation. In contrast to most riboswitches, which contain separate domains responsible for ligand recognition and gene regulation, the ligand-binding and regulatory domains of the SMK box riboswitch are coincident. This property was exploited to allow the first atomic-level characterization of a functionally intact riboswitch in both the ligand-bound and ligand-free states. NMR spectroscopy revealed distinct mutually exclusive RNA conformations that are differentially populated in the presence or absence of the effector metabolite. Isothermal titration calorimetry and in vivo reporter assay results revealed the thermodynamic and functional consequences of this conformational equilibrium. We present a comprehensive model of the structural, thermodynamic, and functional properties of this compact RNA regulatory element. PMID:21075119

[Use of new tools for controlling triatomines in different entomologic situations in the American continent].

PubMed

Oliveira Filho, A M

1997-01-01

The field results of new tools for triatomine control developed under the sponsorship of WHO/TDR are reported: a) A slow-release "paint" containing malathion; b) fumigant cans containing cypermethrin and DDVP, and c) sensor boxes for the detection of triatomine infestations. Field assays were performed in Chile, Honduras and Paraguay against Triatoma infestans, T. dimidiata and Rhodnius prolixus, accordingly to a standard protocol designed by a WHO experts committee. Preliminary 6 months post-treatment results for the three countries show an efficient control when insecticide paints were used indoors and in the peridomicilium, keeping reinfestation near zero. The final results presented for Chile two years post-treatment confirmed the superiority of the slow-release "paints". Sensor boxes were less effective than man/hour captures in the detection of infested houses.

A microfluidic platform with integrated arrays for immunologic assays for biological pathogen detection

NASA Astrophysics Data System (ADS)

Klemm, Richard; Becker, Holger; Hlawatsch, Nadine; Julich, Sandra; Miethe, Peter; Moche, Christian; Schattschneider, Sebastian; Tomaso, Herbert; Gärtner, Claudia

2014-05-01

The ability to integrate complete assays on a microfluidic chip helps to greatly simplify instrument requirements and allows the use of lab-on-a-chip technology in the field. A core application for such field-portable systems is the detection of pathogens in a CBRN scenario such as permanent monitoring of airborne pathogens, e.g. in subway stations or hospitals etc. An immunological assay was chosen as method for the pathogen identification. The conceptual approach was its realization as a lab-on-a-chip system, enabling an easy handling of the sample in an automated manner. The immunological detection takes place on an antibody array directly implemented in the microfluidic network. Different immobilization strategies will be presented showing the performance of the system. Central elements of the disposable microfluidic device like fluidic interface, turning valves, liquid introduction and waste storage, as well as the architecture of measurement and control fluidic network, will be introduced. Overall process times of about 30 minutes were achieved and assays for the detection of Francisella tularensis and Yersinia pestis are presented. An important feature of the integrated lab-on-a-chip approach is that all waste liquids remain on-chip and contamination risks can be avoided.

Evaluation of anaerobic co-digestion of dairy manure with food wastes via bio-methane potential assay and CSTR reactor.

PubMed

Ye, Yulin; Zamalloa, Carlos; Lin, Hongjian; Yan, Mi; Schmidt, David; Hu, Bo

2015-01-01

The introduction of food wastes into anaerobic digestion (AD) brings a promising scenario of increasing feedstock availability and overall energy production from AD. This study evaluated the biodegradability and methane potential from co-digestion of two typical food wastes, kitchen waste and chicken fat, with dairy manure. For single substrate, the bio-methane potential assays showed that kitchen waste had the highest methane yield of 352 L-CH4 kg(-1)-VS added, 92% more than dairy manure alone. Chicken fat at the same Volatile Solid (VS) level (2 g L(-1)) inhibited bio-methane production. Addition of kitchen waste and chicken fat to a VS percentage of up to 40% improved overall methane yield by 44% and 34%, respectively. Synergistic effect was observed when either combining two or three substrates as AD feedstock, possibly as a result of increased biodegradability of organic materials in chicken fat and kitchen waste compared with dairy manure. Addition of chicken fat improved methane yield more than kitchen waste. However, addition of chicken fat VS over 0.8 g L(-1) should be cautiously done because it may cause reactor failure due to decrease in pH. The maximum methane yield was 425 L-CH4 kg(-1)-VS, achieved at a VS ratio of 2:2:1 for kitchen waste, chicken fat, and dairy manure. Results from batch AD experiment demonstrated that supplementing dairy manure to chicken fat and/or kitchen waste improved alkalinity of substrate due to the inclusion of more titratable bases in dairy manure, and therefore stabilized the methanogenesis and substantially improved biogas yield. A mixture of substrates of kitchen waste, chicken fat, and dairy manure at a ratio of 1:1:3 was fed to a continuously stirred tank reactor operated at organic loading rates of 3.28, 6.55, and 2.18 g-COD L(-1)-day (hydraulic retention time of 20, 10, and 30 days, respectively) under mesophilic condition, and methane production rate reached 0.65, 0.95, and 0.34 L-CH4 L(-1)-reactor-day.

Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs.

PubMed

Beyer, Andrea R; VieBrock, Lauren; Rodino, Kyle G; Miller, Daniel P; Tegels, Brittney K; Marconi, Richard T; Carlyon, Jason A

2015-10-01

A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions. Orientia tsutsugamushi, an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here, in silico and manual sequence analyses revealed that a large proportion of O. tsutsugamushi strain Ikeda Anks bear a eukaryotic/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and/or endogenous SCF1 components. When coexpressed with FLAG-Ank4_01 or FLAG-Ank9, a glutathione S-transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated O. tsutsugamushi-derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning O. tsutsugamushi, poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that O. tsutsugamushi strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an O. tsutsugamushi Ank does so during infection, and advance overall understanding of microbial F-box proteins. Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets. Orientia tsutsugamushi, which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that O. tsutsugamushi strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an Orientia-derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide Orientia effector and the first example of an endogenous F-box-containing Ank-mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Orientia tsutsugamushi Strain Ikeda Ankyrin Repeat-Containing Proteins Recruit SCF1 Ubiquitin Ligase Machinery via Poxvirus-Like F-Box Motifs

PubMed Central

Beyer, Andrea R.; VieBrock, Lauren; Rodino, Kyle G.; Miller, Daniel P.; Tegels, Brittney K.; Marconi, Richard T.

2015-01-01

ABSTRACT A rising theme among intracellular microbes is the delivery of ankyrin repeat-containing effectors (Anks) that interact with target proteins to co-opt host cell functions. Orientia tsutsugamushi, an obligate intracellular bacterium and the etiologic agent of scrub typhus, encodes one of the largest Ank repertoires of any sequenced microorganism. They have been previously identified as type 1 secretion system substrates. Here, in silico and manual sequence analyses revealed that a large proportion of O. tsutsugamushi strain Ikeda Anks bear a eukaryotic/poxvirus-like F-box motif, which is known to recruit host cell SCF1 ubiquitin ligase machinery. We assessed the Anks for the ability to serve as F-box proteins. Coimmunoprecipitation assays demonstrated that F-box-containing Anks interact with overexpressed and/or endogenous SCF1 components. When coexpressed with FLAG-Ank4_01 or FLAG-Ank9, a glutathione S-transferase (GST)-tagged version of the SCF1 component SKP1 localized to subcellular sites of FLAG-Ank accumulation. The abilities of recombinant Anks to interact and colocalize with SKP1 were F-box dependent. GST-SKP1 precipitated O. tsutsugamushi-derived Ank9 from infected host cells, verifying both that the pathogen expresses Ank9 during infection and the protein's capability to bind SKP1. Aligning O. tsutsugamushi, poxviral, and eukaryotic F-box sequences delineated three F-box residues that are highly conserved and likely to be functionally important. Substitution of these residues ablated the ability of GFP-Ank9 to interact with GST-SKP1. These results demonstrate that O. tsutsugamushi strain Ikeda Anks can co-opt host cell polyubiquitination machinery, provide the first evidence that an O. tsutsugamushi Ank does so during infection, and advance overall understanding of microbial F-box proteins. IMPORTANCE Ankyrin repeat-containing proteins (Anks) are important virulence factors of intracellular bacteria that mediate protein-protein interactions with host cell targets. Orientia tsutsugamushi, which causes a debilitating infection called scrub typhus in one of the most densely populated regions of the world, encodes one of the largest Ank armamentariums of any sequenced bacterium. This study demonstrates that O. tsutsugamushi strain Ikeda Anks also bear F-box motifs that interact with host cell polyubiquitination machinery. By proving that an Orientia-derived Ank interacts with SKP1 in infected cells, this evidences the first bona fide Orientia effector and the first example of an endogenous F-box-containing Ank–mammalian-host ligand interaction for any intracellular bacterium. Also, importantly, this work identifies key residues that are essential for microbial F-box function. PMID:26170417

Heterologous Expression and Molecular and Cellular Characterization of CaPUB1 Encoding a Hot Pepper U-Box E3 Ubiquitin Ligase Homolog1[C

PubMed Central

Cho, Seok Keun; Chung, Hoo Sun; Ryu, Moon Young; Park, Mi Jin; Lee, Myeong Min; Bahk, Young-Yil; Kim, Jungmook; Pai, Hyun Sook; Kim, Woo Taek

2006-01-01

The U-box motif is a conserved domain found in the diverse isoforms of E3 ubiquitin ligase in eukaryotes. From water-stressed hot pepper (Capsicum annuum L. cv Pukang) plants, we isolated C. annuum putative U-box protein 1 (CaPUB1), which encodes a protein containing a single U-box motif in its N-terminal region. In vitro ubiquitination and site-directed mutagenesis assays revealed that CaPUB1 possessed E3 ubiquitin ligase activity and that the U-box motif was indeed essential for its enzyme activity. RNA gel-blot analysis showed that CaPUB1 mRNA was induced rapidly by a broad spectrum of abiotic stresses, including drought, high salinity, cold temperature, and mechanical wounding, but not in response to ethylene, abscisic acid, or a bacterial pathogen, suggesting its role in the early events in the abiotic-related defense response. Because transgenic work was extremely difficult in hot pepper, in this study we overexpressed CaPUB1 in Arabidopsis (Arabidopsis thaliana) to provide cellular information on the function of this gene in the development and plant responses to abiotic stresses. Transgenic Arabidopsis plants that constitutively expressed the CaPUB1 gene under the control of the cauliflower mosaic virus 35S promoter had markedly longer hypocotyls and roots and grew more rapidly than the wild type, leading to an early bolting phenotype. Microscopic analysis showed that 35S∷CaPUB1 roots had increased numbers of small-sized cells, resulting in disordered, highly populated cell layers in the cortex, endodermis, and stele. In addition, CaPUB1-overexpressing plants displayed increased sensitivity to water stress and mild salinity. These results indicate that CaPUB1 is functional in Arabidopsis cells, thereby effectively altering cell and tissue growth and also the response to abiotic stresses. Comparative proteomic analysis showed that the level of RPN6 protein, a non-ATPase subunit of the 26S proteasome complex, was significantly reduced in 35S∷CaPUB1 seedlings as compared to the wild type. Pull-down and ubiquitination assays demonstrated that RPN6 interacted physically with CaPUB1 and was ubiquitinated in a CaPUB1-dependent manner in vitro. Although the physiological function of CaPUB1 is not yet clear, there are several possibilities for its involvement in a subset of physiological responses to counteract dehydration and high-salinity stresses in transgenic Arabidopsis seedlings. PMID:17041029

Estrogen Maintains Skeletal Muscle in Septic Rats Associated with Altering Hypothalamic Inflammation and Neuropeptides.

PubMed

Zhao, Chenyan; Li, Jun; Cheng, Minhua; Shi, Jialing; Shen, Juanhong; Gao, Tao; Xi, Fengchan; Yu, Wenkui

2017-03-01

Muscle wasting is one of the main contributors to the worse outcomes in sepsis. Whether estrogen could alleviate muscle wasting induced by sepsis remains unclear. This study was designed to test the effect of estrogen on muscle wasting and its relationship with central alteration in sepsis. Thirty Sprague-Dawley rats were divided into 3 groups: control group, sepsis group, and estrogen treated sepsis group. Animals were intraperitoneally injected with lipopolysaccharide (10 mg/kg) or saline, followed by subcutaneous injection of 17β-estradiol (1 mg/kg) or saline. Twenty-four hours later, all animals were killed and their hypothalamus and skeletal muscles were harvested for analysis. Muscle wasting markers, hypothalamic neuropeptides, and hypothalamic inflammatory markers were measured. As a result, lipopolysaccharide administration caused a significant increase in muscle wasting, hypothalamic inflammation, and anorexigenic neuropeptides (POMC and CART) gene expression, and a significant decrease in orexigenic neuropeptides (AgRP and NPY) gene expression. Administration of estrogen signifcantl attenuated lipopolysaccharide-induced muscle wasting (body weight and extensor digitorum longus loss [52 and 62 %], tyrosine and 3-methylhistidine release [17 and 22 %], muscle ring fnger 1 [MuRF-1; 65 %], and muscle atrophy F-box [MAFbx] gene expression), hypothalamic inflammation (Tumor necrosis factor-α and interlukin-1β [69 and 70%]) as well as alteration of POMC, CART and AgRP (61, 37, and 1008 %) expression.In conclusion, estrogen could alleviate sepsis-induced muscle wasting and it was associated with reducing hypothalamic inflammation and alteration of hypothalamic neuropeptides. © Georg Thieme Verlag KG Stuttgart · New York.

Light directs zebrafish period2 expression via conserved D and E boxes.

PubMed

Vatine, Gad; Vallone, Daniela; Appelbaum, Lior; Mracek, Philipp; Ben-Moshe, Zohar; Lahiri, Kajori; Gothilf, Yoav; Foulkes, Nicholas S

2009-10-01

For most species, light represents the principal environmental signal for entraining the endogenous circadian clock. The zebrafish is a fascinating vertebrate model for studying this process since unlike mammals, direct exposure of most of its tissues to light leads to local clock entrainment. Importantly, light induces the expression of a set of genes including certain clock genes in most zebrafish cell types in vivo and in vitro. However, the mechanism linking light to gene expression remains poorly understood. To elucidate this key mechanism, here we focus on how light regulates transcription of the zebrafish period2 (per2) gene. Using transgenic fish and stably transfected cell line-based assays, we define a Light Responsive Module (LRM) within the per2 promoter. The LRM lies proximal to the transcription start site and is both necessary and sufficient for light-driven gene expression and also for a light-dependent circadian clock regulation. Curiously, the LRM sequence is strongly conserved in other vertebrate per2 genes, even in species lacking directly light-sensitive peripheral clocks. Furthermore, we reveal that the human LRM can substitute for the zebrafish LRM to confer light-regulated transcription in zebrafish cells. The LRM contains E- and D-box elements that are critical for its function. While the E-box directs circadian clock regulation by mediating BMAL/CLOCK activity, the D-box confers light-driven expression. The zebrafish homolog of the thyrotroph embryonic factor binds efficiently to the LRM D-box and transactivates expression. We demonstrate that tef mRNA levels are light inducible and that knock-down of tef expression attenuates light-driven transcription from the per2 promoter in vivo. Together, our results support a model where a light-dependent crosstalk between E- and D-box binding factors is a central determinant of per2 expression. These findings extend the general understanding of the mechanism whereby the clock is entrained by light and how the regulation of clock gene expression by light has evolved in vertebrates.

Effects of substrate to inoculum ratio on the biochemical methane potential of piggery slaughterhouse wastes.

PubMed

Yoon, Young-Man; Kim, Seung-Hwan; Shin, Kook-Sik; Kim, Chang-Hyun

2014-04-01

The aim of this study was to assess the effect of substrate to inoculum ratio (S/I ratio) on the biochemical methane potential (BMP) and anaerobic biodegradability (Ddeg) of different piggery slaughterhouse wastes, such as piggery blood, intestine residue, and digestive tract content. These wastes were sampled from a piggery slaughterhouse located in Kimje, South Korea. Cumulative methane production curves for the wastes were obtained from the anaerobic batch fermentation having different S/I ratios of 0.1, 0.5, 1.0, and 1.5. BMP and anaerobic biodegradabilities (Ddeg) of the wastes were calculated from cumulative methane production data for the tested conditions. At the lowest S/I ration of 0.1, BMPs of piggery blood, intestine residue, and digestive tract content were determined to be 0.799, 0.848, and 1.076 Nm(3) kg(-1)-VSadded, respectively, which were above the theoretical methane potentials of 0.539, 0.644, and 0.517 Nm(3) kg(-1)-VSadded for blood, intestine residue, and digestive tract content, respectively. However, BMPs obtained from the higher S/I ratios of 0.5, 1.0, and 1.5 were within the theoretical range for all three types of waste and were not significantly different for the different S/I ratios tested. Anaerobic biodegradabilities calculated from BMP data showed a similar tendency. These results imply that, for BMP assay in an anaerobic reactor, the S/I ratio of anaerobic reactor should be above 0.1 and the inoculum should be sufficiently stabilized to avoid further degradation during the assay.

Effects of Substrate to Inoculum Ratio on the Biochemical Methane Potential of Piggery Slaughterhouse Wastes

PubMed Central

Yoon, Young-Man; Kim, Seung-Hwan; Shin, Kook-Sik; Kim, Chang-Hyun

2014-01-01

The aim of this study was to assess the effect of substrate to inoculum ratio (S/I ratio) on the biochemical methane potential (BMP) and anaerobic biodegradability (Ddeg) of different piggery slaughterhouse wastes, such as piggery blood, intestine residue, and digestive tract content. These wastes were sampled from a piggery slaughterhouse located in Kimje, South Korea. Cumulative methane production curves for the wastes were obtained from the anaerobic batch fermentation having different S/I ratios of 0.1, 0.5, 1.0, and 1.5. BMP and anaerobic biodegradabilities (Ddeg) of the wastes were calculated from cumulative methane production data for the tested conditions. At the lowest S/I ration of 0.1, BMPs of piggery blood, intestine residue, and digestive tract content were determined to be 0.799, 0.848, and 1.076 Nm3 kg−1-VSadded, respectively, which were above the theoretical methane potentials of 0.539, 0.644, and 0.517 Nm3 kg−1-VSadded for blood, intestine residue, and digestive tract content, respectively. However, BMPs obtained from the higher S/I ratios of 0.5, 1.0, and 1.5 were within the theoretical range for all three types of waste and were not significantly different for the different S/I ratios tested. Anaerobic biodegradabilities calculated from BMP data showed a similar tendency. These results imply that, for BMP assay in an anaerobic reactor, the S/I ratio of anaerobic reactor should be above 0.1 and the inoculum should be sufficiently stabilized to avoid further degradation during the assay. PMID:25049994

OsBIRH1, a DEAD-box RNA helicase with functions in modulating defence responses against pathogen infection and oxidative stress

PubMed Central

Li, Dayong; Liu, Huizhi; Zhang, Huijuan; Wang, Xiaoe; Song, Fengming

2008-01-01

DEAD-box proteins comprise a large protein family with members from all kingdoms and play important roles in all types of processes in RNA metabolism. In this study, a rice gene OsBIRH1, which encodes a DEAD-box RNA helicase protein, was cloned and characterized. The predicted OsBIRH1 protein contains a DEAD domain and all conserved motifs that are common characteristics of DEAD-box RNA helicases. Recombinant OsBIRH1 protein purified from Escherichia coli was shown to have both RNA-dependent ATPase and ATP-dependent RNA helicase activities in vitro. Expression of OsBIRH1 was activated in rice seedling leaves after treatment with defence-related signal chemicals, for example benzothiadiazole, salicylic acid, l-aminocyclopropane-1-carboxylic acid, and jasmonic acid, and was also up-regulated in an incompatible interaction between a resistant rice genotype and the blast fungus, Magnaporthe grisea. Transgenic Arabidopsis plants that overexpress the OsBIRH1 gene were generated. Disease resistance phenotype assays revealed that the OsBIRH1-overexpressing transgenic plants showed an enhanced disease resistance against Alternaria brassicicola and Pseudomonas syringae pv. tomato DC3000. Meanwhile, defence-related genes, for example PR-1, PR-2, PR-5, and PDF1.2, showed an up-regulated expression in the transgenic plants. Moreover, the OsBIRH1 transgenic Arabidopsis plants also showed increased tolerance to oxidative stress and elevated expression levels of oxidative defence genes, AtApx1, AtApx2, and AtFSD1. The results suggest that OsBIRH1 encodes a functional DEAD-box RNA helicase and plays important roles in defence responses against biotic and abiotic stresses. PMID:18441339

Differential transcriptional regulation of banana sucrose phosphate synthase gene in response to ethylene, auxin, wounding, low temperature and different photoperiods during fruit ripening and functional analysis of banana SPS gene promoter.

PubMed

Roy Choudhury, Swarup; Roy, Sujit; Das, Ranjan; Sengupta, Dibyendu N

2008-12-01

Sucrose phosphate synthase (SPS) (EC 2.3.1.14) is the key regulatory component in sucrose formation in banana (Musa acuminata subgroup Cavendish, cv Giant governor) fruit during ripening. This report illustrates differential transcriptional responses of banana SPS gene following ethylene, auxin, wounding, low temperature and different photoperiods during ripening in banana fruit. Whereas ethylene strongly stimulated SPS transcript accumulation, auxin and cold treatment only marginally increased the abundance of SPS mRNA level, while wounding negatively regulated SPS gene expression. Conversely, SPS transcript level was distinctly increased by constant exposure to white light. Protein level, enzymatic activity of SPS and sucrose synthesis were substantially increased by ethylene and increased exposure to white light conditions as compared to other treatments. To further study the transcriptional regulation of SPS in banana fruit, the promoter region of SPS gene was cloned and some cis-acting regulatory elements such as a reverse GCC-box ERE, two ARE motifs (TGTCTC), one LTRE (CCGAA), a GAGA-box (GAGA...) and a GATA-box LRE (GATAAG) were identified along with the TATA and CAAT-box. DNA-protein interaction studies using these cis-elements indicated a highly specific cis-trans interaction in the banana nuclear extract. Furthermore, we specifically studied the light responsive characteristics of GATA-box containing synthetic as well as native banana SPS promoter. Transient expression assays using banana SPS promoter have also indicated the functional importance of the SPS promoter in regulating gene expression. Together, these results provide insights into the transcriptional regulation of banana SPS gene in response to phytohormones and other environmental factors during fruit ripening.

What level of estrogenic activity determined by in vitro assays in municipal waste waters can be considered as safe?

PubMed

Jarošová, Barbora; Bláha, Luděk; Giesy, John P; Hilscherová, Klára

2014-03-01

In vitro assays are broadly used tools to evaluate the estrogenic activity in Waste Water Treatment Plant (WWTP) effluents and their receiving rivers. Since potencies of individual estrogens to induce in vitro and in vivo responses can differ it is not possible to directly evaluate risks based on in vitro measures of estrogenic activity. Estrone, 17beta-estradiol, 17alfa-ethinylestradiol and to some extent, estriol have been shown to be responsible for the majority of in vitro estrogenic activity of municipal WWTP effluents. Therefore, in the present study safe concentrations of Estrogenic Equivalents (EEQs-SSE) in municipal WWTP effluents were derived based on simplified assumption that the steroid estrogens are responsible for all estrogenicity determined with particular in vitro assays. EEQs-SSEs were derived using the bioassay and testing protocol-specific in vitro potencies of steroid estrogens, in vivo predicted no effect concentration (PNECs) of these compounds, and their relative contributions to the overall estrogenicity detected in municipal WWTP effluents. EEQs-SSEs for 15 individual bioassays varied from 0.1 to 0.4ng EEQ/L. The EEQs-SSEs are supposed to be increased by use of location-specific dilution factors of WWTP effluents entering receiving rivers. They are applicable to municipal wastewater and rivers close to their discharges, but not to industrial waste waters. Copyright © 2013 Elsevier Ltd. All rights reserved.

Canine scent detection and microbial source tracking of human waste contamination in storm drains.

PubMed

Van De Werfhorst, Laurie C; Murray, Jill L S; Reynolds, Scott; Reynolds, Karen; Holden, Patricia A

2014-06-01

Human fecal contamination of surface waters and drains is difficult to diagnose. DNA-based and chemical analyses of water samples can be used to specifically quantify human waste contamination, but their expense precludes routine use. We evaluated canine scent tracking, using two dogs trained to respond to the scent of municipal wastewater, as a field approach for surveying human fecal contamination. Fecal indicator bacteria, as well as DNA-based and chemical markers of human waste, were analyzed in waters sampled from canine scent-evaluated sites (urban storm drains and creeks). In the field, the dogs responded positively (70% and 100%) at sites for which sampled waters were then confirmed as contaminated with human waste. When both dogs indicated a negative response, human waste markers were absent. Overall, canine scent tracking appears useful for prioritizing sampling sites for which DNA-based and similarly expensive assays can confirm and quantify human waste contamination.

A simple and predictive phenotypic High Content Imaging assay for Plasmodium falciparum mature gametocytes to identify malaria transmission blocking compounds

PubMed Central

Lucantoni, Leonardo; Silvestrini, Francesco; Signore, Michele; Siciliano, Giulia; Eldering, Maarten; Dechering, Koen J.; Avery, Vicky M.; Alano, Pietro

2015-01-01

Plasmodium falciparum gametocytes, specifically the mature stages, are the only malaria parasite stage in humans transmissible to the mosquito vector. Anti-malarial drugs capable of killing these forms are considered essential for the eradication of malaria and tools allowing the screening of large compound libraries with high predictive power are needed to identify new candidates. As gametocytes are not a replicative stage it is difficult to apply the same drug screening methods used for asexual stages. Here we propose an assay, based on high content imaging, combining “classic” gametocyte viability readout based on gametocyte counts with a functional viability readout, based on gametocyte activation and the discrimination of the typical gamete spherical morphology. This simple and rapid assay has been miniaturized to a 384-well format using acridine orange staining of wild type P. falciparum 3D7A sexual forms, and was validated by screening reference antimalarial drugs and the MMV Malaria Box. The assay demonstrated excellent robustness and ability to identify quality hits with high likelihood of confirmation of transmission reducing activity in subsequent mosquito membrane feeding assays. PMID:26553647

Review and Implementation of Technology for Solid Radioactive Waste Volume Reduction

DTIC Science & Technology

1999-10-15

were shifted to Project 1.1 for spent nuclear fuel cask development to accelerate that project. Those funds should be repaid to Project 1.3 in the... transported between the shipyards such as Nerpa, and other intermediate storage sites such as Gremikha and Andreeva Bay. At these sites the largest...waste source and allow pretreatment unit operations using commercially available technologies of contaminant assaying, cutting/shearing, sorting

Segmented Gamma Scanner for Small Containers of Uranium Processing Waste- 12295

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morris, K.E.; Smith, S.K.; Gailey, S.

2012-07-01

The Segmented Gamma Scanner (SGS) is commonly utilized in the assay of 55-gallon drums containing radioactive waste. Successfully deployed calibration methods include measurement of vertical line source standards in representative matrices and mathematical efficiency calibrations. The SGS technique can also be utilized to assay smaller containers, such as those used for criticality safety in uranium processing facilities. For such an application, a Can SGS System is aptly suited for the identification and quantification of radionuclides present in fuel processing wastes. Additionally, since the significant presence of uranium lumping can confound even a simple 'pass/fail' measurement regimen, the high-resolution gamma spectroscopymore » allows for the use of lump-detection techniques. In this application a lump correction is not required, but the application of a differential peak approach is used to simply identify the presence of U-235 lumps. The Can SGS is similar to current drum SGSs, but differs in the methodology for vertical segmentation. In the current drum SGS, the drum is placed on a rotator at a fixed vertical position while the detector, collimator, and transmission source are moved vertically to effect vertical segmentation. For the Can SGS, segmentation is more efficiently done by raising and lowering the rotator platform upon which the small container is positioned. This also reduces the complexity of the system mechanism. The application of the Can SGS introduces new challenges to traditional calibration and verification approaches. In this paper, we revisit SGS calibration methodology in the context of smaller waste containers, and as applied to fuel processing wastes. Specifically, we discuss solutions to the challenges introduced by requiring source standards to fit within the confines of the small containers and the unavailability of high-enriched uranium source standards. We also discuss the implementation of a previously used technique for identifying the presence of uranium lumping. The SGS technique is a well-accepted NDA technique applicable to containers of almost any size. It assumes a homogenous matrix and activity distribution throughout the entire container; an assumption that is at odds with the detection of lumps within the assay item typical of uranium-processing waste. This fact, in addition to the difficultly in constructing small reference standards of uranium-bearing materials, required the methodology used for performing an efficiency curve calibration to be altered. The solution discussed in this paper is demonstrated to provide good results for both the segment activity and full container activity when measuring heterogeneous source distributions. The application of this approach will need to be based on process knowledge of the assay items, as biases can be introduced if used with homogenous, or nearly homogenous, activity distributions. The bias will need to be quantified for each combination of container geometry and SGS scanning settings. One recommended approach for using the heterogeneous calibration discussed here is to assay each item using a homogenous calibration initially. Review of the segment activities compared to the full container activity will signal the presence of a non-uniform activity distribution as the segment activity will be grossly disproportionate to the full container activity. Upon seeing this result, the assay should either be reanalyzed or repeated using the heterogeneous calibration. (authors)« less

Comet assay and micronucleus tests on Oreochromis niloticus (Perciforme: Cichlidae) exposed to raw sugarcane vinasse and to phisicochemical treated vinasse by pH adjustment with lime (CaO).

PubMed

Correia, Jorge E; Christofoletti, Cintya Ap; Ansoar-Rodríguez, Yadira; Guedes, Thays A; Fontanetti, Carmem S

2017-04-01

In Brazil vinasse, a main sugarcane distillery residue, stands out because every liter of alcohol generates 10-15 L of vinasse as waste. An alternative for the disposal of this waste is the fertirrigation of the sugarcane culture itself. However, the high amount released can saturate the soil and through leaching/percolation contaminate water resources. The aim of this study is verifying the toxic potential of vinasse in tilapias and effectiveness of the physicalchemical treatment of this waste with pH adjustment with lime (CaO). The comet assay and the micronucleus test were applied on animals exposed to dilutions of raw vinasse and vinasse adjusted to neutral pH. Bioassays with raw vinasse dilutions indicated a toxic and genotoxic potential; fish exposed to the highest concentration died less than 48 h after the exposure; the incidence of micronucleus was significantly higher when compared to negative control for all dilutions. For the comet assay, the scores of damage were statistically higher for all dilutions, with the exception of the 1% dillution. However, in the bioassay with the chemically treated vinasse (neutral pH), most fish in the 10% dilution survived and there was no significant difference when compared to the control. Damage scores in the comet assay were similar to the results of the untreated vinasse. The chemical treatment of vinasse with lime to neutralize the pH proved to be an effective alternative for the toxicity reduction of this residue, since it reduced the mortality of fish at higher concentrations and the incidence of damage to DNA. Copyright © 2017 Elsevier Ltd. All rights reserved.

Application of Response Surface Methodology on Leaching of Iron from Partially Laterised Khondalite Rocks: A Bauxite Mining Waste

NASA Astrophysics Data System (ADS)

Swain, Ranjita; Bhima Rao, R.

2018-04-01

In the present investigation, response surface methodology (RSM) is used for a quadratic model that continuously controls the process parameters. This model is used to optimize the removal of iron oxide from Partially Laterised Khondalite (PLK) rocks which is influenced by several independent variables namely acid concentration, time and temperature. Second order response functions are produced for leaching of iron oxide from PLK rocks-a bauxite mining waste. In RSM, Box-Behnken design is used for the process optimization to achieve maximum removal of iron oxide. The influence of the process variables of leaching of iron oxide is presented in the form of 3-D response graphs. The results of this investigation reveals that 3 M hydrochloric acid concentration, 240 min time and 373 K temperature are found to be the best conditions for removal of 99% Fe2O3. The product obtain at this condition contain 80% brightness which is suitable for ceramic and filler industry applications. The novelity of the work is that the waste can be a value added product after suitable physical beneficiation and chemical treatment.

Prototype Development of Remote Operated Hot Uniaxial Press (ROHUP) to Fabricate Advanced Tc-99 Bearing Ceramic Waste Forms - 13381

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alaniz, Ariana J.; Delgado, Luc R.; Werbick, Brett M.

The objective of this senior student project is to design and build a prototype construction of a machine that simultaneously provides the proper pressure and temperature parameters to sinter ceramic powders in-situ to create pellets of rather high densities of above 90% (theoretical). This ROHUP (Remote Operated Hot Uniaxial Press) device is designed specifically to fabricate advanced ceramic Tc-99 bearing waste forms and therefore radiological barriers have been included in the system. The HUP features electronic control and feedback systems to set and monitor pressure, load, and temperature parameters. This device operates wirelessly via portable computer using Bluetooth{sup R} technology.more » The HUP device is designed to fit in a standard atmosphere controlled glove box to further allow sintering under inert conditions (e.g. under Ar, He, N{sub 2}). This will further allow utilizing this HUP for other potential applications, including radioactive samples, novel ceramic waste forms, advanced oxide fuels, air-sensitive samples, metallic systems, advanced powder metallurgy, diffusion experiments and more. (authors)« less

Improving the 'tool box' for robust industrial enzymes.

PubMed

Littlechild, J A

2017-05-01

The speed of sequencing of microbial genomes and metagenomes is providing an ever increasing resource for the identification of new robust biocatalysts with industrial applications for many different aspects of industrial biotechnology. Using 'natures catalysts' provides a sustainable approach to chemical synthesis of fine chemicals, general chemicals such as surfactants and new consumer-based materials such as biodegradable plastics. This provides a sustainable and 'green chemistry' route to chemical synthesis which generates no toxic waste and is environmentally friendly. In addition, enzymes can play important roles in other applications such as carbon dioxide capture, breakdown of food and other waste streams to provide a route to the concept of a 'circular economy' where nothing is wasted. The use of improved bioinformatic approaches and the development of new rapid enzyme activity screening methodology can provide an endless resource for new robust industrial biocatalysts.This mini-review will discuss several recent case studies where industrial enzymes of 'high priority' have been identified and characterised. It will highlight specific hydrolase enzymes and recent case studies which have been carried out within our group in Exeter.

Criticality assessment of LLRWDF closure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sarrack, A.G.; Weber, J.H.; Woody, N.D.

1992-10-06

During the operation of the Low Level Radioactive Waste Disposal Facility (LLRWDF), large amounts (greater than 100 kg) of enriched uranium (EU) were buried. This EU came primarily from the closing and decontamination of the Naval Fuels Facility in the time period from 1987--1989. Waste Management Operations (WMO) procedures were used to keep the EU boxes separated to prevent possible criticality during normal operation. Closure of the LLRWDF is currently being planned, and waste stabilization by Dynamic Compaction (DC) is proposed. Dynamic compaction will crush the containers in the LLRWDF and result in changes in their geometry. Research of themore » LLRWDF operations and record keeping practices have shown that the EU contents of trenches are known, but details of the arrangement of the contents cannot be proven. Reviews of the trench contents, combined with analysis of potential critical configurations, revealed that some portions of the LLRWDF can be expected to be free of criticality concerns while other sections have credible probabilities for the assembly of a critical mass, even in the uncompacted configuration. This will have an impact on the closure options and which trenches can be compacted.« less

Dormancy-Associated MADS-Box (DAM) and the Abscisic Acid Pathway Regulate Pear Endodormancy Through a Feedback Mechanism.

PubMed

Tuan, Pham Anh; Bai, Songling; Saito, Takanori; Ito, Akiko; Moriguchi, Takaya

2017-08-01

In the pear 'Kosui' (Pyrus pyrifolia Nakai), the dormancy-associated MADS-box (PpDAM1 = PpMADS13-1) gene has been reported to play an essential role in bud endodormancy. Here, we found that PpDAM1 up-regulated expression of 9-cis-epoxycarotenoid dioxygenase (PpNCED3), which is a rate-limiting gene for ABA biosynthesis. Transient assays with a dual luciferase reporter system (LUC assay) and electrophoretic mobility shift assay (EMSA) showed that PpDAM1 activated PpNCED3 expression by binding to the CArG motif in the PpNCED3 promoter. PpNCED3 expression was increased toward endodormancy release in lateral flower buds of 'Kosui', which is consistent with the induced levels of ABA, its catabolism (ABA 8'-hydroxylase) and signaling genes (type 2C protein phosphatase genes and SNF1-related protein kinase 2 genes). In addition, we found that an ABA response element (ABRE)-binding transcription factor, PpAREB1, exhibiting high expression concomitant with endodormancy release, bound to three ABRE motifs in the promoter region of PpDAM1 and negatively regulated its activity. Taken together, our results suggested a feedback regulation between PpDAM1 and the ABA metabolism and signaling pathway during endodormancy of pear. This first evidence of an interaction between a DAM and ABA biosynthesis in vitro will provide further insights into bud endodormancy regulatory mechanisms of deciduous trees including pear. © The Author 2017. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Identification of Four-Jointed Box 1 (FJX1)-Specific Peptides for Immunotherapy of Nasopharyngeal Carcinoma

PubMed Central

Chai, San Jiun; Yap, Yoke Yeow; Foo, Yoke Ching; Yap, Lee Fah; Ponniah, Sathibalan; Teo, Soo Hwang; Cheong, Sok Ching; Patel, Vyomesh; Lim, Kue Peng

2015-01-01

Nasopharyngeal carcinoma (NPC) is highly prevalent in South East Asia and China. The poor outcome is due to late presentation, recurrence, distant metastasis and limited therapeutic options. For improved treatment outcome, immunotherapeutic approaches focusing on dendritic and autologous cytotoxic T-cell based therapies have been developed, but cost and infrastructure remain barriers for implementing these in low-resource settings. As our prior observations had found that four-jointed box 1 (FJX1), a tumor antigen, is overexpressed in NPCs, we investigated if short 9–20 amino acid sequence specific peptides matching to FJX1 requiring only intramuscular immunization to train host immune systems would be a better treatment option for this disease. Thus, we designed 8 FJX1-specific peptides and implemented an assay system to first, assess the binding of these peptides to HLA-A2 molecules on T2 cells. After, ELISPOT assays were used to determine the peptides immunogenicity and ability to induce potential cytotoxicity activity towards cancer cells. Also, T-cell proliferation assay was used to evaluate the potential of MHC class II peptides to stimulate the expansion of isolated T-cells. Our results demonstrate that these peptides are immunogenic and peptide stimulated T-cells were able to induce peptide-specific cytolytic activity specifically against FJX1-expressing cancer cells. In addition, we demonstrated that the MHC class II peptides were capable of inducing T-cell proliferation. Our results suggest that these peptides are capable of inducing specific cytotoxic cytokines secretion against FJX1-expressing cancer cells and serve as a potential vaccine-based therapy for NPC patients. PMID:26536470

The tight junction protein ZO-1 and an interacting transcription factor regulate ErbB-2 expression

PubMed Central

Balda, Maria S.; Matter, Karl

2000-01-01

Epithelial tight junctions regulate paracellular diffusion and restrict the intermixing of apical and basolateral plasma membrane components. We now identify a Y-box transcription factor, ZONAB (ZO-1-associated nucleic acid-binding protein), that binds to the SH3 domain of ZO-1, a submembrane protein of tight junctions. ZONAB localizes to the nucleus and at tight junctions, and binds to sequences of specific promoters containing an inverted CCAAT box. In reporter assays, ZONAB and ZO-1 functionally interact in the regulation of the ErbB-2 promoter in a cell density-dependent manner. In stably transfected overexpressing cells, ZO-1 and ZONAB control expression of endogenous ErbB-2 and function in the regulation of paracellular permeability. These data indicate that tight junctions directly participate in the control of gene expression and suggest that they function in the regulation of epithelial cell differentiation. PMID:10790369

Formulation and optimization of oxaliplatin immuno-nanoparticles using Box-Behnken design and cytotoxicity assessment for synergistic and receptor-mediated targeting in the treatment of colorectal cancer.

PubMed

Tummala, Shashank; Gowthamarajan, K; Satish Kumar, M N; Praveen, T K; Yamjala, Karthik; Tripuraneni, Naga Srinivas; Prakash, Ashwati

2016-12-01

Conventional chemotherapy majorly lacks clinical application attributed to its inspecificity, adverse effects and inability to penetrate into tumor cells. Hence, the aim of the study was to prepare oxaliplatin solid lipid nanoparticles (OP-SLN) by microemulsion method optimizing it by Box-Behnken design and then covalently conjugated to TRAIL (CD-253) monoclonal antibody (TR-OP-SLN) for targeting colorectal cancer cells. The optimized OP-SLN3 has shown an appreciable particle size (121 ± 1.22 nm), entrapment efficiency (78 ± 0.09%) and drug loading (32 ± 1.01%). Fluorescence study and the Bradford assay further confirmed the binding of the protein. A 1.5-fold increase in cytotoxicity of immuno-nanoparticles (4.9 μM) was observed.

Closure Report for Corrective Action Unit 516: Septic Systems and Discharge Points

DOE Office of Scientific and Technical Information (OSTI.GOV)

NSTec Environmental Restoration

Corrective Action Unit (CAU) 516 is located in Areas 3, 6, and 22 of the Nevada Test Site. CAU 516 is listed in the Federal Facility Agreement and Consent Order of 1996 as Septic Systems and Discharge Points, and is comprised of six Corrective Action Sites (CASs): {sm_bullet} CAS 03-59-01, Bldg 3C-36 Septic System {sm_bullet} CAS 03-59-02, Bldg 3C-45 Septic System {sm_bullet} CAS 06-51-01, Sump and Piping {sm_bullet} CAS 06-51-02, Clay Pipe and Debris {sm_bullet} CAS 06-51-03, Clean Out Box and Piping {sm_bullet} CAS 22-19-04, Vehicle Decontamination Area The Nevada Division of Environmental Protection (NDEP)-approved corrective action alternative for CASsmore » 06-51-02 and 22-19-04 is no further action. The NDEP-approved corrective action alternative for CASs 03-59-01, 03-59-02, 06-51-01, and 06-51-03 is clean closure. Closure activities included removing and disposing of total petroleum hydrocarbon (TPH)-impacted septic tank contents, septic tanks, distribution/clean out boxes, and piping. CAU 516 was closed in accordance with the NDEP-approved CAU 516 Corrective Action Plan (CAP). The closure activities specified in the CAP were based on the recommendations presented in the CAU 516 Corrective Action Decision Document (U.S. Department of Energy, National Nuclear Security Administration Nevada Site Office, 2004). This Closure Report documents CAU 516 closure activities. During closure activities, approximately 186 tons of hydrocarbon waste in the form of TPH-impacted soil and debris, as well as 89 tons of construction debris, were generated and managed and disposed of appropriately. Waste minimization techniques, such as field screening of soil samples and the utilization of laboratory analysis to characterize and classify waste streams, were employed during the performance of closure work.« less

Potential of utilizing asphalt dust waste as filler material in the production of sustainable self compacting concrete (SCC)

NASA Astrophysics Data System (ADS)

Ismail, Isham; Shahidan, Shahiron; Bahari, Nur Amira Afiza Saiful

2017-12-01

Waste materials from many industries are widely used in the production of sustainable green concrete. Utilizing asphalt dust waste (ADW) as a filler material in the development of self-compacting concrete (SCC) is one of the alternative solutions for reducing environmental waste. SCC is an innovative concrete that does not require vibration for placing and compaction. However, there is limited information on the effects of utilizing ADW in the development of SCC. Therefore, this research study examines the effects of various w/b ratios (0.2, 0.3 and 0.4) and differing amounts of ADW (0% to 50%) on the rheological properties of fresh state concrete. The compressive strength of the SCC was tested only for 7 and 28 days as preliminary studies. The results revealed that mixtures MD730, MD740 and MD750 showed satisfactory results for the slump flow, J-Ring, L-Box and V-Funnel test during the fresh state. The compressive strength values obtained after 28 days for MD730, MD740 and MD750 were 35.1 MPa, 36.8 MPa and 29.4 MPa respectively. In conclusion, the distribution of materials in mixtures has significant effect in achieving rheological properties and compressive strength of SCC.

Bioengineering thermodynamics of biological cells.

PubMed

Lucia, Umberto

2015-12-01

Cells are open complex thermodynamic systems. They can be also regarded as complex engines that execute a series of chemical reactions. Energy transformations, thermo-electro-chemical processes and transports phenomena can occur across the cells membranes. Moreover, cells can also actively modify their behaviours in relation to changes in their environment. Different thermo-electro-biochemical behaviours occur between health and disease states. But, all the living systems waste heat, which is no more than the result of their internal irreversibility. This heat is dissipated into the environment. But, this wasted heat represent also a sort of information, which outflows from the cell toward its environment, completely accessible to any observer. The analysis of irreversibility related to this wasted heat can represent a new approach to study the behaviour of the cells themselves and to control their behaviours. So, this approach allows us to consider the living systems as black boxes and analyze only the inflows and outflows and their changes in relation to the modification of the environment. Therefore, information on the systems can be obtained by analyzing the changes in the cell heat wasted in relation to external perturbations. The bioengineering thermodynamics bases are summarized and used to analyse possible controls of the calls behaviours based on the control of the ions fluxes across the cells membranes.

Correlative analysis of heavy metal bioconcentration and genetic damage in white-footed mice (Peromyscus leucopus) from a hazardous waste site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tull-Singleton, S.; Kimball, S.; McBee, K.

1994-05-01

Heavy metals are common constituents of hazardous waste sites and may cause health problems in wildlife and nearby human residents. Previous studies have been conducted on the bioaccumulation/bioconcentration of metals in biological tissue using small mammals. However, there have been few attempts to correlate tissue residues with other physiological or genetic biomarkers. In this study, livers of Peromyscus leucopus (white footed mouse) from a hazardous waste site and a matched reference site were analyzed for the presence of selected metals. The white-footed mouse prefers wooded, brushy habitat. The diet included seeds, nuts, plant material, fungi, and some invertebrates. Waste sitemore » animals were obtained from a facility in southcentral Texas, used since the early 1960's as a fire fighting training facility. Until 1980, ignitants used on training structures included refinery waste products. Since then, only diesel fuel has been used. Two sludge retention ponds located on the training school grounds collect run-off such as ignitants, flame retardants, fire-fighting chemicals, and water from the practice structures. Chemical analysis of the ponds indicated presence of a number of compounds including partially combusted hydrocarbons, PCB'S, and several heavy metals. Water extracts from the retention ponds gave positive responses in the Salmonella/mammalian microsome assay and the Bacillus DNA Repair Assay, indicating that compounds present in waste water were mutagenic. Peromyscus trapped around the banks of the retention ponds showed significantly increased levels of somatic metaphase chromosome aberrations. The most likely route of exposure for these animals was through ingestion of soil particles during foraging and grooming. Of metals found at the site, at least four (cadmium, chromium, lead, and zinc) have been shown to induce chromosome aberrations in mammalian cell.« less

Idaho National Engineering Laboratory code assessment of the Rocky Flats transuranic waste

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1995-07-01

This report is an assessment of the content codes associated with transuranic waste shipped from the Rocky Flats Plant in Golden, Colorado, to INEL. The primary objective of this document is to characterize and describe the transuranic wastes shipped to INEL from Rocky Flats by item description code (IDC). This information will aid INEL in determining if the waste meets the waste acceptance criteria (WAC) of the Waste Isolation Pilot Plant (WIPP). The waste covered by this content code assessment was shipped from Rocky Flats between 1985 and 1989. These years coincide with the dates for information available in themore » Rocky Flats Solid Waste Information Management System (SWIMS). The majority of waste shipped during this time was certified to the existing WIPP WAC. This waste is referred to as precertified waste. Reassessment of these precertified waste containers is necessary because of changes in the WIPP WAC. To accomplish this assessment, the analytical and process knowledge available on the various IDCs used at Rocky Flats were evaluated. Rocky Flats sources for this information include employee interviews, SWIMS, Transuranic Waste Certification Program, Transuranic Waste Inspection Procedure, Backlog Waste Baseline Books, WIPP Experimental Waste Characterization Program (headspace analysis), and other related documents, procedures, and programs. Summaries are provided of: (a) certification information, (b) waste description, (c) generation source, (d) recovery method, (e) waste packaging and handling information, (f) container preparation information, (g) assay information, (h) inspection information, (i) analytical data, and (j) RCRA characterization.« less

Linamarase activities in Bacillus spp. responsible for thermophilic aerobic digestion of agricultural wastes for animal nutrition.

PubMed

Ugwuanyi, J Obeta; Harvey, L M; McNeil, B

2007-01-01

Thermophilic Bacillus spp. isolated from thermophilic aerobic digestion (TAD) of model agricultural slurry were screened for ability to secret linamarase activity and degrade linamarin, a cyanogenic glycoside toxin abundant in cassava. Screening was performed by both linamarin - picrate assay and by p-nitrophenyl beta-D-glucoside (PNPG) degradation, and results of both assays were related. Linamarase positive isolates were identified as Bacillus coagulans, Bacillus licheniformis and Bacillus stearothermophilus. Enzyme production was growth related and peak production was reached in 48 h in B. coagulans and 36 h in B. stearothermophilus. B. coagulans produced over 40 times greater activity than B. stearothermophilus. Enzyme productivity in shake flask was not strictly related to screening assay result. Crude enzyme of B. coagulans was optimally active at 75 degrees C while that of B. stearothermophilus was optimally active at 80 degrees C and both had optimum activity at pH 8.0. The thermophilic and neutrophilic- to marginally alkaline activity of the crude enzymes could be very useful in the detoxification and reprocessing of cyanogens containing cassava wastes by TAD for use in animal nutrition.

Myostatin promotes the wasting of human myoblast cultures through promoting ubiquitin-proteasome pathway-mediated loss of sarcomeric proteins.

PubMed

Lokireddy, Sudarsanareddy; Mouly, Vincent; Butler-Browne, Gillian; Gluckman, Peter D; Sharma, Mridula; Kambadur, Ravi; McFarlane, Craig

2011-12-01

Myostatin is a negative regulator of skeletal muscle growth and in fact acts as a potent inducer of "cachectic-like" muscle wasting in mice. The mechanism of action of myostatin in promoting muscle wasting has been predominantly studied in murine models. Despite numerous reports linking elevated levels of myostatin to human skeletal muscle wasting conditions, little is currently known about the signaling mechanism(s) through which myostatin promotes human skeletal muscle wasting. Therefore, in this present study we describe in further detail the mechanisms behind myostatin regulation of human skeletal muscle wasting using an in vitro human primary myotube atrophy model. Treatment of human myotube populations with myostatin promoted dramatic myotubular atrophy. Mechanistically, myostatin-induced myotube atrophy resulted in reduced p-AKT concomitant with the accumulation of active dephosphorylated Forkhead Box-O (FOXO1) and FOXO3. We further show that addition of myostatin results in enhanced activation of atrogin-1 and muscle-specific RING finger protein 1 (MURF1) and reduced expression of both myosin light chain (MYL) and myosin heavy chain (MYH). In addition, we found that myostatin-induced loss of MYL and MYH proteins is dependent on the activity of the proteasome and mediated via SMAD3-dependent regulation of FOXO1 and atrogin-1. Therefore, these data suggest that the mechanism through which myostatin promotes muscle wasting is very well conserved between species, and that myostatin-induced human myotube atrophy is mediated through inhibition of insulin-like growth factor (IGF)/phosphoinositide 3-kinase (PI3-K)/AKT signaling and enhanced activation of the ubiquitin-proteasome pathway and elevated protein degradation.

Decolorization of the azo dye Acid Orange 51 by laccase produced in solid culture of a newly isolated Trametes trogii strain.

PubMed

Daâssi, Dalel; Zouari-Mechichi, Hela; Frikha, Fakher; Martinez, Maria Jesus; Nasri, Moncef; Mechichi, Tahar

2013-04-01

This study concerns the decolorization and detoxification of the azo dye Acid Orange 51 (AO51) by crude laccase from Trametes trogii produced in solid culture using sawdust as support media. A three-level Box-Behnken factorial design with four factors (enzyme concentration, 1-hydroxybenzotriazole (HBT) concentration, dye concentration and reaction time) combined with response surface methodology was applied to optimize AO51 decolorization. A mathematical model was developed showing the effect of each factor and their interactions on color removal. The model predicted that Acid Orange 51 decolorization above 87.87 ± 1.27 % could be obtained when enzyme concentration, HBT concentration, dye concentration and reaction time were set at 1 U/mL, 0.75 mM, 60 mg/L and 2 days, respectively. The experimental values were in good agreement with the predicted ones and the models were highly significant, the correlation coefficient (R 2 ) being 0.9. Then the desirability function was employed to determine the optimal decolorization condition for each dye and minimize the process cost simultaneously. In addition, germination index assay showed that laccase-treated dye was detoxified; however in the presence of HBT, the phytotoxicity of the treated dye was increased. By using cheap agro-industrial wastes, such as sawdust, a potential laccase was obtained. The low cost of laccase production may further broaden its application in textile wastewater treatment.

Optimisation of surfactant decontamination and pre-treatment of waste chicken feathers by using response surface methodology.

PubMed

Tesfaye, Tamrat; Sithole, Bruce; Ramjugernath, Deresh; Ndlela, Luyanda

2018-02-01

Commercially processed, untreated chicken feathers are biologically hazardous due to the presence of blood-borne pathogens. Prior to valorisation, it is crucial that they are decontaminated to remove the microbial contamination. The present study focuses on evaluating the best technologies to decontaminate and pre-treat chicken feathers in order to make them suitable for valorisation. Waste chicken feathers were washed with three surfactants (sodium dodecyl sulphate) dimethyl dioctadecyl ammonium chloride, and polyoxyethylene (40) stearate) using statistically designed experiments. Process conditions were optimised using response surface methodology with a Box-Behnken experimental design. The data were compared with decontamination using an autoclave. Under optimised conditions, the microbial counts of the decontaminated and pre-treated chicken feathers were significantly reduced making them safe for handling and use for valorisation applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

Epigenetic down-regulated DDX10 promotes cell proliferation through Akt/NF-κB pathway in ovarian cancer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gai, Muhuizi; Bo, Qifang; Qi, Lixia, E-mail: lixiaqi_dph@sina.com

Ovarian cancer contributes to the majority of ovarian cancer, while the molecular mechanisms remain elusive. Recently, some DEAD box protein 1 has been reported play a tumor suppressor role in ovarian cancer progression. However, the functions of DEAD box protein (DDX) members in ovarian cancer development remain largely unknown. In current study, we retrieved GEO databases and surprisingly found that DDX10 is significantly down-regulated in ovarian cancer tissues compared with normal ovary. These findings suggest that DDX10 might also play a suppressive role in ovarian cancer. We then validated the down-regulated expression pattern of DDX10 in fresh ovarian cancer tissues.more » Furthermore, both loss- and gain-functions assays reveal that the down-regulated DDX10 could promote ovarian cancer proliferation in vitro and the xenograft subcutaneous tumor formation assays confirmed these findings in vivo. In addition, we found that DDX10 is epigenetic silenced by miR-155-5p in ovarian cancer. Moreover, we further preliminary illustrated that down-regulated DDX10 promotes ovarian cancer cell proliferation through Akt/NF-κB pathway. Taken together, in current study, we found a novel tumor suppressor, DDX10, is epigenetic silenced by miR-155-5p in ovarian cancer, and the down-regulated expression pattern of DDX10 promotes ovarian cancer proliferation through Akt/NF-κB pathway. Our findings shed the light that DDX families might be a novel for ovarian cancer treatment. - Highlights: • A novel DEAD box protein, DDX10 is significantly down-regulated in ovarian cancer tissues. • Down-regulated DDX10 promotes ovarian cancer cell proliferation and growth both in vitro and in vivo. • miR-155-5p is highly expressed in ovarian cancer tissues and epigenetically targets DDX10. • DDX10 and miR-155-5p regulates Akt/p65 axis in ovarian cancer cells.« less

Biodegradation of bilge waste from Patagonia with an indigenous microbial community.

PubMed

Nievas, M L; Commendatore, M G; Olivera, N L; Esteves, J L; Bucalá, V

2006-12-01

Oily residues that are generated in normal ship operation are considered hazardous wastes. A biodegradation assay with autochthonous microbiota of Bilge Waste Oily Phase (BWOP) was performed in a bioreactor under controlled conditions. Petroleum, diesel oil, and PAH degraders were isolated from bilge wastes. These bacteria belong to the genus Pseudomonas and are closely related to Pseudomonas stutzeri as shown by 16S rDNA phylogenetic analysis. The indigenous microbial community of the bilge waste was capable of biodegrading the BWOP (1% v/v) with biodegradation efficiencies of 70% for hexane extractable material (HEM), 68% for total hydrocarbons (TH) and 90% for total aromatics hydrocarbons (TA) in 14 days. Solid phase microextraction (SPME) was successfully applied to evaluate hydrocarbon evaporation in a control experiment and demonstrated a mass balance closure of 88%. The SPME and biodegradation results give useful information to improve and scale up the process for BWOP treatment.

Kinetic modelling of anaerobic hydrolysis of solid wastes, including disintegration processes.

PubMed

García-Gen, Santiago; Sousbie, Philippe; Rangaraj, Ganesh; Lema, Juan M; Rodríguez, Jorge; Steyer, Jean-Philippe; Torrijos, Michel

2015-01-01

A methodology to estimate disintegration and hydrolysis kinetic parameters of solid wastes and validate an ADM1-based anaerobic co-digestion model is presented. Kinetic parameters of the model were calibrated from batch reactor experiments treating individually fruit and vegetable wastes (among other residues) following a new protocol for batch tests. In addition, decoupled disintegration kinetics for readily and slowly biodegradable fractions of solid wastes was considered. Calibrated parameters from batch assays of individual substrates were used to validate the model for a semi-continuous co-digestion operation treating simultaneously 5 fruit and vegetable wastes. The semi-continuous experiment was carried out in a lab-scale CSTR reactor for 15 weeks at organic loading rate ranging between 2.0 and 4.7 gVS/Ld. The model (built in Matlab/Simulink) fit to a large extent the experimental results in both batch and semi-continuous mode and served as a powerful tool to simulate the digestion or co-digestion of solid wastes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Gray-Box Approach for Thermal Modelling of Buildings for Applications in District Heating and Cooling Networks

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saurav, Kumar; Chandan, Vikas

District-heating-and-cooling (DHC) systems are a proven energy solution that has been deployed for many years in a growing number of urban areas worldwide. They comprise a variety of technologies that seek to develop synergies between the production and supply of heat, cooling, domestic hot water and electricity. Although the benefits of DHC systems are significant and have been widely acclaimed, yet the full potential of modern DHC systems remains largely untapped. There are several opportunities for development of energy efficient DHC systems, which will enable the effective exploitation of alternative renewable resources, waste heat recovery, etc., in order to increasemore » the overall efficiency and facilitate the transition towards the next generation of DHC systems. This motivated the need for modelling these complex systems. Large-scale modelling of DHC-networks is challenging, as it has several components such as buildings, pipes, valves, heating source, etc., interacting with each other. In this paper, we focus on building modelling. In particular, we present a gray-box methodology for thermal modelling of buildings. Gray-box modelling is a hybrid of data driven and physics based models where, coefficients of the equations from physics based models are learned using data. This approach allows us to capture the dynamics of the buildings more effectively as compared to pure data driven approach. Additionally, this approach results in a simpler models as compared to pure physics based models. We first develop the individual components of the building such as temperature evolution, flow controller, etc. These individual models are then integrated in to the complete gray-box model for the building. The model is validated using data collected from one of the buildings at Lule{\\aa}, a city on the coast of northern Sweden.« less

Targeted Phenotypic Screening in Plasmodium falciparum and Toxoplasma gondii Reveals Novel Modes of Action of Medicines for Malaria Venture Malaria Box Molecules

PubMed Central

2018-01-01

ABSTRACT The Malaria Box collection includes 400 chemically diverse small molecules with documented potency against malaria parasite growth, but the underlying modes of action are largely unknown. Using complementary phenotypic screens against Plasmodium falciparum and Toxoplasma gondii, we report phenotype-specific hits based on inhibition of overall parasite growth, apicoplast segregation, and egress or host invasion, providing hitherto unavailable insights into the possible mechanisms affected. First, the Malaria Box library was screened against tachyzoite stage T. gondii and the half-maximal effective concentrations (EC50s) of molecules showing ≥80% growth inhibition at 10 µM were determined. Comparison of the EC50s for T. gondii and P. falciparum identified a subset of 24 molecules with nanomolar potency against both parasites. Thirty molecules that failed to induce acute growth inhibition in T. gondii tachyzoites in a 2-day assay caused delayed parasite death upon extended exposure, with at least three molecules interfering with apicoplast segregation during daughter cell formation. Using flow cytometry and microscopy-based examinations, we prioritized 26 molecules with the potential to inhibit host cell egress/invasion during asexual developmental stages of P. falciparum. None of the inhibitors affected digestive vacuole integrity, ruling out a mechanism mediated by broadly specific protease inhibitor activity. Interestingly, five of the plasmodial egress inhibitors inhibited ionophore-induced egress of T. gondii tachyzoites. These findings highlight the advantage of comparative and targeted phenotypic screens in related species as a means to identify lead molecules with a conserved mode of action. Further work on target identification and mechanism analysis will facilitate the development of antiparasitic compounds with cross-species efficacy. IMPORTANCE The phylum Apicomplexa includes many human and animal pathogens, such as Plasmodium falciparum (human malaria) and Toxoplasma gondii (human and animal toxoplasmosis). Widespread resistance to current antimalarials and the lack of a commercial vaccine necessitate novel pharmacological interventions with distinct modes of action against malaria. For toxoplasmosis, new drugs to effectively eliminate tissue-dwelling latent cysts of the parasite are needed. The Malaria Box antimalarial collection, managed and distributed by the Medicines for Malaria Venture, includes molecules of novel chemical classes with proven antimalarial efficacy. Using targeted phenotypic assays of P. falciparum and T. gondii, we have identified a subset of the Malaria Box molecules as potent inhibitors of plastid segregation and parasite invasion and egress, thereby providing early insights into their probable mode of action. Five molecules that inhibit the egress of both parasites have been identified for further mechanistic studies. Thus, the approach we have used to identify novel molecules with defined modes of action in multiple parasites can expedite the development of pan-active antiparasitic agents. PMID:29359192

Modeling of anaerobic degradation of solid slaughterhouse waste: inhibition effects of long-chain fatty acids or ammonia.

PubMed

Lokshina, L Y; Vavilin, V A; Salminen, E; Rintala, J

2003-01-01

The anaerobic bioconversion of solid poultry slaughterhouse wastes was kinetically investigated. The modified version of simulation model was applied for description of experimental data in mesophilic laboratory digester and assays. Additionally, stages of formation and consumption of long chain fatty acids (LCFA) were included in the model. Batch data on volatile solids, ammonium, acetate, butyrate, propionate, LCFA concentrations, pH level, cumulative volume, and methane partial pressure were used for model calibration. As a reference, the model was used to describe digestion of solid sorted household waste. Simulation results showed that an inhibition of polymer hydrolysis by volatile fatty acids and acetogenesis by NH3 or LCFA could be responsible for the complex system dynamics during degradation of lipid- and protein-rich wastes.

Identification and characterization of a nuclear localization signal of TRIM28 that overlaps with the HP1 box

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moriyama, Tetsuji; Sangel, Percival; Yamaguchi, Hiroki

2015-07-03

Tripartite motif-containing 28 (TRIM28) is a transcription regulator, which forms a repressor complex containing heterochromatin protein 1 (HP1). Here, we report identification of a nuclear localization signal (NLS) within the 462-494 amino acid region of TRIM28 that overlaps with its HP1 binding site, HP1 box. GST-pulldown experiments revealed the interaction of the arginine-rich TRIM28 NLS with various importin α subtypes (α1, α2 and α4). In vitro transport assay demonstrated that nuclear localization of GFP-TRIM28 NLS is mediated by importin αs, in conjunction with importin β1 and Ran. Further, we demonstrated that HP1 and importin αs compete for binding to TRIM28. Together,more » our findings suggest that importin α has an essential role in the nuclear delivery and preferential HP1 interaction of TRIM28. - Highlights: • TRIM28 contains an NLS within the 462-494 amino acid region. • The nuclear import of TRIM28 is mediated by importin α/importin β1. • TRIM28 NLS overlaps with HP1 Box. • HP1 and importin α compete for binding to TRIM28.« less

Transcriptional Activation of Pyoluteorin Operon Mediated by the LysR-Type Regulator PltR Bound at a 22 bp lys Box in Pseudomonas aeruginosa M18

PubMed Central

Wang, Guohao; Xu, Yuquan

2012-01-01

Pseudomonas aeruginosa M18, a rhizosphere-isolated bacterial strain showing strong antifungal activity, can produce secondary metabolites such as phenazine-1-carboxylic acid and pyoluteorin (Plt). The LysR-type transcriptional regulator PltR activates the Plt biosynthesis operon pltLABCDEFG, the expression of which is induced by Plt. Here, we identified and characterized the non-conserved pltL promoter (pltLp) specifically activated by PltR and its upstream neighboring lys box from the complicated pltR–pltL intergenic sequence. The 22 bp palindromic lys box, which consists of two 9 bp complementary inverted repeats interrupted by 4 bp, was found to contain the conserved, GC-rich LysR-binding motif (T-N11-A). Evidence obtained in vivo from mutational and lacZ report analyses and in vitro from electrophoretic mobility shift assays reveals that the PltR protein directly bound to the pltLp region as the indispensable binding motif “lys box”, thereby transcriptionally activating the pltLp-driven plt operon expression. Plt, as a potential non-essential coinducer of PltR, specifically induced the pltLp expression and thus strengthened its biosynthetic plt operon expression. PMID:22761817

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

PubMed

Cho, Seok Keun; Bae, Hansol; Ryu, Moon Young; Wook Yang, Seong; Kim, Woo TaeK

2015-09-04

Drought stress strongly affects plant growth and development, directly connected with crop yields, accordingly. However, related to the function of U-BOX E3 ligases, the underlying molecular mechanisms of desiccation stress response in plants are still largely unknown. Here we report that PUB22 and PUB23, two U-box E3 ligase homologs, tether ubiquitins to 19S proteasome regulatory particle (RP) subunit RPN6, leading to its degradation. RPN6 was identified as an interacting substrate of PUB22 by yeast two-hybrid screening, and in vitro pull-down assay confirmed that RPN6 interacts not only with PUB22, but also with PUB23. Both PUB22 and PUB23 were able to conjugate ubiquitins on RPN6 in vitro. Furthermore, RPN6 showed a shorter protein half-life in PUB22 overexpressing plants than in wild-type, besides RPN6 was significantly stabilized in pub22pub23 double knockout plants. Taken together, these results solidify a notion that PUB22 and PUB23 can alter the activity of 26S proteasome in response to drought stress. Copyright © 2015 Elsevier Inc. All rights reserved.

TonB-dependent ligand trapping in the BtuB transporter.

PubMed

Mills, Allan; Le, Hai-Tuong; Duong, Franck

2016-12-01

TonB-dependent transporters are β-barrel outer membrane proteins occluded by a plug domain. Upon ligand binding, these transporters extend a periplasmic motif termed the TonB box. The TonB box permits the recruitment of the inner membrane protein complex TonB-ExbB-ExbD, which drives import of ligands in the cell periplasm. It is unknown precisely how the plug domain is moved aside during transport nor have the intermediate states between TonB recruitment and plug domain movement been characterized biochemically. Here we employ nanodiscs, native gel electrophoresis, and scintillation proximity assays to determine the binding kinetics of vitamin B 12 to BtuB. The results show that ligand-bound BtuB recruits a monomer of TonB (TonB ∆1-31 ), which in turn increases retention of vitamin B 12 within the transporter. The TonB box and the extracellular residue valine 90 that forms part of the vitamin B 12 binding site are essential for this event. These results identify a novel step in the TonB-dependent transport process. They show that TonB binding to BtuB trap the ligand, possibly until the ExbB-ExbD complex is activated or recruited to ensure subsequent transport. Copyright © 2016 Elsevier B.V. All rights reserved.

The problem of anti-doping control of luteinizing hormone in boxing.

PubMed

Llouquet, Jean Louis; Crepin, Nathalie; Lasne, Françoise

2013-04-01

Luteinizing hormone (LH) is physiologically produced by the anterior pituitary gland. Male athletes may use pharmaceutical LH for doping since it increases the production of testosterone by testes. This hormone is thus on the World Anti-Doping Agency (WADA) list of substances prohibited for males. Anti-doping laboratories perform the assay of this hormone in urine and report abnormally elevated results. We observed a highly significant prevalence of abnormal results in samples taken after a boxing match. Comparison of the descriptive statistics for 426 LH values observed in boxing and other sports showed significant differences. An experimental study comparing urinary LH levels in 17 boxers before and after a match demonstrated a clear increase after the match. The same observation was made for urinary follicle stimulating hormone (FSH) in all of the eight boxers tested for this other pituitary gonadotropin. These observations have consequences for anti-doping controls, as the reference range for urinary LH levels must take into account the specificities of boxers. They also suggest consequences for the health of boxers. Although to our knowledge such observations have never been described, other pituitary disorders have been reported. Our results deserve further investigation from a medical point of view. Copyright © 2013 John Wiley & Sons, Ltd.

Effect of channelling on water balance, oxygen diffusion and oxidation rate in mine waste rock with an inclined multilayer soil cover

NASA Astrophysics Data System (ADS)

Song, Qing; Yanful, Ernest K.

2010-05-01

Engineered soil covers provide an option to mitigate acid rock drainage through reduced water flow and gaseous oxygen influx to underlying mine waste. Channels such as fissures, cracks or fractures developed in the barrier may influence the long-term performance of the soil cover. However, limited published information is available on the extent to which soil cover performance is impacted by these fissures and cracks. This study was conducted to investigate the effect of channelling in a barrier layer on water flow and oxygen transport in a soil cover. Two inclined (a slope of 20%) multilayer soil covers were examined under laboratory conditions. One cover had a 10-cm wide sand-filled channel in a compacted barrier layer (silty clay) at the upslope section, while the other cover was a normal one without the channel pathway. The soil covers were installed in plastic boxes measuring 120 cm × 120 cm × 25 cm (width × height × thickness). The sand-filled channel was designed to represent the aggregate of fissures and cracks that may be present in the compacted barrier. The soil covers were subjected to controlled drying and wetting periods selected to simulate field situation at the Whistle mine site near Capreol, Ontario, Canada. The measured results indicated that interflow decreased from 72.8% of the total precipitation in the soil cover without channel flow to 35.3% in the cover with channel flow, and percolation increased from zero in the normal soil cover to 43.0% of the total precipitation in the cover with channel flow. Gaseous oxygen transfer into the waste rock below the cover soils was 1091 times greater in the cover with channel than in the soil cover without channel. The channel pathway present in the barrier layer acted as a major passage for water movement and gaseous oxygen diffusion into the waste rock layer, thus decreasing the performance of the soil cover. The spacing of the channel with respect to the length of the test box is similar to those found in other published fracture networks. The distribution and partitioning of the water balance components would be expected to be similar to other situations with the same cover slope. This, of course, would depend on rainfall intensity.

Nicotiana tabacum EIL2 directly regulates expression of at least one tobacco gene induced by sulphur starvation.

PubMed

Wawrzyńska, Anna; Lewandowska, Małgorzata; Sirko, Agnieszka

2010-03-01

Sulphur deficiency severely affects plant growth and their agricultural productivity leading to diverse changes in development and metabolisms. Molecular mechanisms regulating gene expression under low sulphur conditions remain largely unknown. AtSLIM1, a member of the EIN3-like (EIL) family was reported to be a central transcriptional regulator of the plant sulphur response, however, no direct interaction of this protein with any sulphur-responsive promoters was demonstrated. The focus of this study was on the analysis of a promoter region of UP9C, a tobacco gene strongly induced by sulphur limitation. Cloning and subsequent examination of this promoter resulted in the identification of a 20-nt sequence (UPE-box), also present in the promoters of several Arabidopsis genes, including three out of four homologues of UP9C. The UPE-box, consisting of two parallel tebs sequences (TEIL binding site), proved to be necessary to bind the transcription factors belonging to the EIL family and of a 5-nt conserved sequence at the 3'-end. The yeast one-hybrid analysis resulted in the identification of one transcription factor (NtEIL2) capable of binding to the UPE-box. The interactions of NtEIL2, and its homologue from Arabidopsis, AtSLIM1, with DNA were affected by mutations within the UPE-box. Transient expression assays in Nicotiana benthamiana have further shown that both factors, NtEIL2 and AtSLIM1, activate the UP9C promoter. Interestingly, activation by NtEIL2, but not by AtSLIM1, was dependent on the sulphur-deficiency of the plants.

Downregulation of MicroRNA 29a/b exacerbated diabetic retinopathy by impairing the function of Müller cells via Forkhead box protein O4.

PubMed

Zhang, Jiayu; Wu, Liang; Chen, Jiawei; Lin, Sisi; Cai, Daqiu; Chen, Chengwei; Chen, Zhenguo

2018-05-01

Diabetic retinopathy is a neurological disease, which can lead to blindness in severe cases. The pathogenesis underlying diabetic retinopathy is unclear. The aim of this study was to explore the role of dysregulated microRNA 29a/b in the onset and progression of diabetic retinopathy. Diabetes mellitus was induced in rats using 60 mg/kg of streptozotocin. Glucose (5.5 and 25 mM) was used to stimulate rat retinal Müller cells. Real-time polymerase chain reaction and Western blot analyses were used to determine gene expression. A luciferase reporter assay was conducted to validate the relationship of microRNA 29a/b with glioma-associated oncogene homolog 1 and Forkhead box protein O4. The expression of microRNA 29a/b and glutamine synthetase decreased in both diabetes mellitus rats and rat retinal Müller cells stimulated with high glucose, whereas the expression of sonic hedgehog, glioma-associated oncogene homolog 1, glial fibrillary acidic protein, and vascular endothelial growth factor, as well as the content of glutamate, increased. Dysregulated microRNA 29a/b was directly regulated by the sonic hedgehog-glioma-associated oncogene homolog 1 signalling pathway, and microRNA 29a and microRNA 29b targeted Forkhead box protein O4 and regulated its expression. Downregulation of microRNA 29a/b, mediated by the sonic hedgehog-glioma-associated oncogene homolog 1 signalling pathway, exacerbated diabetic retinopathy by upregulating Forkhead box protein O4.

Signal Transduction Pathway in Maspin-induced Tumor Suppression of Prostate Cancer

DTIC Science & Technology

2002-03-01

the zip Ebr allele is tested in similar assays with BR-C or Sb-sbd mutants. The zipEbr mutation is associated with a missense alteration in the myosin ...cytoskeletal dynamics in elongating legs via by inducing contraction of the apical actin- myosin belt. 8 Recent evidence has shown that mutations in...the RhoA mutations used in these studies have been characterized at a molecular level (Table 2). RhoAE3 °o is a CAAX box missense mutation (C to Y

Tortoiseshell or Polymer? Spectroscopic Analysis to Redefine a Purported Tortoiseshell Box with Gold Decorations as a Plastic Box with Brass.

PubMed

Pereira, António; Caldeira, Ana Teresa; Maduro, Belmira; Vandenabeele, Peter; Candeias, António

2016-01-01

The study and preservation of museum collections requires complete knowledge and understanding of constituent materials that can be natural, synthetic, or semi-synthetic polymers. In former times, objects were incorporated in museum collections and classified solely by their appearance. New studies, prompted by severe degradation processes or conservation-restoration actions, help shed light on the materiality of objects that can contradict the original information or assumptions. The selected case study presented here is of a box dating from the beginning of the 20th century that belongs to the Portuguese National Ancient Art Museum. Museum curators classified it as a tortoiseshell box decorated with gold applications solely on the basis of visual inspection and the information provided by the donor. This box has visible signs of degradation with white veils, initially assumed to be the result of biological degradation of a proteinaceous matrix. This paper presents the methodological rationale behind this study and proposes a totally non-invasive methodology for the identification of polymeric materials in museum artifacts. The analysis of surface leachates using (1)H and (13)C nuclear magnetic resonance (NMR) complemented by in situ attenuated total reflection infrared spectroscopy (ATR FT-IR) allowed for full characterization of the object s substratum. The NMR technique unequivocally identified a great number of additives and ATR FT-IR provided information about the polymer structure and while also confirming the presence of additives. The pressure applied during ATR FT-IR spectroscopy did not cause any physical change in the structure of the material at the level of the surface (e.g., color, texture, brightness, etc.). In this study, variable pressure scanning electron microscopy (VP-SEM-EDS) was also used to obtain the elemental composition of the metallic decorations. Additionally, microbiologic and enzymatic assays were performed in order to identify the possible biofilm composition and understand the role of microorganisms in the biodeterioration process. Using these methodologies, the box was correctly identified as being made of cellulose acetate plastic with brass decorations and the white film was identified as being composed mainly of polymer exudates, namely sulphonamides and triphenyl phosphate. © The Author(s) 2015.

Integration of GC-MSD and ER-Calux® assay into a single protocol for determining steroid estrogens in environmental samples.

PubMed

Avberšek, Miha; Žegura, Bojana; Filipič, Metka; Heath, Ester

2011-11-01

There are many published studies that use either chemical or biological methods to investigate steroid estrogens in the aquatic environment, but rarer are those that combine both. In this study, gas chromatography with mass selective detection (GC-MSD) and the ER-Calux(®) estrogenicity assay were integrated into a single protocol for simultaneous determination of natural (estrone--E1, 17β-estradiol--E2, estriol--E3) and synthetic (17α-ethinylestradiol--EE2) steroid estrogens concentrations and the total estrogenic potential of environmental samples. For integration purposes, several solvents were investigated and the commonly used dimethyl sulphoxide (DMSO) in the ER-Calux(®) assay was replaced by ethyl acetate, which is more compatible with gas chromatography and enables the same sample to be analysed by both GC-MSD and the ER-Calux(®) assay. The integrated protocol was initially tested using a standard mixture of estrogens. The results for pure standards showed that the estrogenicity calculated on the basis of GC-MSD and the ER-Calux(®) assay exhibited good correlation (r(2)=0.96; α=0.94). The result remained the same when spiked waste water extracts were tested (r(2)=0.92, α=1.02). When applied to real waste water influent and effluent samples the results proved (r(2)=0.93; α=0.99) the applicability of the protocol. The main advantages of this newly developed protocol are simple sample handling for both methods, and reduced material consumption and labour. In addition, it can be applied as either a complete or sequential analysis where the ER-Calux(®) assay is used as a pre-screening method prior to the chemical analysis. Copyright © 2011 Elsevier B.V. All rights reserved.

Radiation Measurement Systems and Experiences in Japan after the Fukushima Accident

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

In the approximately 4 years since the March 2011 earthquake and subsequent Tsunami damaged the three Dai-Ichi nuclear power plants, the AREVA and Canberra team have deployed a variety of different radiation measurement machines in Japan. These measurement projects or instruments include: - Dose-rate mapping of the close-in areas on the damaged reactor site using the stored output of dose rate meters carried by workers planning for the installing of a water processing unit; - Spectroscopic on-line measurements of the first water processing unit on the site, where these nuclide-specific measurements at multiple points allowed remote users to determine themore » optimum processing parameters - General purpose food measurement units, both high precision HPGe systems and low cost NaI systems, which were used throughout the country for food quality measurements; - In-vivo measurement systems, both fixed and mobile whole body counting units, that were used on at least 500,000 people, demonstrating to them that internal doses were very low; - In-vivo measurements with special high-sensitivity whole body counters on babies and children, demonstrating to the parents of these children that internal doses are very low; - Automatic systems for high throughput assay of bags of rice that are part of the consumer confidence plan to prove to customers that Fukushima Prefecture rice is safe; - High sensitivity automatic system to assay boxes of special semi-dry persimmons and to report the activity of each individual item within the box meets the regulatory limit; - System to assay soil on a conveyor belt and sort the output according to level of radioactivity, which would then reduce the volume of material that must be treated as radioactive; - System to assay a truck loaded with 1 cubic meter sacks of soil and vegetation, and report the results of each individual sack. - On-line water measurement system for SrY90 at levels that are suitable for release to the environment; - Mobile system to prove that rolls of grass harvested for animal feed meet the regulatory requirements. This document briefly describes the purpose of each measurement project, describes the instruments and assay method that used, and discusses some of the operational experiences from each of these projects. (authors)« less

A HIV-1 Tat mutant protein disrupts HIV-1 Rev function by targeting the DEAD-box RNA helicase DDX1.

PubMed

Lin, Min-Hsuan; Sivakumaran, Haran; Jones, Alun; Li, Dongsheng; Harper, Callista; Wei, Ting; Jin, Hongping; Rustanti, Lina; Meunier, Frederic A; Spann, Kirsten; Harrich, David

2014-12-14

Previously we described a transdominant negative mutant of the HIV-1 Tat protein, termed Nullbasic, that downregulated the steady state levels of unspliced and singly spliced viral mRNA, an activity caused by inhibition of HIV-1 Rev activity. Nullbasic also altered the subcellular localizations of Rev and other cellular proteins, including CRM1, B23 and C23 in a Rev-dependent manner, suggesting that Nullbasic may disrupt Rev function and trafficking by intervening with an unidentified component of the Rev nucleocytoplasmic transport complex. To seek a possible mechanism that could explain how Nullbasic inhibits Rev activity, we used a proteomics approach to identify host cellular proteins that interact with Nullbasic. Forty-six Nullbasic-binding proteins were identified by mass spectrometry including the DEAD-box RNA helicase, DDX1. To determine the effect of DDX1 on Nullbasic-mediated Rev activity, we performed cell-based immunoprecipitation assays, Rev reporter assays and bio-layer interferometry (BLI) assays. Interaction between DDX1 and Nullbasic was observed by co-immunoprecipitation of Nullbasic with endogenous DDX1 from cell lysates. BLI assays showed a direct interaction between Nullbasic and DDX1. Nullbasic affected DDX1 subcellular distribution in a Rev-independent manner. Interestingly overexpression of DDX1 in cells not only restored Rev-dependent mRNA export and gene expression in a Rev reporter assay but also partly reversed Nullbasic-induced Rev subcellular mislocalization. Moreover, HIV-1 wild type Tat co-immunoprecipitated with DDX1 and overexpression of Tat could rescue the unspliced viral mRNA levels inhibited by Nullbasic in HIV-1 expressing cells. Nullbasic was used to further define the complex mechanisms involved in the Rev-dependent nuclear export of the 9 kb and 4 kb viral RNAs. All together, these data indicate that DDX1 can be sequestered by Nullbasic leading to destabilization of the Rev nucleocytoplasmic transport complex and decreased levels of Rev-dependent viral transcripts. The outcomes support a role for DDX1 in maintenance of a Rev nuclear complex that transports viral RRE-containing mRNA to the cytoplasm. To our knowledge Nullbasic is the first anti-HIV protein that specifically targets the cellular protein DDX1 to block Rev's activity. Furthermore, our research raises the possibility that wild type Tat may play a previously unrecognized but very important role in Rev function.

Evaluation of Bacterial Expansin EXLX1 as a Cellulase Synergist for the Saccharification of Lignocellulosic Agro-Industrial Wastes

PubMed Central

Lin, Hui; Shen, Qi; Zhan, Ju-Mei; Wang, Qun; Zhao, Yu-Hua

2013-01-01

Various types of lignocellulosic wastes extensively used in biofuel production were provided to assess the potential of EXLX1 as a cellulase synergist. Enzymatic hydrolysis of natural wheat straw showed that all the treatments using mixtures of cellulase and an optimized amount of EXLX1, released greater quantities of sugars than those using cellulase alone, regardless of cellulase dosage and incubation time. EXLX1 exhibited different synergism and binding characteristics for different wastes, but this can be related to their lignocellulosic components. The cellulose proportion could be one of the important factors. However, when the cellulose proportion of different biomass samples exhibited no remarkable differences, a higher synergism of EXLX1 is prone to occur on these materials, with a high proportion of hemicellulose and a low proportion of lignin. The information could be favorable to assess whether EXLX1 is effective as a cellulase synergist for the hydrolysis of the used materials. Binding assay experiments further suggested that EXLX1 bound preferentially to alkali pretreated materials, as opposed to acid pretreated materials under the assay condition and the binding preference would be affected by incubation temperature. PMID:24086425

Tank Closure Progress at the Department of Energy's Idaho National Engineering Laboratory Tank Farm Facility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Quigley, K.D.; Butterworth, St.W.; Lockie, K.A.

2008-07-01

Significant progress has been made at the U.S. Department of Energy (DOE) Idaho National Laboratory (INL) to empty, clean and close radioactive liquid waste storage tanks at the Idaho Nuclear Technology and Engineering Center (INTEC) Tank Farm Facility (TFF). The TFF includes eleven 1,135.6-kL (300,000-gal) underground stainless steel storage tanks and four smaller, 113.5-kL (30,000-gal) stainless steel tanks, along with tank vaults, interconnecting piping, and ancillary equipment. The TFF tanks have historically been used to store a variety of radioactive liquid waste, including wastes associated with past spent nuclear fuel reprocessing. Although four of the large storage tanks remain inmore » use for waste storage, the other seven 1,135.6-kL (300,000-gal) tanks and the four 113.5-kL (30,000-gal) tanks have been emptied of waste, cleaned and filled with grout. A water spray cleaning system was developed and deployed to clean internal tank surfaces and remove remaining tank wastes. The cleaning system was effective in removing all but a very small volume of solid residual waste particles. Recent issuance of an Amended Record of Decision (ROD) in accordance with the National Environmental Policy Act, and a Waste Determination complying with Section 3116 of the Ronald W. Reagan National Defense Authorization Act (NDAA) for Fiscal Year 2005, has allowed commencement of grouting activities on the cleaned tanks. The first three 113.5-kL (30,000-gal) tanks were grouted in the Fall of 2006 and the fourth tank and the seven 1,135.6-kL (300,000-gal) tanks were filled with grout in 2007 to provide long-term stability. It is currently planned that associated tank valve boxes and interconnecting piping, will be stabilized with grout as early as 2008. (authors)« less

Minimum airflow reset of single-duct VAV terminal boxes

NASA Astrophysics Data System (ADS)

Cho, Young-Hum

Single duct Variable Air Volume (VAV) systems are currently the most widely used type of HVAC system in the United States. When installing such a system, it is critical to determine the minimum airflow set point of the terminal box, as an optimally selected set point will improve the level of thermal comfort and indoor air quality (IAQ) while at the same time lower overall energy costs. In principle, this minimum rate should be calculated according to the minimum ventilation requirement based on ASHRAE standard 62.1 and maximum heating load of the zone. Several factors must be carefully considered when calculating this minimum rate. Terminal boxes with conventional control sequences may result in occupant discomfort and energy waste. If the minimum rate of airflow is set too high, the AHUs will consume excess fan power, and the terminal boxes may cause significant simultaneous room heating and cooling. At the same time, a rate that is too low will result in poor air circulation and indoor air quality in the air-conditioned space. Currently, many scholars are investigating how to change the algorithm of the advanced VAV terminal box controller without retrofitting. Some of these controllers have been found to effectively improve thermal comfort, indoor air quality, and energy efficiency. However, minimum airflow set points have not yet been identified, nor has controller performance been verified in confirmed studies. In this study, control algorithms were developed that automatically identify and reset terminal box minimum airflow set points, thereby improving indoor air quality and thermal comfort levels, and reducing the overall rate of energy consumption. A theoretical analysis of the optimal minimum airflow and discharge air temperature was performed to identify the potential energy benefits of resetting the terminal box minimum airflow set points. Applicable control algorithms for calculating the ideal values for the minimum airflow reset were developed and applied to actual systems for performance validation. The results of the theoretical analysis, numeric simulations, and experiments show that the optimal control algorithms can automatically identify the minimum rate of heating airflow under actual working conditions. Improved control helps to stabilize room air temperatures. The vertical difference in the room air temperature was lower than the comfort value. Measurements of room CO2 levels indicate that when the minimum airflow set point was reduced it did not adversely affect the indoor air quality. According to the measured energy results, optimal control algorithms give a lower rate of reheating energy consumption than conventional controls.

Particle Swarm Imaging (PSIM) - Innovative Gamma-Ray Assay - 13497

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parvin, Daniel; Clarke, Sean; Humes, Sarah J.

2013-07-01

Particle Swarm Imaging is an innovative technique used to perform quantitative gamma-ray assay. The innovation overcomes some of the difficulties associated with the accurate measurement and declaration of measurement uncertainties of radionuclide inventories within waste items when the distribution of activity is unknown. Implementation requires minimal equipment, with field measurements and results obtained using only a single electrically cooled HRGS gamma-ray detector. Examples of its application in the field are given in this paper. (authors)

Effects of Holding Time, Storage, and the Preservation of ...

EPA Pesticide Factsheets

The purpose of this project was to answer questions related to storage of samples to be analyzed by the quantitative polymerase chain reaction (qPCR)-based assays for fecal indicator bacteria. The project was divided into two parts. The first part was to determine if filters that were used to collect fecal indicators could be stored frozen and analyzed at a later date and the second part was to determine if refrigerated water samples could be held for 24 to 48 hours prior to analysis by qPCR. Both of these studies answer questions that were important in the analysis of fresh and marine surface water samples for beach monitoring purposes. 1) Develop and evaluate qPCR assays and test methods for the detection and quantification of genetic markers from indicator bacteria that are associated with human fecal waste and from two new groups of general fecal indicator bacteria (E. coli and Clostridia) that historically have been widely used or are favored in specific regions 2) Determine the occurrence and densities of genetic markers detected by new qPCR assays developed under objective 1 and compare with occurrence and densities of genetic markers detected by previously developed qPCR assays for enterococci and total Bacterioidalesin waste waters and fecal material from different animal sources. 3) Determine stability of fecal indicator bacteria target DNA sequences in freezer archived filter retentates of ambient surface water samples 4) Determine the densitie

Duplex real-time PCR assay using SYBR Green to detect and quantify Malayan box turtle (Cuora amboinensis) materials in meatballs, burgers, frankfurters and traditional Chinese herbal jelly powder.

PubMed

Asing; Ali, Eaqub; Hamid, Sharifah Bee Abd; Hossain, Motalib; Ahamad, Mohammad Nasir Uddin; Hossain, S M Azad; Naquiah, Nina; Zaidul, I S M

2016-11-01

The Malayan box turtle (Cuora amboinensis) (MBT) is a vulnerable and protected species widely used in exotic foods and traditional medicines. Currently available polymerase chain reaction (PCR) assays to identify MBT lack automation and involve long targets which break down in processed or denatured tissue. This SYBR Green duplex real-time PCR assay has addressed this research gap for the first time through the combination of 120- and 141-bp targets from MBT and eukaryotes for the quantitative detection of MBT DNA in food chain and herbal medicinal preparations. This authentication ensures better security through automation, internal control and short targets that were stable under the processing treatments of foods and medicines. A melting curve clearly demonstrated two peaks at 74.63 ± 0.22 and 78.40 ± 0.31°C for the MBT and eukaryotic products, respectively, under pure, admixed and commercial food matrices. Analysis of 125 reference samples reflected a target recovery of 93.25-153.00%, PCR efficiency of 99-100% and limit of detection of 0.001% under various matrices. The quantification limits were 0.00001, 0.00170 ± 0.00012, 0.00228 ± 0.00029, 0.00198 ± 0.00036 and 0.00191 ± 0.00043 ng DNA for the pure meat, binary mixtures, meatball, burger and frankfurter products, respectively. The assay was used to screen 100 commercial samples of traditional Chinese herbal jelly powder from eight different brands; 22% of them were found to be MBT-positive (5.37 ± 0.50-7.00 ± 0.34% w/w), which was reflected through the Ct values (26.37 ± 0.32-28.90 ± 0.42) and melting curves (74.63-78.65 ± 0.22°C) of the amplified MBT target (120 bp), confirming the speculation that MBT materials are widely used in Chinese herbal desserts, exotic dishes consumed with the hope of prolonging life and youth.

Identification of a Skp1-like protein interacting with SFB, the pollen S determinant of the gametophytic self-incompatibility in Prunus.

PubMed

Matsumoto, Daiki; Yamane, Hisayo; Abe, Kazuyuki; Tao, Ryutaro

2012-07-01

Many species in Rosaceae, Solanaceae, and Plantaginaceae exhibit S-RNase-based self-incompatibility (SI). In this system, the pistil and pollen specificities are determined by S-RNase and the S locus F-box protein, respectively. The pollen S determinant F-box protein in Prunus (Rosaceae) is referred to by two different terms, SFB (for S-haplotype-specific F-box protein) and SLF (for S locus F box), whereas it is called SLF in Solanaceae and Plantaginaceae. Prunus SFB is thought to be a molecule indispensable for its cognate S-RNase to exert cytotoxicity and to arrest pollen tube growth in incompatible reactions. Although recent studies have demonstrated the molecular function of SCF(SLF) in the SI reaction of Solanaceae and Plantaginaceae, how SFB participates in the Prunus SI mechanism remains to be elucidated. Here we report the identification of sweet cherry (Prunus avium) SFB (PavSFB)-interacting Skp1-like1 (PavSSK1) using a yeast (Saccharomyces cerevisiae) two-hybrid screening against the pollen cDNA library. Phylogenetic analysis showed that PavSSK1 belongs to the same clade as Antirrhinum hispanicum SLF-interacting Skp1-like1 and Petunia hybrida SLF-interacting Skp1-like1 (PhSSK1). In yeast, PavSSK1 interacted not only with PavSFBs from different S haplotypes and Cullin1-likes (PavCul1s), but also with S-locus F-box-likes. A pull-down assay confirmed the interactions between PavSSK1 and PavSFB and between PavSSK1 and PavCul1s. These results collectively indicate that PavSSK1 could be a functional component of the SCF complex and that PavSFB may function as a component of the SCF complex. We discuss the molecular function of PavSFB in self-/nonself-recognition in the gametophytic SI of Prunus.

Identification of a Skp1-Like Protein Interacting with SFB, the Pollen S Determinant of the Gametophytic Self-Incompatibility in Prunus1[W

PubMed Central

Matsumoto, Daiki; Yamane, Hisayo; Abe, Kazuyuki; Tao, Ryutaro

2012-01-01

Many species in Rosaceae, Solanaceae, and Plantaginaceae exhibit S-RNase-based self-incompatibility (SI). In this system, the pistil and pollen specificities are determined by S-RNase and the S locus F-box protein, respectively. The pollen S determinant F-box protein in Prunus (Rosaceae) is referred to by two different terms, SFB (for S-haplotype-specific F-box protein) and SLF (for S locus F box), whereas it is called SLF in Solanaceae and Plantaginaceae. Prunus SFB is thought to be a molecule indispensable for its cognate S-RNase to exert cytotoxicity and to arrest pollen tube growth in incompatible reactions. Although recent studies have demonstrated the molecular function of SCFSLF in the SI reaction of Solanaceae and Plantaginaceae, how SFB participates in the Prunus SI mechanism remains to be elucidated. Here we report the identification of sweet cherry (Prunus avium) SFB (PavSFB)-interacting Skp1-like1 (PavSSK1) using a yeast (Saccharomyces cerevisiae) two-hybrid screening against the pollen cDNA library. Phylogenetic analysis showed that PavSSK1 belongs to the same clade as Antirrhinum hispanicum SLF-interacting Skp1-like1 and Petunia hybrida SLF-interacting Skp1-like1 (PhSSK1). In yeast, PavSSK1 interacted not only with PavSFBs from different S haplotypes and Cullin1-likes (PavCul1s), but also with S-locus F-box-likes. A pull-down assay confirmed the interactions between PavSSK1 and PavSFB and between PavSSK1 and PavCul1s. These results collectively indicate that PavSSK1 could be a functional component of the SCF complex and that PavSFB may function as a component of the SCF complex. We discuss the molecular function of PavSFB in self-/nonself-recognition in the gametophytic SI of Prunus. PMID:22548785

Bioremediation of cooking oil waste using lipases from wastes

PubMed Central

do Prado, Débora Zanoni; Facanali, Roselaine; Marques, Márcia Mayo Ortiz; Nascimento, Augusto Santana; Fernandes, Célio Junior da Costa; Zambuzzi, William Fernando

2017-01-01

Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases. PMID:29073166

Bioremediation of cooking oil waste using lipases from wastes.

PubMed

Okino-Delgado, Clarissa Hamaio; Prado, Débora Zanoni do; Facanali, Roselaine; Marques, Márcia Mayo Ortiz; Nascimento, Augusto Santana; Fernandes, Célio Junior da Costa; Zambuzzi, William Fernando; Fleuri, Luciana Francisco

2017-01-01

Cooking oil waste leads to well-known environmental impacts and its bioremediation by lipase-based enzymatic activity can minimize the high cytotoxic potential. In addition, they are among the biocatalysts most commercialized worldwide due to the versatility of reactions and substrates. However, although lipases are able to process cooking oil wastes, the products generated from this process do not necessarily become less toxic. Thus, the aim of the current study is to analyze the bioremediation of lipase-catalyzed cooking oil wastes, as well as their effect on the cytotoxicity of both the oil and its waste before and after enzymatic treatment. Thus, assessed the post-frying modification in soybean oil and in its waste, which was caused by hydrolysis reaction catalyzed by commercial and home-made lipases. The presence of lipases in the extracts obtained from orange wastes was identified by zymography. The profile of the fatty acid esters formed after these reactions was detected and quantified through gas chromatography and fatty acids profile compared through multivariate statistical analyses. Finally, the soybean oil and its waste, with and without enzymatic treatment, were assessed for toxicity in cytotoxicity assays conducted in vitro using fibroblast cell culture. The soybean oil wastes treated with core and frit lipases through transesterification reaction were less toxic than the untreated oils, thus confirming that cooking oil wastes can be bioremediated using orange lipases.

Inferring microbial interactions in thermophilic and mesophilic anaerobic digestion of hog waste

PubMed Central

Shaw, Grace Tzun-Wen; Liu, An-Chi; Weng, Chieh-Yin; Chou, Chu-Yang

2017-01-01

Anaerobic digestion (AnD) is a microbiological process that converts organic waste materials into biogas. Because of its high methane content, biogas is a combustible energy source and serves as an important environmental technology commonly used in the management of animal waste generated on large animal farms. Much work has been done on hardware design and process engineering for the generation of biogas. However, little is known about the complexity of the microbiology in this process. In particular, how microbes interact in the digester and eventually breakdown and convert organic matter into biogas is still regarded as a “black box.” We used 16S rRNA sequencing as a tool to study the microbial community in laboratory hog waste digesters under tightly controlled conditions, and systematically unraveled the distinct interaction networks of two microbial communities from mesophilic (MAnD) and thermophilic anaerobic digestion (TAnD). Under thermophilic conditions, the well-known association between hydrogen-producing bacteria, e.g., Ruminococcaceae and Prevotellaceae, and hydrotrophic methanogens, Methanomicrobiaceae, was reverse engineered by their interactive topological niches. The inferred interaction network provides a sketch enabling the determination of microbial interactive relationships that conventional strategy of finding differential taxa was hard to achieve. This research is still in its infancy, but it can help to depict the dynamics of microbial ecosystems and to lay the groundwork for understanding how microorganisms cohabit in the anaerobic digester. PMID:28732056

Optimization of polyhydroxybutyrate production utilizing waste water as nutrient source by Botryococcus braunii Kütz using response surface methodology.

PubMed

Kavitha, Ganapathy; Kurinjimalar, Chidambaram; Sivakumar, Krishnan; Kaarthik, Muthukumar; Aravind, Rajamani; Palani, Perumal; Rengasamy, Ramasamy

2016-12-01

Investigations have been made to optimize various factors including pH, temperature, and substrate for enhanced polyhydroxybutyrate (PHB) production in Botryococcus braunii which serves as a pioneer for production of bioplastic (PHB). Polyhydroxybutyrate is a natural, decomposable polymers accumulated by the microorganism under different nutritional condition. Strain selection was done by staining method using Sudan black and Nile red dye. Using response surface methodology (RSM), three level- three variables Box Behnken design (BBD), the best potential combination of pH (4-11), temperature (30-50°C) and sewage waste water as substrate fed at different concentrations at 20%-100% for maximum PHB production was investigated. Maximum yield (247±0.42mg/L) of PHB dry weight was achieved from the 60% concentration of sewage waste water as a growth medium at pH 7.5 at 40°C. It was well in close agreement with the value predicted by RSM model yield (246± 0.32mg/L). Thus the study shows the production of PHB by B. braunii along with the basic characterization of PHB by using FTIR and TEM analysis. These preliminary studies indicated that PHB can also be produced by B. braunii utilizing waste water. There is no report on the optimization of PHB production in this microalgae have been documented. Copyright © 2016 Elsevier B.V. All rights reserved.

Bisphenol A and its structural analogues in household waste paper.

PubMed

Pivnenko, K; Pedersen, G A; Eriksson, E; Astrup, T F

2015-10-01

Bisphenol A (BPA) is an industrial chemical produced in large volumes. Its main use is associated with polycarbonate plastic, epoxy resins and thermal paper. In contrast to other applications, thermal paper contains BPA in its un-reacted form as an additive, which is subjected to migration. Receiving a significant amount of attention from the scientific community and beyond, due to its controversial endocrine-disrupting effects, the industry is attempting to substitute BPA in variety of applications. Alternative phenolic compounds have been proposed for use in thermal paper; however, information to what extent BPA alternatives have been used in paper is sparse. The aim of the present work was to quantify BPA and its alternatives (bisphenol S (BPS), bisphenol E (BPE), bisphenol B (BPB), 4-cumylphenol (HPP) and bisphenol F (BPF)) in waste paper and board from Danish households, thermal paper receipts, non-carbon copy paper and conventional printer paper. BPA was found in all waste paper samples analysed, while BPS was identified in 73% of them. Only BPB was not identified in any of the samples. BPA and BPS were found in the majority of the receipts, which contained no measurable concentrations of the remaining alternatives. Although receipts showed the highest concentrations of BPA and BPS, office paper, flyers and corrugated boxes, together with receipts, represented the major flux of the two compounds in waste paper streams. Copyright © 2015 Elsevier Ltd. All rights reserved.

Inferring microbial interactions in thermophilic and mesophilic anaerobic digestion of hog waste.

PubMed

Shaw, Grace Tzun-Wen; Liu, An-Chi; Weng, Chieh-Yin; Chou, Chu-Yang; Wang, Daryi

2017-01-01

Anaerobic digestion (AnD) is a microbiological process that converts organic waste materials into biogas. Because of its high methane content, biogas is a combustible energy source and serves as an important environmental technology commonly used in the management of animal waste generated on large animal farms. Much work has been done on hardware design and process engineering for the generation of biogas. However, little is known about the complexity of the microbiology in this process. In particular, how microbes interact in the digester and eventually breakdown and convert organic matter into biogas is still regarded as a "black box." We used 16S rRNA sequencing as a tool to study the microbial community in laboratory hog waste digesters under tightly controlled conditions, and systematically unraveled the distinct interaction networks of two microbial communities from mesophilic (MAnD) and thermophilic anaerobic digestion (TAnD). Under thermophilic conditions, the well-known association between hydrogen-producing bacteria, e.g., Ruminococcaceae and Prevotellaceae, and hydrotrophic methanogens, Methanomicrobiaceae, was reverse engineered by their interactive topological niches. The inferred interaction network provides a sketch enabling the determination of microbial interactive relationships that conventional strategy of finding differential taxa was hard to achieve. This research is still in its infancy, but it can help to depict the dynamics of microbial ecosystems and to lay the groundwork for understanding how microorganisms cohabit in the anaerobic digester.

Evolution of microbial dynamics during the maturation phase of the composting of different types of waste.

PubMed

Villar, Iria; Alves, David; Garrido, Josefina; Mato, Salustiano

2016-08-01

During composting, facilities usually exert greater control over the bio-oxidative phase of the process, which uses a specific technology and generally has a fixed duration. After this phase, the material is deposited to mature, with less monitoring during the maturation phase. While there has been considerable study of biological parameters during the thermophilic phase, there is less research on the stabilization and maturation phase. This study evaluates the effects of the type of starting material on the evolution of microbial dynamics during the maturation phase of composting. Three waste types were used: sludge from the fish processing industry, municipal sewage sludge and pig manure, each independently mixed with shredded pine wood as bulking agent. The composting system for each waste type comprised a static reactor with capacity of 600L for the bio-oxidative phase followed by stabilization and maturation phase in triplicate 200L boxes for 112days. Phospholipid fatty acids, enzyme activities and physico-chemical parameters were measured throughout the maturation phase. The evolution of the total microbial biomass, Gram + bacteria, Gram - bacteria, fungi and enzymatic activities (β-glucosidase, cellulase, protease, acid and alkaline phosphatase) depended significantly on the waste type (p<0.001). The predominant microbial community for each waste type remained present throughout the maturation process, indicating that the waste type determines the microorganisms that are able to develop at this stage. While fungi predominated during fish sludge maturation, manure and municipal sludge were characterized by a greater proportion of bacteria. Both the structure of the microbial community and enzymatic activities provided important information for monitoring the composting process. More attention should be paid to the maturation phase in order to optimize composting. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Criticality assessment of TRU burial ground culverts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Winn, W.G.

1990-09-26

An effort to assess the criticality risks of {sup 239}Pu in TRU Burial Ground Culverts has been underway for several years. The concern arose from discrepancies in two types of monitors that have been used to assay the {sup 239}Pu waste prior to storage in 55-gallon drums that are placed in the culverts. One type is the solid waste monitor (SWM), which is based on gamma-ray measurements; the other is the neutron coincidence monitor, which is based on neutron measurements. The NCC was put into routine service after 1985 and has generally yielded higher 239 Pu assays than the SWM.more » Culverts with pre-1986 waste only had SWM assays of {sup 239}Pu; thus, it was questioned whether their actual {sup 239}Pu loadings could be high enough to pose criticality concerns. Studies to characterize the culvert criticality potential have included appraisal of NCC vs SWM, neutron measurements atop the culverts, gamma-ray measurements atop the culverts, and probabilistic risk analyses. Overall, these studies have implied that the culverts are critically safe; however, their results have not been examined collectively. The present report uses the collective information of the preceding studies to arrive at a more complete assessment of the culvert criticality aspects. A conservative k{sub eff} is estimated for an individual suspicious culvert and a PRA is evaluated for its {open_quotes}worst{close_quotes} drum. These two pieces of information form the basis of the appraisal, but other evidence is also included as support.« less

Evaluation of Faecalibacterium 16S rDNA genetic markers for accurate identification of swine faecal waste by quantitative PCR.

PubMed

Duan, Chuanren; Cui, Yamin; Zhao, Yi; Zhai, Jun; Zhang, Baoyun; Zhang, Kun; Sun, Da; Chen, Hang

2016-10-01

A genetic marker within the 16S rRNA gene of Faecalibacterium was identified for use in a quantitative PCR (qPCR) assay to detect swine faecal contamination in water. A total of 146,038 bacterial sequences were obtained using 454 pyrosequencing. By comparative bioinformatics analysis of Faecalibacterium sequences with those of numerous swine and other animal species, swine-specific Faecalibacterium 16S rRNA gene sequences were identified and Polymerase Chain Okabe (PCR) primer sets designed and tested against faecal DNA samples from swine and non-swine sources. Two PCR primer sets, PFB-1 and PFB-2, showed the highest specificity to swine faecal waste and had no cross-reaction with other animal samples. PFB-1 and PFB-2 amplified 16S rRNA gene sequences from 50 samples of swine with positive ratios of 86 and 90%, respectively. We compared swine-specific Faecalibacterium qPCR assays for the purpose of quantifying the newly identified markers. The quantification limits (LOQs) of PFB-1 and PFB-2 markers in environmental water were 6.5 and 2.9 copies per 100 ml, respectively. Of the swine-associated assays tested, PFB-2 was more sensitive in detecting the swine faecal waste and quantifying the microbial load. Furthermore, the microbial abundance and diversity of the microbiomes of swine and other animal faeces were estimated using operational taxonomic units (OTUs). The species specificity was demonstrated for the microbial populations present in various animal faeces. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Yeast Forkhead Transcription Factors Fkh1 and Fkh2 Regulate Lifespan and Stress Response Together with the Anaphase-Promoting Complex

PubMed Central

Postnikoff, Spike D. L.; Malo, Mackenzie E.; Wong, Berchman; Harkness, Troy A. A.

2012-01-01

Forkhead box O (FOXO) transcription factors have a conserved function in regulating metazoan lifespan. A key function in this process involves the regulation of the cell cycle and stress responses including free radical scavenging. We employed yeast chronological and replicative lifespan assays, as well as oxidative stress assays, to explore the potential evolutionary conservation of function between the FOXOs and the yeast forkhead box transcription factors FKH1 and FKH2. We report that the deletion of both FKH genes impedes normal lifespan and stress resistance, particularly in stationary phase cells, which are non-responsive to caloric restriction. Conversely, increased expression of the FKHs leads to extended lifespan and improved stress response. Here we show the Anaphase-Promoting Complex (APC) genetically interacts with the Fkh pathway, likely working in a linear pathway under normal conditions, as fkh1Δ fkh2Δ post-mitotic survival is epistatic to that observed in apc5CA mutants. However, under stress conditions, post-mitotic survival is dramatically impaired in apc5CA fkh1Δ fkh2Δ, while increased expression of either FKH rescues APC mutant growth defects. This study establishes the FKHs role as evolutionarily conserved regulators of lifespan in yeast and identifies the APC as a novel component of this mechanism under certain conditions, likely through combined regulation of stress response, genomic stability, and cell cycle regulation. PMID:22438832

Use of a Sentinel System for Field Measurements of Cryptosporidium parvum Oocyst Inactivation in Soil and Animal Waste

PubMed Central

Jenkins, M. B.; Walker, M. J.; Bowman, D. D.; Anthony, L. C.; Ghiorse, W. C.

1999-01-01

A small-volume sentinel chamber was developed to assess the effects of environmental stresses on survival of sucrose-Percoll-purified Cryptosporidium parvum oocysts in soil and animal wastes. Chambers were tested for their ability to equilibrate with external chemical and moisture conditions. Sentinel oocysts were then exposed to stresses of the external environment that affected their viability (potential infectivity), as indicated by results of a dye permeability assay. Preliminary laboratory experiments indicated that temperatures between 35 and 50°C and decreases in soil water potential (−0.003 to −3.20 MPa) increased oocyst inactivation rates. The effects of two common animal waste management practices on oocyst survival were investigated on three dairy farms in Delaware County, N.Y., within the New York City watershed: (i) piling wastes from dairy youngstock (including neonatal calves) and (ii) spreading wastes as a soil amendment on an agricultural field. Sentinel containers filled with air-dried and sieved (2-mm mesh) youngstock waste or field soil were wetted and inoculated with 2 million oocysts in an aqueous suspension and then placed in waste piles on two different farms and in soil within a cropped field on one farm. Controls consisted of purified oocysts in either phosphate-buffered saline or distilled water contained in sealed microcentrifuge tubes. Two microdata loggers recorded the ambient temperature at each field site. Sentinel experiments were conducted during the fall and winter (1996 to 1997) and winter (1998). Sentinel containers and controls were removed at 2- to 4-week intervals, and oocysts were extracted and tested by the dye permeability assay. The proportions of potentially infective oocysts exposed to the soil and waste pile material decreased more rapidly than their counterpart controls exposed to buffer or water, indicating that factors other than temperature affected oocyst inactivation in the waste piles and soil. The effect of soil freeze-thaw cycles was evident in the large proportion of empty sentinel oocysts. The potentially infective sentinel oocysts were reduced to <1% while the proportions in controls did not decrease below 50% potentially infective during the first field experiment. Microscopic observations of empty oocyst fragments indicated that abrasive effects of soil particles were a factor in oocyst inactivation. A similar pattern was observed in a second field experiment at the same site. PMID:10223991

Use of a sentinel system for field measurements of Cryptosporidium parvum oocyst inactivation in soil and animal waste.

PubMed

Jenkins, M B; Walker, M J; Bowman, D D; Anthony, L C; Ghiorse, W C

1999-05-01

A small-volume sentinel chamber was developed to assess the effects of environmental stresses on survival of sucrose-Percoll-purified Cryptosporidium parvum oocysts in soil and animal wastes. Chambers were tested for their ability to equilibrate with external chemical and moisture conditions. Sentinel oocysts were then exposed to stresses of the external environment that affected their viability (potential infectivity), as indicated by results of a dye permeability assay. Preliminary laboratory experiments indicated that temperatures between 35 and 50 degrees C and decreases in soil water potential (-0.003 to -3.20 MPa) increased oocyst inactivation rates. The effects of two common animal waste management practices on oocyst survival were investigated on three dairy farms in Delaware County, N.Y., within the New York City watershed: (i) piling wastes from dairy youngstock (including neonatal calves) and (ii) spreading wastes as a soil amendment on an agricultural field. Sentinel containers filled with air-dried and sieved (2-mm mesh) youngstock waste or field soil were wetted and inoculated with 2 million oocysts in an aqueous suspension and then placed in waste piles on two different farms and in soil within a cropped field on one farm. Controls consisted of purified oocysts in either phosphate-buffered saline or distilled water contained in sealed microcentrifuge tubes. Two microdata loggers recorded the ambient temperature at each field site. Sentinel experiments were conducted during the fall and winter (1996 to 1997) and winter (1998). Sentinel containers and controls were removed at 2- to 4-week intervals, and oocysts were extracted and tested by the dye permeability assay. The proportions of potentially infective oocysts exposed to the soil and waste pile material decreased more rapidly than their counterpart controls exposed to buffer or water, indicating that factors other than temperature affected oocyst inactivation in the waste piles and soil. The effect of soil freeze-thaw cycles was evident in the large proportion of empty sentinel oocysts. The potentially infective sentinel oocysts were reduced to <1% while the proportions in controls did not decrease below 50% potentially infective during the first field experiment. Microscopic observations of empty oocyst fragments indicated that abrasive effects of soil particles were a factor in oocyst inactivation. A similar pattern was observed in a second field experiment at the same site.

Functional elements in the minimal promoter of the human proton-coupled folate transporter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stark, Michal; Gonen, Nitzan; Assaraf, Yehuda G., E-mail: assaraf@tx.technion.ac.il

2009-10-09

The proton-coupled folate transporter (PCFT) is the dominant intestinal folate transporter, however, its promoter has yet to be revealed. Hence, we here cloned a 3.1 kb fragment upstream to the first ATG of the human PCFT gene and generated sequential deletion constructs evaluated in luciferase reporter assay. This analysis mapped the minimal promoter to 157 bp upstream to the first ATG. Crucial GC-box sites were identified within the minimal promoter and in its close vicinity which substantially contribute to promoter activity, as their disruption resulted in 94% loss of luciferase activity. We also identified upstream enhancer elements including YY1 andmore » AP1 which, although distantly located, prominently transactivated the minimal promoter, as their inactivation resulted in 50% decrease in reporter activity. This is the first functional identification of the minimal PCFT promoter harboring crucial GC-box elements that markedly contribute to its transcriptional activation via putative interaction with distal YY1 and AP1 enhancer elements.« less

Characterization of an RNA silencing suppressor encoded by maize yellow dwarf virus-RMV2.

PubMed

Wang, Fang; Zhao, Xia; Dong, Qing; Zhou, Benguo; Gao, Zhengliang

2018-05-11

Maize yellow dwarf virus-RMV2 (MYDV-RMV2) causes dwarfing and yellowing symptoms on leaves in field-grown maize plants in Anhui province in China. Herein, we evaluated the RNA silencing suppressor (RSS) activity of the P0 protein from MYDV-RMV2 by co-infiltration assays using wild-type and GFP-transgenic Nicotiana benthamiana (line 16C). The P0 of MYDV-RMV2 exhibited RSS activity and inhibited RNA silencing both locally and systemically. MYDV-RMV2 P0 acts as an F-box-like motif, and mutations to Ala at positions 67, 68, and 81 in the F-box-like motif (67LPxx81P) abolished the RSS activity of P0. However, MYDV-RMV2 P0 failed to interact with AGO1 from Arabidopsis thaliana. Expressing P0 induced developmental defects. P0 was targeted to both the nuclei and cytoplasm of plant cells. These findings expand our knowledge of the role of polerovirus P0 proteins in RNA silencing.

Pantoprazole blocks the JAK2/STAT3 pathway to alleviate skeletal muscle wasting in cancer cachexia by inhibiting inflammatory response

PubMed Central

Guo, Dunwei; Wang, Chaoyi; Wang, Qiang; Qiao, Zhongpeng; Tang, Hua

2017-01-01

Objective Cancer cachexia is often present in patients with advanced malignant tumors, and the subsequent body weight reduction results in poor quality of life. However, there has been no progress in developing effective clinical therapeutic strategies for skeletal muscle wasting in cancer cachexia. Herein, we explored the functions of pantoprazole on cancer cachexia skeletal muscle wasting. Methods The mouse colon adenocarcinoma cell line C26 was inoculated in the right forelimb of male BALB/C mice to establish a cancer cachexia model. The animals were treated with or without different concentrations of pantoprazole orally, and the body weight, tumor growth, spontaneous activity, and muscle functions were determined at various time points. Two weeks later, the levels of serum IL-6 and TNF-α, the mRNA levels of gastrocnemius JAK2 and STAT3, and the expression levels of p-JAK2, p-STAT3, Fbx32, and MuRF1 were examined with ELISA assay, qRT-PCR assay, and Western blotting, respectively. Further studies were performed to assess the levels of Fbx32 and MuRF1 expression and morphological changes. Results Pantoprazole can alleviate cancer cachexia-induced body weight reduction and inhibit skeletal muscle wasting in a dose-dependent manner. Our results indicated that pantoprazole treatment can decrease the levels of serum IL-6 and TNF-α (56.3% and 67.6%, respectively), and inhibit the activation of the JAK2/STAT3 signaling pathway. Moreover, the expression levels of MuRF1 and Fbx32 were also suppressed after pantoprazole treatment. Conclusion Our findings suggested that pantoprazole can alleviate cancer cachexia skeletal muscle wasting by inhibiting the inflammatory response and blocking the JAK2/STAT3 or ubiquitin proteasome pathway. PMID:28489606

Pantoprazole blocks the JAK2/STAT3 pathway to alleviate skeletal muscle wasting in cancer cachexia by inhibiting inflammatory response.

PubMed

Guo, Dunwei; Wang, Chaoyi; Wang, Qiang; Qiao, Zhongpeng; Tang, Hua

2017-06-13

Cancer cachexia is often present in patients with advanced malignant tumors, and the subsequent body weight reduction results in poor quality of life. However, there has been no progress in developing effective clinical therapeutic strategies for skeletal muscle wasting in cancer cachexia. Herein, we explored the functions of pantoprazole on cancer cachexia skeletal muscle wasting. The mouse colon adenocarcinoma cell line C26 was inoculated in the right forelimb of male BALB/C mice to establish a cancer cachexia model. The animals were treated with or without different concentrations of pantoprazole orally, and the body weight, tumor growth, spontaneous activity, and muscle functions were determined at various time points. Two weeks later, the levels of serum IL-6 and TNF-α, the mRNA levels of gastrocnemius JAK2 and STAT3, and the expression levels of p-JAK2, p-STAT3, Fbx32, and MuRF1 were examined with ELISA assay, qRT-PCR assay, and Western blotting, respectively. Further studies were performed to assess the levels of Fbx32 and MuRF1 expression and morphological changes. Pantoprazole can alleviate cancer cachexia-induced body weight reduction and inhibit skeletal muscle wasting in a dose-dependent manner. Our results indicated that pantoprazole treatment can decrease the levels of serum IL-6 and TNF-α (56.3% and 67.6%, respectively), and inhibit the activation of the JAK2/STAT3 signaling pathway. Moreover, the expression levels of MuRF1 and Fbx32 were also suppressed after pantoprazole treatment. Our findings suggested that pantoprazole can alleviate cancer cachexia skeletal muscle wasting by inhibiting the inflammatory response and blocking the JAK2/STAT3 or ubiquitin proteasome pathway.

Viral infection upregulates myostatin promoter activity in orange-spotted grouper (Epinephelus coioides)

PubMed Central

Chen, Yi-Tien; Lin, Chao-Fen; Chen, Young-Mao; Lo, Chih-En; Chen, Wan-Erh

2017-01-01

Myostatin is a negative regulator of myogenesis and has been suggested to be an important factor in the development of muscle wasting during viral infection. The objective of this study was to characterize the main regulatory element of the grouper myostatin promoter and to study changes in promoter activity due to viral stimulation. In vitro and in vivo experiments indicated that the E-box E6 is a positive cis-and trans-regulation motif, and an essential binding site for MyoD. In contrast, the E-box E5 is a dominant negative cis-regulatory. The characteristics of grouper myostatin promoter are similar in regulation of muscle growth to that of other species, but mainly through specific regulatory elements. According to these results, we conducted a study to investigate the effect of viral infection on myostatin promoter activity and its regulation. The nervous necrosis virus (NNV) treatment significantly induced myostatin promoter activity. The present study is the first report describing that specific myostatin motifs regulate promoter activity and response to viral infection. PMID:29036192

Viral infection upregulates myostatin promoter activity in orange-spotted grouper (Epinephelus coioides).

PubMed

Chen, Yi-Tien; Lin, Chao-Fen; Chen, Young-Mao; Lo, Chih-En; Chen, Wan-Erh; Chen, Tzong-Yueh

2017-01-01

Myostatin is a negative regulator of myogenesis and has been suggested to be an important factor in the development of muscle wasting during viral infection. The objective of this study was to characterize the main regulatory element of the grouper myostatin promoter and to study changes in promoter activity due to viral stimulation. In vitro and in vivo experiments indicated that the E-box E6 is a positive cis-and trans-regulation motif, and an essential binding site for MyoD. In contrast, the E-box E5 is a dominant negative cis-regulatory. The characteristics of grouper myostatin promoter are similar in regulation of muscle growth to that of other species, but mainly through specific regulatory elements. According to these results, we conducted a study to investigate the effect of viral infection on myostatin promoter activity and its regulation. The nervous necrosis virus (NNV) treatment significantly induced myostatin promoter activity. The present study is the first report describing that specific myostatin motifs regulate promoter activity and response to viral infection.

The mixed low-level waste problem in BE/NWN capsule

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hensley, D.C.

1999-07-01

The Boh Environmental, LLC (BE) and Northwest Nuclear, LLC (NWN) program addresses the problem of diminishing capacity in the United States to store mixed waste. A lack of an alternative program has caused the US Department of Energy (DOE) to indefinitely store all of its mixed waste in Resource Conservation and Recovery Act (RCRA) compliant storage facilities. Unfortunately, this capacity is fast approaching the administrative control limit. The combination of unique BE encapsulation and NWN waste characterization technologies provides an effective solution to DOE's mixed-waste dilemma. The BE ARROW-PAK technique encapsulates mixed low-level waste (MLLW) in extra-high molecular weight, high-densitymore » polyethylene, pipe-grade resin cylinders. ARROW-PAK applications include waste treatment, disposal, transportation (per 49 CFR 173), vault encasement, and interim/long-term storage for 100 to 300 yr. One of the first demonstrations of this treatment/storage technique successfully treated 880 mixed-waste debris drums at the DOE Hanford Site in 1997. NWN, deploying the APNea neutron assay technology, provides the screening and characterization capability necessary to ensure that radioactive waste is correctly categorized as either transuranic (TRU) or LLW. MLLW resulting from D and D activities conducted at the Oak Ridge East Tennessee Technology Park will be placed into ARROW-PAK containers following comprehensive characterization of the waste by NWN. The characterized and encapsulated waste will then be shipped to a commercial disposal facility, where the shipments meet all waste acceptance criteria of the disposal facility including treatment criteria.« less

Effect of liquid-to-solid ratio on semi-solid Fenton process in hazardous solid waste detoxication.

PubMed

Hu, Li-Fang; Feng, Hua-Jun; Long, Yu-Yang; Zheng, Yuan-Ge; Fang, Cheng-Ran; Shen, Dong-Sheng

2011-01-01

The liquid-to-solid ratio (L/S) of semi-solid Fenton process (SSFP) designated for hazardous solid waste detoxication was investigated. The removal and minimization effects of o-nitroaniline (ONA) in simulate solid waste residue (SSWR) from organic arsenic industry was evaluated by total organic carbon (TOC) and ONA removal efficiency, respectively. Initially, Box-Behnken design (BBD) and response surface methodology (RSM) were used to optimize the key factors of SSFP. Results showed that the removal rates of TOC and ONA decreased as L/S increased. Subsequently, four target initial ONA concentrations including 100 mg kg(-1), 1 g kg(-1), 10 g kg(-1), and 100 gk g(-1) on a dry basis were evaluated for the effect of L/S. A significant cubic empirical model between the initial ONA concentration and L/S was successfully developed to predict the optimal L/S for given initial ONA concentration for SSFP. Moreover, an optimized operation strategy of multi-SSFP for different cases was determined based on the residual target pollutant concentration and the corresponding environmental conditions. It showed that the total L/S of multi-SSFP in all tested scenarios was no greater than 3.8, which is lower than the conventional slurry systems (L/S ≥ 5). The multi-SSFP is environment-friendly when it used for detoxication of hazardous solid waste contaminated by ONA and provides a potential method for the detoxication of hazardous solid waste contaminated by organics. Copyright © 2010 Elsevier Ltd. All rights reserved.

Anaerobic co-digestion of commercial food waste and dairy manure: Characterizing biochemical parameters and synergistic effects.

PubMed

Ebner, Jacqueline H; Labatut, Rodrigo A; Lodge, Jeffrey S; Williamson, Anahita A; Trabold, Thomas A

2016-06-01

Anaerobic digestion of commercial food waste is a promising alternative to landfilling commercial food waste. This study characterized 11 types of commercial food wastes and 12 co-digestion blends. Bio-methane potential, biodegradable fraction, and apparent first-order hydrolysis rate coefficients were reported based upon biochemical methane potential (BMP) assays. Food waste bio-methane potentials ranged from 165 to 496 mL CH4/g VS. Substrates high in lipids or readily degradable carbohydrates showed the highest methane production. Average bio-methane potential observed for co-digested substrates was -5% to +20% that of the bio-methane potential of the individual substrates weighted by VS content. Apparent hydrolysis rate coefficients ranged from 0.19d(-1) to 0.65d(-1). Co-digested substrates showed an accelerated apparent hydrolysis rate relative to the weighted average of individual substrate rates. These results provide a database of key bio-digestion parameters to advance modeling and utilization of commercial food waste in anaerobic digestion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bread enriched in lycopene and other bioactive compounds by addition of dry tomato waste.

PubMed

Nour, Violeta; Ionica, Mira Elena; Trandafir, Ion

2015-12-01

The tomato processing industry generates high amounts of waste, mainly tomato skins and seeds, which create environmental problems. These residues are attractive sources of valuable bioactive components and pigments. A relatively simple recovery technology could consist of production of powders to be directly incorporated into foods. Tomato waste coming from a Romanian tomato processing unit were analyzed for the content of several bioactive compounds like ascorbic acid, β-carotene, lycopene, total phenolics, mineral and trace elements. In addition, its antioxidant capacity was assayed. Results revealed that tomato waste (skins and seeds) could be successfully utilized as functional ingredient for the formulation of antioxidant rich functional foods. Dry tomato processing waste were used to supplement wheat flour at 6 and 10 % levels (w/w flour basis) and the effects on the bread's physicochemical, baking and sensorial characteristics were studied. The following changes were observed: increase in moisture content, titratable acidity and bread crumb elasticity, reduction in specific volume and bread crumb porosity. The addition of dry tomato waste at 6 % resulted in bread with good sensory characteristics and overall acceptability but as the amount of dry tomato waste increased to 10 %, bread was less acceptable.

Two Virus Based Endocrine Disruptor Assays Effective Across Vertebrate Classes.

EPA Science Inventory

The presence of hormone mimics, or endocrine disrupting compounds (EDC’s), in the environment are increasing. Sources range from agricultural run–off, pharmaceuticals in waste water, to industrial operations. Current levels of contamination are sufficient to alter sexual develo...

Quantitative CrAssphage PCR Assays for Human Fecal Pollution Measurement

EPA Science Inventory

Environmental waters are monitored for fecal pollution to protect public health and water resources. Traditionally, general fecal indicator bacteria are used; however, they cannot distinguish human fecal waste from pollution from other animals. Recently, a novel bacteriophage, cr...

The SBP-Box Gene VpSBP11 from Chinese Wild Vitis Is Involved in Floral Transition and Affects Leaf Development.

PubMed

Hou, Hongmin; Yan, Xiaoxiao; Sha, Ting; Yan, Qin; Wang, Xiping

2017-07-13

Flowering occurs in angiosperms during a major developmental transition from vegetative growth to the reproductive phase. Squamosa promoter binding protein (SBP)-box genes have been found to play critical roles in regulating flower and fruit development, but their roles in grapevine have remained unclear. To better understand the functions of the grape SBP-box genes in both vegetative and reproductive growth phases, a full-length complementary DNA (cDNA) sequence of the putative SBP-box transcription factor gene, VpSBP11 , was obtained from Chinese wild grapevine Vitis pseudoreticulata Wen Tsai Wang (W. T. Wang) clone 'Baihe-35-1'. VpSBP11 encoded a putative polypeptide of 170 amino acids with a highly conserved SBP-domain with two zinc-binding sites of the Cx2C-x3-H-x11-C-x6-H (C2HCH) type and a nuclear localization signal. We confirmed that the VpSBP11 protein was targeted to the nucleus and possessed transcriptional activation activity by subcellular localization and trans -activation assay. Over-expression of VpSBP11 in Arabidopsis thaliana was shown to activate the FUL gene, and subsequently the AP1 and LFY genes, all of which were floral meristem identity genes, and to cause earlier flowering than in wild type (WT) plants. The pattern of vegetative growth was also different between the transgenic and WT plants. For example, in the VpSBP11 over-expressing transgenic plants, the number of rosette leaves was less than that of WT; the petiole was significantly elongated; and the rosette and cauline leaves curled upwards or downwards. These results were consistent with VpSBP11 acting as a transcription factor during the transition from the vegetative stage to the reproductive stage.

A human Polycomb isoform lacking the Pc box does not participate to PRC1 complexes but forms protein assemblies and represses transcription.

PubMed

Völkel, Pamela; Le Faou, Perrine; Vandamme, Julien; Pira, Dorcas; Angrand, Pierre-Olivier

2012-05-01

Polycomb repression controls the expression of hundreds of genes involved in development and is mediated by essentially two classes of chromatin-associated protein complexes. The Polycomb repressive complex 2 (PRC2) trimethylates histone H3 at lysine 27, an epigenetic mark that serves as a docking site for the PRC1 protein complex. Drosophila core PRC1 is composed of four subunits: Polycomb (Pc), Posterior sex combs (Psc), Polyhomeotic (Ph) and Sex combs extra (Sce). Each of these proteins has multiple orthologs in vertebrates, thus generating an enormous scope for potential combinatorial diversity. In particular, mammalian genomes encode five Pc family members: CBX2, CBX4, CBX6, CBX7 and CBX8. To complicate matters further, distinct isoforms might arise from single genes. Here, we address the functional role of the two human CBX2 isoforms. Owing to different polyadenylation sites and alternative splicing events, the human CBX2 locus produces two transcripts: a 5-exon transcript that encodes the 532-amino acid CBX2-1 isoform that contains the conserved chromodomain and Pc box and a 4-exon transcript encoding a shorter isoform, CBX2-2, lacking the Pc box but still possessing a chromodomain. Using biochemical approaches and a novel in vivo imaging assay, we show that the short CBX2-2 isoform lacking the Pc box, does not participate in PRC1 protein complexes, but self-associates in vivo and forms complexes of high molecular weight. Furthermore, the CBX2 short isoform is still able to repress transcription, suggesting that Polycomb repression might occur in the absence of PRC1 formation.

Terrestrial predator alarm vocalizations are a valid monitor of stress in captive brown capuchins (Cebus apella)

USGS Publications Warehouse

Boinski, S.; Gross, T.S.; Davis, J.K.

1999-01-01

The vocal behavior of captive animals is increasingly exploited as an index of well-being. Here we show that the terrestrial predator alarm (TPA) vocalization, a robust and acoustically distinctive anti-predation vocal response present in many mammal and bird species, offers useful information on the relative well-being and stress levels of captive animals. In a 16-week experiment evaluating the effects of varying levels of physical environmental enrichment (control < toys < foraging box < foraging box and toys) in the cages of eight singly housed adult male brown capuchins, we quantified the 1) emission rate of TPAs, 2) proportions of normal and abnormal behavior sample intervals, and 3) fecal and plasma cortisol levels. Variation in TPA emission across the experimental conditions was significant. We found significant reductions in the mean TPA production rate by the group in the enriched (toys, foraging box, and foraging box and toys) compared to the control condition; pre-and post-experimental conditions, however, did not differ from the control condition. Mean TPA production by the group was also significantly positively correlated to mean group levels of fecal cortisol and proportion of abnormal behavior sample intervals, and significantly negatively correlated to the average proportion of normal behavior sample intervals in the group. Based on group means, plasma cortisol levels were positively, but not significantly, related to increasing TPA rate. At the level of the responses of an individual subject, however, the covariation between the vocal and non-vocal behavioral measures and the cortisol assays seldom attained significance. Nevertheless, the direction of the relationships among these parameters within individual subjects typically mirrored those correlations based on group means. At both the group mean and individual levels, our results are consistent with the.

Suppression of grp78 core promoter element-mediated stress induction by the dbpA and dbpB (YB-1) cold shock domain proteins.

PubMed Central

Li, W W; Hsiung, Y; Wong, V; Galvin, K; Zhou, Y; Shi, Y; Lee, A S

1997-01-01

The highly conserved grp78 core promoter element plays an important role in the induction of grp78 under diverse stress signals. Previous studies have established a functional region in the 3' half of the core (stress-inducible change region [SICR]) which exhibits stress-inducible changes in stressed nuclei. The human transcription factor YY1 is shown to bind the SICR and transactivate the core element under stress conditions. Here we report that expression library screening with the core element has identified two new core binding proteins, YB-1 and dbpA. Both proteins belong to the Y-box family of proteins characterized by an evolutionarily conserved DNA binding motif, the cold shock domain (CSD). In contrast to YY1, which binds only double-stranded SICR, the Y-box/CSD proteins much prefer the lower strand of the SICR. The Y-box proteins can repress the inducibility of the grp78 core element mediated by treatment of cells with A23187, thapsigargin, and tunicamycin. In gel shift assays, YY1 binding to the core element is inhibited by either YB-1 or dbpA. A yeast interaction trap screen using LexA-YY1 as a bait and a HeLa cell cDNA-acid patch fusion library identified YB-1 as a YY1-interacting protein. In cotransfection experiments, the Y-box proteins antagonize the YY1-mediated enhancement of transcription directed by the grp78 core in stressed cells. Thus, the CSD proteins may be part of the stress signal transduction mechanism in the mammalian system. PMID:8972186

SMA-Causing Missense Mutations in Survival motor neuron (Smn) Display a Wide Range of Phenotypes When Modeled in Drosophila

PubMed Central

Praveen, Kavita; Wen, Ying; Gray, Kelsey M.; Noto, John J.; Patlolla, Akash R.; Van Duyne, Gregory D.; Matera, A. Gregory

2014-01-01

Mutations in the human survival motor neuron 1 (SMN) gene are the primary cause of spinal muscular atrophy (SMA), a devastating neuromuscular disorder. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. Additional tissue-specific and global functions have been ascribed to SMN; however, their relevance to SMA pathology is poorly understood and controversial. Using Drosophila as a model system, we created an allelic series of twelve Smn missense mutations, originally identified in human SMA patients. We show that animals expressing these SMA-causing mutations display a broad range of phenotypic severities, similar to the human disease. Furthermore, specific interactions with other proteins known to be important for SMN's role in RNP assembly are conserved. Intragenic complementation analyses revealed that the three most severe mutations, all of which map to the YG box self-oligomerization domain of SMN, display a stronger phenotype than the null allele and behave in a dominant fashion. In support of this finding, the severe YG box mutants are defective in self-interaction assays, yet maintain their ability to heterodimerize with wild-type SMN. When expressed at high levels, wild-type SMN is able to suppress the activity of the mutant protein. These results suggest that certain SMN mutants can sequester the wild-type protein into inactive complexes. Molecular modeling of the SMN YG box dimer provides a structural basis for this dominant phenotype. These data demonstrate that important structural and functional features of the SMN YG box are conserved between vertebrates and invertebrates, emphasizing the importance of self-interaction to the proper functioning of SMN. PMID:25144193

SMA-causing missense mutations in survival motor neuron (Smn) display a wide range of phenotypes when modeled in Drosophila.

PubMed

Praveen, Kavita; Wen, Ying; Gray, Kelsey M; Noto, John J; Patlolla, Akash R; Van Duyne, Gregory D; Matera, A Gregory

2014-08-01

Mutations in the human survival motor neuron 1 (SMN) gene are the primary cause of spinal muscular atrophy (SMA), a devastating neuromuscular disorder. SMN protein has a well-characterized role in the biogenesis of small nuclear ribonucleoproteins (snRNPs), core components of the spliceosome. Additional tissue-specific and global functions have been ascribed to SMN; however, their relevance to SMA pathology is poorly understood and controversial. Using Drosophila as a model system, we created an allelic series of twelve Smn missense mutations, originally identified in human SMA patients. We show that animals expressing these SMA-causing mutations display a broad range of phenotypic severities, similar to the human disease. Furthermore, specific interactions with other proteins known to be important for SMN's role in RNP assembly are conserved. Intragenic complementation analyses revealed that the three most severe mutations, all of which map to the YG box self-oligomerization domain of SMN, display a stronger phenotype than the null allele and behave in a dominant fashion. In support of this finding, the severe YG box mutants are defective in self-interaction assays, yet maintain their ability to heterodimerize with wild-type SMN. When expressed at high levels, wild-type SMN is able to suppress the activity of the mutant protein. These results suggest that certain SMN mutants can sequester the wild-type protein into inactive complexes. Molecular modeling of the SMN YG box dimer provides a structural basis for this dominant phenotype. These data demonstrate that important structural and functional features of the SMN YG box are conserved between vertebrates and invertebrates, emphasizing the importance of self-interaction to the proper functioning of SMN.

Development and application of a sensitive, phenotypic, high-throughput image-based assay to identify compound activity against Trypanosoma cruzi amastigotes

PubMed Central

Sykes, Melissa L.; Avery, Vicky M.

2015-01-01

We have developed a high content 384-well, image-based assay to estimate the effect of compound treatment on Trypanosoma cruzi amastigotes in 3T3 fibroblasts. In the same well, the effect of compound activity on host cells can also be determined, as an initial indicator of cytotoxicity. This assay has been used to identify active compounds from an in-house library of compounds with either known biological activity or that are FDA approved, and separately, from the Medicines for Malaria Venture Malaria Box collection. Active compounds were screened against T. cruzi trypomastigotes, utilising an assay developed with the viability dye resazurin. Twelve compounds with reconfirmed solid sample activity, with IC50 values of less than 10 μM and selectivity indices to T. cruzi amastigotes over 3T3 host cells of between >22 and 319 times were identified from these libraries. As 3T3 cells are contact inhibited, with limited proliferation in the assay, selective compounds of interest were profiled in a separate assay to estimate the viability of compound treated, replicating HEK293 cells. Selective compounds that were not previously reported in the literature were further profiled by extending the incubation time against amastigote infected 3T3 cells to determine if there were residual amastigotes post-treatment, important for the consideration of the exposure time required for further biological characterisation. The assay development process and the suitability of identified compounds as hit molecules for Chagas disease research are discussed. PMID:27120069

Development and application of a sensitive, phenotypic, high-throughput image-based assay to identify compound activity against Trypanosoma cruzi amastigotes.

PubMed

Sykes, Melissa L; Avery, Vicky M

2015-12-01

We have developed a high content 384-well, image-based assay to estimate the effect of compound treatment on Trypanosoma cruzi amastigotes in 3T3 fibroblasts. In the same well, the effect of compound activity on host cells can also be determined, as an initial indicator of cytotoxicity. This assay has been used to identify active compounds from an in-house library of compounds with either known biological activity or that are FDA approved, and separately, from the Medicines for Malaria Venture Malaria Box collection. Active compounds were screened against T. cruzi trypomastigotes, utilising an assay developed with the viability dye resazurin. Twelve compounds with reconfirmed solid sample activity, with IC50 values of less than 10 μM and selectivity indices to T. cruzi amastigotes over 3T3 host cells of between >22 and 319 times were identified from these libraries. As 3T3 cells are contact inhibited, with limited proliferation in the assay, selective compounds of interest were profiled in a separate assay to estimate the viability of compound treated, replicating HEK293 cells. Selective compounds that were not previously reported in the literature were further profiled by extending the incubation time against amastigote infected 3T3 cells to determine if there were residual amastigotes post-treatment, important for the consideration of the exposure time required for further biological characterisation. The assay development process and the suitability of identified compounds as hit molecules for Chagas disease research are discussed. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Chips in black boxes? Convenience life span, parafood, brandwidth, families, and co-creation.

PubMed

Jacobs, Marc

2015-11-01

Any consumer who opens a bag of potato or corn chips (or crisps in the UK) knows there is no time to waste to enjoy or share them. The convenience life span of chips is limited: it is the shelf or storage life and a very limited time once outside the bag. Many technologies converge to generate the desired effect as a black box, not only of the packaging but also of the chips themselves. The concept of paratext can be applied to printed messages on the package, including the brand name and other texts like advertising (epitexts), which can be expanded into the concept of parafood. These concepts help to discuss technological developments and interpret why this has recently become a negotiation zone for co-creation (see the Do us a flavor campaigns). They are symptoms of changing relations between production, research and development, marketing, and consumption. This paper pays special attention to back stories, underdog brand biographies and narratives about origin. The concept of brandwidth is introduced to sensitize about the limits of combining different stories about chips. A recent brand biography, a family history and a cookery book are used to discuss the phenomenon of cooking with Fritos. Together with the concepts of parafood, brandwidth and black boxes, more reflection and dialogue about the role of history and heritage in marketing put new challenging perspectives on the agenda. Copyright © 2015 Elsevier Ltd. All rights reserved.

Curcumin induces apoptosis in pancreatic cancer cells through the induction of forkhead box O1 and inhibition of the PI3K/Akt pathway.

PubMed

Zhao, Zhiming; Li, Chenggang; Xi, Hao; Gao, Yuanxing; Xu, Dabin

2015-10-01

Previous population investigations have suggested that the application of curcumin may be associated with decreased incidence and improved prognosis in certain types of cancer. Forkhead box O1 (FOXO1) has been implicated in the regulation of several biological processes, including stress resistance, metabolism, DNA repair, cell cycle and apoptosis. The aims of the present study were to investigate the effects and molecular mechanisms of curcumin on the induction of anti‑proliferation, cell cycle arrest and apoptosis, by FOXO1, in pancreatic cancer cells. The MTT assay and ELISA‑Brdu assay were used to assess cell proliferation. Reverse transcription‑quantitative polymerase chain reaction and western blot analyses were used to detect the expression of PCNA, Ki‑67, B‑cell lymphoma‑2 (Bcl‑2), B‑cell‑associated X protein (Bax), cyclin D1, p21, p27 and FOXO1. Cell apoptosis was detected using a Cell Death ELISA detection kit. A Caspase‑3/9 Fluorescent Assay kit was used to detect caspase activity. The findings revealed that curcumin significantly decreased cell proliferation, which was associated with increased expression of the p21/CIP1 and p27/KIP1 cyclin‑dependent kinase inhibitors, and inhibited expression of cyclin D1. In addition, curcumin induced apoptosis by decreasing the Bcl‑2/Bax protein ratio and increasing caspase‑9/3 activation in the pancreatic cancer cells. Using siRNA against FOXO1, and Akt inhibitor and activator, the present study confirmed that curcumin induced the expression of FOXO1 by inhibition of phosphoinositide 3‑kinase/Akt signaling, leading to cell cycle arrest and apoptosis. In conclusion, these findings offer support for a mechanism that may underlie the anti‑neoplastic effects of curcumin and justify further investigation to examine the potential roles for activators of FOXO1 in the prevention and treatment of pancreatic cancer.

Dealing with Historical Discrepancies: The Recovery of National Research Experiment (NRX) Reactor Fuel Rods at Chalk River Laboratories (CRL) - 13324

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vickerd, Meggan

2013-07-01

Following the 1952 National Research Experiment (NRX) Reactor accident, fuel rods which had short irradiation histories were 'temporarily' buried in wooden boxes at the 'disposal grounds' during the cleanup effort. The Nuclear Legacy Liabilities Program (NLLP), funded by Natural Resources Canada (NRCan), strategically retrieves legacy waste and restores lands affected by Atomic Energy of Canada Limited (AECL) early operations. Thus under this program the recovery of still buried NRX reactor fuel rods and their relocation to modern fuel storage was identified as a priority. A suspect inventory of NRX fuels was compiled from historical records and various research activities. Sitemore » characterization in 2005 verified the physical location of the fuel rods and determined the wooden boxes they were buried in had degraded such that the fuel rods were in direct contact with the soil. The fuel rods were recovered and transferred to a modern fuel storage facility in 2007. Recovered identification tags and measured radiation fields were used to identify the inventory of these fuels. During the retrieval activity, a discrepancy was discovered between the anticipated number of fuel rods and the number found during the retrieval. A total of 32 fuel rods and cans of cut end pieces were recovered from the specified site, which was greater than the anticipated 19 fuel rods and cans. This discovery delayed the completion of the project, increased the associated costs, and required more than anticipated storage space in the modern fuel storage facility. A number of lessons learned were identified following completion of this project, the most significant of which was the potential for discrepancies within the historical records. Historical discrepancies are more likely to be resolved by comprehensive historical record searches and site characterizations. It was also recommended that a complete review of the wastes generated, and the total affected lands as a result of this historic 1952 NRX accident be undertaken. These lessons and recommendations have lead to changes in how the NLLP is executed in the CRL waste management areas. (authors)« less

Evaluation of an Experimental Re-introduction of Sockeye Salmon into Skaha Lake; Year 3 of 3; Addendum to the Assessment of Juvenile Oncorhynchus nerka (Sockeye and Kokane) Rearing Conditions of Skaha and Osoyoos Lakes 2002 Section of the 2002 Technical Report, 2003 Technical Report.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wright, Howie; Lawrence, Shayla; Rebellato, Betty

2004-01-01

The purpose of this addendum is, first, to provide and discuss disease agent survey results that were not available for inclusion in the Disease Risk Assessment portion of the YEAR 3 report at the time of its writing, and second, to make recommendations stemming from these results. The first set of results deals with live box exposure tests conducted using juvenile sentinel rainbow trout in the spring of 2002 to detect Myxosoma cerebralis and Ceratomyxa shasta. The second set of results deals with similar exposure tests conducted in the spring of 2003. The latter tests were initially intended to occurmore » in the fall of 2002 but had to be re-scheduled to the spring of 2003 because suitably aged sentinel rainbow trout for the exposures were not available in the fall of 2002. The methods used for the live box exposure tests were essentially the same as those described in the YEAR 3 report. Fish were again exposed at the same four sites above McIntyre Dam and at the same four sites below the dam. As mentioned in the YEAR 3 report, the spring 2002 exposure lasted for 21 days (May 6 to 27). The spring 2003 exposure also lasted for 21 days (April 22 to May 13). The number of fish in the spring 2003 tests was, however, reduced to approximately half the number used in previous tests in order to reduce the chances of dissolved oxygen problems, suspected to have occurred in earlier tests in some of the live boxes. As before, fish that survived the live box exposures were transferred to Skaha Hatchery where they were held for sufficient time to permit any infections with M. cerebralis and C. shasta to develop and to permit for spore development in these pathogens. Assays for the pathogens were carried out as previously described. Detection of M. cerebralis was based on detecting its spores following the trypsin/pepsin digestion method. Detection of C. shasta was based on a polymerase chain reaction (PCR) test, but smears of fresh intestinal tissues (one fish per smear) were also prepared so that positive PCR findings could be confirmed by the microscopic observation of C. shasta spores. Except as just mentioned, appropriate tissues from the fish were in most cases pooled (maximum of five fish per pool) for the assays.« less

Establishing a safe, rapid, convenient and low-cost antiviral assay of interferon bioactivity based on recombinant VSV expressing GFP.

PubMed

Chen, Weiye; Wen, Zhiyuan; Zhang, Jialin; Li, Cuicui; Huang, Kehe; Bu, Zhigao

2018-02-01

The methods of the quantitative assay of the antiviral activity of interferons (IFNs) (type I, II or III) are very important during carrying out of the research of them, since they were found. Here a recombinant vesicular stomatitis virus expressing green fluorescent protein (GFP) (VSV/GFP) and MDBK cells were used to develop an antiviral assay (AVA) for IFNs. This method was carried out on a 96-well cell culture plate, and the half reduction of virus replication was quantified by assaying GFP. To quantify GFP, cell lysis buffer was directly added to the wells infected with VSV/GFP to lyse cells, the VSV/GFP was then inactivated, and relative fluorescence unit (RFU) of GFP was measured and used to calculate the antiviral activity. This method needed only one step instead of three steps in the staining method with naphthol blue black, medium with phenol red can be used, and it had good reproducibility. The GFP-containing samples could be stored at 4°C in a wet box for at least 1 week without affecting the assay results. In addition, the results obtained with this method were similar to those obtained with the staining method. In conclusion, a safe, rapid, convenient and low-cost AVA of IFN based on recombinant VSV/GFP was established. Copyright © 2017. Published by Elsevier B.V.

Cloning and characterization of the mouse XPAC gene.

PubMed Central

van Oostrom, C T; de Vries, A; Verbeek, S J; van Kreijl, C F; van Steeg, H

1994-01-01

Xeroderma Pigmentosum is a human disease, which is, among others, characterized by a high incidence of (sunlight induced) skin cancer, due to a defect in nucleotide excision repair (NER). The human DNA repair gene XPAC corrects this defect in cells isolated from Xeroderma Pigmentosum complementation group A (XP-A) patients. To enable the development of a transgenic mouse model for XP-A by gene targeting in embryonic stem cells, we cloned and characterized the mouse homologue of the XPAC gene. The mouse XPAC gene was found to consist of 6 exons, spanning approximately 21 kb. The nucleotide sequence of the exons is identical to that of the also cloned the mouse XPAC cDNA. Furthermore, the deduced amino acid sequence of the XPAC protein is the same as the one published previously by Tanaka et al. From CAT assay analysis, the promoter of the XPAC gene appeared to be located within 313 bp upstream of the assumed transcriptional start site. Like the promoters of other eukaryotic DNA repair genes (i.e. ERCC-1 and XPBC/ERCC-3), the mouse XPAC promoter region lacks classical promoter elements like TATA-, GC- and CAAT boxes. However, it contains an unique polypyrimidine-rich box, which is so far only found in genes encoding DNA repair enzymes. The function of this box in the regulation of transcription is still unclear. PMID:8127648

Functional characterization of GhSOC1 and GhMADS42 homologs from upland cotton (Gossypium hirsutum L.).

PubMed

Zhang, Xiaohong; Wei, Jianghui; Fan, Shuli; Song, Meizhen; Pang, Chaoyou; Wei, Hengling; Wang, Chengshe; Yu, Shuxun

2016-01-01

In Arabidopsis flowering pathway, MADS-box genes encode transcription factors, with their structures and functions highly conserved in many species. In our study, two MADS-box genes GhSOC1 and GhMADS42 (Gossypium hirsutum L.) were cloned from upland cotton CCRI36 and transformed into Arabidopsis. GhSOC1 was additionally transformed into upland cotton. Comparative analysis demonstrated sequence conservation between GhSOC1 and GhMADS42 and genes of other plant species. Tissue-specific expression analysis of GhSOC1 and GhMADS42 revealed spatiotemporal expression patterns involving high transcript levels in leaves, shoot apical buds, and flowers. In addition, overexpression of both GhSOC1 and GhMADS42 in Arabidopsis accelerated flowering, with GhMADS42 transgenic plants showing abnormal floral organ phenotypes. Overexpression of GhSOC1 in upland cotton also produced variations in floral organs. Furthermore, chromatin immunoprecipitation assay demonstrated that GhSOC1 could regulate GhMADS41 and GhMADS42, but not FLOWERING LOCUS T, by directly binding to the genes promoter. Finally, yeast two-hybrid and bimolecular fluorescence complementation approaches were undertaken to better understand the interaction of GhSOC1 and other MADS-box factors. These experiments showed that GhSOC1 can interact with APETALA1/FRUITFULL-like proteins in cotton. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Scleraxis is a transcriptional activator that regulates the expression of Tenomodulin, a marker of mature tenocytes and ligamentocytes.

PubMed

Shukunami, Chisa; Takimoto, Aki; Nishizaki, Yuriko; Yoshimoto, Yuki; Tanaka, Seima; Miura, Shigenori; Watanabe, Hitomi; Sakuma, Tetsushi; Yamamoto, Takashi; Kondoh, Gen; Hiraki, Yuji

2018-02-16

Tenomodulin (Tnmd) is a type II transmembrane glycoprotein predominantly expressed in tendons and ligaments. We found that scleraxis (Scx), a member of the Twist-family of basic helix-loop-helix transcription factors, is a transcriptional activator of Tnmd expression in tenocytes. During embryonic development, Scx expression preceded that of Tnmd. Tnmd expression was nearly absent in tendons and ligaments of Scx-deficient mice generated by transcription activator-like effector nucleases-mediated gene disruption. Tnmd mRNA levels were dramatically decreased during serial passages of rat tenocytes. Scx silencing by small interfering RNA significantly suppressed endogenous Tnmd mRNA levels in tenocytes. Mouse Tnmd contains five E-box sites in the ~1-kb 5'-flanking region. A 174-base pair genomic fragment containing a TATA box drives transcription in tenocytes. Enhancer activity was increased in the upstream region (-1030 to -295) of Tnmd in tenocytes, but not in NIH3T3 and C3H10T1/2 cells. Preferential binding of both Scx and Twist1 as a heterodimer with E12 or E47 to CAGATG or CATCTG and transactivation of the 5'-flanking region were confirmed by electrophoresis mobility shift and dual luciferase assays, respectively. Scx directly transactivates Tnmd via these E-boxes to positively regulate tenocyte differentiation and maturation.

Corrective Action Decision Document for Corrective Action Unit 428: Area 3 Septic Waste Systems 1 and 5, Tonopah Test Range, Nevada

DOE Office of Scientific and Technical Information (OSTI.GOV)

U.S. Department of Energy, Nevada Operations Office

2000-02-08

This Corrective Action Decision Document identifies and rationalizes the US Department of Energy, Nevada Operations Office's selection of a recommended corrective action alternative (CAA) appropriate to facilitate the closure of Corrective Action Unit (CAU) 428, Septic Waste Systems 1 and 5, under the Federal Facility Agreement and Consent Order. Located in Area 3 at the Tonopah Test Range (TTR) in Nevada, CAU 428 is comprised of two Corrective Action Sites (CASs): (1) CAS 03-05-002-SW01, Septic Waste System 1 and (2) CAS 03-05-002- SW05, Septic Waste System 5. A corrective action investigation performed in 1999 detected analyte concentrations that exceeded preliminarymore » action levels; specifically, contaminants of concern (COCs) included benzo(a) pyrene in a septic tank integrity sample associated with Septic Tank 33-1A of Septic Waste System 1, and arsenic in a soil sample associated with Septic Waste System 5. During this investigation, three Corrective Action Objectives (CAOs) were identified to prevent or mitigate exposure to contents of the septic tanks and distribution box, to subsurface soil containing COCs, and the spread of COCs beyond the CAU. Based on these CAOs, a review of existing data, future use, and current operations in Area 3 of the TTR, three CAAs were developed for consideration: Alternative 1 - No Further Action; Alternative 2 - Closure in Place with Administrative Controls; and Alternative 3 - Clean Closure by Excavation and Disposal. These alternatives were evaluated based on four general corrective action standards and five remedy selection decision factors. Based on the results of the evaluation, the preferred CAA was Alternative 3. This alternative meets all applicable state and federal regulations for closure of the site and will eliminate potential future exposure pathways to the contaminated soils at the Area 3 Septic Waste Systems 1 and 5.« less

Optimization of magnetic field-assisted ultrasonication for the disintegration of waste activated sludge using Box-Behnken design with response surface methodology.

PubMed

Guan, Su; Deng, Feng; Huang, Si-Qi; Liu, Shu-Yang; Ai, Le-Xian; She, Pu-Ying

2017-09-01

This study investigated for the first time the feasibility of using a magnetic field for sludge disintegration. Approximately 41.01% disintegration degree (DD) was reached after 30min at 180mT magnetic field intensity upon separate magnetic field treatment. Protein and polysaccharide contents significantly increased. This test was optimized using a Box-Behnken design (BBD) with response surface methodology (RSM) to fit the multiple equation of the DD. The maximum DD was 43.75% and the protein and polysaccharide contents increased to 56.71 and 119.44mg/L, respectively, when the magnetic field strength was 119.69mT, reaction time was 30.49min, and pH was 9.82 in the optimization experiment. We then analyzed the effects of ultrasound alone. We are the first to combine magnetic field with ultrasound to disintegrate waste-activated sludge (WAS). The optimum effect was obtained with the application of ultrasound alone at 45kHz frequency, with a DD of about 58.09%. By contrast, 62.62% DD was reached in combined magnetic field and ultrasound treatment. This combined test was also optimized using BBD with RSM to fit the multiple equation of DD. The maximum DD of 64.59% was achieved when the magnetic field intensity was 197.87mT, ultrasonic frequency was 42.28kHz, reaction time was 33.96min, and pH was 8.90. These results were consistent with those of particle size and electron microscopy analyses. This research proved that a magnetic field can effectively disintegrate WAS and can be combined with other physical techniques such as ultrasound for optimal results. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantitative analysis of chromosome condensation in fission yeast.

PubMed

Petrova, Boryana; Dehler, Sascha; Kruitwagen, Tom; Hériché, Jean-Karim; Miura, Kota; Haering, Christian H

2013-03-01

Chromosomes undergo extensive conformational rearrangements in preparation for their segregation during cell divisions. Insights into the molecular mechanisms behind this still poorly understood condensation process require the development of new approaches to quantitatively assess chromosome formation in vivo. In this study, we present a live-cell microscopy-based chromosome condensation assay in the fission yeast Schizosaccharomyces pombe. By automatically tracking the three-dimensional distance changes between fluorescently marked chromosome loci at high temporal and spatial resolution, we analyze chromosome condensation during mitosis and meiosis and deduct defined parameters to describe condensation dynamics. We demonstrate that this method can determine the contributions of condensin, topoisomerase II, and Aurora kinase to mitotic chromosome condensation. We furthermore show that the assay can identify proteins required for mitotic chromosome formation de novo by isolating mutants in condensin, DNA polymerase ε, and F-box DNA helicase I that are specifically defective in pro-/metaphase condensation. Thus, the chromosome condensation assay provides a direct and sensitive system for the discovery and characterization of components of the chromosome condensation machinery in a genetically tractable eukaryote.

Quantitative Analysis of Chromosome Condensation in Fission Yeast

PubMed Central

Petrova, Boryana; Dehler, Sascha; Kruitwagen, Tom; Hériché, Jean-Karim; Miura, Kota

2013-01-01

Chromosomes undergo extensive conformational rearrangements in preparation for their segregation during cell divisions. Insights into the molecular mechanisms behind this still poorly understood condensation process require the development of new approaches to quantitatively assess chromosome formation in vivo. In this study, we present a live-cell microscopy-based chromosome condensation assay in the fission yeast Schizosaccharomyces pombe. By automatically tracking the three-dimensional distance changes between fluorescently marked chromosome loci at high temporal and spatial resolution, we analyze chromosome condensation during mitosis and meiosis and deduct defined parameters to describe condensation dynamics. We demonstrate that this method can determine the contributions of condensin, topoisomerase II, and Aurora kinase to mitotic chromosome condensation. We furthermore show that the assay can identify proteins required for mitotic chromosome formation de novo by isolating mutants in condensin, DNA polymerase ε, and F-box DNA helicase I that are specifically defective in pro-/metaphase condensation. Thus, the chromosome condensation assay provides a direct and sensitive system for the discovery and characterization of components of the chromosome condensation machinery in a genetically tractable eukaryote. PMID:23263988

An in-vitro approach for water quality determination: activation of NF-κB as marker for cancer-related stress responses induced by anthropogenic pollutants of drinking water.

PubMed

Spitta, Luis F; Diegeler, Sebastian; Baumstark-Khan, Christa; Hellweg, Christine E

2018-02-01

Epidemiological studies show that there is a link between urban water pollution and increase in human morbidity and mortality. With the increase in number of new substances arising from the chemical, pharmaceutical, and agricultural industries, there is an urgent need to develop biological test systems for fast evaluation of potential risks to humans and the environmental ecosystems. Here, a combined cellular reporter assay based on the cellular survival and the stress-induced activation of the survival-promoting factor nuclear factor κB (NF-κB) and its use for the detection of cytotoxicity and cancer-related stress responses is presented. A total of 14 chemicals that may be found in trace-amounts in ground water levels are applied and tested with the presented assay. The project is embedded within the joint research project TOX-BOX which aims to develop a harmonized testing strategy for risk management of anthropogenic trace substances in potable water. The assay identified carbendazim as a NF-κB-activating agent in mammalian cells.

The use of spent mushroom compost to enhance the ability of Atriplex halimus to phytoremediate contaminated mine soils.

PubMed

Frutos, Iván; García-Delgado, Carlos; Cala, Victoria; Gárate, Agustin; Eymar, Enrique

2017-05-01

The mushroom cultivation industry produces a huge amount of spent mushroom compost (SMC), a wide world agricultural organic waste which causes serious environmental problems. However, this cheap organic waste could be useful in the remediation of contaminated soils. The aim of this work was to assess the potential of SMC in combination with the native shrub Atriplex halimus, to phytoremediate two mine soils contaminated with Cd, Pb and Cu. Firstly, to minimize metal availability in the soil, the optimal doses of SMC were determined. Secondly, a phytoremediation assay in greenhouse conditions was carried out to test the effects of A. halimus in combination with SMC at different doses. The results showed the ability of SMC to reduce soil acidity, the mobility of the metals and the enhancement of A. halimus growth. SMC promoted metal immobilization in the root of A. halimus and decreased the translocation from the roots to the shoots. The combination of SMC amendment and A. halimus produced phytostabilization of the metals in the mine soils assayed. In conclusion, SMC represents an adequate organic solid waste which in combination with A. halimus can reduce the adverse impact caused by the high mobility of metals in acid mine soils.

Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion

USGS Publications Warehouse

Haley, Nicholas J.; Siepker, Chris; Walter, W. David; Thomsen, Bruce V.; Greenlee, Justin J.; Lehmkuhl, Aaron D.; Richt, Jürgen a.

2016-01-01

Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since spread to cervids in 23 states, two Canadian provinces, and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of its spread; this is especially true in cases of relocation/reintroduction of farmed or free-ranging deer and elk or surveillance studies of private or protected herds, where depopulation is contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay by using recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brush samples collected antemortem from farmed white-tailed deer (n = 409). Antemortem findings were then compared to results from ante- and postmortem samples (RAMALT, brainstem, and medial retropharyngeal lymph nodes) evaluated by using the current gold standard in vitro assay, immunohistochemistry (IHC) analysis. We hypothesized that the sensitivity of RT-QuIC would be comparable to IHC analysis in antemortem tissues and would correlate with both the genotype and the stage of clinical disease. Our results showed that RAMALT testing by RT-QuIC assay had the highest sensitivity (69.8%) compared to that of postmortem testing, with a specificity of >93.9%. These data suggest that RT-QuIC, like IHC analysis, is an effective assay for detection of PrPCWD in rectal biopsy specimens and other antemortem samples and, with further research to identify more sensitive tissues, bodily fluids, or experimental conditions, has potential for large-scale and rapid automated testing for CWD diagnosis.

In-Situ Assays Using a New Advanced Mathematical Algorithm - 12400

DOE Office of Scientific and Technical Information (OSTI.GOV)

Oginni, B.M.; Bronson, F.L.; Field, M.B.

2012-07-01

Current mathematical efficiency modeling software for in-situ counting, such as the commercially available In-Situ Object Calibration Software (ISOCS), typically allows the description of measurement geometries via a list of well-defined templates which describe regular objects, such as boxes, cylinder, or spheres. While for many situations, these regular objects are sufficient to describe the measurement conditions, there are occasions in which a more detailed model is desired. We have developed a new all-purpose geometry template that can extend the flexibility of current ISOCS templates. This new template still utilizes the same advanced mathematical algorithms as current templates, but allows the extensionmore » to a multitude of shapes and objects that can be placed at any location and even combined. In addition, detectors can be placed anywhere and aimed at any location within the measurement scene. Several applications of this algorithm to in-situ waste assay measurements, as well as, validations of this template using Monte Carlo calculations and experimental measurements are studied. Presented in this paper is a new template of the mathematical algorithms for evaluating efficiencies. This new template combines all the advantages of the ISOCS and it allows the use of very complex geometries, it also allows stacking of geometries on one another in the same measurement scene and it allows the detector to be placed anywhere in the measurement scene and pointing in any direction. We have shown that the template compares well with the previous ISOCS software within the limit of convergence of the code, and also compare well with the MCNPX and measured data within the joint uncertainties for the code and the data. The new template agrees with ISOCS to within 1.5% at all energies. It agrees with the MCNPX to within 10% at all energies and it agrees with most geometries within 5%. It finally agrees with measured data to within 10%. This mathematical algorithm can now be used for quickly and accurately evaluating efficiencies for wider range of gamma-ray spectroscopy applications. (authors)« less

Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2.

PubMed

Yang, Yang; Qin, Xiaodong; Sun, Yingjun; Cong, Guozheng; Li, Yanmin; Zhang, Zhidong

2017-01-01

Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 10 2 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 10 2 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.

Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

PubMed Central

Yang, Yang; Qin, Xiaodong; Sun, Yingjun; Cong, Guozheng; Li, Yanmin

2017-01-01

Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2. PMID:28424790

Evaluation of a microarray-based genotyping assay for the rapid detection of cytochrome P450 2C19 *2 and *3 polymorphisms from whole blood using nanoparticle probes.

PubMed

Buchan, Blake W; Peterson, Jess F; Cogbill, Christopher H; Anderson, Dennis K; Ledford, Joellen S; White, Mary N; Quigley, Neil B; Jannetto, Paul J; Ledeboer, Nathan A

2011-10-01

Numerous drugs such as clopidogrel have been developed to reduce coagulation or inhibit platelet function. The hepatic cytochrome P450 (CYP) pathway is involved in the conversion of clopidogrel to its active metabolite. A recent black-box warning was included in the clopidogrel package insert indicating a significant clinical link between specific CYP2C19 genetic variants and poor metabolism of clopidogrel. Of these variants, *2 and *3 are the most common and are associated with complete loss of enzyme activity. In patients who are carriers of a CYP2C19 *2 or *3 allele, the conversion of clopidogrel to its active metabolite may be reduced, which can lead to ischemic events and negative consequence for the patient. We examined the ability of the Verigene CLO assay (Nanosphere, Northbrook, IL) to identify CYP2C19 *2 and *3 polymorphisms in 1,286 unique whole blood samples. The Verigene CLO assay accurately identified homozygous and heterozygous *2 and *3 phenotypes with a specificity of 100% and a final call rate of 99.7%. The assay is fully automated and can produce a result in approximately 3.5 hours.

Biomethanation potential of macroalgae Ulva spp. and Gracilaria spp. and in co-digestion with waste activated sludge.

PubMed

Costa, J C; Gonçalves, P R; Nobre, A; Alves, M M

2012-06-01

Biochemical methane potential of four species of Ulva and Gracilaria genus was assessed in batch assays at mesophilic temperature. The results indicate a higher specific methane production (per volatile solids) for one of the Ulva sp. compared with other macroalgae and for tests running with 2.5% of total solids (196±9 L CH(4) kg(-1)VS). Considering that macroalgae can potentially be a post treatment of municipal wastewater for nutrients removal, co-digestion of macroalgae with waste activated sludge (WAS) was assessed. The co-digestion of macroalgae (15%) with WAS (85%) is feasible at a rate of methane production 26% higher than WAS alone without decreasing the overall biodegradability of the substrate (42-45% methane yield). The use of anoxic marine sediment as inoculum had no positive effect on the methane production in batch assays. The limiting step of the overall anaerobic digestion process was the hydrolysis. Copyright © 2012 Elsevier Ltd. All rights reserved.

Metrological tests of a 200 L calibration source for HPGE detector systems for assay of radioactive waste drums.

PubMed

Boshkova, T; Mitev, K

2016-03-01

In this work we present test procedures, approval criteria and results from two metrological inspections of a certified large volume (152)Eu source (drum about 200L) intended for calibration of HPGe gamma assay systems used for activity measurement of radioactive waste drums. The aim of the inspections was to prove the stability of the calibration source during its working life. The large volume source was designed and produced in 2007. It consists of 448 identical sealed radioactive sources (modules) apportioned in 32 transparent plastic tubes which were placed in a wooden matrix which filled the drum. During the inspections the modules were subjected to tests for verification of their certified characteristics. The results show a perfect compliance with the NIST basic guidelines for the properties of a radioactive certified reference material (CRM) and demonstrate the stability of the large volume CRM-drum after 7 years of operation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Evaluating pharmaceutical waste disposal in pediatric units.

PubMed

Almeida, Maria Angélica Randoli de; Wilson, Ana Maria Miranda Martins; Peterlini, Maria Angélica Sorgini

2016-01-01

To verify the disposal of pharmaceutical waste performed in pediatric units. A descriptive and observational study conducted in a university hospital. The convenience sample consisted of pharmaceuticals discarded during the study period. Handling and disposal during preparation and administration were observed. Data collection took place at pre-established times and was performed using a pre-validated instrument. 356 drugs disposals were identified (35.1% in the clinic, 31.8% in the intensive care unit, 23.8% in the surgical unit and 9.3% in the infectious diseases unit). The most discarded pharmacological classes were: 22.7% antimicrobials, 14.8% electrolytes, 14.6% analgesics/pain killers, 9.5% diuretics and 6.7% antiulcer agents. The most used means for disposal were: sharps' disposable box with a yellow bag (30.8%), sink drain (28.9%), sharps' box with orange bag (14.3%), and infectious waste/bin with a white bag (10.1%). No disposal was identified after drug administration. A discussion of measures that can contribute to reducing (healthcare) waste volume with the intention of engaging reflective team performance and proper disposal is necessary. Verificar o descarte dos resíduos de medicamentos realizado em unidades pediátricas. Estudo descritivo e observacional, realizado em um hospital universitário. A amostra de conveniência foi constituída pelos medicamentos descartados durante o período de estudo. Observaram-se a manipulação e o descarte durante o preparo e a administração. A coleta dos dados ocorreu em horários preestabelecidos e realizada por meio de instrumento pré-validado. Identificaram-se 356 descartes de medicamentos (35,1% na clínica, 31,8% na unidade de cuidados intensivos, 23,8% na cirúrgica e 9,3% na infectologia). As classes farmacológicas mais descartadas foram: 22,7% antimicrobianos, 14,8% eletrólitos, 14,6% analgésicos, 9,5% diuréticos e 6,7% antiulcerosos. Vias mais utilizadas: caixa descartável para perfurocortante com saco amarelo (30,8%), ralo da pia (28,9%), caixa de perfurocortante com saco laranja (14,3%) e lixeira infectante com saco branco (10,1%). Não foi identificado descarte após a administração dos fármacos. Faz-se necessária a discussão de medidas que contribuam para a redução do volume de resíduos, com o intuito de engajar a atuação reflexiva da equipe e o descarte adequado.

Interlaboratory comparison program for nondestructive assay of prototype uranium reference materials

DOE Office of Scientific and Technical Information (OSTI.GOV)

Trahey, N.M.; Smith, M.M.; Voeks, A.M.

The US Department of Energy (DOE), New Brunswick Laboratory (NBS), designed and administered an interlaboratory comparison program based on the measurement of NBL-produced prototype uranium nondestructive assay (NDA) reference materials for scrap and waste. The objectives of the program were to evaluate the reliability of NDA techniques as applied to nuclear safeguards materials control and accountability needs and to investigate the feasibility of providing practical NDA scrap and waste reference materials for use throughout the nuclear safeguards community. Fourteen facilities representing seven DOE contractors, four US Nuclear Regulatory Commission (NRC) licensees, one EURATOM Laboratory, and NBL, participated in this program.more » Three stable, well-characterized uranium reference materials were developed and certified for this program. Synthetic calcined ash, cellulose fiber, and ion-exchange resin simulate selected uranium scrap and waste forms which are often encountered in fabrication and recovery operations. The synthetic calcined ash represents an intermediate density inorganic matrix while the cellulose fiber and ion-exchange resin are representative of low-density organic matrices. The materials, containing from 0 to 13% uranium enriched at 93% /sup 235/U, were sealed in specially selected containers. Nineteen prototype reference samples, plus three empty containers, one to accompany each set, was circulated to the participants between August 1979 and May 1984. Triplicate measurements for /sup 235/U on each of the 19 filled containers were required. In addition, participants could opt to perform modular configuration measurements using containers from Sets IIA and IIB to simulate non-homogeneously dispersed uranium in waste containers. All data were reported to NBL for evaluation.« less

A global health partnership's use of time-limited support to catalyze health practice change: the case of GAVI's Injection Safety Support.

PubMed

Levin, Ann; Fang, Arnold; Hansen, Peter M; Pyle, David; Dia, Ousmane; Schwalbe, Nina

2010-09-27

This paper presents the findings of a study to assess the effectiveness and sustainability of a GAVI (Global Alliance of Vaccines and Immunization) sponsored, time-limited Injection Safety (INS) support. The support came in two forms: 1) in-kind, in the form of AD syringes and safety boxes, and 2) in cash, for those countries that already had a secure, multi-year source of AD syringes and safety boxes, but proposed to use INS support to strengthen their injection safety activities. In total, GAVI gave INS support for a three-year period to 58 countries: 46 with commodities and 12 with cash support. To identify variables that might be associated with financial sustainability, frequencies and cross-tabulations were run against various programmatic and socio-economic variables in the 58 countries. All but two of the 46 commodity-recipient countries were able to replace and sustain the use of AD syringes and safety boxes after the end of their GAVI INS support despite the fact that standard disposable syringes are less costly than ADs (10-15 percent differential). In addition, all 12 cash-recipient countries continued to use AD syringes and safety boxes in their immunization programs in the years following GAVI INS assistance. At the same time, countries were often not prepared for the increased waste management requirements associated with the use of the syringes, suggesting the importance of anticipating challenges with the introduction of new technologies. The sustained use of AD syringes in countries receiving injection safety support from GAVI, in a majority of cases through government financing, following the completion of three years of time-limited support, represents an early indication of how GHPs can contribute to improved health outcomes in immunization safety in the world's poorest countries in a sustainable way.

Cold shock protein YB-1 is involved in hypoxia-dependent gene transcription

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rauen, Thomas; Frye, Bjoern C.; Pneumology, University Medical Center, University of Freiburg, Freiburg

Hypoxia-dependent gene regulation is largely orchestrated by hypoxia-inducible factors (HIFs), which associate with defined nucleotide sequences of hypoxia-responsive elements (HREs). Comparison of the regulatory HRE within the 3′ enhancer of the human erythropoietin (EPO) gene with known binding motifs for cold shock protein Y-box (YB) protein-1 yielded strong similarities within the Y-box element and 3′ adjacent sequences. DNA binding assays confirmed YB-1 binding to both, single- and double-stranded HRE templates. Under hypoxia, we observed nuclear shuttling of YB-1 and co-immunoprecipitation assays demonstrated that YB-1 and HIF-1α physically interact with each other. Cellular YB-1 depletion using siRNA significantly induced hypoxia-dependent EPOmore » production at both, promoter and mRNA level. Vice versa, overexpressed YB-1 significantly reduced EPO-HRE-dependent gene transcription, whereas this effect was minor under normoxia. HIF-1α overexpression induced hypoxia-dependent gene transcription through the same element and accordingly, co-expression with YB-1 reduced HIF-1α-mediated EPO induction under hypoxic conditions. Taken together, we identified YB-1 as a novel binding factor for HREs that participates in fine-tuning of the hypoxia transcriptome. - Highlights: • Hypoxia drives nuclear translocation of cold shock protein YB-1. • YB-1 physically interacts with hypoxia-inducible factor (HIF)-1α. • YB-1 binds to the hypoxia-responsive element (HRE) within the erythropoietin (EPO) 3′ enhancer. • YB-1 trans-regulates transcription of hypoxia-dependent genes such as EPO and VEGF.« less

Valproic acid attenuates skeletal muscle wasting by inhibiting C/EBPβ-regulated atrogin1 expression in cancer cachexia.

PubMed

Sun, Rulin; Zhang, Santao; Hu, Wenjun; Lu, Xing; Lou, Ning; Yang, Zhende; Chen, Shaoyong; Zhang, Xiaoping; Yang, Hongmei

2016-07-01

Muscle wasting is the hallmark of cancer cachexia and is associated with poor quality of life and increased mortality. Valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, has important biological effects in the treatment of muscular dystrophy. To verify whether VPA could ameliorate muscle wasting induced by cancer cachexia, we explored the role of VPA in two cancer cachectic mouse models [induced by colon-26 (C26) adenocarcinoma or Lewis lung carcinoma (LLC)] and atrophied C2C12 myotubes [induced by C26 cell conditioned medium (CCM) or LLC cell conditioned medium (LCM)]. Our data demonstrated that treatment with VPA increased the mass and cross-sectional area of skeletal muscles in tumor-bearing mice. Furthermore, treatment with VPA also increased the diameter of myotubes cultured in conditioned medium. The skeletal muscles in cachectic mice or atrophied myotubes treated with VPA exhibited reduced levels of CCAAT/enhancer binding protein beta (C/EBPβ), resulting in atrogin1 downregulation and the eventual alleviation of muscle wasting and myotube atrophy. Moreover, atrogin1 promoter activity in myotubes was stimulated by CCM via activating the C/EBPβ-responsive cis-element and subsequently inhibited by VPA. In contrast to the effect of VPA on the levels of C/EBPβ, the levels of inactivating forkhead box O3 (FoxO3a) were unaffected. In summary, VPA attenuated muscle wasting and myotube atrophy and reduced C/EBPβ binding to atrogin1 promoter locus in the myotubes. Our discoveries indicate that HDAC inhibition by VPA might be a promising new approach for the preservation of skeletal muscle in cancer cachexia. Copyright © 2016 the American Physiological Society.

Chemical and Biological Investigation of Olive Mill Waste Water - OMWW Secoiridoid Lactones.

PubMed

Vougogiannopoulou, Konstantina; Angelopoulou, Maria T; Pratsinis, Harris; Grougnet, Raphaël; Halabalaki, Maria; Kletsas, Dimitris; Deguin, Brigitte; Skaltsounis, Leandros A

2015-08-01

Olive mill waste water is the major byproduct of the olive oil industry containing a range of compounds related to Olea europaea and olive oil constituents. Olive mill waste water comprises an important environmental problem in olive oil producing countries, but it is also a valuable material for the isolation of high added value compounds. In this study, an attempt to investigate the secoiridoid content of olive mill waste water is described with the aid of ultrahigh-performance liquid chromatography-electrospray ionization (±)-high-resolution mass spectrometry and centrifugal partition chromatography methods. In total, seven secoiridoid lactones were isolated, four of which are new natural products. This is the first time that a conjugate of hydroxytyrosol and a secoiridoid lactone has been isolated from olive mill waste water and structurally characterized. Furthermore, the range of isolated compounds allowed for the proposal of a hypothesis for the biotransformation of olive secoiridoids during the production of olive mill waste water. Finally, the ability of the representative compounds to reduce the intracellular reactive oxygen species was assessed with the dichlorofluorescein assay in conjunction with the known antioxidant agent hydroxytyrosol. Georg Thieme Verlag KG Stuttgart · New York.

Performance of NDA techniques on a vitrified waste form

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hurd, J.R.; Veazey, G.W.; Prettyman, T.H.

1997-11-01

Rocky Flats Environmental Technology Site (RFETS) is currently considering the use of vitrified transuranic (TRU)-waste forms for the final disposition of several waste materials. To date, however, little nondestructive assay (NDA) data have been acquired in the general NDA community to assist in this endeavor. This paper describes the efforts to determine constraints and operating parameters for using NDA instrumentation on vitrified waste. The present study was conducted on a sample composed of a plutonium-contaminated ash, similar to that found in the RFETS inventory, and a borosilicate-based glass. The vitrified waste item was fabricated at Los Alamos National Laboratory (LANL)more » using methods and equipment similar to those being proposed by RFETS to treat their ash material. The focus of this study centered on the segmented gamma scanner (SGS) with 1/2-inch collimation, a technique that is presently available at RFETS. The accuracy and precision of SGS technology was evaluated, with particular attention to bias issues involving matrix geometry, homogeneity, and attenuation. Tomographic gamma scanning was utilized in the determination of the waste form homogeneity. A thermal neutron technique was also investigated and comparisons made with the gamma results.« less

Validated electrochemical and chromatographic quantifications of some antibiotic residues in pharmaceutical industrial waste water.

PubMed

Ibrahim, Heba K; Abdel-Moety, Mona M; Abdel-Gawad, Sherif A; Al-Ghobashy, Medhat A; Kawy, Mohamed Abdel

2017-03-01

Realistic implementation of ion selective electrodes (ISEs) into environmental monitoring programs has always been a challenging task. This could be largely attributed to difficulties in validation of ISE assay results. In this study, the electrochemical response of amoxicillin trihydrate (AMX), ciprofloxacin hydrochloride (CPLX), trimethoprim (TMP), and norfloxacin (NFLX) was studied by the fabrication of sensitive membrane electrodes belonging to two types of ISEs, which are polyvinyl chloride (PVC) membrane electrodes and glassy carbon (GC) electrodes. Linear response for the membrane electrodes was in the concentration range of 10 -5 -10 -2  mol/L. For the PVC membrane electrodes, Nernstian slopes of 55.1, 56.5, 56.5, and 54.0 mV/decade were achieved over a pH 4-8 for AMX, CPLX, and NFLX, respectively, and pH 3-6 for TMP. On the other hand, for GC electrodes, Nernstian slopes of 59.1, 58.2, 57.0, and 58.2 mV/decade were achieved over pH 4-8 for AMX, CPLX, and NFLX, respectively, and pH 3-6 for TMP. In addition to assay validation to international industry standards, the fabricated electrodes were also cross-validated relative to conventional separation techniques; high performance liquid chromatography (HPLC), and thin layer chromatography (TLC)-densitometry. The HPLC assay was applied in concentration range of 0.5-10.0 μg/mL, for all target analytes. The TLC-densitometry was adopted over a concentration range of 0.3-1.0 μg/band, for AMX, and 0.1-0.9 μg/band, for CPLX, NFLX, and TMP. The proposed techniques were successfully applied for quantification of the selected drugs either in pure form or waste water samples obtained from pharmaceutical plants. The actual waste water samples were subjected to solid phase extraction (SPE) for pretreatment prior to the application of chromatographic techniques (HPLC and TLC-densitometry). On the other hand, the fabricated electrodes were successfully applied for quantification of the antibiotic residues in actual waste water samples without any pretreatment. This finding assures the suitability of the fabricated ISEs for environmental analysis.

Two High Throughput Screen Assays for Measurement of TNF-α in THP-1 Cells

PubMed Central

Leister, Kristin P; Huang, Ruili; Goodwin, Bonnie L; Chen, Andrew; Austin, Christopher P; Xia, Menghang

2011-01-01

Tumor Necrosis Factor-α (TNF-α), a secreted cytokine, plays an important role in inflammatory diseases and immune disorders, and is a potential target for drug development. The traditional assays for detecting TNF-α, enzyme linked immunosorbent assay (ELISA) and radioimmunoassay, are not suitable for the large size compound screens. Both assays suffer from a complicated protocol, multiple plate wash steps and/or excessive radioactive waste. A simple and quick measurement of TNF-α production in a cell based assay is needed for high throughput screening to identify the lead compounds from the compound library. We have developed and optimized two homogeneous TNF-α assays using the HTRF (homogeneous time resolved fluorescence) and AlphaLISA assay formats. We have validated the HTRF based TNF-α assay in a 1536-well plate format by screening a library of 1280 pharmacologically active compounds. The active compounds identified from the screen were confirmed in the AlphaLISA TNF-α assay using a bead-based technology. These compounds were also confirmed in a traditional ELISA assay. From this study, several beta adrenergic agonists have been identified as TNF-α inhibitors. We also identified several novel inhibitors of TNF-α, such as BTO-1, CCG-2046, ellipticine, and PD 169316. The results demonstrated that both homogeneous TNF-α assays are robust and suitable for high throughput screening. PMID:21643507

Characterization of TM8, a MADS-box gene expressed in tomato flowers.

PubMed

Daminato, Margherita; Masiero, Simona; Resentini, Francesca; Lovisetto, Alessandro; Casadoro, Giorgio

2014-11-30

The identity of flower organs is specified by various MIKC MADS-box transcription factors which act in a combinatorial manner. TM8 is a MADS-box gene that was isolated from the floral meristem of a tomato mutant more than twenty years ago, but is still poorly known from a functional point of view in spite of being present in both Angiosperms and Gymnosperms, with some species harbouring more than one copy of the gene. This study reports a characterization of TM8 that was carried out in transgenic tomato plants with altered expression of the gene. Tomato plants over-expressing either TM8 or a chimeric repressor form of the gene (TM8:SRDX) were prepared. In the TM8 up-regulated plants it was possible to observe anomalous stamens with poorly viable pollen and altered expression of several floral identity genes, among them B-, C- and E-function ones, while no apparent morphological modifications were visible in the other whorls. Oblong ovaries and fruits, that were also parthenocarpic, were obtained in the plants expressing the TM8:SRDX repressor gene. Such ovaries showed modified expression of various carpel-related genes. No apparent modifications could be seen in the other flower whorls. The latter plants had also epinastic leaves and malformed flower abscission zones. By using yeast two hybrid assays it was possible to show that TM8 was able to interact in yeast with MACROCALIX. The impact of the ectopically altered TM8 expression on the reproductive structures suggests that this gene plays some role in the development of the tomato flower. MACROCALYX, a putative A-function MADS-box gene, was expressed in all the four whorls of fully developed flowers, and showed quantitative variations that were opposite to those of TM8 in the anomalous stamens and ovaries. Since the TM8 protein interacted in vitro only with the A-function MADS-box protein MACROCALYX, it seems that for the correct differentiation of the tomato reproductive structures possible interactions between TM8 and MACROCALYX proteins might be important.

Treatment of supermarket vegetable wastes to be used as alternative substrates in bioprocesses.

PubMed

Díaz, Ana Isabel; Laca, Amanda; Laca, Adriana; Díaz, Mario

2017-09-01

Fruits and vegetables have the highest wastage rates at retail and consumer levels. These wastes have promising potential for being used as substrates in bioprocesses. However, an effective hydrolysis of carbohydrates that form these residues has to be developed before the biotransformation. In this work, vegetable wastes from supermarket (tomatoes, green peppers and potatoes) have been separately treated by acid, thermal and enzymatic hydrolysis processes in order to maximise the concentration of fermentable sugars in the final broth. For all substrates, thermal and enzymatic processes have shown to be the most effective. A new combined hydrolysis procedure including these both treatments was also assayed and the enzymatic step was successfully modelled. With this combined hydrolysis, the percentage of reducing sugars extracted was increased, in comparison with the amount extracted from non-hydrolysed samples, approximately by 30% in the case of tomato and green peeper wastes. For potato wastes this percentage increased from values lower than 1% to 77%. In addition, very low values of fermentation inhibitors were found in the final broth. Copyright © 2017. Published by Elsevier Ltd.

Effects of inoculum source and co-digestion strategies on anaerobic digestion of residues generated in the treatment of waste vegetable oils.

PubMed

Hidalgo, Dolores; Martín-Marroquín, Jesús M

2014-09-01

This work aims at selecting a suitable strategy to improve the performance of the anaerobic digestion of residues generated in the treatment of waste vegetable oils (WVO). Biochemical methane potential (BMP) assays were conducted at 35 °C to evaluate the effects of substrate mix ratio between a mixture of WVO residues (M) and pig manure (PM) co-digesting by using different inocula. Inoculum from an industrial digester fed with organic waste from hotels, restaurants and catering leftovers (HORECA) showed higher methanogenic activity (55.5 mLCH4 gVS(-1) d(-1)) than municipal wastewater treatment plant (mWWTP) inoculum (42.6 mL CH4 gVS(-1) d(-1)). Furthermore, the results showed that the resistance to WVO residues toxicity was higher for the HORECA sludge than for the mWWTP sludge. HORECA inoculum produced more biogas in all the assays. Moreover, the resulting biogas was of better quality, containing an average of 71.1% (SD = 1.6) methane compared to an average of 69.5% (SD = 1.2) methane for test with mWWTP sludge. The maximum degradation rate occurred at the higher PM mix ratio (M/PM:1/3), reaching 26.7 ± 4.3 mLCH4 gVS(-1) d(-1) for mWWTP inoculum, versus 42.0 ± 1,5 mLCH4 gVS(-1) d(-1) achieved for HORECA inoculum. A high reduction of volatile solids (between 70% and 81%) was obtained with both inocula at all M/PM ratios assayed (1/0, 1/3, 1/1 and 3/1 v/v) but, bearing in mind the operation of a full-scale anaerobic plant, the optimal scenario assayed corresponds to the ratio M/PM: 1/3 v/v where shorter lag periods will make it possible to operate at lower hydraulic retention times. Copyright © 2014 Elsevier Ltd. All rights reserved.

Optimization of extraction parameters on the antioxidant properties of banana waste.

PubMed

Toh, Pui Yee; Leong, Fei Shan; Chang, Sui Kiat; Khoo, Hock Eng; Yim, Hip Seng

2016-01-01

Banana is grown worldwide and consumed as ripe fruit or used for culinary purposes. Peels form about 18-33% of the whole fruit and are discarded as a waste product. With a view to exploiting banana peel as a source of valuable compounds, this study was undertaken to evaluate the effect of different extraction parameters on the antioxidant activities of the industrial by-product of banana waste (peel). Influence of different extraction parameters such as types of solvent, percentages of solvent, and extraction times on total phenolic content (TPC) and antioxidant activity of mature and green peels of Pisang Abu (PA), Pisang Berangan (PB), and Pisang Mas (PM) were investigated. The best extraction parameters were initially selected based on different percentages of ethanol (0-100% v/v), extraction time (1-5 hr), and extraction temperature (25-60°C) for extraction of antioxidants in the banana peels. Total phenolic content (TPC) was evaluated using Folin-Ciocalteu reagent assay while antioxidant activities (AA) of banana peel were accessed by DPPH, ABTS, and β-carotene bleaching (BCB) assays at optimum extraction conditions. Based on different extraction solvents and percentages of solvents used, 70% and 90% of acetone had yielded the highest TPC for the mature and green PA peels, respectively; 90% of ethanol and methanol has yielded the highest TPC for the mature and green PB peels, respectively; while 90% ethanol for the mature and green PM peels. Similar extraction conditions were found for the antioxidant activities for the banana peel assessed using DPPH assay except for green PB peel, which 70% methanol had contributed to the highest AA. Highest TPC and AA were obtained by applying 4, 1, and 2 hrs extraction for the peels of PA, PB and PM, respectively. The best extraction conditions were also used for determination of AAs using ABTS and β-carotene bleaching assays. Therefore, the best extraction conditions used have given the highest TPC and AAs. By-products of banana (peel) can be considered as a potential source of antioxidants in food and pharmaceutical industry.

Application of Response Surface Methodology for Optimization of Extracellular Glucoamylase Production by Candida guilliermondii.

PubMed

Mohamed, Lagzouli; Kettani, Youssfi El; Ali, Aitounejjar; Mohamed, Elyachioui; Mohamed, Jadal

2017-01-01

Glucoamylase is among the most important enzymes in biotechnology. The present study aims to determine better conditions for growth and glucoamylase production by Candida guilliermondii and to reduce the overall cost of the medium using Box-Behnken design with one central point and response surface methodology. Box-Behnken factorial design based on three levels was carried out to obtain optimal medium combination of five independent variables such as initial pH, soluble starch, CH4N2O, yeast extract and MgSO4. Forty one randomized mediums were incubated in flask on a rotary shaker at 105 rpm for 72 h at 30°C. The production of biomass was found to be pH and starch dependent, maximum production when the starch concentration was 8 g L-1 and the initial pH was 6, while maximum glucoamylase production was found at 6.5 of initial pH, 4 g L-1 yeast extract and 6 g L-1 starch, whereas yeast extract and urea were highly significant, but interacted negatively. Box-Behnken factorial design used for the analysis of treatment combinations gave a second-order polynomial regression model with R2 = 0.976 for Biomass and R2 = 0.981 for glucoamylase. The final biomass and glucoamylase activity obtained was very close to the calculated parameters according to the p-values (p<0.001), the predicted optimal parameters were confirmed and provides a basis for further studies in baking additives and in the valuation of starch waste products.

Fusion of microlitre water-in-oil droplets for simple, fast and green chemical assays.

PubMed

Chiu, S-H; Urban, P L

2015-08-07

A simple format for microscale chemical assays is proposed. It does not require the use of test tubes, microchips or microtiter plates. Microlitre-range (ca. 0.7-5.0 μL) aqueous droplets are generated by a commercial micropipette in a non-polar matrix inside a Petri dish. When two droplets are pipetted nearby, they spontaneously coalesce within seconds, priming a chemical reaction. Detection of the reaction product is accomplished by colorimetry, spectrophotometry, or fluorimetry using simple light-emitting diode (LED) arrays as the sources of monochromatic light, while chemiluminescence detection of the analytes present in single droplets is conducted in the dark. A smartphone camera is used as the detector. The limits of detection obtained for the developed in-droplet assays are estimated to be: 1.4 nmol (potassium permanganate by colorimetry), 1.4 pmol (fluorescein by fluorimetry), and 580 fmol (sodium hypochlorite by chemiluminescence detection). The format has successfully been used to monitor the progress of chemical and biochemical reactions over time with sub-second resolution. A semi-quantitative analysis of ascorbic acid using Tillman's reagent is presented. A few tens of individual droplets can be scanned in parallel. Rapid switching of the LED light sources with different wavelengths enables a spectral analysis of multiple droplets. Very little solid waste is produced. The assay matrix is readily recycled, thus the volume of liquid waste produced each time is also very small (typically, 1-10 μL per analysis). Various water-immiscible translucent liquids can be used as the reaction matrix: including silicone oil, 1-octanol as well as soybean cooking oil.

Development and characterization of an ex-vivo brain slice culture model of chronic wasting disease

USDA-ARS?s Scientific Manuscript database

Prion diseases have long incubation times in vivo, therefore, modeling the diseases ex-vivo will advance the development of rationale-based therapeutic strategies. An organotypic slice culture assay (POSCA) was recently developed for scrapie prions by inoculating mouse cerebellar brain slices with R...

In vitro assays for assessment of androgenic and estrogenic activity of defined mixtures and complex environment samples

EPA Science Inventory

Point sources of potentially endocrine active compounds to aquatic environments such as waste water treatment plants, pulp and paper mills, and animal feeding operations invariably contain complex mixtures of chemicals. The current study investigates the use of targeted in vitro ...

In vitro assays for assessment of androgenic and estrogenic activity in defined mixtures and complex environmental samples

EPA Science Inventory

Eflluents from sources such as waste water treatment plants and animal feeding operations invariably contain complex mixtures of chemicals. Recent research on effluent from cattle feeding operations in the US have linked morphological alterations in fish with in vitro androgenic ...

The Drosophila Juvenile Hormone Receptor Candidates Methoprene-tolerant (MET) and Germ Cell-expressed (GCE) Utilize a Conserved LIXXL Motif to Bind the FTZ-F1 Nuclear Receptor*

PubMed Central

Bernardo, Travis J.; Dubrovsky, Edward B.

2012-01-01

Juvenile hormone (JH) has been implicated in many developmental processes in holometabolous insects, but its mechanism of signaling remains controversial. We previously found that in Drosophila Schneider 2 cells, the nuclear receptor FTZ-F1 is required for activation of the E75A gene by JH. Here, we utilized insect two-hybrid assays to show that FTZ-F1 interacts with two JH receptor candidates, the bHLH-PAS paralogs MET and GCE, in a JH-dependent manner. These interactions are severely reduced when helix 12 of the FTZ-F1 activation function 2 (AF2) is removed, implicating AF2 as an interacting site. Through homology modeling, we found that MET and GCE possess a C-terminal α-helix featuring a conserved motif LIXXL that represents a novel nuclear receptor (NR) box. Docking simulations supported by two-hybrid experiments revealed that FTZ-F1·MET and FTZ-F1·GCE heterodimer formation involves a typical NR box-AF2 interaction but does not require the canonical charge clamp residues of FTZ-F1 and relies primarily on hydrophobic contacts, including a unique interaction with helix 4. Moreover, we identified paralog-specific features, including a secondary interaction site found only in MET. Our findings suggest that a novel NR box enables MET and GCE to interact JH-dependently with the AF2 of FTZ-F1. PMID:22249180

The newt (Cynops pyrrhogaster) RPE65 promoter: molecular cloning, characterization and functional analysis.

PubMed

Casco-Robles, Martin Miguel; Miura, Tomoya; Chiba, Chikafumi

2015-06-01

The adult newt has the ability to regenerate the neural retina following injury, a process achieved primarily by the retinal pigment epithelium (RPE). To deliver exogenous genes to the RPE for genetic manipulation of regenerative events, we isolated the newt RPE65 promoter region by genome walking. First, we cloned the 2.8 kb RPE65 promoter from the newt, Cynops pyrrhogaster. Sequence analysis revealed several conserved regulatory elements described previously in mouse and human RPE65 promoters. Second, having previously established an I-SceI-mediated transgenic protocol for the newt, we used it here to examine the -657 bp proximal promoter of RPE65. The promoter assay used with F0 transgenic newts confirmed transgene expression of mCherry fluorescent protein in the RPE. Using bioinformatic tools and the TRANSFAC database, we identified a 340 bp CpG island located between -635 and -296 bp in the promoter; this region contains response elements for the microphthalmia-associated transcription factor known as MITF (CACGTG, CATGTG), and E-boxes (CANNTG). Sex-determining region box 9 (or SOX9) response element previously reported in the regulation of RPE genes (including RPE65) was also identified in the newt RPE65 promoter. Third, we identified DNA motif boxes in the newt RPE65 promoter that are conserved among other vertebrates. The newt RPE65 promoter is an invaluable tool for site-specific delivery of exogenous genes or genetic manipulation systems for the study of retinal regeneration in this animal.

CHARACTERIZING THE 25-HYDROXYVITAMIN D STATUS OF TWO POPULATIONS OF FREE-RANGING EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA).

PubMed

Watson, Megan K; Byrd, John; Phillips, Christopher A; Allender, Matthew C

2017-09-01

Ultraviolet B radiation is recommended for captive reptiles to stimulate production of adequate levels of vitamin D; however, little is known regarding the vitamin D status in many free-ranging populations. Current reference ranges for vitamin D in eastern box turtles have not yet been established. Sixty free-ranging eastern box turtles (Terrapene carolina carolina) from two well-studied populations in Illinois (n = 24) and Tennessee (n = 36) were assayed for plasma vitamin D concentration in 2014. There were no significant differences in concentrations between individuals in Illinois (mean: 117.5 nM/L) and Tennessee (mean: 98.7 nM/L) (P = 0.129) populations. Similarly, there were no differences in concentrations based on age class (P = 0.533) or sex (P = 0.532). There was a significant correlation between UV at the time of capture and vitamin D concentrations (R = 0.301, P = 0.030). Vitamin D was not correlated with total calcium (R = 0.018, P = 0.89) or Ca : P ratio (R = 0.025, P = 0.85). Diseases in captive individuals, including secondary nutritional hyperparathyroidism, may commonly be associated with vitamin D deficiencies, and clinical intervention relies on reference data. Vitamin D supplementation may be recommended if animals are deemed to be deficient. Data obtained can be used to improve the care of captive and free-ranging turtles by providing reference ranges, as well as better characterize the health of wild populations.

A simple method for optimising transformation of non-parametric data: an illustration by reference to cortisol assays.

PubMed

Clark, James E; Osborne, Jason W; Gallagher, Peter; Watson, Stuart

2016-07-01

Neuroendocrine data are typically positively skewed and rarely conform to the expectations of a Gaussian distribution. This can be a problem when attempting to analyse results within the framework of the general linear model, which relies on assumptions that residuals in the data are normally distributed. One frequently used method for handling violations of this assumption is to transform variables to bring residuals into closer alignment with assumptions (as residuals are not directly manipulated). This is often attempted through ad hoc traditional transformations such as square root, log and inverse. However, Box and Cox (Box & Cox, ) observed that these are all special cases of power transformations and proposed a more flexible method of transformation for researchers to optimise alignment with assumptions. The goal of this paper is to demonstrate the benefits of the infinitely flexible Box-Cox transformation on neuroendocrine data using syntax in spss. When applied to positively skewed data typical of neuroendocrine data, the majority (~2/3) of cases were brought into strict alignment with Gaussian distribution (i.e. a non-significant Shapiro-Wilks test). Those unable to meet this challenge showed substantial improvement in distributional properties. The biggest challenge was distributions with a high ratio of kurtosis to skewness. We discuss how these cases might be handled, and we highlight some of the broader issues associated with transformation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

The MADS-box transcription factor FgMcm1 regulates cell identity and fungal development in Fusarium graminearum.

PubMed

Yang, Cui; Liu, Huiquan; Li, Guotian; Liu, Meigang; Yun, Yingzi; Wang, Chenfang; Ma, Zhonghua; Xu, Jin-Rong

2015-08-01

In eukaryotic cells, MADS-box genes are known to play major regulatory roles in various biological processes by combinatorial interactions with other transcription factors. In this study, we functionally characterized the FgMCM1 MADS-box gene in Fusarium graminearum, the causal agent of wheat and barley head blight. Deletion of FgMCM1 resulted in the loss of perithecium production and phialide formation. The Fgmcm1 mutant was significantly reduced in virulence, deoxynivalenol biosynthesis and conidiation. In yeast two-hybrid assays, FgMcm1 interacted with Mat1-1-1 and Fst12, two transcription factors important for sexual reproduction. Whereas Fgmcm1 mutants were unstable and produced stunted subcultures, Fgmcm1 mat1-1-1 but not Fgmcm1 fst12 double mutants were stable. Furthermore, spontaneous suppressor mutations occurred frequently in stunted subcultures to recover growth rate. Ribonucleic acid sequencing analysis indicated that a number of sexual reproduction-related genes were upregulated in stunted subcultures compared with the Fgmcm1 mutant, which was downregulated in the expression of genes involved in pathogenesis, secondary metabolism and conidiation. We also showed that culture instability was not observed in the Fvmcm1 mutants of the heterothallic Fusarium verticillioides. Overall, our data indicate that FgMcm1 plays a critical role in the regulation of cell identity, sexual and asexual reproduction, secondary metabolism and pathogenesis in F. graminearum. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

The snoRNA domain of vertebrate telomerase RNA functions to localize the RNA within the nucleus.

PubMed Central

Lukowiak, A A; Narayanan, A; Li, Z H; Terns, R M; Terns, M P

2001-01-01

Telomerase RNA is an essential component of the ribonucleoprotein enzyme involved in telomere length maintenance, a process implicated in cellular senescence and cancer. Vertebrate telomerase RNAs contain a box H/ACA snoRNA motif that is not required for telomerase activity in vitro but is essential in vivo. Using the Xenopus oocyte system, we have found that the box H/ACA motif functions in the subcellular localization of telomerase RNA. We have characterized the transport and biogenesis of telomerase RNA by injecting labeled wild-type and variant RNAs into Xenopus oocytes and assaying nucleocytoplasmic distribution, intranuclear localization, modification, and protein binding. Although yeast telomerase RNA shares characteristics of spliceosomal snRNAs, we show that human telomerase RNA is not associated with Sm proteins or efficiently imported into the nucleus. In contrast, the transport properties of vertebrate telomerase RNA resemble those of snoRNAs; telomerase RNA is retained in the nucleus and targeted to nucleoli. Furthermore, both nuclear retention and nucleolar localization depend on the box H/ACA motif. Our findings suggest that the H/ACA motif confers functional localization of vertebrate telomerase RNAs to the nucleus, the compartment where telomeres are synthesized. We have also found that telomerase RNA localizes to Cajal bodies, intranuclear structures where it is thought that assembly of various cellular RNPs takes place. Our results identify the Cajal body as a potential site of telomerase RNP biogenesis. PMID:11780638

Functional importance of evolutionally conserved Tbx6 binding sites in the presomitic mesoderm-specific enhancer of Mesp2.

PubMed

Yasuhiko, Yukuto; Kitajima, Satoshi; Takahashi, Yu; Oginuma, Masayuki; Kagiwada, Harumi; Kanno, Jun; Saga, Yumiko

2008-11-01

The T-box transcription factor Tbx6 controls the expression of Mesp2, which encodes a basic helix-loop-helix transcription factor that has crucial roles in somitogenesis. In cultured cells, Tbx6 binding to the Mesp2 enhancer region is essential for the activation of Mesp2 by Notch signaling. However, it is not known whether this binding is required in vivo. Here we report that an Mesp2 enhancer knockout mouse bearing mutations in two crucial Tbx6 binding sites does not express Mesp2 in the presomitic mesoderm. This absence leads to impaired skeletal segmentation identical to that reported for Mesp2-null mice, indicating that these Tbx6 binding sites are indispensable for Mesp2 expression. T-box binding to the consensus sequences in the Mesp2 upstream region was confirmed by chromatin immunoprecipitation assays. Further enhancer analyses indicated that the number and spatial organization of the T-box binding sites are critical for initiating Mesp2 transcription via Notch signaling. We also generated a knock-in mouse in which the endogenous Mesp2 enhancer was replaced by the core enhancer of medaka mespb, an ortholog of mouse Mesp2. The homozygous enhancer knock-in mouse was viable and showed normal skeletal segmentation, indicating that the medaka mespb enhancer functionally replaced the mouse Mesp2 enhancer. These results demonstrate that there is significant evolutionary conservation of Mesp regulatory mechanisms between fish and mice.

The CCAAT box in the proximal SERCA2 gene promoter regulates basal and stress-induced transcription in cardiomyocytes.

PubMed

Fragoso-Medina, Jorge; Rodriguez, Gabriela; Zarain-Herzberg, Angel

2018-05-01

The cardiac sarco/endoplasmic reticulum Ca 2+ -ATPase-2a (SERCA2a) is vital for the correct handling of calcium concentration in cardiomyocytes. Recent studies showed that the induction of endoplasmic reticulum (ER) stress (ERS) with the SERCA2 inhibitor Thapsigargin (Tg) increases the mRNA and protein levels of SERCA2a. The SERCA2 gene promoter contains an ERS response element (ERSE) at position -78 bp that is conserved among species and might transcriptionally regulate SERCA2 gene expression. However, its involvement in SERCA2 basal and calcium-mediated transcriptional activation has not been elucidated. In this work, we show that in cellular cultures of neonatal rat ventricular myocytes, the treatment with Tg or the calcium ionophore A23187 increases the SERCA2a mRNA and protein abundance, as well as the transcriptional activity of two chimeric human SERCA2 gene constructs, containing -254 and -2579 bp of 5'-regulatory region cloned in the pGL3-basic vector and transiently transfected in cultured cardiomyocytes. We found that the ERSE present in the SERCA2 proximal promoter contains a CCAAT box that is involved in basal and ERS-mediated hSERCA2 transcriptional activation. The EMSA results showed that the CCAAT box present in the ERSE recruits the NF-Y transcription factor. Additionally, by ChIP assays, we confirmed in vivo binding of NF-Y and C/EBPβ transcription factors to the SERCA2 gene proximal promoter.

The Wnt-1 (int-1) oncogene promoter and its mechanism of activation by insertion of proviral DNA of the mouse mammary tumor virus.

PubMed Central

Nusse, R; Theunissen, H; Wagenaar, E; Rijsewijk, F; Gennissen, A; Otte, A; Schuuring, E; van Ooyen, A

1990-01-01

Wnt-1 (int-1) is a cellular oncogene often activated by insertion of proviral DNA of the mouse mammary tumor virus. We have mapped the 5' end and the promoter area of the Wnt-1 gene by nuclease protection and primer extension assays. In differentiating P19 embryonal carcinoma cells, in which Wnt-1 is naturally expressed, two start sites of transcription were found, one preceded by two TATA boxes and one preceded by several GC boxes. In P19 cells, a 1-kilobase upstream sequence of Wnt-1 was able to confer differentiation-specific expression on a heterologous gene. We have investigated how Wnt-1 transcription was affected by mouse mammary tumor virus proviral integrations in various configurations near the promoters of the gene. One provirus has been inserted in the 5' nontranslated part of Wnt-1, in the same transcriptional orientation, and has functionally replaced the Wnt-1 promoters. Wnt-1 transcription in this tumor starts in the right long terminal repeat of the provirus, with considerable readthrough transcription from the left long terminal repeat. Another provirus has been inserted in the orientation opposite that of Wnt-1 into a GC box, disrupting the first Wnt-1 transcription start site but not the downstream start site. Most insertions have not structurally altered the Wnt-1 transcripts and have enhanced the activity of the normal two promoters. Images PMID:1695322

Haploinsufficiency for Translation Elongation Factor eEF1A2 in Aged Mouse Muscle and Neurons Is Compatible with Normal Function

PubMed Central

Griffiths, Lowri A.; Doig, Jennifer; Churchhouse, Antonia M. D.; Davies, Faith C. J.; Squires, Charlotte E.; Newbery, Helen J.; Abbott, Catherine M.

2012-01-01

Translation elongation factor isoform eEF1A2 is expressed in muscle and neurons. Deletion of eEF1A2 in mice gives rise to the neurodegenerative phenotype “wasted” (wst). Mice homozygous for the wasted mutation die of muscle wasting and neurodegeneration at four weeks post-natal. Although the mutation is said to be recessive, aged heterozygous mice have never been examined in detail; a number of other mouse models of motor neuron degeneration have recently been shown to have similar, albeit less severe, phenotypic abnormalities in the heterozygous state. We therefore examined the effects of ageing on a cohort of heterozygous +/wst mice and control mice, in order to establish whether a presumed 50% reduction in eEF1A2 expression was compatible with normal function. We evaluated the grip strength assay as a way of distinguishing between wasted and wild-type mice at 3–4 weeks, and then performed the same assay in older +/wst and wild-type mice. We also used rotarod performance and immunohistochemistry of spinal cord sections to evaluate the phenotype of aged heterozygous mice. Heterozygous mutant mice showed no deficit in neuromuscular function or signs of spinal cord pathology, in spite of the low levels of eEF1A2. PMID:22848658

Anaerobic digestion of tuna waste for the production of volatile fatty acids.

PubMed

Bermúdez-Penabad, Noela; Kennes, Christian; Veiga, Maria C

2017-10-01

Fish canning industries generate a significant amount of solid waste that can be digested anaerobically into volatile fatty acids (VFA). The aim of this research was to study the effect of various pHs, ranging from 5.0 to 10.0, and percentage of total solids on the anaerobic digestion of tuna waste into VFA, both in batch assays and continuous reactor. The production of VFA was affected by pH and was significantly higher under alkaline conditions. At pH 8.0, the VFA production reached 30,611mgCOD/L. The VFA mainly consisted of acetic, propionic, n-butyric and i-valeric acids. Acetic acid was the main product at all the pHs tested. In terms of total solids (TS) the best results were obtained with 2.5% total solids, reaching 0.73gCOD VFA /gCOD waste . At higher TS concentrations (5 and 8% TS) lower yields were reached probably due to inhibition at high VFA concentration. Copyright © 2017 Elsevier Ltd. All rights reserved.

Risk Reduction and Training using Simulation Based Tools - 12180

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hall, Irin P.

2012-07-01

Process Modeling and Simulation (M and S) has been used for many years in manufacturing and similar domains, as part of an industrial engineer's tool box. Traditionally, however, this technique has been employed in small, isolated projects where models were created from scratch, often making it time and cost prohibitive. Newport News Shipbuilding (NNS) has recognized the value of this predictive technique and what it offers in terms of risk reduction, cost avoidance and on-schedule performance of highly complex work. To facilitate implementation, NNS has been maturing a process and the software to rapidly deploy and reuse M and Smore » based decision support tools in a variety of environments. Some examples of successful applications by NNS of this technique in the nuclear domain are a reactor refueling simulation based tool, a fuel handling facility simulation based tool and a tool for dynamic radiation exposure tracking. The next generation of M and S applications include expanding simulation based tools into immersive and interactive training. The applications discussed here take a tool box approach to creating simulation based decision support tools for maximum utility and return on investment. This approach involves creating a collection of simulation tools that can be used individually or integrated together for a larger application. The refueling simulation integrates with the fuel handling facility simulation to understand every aspect and dependency of the fuel handling evolutions. This approach translates nicely to other complex domains where real system experimentation is not feasible, such as nuclear fuel lifecycle and waste management. Similar concepts can also be applied to different types of simulation techniques. For example, a process simulation of liquid waste operations may be useful to streamline and plan operations, while a chemical model of the liquid waste composition is an important tool for making decisions with respect to waste disposition. Integrating these tools into a larger virtual system provides a tool for making larger strategic decisions. The key to integrating and creating these virtual environments is the software and the process used to build them. Although important steps in the direction of using simulation based tools for nuclear domain, the applications described here represent only a small cross section of possible benefits. The next generation of applications will, likely, focus on situational awareness and adaptive planning. Situational awareness refers to the ability to visualize in real time the state of operations. Some useful tools in this area are Geographic Information Systems (GIS), which help monitor and analyze geographically referenced information. Combined with such situational awareness capability, simulation tools can serve as the platform for adaptive planning tools. These are the tools that allow the decision maker to react to the changing environment in real time by synthesizing massive amounts of data into easily understood information. For the nuclear domains, this may mean creation of Virtual Nuclear Systems, from Virtual Waste Processing Plants to Virtual Nuclear Reactors. (authors)« less

Determination of DNA Binding Behavior of FoxA1 Constructs Using a Gold Nanoparticle-Based High Throughput Assay

NASA Astrophysics Data System (ADS)

Aung, Khin Moh Moh; Lim, Michelle Gek Liang; Hong, Shuzhen; Cheung, Edwin; Su, Xiaodi

Forkhead box protein 1 (FoxA1) is a member of the forkhead family of winged-helix transcription factors. It plays crucial roles in the development and differentiation of multiple organs and in the regulation of estrogen-stimulated genes. In this study, in order to determine the regions of FoxA1 necessary for efficient Deoxyribonucleic Acid (DNA) binding, we cloned, expressed and purified a series of FoxA1 constructs that contain either the DNA Binding Domain (DBD), the Transcription Activation Domain (TAD), or both. We determined the DNA binding behavior of these constructs using traditional electrophoretic mobility shift assay (EMSA) and a recently developed gold nanoparticles (AuNPs)-based fast screening method. We conclude that just the DBD region alone is not sufficient for protein-DNA binding activity. Amino acids flanking the upstream of the DBD region are required for maximal DNA binding activity. Through this study, we have also further validated the AuNPs assay for its generality and expanded the existing protocol for comparing the DNA binding behavior of multiple proteins of different charge properties and molecular weights.

Waste Soybean Oil and Corn Steep Liquor as Economic Substrates for Bioemulsifier and Biodiesel Production by Candida lipolytica UCP 0998

PubMed Central

Souza, Adriana Ferreira; Rodriguez, Dayana M.; Ribeaux, Daylin R.; Luna, Marcos A. C.; Lima e Silva, Thayse A.; Andrade, Rosileide F. Silva; Gusmão, Norma B.; Campos-Takaki, Galba M.

2016-01-01

Almost all oleaginous microorganisms are available for biodiesel production, and for the mechanism of oil accumulation, which is what makes a microbial approach economically competitive. This study investigated the potential that the yeast Candida lipolytica UCP0988, in an anamorphous state, has to produce simultaneously a bioemulsifier and to accumulate lipids using inexpensive and alternative substrates. Cultivation was carried out using waste soybean oil and corn steep liquor in accordance with 22 experimental designs with 1% inoculums (107 cells/mL). The bioemulsifier was produced in the cell-free metabolic liquid in the late exponential phase (96 h), at Assay 4 (corn steep liquor 5% and waste soybean oil 8%), with 6.704 UEA, IE24 of 96.66%, and showed an anionic profile. The emulsion formed consisted of compact small and stable droplets (size 0.2–5 µm), stable at all temperatures, at pH 2 and 4, and 2% salinity, and showed an ability to remove 93.74% of diesel oil from sand. The displacement oil (ODA) showed 45.34 cm2 of dispersion (central point of the factorial design). The biomass obtained from Assay 4 was able to accumulate lipids of 0.425 g/g biomass (corresponding to 42.5%), which consisted of Palmitic acid (28.4%), Stearic acid (7.7%), Oleic acid (42.8%), Linoleic acid (19.0%), and γ-Linolenic acid (2.1%). The results showed the ability of C. lipopytica to produce both bioemulsifier and biodiesel using the metabolic conversion of waste soybean oil and corn steep liquor, which are economic renewable sources. PMID:27669227

A two-fold reduction in measurement time for neutron assay: Initial tests of a prototype dual-gated shift register

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stewart, J.E.; Bourret, S.C.; Krick, M.S.

1996-09-01

Neutron coincidence counting (NCC) is used routinely around the world for nondestructive mass assay of uranium and plutonium in many forms, including waste. Compared with other methods, NCC is generally the most flexible, economic, and rapid. Many applications of NCC would benefit from a reduction in counting time required for a fixed random error. We have developed and tested the first prototype of a dual- gated, shift-register-based electronics unit that offers the potential of decreased measurement time for all passive and active NCC applications.

A two-fold reduction in measurement time for neutron assay: Initial tests of a prototype dual-gated shift register

DOE Office of Scientific and Technical Information (OSTI.GOV)

Stewart, J.E.; Bourret, S.C.; Krick, M.S.

1996-12-31

Neutron coincidence counting (NCC) is used routinely around the world for nondestructive mass assay of uranium and plutonium in many forms, including waste. Compared with other methods, NCC is generally the most flexible, economic, and rapid. Many applications of NCC would benefit from a reduction in counting time required for a fixed random error. The authors have developed and tested the first prototype of a dual-gated, shift-register-based electronics unit that offers the potential of decreased measurement time for all passive and active NCC applications.

Arsenic and lead contamination in urban soils of Villa de la Paz (Mexico) affected by historical mine wastes and its effect on children's health studied by micronucleated exfoliated cells assay.

PubMed

Gamiño-Gutiérrez, Sandra P; González-Pérez, C Ivonne; Gonsebatt, María E; Monroy-Fernández, Marcos G

2013-02-01

Environmental geochemical and health studies were carried out in urban areas of Villa de la Paz, S.L.P. (Mexico), where mining activities have been developed for more of 200 years, leading to the pollution of surface soil by arsenic and heavy metals (Pb, Cd, Cu, Zn). The analysis of urban soils to determine total and bioaccessibility concentrations of As and Pb, demonstrated a combined contribution of the natural and anthropogenic concentrations in the site, at levels higher than the environmental guideline values that provoke a human health risk. Contour soil mapping confirmed that historical mine waste deposits without environmental control measures, are the main source of pollution soil by As and Pb in the site. Exposure (Pb in blood and As in urine) and effect (micronucleated exfoliated cells assay) biological monitoring were then carried out in the childhood population of the site and in a control site. The exposure biological monitoring demonstrated that at least 20-30 % of children presented Pb and As exposure values higher than the national and international maximum intervention values. The effect biomonitoring by MEC assay confirmed that there is a genotoxic damage in local childhood population that could be associated with the arsenic exposure in the site.

Electrohydrolysis pretreatment for enhanced methane production from lignocellulose waste pulp and paper mill sludge and its kinetics.

PubMed

Veluchamy, C; Raju, V Wilson; Kalamdhad, Ajay S

2018-03-01

A novel electrohydrolysis pretreatment enhances methane production from lignocellulose material during anaerobic digestion. A biochemical methane potential assay was carried out to determine the effect of direct current and the efficacy of electrohydrolysis pretreatment on biogas production. Methane yield was increased by 13.8%, to 301 ± 3 mL CH 4 /g VS, when lignocellulosic waste was pretreated with electrohydrolysis. A net energy gain of 13,224 kJ was realized after electrohydrolysis pretreatment, which was 1.51 times higher than reported for thermal pretreatment. In addition, two kinetic models were used, including the modified Gompertz model to reproduce the experimental data. These finding support the potential for increased methane recovery from lignocellulosic waste using electrohydrolysis as a pretreatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Kinetic Tetrazolium Microtiter Assay

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Stowe, Raymond; Koenig, David

1993-01-01

Kinetic tetrazolium microtiter assay (KTMA) involves use of tetrazolium salts and Triton X-100 (or equivalent), nontoxic, in vitro color developer solubilizing colored metabolite formazan without injuring or killing metabolizing cells. Provides for continuous measurement of metabolism and makes possible to determine rate of action of antimicrobial agent in real time as well as determines effective inhibitory concentrations. Used to monitor growth after addition of stimulatory compounds. Provides for kinetic determination of efficacy of biocide, greatly increasing reliability and precision of results. Also used to determine relative effectiveness of antimicrobial agent as function of time. Capability of generating results on day of test extremely important in treatment of water and waste, disinfection of hospital rooms, and in pharmaceutical, agricultural, and food-processing industries. Assay also used in many aspects of cell biology.

Apparatus and method for quantitative assay of generic transuranic wastes from nuclear reactors

DOEpatents

Caldwell, J.T.; Kunz, W.E.; Atencio, J.D.

1982-03-31

A combination of passive and active neutron measurements which yields quantitative information about the isotopic composition of transuranic wastes from nuclear power or weapons material manufacture reactors is described. From the measurement of prompt and delayed neutron emission and the incidence of two coincidentally emitted neutrons from induced fission of fissile material in the sample, one can quantify /sup 233/U, /sup 235/U and /sup 239/Pu isotopes in waste samples. Passive coincidence counting, including neutron multiplicity measurement and determination of the overall passive neutron flux additionally enables the separate quantitative evaluation of spontaneous fission isotopes such as /sup 240/Pu, /sup 244/Cm and /sup 252/Cf, and the spontaneous alpha particle emitter /sup 241/Am. These seven isotopes are the most important constituents of wastes from nuclear power reactors and once the mass of each isotope present is determined by the apparatus and method of the instant invention, the overall alpha particle activity can be determined to better than 1 nCi/g from known radioactivity data. Therefore, in addition to the quantitative analysis of the waste sample useful for later reclamation purposes, the alpha particle activity can be determined to decide whether permanent low-level burial is appropriate for the waste sample.

Apparatus and method for quantitative assay of generic transuranic wastes from nuclear reactors

DOEpatents

Caldwell, John T.; Kunz, Walter E.; Atencio, James D.

1984-01-01

A combination of passive and active neutron measurements which yields quantitative information about the isotopic composition of transuranic wastes from nuclear power or weapons material manufacture reactors is described. From the measurement of prompt and delayed neutron emission and the incidence of two coincidentally emitted neutrons from induced fission of fissile material in the sample, one can quantify .sup.233 U, .sup.235 U and .sup.239 Pu isotopes in waste samples. Passive coincidence counting, including neutron multiplicity measurement and determination of the overall passive neutron flux additionally enables the separate quantitative evaluation of spontaneous fission isotopes such as .sup.240 Pu, .sup.244 Cm and .sup.252 Cf, and the spontaneous alpha particle emitter .sup.241 Am. These seven isotopes are the most important constituents of wastes from nuclear power reactors and once the mass of each isotope present is determined by the apparatus and method of the instant invention, the overall alpha particle activity can be determined to better than 1 nCi/g from known radioactivity data. Therefore, in addition to the quantitative analysis of the waste sample useful for later reclamation purposes, the alpha particle activity can be determined to decide whether "permanent" low-level burial is appropriate for the waste sample.

Co-digestion of pig slaughterhouse waste with sewage sludge.

PubMed

Borowski, Sebastian; Kubacki, Przemysław

2015-06-01

Slaughterhouse wastes (SHW) are potentially very attractive substrates for biogas production. However, mono-digestion of these wastes creates great technological problems associated with the inhibitory effects of ammonia and fatty acids on methanogens as well as with the foaming in the digesters. In the following study, the co-digestion of slaughterhouse wastes with sewage sludge (SS) was undertaken. Batch and semi-continuous experiments were performed at 35°C with municipal sewage sludge and pig SHW composed of meat tissue, intestines, bristles and post-flotation sludge. In batch assays, meat tissue and intestinal wastes gave the highest methane productions of 976 and 826 dm(3)/kg VS, respectively, whereas the methane yield from the sludge was only 370 dm(3)/kg VS. The co-digestion of sewage sludge with 50% SHW (weight basis) provided the methane yield exceeding 600 dm(3)/kg VS, which was more than twice as high as the methane production from sewage sludge alone. However, when the loading rate exceeded 4 kg VS/m(3) d, a slight inhibition of methanogenesis was observed, without affecting the digester stability. The experiments showed that the co-digestion of sewage sludge with large amount of slaughterhouse wastes is feasible, and the enhanced methane production does not affect the digester stability. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of size and thermophilic pre-hydrolysis of banana peel during anaerobic digestion, and biomethanation potential of key tropical fruit wastes.

PubMed

Odedina, Mary Jesuyemi; Charnnok, Boonya; Saritpongteeraka, Kanyarat; Chaiprapat, Sumate

2017-10-01

Methane production potential of tropical fruit wastes, namely lady-finger banana peel, rambutan waste and longan waste were compared using BMP assay and stoichiometric modified Buswell and Mueller equation. Methane yields based on volatile solid (VS) were in the order of ground banana peel, chopped banana peel, chopped longan waste, and chopped rambutan waste (330.6, 268.3, 234.6 and 193.2 mLCH 4 /gVS) that corresponded to their calculated biodegradability. In continuous operations of banana peel digestion at feed concentrations based on total solid (TS) 1-2%, mesophilic single stage digester run at 20-day hydraulic retention time (20-day HRT) failed at 2%TS, but successfully recovered at 1.5%TS. Pre-hydrolysis thermophilic reactor (4-d HRT) was placed as pre-treatment to mesophilic reactor (20-d HRT). Higher biogas (with an evolution of H 2 ) and energy yields were obtained and greater system stability was achieved over the single stage digestion, particularly at higher solid feedstock. The best performance of two stage digestion was 68.5% VS destruction and energy yield of 2510.9kJ/kgVS added at a feed concentration of 2%TS. Copyright © 2017 Elsevier Ltd. All rights reserved.

Efficiency of inductively torch plasma operating at atmospheric pressure on destruction of chlorinated liquid wastes- A path to the treatment of radioactive organic halogen liquid wastes

NASA Astrophysics Data System (ADS)

Kamgang-Youbi, G.; Poizot, K.; Lemont, F.

2012-12-01

The performance of a plasma reactor for the degradation of chlorinated hydrocarbon waste is reported. Chloroform was used as a target for a recently patented destruction process based using an inductive plasma torch. Liquid waste was directly injected axially into the argon plasma with a supplied power of ~4 kW in the presence of oxygen as oxidant and carrier gas. Decomposition was performed at CHCl3 feed rates up to 400 g·h-1 with different oxygen/waste molar ratios, chloroform destruction was obtained with at least 99% efficiency and the energy efficiency reached 100 g·kWh-1. The conversion end products were identified and assayed by online FTIR spectroscopy (CO2, HCl and H2O) and redox titration (Cl2). Considering phosgene as representative of toxic compounds, only very small quantities of toxics were released (< 1 g·h-1) even with high waste feed rates. The experimental results were very close to the equilibrium composition predicted by thermodynamic calculations. At the bottom of the reactor, the chlorinated acids were successfully trapped in a scrubber and transformed into mineral salts, hence, only CO2 and H2O have been found in the final off-gases composition.

APPLICATION, PERFORMANCE, AND COSTS OF ...

EPA Pesticide Factsheets

A critical review of biological treatment processes for remediation of contaminated soils is presented. The focus of the review is on documented cost and performance of biological treatment technologies demonstrated at full- or field-scale. Some of the data were generated by the U.S. Environmental Protection Agency's (EPA's) Bioremediation in the Field Program, jointly supported by EPA's Office of Research and Development, EPA's Office of Solid Waste and Emergency Waste, and the EPA Regions through the Superfund Innovative Technology Evaluation Program (SITE) Program. Military sites proved to be another fertile data source. Technologies reviewed in this report include both ex-situ processes, (land treatment, biopile/biocell treatment, composting, and bioslurry reactor treatment) and in-situ alternatives (conventional bioventing, enhanced or cometabolic bioventing, anaerobic bioventing, bioslurping, phytoremediation, and natural attenuation). Targeted soil contaminants at the documented sites were primarily organic chemicals, including BTEX, petroleum hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), chlorinated aliphatic hydrocarbons (CAHs), organic solvents, polychlorinated biphenyls (PCBs), pesticides, dioxin, and energetics. The advantages, limitations, and major cost drivers for each technology are discussed. Box and whisker plots are used to summarize before and after concentrations of important contaminant groups for those technologies consider

Environemental Planning Quarterly

NASA Astrophysics Data System (ADS)

Anderson, Mary P.

The Environmental Planning Quarterly is a publication of the Environmental Planning Division of the American Planning Association (APA). Its current editor is G. William Page (Department of Urban Planning, University of Wisconsin-Milwaukee). A recent issue included a feature article on forest management in the state of Washington and several lengthy and informative news items about activities of the Environmental Protection Agency related to groundwater contamination and hazardous waste. Other news items contained information on air pollution and acid rain as well as on surface water pollution. Subscriptions can be obtained by writing to the Environmental Planning Division. American Planning Association, Lock Box 97774, Chicago, IL 60690. Annual fees are as follows: $ 15 (APA member), $25 (affiliate), $5 (student), $30 (subscribing).

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chua, S.E.; Abito, G.F.

A 1-MWt multi-crop drying facility using low-enthalpy waste geothermal heat is installed within the vicinity of the Southern Negros Geothermal Project (January, 1994). The plant is envisioned to demonstrate the direct use of geothermal resources for agro-industrial purposes and at the same time, provide major benefits by raising the quality of the agro-industrial products to meet higher standards. The development and design of the heat exchangers that supply the heat and the dryer used in the facility is presented. The process flow and the dryer parameters in the drying of coconut meat and other crops have been determined. The initialmore » design of the dryers target the dehydration of coconut meat and other crops using boxes and trays.« less

Stereochemical analysis of the functional significance of the conserved inverted CCAAT and TATA elements in the rat bone sialoprotein gene promoter.

PubMed

Su, Ming; Lee, Daniel; Ganss, Bernhard; Sodek, Jaro

2006-04-14

Basal transcription of the bone sialoprotein gene is mediated by highly conserved inverted CCAAT (ICE; ATTGG) and TATA elements (TTTATA) separated by precisely 21 nucleotides. Here we studied the importance of the relative position and orientation of the CCAAT and TATA elements in the proximal promoter by measuring the transcriptional activity of a series of mutated reporter constructs in transient transfection assays. Whereas inverting the TTTATA (wild type) to a TATAAA (consensus TATA) sequence increased transcription slightly, transcription was reduced when the flanking dinucleotides were also inverted. In contrast, reversing the ATTGG (wild type; ICE) to a CCAAT (RICE) sequence caused a marked reduction in transcription, whereas both transcription and NF-Y binding were progressively increased with the simultaneous inversion of flanking nucleotides (f-RICE-f). Reducing the distance between the ICE and TATA elements produced cyclical changes in transcriptional activity that correlated with progressive alterations in the relative positions of the CCAAT and TATA elements on the face of the DNA helix. Minimal transcription was observed after 5 nucleotides were deleted (equivalent to approximately one half turn of the helix), whereas transcription was fully restored after deleting 10 nucleotides (approximately one full turn of the DNA helix), transcriptional activity being progressively lost with deletions beyond 10 nucleotides. In comparison, when deletions were made with the ICE in the reversed (f-RICE-f) orientation transcriptional activity was progressively lost with no recovery. These results show that, although transcription can still occur when the CCAAT box is reversed and/or displaced relative to the TATA box, the activity is dependent upon the flexibility of the intervening DNA helix needed to align the NF-Y complex on the CCAAT box with preinitiation complex proteins that bind to the TATA box. Thus, the precise location and orientation of the CCAAT element is necessary for optimizing basal transcription of the bone sialoprotein gene.

Loss of MYC and E-box3 binding contributes to defective MYC-mediated transcriptional suppression of human MC-let-7a-1~let-7d in glioblastoma

PubMed Central

Wang, Zifeng; Lin, Sheng; Zhang, Ji; Xu, Zhenhua; Xiang, Yu; Yao, Hong; Ge, Lei; Xie, Dan; Kung, Hsiang-fu; Lu, Gang; Poon, Wai Sang; Liu, Quentin; Lin, Marie Chia-mi

2016-01-01

Previously, we reported that MYC oncoprotein down-regulates the transcription of human MC-let-7a-1~let-7d microRNA cluster in hepatocarcinoma (HCC). Surprisingly, in silico analysis indicated that let-7 miRNA expression levels are not reduced in glioblastoma (GBM). Here we investigated the molecular basis of this differential expression. Using human GBM U87 and U251 cells, we first demonstrated that forced over-expression of MYC indeed could not down-regulate the expression of human MC-let-7a-1~let-7d microRNA cluster in GBM. Furthermore, analysis of MC-let-7a-1~let-7d promoter in GBM indicated that MYC failed to inhibit the promoter activity. Pearson's correlation and Linear Regression analysis using the expression data from GSE55092 (HCC) and GSE4290 (GBM) demonstrated a converse relationship of MC-let-7a-1~let-7d and MYC only in HCC but not in GBM. To understand the underlying mechanisms, we examined whether MYC could bind to the non-canonical E-box 3 located in the promoter of MC-let-7a-1~let-7d. Results from both chromatin immune-precipitation (ChIP) and super-shift assays clearly demonstrated the loss of MYC and E-box 3 binding in GBM, suggesting for the first time that a defective MYC and E-box3 binding in GBM is responsible for the differential MYC mediated transcriptional inhibition of MC-let-7a-1~let-7d and potentially other tumor suppressors. MYC and let-7 are key oncoprotein and tumor suppressor, respectively. Understanding the molecular mechanisms of their regulations will provide new insight and have important implications in the therapeutics of GBM as well as other cancers. PMID:27409345

Binding Sequences for RdgB, a DNA Damage-Responsive Transcriptional Activator, and Temperature-Dependent Expression of Bacteriocin and Pectin Lyase Genes in Pectobacterium carotovorum subsp. carotovorum▿ †

PubMed Central

Yamada, Kazuteru; Kaneko, Jun; Kamio, Yoshiyuki; Itoh, Yoshifumi

2008-01-01

Pectobacterium carotovorum subsp. carotovorum strain Er simultaneously produces the phage tail-like bacteriocin carotovoricin (Ctv) and pectin lyase (Pnl) in response to DNA-damaging agents. The regulatory protein RdgB of the Mor/C family of proteins activates transcription of pnl through binding to the promoter. However, the optimal temperature for the synthesis of Ctv (23°C) differs from that for synthesis of Pnl (30°C), raising the question of whether RdgB directly activates ctv transcription. Here we report that RdgB directly regulates Ctv synthesis. Gel mobility shift assays demonstrated RdgB binding to the P0, P1, and P2 promoters of the ctv operons, and DNase I footprinting determined RdgB-binding sequences (RdgB boxes) on these and on the pnl promoters. The RdgB box of the pnl promoter included a perfect 7-bp inverted repeat with high binding affinity to the regulator (Kd [dissociation constant] = 150 nM). In contrast, RdgB boxes of the ctv promoters contained an imperfect inverted repeat with two or three mismatches that consequently reduced binding affinity (Kd = 250 to 350 nM). Transcription of the rdgB and ctv genes was about doubled at 23°C compared with that at 30°C. In contrast, the amount of pnl transcription tripled at 30°C. Thus, the inverse synthesis of Ctv and Pnl as a function of temperature is apparently controlled at the transcriptional level, and reduced rdgB expression at 30°C obviously affected transcription from the ctv promoters with low-affinity RdgB boxes. Pathogenicity toward potato tubers was reduced in an rdgB knockout mutant, suggesting that the RdgAB system contributes to the pathogenicity of this bacterium, probably by activating pnl expression. PMID:18689515

Ciliary dyslexia candidate genes DYX1C1 and DCDC2 are regulated by Regulatory Factor X (RFX) transcription factors through X-box promoter motifs

PubMed Central

Tammimies, Kristiina; Bieder, Andrea; Lauter, Gilbert; Sugiaman-Trapman, Debora; Torchet, Rachel; Hokkanen, Marie-Estelle; Burghoorn, Jan; Castrén, Eero; Kere, Juha; Tapia-Páez, Isabel; Swoboda, Peter

2016-01-01

DYX1C1, DCDC2, and KIAA0319 are three of the most replicated dyslexia candidate genes (DCGs). Recently, these DCGs were implicated in functions at the cilium. Here, we investigate the regulation of these DCGs by Regulatory Factor X transcription factors (RFX TFs), a gene family known for transcriptionally regulating ciliary genes. We identify conserved X-box motifs in the promoter regions of DYX1C1, DCDC2, and KIAA0319 and demonstrate their functionality, as well as the ability to recruit RFX TFs using reporter gene and electrophoretic mobility shift assays. Furthermore, we uncover a complex regulation pattern between RFX1, RFX2, and RFX3 and their significant effect on modifying the endogenous expression of DYX1C1 and DCDC2 in a human retinal pigmented epithelial cell line immortalized with hTERT (hTERT-RPE1). In addition, induction of ciliogenesis increases the expression of RFX TFs and DCGs. At the protein level, we show that endogenous DYX1C1 localizes to the base of the cilium, whereas DCDC2 localizes along the entire axoneme of the cilium, thereby validating earlier localization studies using overexpression models. Our results corroborate the emerging role of DCGs in ciliary function and characterize functional noncoding elements, X-box promoter motifs, in DCG promoter regions, which thus can be targeted for mutation screening in dyslexia and ciliopathies associated with these genes.—Tammimies, K., Bieder, A., Lauter, G., Sugiaman-Trapman, D., Torchet, R., Hokkanen, M.-E., Burghoorn, J., Castrén, E., Kere, J., Tapia-Páez, I., Swoboda, P. Ciliary dyslexia candidate genes DYX1C1 and DCDC2 are regulated by Regulatory Factor (RF) X transcription factors through X-box promoter motifs. PMID:27451412

Ciliary dyslexia candidate genes DYX1C1 and DCDC2 are regulated by Regulatory Factor X (RFX) transcription factors through X-box promoter motifs.

PubMed

Tammimies, Kristiina; Bieder, Andrea; Lauter, Gilbert; Sugiaman-Trapman, Debora; Torchet, Rachel; Hokkanen, Marie-Estelle; Burghoorn, Jan; Castrén, Eero; Kere, Juha; Tapia-Páez, Isabel; Swoboda, Peter

2016-10-01

DYX1C1, DCDC2, and KIAA0319 are three of the most replicated dyslexia candidate genes (DCGs). Recently, these DCGs were implicated in functions at the cilium. Here, we investigate the regulation of these DCGs by Regulatory Factor X transcription factors (RFX TFs), a gene family known for transcriptionally regulating ciliary genes. We identify conserved X-box motifs in the promoter regions of DYX1C1, DCDC2, and KIAA0319 and demonstrate their functionality, as well as the ability to recruit RFX TFs using reporter gene and electrophoretic mobility shift assays. Furthermore, we uncover a complex regulation pattern between RFX1, RFX2, and RFX3 and their significant effect on modifying the endogenous expression of DYX1C1 and DCDC2 in a human retinal pigmented epithelial cell line immortalized with hTERT (hTERT-RPE1). In addition, induction of ciliogenesis increases the expression of RFX TFs and DCGs. At the protein level, we show that endogenous DYX1C1 localizes to the base of the cilium, whereas DCDC2 localizes along the entire axoneme of the cilium, thereby validating earlier localization studies using overexpression models. Our results corroborate the emerging role of DCGs in ciliary function and characterize functional noncoding elements, X-box promoter motifs, in DCG promoter regions, which thus can be targeted for mutation screening in dyslexia and ciliopathies associated with these genes.-Tammimies, K., Bieder, A., Lauter, G., Sugiaman-Trapman, D., Torchet, R., Hokkanen, M.-E., Burghoorn, J., Castrén, E., Kere, J., Tapia-Páez, I., Swoboda, P. Ciliary dyslexia candidate genes DYX1C1 and DCDC2 are regulated by Regulatory Factor (RF) X transcription factors through X-box promoter motifs. © The Author(s).

Rice phytochrome-interacting factor protein OsPIF14 represses OsDREB1B gene expression through an extended N-box and interacts preferentially with the active form of phytochrome B

DOE PAGES

Cordeiro, André M.; Figueiredo, Duarte D.; Tepperman, James; ...

2015-12-28

DREB1/CBF genes, known as major regulators of plant stress responses, are rapidly and transiently induced by low temperatures. Using a yeast one-hybrid screening, we identified a putative Phytochrome-Interacting bHLH Factor (OsPIF14), as binding to the OsDREB1B promoter. bHLH proteins are able to bind to hexameric E-box (CANNTG) or N-box (CACG(A/C)G) motifs, depending on transcriptional activity. We have shown that OsPIF14 binds to the OsDREB1B promoter through two N-boxes and that the flanking regions of the hexameric core are essential for protein–DNA interaction and stability. We also showed that OsPIF14 down-regulates OsDREB1B gene expression in rice protoplasts, corroborating the OsPIF14 repressormore » activity observed in the transactivation assays using Arabidopsis protoplasts. Additionally, we showed that OsPIF14 is indeed a phytochrome interacting factor, which preferentially binds to the active form (Pfr) of rice phytochrome B. This raises the possibility that OsPIF14 activity might be modulated by light. However, we did not observe any regulation of the OsDREB1B gene expression by light under control conditions. Moreover, OsPIF14 gene expression was shown to be modulated by different treatments, such as drought, salt, cold and ABA. Interestingly, OsPIF14 showed also a specific cold-induced alternative splicing. Our results suggest the possibility that OsPIF14 is involved in cross-talk between light and stress signaling through interaction with the OsDREB1B promoter. Finally, although in the absence of stress, OsDREB1B gene expression was not regulated by light, given previous reports, it remains possible that OsPIF14 has a role in light modulation of stress responses.« less

Rice phytochrome-interacting factor protein OsPIF14 represses OsDREB1B gene expression through an extended N-box and interacts preferentially with the active form of Phytochrome B

PubMed Central

Cordeiro, André M.; Figueiredo, Duarte D.; Tepperman, James; Borba, Ana Rita; Lourenço, Tiago; Abreu, Isabel A.; Ouwerkerk, Pieter B.F.; Quail, Peter H.; Oliveira, M. Margarida; Saibo, Nelson J. M.

2016-01-01

DREB1/CBF genes, known as major regulators of plant stress responses, are rapidly and transiently induced by low temperatures. Using a Yeast one Hybrid screening, we identified a putative Phytochrome-Interacting bHLH Factor (OsPIF14), as binding to the OsDREB1B promoter. bHLH proteins are able to bind to hexameric E-box (CANNTG) or N-box (CACG(A/C)G) motifs, depending on transcriptional activity. We have shown that OsPIF14 binds to the OsDREB1B promoter through two N-boxes and that the flanking regions of the hexameric core are essential for protein-DNA interaction and stability. We also showed that OsPIF14 down-regulates OsDREB1B gene expression in rice protoplasts, corroborating the OsPIF14 repressor activity observed in the transactivation assays using Arabidopsis protoplasts. In addition, we showed that OsPIF14 is indeed a Phytochrome Interacting Factor, which preferentially binds to the active form (Pfr) of rice phytochrome B. This raises the possibility that OsPIF14 activity might be modulated by light. However, we did not observe any regulation of the OsDREB1B gene expression by light under control conditions. Moreover, OsPIF14 gene expression was shown to be modulated by different treatments, such as drought, salt, cold and ABA. Interestingly, OsPIF14 showed also a specific cold-induced alternative splicing. All together, these results suggest the possibility that OsPIF14 is involved in cross-talk between light and stress signaling through interaction with the OsDREB1B promoter. Although in the absence of stress, OsDREB1B gene expression was not regulated by light, given previous reports, it remains possible that OsPIF14 has a role in light modulation of stress responses. PMID:26732823

Urinary heavy metal levels and relevant factors among people exposed to e-waste dismantling.

PubMed

Wang, Hongmei; Han, Mei; Yang, Suwen; Chen, Yanqing; Liu, Qian; Ke, Shen

2011-01-01

Primitive electronic waste (e-waste) recycling has become a growing environmental concern, and toxic heavy metals released from e-waste activities may continue to threaten the health of local people. To study the impact of heavy metals in people around e-waste sites, 349 people from e-waste recycling sites (exposure group) and 118 people from a green plantation (control group) were surveyed, and their urinary levels of lead (UPb), cadmium (UCd), manganese (UMn), copper (UCu), and Zinc (UZn) were assayed. Questionnaire surveys for risk factors were also performed and analyzed by using the Pearson correlation analysis. Results indicated that the levels of urinary Cd in both occupational dismantling people {GM(GSD) 0.72(0.71) ug/L} and non-occupational dismantling people {GM(GSD) 0.50(0.79) ug/L} were higher than the control group {GM(GSD) 0.27(0.85) ug/L}. Further analyses of correlations between urinary heavy metal levels and exposure factors in the exposed group revealed positive relationship between the duration of dismantling and the level of UPb (p < 0.05). Meanwhile, rice sources from local village have a positive correlation with the level of UPb and UCd (p < 0.01). Other factors, however, may also have influences on heavy metal burden, and not all urinary heavy metal levels can be contributed to e-waste dismantling exposure levels. Primitive e-waste recycling activities may contribute to the changes of urinary heavy metal levels and increase the health risk for those chronically working on e-waste dismantling. Copyright © 2010 Elsevier Ltd. All rights reserved.

Duplex unwinding and ATPase activities of the DEAD-box helicase eIF4A are coupled by eIF4G and eIF4B

PubMed Central

Özeş, Ali R.; Feoktistova, Kateryna; Avanzino, Brian C.; Fraser, Christopher S.

2011-01-01

Eukaryotic initiation factor 4A (eIF4A) is a DEAD-box helicase that stimulates translation initiation by unwinding mRNA secondary structure. The accessory proteins, eIF4G, eIF4B, and eIF4H enhance the duplex unwinding activity of eIF4A, but the extent to which they modulate eIF4A activity is poorly understood. Here, we use real time fluorescence assays to determine the kinetic parameters of duplex unwinding and ATP hydrolysis by these initiation factors. To ensure efficient duplex unwinding, eIF4B and eIF4G cooperatively activate the duplex unwinding activity of eIF4A. Our data reveal that eIF4H is much less efficient at stimulating eIF4A unwinding activity than eIF4B, implying that eIF4H is not able to completely substitute for eIF4B in duplex unwinding. By monitoring unwinding and ATPase assays using identical conditions, we demonstrate that eIF4B couples the ATP hydrolysis cycle of eIF4A with strand separation, thereby minimizing non-productive unwinding events. Using duplex substrates with altered GC contents, but with similar predicted thermal stabilities, we further show that the rate of formation of productive unwinding complexes is strongly influenced by the local stability per base pair in addition to the stability of the entire duplex. This finding explains how a change in the GC content of a hairpin while maintaining overall predicted thermal stability is able to influence translation initiation. PMID:21840318

The pea END1 promoter drives anther-specific gene expression in different plant species.

PubMed

Gómez, María D; Beltrán, José-Pío; Cañas, Luis A

2004-10-01

END1 was isolated by an immunosubtractive approach intended to identify specific proteins present in the different pea (Pisum sativum L.) floral organs and the genes encoding them. Following this strategy we obtained a monoclonal antibody (mAbA1) that specifically recognized a 26-kDa protein (END1) only detected in anther tissues. Northern blot assays showed that END1 is expressed specifically in the anther. In situ hybridization and immunolocalization assays corroborated the specific expression of END1 in the epidermis, connective, endothecium and middle layer cells during the different stages of anther development. END1 is the first anther-specific gene isolated from pea. The absence of a practicable pea transformation method together with the fact that no END1 homologue gene exists in Arabidopsis prevented us from carrying out END1 functional studies. However, we designed functional studies with the END1 promoter in different dicot species, as the specific spatial and temporal expression pattern of END1 suggested, among other things, the possibility of using its promoter region for biotechnological applications. Using different constructs to drive the uidA (beta-glucuronidase) gene controlled by the 2.7-kb isolated promoter sequence we have proven that the END1 promoter is fully functional in the anthers of transgenic Arabidopsis thaliana (L.) Heynh., Nicotiana tabacum L. (tobacco) and Lycopersicon esculentum Mill. (tomato) plants. The presence in the -330-bp region of the promoter sequence of three putative CArG boxes also suggests that END1 could be a target gene of MADS-box proteins and that, subsequently, it would be activated by genes controlling floral organ identity.

Interactions between the S-domain receptor kinases and AtPUB-ARM E3 ubiquitin ligases suggest a conserved signaling pathway in Arabidopsis.

PubMed

Samuel, Marcus A; Mudgil, Yashwanti; Salt, Jennifer N; Delmas, Frédéric; Ramachandran, Shaliny; Chilelli, Andrea; Goring, Daphne R

2008-08-01

The Arabidopsis (Arabidopsis thaliana) genome encompasses multiple receptor kinase families with highly variable extracellular domains. Despite their large numbers, the various ligands and the downstream interacting partners for these kinases have been deciphered only for a few members. One such member, the S-receptor kinase, is known to mediate the self-incompatibility (SI) response in Brassica. S-receptor kinase has been shown to interact and phosphorylate a U-box/ARM-repeat-containing E3 ligase, ARC1, which, in turn, acts as a positive regulator of the SI response. In an effort to identify conserved signaling pathways in Arabidopsis, we performed yeast two-hybrid analyses of various S-domain receptor kinase family members with representative Arabidopsis plant U-box/ARM-repeat (AtPUB-ARM) E3 ligases. The kinase domains from S-domain receptor kinases were found to interact with ARM-repeat domains from AtPUB-ARM proteins. These kinase domains, along with M-locus protein kinase, a positive regulator of SI response, were also able to phosphorylate the ARM-repeat domains in in vitro phosphorylation assays. Subcellular localization patterns were investigated using transient expression assays in tobacco (Nicotiana tabacum) BY-2 cells and changes were detected in the presence of interacting kinases. Finally, potential links to the involvement of these interacting modules to the hormone abscisic acid (ABA) were investigated. Interestingly, AtPUB9 displayed redistribution to the plasma membrane of BY-2 cells when either treated with ABA or coexpressed with the active kinase domain of ARK1. As well, T-DNA insertion mutants for ARK1 and AtPUB9 lines were altered in their ABA sensitivity during germination and acted at or upstream of ABI3, indicating potential involvement of these proteins in ABA responses.

ArgR of Streptomyces coelicolor Is a Pleiotropic Transcriptional Regulator: Effect on the Transcriptome, Antibiotic Production, and Differentiation in Liquid Cultures

PubMed Central

Botas, Alma; Pérez-Redondo, Rosario; Rodríguez-García, Antonio; Álvarez-Álvarez, Rubén; Yagüe, Paula; Manteca, Angel; Liras, Paloma

2018-01-01

ArgR is a well-characterized transcriptional repressor controlling the expression of arginine and pyrimidine biosynthetic genes in bacteria. In this work, the biological role of Streptomyces coelicolor ArgR was analyzed by comparing the transcriptomes of S. coelicolor ΔargR and its parental strain, S. coelicolor M145, at five different times over a 66-h period. The effect of S. coelicolor ArgR was more widespread than that of the orthologous protein of Escherichia coli, affecting the expression of 1544 genes along the microarray time series. This S. coelicolor regulator repressed the expression of arginine and pyrimidine biosynthetic genes, but it also modulated the expression of genes not previously described to be regulated by ArgR: genes involved in nitrogen metabolism and nitrate utilization; the act, red, and cpk genes for antibiotic production; genes for the synthesis of the osmotic stress protector ectoine; genes related to hydrophobic cover formation and sporulation (chaplins, rodlins, ramR, and whi genes); all the cwg genes encoding proteins for glycan cell wall biosynthesis; and genes involved in gas vesicle formation. Many of these genes contain ARG boxes for ArgR binding. ArgR binding to seven new ARG boxes, located upstream or near the ectA-ectB, afsS, afsR, glnR, and redH genes, was tested by DNA band-shift assays. These data and those of previously assayed fragments permitted the construction of an improved model of the ArgR binding site. Interestingly, the overexpression of sporulation genes observed in the ΔargR mutant in our culture conditions correlated with a sporulation-like process, an uncommon phenotype. PMID:29545785

The human phospholamban gene: structure and expression.

PubMed

McTiernan, C F; Frye, C S; Lemster, B H; Kinder, E A; Ogletree-Hughes, M L; Moravec, C S; Feldman, A M

1999-03-01

Phospholamban, through modulation of sarcoplasmic reticulum calcium-ATPase activity, is a key regulator of cardiac diastolic function. Alterations in phospholamban expression may define parameters of muscle relaxation. In experimental animals, phospholamban is differentially expressed in various striated and smooth muscles, and within the four chambers of the heart. Decreased phospholamban expression within the heart during heart failure has also been observed. Furthermore, regulatory elements of mammalian phospholamban genes remain poorly defined. To extend these studies to humans, we (1) characterized phospholamban expression in various human organs, (2) isolated genomic clones encoding the human phospholamban gene, and (3) prepared human phospholamban promoter/luciferase reporter constructs and performed transient transfection assays to begin identification of regulatory elements. We observed that human ventricle and quadriceps displayed high levels of phospholamban transcripts and proteins, with markedly lower expression observed in smooth muscles, while the right atria also expressed low levels of phospholamban. The human phospholamban gene structure closely resembles that reported for chicken, rabbit, rat, and mouse. Comparison of the human to other mammalian phospholamban genes indicates a marked conservation of sequence for at least 217 bp upstream of the transcription start site, which contains conserved motifs for GATA, CP1/NFY, M-CAT-like, and E-box elements. Transient transfection assays with a series of plasmids containing deleted 5' flanking regions (between -2530 and -66 through +85) showed that sequences between -169 and the CP1-box at -93 were required for maximal promoter activity in neonatal rat cardiomyocytes. Activity of these reporters in HeLa cells was markedly lower than that observed in rat cardiomyocytes, suggesting at least a partial tissue selectivity of these reporter constructs.

Optimization of Physical Conditions for the Aqueous Extraction of Antioxidant Compounds from Ginger (Zingiber officinale) Applying a Box-Behnken Design.

PubMed

Ramírez-Godínez, Juan; Jaimez-Ordaz, Judith; Castañeda-Ovando, Araceli; Añorve-Morga, Javier; Salazar-Pereda, Verónica; González-Olivares, Luis Guillermo; Contreras-López, Elizabeth

2017-03-01

Since ancient times, ginger (Zingiber officinale) has been widely used for culinary and medicinal purposes. This rhizome possesses several chemical constituents; most of them present antioxidant capacity due mainly to the presence of phenolic compounds. Thus, the physical conditions for the optimal extraction of antioxidant components of ginger were investigated by applying a Box-Behnken experimental design. Extracts of ginger were prepared using water as solvent in a conventional solid-liquid extraction. The analyzed variables were time (5, 15 and 25 min), temperature (20, 55 and 90 °C) and sample concentration (2, 6 and 10 %). The antioxidant activity was measured using the 2,2-diphenyl-1-picrylhydrazyl method and a modified ferric reducing antioxidant power assay while total phenolics were measured by Folin & Ciocalteu's method. The suggested experimental design allowed the acquisition of aqueous extracts of ginger with diverse antioxidant activity (100-555 mg Trolox/100 g, 147-1237 mg Fe 2+ /100 g and 50-332 mg gallic acid/100 g). Temperature was the determining factor in the extraction of components with antioxidant activity, regardless of time and sample quantity. The optimal physical conditions that allowed the highest antioxidant activity were: 90 °C, 15 min and 2 % of the sample. The correlation value between the antioxidant activity by ferric reducing antioxidant power assay and the content of total phenolics was R 2  = 0.83. The experimental design applied allowed the determination of the physical conditions under which ginger aqueous extracts liberate compounds with antioxidant activity. Most of them are of the phenolic type as it was demonstrated through the correlation established between different methods used to measure antioxidant capacity.

Genotoxic and mutagenic effects of polluted surface water in the midwestern region of Brazil using animal and plant bioassays

PubMed Central

Dourado, Priscila Leocádia Rosa; da Rocha, Monyque Palagano; Roveda, Liriana Mara; Raposo, Jorge Luiz; Cândido, Liliam Sílvia; Cardoso, Claudia Andréa Lima; Morales, Maria Aparecida Marin; de Oliveira, Kelly Mari Pires; Grisolia, Alexeia Barufatti

2016-01-01

Abstract This study aimed to evaluate DNA damage in animal and plant cells exposed to water from the Água Boa stream (Dourados, Mato Grosso do Sul, Brazil) by using bioassays, and to identify the chemical compounds in the water to determine the water quality in the area. Through the cytotoxicity bioassay with Allium cepa, using micronucleus test, and comet assay, using Astyanax altiparanae fish, the results indicated that biological samples were genetically altered. Micronuclei were observed in erythrocytes of A. altiparanae after exposure to water from locations close to industrial waste discharge. The highest DNA damage observed with the comet assay in fish occurred with the exposure to water from locations where the presence of metals (Cu, Pb, Cd, Ni) was high, indicating the possibility of genotoxic effects of these compounds. Thus, these results reinforce the importance of conducting genotoxicity tests for developing management plans to improve water quality, and indicate the need for waste management before domestic and industrial effluents are released into the rivers and streams. PMID:27801481

Carotenoids, Phenolic Compounds and Tocopherols Contribute to the Antioxidative Properties of Some Microalgae Species Grown on Industrial Wastewater.

PubMed

Safafar, Hamed; van Wagenen, Jonathan; Møller, Per; Jacobsen, Charlotte

2015-12-11

This study aimed at investigating the potential of microalgae species grown on industrial waste water as a new source of natural antioxidants. Six microalgae from different classes, including Phaeodactylum sp. (Bacillariophyceae), Nannochloropsis sp. (Eustigmatophyceae), Chlorella sp., Dunaniella sp., and Desmodesmus sp. (Chlorophyta), were screened for their antioxidant properties using different in vitro assays. Natural antioxidants, including pigments, phenolics, and tocopherols, were measured in methanolic extracts of microalgae biomass. Highest and lowest concentrations of pigments, phenolic compounds, and tocopherols were found in Desmodesmus sp. and Phaeodactylum tricornuotom microalgae species, respectively. The results of each assay were correlated to the content of natural antioxidants in microalgae biomass. Phenolic compounds were found as major contributors to the antioxidant activity in all antioxidant tests while carotenoids were found to contribute to the 1,1-diphenyl-2-picryl-hydrazil (DPPH) radical scavenging activity, ferrous reduction power (FRAP), and ABTS-radical scavenging capacity activity. Desmodesmus sp. biomass represented a potentially rich source of natural antioxidants, such as carotenoids (lutein), tocopherols, and phenolic compounds when cultivated on industrial waste water as the main nutrient source.

Carotenoids, Phenolic Compounds and Tocopherols Contribute to the Antioxidative Properties of Some Microalgae Species Grown on Industrial Wastewater

PubMed Central

Safafar, Hamed; van Wagenen, Jonathan; Møller, Per; Jacobsen, Charlotte

2015-01-01

This study aimed at investigating the potential of microalgae species grown on industrial waste water as a new source of natural antioxidants. Six microalgae from different classes, including Phaeodactylum sp. (Bacillariophyceae), Nannochloropsis sp. (Eustigmatophyceae), Chlorella sp., Dunaniella sp., and Desmodesmus sp. (Chlorophyta), were screened for their antioxidant properties using different in vitro assays. Natural antioxidants, including pigments, phenolics, and tocopherols, were measured in methanolic extracts of microalgae biomass. Highest and lowest concentrations of pigments, phenolic compounds, and tocopherols were found in Desmodesmus sp. and Phaeodactylum tricornuotom microalgae species, respectively. The results of each assay were correlated to the content of natural antioxidants in microalgae biomass. Phenolic compounds were found as major contributors to the antioxidant activity in all antioxidant tests while carotenoids were found to contribute to the 1,1-diphenyl-2-picryl-hydrazil (DPPH) radical scavenging activity, ferrous reduction power (FRAP), and ABTS-radical scavenging capacity activity. Desmodesmus sp. biomass represented a potentially rich source of natural antioxidants, such as carotenoids (lutein), tocopherols, and phenolic compounds when cultivated on industrial waste water as the main nutrient source. PMID:26690454

Prion-Seeding Activity in Cerebrospinal Fluid of Deer with Chronic Wasting Disease

PubMed Central

Haley, Nicholas J.; Van de Motter, Alexandra; Carver, Scott; Henderson, Davin; Davenport, Kristen; Seelig, Davis M.; Mathiason, Candace; Hoover, Edward

2013-01-01

Transmissible spongiform encephalopathies (TSEs), or prion diseases, are a uniformly fatal family of neurodegenerative diseases in mammals that includes chronic wasting disease (CWD) of cervids. The early and ante-mortem identification of TSE-infected individuals using conventional western blotting or immunohistochemistry (IHC) has proven difficult, as the levels of infectious prions in readily obtainable samples, including blood and bodily fluids, are typically beyond the limits of detection. The development of amplification-based seeding assays has been instrumental in the detection of low levels of infectious prions in clinical samples. In the present study, we evaluated the cerebrospinal fluid (CSF) of CWD-exposed (n=44) and naïve (n=4) deer (n=48 total) for CWD prions (PrPd) using two amplification assays: serial protein misfolding cyclic amplification with polytetrafluoroethylene beads (sPMCAb) and real-time quaking induced conversion (RT-QuIC) employing a truncated Syrian hamster recombinant protein substrate. Samples were evaluated blindly in parallel with appropriate positive and negative controls. Results from amplification assays were compared to one another and to obex immunohistochemistry, and were correlated to available clinical histories including CWD inoculum source (e.g. saliva, blood), genotype, survival period, and duration of clinical signs. We found that both sPMCAb and RT-QuIC were capable of amplifying CWD prions from cervid CSF, and results correlated well with one another. Prion seeding activity in either assay was observed in approximately 50% of deer with PrPd detected by IHC in the obex region of the brain. Important predictors of amplification included duration of clinical signs and time of first tonsil biopsy positive results, and ultimately the levels of PrPd identified in the obex by IHC. Based on our findings, we expect that both sPMCAb and RT-QuIC may prove to be useful detection assays for the detection of prions in CSF. PMID:24282599

Are In Vitro Methods for the Detection of Endocrine Potentials in the Aquatic Environment Predictive for In Vivo Effects? Outcomes of the Projects SchussenAktiv and SchussenAktivplus in the Lake Constance Area, Germany

PubMed Central

Henneberg, Anja; Bender, Katrin; Blaha, Ludek; Giebner, Sabrina; Kuch, Bertram; Köhler, Heinz-R.; Maier, Diana; Oehlmann, Jörg; Richter, Doreen; Scheurer, Marco; Schulte-Oehlmann, Ulrike; Sieratowicz, Agnes; Ziebart, Simone; Triebskorn, Rita

2014-01-01

Many studies about endocrine pollution in the aquatic environment reveal changes in the reproduction system of biota. We analysed endocrine activities in two rivers in Southern Germany using three approaches: (1) chemical analyses, (2) in vitro bioassays, and (3) in vivo investigations in fish and snails. Chemical analyses were based on gas chromatography coupled with mass spectrometry. For in vitro analyses of endocrine potentials in water, sediment, and waste water samples, we used the E-screen assay (human breast cancer cells MCF-7) and reporter gene assays (human cell line HeLa-9903 and MDA-kb2). In addition, we performed reproduction tests with the freshwater mudsnail Potamopyrgus antipodarum to analyse water and sediment samples. We exposed juvenile brown trout (Salmo trutta f. fario) to water downstream of a wastewater outfall (Schussen River) or to water from a reference site (Argen River) to investigate the vitellogenin production. Furthermore, two feral fish species, chub (Leuciscus cephalus) and spirlin (Alburnoides bipunctatus), were caught in both rivers to determine their gonadal maturity and the gonadosomatic index. Chemical analyses provided only little information about endocrine active substances, whereas the in vitro assays revealed endocrine potentials in most of the samples. In addition to endocrine potentials, we also observed toxic potentials (E-screen/reproduction test) in waste water samples, which could interfere with and camouflage endocrine effects. The results of our in vivo tests were mostly in line with the results of the in vitro assays and revealed a consistent reproduction-disrupting (reproduction tests) and an occasional endocrine action (vitellogenin levels) in both investigated rivers, with more pronounced effects for the Schussen river (e.g. a lower gonadosomatic index). We were able to show that biological in vitro assays for endocrine potentials in natural stream water reasonably reflect reproduction and endocrine disruption observed in snails and field-exposed fish, respectively. PMID:24901835

Space agriculture for habitation on Mars with hyper-thermophilic aerobic composting bacteria

NASA Astrophysics Data System (ADS)

Kanazawa, S.; Ishikawa, Y.; Tomita-Yokotani, K.; Hashimoto, H.; Kitaya, Y.; Yamashita, M.; Nagatomo, M.; Oshima, T.; Wada, H.; Space Agriculture Task Force, J.

Manned Mars exploration requires recycle of materials to support human life A conceptual design is developed for space agriculture which is driven by the biologically regenerative function Hyper-thermophilic aerobic composting bacterial ecology is the core of materials recycling system to process human metabolic waste and inedible biomass and convert them to fertilizer for plants cultivation A photosynthetic reaction of plants will be driven by solar energy Water will be recycled by cultivation of plants and passing it through plant bodies Sub-surface water and atmospheric carbon dioxide are the natural resource available on Mars and these resources will be converted to oxygen and foods We envision that the agricultural system will be scaled up by importing materials from Martian environment Excess oxygen will be obtained from growing trees for structural and other components Minor elements including N P K and other traces will be introduced as fertilizers or nutrients into the agricultural materials circulation Nitrogen will be collected from Martian atmosphere We will assess biological fixation of nitrogen using micro-organisms responsible in Earth biosphere Hyper-thermophilic aerobic bacterial ecology is effective to convert waste materials into useful forms to plants This microbial technology has been well established on ground for processing sewage and waste materials For instance the hyper-thermophilic bacterial system is applied to a composting machine in a size of a trash box in home kitchen Since such a home electronics

Derivation of the Korean radwaste scaling factor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kwang Yong Jee; Hong Joo Ahn; Se Chul Sohn

2007-07-01

The concentrations of several radionuclides in low and intermediate level radioactive waste (LILW) drums have to be determined before shipping to disposal facilities. A notice, by the Ministry of Science and Technology (MOST) of the Korean Government, related to the disposal of LILW drums came into effect at the beginning of 2005, with regards to a radionuclide regulation inside a waste drum. MOST allows for an indirect radionuclide assay using a scaling factor to measure the inventories due to the difficulty of nondestructively measuring the essential {alpha} and {beta}-emitting nuclides inside a drum. That is, a scaling factor calculated throughmore » a correlation of the {alpha} or {beta}-emitting nuclide (DTM, Difficult-To-Measure) with a {gamma}-emitting nuclide (ETM, Easy-To-Measure) which has systematically similar properties with DTM nuclides. In this study, radioactive wastes, such as spent resin and dry active waste which were generated at different sites of a PWR and a site of a PHWR type Korean NPP, were partially sampled and analyzed for regulated radionuclides by using radiochemical methods. According to a reactor type and a waste form, the analysis results of each radionuclide were classified. Korean radwaste scaling factor was derived from database of radionuclide concentrations. (authors)« less

Waste inspection tomography (WIT)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bernardi, R.T.

1996-12-31

WIT is a self-sufficient mobile semitrailer for nondestructive evaluation and nondestructive assay of nuclear waste drums using x-ray and gamma-ray tomography. The recently completed Phase I included the design, fabrication, and initial testing of all WIT subsystems installed on-board the trailer. Initial test results include 2 MeV digital radiography, computed tomography, Anger camera imaging, single photon emission computed tomography, gamma-ray spectroscopy, collimated gamma scanning, and active and passive computed tomography using a 1.4 mCi source of {sup 166}Ho. These techniques were initially demonstrated on a 55-gallon phantom drum with 3 simulated waste matrices of combustibles, heterogeneous metals, and cement usingmore » check sources of gamma active isotopes such as {sup 137}Cs and {sup 133}Ba with 9-250 {mu}Ci activities. Waste matrix identification, isotopic identification, and attenuation-corrected gamma activity determination were demonstrated nondestructively and noninvasively in Phase I. Currently ongoing Phase II involves DOE site field test demonstrations at LLNL, RFETS, and INEL with real nuclear waste drums. Current WIT experience includes 55 gallon drums of cement, graphite, sludge, glass, metals, and combustibles. Thus far WIT has inspected drums with 0-20 gms of {sup 239}Pu.« less

Rapid detection of porcine circovirus type 2 using a TaqMan-based real-time PCR.

PubMed

Zhao, Kai; Han, Fangting; Zou, Yong; Zhu, Lianlong; Li, Chunhua; Xu, Yan; Zhang, Chunling; Tan, Furong; Wang, Jinbin; Tao, Shiru; He, Xizhong; Zhou, Zongqing; Tang, Xueming

2010-12-31

Porcine circovirus type 2 (PCV2) and the associated disease postweaning multisystemic wasting syndrome (PMWS) have caused heavy losses in global agriculture in recent decades. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS. To establish a sensitive, specific assay for the detection and quantitation of PCV2, we designed and synthesized specific primers and a probe in the open reading frame 2. The assay had a wide dynamic range with excellent linearity and reliable reproducibility, and detected between 102 and 1010 copies of the genomic DNA per reaction. The coefficient of variation for Ct values varied from 0.59% to 1.05% in the same assay and from 1.9% to 4.2% in 10 different assays. The assay did not cross-react with porcine circovirus type 1, porcine reproductive and respiratory, porcine epidemic diarrhea, transmissible gastroenteritis of pigs and rotavirus. The limits of detection and quantitation were 10 and 100 copies, respectively. Using the established real-time PCR system, 39 of the 40 samples we tested were detected as positive.

Metabolic response of environmentally isolated microorganisms to industrial effluents: Use of a newly described cell culture assay

NASA Technical Reports Server (NTRS)

Ferebee, Robert N.

1992-01-01

An environmental application using a microtiter culture assay to measure the metabolic sensitivity of microorganisms to petrochemical effluents will be tested. The Biomedical Operations and Research Branch at NASA JSC has recently developed a rapid and nondestructive method to measure cell growth and metabolism. Using a colorimetric procedure the uniquely modified assay allows the metabolic kinetics of prokaryotic and eukaryotic cells to be measured. Use of such an assay if adapted for the routine monitoring of waste products, process effluents, and environmentally hazardous substances may prove to be invaluable to the industrial community. The microtiter method as described will be tested using microorganisms isolated from the Galveston Bay aquatic habitat. The microbial isolates will be identified prior to testing using the automated systems available at JSC. Sodium dodecyl sulfate (SDS), cadmium, and lead will provide control toxic chemicals. The toxicity of industrial effluent from two industrial sites will be tested. An effort will be made to test the efficacy of this assay for measuring toxicity in a mixed culture community.

Evaluation of radical scavenging activity, intestinal cell viability and antifungal activity of Brazilian propolis by-product.

PubMed

de Francisco, Lizziane; Pinto, Diana; Rosseto, Hélen; Toledo, Lucas; Santos, Rafaela; Tobaldini-Valério, Flávia; Svidzinski, Terezinha; Bruschi, Marcos; Sarmento, Bruno; Oliveira, M Beatriz P P; Rodrigues, Francisca

2018-03-01

Propolis is a natural adhesive resinous compound produced by honeybees to protect hives from bacteria and fungi, being extremely expensive for food industry. During propolis production, a resinous by-product is formed. This resinous waste is currently undervalued and underexploited. Accordingly, in this study the proximate physical and chemical quality, as well as the antioxidant activity, radical scavenging activity and cell viability of this by-product were evaluated and compared with propolis in order to boost new applications in food and pharmaceutical industries. The results revealed that the by-product meets the physical and chemical quality standards expected and showed that the propolis waste contains similar amounts of total phenolic content (TPC) and total flavonoid content (TFC) to propolis. Also, a good scavenging activity against reactive oxygen and nitrogen species (ROS and RNS, respectively) determined by the assays of superoxide anion radical (O 2 - ), hydrogen peroxide (H 2 O 2 ), hypochlorous acid (HOCl), nitric oxide (NO) and peroxyl radical (ROO) were determined. Linear positive correlations were established between the TPC of both samples and the antioxidant activity evaluated by three different methods (DPPH, ABTS and FRAP assays). The extracts were also screened for cell viability assays in two different intestinal cell lines (HT29-MTX and Caco-2), showing a viability concentration-dependent. Similarly, the Artemia salina assay, used to assess toxicity, demonstrated the concentration influence on results. Finally, the antifungal activity against ATCC species of Candida was demonstrated. These results suggest that propolis by-product can be used as a new rich source of bioactive compounds for different areas, such as food or pharmaceutical. Copyright © 2017 Elsevier Ltd. All rights reserved.

Automated Low-Cost Smartphone-Based Lateral Flow Saliva Test Reader for Drugs-of-Abuse Detection.

PubMed

Carrio, Adrian; Sampedro, Carlos; Sanchez-Lopez, Jose Luis; Pimienta, Miguel; Campoy, Pascual

2015-11-24

Lateral flow assay tests are nowadays becoming powerful, low-cost diagnostic tools. Obtaining a result is usually subject to visual interpretation of colored areas on the test by a human operator, introducing subjectivity and the possibility of errors in the extraction of the results. While automated test readers providing a result-consistent solution are widely available, they usually lack portability. In this paper, we present a smartphone-based automated reader for drug-of-abuse lateral flow assay tests, consisting of an inexpensive light box and a smartphone device. Test images captured with the smartphone camera are processed in the device using computer vision and machine learning techniques to perform automatic extraction of the results. A deep validation of the system has been carried out showing the high accuracy of the system. The proposed approach, applicable to any line-based or color-based lateral flow test in the market, effectively reduces the manufacturing costs of the reader and makes it portable and massively available while providing accurate, reliable results.

A Combined Fabrication and Instrumentation Platform for Sample Preparation.

PubMed

Guckenberger, David J; Thomas, Peter C; Rothbauer, Jacob; LaVanway, Alex J; Anderson, Meghan; Gilson, Dan; Fawcett, Kevin; Berto, Tristan; Barrett, Kevin; Beebe, David J; Berry, Scott M

2014-06-01

While potentially powerful, access to molecular diagnostics is substantially limited in the developing world. Here we present an approach to reduced cost molecular diagnostic instrumentation that has the potential to empower developing world communities by reducing costs through streamlining the sample preparation process. In addition, this instrument is capable of producing its own consumable devices on demand, reducing reliance on assay suppliers. Furthermore, this instrument is designed with an "open" architecture, allowing users to visually observe the assay process and make modifications as necessary (as opposed to traditional "black box" systems). This open environment enables integration of microfluidic fabrication and viral RNA purification onto an easy-to-use modular system via the use of interchangeable trays. Here we employ this system to develop a protocol to fabricate microfluidic devices and then use these devices to isolate viral RNA from serum for the measurement of human immunodeficiency virus (HIV) viral load. Results obtained from this method show significantly reduced error compared with similar nonautomated sample preparation processes. © 2014 Society for Laboratory Automation and Screening.

Effect of moisture of municipal biowaste on start-up and efficiency of mesophilic and thermophilic dry anaerobic digestion.

PubMed

Li, Chaoran; Mörtelmaier, Christoph; Winter, Josef; Gallert, Claudia

2014-09-01

Methane production from biowaste with 20-30% dry matter (DM) by box-type dry anaerobic digestion and contributing bacteria were determined for incubation at 20, 37 and 55 °C. The same digestion efficiency as for wet anaerobic digestion of biowaste was obtained for dry anaerobic digestion with 20% DM content at 20, 37 and 55 °C and with 25% DM content at 37 and 55 °C. No or only little methane was produced in dry anaerobic reactors with 30% DM at 20, 37 or 55 °C. Population densities in the 20-30% DM-containing biowaste reactors were similar although in mesophilic and thermophilic biowaste reactors with 30% DM content significantly less but phylogenetically more diverse archaea existed. Biogas production in the 20% and 25% DM assays was catalyzed by Methanosarcinales and Methanomicrobiales. In all assays Pelotomaculum and Syntrophobacter species were dominant propionate degraders. Copyright © 2014 Elsevier Ltd. All rights reserved.

MdHY5 positively regulates cold tolerance via CBF-dependent and CBF-independent pathways in apple.

PubMed

An, Jian-Ping; Yao, Ji-Fang; Wang, Xiao-Na; You, Chun-Xiang; Wang, Xiao-Fei; Hao, Yu-Jin

2017-11-01

Cold stress is a major external stimulator that affects crop quality and productivity. The CBF cold regulatory pathway has been regarded as a master regulator in the response to cold stress. In this study, we found that the apple bZIP transcription factor, MdHY5, was responsive to cold treatment both at the transcriptional and at the post-translational levels. Moreover, overexpression of MdHY5 enhanced cold tolerance in apple calli and Arabidopsis. Subsequently, EMSA assay and transient expression assay demonstrated that MdHY5 positively regulated the transcript of MdCBF1 by binding to G-Box motif of its promoter. Furthermore, MdHY5 also regulated the expression of CBF-independent cold-regulated genes. Taken together, our data suggest that MdHY5 positively modulates plant cold tolerance through CBF-dependent and CBF-independent pathways, providing a deeper understanding of MdHY5-regulated cold tolerance in apple. Copyright © 2017 Elsevier GmbH. All rights reserved.

Seafood-Processing Sludge Composting: Changes to Microbial Communities and Physico-Chemical Parameters of Static Treatment versus for Turning during the Maturation Stage

PubMed Central

Alves, David; Mato, Salustiano

2016-01-01

In general, in composting facilities the active, or intensive, stage of the process is done separately from the maturation stage, using a specific technology and time. The pre-composted material to be matured can contain enough biodegradable substrates to cause microbial proliferation, which in turn can cause temperatures to increase. Therefore, not controlling the maturation period during waste management at an industrial level can result in undesired outcomes. The main hypothesis of this study is that controlling the maturation stage through turning provides one with an optimized process when compared to the static approach. The waste used was sludge from a seafood-processing plant, mixed with shredded wood (1:2, v/v). The composting system consists of an intensive stage in a 600L static reactor, followed by maturation in triplicate in 200L boxes for 112 days. Two tests were carried out with the same process in reactor and different treatments in boxes: static maturation and turning during maturation when the temperature went above 55°C. PLFAs, organic matter, pH, electrical conductivity, forms of nitrogen and carbon, hydrolytic enzymes and respiratory activity were periodically measured. Turning significantly increased the duration of the thermophilic phase and consequently increased the organic-matter degradation. PCA differentiated significantly the two treatments in function of tracking parameters, especially pH, total carbon, forms of nitrogen and C/N ratio. So, stability and maturity optimum values for compost were achieved in less time with turnings. Whereas turning resulted in microbial-group stabilization and a low mono/sat ratio, static treatment produced greater variability in microbial groups and a high mono/sat ratio, the presence of more degradable substrates causes changes in microbial communities and their study during maturation gives an approach of the state of organic-matter degradation. Obtaining quality compost and optimizing the composting process requires using turning as a control mechanism during maturation. PMID:28002444

Preliminary Results from Electric Arc Furnace Off-Gas Enthalpy Modeling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nimbalkar, Sachin U; Thekdi, Arvind; Keiser, James R

2015-01-01

This article describes electric arc furnace (EAF) off-gas enthalpy models developed at Oak Ridge National Laboratory (ORNL) to calculate overall heat availability (sensible and chemical enthalpy) and recoverable heat values (steam or power generation potential) for existing EAF operations and to test ORNL s new EAF waste heat recovery (WHR) concepts. ORNL s new EAF WHR concepts are: Regenerative Drop-out Box System and Fluidized Bed System. The two EAF off-gas enthalpy models described in this paper are: 1.Overall Waste Heat Recovery Model that calculates total heat availability in off-gases of existing EAF operations 2.Regenerative Drop-out Box System Model in whichmore » hot EAF off-gases alternately pass through one of two refractory heat sinks that store heat and then transfer it to another gaseous medium These models calculate the sensible and chemical enthalpy of EAF off-gases based on the off-gas chemical composition, temperature, and mass flow rate during tap to tap time, and variations in those parameters in terms of actual values over time. The models provide heat transfer analysis for the aforementioned concepts to confirm the overall system and major component sizing (preliminary) to assess the practicality of the systems. Real-time EAF off-gas composition (e.g., CO, CO2, H2, and H2O), volume flow, and temperature data from one EAF operation was used to test the validity and accuracy of the modeling work. The EAF off-gas data was used to calculate the sensible and chemical enthalpy of the EAF off-gases to generate steam and power. The article provides detailed results from the modeling work that are important to the success of ORNL s EAF WHR project. The EAF WHR project aims to develop and test new concepts and materials that allow cost-effective recovery of sensible and chemical heat from high-temperature gases discharged from EAFs.« less

Gamma neutron assay method and apparatus

DOEpatents

Cole, J.D.; Aryaeinejad, R.; Greenwood, R.C.

1995-01-03

The gamma neutron assay technique is an alternative method to standard safeguards techniques for the identification and assaying of special nuclear materials in a field or laboratory environment, as a tool for dismantlement and destruction of nuclear weapons, and to determine the isotopic ratios for a blend-down program on uranium. It is capable of determining the isotopic ratios of fissionable material from the spontaneous or induced fission of a sample to within approximately 0.5%. This is based upon the prompt coincidence relationships that occur in the fission process and the proton conservation and quasi-conservation of nuclear mass (A) that exists between the two fission fragments. The system is used in both passive (without an external neutron source) and active (with an external neutron source) mode. The apparatus consists of an array of neutron and gamma-ray detectors electronically connected to determine coincident events. The method can also be used to assay radioactive waste which contains fissile material, even in the presence of a high background radiation field. 7 figures.

Gamma neutron assay method and apparatus

DOEpatents

Cole, Jerald D.; Aryaeinejad, Rahmat; Greenwood, Reginald C.

1995-01-01

The gamma neutron assay technique is an alternative method to standard safeguards techniques for the identification and assaying of special nuclear materials in a field or laboratory environment, as a tool for dismantlement and destruction of nuclear weapons, and to determine the isotopic ratios for a blend-down program on uranium. It is capable of determining the isotopic ratios of fissionable material from the spontaneous or induced fission of a sample to within approximately 0.5%. This is based upon the prompt coincidence relationships that occur in the fission process and the proton conservation and quasi-conservation of nuclear mass (A) that exists between the two fission fragments. The system is used in both passive (without an external neutron source and active (with an external neutron source) mode. The apparatus consists of an array of neutron and gamma-ray detectors electronically connected to determine coincident events. The method can also be used to assay radioactive waste which contains fissile material, even in the presence of a high background radiation field.

CSER-98-002: Criticality analysis for the storage of special nuclear material sources and standards in the WRAP facility

DOE Office of Scientific and Technical Information (OSTI.GOV)

GOLDBERG, H.J.

1999-05-18

The Waste Receiving and Processing (WRAP) Facility will store uranium and transuranic (TRU) sources and standards for certification that WRAP meets the requirements of the Quality Assurance Program Plan (QAPP) for the Waste Isolation Pilot Plant (WIPP). In addition, WRAP must meet internal requirements for testing and validation of measuring instruments for nondestructive assay (NDA). In order to be certified for WIPP, WRAP will participate in the NDA Performance Demonstration Program (PDP). This program is a blind test of the NDA capabilities for TRU waste. It is intended to ensure that the NDA capabilities of this facility satisfy the requirementsmore » of the quality assurance program plan for the WIPP. The PDP standards have been provided by the Los Alamos National Laboratory (LANL) for this program. These standards will be used in the WRAP facility.« less

Enhancing biogas production from anaerobic biodegradation of the organic fraction of municipal solid waste through leachate blending and recirculation.

PubMed

Nair, Arjun; Sartaj, Majid; Kennedy, Kevin; Coelho, Nuno M G

2014-10-01

Leachate recirculation has a profound advantage on biodegradation of the organic fraction of municipal solid waste in landfills. Mature leachate from older sections of landfills (>10 years) and young leachate were blended and added to organic fraction of municipal solid waste in a series of biomethane potential assay experiments with different mixing ratios of mature and young leachate and their effect on biogas production was monitored. The improvement in biogas production was in the range of 19%-41% depending on the ratio of mixing old and new leachate. The results are conclusive that the biogas generation could be improved by blending the old and new leachate in a bioreactor landfill system as compared with a conventional system employed in bioreactor landfills today for recirculating the same age leachate. © The Author(s) 2014.

Shikonins, phytocompounds from Lithospermum erythrorhizon, inhibit the transcriptional activation of human tumor necrosis factor alpha promoter in vivo.

PubMed

Staniforth, Vanisree; Wang, Sheng-Yang; Shyur, Lie-Fen; Yang, Ning-Sun

2004-02-13

Tumor necrosis factor alpha (TNF-alpha) contributes to the pathogenesis of both acute and chronic inflammatory diseases and has been a target for the development of new anti-inflammatory drugs. Shikonins, the naphthoquinone pigments present in the root tissues of Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae), have been reported to exert anti-inflammatory effects both in vitro and in vivo. In this study, we evaluated the effects of shikonin and its derivatives on the transcriptional activation of human TNF-alpha promoter in a gene gun-transfected mouse skin system by using a luciferase reporter gene assay. The crude plant extract of L. erythrorhizon as well as derived individual compounds shikonin, isobutyryl shikonin, acetyl shikonin, dimethylacryl shikonin and isovaleryl shikonin showed significant dose-dependent inhibition of TNF-alpha promoter activation. Among the tested compounds, shikonin and isobutyryl shikonin exhibited the highest inhibition of TNF-alpha promoter activation and also showed significant suppression of transgenic human TNF-alpha mRNA expression and protein production. We demonstrated that shikonin-inhibitory response was retained in the core TNF-alpha promoter region containing the TATA box and a 48-bp downstream sequence relative to the transcription start site. Further our results indicated that shikonin suppressed the basal transcription and activator-regulated transcription of TNF-alpha by inhibiting the binding of transcription factor IID protein complex (TATA box-binding protein) to TATA box. These in vivo results suggest that shikonins inhibit the transcriptional activation of the human TNF-alpha promoter through interference with the basal transcription machinery. Thus, shikonins may have clinical potential as anti-inflammatory therapeutics.

Genomic Organization, Phylogenetic and Expression Analysis of the B-BOX Gene Family in Tomato

PubMed Central

Chu, Zhuannan; Wang, Xin; Li, Ying; Yu, Huiyang; Li, Jinhua; Lu, Yongen; Li, Hanxia; Ouyang, Bo

2016-01-01

The B-BOX (BBX) proteins encode a class of zinc-finger transcription factors possessing one or two B-BOX domains and in some cases an additional CCT (CO, CO-like and TOC1) motif, which play important roles in regulating plant growth, development and stress response. Nevertheless, no systematic study of BBX genes has undertaken in tomato (Solanum lycopersicum). Here we present the results of a genome-wide analysis of the 29 BBX genes in this important vegetable species. Their structures, conserved domains, phylogenetic relationships, subcellular localizations, and promoter cis-regulatory elements were analyzed; their tissue expression profiles and expression patterns under various hormones and stress treatments were also investigated in detail. Tomato BBX genes can be divided into five subfamilies, and twelve of them were found to be segmentally duplicated. Real-time quantitative PCR analysis showed that most BBX genes exhibited different temporal and spatial expression patterns. The expression of most BBX genes can be induced by drought, polyethylene glycol-6000 or heat stress. Some BBX genes were induced strongly by phytohormones such as abscisic acid, gibberellic acid, or ethephon. The majority of tomato BBX proteins was predicted to be located in nuclei, and the transient expression assay using Arabidopsis mesophyll protoplasts demonstrated that all the seven BBX members tested (SlBBX5, 7, 15, 17, 20, 22, and 24) were localized in nucleus. Our analysis of tomato BBX genes on the genome scale would provide valuable information for future functional characterization of specific genes in this family. PMID:27807440

Promoter/repressor system of Lactobacillus plantarum phage og1e: characterization of the promoters pR49-pR-pL and overproduction of the cro-like protein cng in Escherichia coli.

PubMed

Kakikawa, M; Watanabe, N; Funawatashi, T; Oki, M; Yasukawa, H; Taketo, A; Kodaira, K I

1998-07-30

The Lactobacillus plantarum phage og1e (42259bp) has two repressor-like genes cng and cpg oriented oppositely, accompanied by three potential promoters pR, pL and pR49, and seven operator-like sequences (GATAC-boxes) (Kodaira et al., 1997). In this study, the og1e putative promoters were introduced into the Escherichia coli promoter-detecting plasmid pKK232-8. In E. coli CK111, pR (pKPR1), pL (pKPL1) and pR49 (pKPR49) exhibited distinct CAT activities. When pKPR1 or pKPL1 was coexistent with a compatible plasmid pACYC184 carrying pR-cng (pA4PRCN1), the CAT activity was decreased significantly. On the other hand, cng directed a protein (Cng) of 10.1 kDa in E. coli under the control of T7 promoter. Gel mobility-shift assays demonstrated that Cng binds specifically to a DNA region containing the GATAC-boxes. In addition, primer extension analyses demonstrated that the two sequences pR and pL act as a promoter in L. plantarum as well as in E. coli. These results suggested that the potential promoters pR and pL probably function for the lytic and lysogenic pathways, respectively, and Cng may act as a repressor presumably through the GATAC-boxes as operators.

Angiogenesis in tissue engineering: from concept to the vascularization of scaffold construct

NASA Astrophysics Data System (ADS)

Amirah Ishak, Siti; Pangestu Djuansjah, J. R.; Kadir, M. R. Abdul; Sukmana, Irza

2014-06-01

Angiogenesis, the formation of micro-vascular network from the preexisting vascular vessels, has been studied in the connection to the normal developmental process as well as numerous diseases. In tissue engineering research, angiogenesis is also essential to promote micro-vascular network inside engineered tissue constructs, mimicking a functional blood vessel in vivo. Micro-vascular network can be used to maintain adequate tissue oxygenation, nutrient transfer and waste removal. One of the problems faced by angiogenesis researchers is to find suitable in vitro assays and methods for assessing the effect of regulators on angiogenesis and micro-vessel formation. The assay would be reliable and repeatable with easily quantifiable with physiologically relevant. This review aims to highlights recent advanced and future challenges in developing and using an in vitro angiogenesis assay for the application on biomedical and tissue engineering research.

Phenotypic and genotypic characterization of clinically relevant bacteria isolated from dental waste and waste workers' hands, mucosas and coats.

PubMed

Tagliaferri, T L; Vieira, C D; de Carvalho, M A R; Ladeira, L C D; Magalhães, P P; de Macêdo Farias, L; Dos Santos, S G

2017-10-01

Infectious wastes are potential sources of pathogenic micro-organisms, which may represent a risk to the professionals who manage them. In this study, we aimed to characterize the infectious bacteria present in dental waste and waste workers. The dental waste produced over 24 h was collected and waste workers were sampled by swabbing. Isolate resistance profiles were characterized by Vitek ® and PCR and biofilm formation by Congo Red agar, string test and microtitre assay. To assess similarity between the waste and the workers' samples, a random amplified polymorphic DNA test was used. Twenty-eight bacteria were identified as clinically relevant. The most frequent gene was bla TEM present in five Gram-negative micro-organisms, and one bla SHV in Klebsiella pneumoniae. All Pseudomonas aeruginosa were positive to extracellular polymeric substances formation, except one isolated from a worker. Klebsiella pneumoniae had negative results for the string test. Pseudomonas aeruginosa showed better adherence at 25°C after 48 h of incubation and K. pneumonia had the best biofilm formation at the same temperature, after 24 h. The similarity between P. aeruginosa recovered from dental waste and from workers was low, however, it is important to note that a pathogen was found on a worker's hands and that improvements in biosafety are required. Infectious dental waste can contain clinically relevant bacteria with important resistance and biofilm profiles. These micro-organisms could be transmitted to waste workers, other professionals and patients if the principles of biosafety measures are neglected. To our knowledge, no study has ever evaluated the microbial characterization and the potential contamination risk of dental infectious waste and waste handlers. The presence of clinically relevant bacteria in the hands and nasal mucosa of waste workers highlights the need for studies in this field to clarify the risk of these pathogens in dental healthcare services, and to stress the need for an efficient waste management. © 2017 The Society for Applied Microbiology.

Development and evaluation of a culture-independent method for source determination of fecal wastes in surface and storm waters using reverse transcriptase-PCR detection of FRNA coliphage genogroup gene sequences.

EPA Science Inventory

A complete method, incorporating recently improved reverse transcriptase-PCR primer/probe assays and including controls for determining interferences to phage recoveries from water sample concentrates and for detecting interferences to their analysis, was developed for the direct...

Design and Assessment of a Real Time Reverse Transcription-PCR Method to Genotype Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae).

EPA Science Inventory

A real-time, reverse transcription-PCR (RT-qPCR) assay was developed to differentiate the four genogroups of male-specific ssRNA coliphages (FRNA) (family Leviviridae). As FRNA display a trend of source-specificity (human sewage or animal waste) at the genogroup level, this assa...

Development and evaluation of a culture-independent method for source determination of fecal wastes in surface and storm waters using reverse transcriptase-PCR detection of FRNA coliphage genogroup gene sequences

EPA Science Inventory

A complete method, incorporating recently improved reverse transcriptase-PCR primer/probe assays and including controls for determining interferences to phage recoveries from water sample concentrates and for detecting interferences to their analysis, was developed for the direct...

Anti and Androgenic Activities in MDA-KB2 Cells: A Comparison of Performance in 96 Well Versus HTS Assays

EPA Science Inventory

We developed the MDA-kb2 cell line to screen androgen agonists/antagonists (Wilson et al., ToxSci 66:69, 2002). MDA-kb2 has been used to quantify anti- and androgenic activities of chemicals, mixtures, combustion by-products, oil dispersants and waste, source and drinking water s...

Comparison of an Assay for Dehalococcoides DNA and a Microcosm Study in Predicting Reductive Dechlorination of Chlorinated Ethenes in the field

EPA Science Inventory

The study aims to compare the detection of 16S rRNA gene of Dehalococcoides species and the microcosm study for biotransformation in predicting reductive dechlorination of chlorinated ethylenes in ground water at hazardous waste sites. A total of 72 ground water samples were coll...

Transuranic solid waste management programs. Progress report, July--December 1975

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1976-09-01

Progress is reported for three transuranic solid waste management programs funded at the Los Alamos Scientific Laboratory (LASL) by the Energy Research and Development Administration (ERDA) Division of Fuel Cycle and Production (NFCP). Under the Transuranic Waste Research and Development Program, continued studies have shown the potential attractiveness of fiber drums as an acceptable substitute for the current mild steel storage containers. Various fire retardants have been evaluated, with one indicating significant ability to inhibit fire propagation. Continued radiolysis studies, under laboratory and field conditions, continue to reaffirm earlier LASL results indicating no significant hazard from radiolytic reactions, assuming nomore » change in current allowable loadings. Care must be exercised to differentiate between radiolytic and chemical reactions. Other efforts have identified a modification of chemical processing to reduce the amounts of plutonium requiring retrievable storage. Studies are also in progress to enhance the sensitivity of the LASL MEGAS assay system. The Transuranic-Contaminated Solid Waste Treatment Development Facility building was 72 percent complete as of December 31, 1975, which is in accord with the existing schedule. Procurement of process components is also on schedule. Certain modifications to the facility have been made, and various pre-facility experiments on waste container handling and processing have been completed. The program for the Evaluation of Transuranic-Contaminated Radioactive Waste Disposal Areas continued development of various computer modules for simulation of radionuclide transport within the biosphere. In addition, program staff contributed to an ERDA document on radioactive waste management through the preparation of a report on burial of radioactive waste at ERDA-contractor and commercial sites.« less

Co-digestion of livestock effluents, energy crops and agro-waste: feeding and process optimization in mesophilic and thermophilic conditions.

PubMed

Giuliano, A; Bolzonella, D; Pavan, P; Cavinato, C; Cecchi, F

2013-01-01

In this study the optimization of the biogas yield from anaerobic co-digestion of manures and energy crops was carried out using four pilot scale CSTRs under different operating conditions. The effect on biogas yield of the partial substitution of energy crops with agro-waste was also investigated. For each substrate used during the continuous trials, BMP batch assays were also carried out to verify the maximum methane yield theoretically obtainable. Continuous operation results indicated that the co-digestion of manures, energy crops and agro-waste was viable at all operating conditions tested, with the greatest specific gas production of 0.54 m(3)/kg VS(fed) at an organic load rate of 2 kg TVS/m(3)(r)d consisting of 50% manure, 25% energy crops and 25% agro-waste on VS basis. No significant differences were observed between high and low loaded reactors suggesting the possibility of either improving the OLR in existing anaerobic reactors or reducing the design volumes of new reactors. Copyright © 2012 Elsevier Ltd. All rights reserved.

Chromosomal aberrations and DNA damage in human populations exposed to the processing of electronics waste.

PubMed

Liu, Qiang; Cao, Jia; Li, Ke Qiu; Miao, Xu Hong; Li, Guang; Fan, Fei Yue; Zhao, Yong Cheng

2009-05-01

It has been known that the pollutants of electronic wastes (E-wastes) can lead to severe pollution to the environment. It has been reported that about 50% to 80% of E-wastes from developed countries are exported to Asia and Africa. It has become a major global environmental problem to deal with 'E-wastes'. E-waste recycling has remained primitive in Jinghai, China. This not only produces enormous environmental pollution but also can bring about toxic or genotoxic effects on the human body, threatening the health of both current residents and future generations living in the local environment. The concentration of lead in the blood of children in the E-waste polluted area in China is higher than that of the control area. But little is known about the cytogenetic effect to human beings caused by the pollution of E-wastes. In the present study, experiments have been performed to investigate the genetics of permanent residents of three villages with numerous E-waste disposal sites and to analyze the harmful effects of exposure to E-wastes. In total, 171 villagers (exposed group) were randomly selected from permanent residents of three villages located in Jinghai County of Tianjin, China, where there has been massive disposal of E-wastes. Thirty villagers were selected from the neighboring towns without E-waste disposal sites to serve as controls. Chromosomal aberrations and cytokinesis blocking micronucleus were performed to detect the cytogenetic effect, dic + r (dicentric and ring chromosome), monomer, fragments (acentric fragments, minute chromosomes, and acentric rings), translocation, satellite, quadriradial, total aberrations, and micronuclear rate were scored for each subject. DNA damage was detected using comet assay; the DNA percentage in the comet tail (TDNA%), tail moment (TM), and Olive tail moment (OTM) were recorded to describe DNA damage to lymphocytes. The total chromosome aberration rates (5.50%) and micronuclear rates (16.99%) of the exposure group were significantly higher than in the control group (P = 0.000). The percentage of DNA in the comet tail, tail moment, and Olive tail moment detected by comet assay showed that there was a significant difference in DNA damage in the exposure group (P = 0.000). The chromosome aberration, micronucleus rate, and DNA damage observed in women were significantly higher than those in men. Chromosome aberration and micronuclear rates of both smokers and non-smokers in the exposure group are obviously higher than that in the control group (P = 0.000). The use of outdated (and unsafe) ways to deal with E-wastes can lead to exposure to a variety of substances harmful to human health. The components of pollution may enter the human body through the air, drinking water, and food chain to damage human genetic material, resulting in genomic instability. The rates of chromosomal aberration, micronucleus formation, and the degree of DNA damage in women in the group exposed to electronic waste were significantly higher than in men. The reason for this may be concerned with the traditional lifestyle of the local residents or the difference of sensitivity to the exposure to E-wastes or any others. Further investigations are needed to provide evidence to demonstrate this. Here, we report the obviously cytogenetic toxicity to the exposure population by the E-waste pollution for the first time. E-waste pollution may be a potential agent of genetic mutation, and may induce cytogenetic damage within the general population exposed to the pollution. These findings need to be considered, and steps should be taken to protect the current population and future generations from the effects of pollution with E-wastes. The above results remind us that the impact of E-waste recycling on environmental quality of Jinghai should be evaluated soon. Moreover, it is urgent for the government to prohibit E-waste import and its processing by outdated ways. The future studies such as pollutant details of drinking water, air, and soil in the area as well as epidemiological investigations on the harmful effect to children must be performed eagerly. All the data available do provide a compelling case for immediate action in both countries to address workplace health and safety and waste management.

SRY-box-containing Gene 2 Regulation of Nuclear Receptor Tailless (Tlx) Transcription in Adult Neural Stem Cells*

PubMed Central

Shimozaki, Koji; Zhang, Chun-Li; Suh, Hoonkyo; Denli, Ahmet M.; Evans, Ronald M.; Gage, Fred H.

2012-01-01

Adult neurogenesis is maintained by self-renewable neural stem cells (NSCs). Their activity is regulated by multiple signaling pathways and key transcription factors. However, it has been unclear whether these factors interplay with each other at the molecular level. Here we show that SRY-box-containing gene 2 (Sox2) and nuclear receptor tailless (TLX) form a molecular network in adult NSCs. We observed that both Sox2 and TLX proteins bind to the upstream region of Tlx gene. Sox2 positively regulates Tlx expression, whereas the binding of TLX to its own promoter suppresses its transcriptional activity in luciferase reporter assays. Such TLX-mediated suppression can be antagonized by overexpressing wild-type Sox2 but not a mutant lacking the transcriptional activation domain. Furthermore, through regions involved in DNA-binding activity, Sox2 and TLX physically interact to form a complex on DNAs that contain a consensus binding site for TLX. Finally, depletion of Sox2 revealed the potential negative feedback loop of TLX expression that is antagonized by Sox2 in adult NSCs. These data suggest that Sox2 plays an important role in Tlx transcription in cultured adult NSCs. PMID:22194602

SRY-box-containing gene 2 regulation of nuclear receptor tailless (Tlx) transcription in adult neural stem cells.

PubMed

Shimozaki, Koji; Zhang, Chun-Li; Suh, Hoonkyo; Denli, Ahmet M; Evans, Ronald M; Gage, Fred H

2012-02-17

Adult neurogenesis is maintained by self-renewable neural stem cells (NSCs). Their activity is regulated by multiple signaling pathways and key transcription factors. However, it has been unclear whether these factors interplay with each other at the molecular level. Here we show that SRY-box-containing gene 2 (Sox2) and nuclear receptor tailless (TLX) form a molecular network in adult NSCs. We observed that both Sox2 and TLX proteins bind to the upstream region of Tlx gene. Sox2 positively regulates Tlx expression, whereas the binding of TLX to its own promoter suppresses its transcriptional activity in luciferase reporter assays. Such TLX-mediated suppression can be antagonized by overexpressing wild-type Sox2 but not a mutant lacking the transcriptional activation domain. Furthermore, through regions involved in DNA-binding activity, Sox2 and TLX physically interact to form a complex on DNAs that contain a consensus binding site for TLX. Finally, depletion of Sox2 revealed the potential negative feedback loop of TLX expression that is antagonized by Sox2 in adult NSCs. These data suggest that Sox2 plays an important role in Tlx transcription in cultured adult NSCs.

Isolation and characterization of StERF transcription factor genes from potato (Solanum tuberosum L.).

PubMed

Wang, Zemin; Zhang, Ning; Zhou, Xiangyan; Fan, Qiang; Si, Huaijun; Wang, Di

2015-04-01

Ethylene response factor (ERF) is a major subfamily of the AP2/ERF family and plays significant roles in the regulation of abiotic- and biotic-stress responses. ERF proteins can interact with the GCC-box cis-element and then initiate a transcriptional cascade activating downstream ethylene response and enhancing plant stress tolerance. In this research, we cloned five StERF genes from potato (Solanum tuberosum L.). The expressional analysis of StERF genes revealed that they showed tissue- or organ-specific expression patterns and the expression levels in leaf, stem, root, flower, and tuber were different. The assays of quantitative real-time polymerase chain reaction (qRT-PCR) and the reverse transcription-PCR (RT-PCR) showed that the expression of five StERF genes was regulated by ethephon, methyl jasmonate (MeJA), salt and drought stress. The result from the yeast one-hybrid experiment showed that five StERFs had trans-activation activity and could specifically bind to the GCC-box cis-elements. The StERFs responded to abiotic factors and hormones suggested that they possibly had diverse roles in stress and hormone regulation of potato. Copyright © 2015 Académie des sciences. Published by Elsevier SAS. All rights reserved.

Cellular DEAD-box RNA helicase DDX6 modulates interaction of miR-122 with the 5' untranslated region of hepatitis C virus RNA.

PubMed

Biegel, Jason M; Henderson, Eric; Cox, Erica M; Bonenfant, Gaston; Netzband, Rachel; Kahn, Samantha; Eager, Rachel; Pager, Cara T

2017-07-01

Hepatitis C virus (HCV) subverts the cellular DEAD-box RNA helicase DDX6 to promote virus infection. Using polysome gradient analysis and the subgenomic HCV Renilla reporter replicon genome, we determined that DDX6 does not affect HCV translation. Rather expression of the subgenomic HCV Renilla luciferase reporter at late times, as well as labeling of newly synthesized viral RNA with 4-thiouridine showed that DDX6 modulates replication. Because DDX6 is an effector protein of the microRNA pathway, we also investigated its role in miR-122-directed HCV gene expression. Similar to sequestering miR-122, depletion of DDX6 modulated HCV RNA stability. Interestingly, miR-122-HCV RNA interaction assays with mutant HCV genomes sites and compensatory exogenous miR-122 showed that DDX6 affects the function of miR-122 at one particular binding site. We propose that DDX6 facilitates the miR-122 interaction with HCV 5' UTR, which is necessary for stabilizing the viral genome and the switch between translation and replication. Copyright © 2017 Elsevier Inc. All rights reserved.

Identification of a novel box C/D snoRNA from mouse nucleolar cDNA library.

PubMed

Zhou, Hui; Zhao, Jin; Yu, Chuan-He; Luo, Qing-Jun; Chen, Yue-Qin; Xiao, Yu; Qu, Liang-Hu

2004-02-18

By construction and screen of mouse nucleolar cDNA library, a novel mammalian small nucleolar RNAs (snoRNA) was identified. The novel snoRNA, 70 nt in length, displays structural features typical of C/D box snoRNA family. The snoRNA possesses an 11-nt-long rRNA antisense element and is predicted to guide the 2'-O-methylation of mouse 28S rRNA at G4043, a site unknown so far to be modified in vertebrates. The comparison of functional element of snoRNA guides among eukaryotes reveals that the novel snoRNA is a mammalian counterpart of yeast snR38 despite highly divergent sequence between them. Mouse and human snR38 and other cognates in distant vertebrates were positively detected with slight length variability. As expected, the rRNA ribose-methylation site predicted by mouse snR38 was precisely mapped by specific-primer extension assay. Furthermore, our analyses show that mouse and human snR38 gene have multiple variants and are nested in the introns of different host genes with unknown function. Thus, snR38 is a phylogenetically conserved methylation guide but exhibits different genomic organization in eukaryotes.

Identification of the domains in cyclin A required for binding to, and activation of, p34cdc2 and p32cdk2 protein kinase subunits.

PubMed Central

Kobayashi, H; Stewart, E; Poon, R; Adamczewski, J P; Gannon, J; Hunt, T

1992-01-01

The binding of cyclin A to p34cdc2 and p32cdk2 and the protein kinase activity of the complexes has been measured by cell-free translation of the corresponding mRNA in extracts of frog eggs, followed by immunoprecipitation. A variety of mutant cyclin A molecules have been constructed and tested in this assay. Small deletions and point mutations of highly conserved residues in the 100-residue "cyclin box" abolish binding and activation of both p34cdc2 and p32cdk2. By contrast, large deletions at the N-terminus have no effect on kinase binding and activation, until they remove residues beyond 161, where the first conserved amino acids are found in all known examples of cyclin A. At the C-terminus, removal of 14 or more amino acids abolishes activity. We also demonstrate that deletion of, or point mutations, in the cyclin A homologue of the 10-residue "destruction box," previously described in cyclin B (Glotzer et al., 1991) abolish cyclin proteolysis at the transition from M-phase to interphase. Images PMID:1333843

Identification of the domains in cyclin A required for binding to, and activation of, p34cdc2 and p32cdk2 protein kinase subunits.

PubMed

Kobayashi, H; Stewart, E; Poon, R; Adamczewski, J P; Gannon, J; Hunt, T

1992-11-01

The binding of cyclin A to p34cdc2 and p32cdk2 and the protein kinase activity of the complexes has been measured by cell-free translation of the corresponding mRNA in extracts of frog eggs, followed by immunoprecipitation. A variety of mutant cyclin A molecules have been constructed and tested in this assay. Small deletions and point mutations of highly conserved residues in the 100-residue "cyclin box" abolish binding and activation of both p34cdc2 and p32cdk2. By contrast, large deletions at the N-terminus have no effect on kinase binding and activation, until they remove residues beyond 161, where the first conserved amino acids are found in all known examples of cyclin A. At the C-terminus, removal of 14 or more amino acids abolishes activity. We also demonstrate that deletion of, or point mutations, in the cyclin A homologue of the 10-residue "destruction box," previously described in cyclin B (Glotzer et al., 1991) abolish cyclin proteolysis at the transition from M-phase to interphase.

MiRNA-101 inhibits oral squamous-cell carcinoma growth and metastasis by targeting zinc finger E-box binding homeobox 1

PubMed Central

Wu, Baolei; Lei, Delin; Wang, Lei; Yang, Xinjie; Jia, Sen; Yang, Zihui; Shan, Chun; Yang, Xi; Zhang, Chenping; Lu, Bin

2016-01-01

MicroRNAs (miRNAs) are implicated in the pathogenesis of oral squamous-cell carcinoma (OSCC). miR-101 is involved in the development and progression of OSCC, but the biological functions and underlying molecular mechanisms of this miRNA remain largely unknown. In this study, we showed that miR-101 was underexpressed in OSCC tissues and cell lines. miR-101 downregulation was inversely correlated with zinc finger E-box binding homeobox 1 (ZEB1) expression, lymph-node metastasis, and poor prognosis in OSCC patients. Enhanced expression of miR-101 significantly inhibited OSCC cell proliferation, apoptosis resistance, migration and invasion in vitro, and suppressed tumor growth and lung metastasis in vivo. Bioinformatics analyses showed that miR-101 directly targeted ZEB1, as confirmed by a dual-luciferase reporter assay. The inhibitory effects of miR-101 on OSCC growth and metastasis were attenuated and phenocopied by ZEB1 overexpression and knockdown, respectively. Overall, our findings indicated that miRNA-101 reduced OSCC growth and metastasis by targeting ZEB1 and provided new evidence of miR-101 as a potential therapeutic target for OSCC patients. PMID:27429852

Bone Morphogenetic Protein-Induced Msx1 and Msx2 Inhibit Myocardin-Dependent Smooth Muscle Gene Transcription▿

PubMed Central

Hayashi, Ken'ichiro; Nakamura, Seiji; Nishida, Wataru; Sobue, Kenji

2006-01-01

During the onset and progression of atherosclerosis, the vascular smooth muscle cell (VSMC) phenotype changes from differentiated to dedifferentiated, and in some cases, this change is accompanied by osteogenic transition, resulting in vascular calcification. One characteristic of dedifferentiated VSMCs is the down-regulation of smooth muscle cell (SMC) marker gene expression. Bone morphogenetic proteins (BMPs), which are involved in the induction of osteogenic gene expression, are detected in calcified vasculature. In this study, we found that the BMP2-, BMP4-, and BMP6-induced expression of Msx transcription factors (Msx1 and Msx2) preceded the down-regulation of SMC marker expression in cultured differentiated VSMCs. Either Msx1 or Msx2 markedly reduced the myocardin-dependent promoter activities of SMC marker genes (SM22α and caldesmon). We further investigated interactions between Msx1 and myocardin/serum response factor (SRF)/CArG-box motif (cis element for SRF) using coimmunoprecipitation, gel-shift, and chromatin immunoprecipitation assays. Our results showed that Msx1 or Msx2 formed a ternary complex with SRF and myocardin and inhibited the binding of SRF or SRF/myocardin to the CArG-box motif, resulting in inhibition of their transcription. PMID:17030628

Bone morphogenetic protein-induced MSX1 and MSX2 inhibit myocardin-dependent smooth muscle gene transcription.

PubMed

Hayashi, Ken'ichiro; Nakamura, Seiji; Nishida, Wataru; Sobue, Kenji

2006-12-01

During the onset and progression of atherosclerosis, the vascular smooth muscle cell (VSMC) phenotype changes from differentiated to dedifferentiated, and in some cases, this change is accompanied by osteogenic transition, resulting in vascular calcification. One characteristic of dedifferentiated VSMCs is the down-regulation of smooth muscle cell (SMC) marker gene expression. Bone morphogenetic proteins (BMPs), which are involved in the induction of osteogenic gene expression, are detected in calcified vasculature. In this study, we found that the BMP2-, BMP4-, and BMP6-induced expression of Msx transcription factors (Msx1 and Msx2) preceded the down-regulation of SMC marker expression in cultured differentiated VSMCs. Either Msx1 or Msx2 markedly reduced the myocardin-dependent promoter activities of SMC marker genes (SM22alpha and caldesmon). We further investigated interactions between Msx1 and myocardin/serum response factor (SRF)/CArG-box motif (cis element for SRF) using coimmunoprecipitation, gel-shift, and chromatin immunoprecipitation assays. Our results showed that Msx1 or Msx2 formed a ternary complex with SRF and myocardin and inhibited the binding of SRF or SRF/myocardin to the CArG-box motif, resulting in inhibition of their transcription.

Anabolic Properties of High Mobility Group Box Protein-1 in Human Periodontal Ligament Cells In Vitro

PubMed Central

Wolf, Michael; Lossdörfer, Stefan; Römer, Piero; Bastos Craveiro, Rogerio; Deschner, James; Jäger, Andreas

2014-01-01

High mobility group box protein-1 (HMGB1) is mainly recognized as a chemoattractant for macrophages in the initial phase of host response to pathogenic stimuli. However, recent findings provide evidence for anabolic properties in terms of enhanced proliferation, migration, and support of wound healing capacity of mesenchymal cells suggesting a dual role of the cytokine in the regulation of immune response and subsequent regenerative processes. Here, we examined potential anabolic effects of HMGB1 on human periodontal ligament (PDL) cells in the regulation of periodontal remodelling, for example, during orthodontic tooth movement. Preconfluent human PDL cells (hPDL) were exposed to HMGB1 protein and the influence on proliferation, migration, osteogenic differentiation, and biomineralization was determined by MTS assay, real time PCR, immunofluorescence cytochemistry, ELISA, and von Kossa staining. HMGB1 protein increased hPDL cell proliferation, migration, osteoblastic marker gene expression, and protein production as well as mineralized nodule formation significantly. The present findings support the dual character of HMGB1 with anabolic therapeutic potential that might support the reestablishment of the structural and functional integrity of the periodontium following periodontal trauma such as orthodontic tooth movement. PMID:25525297