Two-step interrogation then recognition of DNA binding site by Integration Host Factor: an architectural DNA-bending protein.

PubMed

Velmurugu, Yogambigai; Vivas, Paula; Connolly, Mitchell; Kuznetsov, Serguei V; Rice, Phoebe A; Ansari, Anjum

2018-02-28

The dynamics and mechanism of how site-specific DNA-bending proteins initially interrogate potential binding sites prior to recognition have remained elusive for most systems. Here we present these dynamics for Integration Host factor (IHF), a nucleoid-associated architectural protein, using a μs-resolved T-jump approach. Our studies show two distinct DNA-bending steps during site recognition by IHF. While the faster (∼100 μs) step is unaffected by changes in DNA or protein sequence that alter affinity by >100-fold, the slower (1-10 ms) step is accelerated ∼5-fold when mismatches are introduced at DNA sites that are sharply kinked in the specific complex. The amplitudes of the fast phase increase when the specific complex is destabilized and decrease with increasing [salt], which increases specificity. Taken together, these results indicate that the fast phase is non-specific DNA bending while the slow phase, which responds only to changes in DNA flexibility at the kink sites, is specific DNA kinking during site recognition. Notably, the timescales for the fast phase overlap with one-dimensional diffusion times measured for several proteins on DNA, suggesting that these dynamics reflect partial DNA bending during interrogation of potential binding sites by IHF as it scans DNA.

Site-specific recombination in the cyanobacterium Anabaena sp. strain PCC 7120 catalyzed by the integrase of coliphage HK022.

PubMed

Melnikov, Olga; Zaritsky, Arieh; Zarka, Aliza; Boussiba, Sammy; Malchin, Natalia; Yagil, Ezra; Kolot, Mikhail

2009-07-01

The integrase (Int) of the lambda-like coliphage HK022 catalyzes the site-specific integration and excision of the phage DNA into and from the chromosome of its host, Escherichia coli. Int recognizes two different pairs of recombining sites attP x attB and attL x attR for integration and excision, respectively. This system was adapted to the cyanobacterium Anabaena sp. strain PCC 7120 as a potential tool for site-specific gene manipulations in the cyanobacterium. Two plasmids were consecutively cointroduced by conjugation into Anabaena cells, one plasmid that expresses HK022 Int recombinase and the other plasmid that carries the excision substrate P(glnA)-attL-T1/T2-attR-lacZ, where T1/T2 are the strong transcription terminators of rrnB, to prevent expression of the lacZ reporter under the constitutive promoter P(glnA). The Int-catalyzed site-specific recombination reaction was monitored by the expression of lacZ emanating as a result of T1/T2 excision. Int catalyzed the site-specific excision reaction in Anabaena cells when its substrate was located either on the plasmid or on the chromosome with no need to supply an accessory protein, such as integration host factor and excisionase (Xis), which are indispensable for this reaction in its host, E. coli.

Configuration management program plan for Hanford site systems engineering

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

Kellie, C.L.

This plan establishes the integrated management program for the evolving technical baseline developed through the systems engineering process. This configuration management program aligns with the criteria identified in the DOE Standard, DOE-STD-1073-93. Included are specific requirements for control of the systems engineering RDD-100 database, and electronic data incorporated in the database that establishes the Hanford Site Technical Baseline.

Genome Integration and Excision by a New Streptomyces Bacteriophage, ϕJoe

PubMed Central

Haley, Joshua A.; Stark, W. Marshall

2016-01-01

ABSTRACT Bacteriophages are the source of many valuable tools for molecular biology and genetic manipulation. In Streptomyces, most DNA cloning vectors are based on serine integrase site-specific DNA recombination systems derived from phage. Because of their efficiency and simplicity, serine integrases are also used for diverse synthetic biology applications. Here, we present the genome of a new Streptomyces phage, ϕJoe, and investigate the conditions for integration and excision of the ϕJoe genome. ϕJoe belongs to the largest Streptomyces phage cluster (R4-like) and encodes a serine integrase. The attB site from Streptomyces venezuelae was used efficiently by an integrating plasmid, pCMF92, constructed using the ϕJoe int-attP locus. The attB site for ϕJoe integrase was occupied in several Streptomyces genomes, including that of S. coelicolor, by a mobile element that varies in gene content and size between host species. Serine integrases require a phage-encoded recombination directionality factor (RDF) to activate the excision reaction. The ϕJoe RDF was identified, and its function was confirmed in vivo. Both the integrase and RDF were active in in vitro recombination assays. The ϕJoe site-specific recombination system is likely to be an important addition to the synthetic biology and genome engineering toolbox. IMPORTANCE Streptomyces spp. are prolific producers of secondary metabolites, including many clinically useful antibiotics. Bacteriophage-derived integrases are important tools for genetic engineering, as they enable integration of heterologous DNA into the Streptomyces chromosome with ease and high efficiency. Recently, researchers have been applying phage integrases for a variety of applications in synthetic biology, including rapid assembly of novel combinations of genes, biosensors, and biocomputing. An important requirement for optimal experimental design and predictability when using integrases, however, is the need for multiple enzymes with different specificities for their integration sites. In order to provide a broad platform of integrases, we identified and validated the integrase from a newly isolated Streptomyces phage, ϕJoe. ϕJoe integrase is active in vitro and in vivo. The specific recognition site for integration is present in a wide range of different actinobacteria, including Streptomyces venezuelae, an emerging model bacterium in Streptomyces research. PMID:28003200

Configuration management program plan for Hanford site systems engineering

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

Hoffman, A.G.

This plan establishes the integrated configuration management program for the evolving technical baseline developed through the systems engineering process. This configuration management program aligns with the criteria identified in the DOE Standard, DOE-STD-1073-93. Included are specific requirements for control of the systems engineering RDD-100 database, and electronic data incorporated in the database that establishes the Hanford site technical baseline.

An integrated CRISPR Bombyx mori genome editing system with improved efficiency and expanded target sites.

PubMed

Ma, Sanyuan; Liu, Yue; Liu, Yuanyuan; Chang, Jiasong; Zhang, Tong; Wang, Xiaogang; Shi, Run; Lu, Wei; Xia, Xiaojuan; Zhao, Ping; Xia, Qingyou

2017-04-01

Genome editing enabled unprecedented new opportunities for targeted genomic engineering of a wide variety of organisms ranging from microbes, plants, animals and even human embryos. The serial establishing and rapid applications of genome editing tools significantly accelerated Bombyx mori (B. mori) research during the past years. However, the only CRISPR system in B. mori was the commonly used SpCas9, which only recognize target sites containing NGG PAM sequence. In the present study, we first improve the efficiency of our previous established SpCas9 system by 3.5 folds. The improved high efficiency was also observed at several loci in both BmNs cells and B. mori embryos. Then to expand the target sites, we showed that two newly discovered CRISPR system, SaCas9 and AsCpf1, could also induce highly efficient site-specific genome editing in BmNs cells, and constructed an integrated CRISPR system. Genome-wide analysis of targetable sites was further conducted and showed that the integrated system cover 69,144,399 sites in B. mori genome, and one site could be found in every 6.5 bp. The efficiency and resolution of this CRISPR platform will probably accelerate both fundamental researches and applicable studies in B. mori, and perhaps other insects. Copyright © 2017 Elsevier Ltd. All rights reserved.

Integrated medical and behavioral laboratory measurement system engineering analysis and laboratory specification

NASA Technical Reports Server (NTRS)

Grave, C.; Margold, D. W.

1973-01-01

Site selection, program planning, cost and design studies for support of the IMBLMS program were investigated. Accomplishments are reported for the following areas: analysis of responses to site selection criteria, space-oriented biotechnology, life sciences payload definition, and program information transfer.

Cas9-Guide RNA Directed Genome Editing in Soybean[OPEN

PubMed Central

Li, Zhongsen; Liu, Zhan-Bin; Xing, Aiqiu; Moon, Bryan P.; Koellhoffer, Jessica P.; Huang, Lingxia; Ward, R. Timothy; Clifton, Elizabeth; Falco, S. Carl; Cigan, A. Mark

2015-01-01

Recently discovered bacteria and archaea adaptive immune system consisting of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) endonuclease has been explored in targeted genome editing in different species. Streptococcus pyogenes Cas9-guide RNA (gRNA) was successfully applied to generate targeted mutagenesis, gene integration, and gene editing in soybean (Glycine max). Two genomic sites, DD20 and DD43 on chromosome 4, were mutagenized with frequencies of 59% and 76%, respectively. Sequencing randomly selected transgenic events confirmed that the genome modifications were specific to the Cas9-gRNA cleavage sites and consisted of small deletions or insertions. Targeted gene integrations through homology-directed recombination were detected by border-specific polymerase chain reaction analysis for both sites at callus stage, and one DD43 homology-directed recombination event was transmitted to T1 generation. T1 progenies of the integration event segregated according to Mendelian laws and clean homozygous T1 plants with the donor gene precisely inserted at the DD43 target site were obtained. The Cas9-gRNA system was also successfully applied to make a directed P178S mutation of acetolactate synthase1 gene through in planta gene editing. PMID:26294043

Development of a geophysical data fusion system for assessment and remediation studies of polluted groundwater aquifers. Scientific report No. 1

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

Murphy, J.R.; O`Neill, D.C.; Barker, B.W.

1994-10-01

The research described in this report is directed toward the development of a workstation-based data management, analysis and visualization system which can be used to improve the Air Force`s capability to evaluate site specific environmental hazards. The initial prototype system described in this report is directed toward a specific application to the Massachusetts Military Reservation (formerly Otis Air Force Base) on Cape Cod, Massachusetts. This system integrates a comprehensive, on-line environmental database for the site together with a map-based graphical user interface which facilitates analyst access to the databases and analysis tools needed to characterize the subsurface geologic and hydrologicmore » environments at the site.« less

Site-Specific Cassette Exchange Systems in the Aedes aegypti Mosquito and the Plutella xylostella Moth

PubMed Central

Haghighat-Khah, Roya Elaine; Scaife, Sarah; Martins, Sara; St John, Oliver; Matzen, Kelly Jean; Morrison, Neil; Alphey, Luke

2015-01-01

Genetically engineered insects are being evaluated as potential tools to decrease the economic and public health burden of mosquitoes and agricultural pest insects. Here we describe a new tool for the reliable and targeted genome manipulation of pest insects for research and field release using recombinase mediated cassette exchange (RMCE) mechanisms. We successfully demonstrated the established ΦC31-RMCE method in the yellow fever mosquito, Aedes aegypti, which is the first report of RMCE in mosquitoes. A new variant of this RMCE system, called iRMCE, combines the ΦC31-att integration system and Cre or FLP-mediated excision to remove extraneous sequences introduced as part of the site-specific integration process. Complete iRMCE was achieved in two important insect pests, Aedes aegypti and the diamondback moth, Plutella xylostella, demonstrating the transferability of the system across a wide phylogenetic range of insect pests. PMID:25830287

Retroviral integration: Site matters

PubMed Central

Demeulemeester, Jonas; De Rijck, Jan

2015-01-01

Here, we review genomic target site selection during retroviral integration as a multistep process in which specific biases are introduced at each level. The first asymmetries are introduced when the virus takes a specific route into the nucleus. Next, by co‐opting distinct host cofactors, the integration machinery is guided to particular chromatin contexts. As the viral integrase captures a local target nucleosome, specific contacts introduce fine‐grained biases in the integration site distribution. In vivo, the established population of proviruses is subject to both positive and negative selection, thereby continuously reshaping the integration site distribution. By affecting stochastic proviral expression as well as the mutagenic potential of the virus, integration site choice may be an inherent part of the evolutionary strategies used by different retroviruses to maximise reproductive success. PMID:26293289

Integrating emergency services in an urban health system.

PubMed

Radloff, D; Blouin, A S; Larsen, L; Kripp, M E

2000-03-01

When planning for growth and management efficiency across urban health systems, economic and market factors present significant service line challenges and opportunities. This article describes the evolutionary integration of emergency services in St John Health System, a large, religious-sponsored health care system located in Detroit, Michigan. Critical business elements, including the System's vision, mission, and economic context, are defined as the framework for site-specific and System-wide planning. The impact of managed care and market changes prompted St John's clinicians and executives to explore how integrating emergency services could create a competitive market advantage.

Characterizing site specific considerations for protecting aircraft during LGS operations at W. M. Keck Observatory

NASA Astrophysics Data System (ADS)

Stomski, Paul J., Jr.; Campbell, Randy; McCann, Kevin; Shimko, Steve

2010-07-01

W. M. Keck Observatory (WMKO) routinely operates laser guide star (LGS) Adaptive Optics (AO) systems at the telescope facility on the Big Island of Hawaii. One of the operational requirements for the LGS system is that a safety system to prevent nearby aircraft from being adversely affected by the laser must be provided. We will support operations in the near term with human aircraft spotters until we can successfully develop and get the appropriate approvals needed for an Automated, Integrated and Reliable System for an Aircraft Friendly Environment (AIRSAFE). This report describes some of the preliminary requirements development work at WMKO in support of the future development of AIRSAFE. We discuss the results of recent work to characterize site specific considerations that impact requirements development. The site specific considerations include the proximity of WMKO laser operations to nearby commercial airports, the implications of military operations in the area and the character of the air traffic volume and flight patterns over the telescope facility. Finally, we discuss how the design and implementation of AIRSAFE will be impacted by these site specific considerations.

Safe Genetic Modification of Cardiac Stem Cells Using a Site-Specific Integration Technique

PubMed Central

Lan, Feng; Liu, Junwei; Narsinh, Kazim H.; Hu, Shijun; Han, Leng; Lee, Andrew S.; Karow, Marisa; Nguyen, Patricia K.; Nag, Divya; Calos, Michele P.; Robbins, Robert C.; Wu, Joseph C.

2012-01-01

Background Human cardiac progenitor cells (hCPCs) are a promising cell source for regenerative repair after myocardial infarction. Exploitation of their full therapeutic potential may require stable genetic modification of the cells ex vivo. Safe genetic engineering of stem cells, using facile methods for site-specific integration of transgenes into known genomic contexts, would significantly enhance the overall safety and efficacy of cellular therapy in a variety of clinical contexts. Methods and Results We employed the phiC31 site-specific recombinase to achieve targeted integration of a triple fusion reporter gene into a known chromosomal context in hCPCs and human endothelial cells (hECs). Stable expression of the reporter gene from its unique chromosomal integration site resulted in no discernible genomic instability or adverse changes in cell phenotype. Namely, phiC31-modified hCPCs were unchanged in their differentiation propensity, cellular proliferative rate, and global gene expression profile when compared to unaltered control hCPCs. Expression of the triple fusion reporter gene enabled multimodal assessment of cell fate in vitro and in vivo using fluorescence microscopy, bioluminescence imaging (BLI), and positron emission tomography (PET). Intramyocardial transplantation of genetically modified hCPCs resulted in significant improvement in myocardial function two weeks after cell delivery, as assessed by echocardiography (P = 0.002) and magnetic resonance imaging (P = 0.001). We also demonstrated the feasibility and therapeutic efficacy of genetically modifying differentiated hECs, which enhanced hindlimb perfusion (P<0.05 at day 7 and 14 after transplantation) on laser Doppler imaging. Conclusions The phiC31 integrase genomic modification system is a safe, efficient tool to enable site-specific integration of reporter transgenes in progenitor and differentiated cell types. PMID:22965984

Safe genetic modification of cardiac stem cells using a site-specific integration technique.

PubMed

Lan, Feng; Liu, Junwei; Narsinh, Kazim H; Hu, Shijun; Han, Leng; Lee, Andrew S; Karow, Marisa; Nguyen, Patricia K; Nag, Divya; Calos, Michele P; Robbins, Robert C; Wu, Joseph C

2012-09-11

Human cardiac progenitor cells (hCPCs) are a promising cell source for regenerative repair after myocardial infarction. Exploitation of their full therapeutic potential may require stable genetic modification of the cells ex vivo. Safe genetic engineering of stem cells, using facile methods for site-specific integration of transgenes into known genomic contexts, would significantly enhance the overall safety and efficacy of cellular therapy in a variety of clinical contexts. We used the phiC31 site-specific recombinase to achieve targeted integration of a triple fusion reporter gene into a known chromosomal context in hCPCs and human endothelial cells. Stable expression of the reporter gene from its unique chromosomal integration site resulted in no discernible genomic instability or adverse changes in cell phenotype. Namely, phiC31-modified hCPCs were unchanged in their differentiation propensity, cellular proliferative rate, and global gene expression profile when compared with unaltered control hCPCs. Expression of the triple fusion reporter gene enabled multimodal assessment of cell fate in vitro and in vivo using fluorescence microscopy, bioluminescence imaging, and positron emission tomography. Intramyocardial transplantation of genetically modified hCPCs resulted in significant improvement in myocardial function 2 weeks after cell delivery, as assessed by echocardiography (P=0.002) and MRI (P=0.001). We also demonstrated the feasibility and therapeutic efficacy of genetically modifying differentiated human endothelial cells, which enhanced hind limb perfusion (P<0.05 at day 7 and 14 after transplantation) on laser Doppler imaging. The phiC31 integrase genomic modification system is a safe, efficient tool to enable site-specific integration of reporter transgenes in progenitor and differentiated cell types.

Genome Integration and Excision by a New Streptomyces Bacteriophage, ϕJoe.

PubMed

Fogg, Paul C M; Haley, Joshua A; Stark, W Marshall; Smith, Margaret C M

2017-03-01

Bacteriophages are the source of many valuable tools for molecular biology and genetic manipulation. In Streptomyces , most DNA cloning vectors are based on serine integrase site-specific DNA recombination systems derived from phage. Because of their efficiency and simplicity, serine integrases are also used for diverse synthetic biology applications. Here, we present the genome of a new Streptomyces phage, ϕJoe, and investigate the conditions for integration and excision of the ϕJoe genome. ϕJoe belongs to the largest Streptomyces phage cluster (R4-like) and encodes a serine integrase. The attB site from Streptomyces venezuelae was used efficiently by an integrating plasmid, pCMF92, constructed using the ϕJoe int-attP locus. The attB site for ϕJoe integrase was occupied in several Streptomyces genomes, including that of S. coelicolor , by a mobile element that varies in gene content and size between host species. Serine integrases require a phage-encoded recombination directionality factor (RDF) to activate the excision reaction. The ϕJoe RDF was identified, and its function was confirmed in vivo Both the integrase and RDF were active in in vitro recombination assays. The ϕJoe site-specific recombination system is likely to be an important addition to the synthetic biology and genome engineering toolbox. IMPORTANCE Streptomyces spp. are prolific producers of secondary metabolites, including many clinically useful antibiotics. Bacteriophage-derived integrases are important tools for genetic engineering, as they enable integration of heterologous DNA into the Streptomyces chromosome with ease and high efficiency. Recently, researchers have been applying phage integrases for a variety of applications in synthetic biology, including rapid assembly of novel combinations of genes, biosensors, and biocomputing. An important requirement for optimal experimental design and predictability when using integrases, however, is the need for multiple enzymes with different specificities for their integration sites. In order to provide a broad platform of integrases, we identified and validated the integrase from a newly isolated Streptomyces phage, ϕJoe. ϕJoe integrase is active in vitro and in vivo The specific recognition site for integration is present in a wide range of different actinobacteria, including Streptomyces venezuelae , an emerging model bacterium in Streptomyces research. Copyright © 2017 Fogg et al.

Construction of transformed, cultured silkworm cells and transgenic silkworm using the site-specific integrase system from phage φC31.

PubMed

Yin, Yajuan; Cao, Guangli; Xue, Renyu; Gong, Chengliang

2014-10-01

The Streptomyces bacteriophage, φC31, uses a site-specific integrase enzyme to perform efficient recombination. The recombination system uses specific sequences to integrate exogenous DNA from the phage into a host. The sequences are known as the attP site in the phage and the attB site in the host. The system can be used as a genetic manipulation tool. In this study it has been applied to the transformation of cultured BmN cells and the construction of transgenic Bombyx mori individuals. A plasmid, pSK-attB/Pie1-EGFP/Zeo-PASV40, containing a cassette designed to express a egfp-zeocin fusion gene, was co-transfected into cultured BmN cells with a helper plasmid, pSK-Pie1/NLS-Int/NSL. Expression of the egfp-zeocin fusion gene was driven by an ie-1 promoter, downstream of a φC31 attB site. The helper plasmid encoded the φC31 integrase enzyme, which was flanked by two nuclear localization signals. Expression of the egfp-zeocin fusion gene could be observed in transformed cells. The two plasmids were also transferred into silkworm eggs to obtain transgenic silkworms. Successful integration of the fusion gene was indicated by the detection of green fluorescence, which was emitted by the silkworms. Nucleotide sequence analysis demonstrated that the attB site had been cut, to allow recombination between the attB and endogenous pseudo attP sites in the cultured silkworm cells and silkworm individuals.

ANAEROBIC TREATMENT OF SOIL WASH FLUIDS FROM A WOOD PRESERVING SITE

EPA Science Inventory

An integrated system has been developed to remediate sols contaminated with pentachlorophenol (PCP) and polycyclic aromatic hydrocarbons (PAHs). This system involves the coupling of two treatment technologies, soil solvent washing and anaerobic biotreatment of the extract. Specif...

HIV promoter integration site primarily modulates transcriptional burst size rather than frequency.

PubMed

Skupsky, Ron; Burnett, John C; Foley, Jonathan E; Schaffer, David V; Arkin, Adam P

2010-09-30

Mammalian gene expression patterns, and their variability across populations of cells, are regulated by factors specific to each gene in concert with its surrounding cellular and genomic environment. Lentiviruses such as HIV integrate their genomes into semi-random genomic locations in the cells they infect, and the resulting viral gene expression provides a natural system to dissect the contributions of genomic environment to transcriptional regulation. Previously, we showed that expression heterogeneity and its modulation by specific host factors at HIV integration sites are key determinants of infected-cell fate and a possible source of latent infections. Here, we assess the integration context dependence of expression heterogeneity from diverse single integrations of a HIV-promoter/GFP-reporter cassette in Jurkat T-cells. Systematically fitting a stochastic model of gene expression to our data reveals an underlying transcriptional dynamic, by which multiple transcripts are produced during short, infrequent bursts, that quantitatively accounts for the wide, highly skewed protein expression distributions observed in each of our clonal cell populations. Interestingly, we find that the size of transcriptional bursts is the primary systematic covariate over integration sites, varying from a few to tens of transcripts across integration sites, and correlating well with mean expression. In contrast, burst frequencies are scattered about a typical value of several per cell-division time and demonstrate little correlation with the clonal means. This pattern of modulation generates consistently noisy distributions over the sampled integration positions, with large expression variability relative to the mean maintained even for the most productive integrations, and could contribute to specifying heterogeneous, integration-site-dependent viral production patterns in HIV-infected cells. Genomic environment thus emerges as a significant control parameter for gene expression variation that may contribute to structuring mammalian genomes, as well as be exploited for survival by integrating viruses.

Collection, processing and dissemination of data for the national solar demonstration program

NASA Technical Reports Server (NTRS)

Day, R. E.; Murphy, L. J.; Smok, J. T.

1978-01-01

A national solar data system developed for the DOE by IBM provides for automatic gathering, conversion, transfer, and analysis of demonstration site data. NASA requirements for this system include providing solar site hardware, engineering, data collection, and analysis. The specific tasks include: (1) solar energy system design/integration; (2) developing a site data acquisition subsystem; (3) developing a central data processing system; (4) operating the test facility at Marshall Space Flight Center; (5) collecting and analyzing data. The systematic analysis and evaluation of the data from the National Solar Data System is reflected in a monthly performance report and a solar energy system performance evaluation report.

A site-specific genetic modification for induction of pluripotency and subsequent isolation of derived lung alveolar epithelial type II cells.

PubMed

Yan, Qing; Quan, Yuan; Sun, Huanhuan; Peng, Xinmiao; Zou, Zhengyun; Alcorn, Joseph L; Wetsel, Rick A; Wang, Dachun

2014-02-01

Human induced pluripotent stem cells (hiPSCs) have great therapeutic potential in repairing defective lung alveoli. However, genetic abnormalities caused by vector integrations and low efficiency in generating hiPSCs, as well as difficulty in obtaining transplantable hiPSC-derived cell types are still major obstacles. Here we report a novel strategy using a single nonviral site-specific targeting vector with a combination of Tet-On inducible gene expression system, Cre/lox P switching gene expression system, and alveolar epithelial type II cell (ATIIC)-specific Neomycin(R) transgene expression system. With this strategy, a single copy of all of the required transgenes can be specifically knocked into a site immediately downstream of β-2-microglobulin (B2M) gene locus at a high frequency, without causing B2M dysfunction. Thus, the expression of reprogramming factors, Oct4, Sox2, cMyc, and Klf4, can be precisely regulated for efficient reprogramming of somatic cells into random integration-free or genetic mutation-free hiPSCs. The exogenous reprogramming factor transgenes can be subsequently removed after reprogramming by transient expression of Cre recombinase, and the resulting random integration-free and exogenous reprogramming factor-free hiPSCs can be selectively differentiated into a homogenous population of ATIICs. In addition, we show that these hiPSC-derived ATIICs exhibit ultrastructural characteristics and biological functions of normal ATIICs. When transplanted into bleomycin-challenged mice lungs, hiPSC-derived ATIICs efficiently remain and re-epithelialize injured alveoli to restore pulmonary function, preventing lung fibrosis and increasing survival without tumorigenic side effect. This strategy allows for the first time efficient generation of patient-specific ATIICs for possible future clinical applications. © 2013 AlphaMed Press.

A site-specific genetic modification for induction of pluripotency and subsequent isolation of derived lung alveolar epithelial type II cells

PubMed Central

Yan, Qing; Quan, Yuan; Sun, Huanhuan; Peng, Xinmiao; Zou, Zhengyun; Alcorn, Joseph L.; Wetsel, Rick A.; Wang, Dachun

2013-01-01

Human induced pluripotent stem cells (hiPSCs) have great therapeutic potential in repairing defective lung alveoli. However, genetic abnormalities caused by vector-integrations and low efficiency in generating hiPSCs, as well as difficulty in obtaining transplantable hiPSC-derived cell types, are still major obstacles. Here we report a novel strategy using a single non-viral site-specific-targeting vector with a combination of Tet-On inducible gene expression system, Cre/lox P switching gene expression system, and alveolar epithelial type II cell (ATIIC)-specific NeomycinR trangene expression system. With this strategy, a single copy of all of the required transgenes can be specifically knocked into a site immediately downstream of beta-2-microglobulin (B2M) gene locus at a high frequency, without causing B2M dysfunction. Thus, the expression of reprogramming factors, Oct4, Sox2, cMyc and Klf4, can be precisely regulated for efficient reprogramming of somatic cells into random-integration-free or genetic mutation-free hiPSCs. The exogenous reprogramming factor transgenes can be subsequently removed after reprogramming by transient expression of Cre recombinase, and the resulting random-integration-free and exogenous reprogramming-factor-free hiPSCs can be selectively differentiated into a homogenous population of ATIICs. In addition, we show that these hiPSC-derived ATIICs exhibit ultra-structural characteristics and biological functions of normal ATIICs. When transplanted into bleomycin-challenged mice lungs, hiPSC-derived ATIICs efficiently remain and re-epithelialize injured alveoli to restore pulmonary function, preventing lung fibrosis and increasing survival without tumorigenic side effect. This strategy allows for the first time efficient generation of patient-specific ATIICs for possible future clinical applications. PMID:24123810

Simultaneous and Sequential Integration by Cre/loxP Site-Specific Recombination in Saccharomyces cerevisiae.

PubMed

Choi, Ho-Jung; Kim, Yeon-Hee

2018-05-28

A Cre/ loxP -δ-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae . To allow repeated integrations, the reusable Candida glabrata MARKER ( CgMARKER ) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and β-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.

Application of the Cre/loxP Site-Specific Recombination System for Gene Transformation in Aurantiochytrium limacinum.

PubMed

Sun, Hengyi; Chen, Hao; Zang, Xiaonan; Hou, Pan; Zhou, Bingbing; Liu, Yuantao; Wu, Fei; Cao, Xiaofei; Zhang, Xuecheng

2015-06-01

The Cre/loxP site-specific recombination system was applied to Aurantiochytrium limacinum to obtain a transformant without the antibiotic resistance marker gene. First, the enhanced green fluorescent protein gene (egfp) and chloramphenicol resistance gene (Cmr), along with the two loxP loci, were integrated into the genome of A. limacinum OUC88 using 18S rDNA sequences as the homologous recombination sites. Then plasmid pSH65, containing a zeocin resistance gene (Bler) was transferred into A. limacinum OUC_CG. After induction with galactose, repeated passage in culture and PCR-based assessment, the pSH65 plasmid was lost and A. limacinum OUC_EG host was shown to no longer have resistance to 100 mg chloramphenicol/L or 5 mg zeocin/L. Through southern blotting and fluorescence detection, egfp was found to be integrated into the genome of A. limacinum OUC_EG, and EGFP was successfully expressed in the cells. The successful application of the Cre/loxP system demonstrates an experimental basis for genetic modification of A. limacinum so as to obtain transformed strains with no antibiotic resistance marker genes.

oPOSSUM: integrated tools for analysis of regulatory motif over-representation

PubMed Central

Ho Sui, Shannan J.; Fulton, Debra L.; Arenillas, David J.; Kwon, Andrew T.; Wasserman, Wyeth W.

2007-01-01

The identification of over-represented transcription factor binding sites from sets of co-expressed genes provides insights into the mechanisms of regulation for diverse biological contexts. oPOSSUM, an internet-based system for such studies of regulation, has been improved and expanded in this new release. New features include a worm-specific version for investigating binding sites conserved between Caenorhabditis elegans and C. briggsae, as well as a yeast-specific version for the analysis of co-expressed sets of Saccharomyces cerevisiae genes. The human and mouse applications feature improvements in ortholog mapping, sequence alignments and the delineation of multiple alternative promoters. oPOSSUM2, introduced for the analysis of over-represented combinations of motifs in human and mouse genes, has been integrated with the original oPOSSUM system. Analysis using user-defined background gene sets is now supported. The transcription factor binding site models have been updated to include new profiles from the JASPAR database. oPOSSUM is available at http://www.cisreg.ca/oPOSSUM/ PMID:17576675

Integration of intracardiac echocardiography and computed tomography during atrial fibrillation ablation: Combining ultrasound contours obtained from the right atrium and ventricular outflow tract.

PubMed

Nakamura, Kohki; Naito, Shigeto; Kaseno, Kenichi; Nakatani, Yosuke; Sasaki, Takehito; Anjo, Naofumi; Yamashita, Eiji; Kumagai, Koji; Funabashi, Nobusada; Kobayashi, Yoshio; Oshima, Shigeru

2017-02-01

We aimed to optimize the acquisition of the left atrial (LA) and pulmonary vein (PV) ultrasound contours for more accurate integration of intracardiac echocardiography (ICE) and computed tomography (CT) using the CARTO ® 3 system during atrial fibrillation (AF) ablation. Eighty-five AF patients underwent integration of ICE and CT using (1) the LA roof and posterior wall contours acquired from the right atrium (RA), (2) all LA/PV contours from the RA (Whole-RA-integration), (3) the LA roof/posterior wall contours from the RA and right ventricular outflow tract (RVOT) (Posterior-RA/RV-integration), and (4) all LA/PV contours from the RA and RVOT (Whole-RA/RV-integration). The integration accuracy was compared using the (1) surface registration error, (2) distances between the three-dimensional CT and eight specific sites on the anterior, posterior, superior, and inferior aspects of the right and left circumferential PV isolation lines, and (3) registration score: a score of 0 or 1 was assigned for whether or not each specific site was visually aligned with the CT, and summed for each method (0 best, 8 worst). Posterior-RA/RV-integration revealed a significantly lower surface registration error (1.30±0.15mm) than Whole-RA- and Whole-RA/RV-integration (p<0.001). The mean distances of the eight specific sites and the registration score for Posterior-RA/RV-integration (median 1.26mm and 2, respectively) were significantly smaller than those for the other integration approaches (p<0.001). Image integration with the LA roof and posterior wall contours acquired from the RA and RVOT may provide greater accuracy for catheter navigation with three-dimensional CT during AF ablation. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

One- and two-photon states for quantum information

NASA Astrophysics Data System (ADS)

Peters, Nicholas A.

To find expression stability among transgenic lines, the Recombinase Mediated Transgene Integration (RMTI) technology using the Cre/ lox-mediated site-specific gene integration system was used. The objectives were to develop an efficient method of site-specific transgene integration and to test the effectiveness of this method by assaying transgene expression in the RMTI lines. The RMTI technology allows the precise integration of a transgene in a previously placed target genomic location containing a lox site. The efficiency of CRE-mediated site-specific integration in rice by particle bombardment was found to vary from 3 to 28% in nine different experiments. Some hemizygous site-specific integration plants that were derived from homozygous target locus were found to undergo CRE-mediated reversion of the integration locus. No reversion was observed in callus; however, reverting cells may have been excluded due to selection pressure. The expression of the transgene gus was studied in all 40 callus lines, 12 regenerated T0 plants and the T1 and T2 progenies of 5 lines. The isogenic SC lines had an average expression level based on the activity of beta-glucuronidase of 158 +/- 9 units/mg protein (mean +/- SEM; n=3; variance within SC lines are expressed as standard error of the mean SEM) indicating a significantly higher level of expression, as compared to MC lines that had a much lower expression level 44 +/- 8 units/mg protein (mean +/- SEM; n=3) and the imprecise lines that had 22 +/- 8 units/mg protein (mean +/- SEM; n=3). Transgene expression in the callus cells of precise single copy lines varied by ˜3 fold, whereas that in multi-copy lines varied by ˜30 fold. Furthermore, precise single copy lines, on an average, contained ˜3.5 fold higher expression than multi-copy lines. Transgene expression in the plants of precise single-copy lines was highly variable, which was found to be due to the loss of the integration because of CRE-mediated reversion in the locus. (Abstract shortened by UMI.)

Packaging of Human Chromosome 19-Specific Adeno-Associated Virus (AAV) Integration Sites in AAV Virions during AAV Wild-Type and Recombinant AAV Vector Production

PubMed Central

Hüser, Daniela; Weger, Stefan; Heilbronn, Regine

2003-01-01

Adeno-associated virus type 2 (AAV-2) establishes latency by site-specific integration into a unique locus on human chromosome 19, called AAVS1. During the development of a sensitive real-time PCR assay for site-specific integration, AAV-AAVS1 junctions were reproducibly detected in highly purified AAV wild-type and recombinant AAV vector stocks. A series of controls documented that the junctions were packaged in AAV capsids and were newly generated during a single round of AAV production. Cloned junctions displayed variable AAV sequences fused to AAVS1. These data suggest that packaged junctions represent footprints of AAV integration during productive infection. Apparently, AAV latency established by site-specific integration and the helper virus-dependent, productive AAV cycle are more closely related than previously thought. PMID:12663794

Transgene manipulation in zebrafish by using recombinases.

PubMed

Dong, Jie; Stuart, Gary W

2004-01-01

Although much remains to be done, our results to date suggest that efficient and precise genome engineering in zebrafish will be possible in the future by using Cre recombinase and SB transposase in combination with their respective target sites. In this study, we provide the first evidence that Cre recombinase can mediate effective site-specific deletion of transgenes in zebrafish. We found that the efficiency of target site utilization could approach 100%, independent of whether the target site was provided transiently by injection or stably within an integrated transgene. Microinjection of Cre mRNA appeared to be slightly more effective for this purpose than microinjection of Cre-expressing plasmid DNA. Our work has not yet progressed to the point where SB-mediated mobilization of our transgene constructs would be observed. However, a recent report has demonstrated that SB can enhance transgenesis rates sixfold over conventional methods by efficiently mediating multiple single-copy insertion of transgenes into the zebrafish genome (Davidson et al., 2003). Therefore, it seems likely that a combined system should eventually allow both SB-mediated transgene mobilization and Cre-mediated transgene modification. Our goal is to validate methods for the precise reengineering of the zebrafish genome by using a combination of Cre-loxP and SB transposon systems. These methods can be used to delete, replace, or mobilize large pieces of DNA or to modify the genome only when and where required by the investigator. For example, it should be possible to deliver particular RNAi genes to well-expressed chromosomal loci and then exchange them easily with alternative RNAi genes for the specific suppression of alternative targets. As a nonviral vector for gene therapy, the transposon component allows for the possibility of highly efficient integration, whereas the Cre-loxP component can target the integration and/or exchange of foreign DNA into specific sites within the genome. The specificity and efficiency of this system also make it ideal for applications in which precise genome modifications are required (e.g., stock improvement). Future work should establish whether alternative recombination systems (e.g., phiC31 integrase) can improve the utility of this system. After the fish system is fully established, it would be interesting to explore its application to genome engineering in other organisms.

10 CFR 73.55 - Requirements for physical protection of licensed activities in nuclear power reactors against...

Code of Federal Regulations, 2011 CFR

2011-01-01

... integration of systems, technologies, programs, equipment, supporting processes, and implementing procedures...-in-depth methodologies to minimize the potential for an insider to adversely affect, either directly... protection of digital computer and communication systems and networks. (ii) Site-specific conditions that...

TU-A-201-01: Introduction to In-Room Imaging System Characteristics

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

Chang, J.

2016-06-15

Recent years have seen a widespread proliferation of available in-room image guidance systems for radiation therapy target localization with many centers having multiple in-room options. In this session, available imaging systems for in-room IGRT will be reviewed highlighting the main differences in workflow efficiency, targeting accuracy and image quality as it relates to target visualization. Decision-making strategies for integrating these tools into clinical image guidance protocols that are tailored to specific disease sites like H&N, lung, pelvis, and spine SBRT will be discussed. Learning Objectives: Major system characteristics of a wide range of available in-room imaging systems for IGRT. Advantagesmore » / disadvantages of different systems for site-specific IGRT considerations. Concepts of targeting accuracy and time efficiency in designing clinical imaging protocols.« less

A Decision Support System for Optimum Use of Fertilizers

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

R. L. Hoskinson; J. R. Hess; R. K. Fink

1999-07-01

The Decision Support System for Agriculture (DSS4Ag) is an expert system being developed by the Site-Specific Technologies for Agriculture (SST4Ag) precision farming research project at the INEEL. DSS4Ag uses state-of-the-art artificial intelligence and computer science technologies to make spatially variable, site-specific, economically optimum decisions on fertilizer use. The DSS4Ag has an open architecture that allows for external input and addition of new requirements and integrates its results with existing agricultural systems' infrastructures. The DSS4Ag reflects a paradigm shift in the information revolution in agriculture that is precision farming. We depict this information revolution in agriculture as an historic trend inmore » the agricultural decision-making process.« less

CRISPR-Cas and Contact-Dependent Secretion Systems Present on Excisable Pathogenicity Islands with Conserved Recombination Modules.

PubMed

Carpenter, Megan R; Kalburge, Sai S; Borowski, Joseph D; Peters, Molly C; Colwell, Rita R; Boyd, E Fidelma

2017-05-15

Pathogenicity islands (PAIs) are mobile integrated genetic elements that contain a diverse range of virulence factors. PAIs integrate into the host chromosome at a tRNA locus that contains their specific bacterial attachment site, attB , via integrase-mediated site-specific recombination generating attL and attR sites. We identified conserved recombination modules (integrases and att sites) previously described in choleragenic Vibrio cholerae PAIs but with novel cargo genes. Clustered regularly interspaced short palindromic repeat (CRISPR)-associated proteins (Cas proteins) and a type VI secretion system (T6SS) gene cluster were identified at the Vibrio pathogenicity island 1 (VPI-1) insertion site in 19 V. cholerae strains and contained the same recombination module. Two divergent type I-F CRISPR-Cas systems were identified, which differed in Cas protein homology and content. The CRISPR repeat sequence was identical among all V. cholerae strains, but the CRISPR spacer sequences and the number of spacers varied. In silico analysis suggests that the CRISPR-Cas systems were active against phages and plasmids. A type III secretion system (T3SS) was present in 12 V. cholerae strains on a 68-kb island inserted at the same tRNA-serine insertion site as VPI-2 and contained the same recombination module. Bioinformatics analysis showed that two divergent T3SSs exist among the strains examined. Both the CRISPR and T3SS islands excised site specifically from the bacterial chromosome as complete units, and the cognate integrases were essential for this excision. These data demonstrated that identical recombination modules that catalyze integration and excision from the chromosome can acquire diverse cargo genes, signifying a novel method of acquisition for both CRISPR-Cas systems and T3SSs. IMPORTANCE This work demonstrated the presence of CRISPR-Cas systems and T3SSs on PAIs. Our work showed that similar recombination modules can associate with different cargo genes and catalyze their incorporation into bacterial chromosomes, which could convert a strain into a pathogen with very different disease pathologies. Each island had the ability to excise from the chromosome as distinct, complete units for possible transfer. Evolutionary analysis of these regions indicates that they were acquired by horizontal transfer and that PAIs are the units of transfer. Similar to the case for phage evolution, PAIs have a modular structure where different functional regions are acquired by identical recombination modules. Copyright © 2017 American Society for Microbiology.

A systematic identification of species-specific protein succinylation sites using joint element features information.

PubMed

Hasan, Md Mehedi; Khatun, Mst Shamima; Mollah, Md Nurul Haque; Yong, Cao; Guo, Dianjing

2017-01-01

Lysine succinylation, an important type of protein posttranslational modification, plays significant roles in many cellular processes. Accurate identification of succinylation sites can facilitate our understanding about the molecular mechanism and potential roles of lysine succinylation. However, even in well-studied systems, a majority of the succinylation sites remain undetected because the traditional experimental approaches to succinylation site identification are often costly, time-consuming, and laborious. In silico approach, on the other hand, is potentially an alternative strategy to predict succinylation substrates. In this paper, a novel computational predictor SuccinSite2.0 was developed for predicting generic and species-specific protein succinylation sites. This predictor takes the composition of profile-based amino acid and orthogonal binary features, which were used to train a random forest classifier. We demonstrated that the proposed SuccinSite2.0 predictor outperformed other currently existing implementations on a complementarily independent dataset. Furthermore, the important features that make visible contributions to species-specific and cross-species-specific prediction of protein succinylation site were analyzed. The proposed predictor is anticipated to be a useful computational resource for lysine succinylation site prediction. The integrated species-specific online tool of SuccinSite2.0 is publicly accessible.

Seasonal Liza aurata tissue-specific DNA integrity in a multi-contaminated coastal lagoon (Ria de Aveiro, Portugal).

PubMed

Oliveira, M; Maria, V L; Ahmad, I; Pacheco, M; Santos, M A

2010-10-01

In this study, the DNA integrity of golden grey mullet (Liza aurata) collected in differently contaminated sites of a coastal lagoon, Ria de Aveiro (Portugal), was assessed, over the period of 1 year, using the DNA alkaline unwinding assay, in four different tissues (gill, kidney, liver and blood) and compared to a reference site. The four tissues displayed different DNA integrity basal levels, clearly affected by seasonal factors. Gill and kidney were, respectively, the most and least sensitive tissues. All sites demonstrated the capacity to interfere with DNA integrity. The sites displaying the highest and lowest DNA damage capability were, respectively, Barra (subject to naval traffic) and Vagos (contaminated with polycyclic aromatic hydrocarbons). In terms of seasonal variability, autumn seems to be the more critical season (more DNA damage) unlike summer when no DNA damage was found in any tissue. Data recommend the continued monitoring of this aquatic system. Copyright © 2010 Elsevier Ltd. All rights reserved.

Analysis of the French system for persons in complex situation in regard of the implementation of a public policy aiming to integrate health and social services

PubMed Central

Somme, Dominique; de Stampa, Matthieu; Périsset, Catherine; Dupont, Olivier; Ankri, Joël; Saint-Jean, Olivier

2009-01-01

Introduction The French Alzheimer plan, states that one of its principle orientations is the implementation of ‘Homes for Integration and Autonomy for Alzheimer patients’. Alzheimer disease is taken as a model to implement integration for all persons in complex situation (handicap…). Method Seventeen experimental sites were selected in January 2009 for the first two years. They were selected by policy-makers taking into account the readiness of the local policy-makers to implement the program, the diversity of sizes of sites, the diversity of the experimentation leaders' organizational link (public administrations, not for profit organizations, private organizations) and the diversity of societal structure (rural, urban and mega-urban). The experimental sites were analyzed considering six components of integration: coordination boards, unique point of access, case-management, standardized assessment tool, individualized services plan and information system. A complete and computerized tool was specifically developed for the organizational diagnosis. Results During the session, results will be exposed. These results will give a contrasted image of the French system for persons in complex situation. The tool will be also exposed and the receptivity will be discussed. Conclusion The French Alzheimer plan gives a unique opportunity to have a portrait of the French system in regard of integration challenges.

Innovation Technologies and Applications for Coastal Archaeological sites

NASA Astrophysics Data System (ADS)

Di Iorio, A.; Biliouris, D.; Guzinski, R.; Hansen, L. B.; Bagni, M.

2015-04-01

Innovation Technologies and Applications for Coastal Archaeological sites project (ITACA) aims to develop and test a management system for underwater archaeological sites in coastal regions. The discovering and monitoring service will use innovative satellite remote sensing techniques combined with image processing algorithms. The project will develop a set of applications integrated in a system pursuing the following objectives: - Search and location of ancient ship wrecks; - Monitoring of ship wrecks, ruins and historical artefacts that are now submerged; - Integration of resulting search and monitoring data with on-site data into a management tool for underwater sites; - Demonstration of the system's suitability for a service. High resolution synthetic aperture radar (TerraSAR-X, Cosmo-SkyMed) and multispectral satellite data (WorldView) will be combined to derive the relative bathymetry of the bottom of the sea up to the depth of 50 meters. The resulting data fusion will be processed using shape detection algorithms specific for archaeological items. The new algorithms, the physical modelling and the computational capabilities will be integrated into the Web-GIS, together with data recorded from surface (2D and 3D modelling) and from underwater surveys. Additional specific archaeological layers will be included into the WebGIS to facilitate the object identification through shape detection techniques and mapping. The system will be verified and validated through an extensive onground (sea) campaign carried out with both cutting edge technologies (side-scan sonar, multi beam echo sounder) and traditional means (professional scuba divers) in two test sites in Italy and Greece. The project is leaded by Planetek Hellas E.P.E. and include ALMA Sistemi sas for the "shape detection" and dissemination tasks, DHI-GRAS and Kell Srl for multispectral and SAR bathymetry. The complete consortium is composed by eleven partners and the project Kick-Off has been held in January 2014. The present contribution aims to present the project research achievements and finding at the mid-term review.

AAVS1-Targeted Plasmid Integration in AAV Producer Cell Lines.

PubMed

Luo, Yuxia; Frederick, Amy; Martin, John M; Scaria, Abraham; Cheng, Seng H; Armentano, Donna; Wadsworth, Samuel C; Vincent, Karen A

2017-06-01

Adeno-associated virus (AAV) producer cell lines are created via transfection of HeLaS3 cells with a single plasmid containing three components (the vector sequence, the AAV rep and cap genes, and a selectable marker gene). As this plasmid contains both the cis (Rep binding sites) and trans (Rep protein encoded by the rep gene) elements required for site-specific integration, it was predicted that plasmid integration might occur within the AAVS1 locus on human chromosome 19 (chr19). The objective of this study was to investigate whether integration in AAVS1 might be correlated with vector yield. Plasmid integration sites within several independent cell lines were assessed via Southern, fluorescence in situ hybridization (FISH) and PCR analyses. In the Southern analyses, the presence of fragments detected by both rep- and AAVS1-specific probes suggested that for several mid- and high-producing lines, plasmid DNA had integrated into the AAVS1 locus. Analysis with puroR and AAVS1-specific probes suggested that integration in AAVS1 was a more widespread phenomenon. High-producing AAV2-secreted alkaline phosphatase (SEAP) lines (masterwell 82 [MW82] and MW278) were evaluated via FISH using probes specific for the plasmid, AAVS1, and a chr19 marker. FISH analysis detected two plasmid integration sites in MW278 (neither in AAVS1), while a total of three sites were identified in MW82 (two in AAVS1). An inverse PCR assay confirmed integration within AAVS1 for several mid- and high-producing lines. In summary, the FISH, Southern, and PCR data provide evidence of site-specific integration of the plasmid within AAVS1 in several AAV producer cell lines. The data also suggest that integration in AAVS1 is a general phenomenon that is not necessarily restricted to high producers. The results also suggest that plasmid integration within the AAVS1 locus is not an absolute requirement for a high vector yield.

TU-A-201-00: Image Guidance Technologies and Management Strategies

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

NONE

2016-06-15

Recent years have seen a widespread proliferation of available in-room image guidance systems for radiation therapy target localization with many centers having multiple in-room options. In this session, available imaging systems for in-room IGRT will be reviewed highlighting the main differences in workflow efficiency, targeting accuracy and image quality as it relates to target visualization. Decision-making strategies for integrating these tools into clinical image guidance protocols that are tailored to specific disease sites like H&N, lung, pelvis, and spine SBRT will be discussed. Learning Objectives: Major system characteristics of a wide range of available in-room imaging systems for IGRT. Advantagesmore » / disadvantages of different systems for site-specific IGRT considerations. Concepts of targeting accuracy and time efficiency in designing clinical imaging protocols.« less

Site-Specific Integration of Foreign DNA into Minimal Bacterial and Human Target Sequences Mediated by a Conjugative Relaxase

PubMed Central

Agúndez, Leticia; González-Prieto, Coral; Machón, Cristina; Llosa, Matxalen

2012-01-01

Background Bacterial conjugation is a mechanism for horizontal DNA transfer between bacteria which requires cell to cell contact, usually mediated by self-transmissible plasmids. A protein known as relaxase is responsible for the processing of DNA during bacterial conjugation. TrwC, the relaxase of conjugative plasmid R388, is also able to catalyze site-specific integration of the transferred DNA into a copy of its target, the origin of transfer (oriT), present in a recipient plasmid. This reaction confers TrwC a high biotechnological potential as a tool for genomic engineering. Methodology/Principal Findings We have characterized this reaction by conjugal mobilization of a suicide plasmid to a recipient cell with an oriT-containing plasmid, selecting for the cointegrates. Proteins TrwA and IHF enhanced integration frequency. TrwC could also catalyze integration when it is expressed from the recipient cell. Both Y18 and Y26 catalytic tyrosil residues were essential to perform the reaction, while TrwC DNA helicase activity was dispensable. The target DNA could be reduced to 17 bp encompassing TrwC nicking and binding sites. Two human genomic sequences resembling the 17 bp segment were accepted as targets for TrwC-mediated site-specific integration. TrwC could also integrate the incoming DNA molecule into an oriT copy present in the recipient chromosome. Conclusions/Significance The results support a model for TrwC-mediated site-specific integration. This reaction may allow R388 to integrate into the genome of non-permissive hosts upon conjugative transfer. Also, the ability to act on target sequences present in the human genome underscores the biotechnological potential of conjugative relaxase TrwC as a site-specific integrase for genomic modification of human cells. PMID:22292089

B827 Chemical Synthhesis Project - Industrial Control System Integration - Statement of Work & Specification with Attachments 1-14

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

Wade, F. E.

The Chemical Synthesis Pilot Process at the Lawrence Livermore National Laboratory (LLNL) Site 300 827 Complex will be used to synthesize small quantities of material to support research and development. The project will modernize and increase current capabilities for chemical synthesis at LLNL. The primary objective of this project is the conversion of a non-automated hands-on process to a remoteoperation process, while providing enhanced batch process step control, stored recipe-specific parameter sets, process variable visibility, monitoring, alarm and warning handling, and comprehensive batch record data logging. This Statement of Work and Specification provides the industrial-grade process control requirements for themore » chemical synthesis batching control system, hereafter referred to as the “Control System” to be delivered by the System Integrator.« less

Cre/lox-Recombinase-Mediated Cassette Exchange for Reversible Site-Specific Genomic Targeting of the Disease Vector, Aedes aegypti.

PubMed

Häcker, Irina; Harrell Ii, Robert A; Eichner, Gerrit; Pilitt, Kristina L; O'Brochta, David A; Handler, Alfred M; Schetelig, Marc F

2017-03-07

Site-specific genome modification (SSM) is an important tool for mosquito functional genomics and comparative gene expression studies, which contribute to a better understanding of mosquito biology and are thus a key to finding new strategies to eliminate vector-borne diseases. Moreover, it allows for the creation of advanced transgenic strains for vector control programs. SSM circumvents the drawbacks of transposon-mediated transgenesis, where random transgene integration into the host genome results in insertional mutagenesis and variable position effects. We applied the Cre/lox recombinase-mediated cassette exchange (RMCE) system to Aedes aegypti, the vector of dengue, chikungunya, and Zika viruses. In this context we created four target site lines for RMCE and evaluated their fitness costs. Cre-RMCE is functional in a two-step mechanism and with good efficiency in Ae. aegypti. The advantages of Cre-RMCE over existing site-specific modification systems for Ae. aegypti, phiC31-RMCE and CRISPR, originate in the preservation of the recombination sites, which 1) allows successive modifications and rapid expansion or adaptation of existing systems by repeated targeting of the same site; and 2) provides reversibility, thus allowing the excision of undesired sequences. Thereby, Cre-RMCE complements existing genomic modification tools, adding flexibility and versatility to vector genome targeting.

Deep Borehole Disposal Safety Analysis.

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

Freeze, Geoffrey A.; Stein, Emily; Price, Laura L.

This report presents a preliminary safety analysis for the deep borehole disposal (DBD) concept, using a safety case framework. A safety case is an integrated collection of qualitative and quantitative arguments, evidence, and analyses that substantiate the safety, and the level of confidence in the safety, of a geologic repository. This safety case framework for DBD follows the outline of the elements of a safety case, and identifies the types of information that will be required to satisfy these elements. At this very preliminary phase of development, the DBD safety case focuses on the generic feasibility of the DBD concept.more » It is based on potential system designs, waste forms, engineering, and geologic conditions; however, no specific site or regulatory framework exists. It will progress to a site-specific safety case as the DBD concept advances into a site-specific phase, progressing through consent-based site selection and site investigation and characterization.« less

An integrated assessment of location-dependent scaling for microalgae biofuel production facilities

DOE PAGES

Coleman, André M.; Abodeely, Jared M.; Skaggs, Richard L.; ...

2014-06-19

Successful development of a large-scale microalgae-based biofuels industry requires comprehensive analysis and understanding of the feedstock supply chain—from facility siting and design through processing and upgrading of the feedstock to a fuel product. The evolution from pilot-scale production facilities to energy-scale operations presents many multi-disciplinary challenges, including a sustainable supply of water and nutrients, operational and infrastructure logistics, and economic competitiveness with petroleum-based fuels. These challenges are partially addressed by applying the Integrated Assessment Framework (IAF) – an integrated multi-scale modeling, analysis, and data management suite – to address key issues in developing and operating an open-pond microalgae production facility.more » This is done by analyzing how variability and uncertainty over space and through time affect feedstock production rates, and determining the site-specific “optimum” facility scale to minimize capital and operational expenses. This approach explicitly and systematically assesses the interdependence of biofuel production potential, associated resource requirements, and production system design trade-offs. To provide a baseline analysis, the IAF was applied in this paper to a set of sites in the southeastern U.S. with the potential to cumulatively produce 5 billion gallons per year. Finally, the results indicate costs can be reduced by scaling downstream processing capabilities to fit site-specific growing conditions, available and economically viable resources, and specific microalgal strains.« less

Heat recovery subsystem and overall system integration of fuel cell on-site integrated energy systems

NASA Technical Reports Server (NTRS)

Mougin, L. J.

1983-01-01

The best HVAC (heating, ventilating and air conditioning) subsystem to interface with the Engelhard fuel cell system for application in commercial buildings was determined. To accomplish this objective, the effects of several system and site specific parameters on the economic feasibility of fuel cell/HVAC systems were investigated. An energy flow diagram of a fuel cell/HVAC system is shown. The fuel cell system provides electricity for an electric water chiller and for domestic electric needs. Supplemental electricity is purchased from the utility if needed. An excess of electricity generated by the fuel cell system can be sold to the utility. The fuel cell system also provides thermal energy which can be used for absorption cooling, space heating and domestic hot water. Thermal storage can be incorporated into the system. Thermal energy is also provided by an auxiliary boiler if needed to supplement the fuel cell system output. Fuel cell/HVAC systems were analyzed with the TRACE computer program.

A Decision Support System for Optimum Use of Fertilizers

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

Hoskinson, Reed Louis; Hess, John Richard; Fink, Raymond Keith

1999-07-01

The Decision Support System for Agriculture (DSS4Ag) is an expert system being developed by the Site-Specific Technologies for Agriculture (SST4Ag) precision farming research project at the INEEL. DSS4Ag uses state-of-the-art artificial intelligence and computer science technologies to make spatially variable, site-specific, economically optimum decisions on fertilizer use. The DSS4Ag has an open architecture that allows for external input and addition of new requirements and integrates its results with existing agricultural systems’ infrastructures. The DSS4Ag reflects a paradigm shift in the information revolution in agriculture that is precision farming. We depict this information revolution in agriculture as an historic trend inmore » the agricultural decision-making process.« less

The Structure and Effectiveness of Health Systems: Exploring the Impact of System Integration in Rural China.

PubMed

Wang, Xin; Birch, Stephen; Ma, Huifen; Zhu, Weiming; Meng, Qingyue

2016-08-12

Facing the challenges of aging populations, increasing chronic diseases prevalence and health system fragmentation, there have been several pilots of integrated health systems in China. But little is known about their structure, mechanism and effectiveness. The aim of this paper is to analyze health system integration and develop recommendations for achieving integration. Huangzhong and Hualong counties in Qinghai province were studied as study sites, with only Huangzhong having implemented health system integration. Questionnaires, interviews, and health insurance records were sources of data. Social network analysis was employed to analyze integration, through structure measurement and effectiveness evaluation. Health system integration in Huangzhong is higher than in Hualong, so is system effectiveness. The patient referral network in Hualong has more "leapfrog" referrals. The information sharing networks in both counties are larger than the other types of networks. The average distance in the joint training network of Huangzhong is less than in Hualong. Meanwhile, there are deficiencies common to both systems. Both county health systems have strengths and limitations regarding system integration. The use of medical consortia in Huangzhong has contributed to system effectiveness. Future research might consider alternative more context specific models of health system integration.

Clinical service lines in integrated delivery systems: an initial framework and exploration.

PubMed

Parker, V A; Charns, M P; Young, G J

2001-01-01

The increasing pressures on integrated healthcare delivery systems (IDSs) to provide coordinated and cost-effective care focuses attention on the question of how to best integrate across multiple sites of care. One increasingly common approach to this issue is the development of clinical service lines that integrate specific bundles of services across the operating units of a system. This article presents a conceptual model of service lines and reports results from a descriptive investigation of service line development among members of the Industry Advisory Board--a research consortium comprising IDSs. The experiences of these IDSs (1) provide valuable insights into the range of organizational arrangements and implementation issues that are associated with service line management in healthcare systems and (2) suggest aspects of service line management worthy of further inquiry.

Integration of SPS with utility system networks

NASA Technical Reports Server (NTRS)

Kaupang, B. M.

1980-01-01

The integration of Satellite Power System (SPS) power in electric utility power systems is discussed. Specifically, the nature of the power output variations from the spacecraft to the rectenna, the operational characteristics of the rectenna power, and the impacts on the electric utility system from utilizing SPS power to serve part of the system load are treated. It is concluded that if RF beam control is an acceptable method for power control, and that the site distribution of SPS rectennas do not cause a very high local penetration (40 to 50%), SPS may be integrated into electric utility system with a few negative impacts. Increased regulating duty on the conventional generation, and a potential impact on system reliability for SPS penetration in excess of about 25% appear to be two areas of concern.

Heat pump centered integrated community energy systems: system development. Georgia Institute of Technology final report

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

Wade, D.W.; Trammell, B.C.; Dixit, B.S.

1979-12-01

Heat Pump Centered-Integrated Community Energy Systems (HP-ICES) show the promise of utilizing low-grade thermal energy for low-quality energy requirements such as space heating and cooling. The Heat Pump - Wastewater Heat Recovery (HP-WHR) scheme is one approach to an HP-ICES that proposes to reclaim low-grade thermal energy from a community's wastewater effluent. This report develops the concept of an HP-WHR system, evaluates the potential performance and economics of such a system, and examines the potential for application. A thermodynamic performance analysis of a hypothetical system projects an overall system Coefficient of Performance (C.O.P.) of from 2.181 to 2.264 for waste-watermore » temperatures varying from 50/sup 0/F to 80/sup 0/F. Primary energy source savings from the nationwide implementation of this system is projected to be 6.0 QUADS-fuel oil, or 8.5 QUADS - natural gas, or 29.7 QUADS - coal for the period 1980 to 2000, depending upon the type and mix of conventional space conditioning systems which could be displaced with the HP-WHR system. Site-specific HP-WHR system designs are presented for two application communities in Georgia. Performance analyses for these systems project annual cycle system C.O.P.'s of 2.049 and 2.519. Economic analysis on the basis of a life cycle cost comparison shows one site-specific system design to be cost competitive in the immediate market with conventional residential and light commercial HVAC systems. The second site-specific system design is shown through a similar economic analysis to be more costly than conventional systems due mainly to the current low energy costs for natural gas. It is anticipated that, as energy costs escalate, this HP-WHR system will also approach the threshold of economic viability.« less

Neurons with two sites of synaptic integration learn invariant representations.

PubMed

Körding, K P; König, P

2001-12-01

Neurons in mammalian cerebral cortex combine specific responses with respect to some stimulus features with invariant responses to other stimulus features. For example, in primary visual cortex, complex cells code for orientation of a contour but ignore its position to a certain degree. In higher areas, such as the inferotemporal cortex, translation-invariant, rotation-invariant, and even view point-invariant responses can be observed. Such properties are of obvious interest to artificial systems performing tasks like pattern recognition. It remains to be resolved how such response properties develop in biological systems. Here we present an unsupervised learning rule that addresses this problem. It is based on a neuron model with two sites of synaptic integration, allowing qualitatively different effects of input to basal and apical dendritic trees, respectively. Without supervision, the system learns to extract invariance properties using temporal or spatial continuity of stimuli. Furthermore, top-down information can be smoothly integrated in the same framework. Thus, this model lends a physiological implementation to approaches of unsupervised learning of invariant-response properties.

An Integrated Assessment of Location-Dependent Scaling for Microalgae Biofuel Production Facilities

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

Coleman, Andre M.; Abodeely, Jared; Skaggs, Richard

Successful development of a large-scale microalgae-based biofuels industry requires comprehensive analysis and understanding of the feedstock supply chain—from facility siting/design through processing/upgrading of the feedstock to a fuel product. The evolution from pilot-scale production facilities to energy-scale operations presents many multi-disciplinary challenges, including a sustainable supply of water and nutrients, operational and infrastructure logistics, and economic competitiveness with petroleum-based fuels. These challenges are addressed in part by applying the Integrated Assessment Framework (IAF)—an integrated multi-scale modeling, analysis, and data management suite—to address key issues in developing and operating an open-pond facility by analyzing how variability and uncertainty in space andmore » time affect algal feedstock production rates, and determining the site-specific “optimum” facility scale to minimize capital and operational expenses. This approach explicitly and systematically assesses the interdependence of biofuel production potential, associated resource requirements, and production system design trade-offs. The IAF was applied to a set of sites previously identified as having the potential to cumulatively produce 5 billion-gallons/year in the southeastern U.S. and results indicate costs can be reduced by selecting the most effective processing technology pathway and scaling downstream processing capabilities to fit site-specific growing conditions, available resources, and algal strains.« less

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

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

Guss, Adam M.; Rother, Michael; Zhang, Jun Kai

A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri P mcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strainsmore » that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR -regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.« less

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

DOE PAGES

Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; ...

2008-01-01

A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri P mcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strainsmore » that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR -regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.« less

Integration of aerial imaging and variable-rate technology for site-specific aerial herbicide application

USDA-ARS?s Scientific Manuscript database

As remote sensing and variable rate technology are becoming more available for aerial applicators, practical methodologies on effective integration of these technologies are needed for site-specific aerial applications of crop production and protection materials. The objectives of this study were to...

Development and test fuel cell powered on-site integrated total energy systems. Phase 3: Full-scale power plant development

NASA Technical Reports Server (NTRS)

Kaufman, A.

1982-01-01

The on-site system application analysis is summarized. Preparations were completed for the first test of a full-sized single cell. Emphasis of the methanol fuel processor development program shifted toward the use of commercial shell-and-tube heat exchangers. An improved method for predicting the carbon-monoxide tolerance of anode catalysts is described. Other stack support areas reported include improved ABA bipolar plate bonding technology, improved electrical measurement techniques for specification-testing of stack components, and anodic corrosion behavior of carbon materials.

Certification of highly complex safety-related systems.

PubMed

Reinert, D; Schaefer, M

1999-01-01

The BIA has now 15 years of experience with the certification of complex electronic systems for safety-related applications in the machinery sector. Using the example of machining centres this presentation will show the systematic procedure for verifying and validating control systems using Application Specific Integrated Circuits (ASICs) and microcomputers for safety functions. One section will describe the control structure of machining centres with control systems using "integrated safety." A diverse redundant architecture combined with crossmonitoring and forced dynamization is explained. In the main section the steps of the systematic certification procedure are explained showing some results of the certification of drilling machines. Specification reviews, design reviews with test case specification, statistical analysis, and walk-throughs are the analytical measures in the testing process. Systematic tests based on the test case specification, Electro Magnetic Interference (EMI), and environmental testing, and site acceptance tests on the machines are the testing measures for validation. A complex software driven system is always undergoing modification. Most of the changes are not safety-relevant but this has to be proven. A systematic procedure for certifying software modifications is presented in the last section of the paper.

Simultaneous non-contiguous deletions using large synthetic DNA and site-specific recombinases

PubMed Central

Krishnakumar, Radha; Grose, Carissa; Haft, Daniel H.; Zaveri, Jayshree; Alperovich, Nina; Gibson, Daniel G.; Merryman, Chuck; Glass, John I.

2014-01-01

Toward achieving rapid and large scale genome modification directly in a target organism, we have developed a new genome engineering strategy that uses a combination of bioinformatics aided design, large synthetic DNA and site-specific recombinases. Using Cre recombinase we swapped a target 126-kb segment of the Escherichia coli genome with a 72-kb synthetic DNA cassette, thereby effectively eliminating over 54 kb of genomic DNA from three non-contiguous regions in a single recombination event. We observed complete replacement of the native sequence with the modified synthetic sequence through the action of the Cre recombinase and no competition from homologous recombination. Because of the versatility and high-efficiency of the Cre-lox system, this method can be used in any organism where this system is functional as well as adapted to use with other highly precise genome engineering systems. Compared to present-day iterative approaches in genome engineering, we anticipate this method will greatly speed up the creation of reduced, modularized and optimized genomes through the integration of deletion analyses data, transcriptomics, synthetic biology and site-specific recombination. PMID:24914053

Transformation of apple (Malus × domestica) using mutants of apple acetolactate synthase as a selectable marker and analysis of the T-DNA integration sites.

PubMed

Yao, Jia-Long; Tomes, Sumathi; Gleave, Andrew P

2013-05-01

Apple acetolactate synthase mutants were generated by site-specific mutagenesis and successfully used as selection marker in tobacco and apple transformation. T-DNA/Apple genome junctions were analysed using genome-walking PCR and sequencing. An Agrobacterium-mediated genetic transformation system was developed for apple (Malus × domestica), using mutants of apple acetolactate synthase (ALS) as a selectable marker. Four apple ALS mutants were generated by site-specific mutagenesis and subsequently cloned under the transcriptional control of the CaMV 35S promoter and ocs 3' terminator, in a pART27-derived plant transformation vector. Three of the four mutations were found to confer resistance to the herbicide Glean(®), containing the active agent chlorsulfuron, in tobacco (Nicotiana tabacum) transformation. In apple transformation, leaf explants infected with Agrobacterium tumefaciens EHA105 containing one of the three ALS mutants resulted in the production of shoots on medium containing 2-8 μg L(-1) Glean(®), whilst uninfected wild-type explants failed to regenerate shoots or survive on medium containing 1 and 3 μg L(-1) Glean(®), respectively. Glean(®)-resistant, regenerated shoots were further multiplied and rooted on medium containing 10 μg L(-1) Glean(®). The T-DNA and apple genome-DNA junctions from eight rooted transgenic apple plants were analysed using genome-walking PCR amplification and sequencing. This analysis confirmed T-DNA integration into the apple genome, identified the genome integration sites and revealed the extent of any vector backbone integration, T-DNA rearrangements and deletions of apple genome DNA at the sites of integration.

Energy Systems Integration: NREL + HECO

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

Hawaiian Electric Companies' (HECO) customers are among the nation's fastest-adopters of solar PV systems. For HECO, the increased daytime PV generation raises feeder voltage profiles. Emerging technologies such as advanced PV inverters, battery storage, electric vehicles, and controllable loads also have an impact on voltage profiles. From the utility's perspective, it is yet unclear how to effectively manage these customer-sited resources. NREL is helping HECO understand its options by validating several voltage regulation strategies, making specific use of advanced inverters with voltage support functions, and their integration with other controllable sources.

Managing Mission-Critical Infrastructure

ERIC Educational Resources Information Center

Breeding, Marshall

2012-01-01

In the library context, they depend on sophisticated business applications specifically designed to support their work. This infrastructure consists of such components as integrated library systems, their associated online catalogs or discovery services, and self-check equipment, as well as a Web site and the various online tools and services…

PhiC31 recombination system demonstrates heritable germinal transmission of site-specific excision from the Arabidopsis genome

PubMed Central

2010-01-01

Background The large serine recombinase phiC31 from broad host range Streptomyces temperate phage, catalyzes the site-specific recombination of two recognition sites that differ in sequence, typically known as attachment sites attB and attP. Previously, we characterized the phiC31 catalytic activity and modes of action in the fission yeast Schizosaccharomyces pombe. Results In this work, the phiC31 recombinase gene was placed under the control of the Arabidopsis OXS3 promoter and introduced into Arabidopsis harboring a chromosomally integrated attB and attP-flanked target sequence. The phiC31 recombinase excised the attB and attP-flanked DNA, and the excision event was detected in subsequent generations in the absence of the phiC31 gene, indicating germinal transmission was possible. We further verified that the genomic excision was conservative and that introduction of a functional recombinase can be achieved through secondary transformation as well as manual crossing. Conclusion The phiC31 system performs site-specific recombination in germinal tissue, a prerequisite for generating stable lines with unwanted DNA removed. The precise site-specific deletion by phiC31 in planta demonstrates that the recombinase can be used to remove selectable markers or other introduced transgenes that are no longer desired and therefore can be a useful tool for genome engineering in plants. PMID:20178628

Efficacy and site-specificity of adenoviral vector integration mediated by the phage φC31 integrase.

PubMed

Robert, Marc-André; Zeng, Yue; Raymond, Benoît; Desfossé, Laurie; Mairey, Emilie; Tremblay, Jacques P; Massie, Bernard; Gilbert, Rénald

2012-12-01

Adenoviral vectors deleted of all their viral genes (helper-dependent [HD]) are efficient gene-transfer vehicles. Because transgene expression is rapidly lost in actively dividing cells, we investigated the feasibility of using phage φC31 integrase (φC31-Int) to integrate an HD carrying an attB site and the puromycin resistance gene into human cells (HeLa) and murine myoblasts (C2C12) by co-infection with a second HD-expressing φC31-Int. Because the HD genome is linear, we also investigated whether its circularization, through expression of Cre using a third HD, affects integration. Efficacy and specificity were determined by scoring the number of puromycin-resistant colonies and by sequencing integration sites. Unexpectedly, circularization of HD was unnecessary and it even reduced the integration efficacy. The maximum integration efficacy achieved was 0.5% in HeLa cells and 0.1% in C2C12 myoblasts. Up to 76% of the integration events occurred at pseudo attP sites and previously characterized hotspots were found. A small (two- to three-fold) increase in the number of γ-H2AX positive foci, accompanied by no noticeable change in γ-H2AX expression, indicated the low genotoxicity of φC31-Int. In conclusion, integration of HD mediated by φC31-Int is an attractive alternative to engineer cells, because it permits site-specific integration of large DNA fragments with low genotoxicity.

3MRA: A MULTI-MEDIA HUMAN AND ECOLOGICAL MODELING SYSTEM FOR SITE-SPECIFIC TO NATIONAL SCALE REGULATORY APPLICATIONS

EPA Science Inventory

3MRA provides a technology that fully integrates the full dimensionality of human and ecological exposure and risk assessment, thus allowing regulatory decisions a more complete expression of potential adverse health effects related to the disposal and reuse of contaminated waste...

New Insights into the Classification and Integration Specificity of Streptococcus Integrative Conjugative Elements through Extensive Genome Exploration

PubMed Central

Ambroset, Chloé; Coluzzi, Charles; Guédon, Gérard; Devignes, Marie-Dominique; Loux, Valentin; Lacroix, Thomas; Payot, Sophie; Leblond-Bourget, Nathalie

2016-01-01

Recent genome analyses suggest that integrative and conjugative elements (ICEs) are widespread in bacterial genomes and therefore play an essential role in horizontal transfer. However, only a few of these elements are precisely characterized and correctly delineated within sequenced bacterial genomes. Even though previous analysis showed the presence of ICEs in some species of Streptococci, the global prevalence and diversity of ICEs was not analyzed in this genus. In this study, we searched for ICEs in the completely sequenced genomes of 124 strains belonging to 27 streptococcal species. These exhaustive analyses revealed 105 putative ICEs and 26 slightly decayed elements whose limits were assessed and whose insertion site was identified. These ICEs were grouped in seven distinct unrelated or distantly related families, according to their conjugation modules. Integration of these streptococcal ICEs is catalyzed either by a site-specific tyrosine integrase, a low-specificity tyrosine integrase, a site-specific single serine integrase, a triplet of site-specific serine integrases or a DDE transposase. Analysis of their integration site led to the detection of 18 target-genes for streptococcal ICE insertion including eight that had not been identified previously (ftsK, guaA, lysS, mutT, rpmG, rpsI, traG, and ebfC). It also suggests that all specificities have evolved to minimize the impact of the insertion on the host. This overall analysis of streptococcal ICEs emphasizes their prevalence and diversity and demonstrates that exchanges or acquisitions of conjugation and recombination modules are frequent. PMID:26779141

New Insights into the Classification and Integration Specificity of Streptococcus Integrative Conjugative Elements through Extensive Genome Exploration.

PubMed

Ambroset, Chloé; Coluzzi, Charles; Guédon, Gérard; Devignes, Marie-Dominique; Loux, Valentin; Lacroix, Thomas; Payot, Sophie; Leblond-Bourget, Nathalie

2015-01-01

Recent genome analyses suggest that integrative and conjugative elements (ICEs) are widespread in bacterial genomes and therefore play an essential role in horizontal transfer. However, only a few of these elements are precisely characterized and correctly delineated within sequenced bacterial genomes. Even though previous analysis showed the presence of ICEs in some species of Streptococci, the global prevalence and diversity of ICEs was not analyzed in this genus. In this study, we searched for ICEs in the completely sequenced genomes of 124 strains belonging to 27 streptococcal species. These exhaustive analyses revealed 105 putative ICEs and 26 slightly decayed elements whose limits were assessed and whose insertion site was identified. These ICEs were grouped in seven distinct unrelated or distantly related families, according to their conjugation modules. Integration of these streptococcal ICEs is catalyzed either by a site-specific tyrosine integrase, a low-specificity tyrosine integrase, a site-specific single serine integrase, a triplet of site-specific serine integrases or a DDE transposase. Analysis of their integration site led to the detection of 18 target-genes for streptococcal ICE insertion including eight that had not been identified previously (ftsK, guaA, lysS, mutT, rpmG, rpsI, traG, and ebfC). It also suggests that all specificities have evolved to minimize the impact of the insertion on the host. This overall analysis of streptococcal ICEs emphasizes their prevalence and diversity and demonstrates that exchanges or acquisitions of conjugation and recombination modules are frequent.

Plasmid integration in a wide range of bacteria mediated by the integrase of Lactobacillus delbrueckii bacteriophage mv4.

PubMed Central

Auvray, F; Coddeville, M; Ritzenthaler, P; Dupont, L

1997-01-01

Bacteriophage mv4 is a temperate phage infecting Lactobacillus delbrueckii subsp. bulgaricus. During lysogenization, the phage integrates its genome into the host chromosome at the 3' end of a tRNA(Ser) gene through a site-specific recombination process (L. Dupont et al., J. Bacteriol., 177:586-595, 1995). A nonreplicative vector (pMC1) based on the mv4 integrative elements (attP site and integrase-coding int gene) is able to integrate into the chromosome of a wide range of bacterial hosts, including Lactobacillus plantarum, Lactobacillus casei (two strains), Lactococcus lactis subsp. cremoris, Enterococcus faecalis, and Streptococcus pneumoniae. Integrative recombination of pMC1 into the chromosomes of all of these species is dependent on the int gene product and occurs specifically at the pMC1 attP site. The isolation and sequencing of pMC1 integration sites from these bacteria showed that in lactobacilli, pMC1 integrated into the conserved tRNA(Ser) gene. In the other bacterial species where this tRNA gene is less or not conserved; secondary integration sites either in potential protein-coding regions or in intergenic DNA were used. A consensus sequence was deduced from the analysis of the different integration sites. The comparison of these sequences demonstrated the flexibility of the integrase for the bacterial integration site and suggested the importance of the trinucleotide CCT at the 5' end of the core in the strand exchange reaction. PMID:9068626

Development of a high efficiency integration system and promoter library for rapid modification of Pseudomonas putida KT2440

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

Elmore, Joshua R.; Furches, Anna; Wolff, Gara N.

Pseudomonas putida strains are highly robust bacteria known for their ability to efficiently utilize a variety of carbon sources, including aliphatic and aromatic hydrocarbons. Recently, P. putida has been engineered to valorize the lignin stream of a lignocellulosic biomass pretreatment process. Nonetheless, when compared to platform organisms such as Escherichia coli, the toolkit for engineering P. putida is underdeveloped. Heterologous gene expression in particular is problematic. Plasmid instability and copy number variance provide challenges for replicative plasmids, while use of homologous recombination for insertion of DNA into the chromosome is slow and laborious. Furthermore, heterologous expression efforts to date typicallymore » rely on overexpression of exogenous pathways using a handful of poorly characterized promoters. In order to improve the P. putida toolkit, we developed a rapid genome integration system using the site-specific recombinase from bacteriophage Bxb1 to enable rapid, high efficiency integration of DNA into the P. putida chromosome. We also developed a library of synthetic promoters with various UP elements, -35 sequences, and -10 sequences, as well as different ribosomal binding sites. We tested these promoters using a fluorescent reporter gene, mNeonGreen, to characterize the strength of each promoter, and identified UP-element-promoter-ribosomal binding sites combinations capable of driving a ~150-fold range of protein expression levels. One additional integrating vector was developed that confers more robust kanamycin resistance when integrated at single copy into the chromosome. This genome integration and reporter systems are extensible for testing other genetic parts, such as examining terminator strength, and will allow rapid integration of heterologous pathways for metabolic engineering.« less

Towards integrated care for chronic conditions: Dutch policy developments to overcome the (financial) barriers.

PubMed

Tsiachristas, Apostolos; Hipple-Walters, Bethany; Lemmens, Karin M M; Nieboer, Anna P; Rutten-van Mölken, Maureen P M H

2011-07-01

Chronic non-communicable diseases are a major threat to population health and have a major economic impact on health care systems. Worldwide, integrated chronic care delivery systems have been developed to tackle this challenge. In the Netherlands, the recently introduced integrated payment system--the chain-DTC--is seen as the cornerstone of a policy stimulating the development of a well-functioning integrated chronic care system. The purpose of this paper is to describe the recent attempts in the Netherlands to stimulate the delivery of integrated chronic care, focusing specifically on the new integrated payment scheme and the barriers to introducing this scheme. We also highlight possible threats and identify necessary conditions to the success of the system. This paper is based on a combination of methods and sources including literature, government documents, personal communications and site visits to disease management programs (DMPs). The most important conditions for the success of the new payment system are: complete care protocols describing both general (e.g. smoking cessation, physical activity) and disease-specific chronic care modules, coverage of all components of a DMP by basic health care insurance, adequate information systems that facilitate communication between caregivers, explicit links between the quality and the price of a DMP, expansion of the amount of specialized care included in the chain-DTC, inclusion of a multi-morbidity factor in the risk equalization formula of insurers, and thorough economic evaluation of DMPs. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

Bacteriophage-based vectors for site-specific insertion of DNA in the chromosome of Corynebacteria.

PubMed

Oram, Mark; Woolston, Joelle E; Jacobson, Andrew D; Holmes, Randall K; Oram, Diana M

2007-04-15

In Corynebacterium diphtheriae, diphtheria toxin is encoded by the tox gene of some temperate corynephages such as beta. beta-like corynephages are capable of inserting into the C. diphtheriae chromosome at two specific sites, attB1 and attB2. Transcription of the phage-encoded tox gene, and many chromosomally encoded genes, is regulated by the DtxR protein in response to Fe(2+) levels. Characterizing DtxR-dependent gene regulation is pivotal in understanding diphtheria pathogenesis and mechanisms of iron-dependent gene expression; although this has been hampered by a lack of molecular genetic tools in C. diphtheriae and related Coryneform species. To expand the systems for genetic manipulation of C. diphtheriae, we constructed plasmid vectors capable of integrating into the chromosome. These plasmids contain the beta-encoded attP site and the DIP0182 integrase gene of C. diphtheriae NCTC13129. When these vectors were delivered to the cytoplasm of non-lysogenic C. diphtheriae, they integrated into either the attB1 or attB2 sites with comparable frequency. Lysogens were also transformed with these vectors, by virtue of the second attB site. An integrated vector carrying an intact dtxR gene complemented the mutant phenotypes of a C. diphtheriae DeltadtxR strain. Additionally, strains of beta-susceptible C. ulcerans, and C. glutamicum, a species non-permissive for beta, were each transformed with these vectors. This work significantly extends the tools available for targeted transformation of both pathogenic and non-pathogenic Corynebacterium species.

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

Bower, J.C.; Burford, M.J.; Downing, T.R.

The Integrated Baseline System (IBS) is an emergency management planning and analysis tool that is being developed under the direction of the US Army Nuclear and Chemical Agency (USANCA). The IBS Data Management Guide provides the background, as well as the operations and procedures needed to generate and maintain a site-specific map database. Data and system managers use this guide to manage the data files and database that support the administrative, user-environment, database management, and operational capabilities of the IBS. This document provides a description of the data files and structures necessary for running the IBS software and using themore » site map database.« less

Scan to Bim for 3d Reconstruction of the Papal Basilica of Saint Francis in Assisi in Italy

NASA Astrophysics Data System (ADS)

Angelini, M. G.; Baiocchi, V.; Costantino, D.; Garzia, F.

2017-05-01

The historical building heritage, present in the most of Italian cities centres, is, as part of the construction sector, a working potential, but unfortunately it requires planning of more complex and problematic interventions. However, policies to support on the existing interventions, together with a growing sensitivity for the recovery of assets, determine the need to implement specific studies and to analyse the specific problems of each site. The purpose of this paper is to illustrate the methodology and the results obtained from integrated laser scanning activity in order to have precious architectural information useful not only from the cultural heritage point of view but also to construct more operative and powerful tools, such as BIM (Building Information Modelling) aimed to the management of this cultural heritage. The Papal Basilica and the Sacred Convent of Saint Francis in Assisi in Italy are, in fact, characterized by unique and complex peculiarities, which require a detailed knowledge of the sites themselves to ensure visitor's security and safety. For such a project, we have to take in account all the people and personnel normally present in the site, visitors with disabilities and finally the needs for cultural heritage preservation and protection. This aim can be reached using integrated systems and new technologies, such as Internet of Everything (IoE), capable of connecting people, things (smart sensors, devices and actuators; mobile terminals; wearable devices; etc.), data/information/knowledge and processes to reach the desired goals. The IoE system must implement and support an Integrated Multidisciplinary Model for Security and Safety Management (IMMSSM) for the specific context, using a multidisciplinary approach.

MEETING IN TUCSON: 3MRA: A MULTI-MEDIA HUMAN AND ECOLOGICAL MODELING SYSTEM FOR SITE-SPECIFIC TO NATIONAL SCALE REGULATORY APPLICATIONS

EPA Science Inventory

3MRA provides a technology that fully integrates the full dimensionality of human and ecological exposure and risk assessment, thus allowing regulatory decisions a more complete expression of potential adverse health effects related to the disposal and reuse of contaminated waste...

78 FR 19695 - National Advisory Committee on Institutional Quality and Integrity (NACIQI)

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-02

... of higher education under Title IV, of the HEA, together with recommendations for improvement in such... available via the Federal Digital System at: www.gpo.gov/fdsys . At this site you can view this document, as... the article search feature at: www.federalregister.gov . Specifically, through the advanced search...

77 FR 64493 - National Advisory Committee on Institutional Quality and Integrity (NACIQI)

Federal Register 2010, 2011, 2012, 2013, 2014

2012-10-22

... process for institutions of higher education under Title IV, of the HEA, together with recommendations for... available via the Federal Digital System at: www.gpo.gov/fdsys . At this site you can view this document, as... the article search feature at: www.federalregister.gov . Specifically, through the advanced search...

Molecular Characterization of Transgene Integration by Next-Generation Sequencing in Transgenic Cattle

PubMed Central

Zhang, Ran; Yin, Yinliang; Zhang, Yujun; Li, Kexin; Zhu, Hongxia; Gong, Qin; Wang, Jianwu; Hu, Xiaoxiang; Li, Ning

2012-01-01

As the number of transgenic livestock increases, reliable detection and molecular characterization of transgene integration sites and copy number are crucial not only for interpreting the relationship between the integration site and the specific phenotype but also for commercial and economic demands. However, the ability of conventional PCR techniques to detect incomplete and multiple integration events is limited, making it technically challenging to characterize transgenes. Next-generation sequencing has enabled cost-effective, routine and widespread high-throughput genomic analysis. Here, we demonstrate the use of next-generation sequencing to extensively characterize cattle harboring a 150-kb human lactoferrin transgene that was initially analyzed by chromosome walking without success. Using this approach, the sites upstream and downstream of the target gene integration site in the host genome were identified at the single nucleotide level. The sequencing result was verified by event-specific PCR for the integration sites and FISH for the chromosomal location. Sequencing depth analysis revealed that multiple copies of the incomplete target gene and the vector backbone were present in the host genome. Upon integration, complex recombination was also observed between the target gene and the vector backbone. These findings indicate that next-generation sequencing is a reliable and accurate approach for the molecular characterization of the transgene sequence, integration sites and copy number in transgenic species. PMID:23185606

Characterization of IntA, a Bidirectional Site-Specific Recombinase Required for Conjugative Transfer of the Symbiotic Plasmid of Rhizobium etli CFN42

PubMed Central

Hernández-Tamayo, Rogelio; Sohlenkamp, Christian; Puente, José Luis; Brom, Susana

2013-01-01

Site-specific recombination occurs at short specific sequences, mediated by the cognate recombinases. IntA is a recombinase from Rhizobium etli CFN42 and belongs to the tyrosine recombinase family. It allows cointegration of plasmid p42a and the symbiotic plasmid via site-specific recombination between attachment regions (attA and attD) located in each replicon. Cointegration is needed for conjugative transfer of the symbiotic plasmid. To characterize this system, two plasmids harboring the corresponding attachment sites and intA were constructed. Introduction of these plasmids into R. etli revealed IntA-dependent recombination events occurring at high frequency. Interestingly, IntA promotes not only integration, but also excision events, albeit at a lower frequency. Thus, R. etli IntA appears to be a bidirectional recombinase. IntA was purified and used to set up electrophoretic mobility shift assays with linear fragments containing attA and attD. IntA-dependent retarded complexes were observed only with fragments containing either attA or attD. Specific retarded complexes, as well as normal in vivo recombination abilities, were seen even in derivatives harboring only a minimal attachment region (comprising the 5-bp central region flanked by 9- to 11-bp inverted repeats). DNase I-footprinting assays with IntA revealed specific protection of these zones. Mutations that disrupt the integrity of the 9- to 11-bp inverted repeats abolish both specific binding and recombination ability, while mutations in the 5-bp central region severely reduce both binding and recombination. These results show that IntA is a bidirectional recombinase that binds to att regions without requiring neighboring sequences as enhancers of recombination. PMID:23935046

75 FR 76487 - Haldex Brake Corporation, Commercial Vehicle Systems, Including On-Site Leased Workers of...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-08

..., Commercial Vehicle Systems, Including On-Site Leased Workers of Johnston Integration Technologies, a... system components. The company reports that workers leased from Johnston Integration Technologies, a... certification to include workers leased from Johnston Integration Technologies, a subsidiary of Johnston...

Inducible model for β-six-mediated site-specific recombination in mammalian cells

PubMed Central

Servert, Pilar; Garcia-Castro, Javier; Díaz, Vicente; Lucas, Daniel; Gonzalez, Manuel A.; Martínez-A, Carlos; Bernad, Antonio

2006-01-01

The prokaryotic β recombinase catalyzes site-specific recombination between two directly oriented minimal six sites in chromatin-integrated substrates. Here, we demonstrate that an enhanced green fluorescent protein (EGFP)-fused version of β recombinase (β-EGFP) is fully active, retaining most specific activity. It is used to develop a recombination-dependent activatable gene expression (RAGE) system based on the androgen receptor (AR) ligand-binding domain (LBD). Two hybrid molecules, a direct fusion of the LBD-AR to the C-terminus of β recombinase (β-AR) and a triple fusion of β-EGFP to the same ligand-binding domain (β-EGFP-AR), were engineered and their subcellular behavior, stability and catalytic activity were evaluated. Both chimeric β recombinase proteins showed in vivo inducible recombinogenic activity dependent on addition of an androgen receptor agonist, although the β-AR fusion protein demonstrated more accurate ligand-dependent translocation from cytoplasm to nucleus. PMID:16394020

Analysis of illegitimate genomic integration mediated by zinc-finger nucleases: implications for specificity of targeted gene correction

PubMed Central

2010-01-01

Background Formation of site specific genomic double strand breaks (DSBs), induced by the expression of a pair of engineered zinc-finger nucleases (ZFNs), dramatically increases the rates of homologous recombination (HR) between a specific genomic target and a donor plasmid. However, for the safe use of ZFN induced HR in practical applications, possible adverse effects of the technology such as cytotoxicity and genotoxicity need to be well understood. In this work, off-target activity of a pair of ZFNs has been examined by measuring the ratio between HR and illegitimate genomic integration in cells that are growing exponentially, and in cells that have been arrested in the G2/M phase. Results A reporter cell line that contained consensus ZFN binding sites in an enhanced green fluorescent protein (EGFP) reporter gene was used to measure ratios between HR and non-homologous integration of a plasmid template. Both in human cells (HEK 293) containing the consensus ZFN binding sites and in cells lacking the ZFN binding sites, a 3.5 fold increase in the level of illegitimate integration was observed upon ZFN expression. Since the reporter gene containing the consensus ZFN target sites was found to be intact in cells where illegitimate integration had occurred, increased rates of illegitimate integration most likely resulted from the formation of off-target genomic DSBs. Additionally, in a fraction of the ZFN treated cells the co-occurrence of both specific HR and illegitimate integration was observed. As a mean to minimize unspecific effects, cell cycle manipulation of the target cells by induction of a transient G2/M cell cycle arrest was shown to stimulate the activity of HR while having little effect on the levels of illegitimate integration, thus resulting in a nearly eight fold increase in the ratio between the two processes. Conclusions The demonstration that ZFN expression, in addition to stimulating specific gene targeting by HR, leads to increased rates of illegitimate integration emphasizes the importance of careful characterization of ZFN treated cells. In order to reduce off-target events, reversible cell cycle arrest of the target cells in the G2/M phase is an efficient way for increasing the ratio between specific HR and illegitimate integration. PMID:20459736

Low-cost modular array-field designs for flat-panel and concentrator photovoltaic systems

NASA Astrophysics Data System (ADS)

Post, H. N.; Carmichael, D. C.; Alexander, G.; Castle, J. A.

1982-09-01

Described are the design and development of low-cost, modular array fields for flat-panel and concentrator photovoltaic (PV) systems. The objective of the work was to reduce substantially the cost of the array-field Balance-of-System (BOS) subsystems and site-specific design costs as compared to previous PV installations. These subsystems include site preparation, foundations, support structures, electrical writing, grounding, lightning protection, electromagnetic interference considerations, and controls. To reduce these BOS and design costs, standardized modular (building-block) designs for flat-panel and concentrator array fields have been developed that are fully integrated and optimized for lowest life-cycle costs. Using drawings and specifications now available, these building-block designs can be used in multiples to install various size array fields. The developed designs are immediately applicable (1982) and reduce the array-field BOS costs to a fraction of previous costs.

Assessing time-integrated dissolved concentrations and predicting toxicity of metals during diel cycling in streams

USGS Publications Warehouse

Balistrieri, Laurie S.; Nimick, David A.; Mebane, Christopher A.

2012-01-01

Evaluating water quality and the health of aquatic organisms is challenging in systems with systematic diel (24 hour) or less predictable runoff-induced changes in water composition. To advance our understanding of how to evaluate environmental health in these dynamic systems, field studies of diel cycling were conducted in two streams (Silver Bow Creek and High Ore Creek) affected by historical mining activities in southwestern Montana. A combination of sampling and modeling tools were used to assess the toxicity of metals in these systems. Diffusive Gradients in Thin Films (DGT) samplers were deployed at multiple time intervals during diel sampling to confirm that DGT integrates time-varying concentrations of dissolved metals. Thermodynamic speciation calculations using site specific water compositions, including time-integrated dissolved metal concentrations determined from DGT, and a competitive, multiple-metal biotic ligand model incorporated into the Windemere Humic Aqueous Model Version 6.0 (WHAM VI) were used to determine the chemical speciation of dissolved metals and biotic ligands. The model results were combined with previously collected toxicity data on cutthroat trout to derive a relationship that predicts the relative survivability of these fish at a given site. This integrative approach may prove useful for assessing water quality and toxicity of metals to aquatic organisms in dynamic systems and evaluating whether potential changes in environmental health of aquatic systems are due to anthropogenic activities or natural variability.

The Bxb1 recombination system demonstrates heritable transmission of site-specific excision in Arabidopsis

PubMed Central

2012-01-01

Background The mycobacteriophage large serine recombinase Bxb1 catalyzes site-specific recombination between its corresponding attP and attB recognition sites. Previously, we and others have shown that Bxb1 has catalytic activity in various eukaryotic species including Nicotiana tabacum, Schizosaccharomyces pombe, insects and mammalian cells. Results In this work, the Bxb1 recombinase gene was transformed and constitutively expressed in Arabidopsis thaliana plants harboring a chromosomally integrated attP and attB-flanked target sequence. The Bxb1 recombinase successfully excised the target sequence in a conservative manner and the resulting recombination event was heritably transmitted to subsequent generations in the absence of the recombinase transgene. In addition, we also show that Bxb1 recombinase expressing plants can be manually crossed with att-flanked target transgenic plants to generate excised progeny. Conclusion The Bxb1 large serine recombinase performs site-specific recombination in Arabidopsis thaliana germinal tissue, producing stable lines free of unwanted DNA. The precise site-specific deletion produced by Bxb1 in planta demonstrates that this enzyme can be a useful tool for the genetic engineering of plants without selectable marker transgenes or other undesirable exogenous sequences. PMID:22436504

The phage integrase vector pIPI03 allows RecA-independent, site-specific labelling of Staphylococcus lugdunensis strains.

PubMed

Heilbronner, Simon; Monk, Ian R; Foster, Timothy J

2013-11-01

Staphylococcus lugdunensis is a coagulase negative staphylococcus that is a commensal of man and an opportunistic pathogen. A site-specific integrative plasmid for the use in S. lugdunensis was constructed and validated. The integrase gene ccrB of bacteriophage ϕSL01 together with its attachment site was cloned into the thermosensitive plasmid pIMAY. The resulting plasmid pIPI03 integrated RecA-independently, site-specifically and irreversibly into the S. lugdunensis chromosome. Two IPTG-inducible antibiotic resistance determinants were cloned into pIPI03 and the derivatives were used to construct strains suitable for competitive growth experiments in both in vitro and in vivo. Copyright © 2013 Elsevier Inc. All rights reserved.

Guidelines for development of structural integrity programs for DOE high-level waste storage tanks

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

Bandyopadhyay, K.; Bush, S.; Kassir, M.

Guidelines are provided for developing programs to promote the structural integrity of high-level waste storage tanks and transfer lines at the facilities of the Department of Energy. Elements of the program plan include a leak-detection system, definition of appropriate loads, collection of data for possible material and geometric changes, assessment of the tank structure, and non-destructive examination. Possible aging degradation mechanisms are explored for both steel and concrete components of the tanks, and evaluated to screen out nonsignificant aging mechanisms and to indicate methods of controlling the significant aging mechanisms. Specific guidelines for assessing structural adequacy will be provided inmore » companion documents. Site-specific structural integrity programs can be developed drawing on the relevant portions of the material in this document.« less

Study of Systems and Technology for Liquid Hydrogen Production Independent of Fossil Fuels

NASA Technical Reports Server (NTRS)

Sprafka, R. J.; Escher, W. J. D.; Foster, R. W.; Tison, R. R.; Shingleton, J.; Moore, J. S.; Baker, C. R.

1983-01-01

Based on Kennedy Space Center siting and logistics requirements and the nonfossil energy resources at the Center, a number of applicable technologies and system candidates for hydrogen production were identified and characterized. A two stage screening of these technologies in the light of specific criteria identified two leading candidates as nonfossil system approaches. Conceptual design and costing of two solar-operated, stand alone systems, one photovoltaic based on and the other involving the power tower approach reveals their technical feasibility as sited as KSC, and the potential for product cost competitiveness with conventional supply approaches in the 1990 to 1210 time period. Conventional water hydrolysis and hydrogen liquefaction subsystems are integrated with the solar subsystems.

[Expression of human-mouse chimeric antibody directed against Chikungunya virus with site-specific integration system].

PubMed

Li, Jian-min; Chen, Wei; Jia, Xiu-jie; An, Xiao-ping; Li, Bing; Fan, Ying-ru; Tong, Yi-gang

2005-05-01

To obtain CHO/dhfr(-) cells line with integrated FRT sequence in the chromosome transcription active site and to express human-mouse chimeric antibody directed against Chikungunya Virus by using the cell line. The fusion gene of FRT and HBsAg was constructed by PCR and cloned into the MCS of pCI-neo to construct pCI-FRT-HBsAg. The pCI-FRT-HBsAg was transfected into CHO/dhfr(-) cells and cell clones with high expression of HBsAg were screened by detecting the amount of HBsAg with ELISA. A CHO cell clone with the highest expression was chosen and named as CHO/dhfr(-) FRT(+). pAFRT HFLF, a expression plasmid of chimeric antibody with RFT sequence was transfected into CHO/dhfr(-) FRT(+) cells and cell clones with high expression of the chimeric antibody were screened by increasing concentration of MTX. A CHO cell clone with high expression of the chimeric antibody was cultured in large scale and supernatant was collected from which the chimeric antibody was purified. The purified chimeric antibody was analyzed by SDS-PAGE, Western blot and IFA. A CHO/dhfr(-) cells line with integrated FRT sequence in the chromosome transcription active site was obtained successfully. A cell clone with yield of 5 mg/L of chimeric antibody was obtained, as compared with routine CHO cell expression system with a yield of 2 mg/L. A cell line with integrated FRT sequence in the chromosome transcription active site was obtained and with it human-mouse chimeric antibody directed against Chikungunya virus was expressed. This system lays a solid foundation which can be used for expressing antibodies and other proteins.

Implementing a user-driven online quality improvement toolkit for cancer care.

PubMed

Luck, Jeff; York, Laura S; Bowman, Candice; Gale, Randall C; Smith, Nina; Asch, Steven M

2015-05-01

Peer-to-peer collaboration within integrated health systems requires a mechanism for sharing quality improvement lessons. The Veterans Health Administration (VA) developed online compendia of tools linked to specific cancer quality indicators. We evaluated awareness and use of the toolkits, variation across facilities, impact of social marketing, and factors influencing toolkit use. A diffusion of innovations conceptual framework guided the collection of user activity data from the Toolkit Series SharePoint site and an online survey of potential Lung Cancer Care Toolkit users. The VA Toolkit Series site had 5,088 unique visitors in its first 22 months; 5% of users accounted for 40% of page views. Social marketing communications were correlated with site usage. Of survey respondents (n = 355), 54% had visited the site, of whom 24% downloaded at least one tool. Respondents' awareness of the lung cancer quality performance of their facility, and facility participation in quality improvement collaboratives, were positively associated with Toolkit Series site use. Facility-level lung cancer tool implementation varied widely across tool types. The VA Toolkit Series achieved widespread use and a high degree of user engagement, although use varied widely across facilities. The most active users were aware of and active in cancer care quality improvement. Toolkit use seemed to be reinforced by other quality improvement activities. A combination of user-driven tool creation and centralized toolkit development seemed to be effective for leveraging health information technology to spread disease-specific quality improvement tools within an integrated health care system. Copyright © 2015 by American Society of Clinical Oncology.

Investigating Uncertainty and Sensitivity in Integrated, Multimedia Environmental Models: Tools for FRAMES-3MRA

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

Babendreier, Justin E.; Castleton, Karl J.

2005-08-01

Elucidating uncertainty and sensitivity structures in environmental models can be a difficult task, even for low-order, single-medium constructs driven by a unique set of site-specific data. Quantitative assessment of integrated, multimedia models that simulate hundreds of sites, spanning multiple geographical and ecological regions, will ultimately require a comparative approach using several techniques, coupled with sufficient computational power. The Framework for Risk Analysis in Multimedia Environmental Systems - Multimedia, Multipathway, and Multireceptor Risk Assessment (FRAMES-3MRA) is an important software model being developed by the United States Environmental Protection Agency for use in risk assessment of hazardous waste management facilities. The 3MRAmore » modeling system includes a set of 17 science modules that collectively simulate release, fate and transport, exposure, and risk associated with hazardous contaminants disposed of in land-based waste management units (WMU) .« less

A novel site-specific recombination system derived from bacteriophage phiMR11.

PubMed

Rashel, Mohammad; Uchiyama, Jumpei; Ujihara, Takako; Takemura, Iyo; Hoshiba, Hiroshi; Matsuzaki, Shigenobu

2008-04-04

We report identification of a novel site-specific DNA recombination system that functions in both in vivo and in vitro, derived from lysogenic Staphylococcus aureus phage phiMR11. In silico analysis of the phiMR11 genome indicated orf1 as a putative integrase gene. Phage and bacterial attachment sites (attP and attB, respectively) and attachment junctions were determined and their nucleotide sequences decoded. Sequences of attP and attB were mostly different to each other except for a two bp common core that was the crossover point. We found several inverted repeats adjacent to the core sequence of attP as potential protein binding sites. The precise and efficient integration properties of phiMR11 integrase were shown on attP and attB in Escherichia coli and the minimum size of attP was found to be 34bp. In in vitro assays using crude or purified integrase, only buffer and substrate DNAs were required for the recombination reaction, indicating that other bacterially encoded factors are not essential for activity.

Dissemination of compressed satellite imagery within the Navy SPAWAR Central Site Product Display environment

NASA Technical Reports Server (NTRS)

Kiselyov, Oleg; Fisher, Paul

1995-01-01

This paper presents a case study of integration of compression techniques within a satellite image communication component of an actual tactical weather information dissemination system. The paper describes history and requirements of the project, and discusses the information flow, request/reply protocols, error handling, and, especially, system integration issues: specification of compression parameters and the place and time for compressor/decompressor plug-ins. A case for a non-uniform compression of satellite imagery is presented, and its implementation in the current system id demonstrated. The paper gives special attention to challenges of moving the system towards the use of standard, non-proprietary protocols (smtp and http) and new technologies (OpenDoc), and reports the ongoing work in this direction.

CLIPS: A tool for the development and delivery of expert systems

NASA Technical Reports Server (NTRS)

Riley, Gary

1991-01-01

The C Language Integrated Production System (CLIPS) is a forward chaining rule-based language developed by the Software Technology Branch at the Johnson Space Center. CLIPS provides a complete environment for the construction of rule-based expert systems. CLIPS was designed specifically to provide high probability, low cost, and easy integration with external systems. Other key features of CLIPS include a powerful rule syntax, an interactive development environment, high performance, extensibility, a verification/validation tool, extensive documentation, and source code availability. The current release of CLIPS, version 4.3, is being used by over 2,500 users throughout the public and private community including: all NASA sites and branches of the military, numerous Federal bureaus, government contractors, 140 universities, and many companies.

Rapid development of stable transgene CHO cell lines by CRISPR/Cas9-mediated site-specific integration into C12orf35.

PubMed

Zhao, Menglin; Wang, Jiaxian; Luo, Manyu; Luo, Han; Zhao, Meiqi; Han, Lei; Zhang, Mengxiao; Yang, Hui; Xie, Yueqing; Jiang, Hua; Feng, Lei; Lu, Huili; Zhu, Jianwei

2018-07-01

Chinese hamster ovary (CHO) cells are the most widely used mammalian hosts for recombinant protein production. However, by conventional random integration strategy, development of a high-expressing and stable recombinant CHO cell line has always been a difficult task due to the heterogenic insertion and its caused requirement of multiple rounds of selection. Site-specific integration of transgenes into CHO hot spots is an ideal strategy to overcome these challenges since it can generate isogenic cell lines with consistent productivity and stability. In this study, we investigated three sites with potential high transcriptional activities: C12orf35, HPRT, and GRIK1, to determine the possible transcriptional hot spots in CHO cells, and further construct a reliable site-specific integration strategy to develop recombinant cell lines efficiently. Genes encoding representative proteins mCherry and anti-PD1 monoclonal antibody were targeted into these three loci respectively through CRISPR/Cas9 technology. Stable cell lines were generated successfully after a single round of selection. In comparison with a random integration control, all the targeted integration cell lines showed higher productivity, among which C12orf35 locus was the most advantageous in both productivity and cell line stability. Binding affinity and N-glycan analysis of the antibody revealed that all batches of product were of similar quality independent on integrated sites. Deep sequencing demonstrated that there was low level of off-target mutations caused by CRISPR/Cas9, but none of them contributed to the development process of transgene cell lines. Our results demonstrated the feasibility of C12orf35 as the target site for exogenous gene integration, and strongly suggested that C12orf35 targeted integration mediated by CRISPR/Cas9 is a reliable strategy for the rapid development of recombinant CHO cell lines.

Bacteriophage-based Vectors for Site-specific Insertion of DNA in the Chromosome of Corynebacteria

PubMed Central

Oram, Mark; Woolston, Joelle E.; Jacobson, Andrew D.; Holmes, Randall K.; Oram, Diana M.

2007-01-01

In Corynebacterium diphtheriae, diphtheria toxin is encoded by the tox gene of some temperate corynephages such as β. β-like corynephages are capable of inserting into the C. diphtheriae chromosome at two specific sites, attB1 and attB2. Transcription of the phage-encoded tox gene, and many chromosomally-encoded genes, is regulated by the DtxR protein in response to Fe2+ levels. Characterizing DtxR-dependent gene regulation is pivotal in understanding diphtheria pathogenesis and mechanisms of iron-dependent gene expression; although this has been hampered by a lack of molecular genetic tools in C. diphtheriae and related Coryneform species. To expand the systems for genetic manipulation of C. diphtheriae, we constructed plasmid vectors capable of integrating into the chromosome. These plasmids contain the β-encoded attP site and the DIP0182 integrase gene of C. diphtheriae NCTC13129. When these vectors were delivered to the cytoplasm of non-lysogenic C. diphtheriae, they integrated into either the attB1 or attB2 sites with comparable frequency. Lysogens were also transformed with these vectors, by virtue of the second attB site. An integrated vector carrying an intact dtxR gene complemented the mutant phenotypes of a C. diphtheriae ΔdtxR strain. Additionally, strains of β-susceptible C. ulcerans, and C. glutamicum, a species non-permissive for β, were each transformed with these vectors. This work significantly extends the tools available for targeted transformation of both pathogenic and non-pathogenic Corynebacterium species. PMID:17275217

Advancedmonitoring Systems for Landslide Risk Reduction in THE'SIQ' of PETRA (jordan)

NASA Astrophysics Data System (ADS)

Delmonaco, G.; Brini, M.; Cesaro, G.

2017-08-01

The UNESCO World Heritage Site of Petra is characterized by a spectacular geo-archaeological landscape that lead to its inscription as World Heritage Site in 1985. Petra is also a fragile site facing a wide diversity of risks, ranging from those posed by environmental factors as well as those attributed to tourism. In recent years, hazardous natural phenomena were registered as increasingly impacting the site, and most specifically the 'Siq', a 1.2 km naturally formed gorge serving as the only tourist entrance to the archaeological park, posing a major threat to cultural heritage and visitors. These recent events have prompted UNESCO Amman Office, in cooperation with the national authorities, to develop a strategy towards prevention and mitigation of instability phenomena at the 'Siq' and, thus, further contribute to the management and conservation of the site through the implementation of the multi-year Italian funded "Siq Stability" project Actions have been primarily focusing on the analysis of the stability conditions of the 'Siq' slopes, the installation of an integrated monitoring system and the definition and implementation of mitigation measures against rock instability. This paper reports a detailed description of the integrated monitoring system installed paying particular attention on the wireless monitoring devices and the EASA applications, which proved to be some of the most successful systems implemented in the framework of the project. Some preliminary results regarding the data retrieved, policies applied and actions taken to ensure long-term sustainability and capacity development of the national authorities are also reported.

Using two classification schemes to develop vegetation indices of biological integrity for wetlands in West Virginia, USA.

PubMed

Veselka, Walter; Rentch, James S; Grafton, William N; Kordek, Walter S; Anderson, James T

2010-11-01

Bioassessment methods for wetlands, and other bodies of water, have been developed worldwide to measure and quantify changes in "biological integrity." These assessments are based on a classification system, meant to ensure appropriate comparisons between wetland types. Using a local site-specific disturbance gradient, we built vegetation indices of biological integrity (Veg-IBIs) based on two commonly used wetland classification systems in the USA: One based on vegetative structure and the other based on a wetland's position in a landscape and sources of water. The resulting class-specific Veg-IBIs were comprised of 1-5 metrics that varied in their sensitivity to the disturbance gradient (R2=0.14-0.65). Moreover, the sensitivity to the disturbance gradient increased as metrics from each of the two classification schemes were combined (added). Using this information to monitor natural and created wetlands will help natural resource managers track changes in biological integrity of wetlands in response to anthropogenic disturbance and allows the use of vegetative communities to set ecological performance standards for mitigation banks.

Evaluation of the Enhanced Integrated Climatic Model for modulus-based construction specification for Oklahoma pavements.

DOT National Transportation Integrated Search

2013-07-01

The study provides estimation of site specific variation in environmental factors that can be : used in predicting seasonal and long-term variations in moduli of unbound materials. Using : these site specific estimates, the EICM climatic input files ...

CEO Sites Mission Management System (SMMS)

NASA Technical Reports Server (NTRS)

Trenchard, Mike

2014-01-01

Late in fiscal year 2011, the Crew Earth Observations (CEO) team was tasked to upgrade its science site database management tool, which at the time was integrated with the Automated Mission Planning System (AMPS) originally developed for Earth Observations mission planning in the 1980s. Although AMPS had been adapted and was reliably used by CEO for International Space Station (ISS) payload operations support, the database structure was dated, and the compiler required for modifications would not be supported in the Windows 7 64-bit operating system scheduled for implementation the following year. The Sites Mission Management System (SMMS) is now the tool used by CEO to manage a heritage Structured Query Language (SQL) database of more than 2,000 records for Earth science sites. SMMS is a carefully designed and crafted in-house software package with complete and detailed help files available for the user and meticulous internal documentation for future modifications. It was delivered in February 2012 for test and evaluation. Following acceptance, it was implemented for CEO mission operations support in April 2012. The database spans the period from the earliest systematic requests for astronaut photography during the shuttle era to current ISS mission support of the CEO science payload. Besides logging basic image information (site names, locations, broad application categories, and mission requests), the upgraded database management tool now tracks dates of creation, modification, and activation; imagery acquired in response to requests; the status and location of ancillary site information; and affiliations with studies, their sponsors, and collaborators. SMMS was designed to facilitate overall mission planning in terms of site selection and activation and provide the necessary site parameters for the Satellite Tool Kit (STK) Integrated Message Production List Editor (SIMPLE), which is used by CEO operations to perform daily ISS mission planning. The CEO team uses the SMMS for three general functions - database queries of content and status, individual site creation and updates, and mission planning. The CEO administrator of the science site database is able to create or modify the content of sites and activate or deactivate them based on the requirements of the sponsors. The administrator supports and implements ISS mission planning by assembling, reporting, and activating mission-specific site selections for management; deactivating sites as requirements are met; and creating new sites, such as International Charter sites for disasters, as circumstances warrant. In addition to the above CEO internal uses, when site planning for a specific ISS mission is complete and approved, the SMMS can produce and export those essential site database elements for the mission into XML format for use by onboard Earth-location systems, such as Worldmap. The design, development, and implementation of the SMMS resulted in a superior database management system for CEO science sites by focusing on the functions and applications of the database alone instead of integrating the database with the multipurpose configuration of the AMPS. Unlike the AMPS, it can function and be modified within the existing Windows 7 environment. The functions and applications of the SMMS were expanded to accommodate more database elements, report products, and a streamlined interface for data entry and review. A particularly elegant enhancement in data entry was the integration of the Google Earth application for the visual display and definition of site coordinates for site areas defined by multiple coordinates. Transfer between the SMMS and Google Earth is accomplished with a Keyhole Markup Language (KML) expression of geographic data (see figures 3 and 4). Site coordinates may be entered into the SMMS panel directly for display in Google Earth, or the coordinates may be defined on the Google Earth display as a mouse-controlled polygonal definition and transferred back into the SMMS as KML input. This significantly reduces the possibility of errors in coordinate entries and provides visualization of the scale of the site being defined. CEO now has a powerful tool for managing and defining sites on the Earth's surface for both targets of astronaut photography or other onboard remote sensing systems. It can also record and track results by sponsor, collaborator, or type of study.

Combining Modeling and Monitoring to Produce a New Paradigm of an Integrated Approach to Providing Long-Term Control of Contaminants

NASA Astrophysics Data System (ADS)

Fogwell, T. W.

2009-12-01

Sir David King, Chief Science Advisor to the British government and Cambridge University Professor, stated in October 2005, "The scientific community is considerably more capable than it has been in the past to assist governments to avoid and reduce risk to their own populations. Prime ministers and presidents ignore the advice from the science community at the peril of their own populations." Some of these greater capabilities can be found in better monitoring techniques applied to better modeling methods. These modeling methods can be combined with the information derived from monitoring data in order to decrease the risk of population exposure to dangerous substances and to promote efficient control or cleanup of the contaminants. An introduction is presented of the types of problems that exist for long-term control of radionuclides at DOE sites. A breakdown of the distributions at specific sites is given, together with the associated difficulties. A paradigm for remediation showing the integration of monitoring with modeling is presented. It is based on a feedback system that allows for the monitoring to act as principal sensors in a control system. The resulting system can be optimized to improve performance. Optimizing monitoring automatically entails linking the monitoring with modeling. If monitoring designs were required to be more efficient, thus requiring optimization, then the monitoring automatically becomes linked to modeling. Records of decision could be written to accommodate revisions in monitoring as better modeling evolves. Currently the establishment of a very prescriptive monitoring program fails to have a mechanism for improving models and improving control of the contaminants. The technical pieces of the required paradigm are already available; they just need to be implemented and applied to solve the long-term control of the contaminants. An integration of the various parts of the system is presented. Each part is described, and examples are given. References are given to other projects which bring together similar elements in systems for the control of contaminants. Trends are given for the development of the technical features of a robust system. Examples of monitoring methods for specific sites are given. The examples are used to illustrate how such a system would work. Examples of technology needs are presented. Finally, other examples of integrated modeling-monitoring approaches are presented.

Structural Insights into the Assembly of the Adeno-associated Virus Type 2 Rep68 Protein on the Integration Site AAVS1*

PubMed Central

Musayev, Faik N.; Zarate-Perez, Francisco; Bishop, Clayton; Burgner, John W.; Escalante, Carlos R.

2015-01-01

Adeno-associated virus (AAV) is the only eukaryotic virus with the property of establishing latency by integrating site-specifically into the human genome. The integration site known as AAVS1 is located in chromosome 19 and contains multiple GCTC repeats that are recognized by the AAV non-structural Rep proteins. These proteins are multifunctional, with an N-terminal origin-binding domain (OBD) and a helicase domain joined together by a short linker. As a first step to understand the process of site-specific integration, we proceeded to characterize the recognition and assembly of Rep68 onto the AAVS1 site. We first determined the x-ray structure of AAV-2 Rep68 OBD in complex with the AAVS1 DNA site. Specificity is achieved through the interaction of a glycine-rich loop that binds the major groove and an α-helix that interacts with a downstream minor groove on the same face of the DNA. Although the structure shows a complex with three OBD molecules bound to the AAVS1 site, we show by using analytical centrifugation and electron microscopy that the full-length Rep68 forms a heptameric complex. Moreover, we determined that a minimum of two direct repeats is required to form a stable complex and to melt DNA. Finally, we show that although the individual domains bind DNA poorly, complex assembly requires oligomerization and cooperation between its OBD, helicase, and the linker domains. PMID:26370092

Location of the unique integration site on an Escherichia coli chromosome by bacteriophage lambda DNA in vivo.

PubMed

Tal, Asaf; Arbel-Goren, Rinat; Costantino, Nina; Court, Donald L; Stavans, Joel

2014-05-20

The search for specific sequences on long genomes is a key process in many biological contexts. How can specific target sequences be located with high efficiency, within physiologically relevant times? We addressed this question for viral integration, a fundamental mechanism of horizontal gene transfer driving prokaryotic evolution, using the infection of Escherichia coli bacteria with bacteriophage λ and following the establishment of a lysogenic state. Following the targeting process in individual live E. coli cells in real time revealed that λ DNA remains confined near the entry point of a cell following infection. The encounter between the 15-bp-long target sequence on the chromosome and the recombination site on the viral genome is facilitated by the directed motion of bacterial DNA generated during chromosome replication, in conjunction with constrained diffusion of phage DNA. Moving the native bacterial integration site to different locations on the genome and measuring the integration frequency in these strains reveals that the frequencies of the native site and a site symmetric to it relative to the origin are similar, whereas both are significantly higher than when the integration site is moved near the terminus, consistent with the replication-driven mechanism we propose. This novel search mechanism is yet another example of the exquisite coevolution of λ with its host.

Characterization of the temperate phage vB_RleM_PPF1 and its site-specific integration into the Rhizobium leguminosarum F1 genome.

PubMed

Halmillawewa, Anupama P; Restrepo-Córdoba, Marcela; Perry, Benjamin J; Yost, Christopher K; Hynes, Michael F

2016-02-01

Bacteriophages may play an important role in regulating population size and diversity of the root nodule symbiont Rhizobium leguminosarum, as well as participating in horizontal gene transfer. Although phages that infect this species have been isolated in the past, our knowledge of their molecular biology, and especially of genome composition, is extremely limited, and this lack of information impacts on the ability to assess phage population dynamics and limits potential agricultural applications of rhizobiophages. To help address this deficit in available sequence and biological information, the complete genome sequence of the Myoviridae temperate phage PPF1 that infects R. leguminosarum biovar viciae strain F1 was determined. The genome is 54,506 bp in length with an average G+C content of 61.9 %. The genome contains 94 putative open reading frames (ORFs) and 74.5 % of these predicted ORFs share homology at the protein level with previously reported sequences in the database. However, putative functions could only be assigned to 25.5 % (24 ORFs) of the predicted genes. PPF1 was capable of efficiently lysogenizing its rhizobial host R. leguminosarum F1. The site-specific recombination system of the phage targets an integration site that lies within a putative tRNA-Pro (CGG) gene in R. leguminosarum F1. Upon integration, the phage is capable of restoring the disrupted tRNA gene, owing to the 50 bp homologous sequence (att core region) it shares with its rhizobial host genome. Phage PPF1 is the first temperate phage infecting members of the genus Rhizobium for which a complete genome sequence, as well as other biological data such as the integration site, is available.

3D simulation as a tool for improving the safety culture during remediation work at Andreeva Bay.

PubMed

Chizhov, K; Sneve, M K; Szőke, I; Mazur, I; Mark, N K; Kudrin, I; Shandala, N; Simakov, A; Smith, G M; Krasnoschekov, A; Kosnikov, A; Kemsky, I; Kryuchkov, V

2014-12-01

Andreeva Bay in northwest Russia hosts one of the former coastal technical bases of the Northern Fleet. Currently, this base is designated as the Andreeva Bay branch of Northwest Center for Radioactive Waste Management (SevRAO) and is a site of temporary storage (STS) for spent nuclear fuel (SNF) and other radiological waste generated during the operation and decommissioning of nuclear submarines and ships. According to an integrated expert evaluation, this site is the most dangerous nuclear facility in northwest Russia. Environmental rehabilitation of the site is currently in progress and is supported by strong international collaboration. This paper describes how the optimization principle (ALARA) has been adopted during the planning of remediation work at the Andreeva Bay STS and how Russian-Norwegian collaboration greatly contributed to ensuring the development and maintenance of a high level safety culture during this process. More specifically, this paper describes how integration of a system, specifically designed for improving the radiological safety of workers during the remediation work at Andreeva Bay, was developed in Russia. It also outlines the 3D radiological simulation and virtual reality based systems developed in Norway that have greatly facilitated effective implementation of the ALARA principle, through supporting radiological characterisation, work planning and optimization, decision making, communication between teams and with the authorities and training of field operators.

Fitness cost implications of phiC31-mediated site-specific integrations in target-site strains of the Mexican fruit fly, Anastrepha ludens (Diptera: Tephritidae)

USDA-ARS?s Scientific Manuscript database

Site-specific recombination technologies are powerful new tools for the manipulation of genomic DNA in insects that can improve transgenesis strategies such as targeting transgene insertions, allowing transgene cassette exchange and DNA mobilization for transgene stabilization. However, understandin...

Integrated genetic and epigenetic prediction of coronary heart disease in the Framingham Heart Study.

PubMed

Dogan, Meeshanthini V; Grumbach, Isabella M; Michaelson, Jacob J; Philibert, Robert A

2018-01-01

An improved method for detecting coronary heart disease (CHD) could have substantial clinical impact. Building on the idea that systemic effects of CHD risk factors are a conglomeration of genetic and environmental factors, we use machine learning techniques and integrate genetic, epigenetic and phenotype data from the Framingham Heart Study to build and test a Random Forest classification model for symptomatic CHD. Our classifier was trained on n = 1,545 individuals and consisted of four DNA methylation sites, two SNPs, age and gender. The methylation sites and SNPs were selected during the training phase. The final trained model was then tested on n = 142 individuals. The test data comprised of individuals removed based on relatedness to those in the training dataset. This integrated classifier was capable of classifying symptomatic CHD status of those in the test set with an accuracy, sensitivity and specificity of 78%, 0.75 and 0.80, respectively. In contrast, a model using only conventional CHD risk factors as predictors had an accuracy and sensitivity of only 65% and 0.42, respectively, but with a specificity of 0.89 in the test set. Regression analyses of the methylation signatures illustrate our ability to map these signatures to known risk factors in CHD pathogenesis. These results demonstrate the capability of an integrated approach to effectively model symptomatic CHD status. These results also suggest that future studies of biomaterial collected from longitudinally informative cohorts that are specifically characterized for cardiac disease at follow-up could lead to the introduction of sensitive, readily employable integrated genetic-epigenetic algorithms for predicting onset of future symptomatic CHD.

Accuracy and efficiency define Bxb1 integrase as the best of fifteen candidate serine recombinases for the integration of DNA into the human genome

PubMed Central

2013-01-01

Background Phage-encoded serine integrases, such as φC31 integrase, are widely used for genome engineering. Fifteen such integrases have been described but their utility for genome engineering has not been compared in uniform assays. Results We have compared fifteen serine integrases for their utility for DNA manipulations in mammalian cells after first demonstrating that all were functional in E. coli. Chromosomal recombination reporters were used to show that seven integrases were active on chromosomally integrated DNA in human fibroblasts and mouse embryonic stem cells. Five of the remaining eight enzymes were active on extra-chromosomal substrates thereby demonstrating that the ability to mediate extra-chromosomal recombination is no guide to ability to mediate site-specific recombination on integrated DNA. All the integrases that were active on integrated DNA also promoted DNA integration reactions that were not mediated through conservative site-specific recombination or damaged the recombination sites but the extent of these aberrant reactions varied over at least an order of magnitude. Bxb1 integrase yielded approximately two-fold more recombinants and displayed about two fold less damage to the recombination sites than the next best recombinase; φC31 integrase. Conclusions We conclude that the Bxb1 and φC31 integrases are the reagents of choice for genome engineering in vertebrate cells and that DNA damage repair is a major limitation upon the utility of this class of site-specific recombinase. PMID:24139482

Site directed recombination

DOEpatents

Jurka, Jerzy W.

1997-01-01

Enhanced homologous recombination is obtained by employing a consensus sequence which has been found to be associated with integration of repeat sequences, such as Alu and ID. The consensus sequence or sequence having a single transition mutation determines one site of a double break which allows for high efficiency of integration at the site. By introducing single or double stranded DNA having the consensus sequence flanking region joined to a sequence of interest, one can reproducibly direct integration of the sequence of interest at one or a limited number of sites. In this way, specific sites can be identified and homologous recombination achieved at the site by employing a second flanking sequence associated with a sequence proximal to the 3'-nick.

Techniques to Access Databases and Integrate Data for Hydrologic Modeling

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

Whelan, Gene; Tenney, Nathan D.; Pelton, Mitchell A.

2009-06-17

This document addresses techniques to access and integrate data for defining site-specific conditions and behaviors associated with ground-water and surface-water radionuclide transport applicable to U.S. Nuclear Regulatory Commission reviews. Environmental models typically require input data from multiple internal and external sources that may include, but are not limited to, stream and rainfall gage data, meteorological data, hydrogeological data, habitat data, and biological data. These data may be retrieved from a variety of organizations (e.g., federal, state, and regional) and source types (e.g., HTTP, FTP, and databases). Available data sources relevant to hydrologic analyses for reactor licensing are identified and reviewed.more » The data sources described can be useful to define model inputs and parameters, including site features (e.g., watershed boundaries, stream locations, reservoirs, site topography), site properties (e.g., surface conditions, subsurface hydraulic properties, water quality), and site boundary conditions, input forcings, and extreme events (e.g., stream discharge, lake levels, precipitation, recharge, flood and drought characteristics). Available software tools for accessing established databases, retrieving the data, and integrating it with models were identified and reviewed. The emphasis in this review was on existing software products with minimal required modifications to enable their use with the FRAMES modeling framework. The ability of four of these tools to access and retrieve the identified data sources was reviewed. These four software tools were the Hydrologic Data Acquisition and Processing System (HDAPS), Integrated Water Resources Modeling System (IWRMS) External Data Harvester, Data for Environmental Modeling Environmental Data Download Tool (D4EM EDDT), and the FRAMES Internet Database Tools. The IWRMS External Data Harvester and the D4EM EDDT were identified as the most promising tools based on their ability to access and retrieve the required data, and their ability to integrate the data into environmental models using the FRAMES environment.« less

INVESTIGATING UNCERTAINTY AND SENSITIVITY IN INTEGRATED, MULTIMEDIA ENVIRONMENTAL MODELS: TOOLS FOR FRAMES-3MRA

EPA Science Inventory

Elucidating uncertainty and sensitivity structures in environmental models can be a difficult task, even for low-order, single-medium constructs driven by a unique set of site-specific data. Quantitative assessment of integrated, multimedia models that simulate hundreds of sites...

A Team-Based Process for Designing Comprehensive, Integrated, Three-Tiered (CI3T) Models of Prevention: How Does My School-Site Leadership Team Design a CI3T Model?

ERIC Educational Resources Information Center

Lane, Kathleen Lynne; Oakes, Wendy Peia; Jenkins, Abbie; Menzies, Holly Mariah; Kalberg, Jemma Robertson

2014-01-01

Comprehensive, integrated, three-tiered models are context specific and developed by school-site teams according to the core values held by the school community. In this article, the authors provide a step-by-step, team-based process for designing comprehensive, integrated, three-tiered models of prevention that integrate academic, behavioral, and…

Random integration of SV40 in SV40-transformed, immortalized human fibroblasts.

PubMed

Hara, H; Kaji, H

1987-02-01

We have studied the relationship between immortalization of SV40-transformed human embryonic fibroblasts and their SV40 integration sites. From several independently transformed cell pools, we have isolated clones which do not harbor unintegrated SV40 DNA. We have analysed whole-cell DNA from these clones, using the Southern blot method. Our results suggest that no specific integration sites in the cellular genome exist which are a prerequisite for the immortalization process. Although some integration sites were found to be predominant in pre-crisis clones, they could not be detected in the post-crisis clones. This suggests that none of these predominating sites is selected for during the crisis period.

Characterization of the genetic elements required for site-specific integration of plasmid pSE211 in Saccharopolyspora erythraea.

PubMed Central

Brown, D P; Idler, K B; Katz, L

1990-01-01

The 18.1-kilobase plasmid pSE211 integrates into the chromosome of Saccharopolyspora erythraea at a specific attB site. Restriction analysis of the integrated plasmid, pSE211int, and adjacent chromosomal sequences allowed identification of attP, the plasmid attachment site. Nucleotide sequencing of attP, attB, attL, and attR revealed a 57-base-pair sequence common to all sites with no duplications of adjacent plasmid or chromosomal sequences in the integrated state, indicating that integration takes place through conservative, reciprocal strand exchange. An analysis of the sequences indicated the presence of a putative gene for Phe-tRNA at attB which is preserved at attL after integration has occurred. A comparison of the attB site for a number of actinomycete plasmids is presented. Integration at attB was also observed when a 2.4-kilobase segment of pSE211 containing attP and the adjacent plasmid sequence was used to transform a pSE211- host. Nucleotide sequencing of this segment revealed the presence of two complete open reading frames (ORFs) and a segment of a third ORF. The ORF adjacent to attP encodes a putative polypeptide 437 amino acids in length that shows similarity, at its C-terminal domain, to sequences of site-specific recombinases of the integrase family. The adjacent ORF encodes a putative 98-amino-acid basic polypeptide that contains a helix-turn-helix motif at its N terminus which corresponds to domains in the Xis proteins of a number of bacteriophages. A proposal for the function of this polypeptide is presented. The deduced amino acid sequence of the third ORF did not reveal similarities to polypeptide sequences in the current data banks. Images FIG. 2 FIG. 3 PMID:2180909

Mitigation of nutrient losses via surface runoff from rice cropping systems with alternate wetting and drying irrigation and site-specific nutrient management practices.

PubMed

Liang, X Q; Chen, Y X; Nie, Z Y; Ye, Y S; Liu, J; Tian, G M; Wang, G H; Tuong, T P

2013-10-01

Resource-conserving irrigation and fertilizer management practices have been developed for rice systems which may help address water quality concerns by reducing N and P losses via surface runoff. Field experiments under three treatments, i.e., farmers' conventional practice (FCP), alternate wetting and drying (AWD), and AWD integrated with site-specific nutrient management (AWD + SSNM) were carried out during two rice seasons at two sites in the southwest Yangtze River delta region. Across site years, results indicated that under AWD irrigation (i.e., AWD and AWD + SSNM), water inputs were reduced by 13.4~27.5 % and surface runoff was reduced by 30.2~36.7 % compared to FCP. When AWD was implemented alone, total N and P loss masses via surface runoff were reduced by 23.3~30.4 % and 26.9~31.7 %, respectively, compared to FCP. However, nutrient concentrations of surface runoff did not decrease under AWD alone. Under AWD + SSNM, total N and P loss masses via surface runoff were reduced to a greater extent than AWD alone (39.4~47.6 % and 46.1~48.3 % compared to FCP, respectively), while fertilizer inputs and N surpluses significantly decreased and rice grain yields increased relative to FCP. Therefore, by more closely matching nutrient supply with crop demand and reducing both surface runoff and nutrient concentrations of surface runoff, our results demonstrate that integration of AWD and SSNM practices can mitigate N and P losses via surface runoff from rice fields while maintaining high yields.

Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species

PubMed Central

Tlapák, Hana; Köppen, Kristin; Rydzewski, Kerstin; Grunow, Roland; Heuner, Klaus

2018-01-01

We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of Francisella hispaniensis AS02-814 (F. tularensis subsp. novicida-like 3523). In this study, we constructed two variants of a Francisella phage integration vector, called pFIV1-Val and pFIV2-Val (Francisella Integration Vector-tRNAVal-specific), using the attL/R-sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a sacB gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in Escherichia coli. The constructs generated a circular episomal form in E. coli which could be used to transform Francisella spp. where FIV-Val stably integrated site specifically into the tRNAVal gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a Francisella mutant strain. The vectors were stable in vitro and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in Francisella research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different Francisella species. PMID:29594068

Construction of a New Phage Integration Vector pFIV-Val for Use in Different Francisella Species.

PubMed

Tlapák, Hana; Köppen, Kristin; Rydzewski, Kerstin; Grunow, Roland; Heuner, Klaus

2018-01-01

We recently identified and described a putative prophage on the genomic island FhaGI-1 located within the genome of Francisella hispaniensis AS02-814 ( F. tularensis subsp. novicida -like 3523). In this study, we constructed two variants of a Francisella phage integration vector, called pFIV1-Val and pFIV2-Val ( Francisella Integration Vector-tRNA Val -specific), using the attL/R- sites and the site-specific integrase (FN3523_1033) of FhaGI-1, a chloramphenicol resistance cassette and a sacB gene for counter selection of transformants against the vector backbone. We inserted the respective sites and genes into vector pUC57-Kana to allow for propagation in Escherichia coli . The constructs generated a circular episomal form in E. coli which could be used to transform Francisella spp . where FIV-Val stably integrated site specifically into the tRNA Val gene of the genome, whereas pUC57-Kana is lost due to counter selection. Functionality of the new vector was demonstrated by the successfully complementation of a Francisella mutant strain. The vectors were stable in vitro and during host-cell infection without selective pressure. Thus, the vectors can be applied as a further genetic tool in Francisella research, expanding the present genetic tools by an integrative element. This new element is suitable to perform long-term experiments with different Francisella species.

Determinants for DNA target structure selectivity of the human LINE-1 retrotransposon endonuclease.

PubMed

Repanas, Kostas; Zingler, Nora; Layer, Liliana E; Schumann, Gerald G; Perrakis, Anastassis; Weichenrieder, Oliver

2007-01-01

The human LINE-1 endonuclease (L1-EN) is the targeting endonuclease encoded by the human LINE-1 (L1) retrotransposon. L1-EN guides the genomic integration of new L1 and Alu elements that presently account for approximately 28% of the human genome. L1-EN bears considerable technological interest, because its target selectivity may ultimately be engineered to allow the site-specific integration of DNA into defined genomic locations. Based on the crystal structure, we generated L1-EN mutants to analyze and manipulate DNA target site recognition. Crystal structures and their dynamic and functional analysis show entire loop grafts to be feasible, resulting in altered specificity, while individual point mutations do not change the nicking pattern of L1-EN. Structural parameters of the DNA target seem more important for recognition than the nucleotide sequence, and nicking profiles on DNA oligonucleotides in vitro are less well defined than the respective integration site consensus in vivo. This suggests that additional factors other than the DNA nicking specificity of L1-EN contribute to the targeted integration of non-LTR retrotransposons.

Managing health and safety risks: Implications for tailoring health and safety management system practices.

PubMed

Willmer, D R; Haas, E J

2016-01-01

As national and international health and safety management system (HSMS) standards are voluntarily accepted or regulated into practice, organizations are making an effort to modify and integrate strategic elements of a connected management system into their daily risk management practices. In high-risk industries such as mining, that effort takes on added importance. The mining industry has long recognized the importance of a more integrated approach to recognizing and responding to site-specific risks, encouraging the adoption of a risk-based management framework. Recently, the U.S. National Mining Association led the development of an industry-specific HSMS built on the strategic frameworks of ANSI: Z10, OHSAS 18001, The American Chemistry Council's Responsible Care, and ILO-OSH 2001. All of these standards provide strategic guidance and focus on how to incorporate a plan-do-check-act cycle into the identification, management and evaluation of worksite risks. This paper details an exploratory study into whether practices associated with executing a risk-based management framework are visible through the actions of an organization's site-level management of health and safety risks. The results of this study show ways that site-level leaders manage day-to-day risk at their operations that can be characterized according to practices associated with a risk-based management framework. Having tangible operational examples of day-to-day risk management can serve as a starting point for evaluating field-level risk assessment efforts and their alignment to overall company efforts at effective risk mitigation through a HSMS or other processes.

Remedial action assessment system: Decision support for environmental cleanup

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

Pennock, K.A.; Bohn, S.; Franklin, A.L.

1991-11-01

A large number of hazardous waste sites across the United States await treatment. Waste sites can be physically complex entities composed of multiple, possibly interacting contaminants distributed throughout one or more media. The sites may be active as well with contaminants escaping through one or more potential escape paths. Treatment of these sites requires a long and costly commitment involving the coordination of activities among several waste treatment professionals. In order to reduce the cost and time required for the specification of treatment at these waste sites. The Remedial Action Assessment System (RAAS) was proposed. RAAS is an automated informationmore » management system which utilizes a combination of expert reasoning and numerical models to produce the combinations of treatment technologies, known as treatment trains, which satisfy the treatment objectives of a particular site. In addition, RAAS supports the analysis of these trains with regard to effectiveness and cost so that the viable treatment trains can be measured against each other. The Remedial Action Assessment System is a hybrid system designed and constructed using object-oriented tools and techniques. RAAS is advertised as a hybrid system because it combines, in integral fashion, numerical computing (primarily quantitative models) with expert system reasoning. An object-oriented approach was selected due to many of its inherent advantages, among these the naturalness of modeling physical objects and processes.« less

Wind Power Opportunities in St. Thomas, USVI: A Site-Specific Evaluation and Analysis

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

Lantz, E.; Warren, A.; Roberts, J. O.

This NREL technical report utilizes a development framework originated by NREL and known by the acronym SROPTTC to assist the U.S. Virgin Islands in identifying and understanding concrete opportunities for wind power development in the territory. The report covers each of the seven components of the SROPTTC framework: Site, Resource, Off-take, Permitting, Technology, Team, and Capital as they apply to wind power in the USVI and specifically to a site in Bovoni, St. Thomas. The report concludes that Bovoni peninsula is a strong candidate for utility-scale wind generation in the territory. It represents a reasonable compromise in terms of windmore » resource, distance from residences, and developable terrain. Hurricane risk and variable terrain on the peninsula and on potential equipment transport routes add technical and logistical challenges but do not appear to represent insurmountable barriers. In addition, integration of wind power into the St. Thomas power system will present operational challenges, but based on experience in other islanded power systems, there are reasonable solutions for addressing these challenges.« less

DNA minicircles clarify the specific role of DNA structure on retroviral integration

PubMed Central

Pasi, Marco; Mornico, Damien; Volant, Stevenn; Juchet, Anna; Batisse, Julien; Bouchier, Christiane; Parissi, Vincent; Ruff, Marc; Lavery, Richard; Lavigne, Marc

2016-01-01

Chromatin regulates the selectivity of retroviral integration into the genome of infected cells. At the nucleosome level, both histones and DNA structure are involved in this regulation. We propose a strategy that allows to specifically study a single factor: the DNA distortion induced by the nucleosome. This strategy relies on mimicking this distortion using DNA minicircles (MCs) having a fixed rotational orientation of DNA curvature, coupled with atomic-resolution modeling. Contrasting MCs with linear DNA fragments having identical sequences enabled us to analyze the impact of DNA distortion on the efficiency and selectivity of integration. We observed a global enhancement of HIV-1 integration in MCs and an enrichment of integration sites in the outward-facing DNA major grooves. Both of these changes are favored by LEDGF/p75, revealing a new, histone-independent role of this integration cofactor. PFV integration is also enhanced in MCs, but is not associated with a periodic redistribution of integration sites, thus highlighting its distinct catalytic properties. MCs help to separate the roles of target DNA structure, histone modifications and integrase (IN) cofactors during retroviral integration and to reveal IN-specific regulation mechanisms. PMID:27439712

A Geographical Information System Based Approach for Integrated Strategies of Tick Surveillance and Control in the Peri-Urban Natural Reserve of Monte Pellegrino (Palermo, Southern Italy).

PubMed

Torina, Alessandra; Blanda, Valeria; Blanda, Marcellocalogero; Auteri, Michelangelo; La Russa, Francesco; Scimeca, Salvatore; D'Agostino, Rosalia; Disclafani, Rosaria; Villari, Sara; Currò, Vittoria; Caracappa, Santo

2018-02-27

Ticks (Acari: Ixodidae) are bloodsucking arthropods involved in pathogen transmission in animals and humans. Tick activity depends on various ecological factors such as vegetation, hosts, and temperature. The aim of this study was to analyse the spatial/temporal distribution of ticks in six sites within a peri-urban area of Palermo (Natural Reserve of Monte Pellegrino) and correlate it with field data using Geographical Information System (GIS) data. A total of 3092 ticks were gathered via dragging method from June 2012 to May 2014. The species collected were: Ixodes ventalloi (46.09%), Hyalomma lusitanicum (19.99%), Rhipicephalus sanguineus (17.34%), Rhipicephalus pusillus (16.11%), Haemaphisalis sulcata (0.36%), Dermacentor marginatus (0.10%), and Rhipicephalus turanicus (0.03%). GIS analysis revealed environmental characteristics of each site, and abundance of each tick species was analysed in relation to time (monthly trend) and space (site-specific abundance). A relevant presence of I. ventalloi in site 2 and H. lusitanicum in site 5 was observed, suggesting the possible exposure of animals and humans to tick-borne pathogens. Our study shows the importance of surveillance of ticks in peri-urban areas and the useful implementation of GIS analysis in vector ecology; studies on temporal and spatial distribution of ticks correlated to GIS-based ecological analysis represent an integrated strategy for decision support in public health.

DbPTM 3.0: an informative resource for investigating substrate site specificity and functional association of protein post-translational modifications.

PubMed

Lu, Cheng-Tsung; Huang, Kai-Yao; Su, Min-Gang; Lee, Tzong-Yi; Bretaña, Neil Arvin; Chang, Wen-Chi; Chen, Yi-Ju; Chen, Yu-Ju; Huang, Hsien-Da

2013-01-01

Protein modification is an extremely important post-translational regulation that adjusts the physical and chemical properties, conformation, stability and activity of a protein; thus altering protein function. Due to the high throughput of mass spectrometry (MS)-based methods in identifying site-specific post-translational modifications (PTMs), dbPTM (http://dbPTM.mbc.nctu.edu.tw/) is updated to integrate experimental PTMs obtained from public resources as well as manually curated MS/MS peptides associated with PTMs from research articles. Version 3.0 of dbPTM aims to be an informative resource for investigating the substrate specificity of PTM sites and functional association of PTMs between substrates and their interacting proteins. In order to investigate the substrate specificity for modification sites, a newly developed statistical method has been applied to identify the significant substrate motifs for each type of PTMs containing sufficient experimental data. According to the data statistics in dbPTM, >60% of PTM sites are located in the functional domains of proteins. It is known that most PTMs can create binding sites for specific protein-interaction domains that work together for cellular function. Thus, this update integrates protein-protein interaction and domain-domain interaction to determine the functional association of PTM sites located in protein-interacting domains. Additionally, the information of structural topologies on transmembrane (TM) proteins is integrated in dbPTM in order to delineate the structural correlation between the reported PTM sites and TM topologies. To facilitate the investigation of PTMs on TM proteins, the PTM substrate sites and the structural topology are graphically represented. Also, literature information related to PTMs, orthologous conservations and substrate motifs of PTMs are also provided in the resource. Finally, this version features an improved web interface to facilitate convenient access to the resource.

An evaluation of contaminated estuarine sites using sediment quality guidelines and ecological assessment methodologies.

PubMed

Fulton, M; Key, P; Wirth, E; Leight, A K; Daugomah, J; Bearden, D; Sivertsen, S; Scott, G

2006-10-01

Toxic contaminants may enter estuarine ecosystems through a variety of pathways. When sediment contaminant levels become sufficiently high, they may impact resident biota. One approach to predict sediment-associated toxicity in estuarine ecosystems involves the use of sediment quality guidelines (ERMs, ERLs) and site-specific contaminant chemistry while a second approach utilizes site-specific ecological sampling to assess impacts at the population or community level. The goal of this study was to utilize an integrated approach including chemical contaminant analysis, sediment quality guidelines and grass shrimp population monitoring to evaluate the impact of contaminants from industrial sources. Three impacted sites and one reference site were selected for study. Grass shrimp populations were sampled using a push-netting approach. Sediment samples were collected at each site and analyzed for metals, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and pesticides. Contaminant levels were then compared to sediment quality guidelines. In general, grass shrimp population densities at the sites decreased as the ERM quotients increased. Grass shrimp densities were significantly reduced at the impacted site that had an ERM exceedance for chromium and the highest Mean ERM quotient. Regression analysis indicated that sediment chromium concentrations were negatively correlated with grass shrimp density. Grass shrimp size was reduced at two sites with intermediate levels of contamination. These findings support the use of both sediment quality guidelines and site-specific population monitoring to evaluate the impacts of sediment-associated contaminants in estuarine systems.

Fission yeast retrotransposon Tf1 integration is targeted to 5' ends of open reading frames.

PubMed

Behrens, R; Hayles, J; Nurse, P

2000-12-01

Target site selection of transposable elements is usually not random but involves some specificity for a DNA sequence or a DNA binding host factor. We have investigated the target site selection of the long terminal repeat-containing retrotransposon Tf1 from the fission yeast Schizosaccharomyces pombe. By monitoring induced transposition events we found that Tf1 integration sites were distributed throughout the genome. Mapping these insertions revealed that Tf1 did not integrate into open reading frames, but occurred preferentially in longer intergenic regions with integration biased towards a region 100-420 bp upstream of the translation start site. Northern blot analysis showed that transcription of genes adjacent to Tf1 insertions was not significantly changed.

Fission yeast retrotransposon Tf1 integration is targeted to 5′ ends of open reading frames

PubMed Central

Behrens, Ralf; Hayles, Jacky; Nurse, Paul

2000-01-01

Target site selection of transposable elements is usually not random but involves some specificity for a DNA sequence or a DNA binding host factor. We have investigated the target site selection of the long terminal repeat-containing retrotransposon Tf1 from the fission yeast Schizosaccharomyces pombe. By monitoring induced transposition events we found that Tf1 integration sites were distributed throughout the genome. Mapping these insertions revealed that Tf1 did not integrate into open reading frames, but occurred preferentially in longer intergenic regions with integration biased towards a region 100–420 bp upstream of the translation start site. Northern blot analysis showed that transcription of genes adjacent to Tf1 insertions was not significantly changed. PMID:11095681

Assessing Hydrologic Impacts of Land Configuration Changes Using an Integrated Hydrologic Model at the Rocky Flats Environmental Technology Site, Colorado

NASA Astrophysics Data System (ADS)

Prucha, R. H.; Dayton, C. S.; Hawley, C. M.

2002-12-01

The Rocky Flats Environmental Technology Site (RFETS) in Golden, Colorado, a former Department of Energy nuclear weapons manufacturing facility, is currently undergoing closure. The natural semi-arid interaction between surface and subsurface flow at RFETS is complex and complicated by the industrial modifications to the flow system. Using a substantial site data set, a distributed parameter, fully-integrated hydrologic model was developed to assess the hydrologic impact of different hypothetical site closure configurations on the current flow system and to better understand the integrated hydrologic behavior of the system. An integrated model with this level of detail has not been previously developed in a semi-arid area, and a unique, but comprehensive, approach was required to calibrate and validate the model. Several hypothetical scenarios were developed to simulate hydrologic effects of modifying different aspects of the site. For example, some of the simulated modifications included regrading the current land surface, changing the existing surface channel network, removing subsurface trenches and gravity drain flow systems, installing a slurry wall and geotechnical cover, changing the current vegetative cover, and converting existing buildings and pavement to permeable soil areas. The integrated flow model was developed using a rigorous physically-based code so that realistic design parameters can simulate these changes. This code also permitted evaluation of changes to complex integrated hydrologic system responses that included channelized and overland flow, pond levels, unsaturated zone storage, groundwater heads and flow directions, and integrated water balances for key areas. Results generally show that channel flow offsite decreases substantially for different scenarios, while groundwater heads generally increase within the reconfigured industrial area most of which is then discharged as evapotranspiration. These changes have significant implications to site closure and operation.

HRP2 determines the efficiency and specificity of HIV-1 integration in LEDGF/p75 knockout cells but does not contribute to the antiviral activity of a potent LEDGF/p75-binding site integrase inhibitor.

PubMed

Wang, Hao; Jurado, Kellie A; Wu, Xiaolin; Shun, Ming-Chieh; Li, Xiang; Ferris, Andrea L; Smith, Steven J; Patel, Pratiq A; Fuchs, James R; Cherepanov, Peter; Kvaratskhelia, Mamuka; Hughes, Stephen H; Engelman, Alan

2012-12-01

The binding of integrase (IN) to lens epithelium-derived growth factor (LEDGF)/p75 in large part determines the efficiency and specificity of HIV-1 integration. However, a significant residual preference for integration into active genes persists in Psip1 (the gene that encodes for LEDGF/p75) knockout (KO) cells. One other cellular protein, HRP2, harbors both the PWWP and IN-binding domains that are important for LEDGF/p75 co-factor function. To assess the role of HRP2 in HIV-1 integration, cells generated from Hdgfrp2 (the gene that encodes for HRP2) and Psip1/Hdgfrp2 KO mice were infected alongside matched control cells. HRP2 depleted cells supported normal infection, while disruption of Hdgfrp2 in Psip1 KO cells yielded additional defects in the efficiency and specificity of integration. These deficits were largely restored by ectopic expression of either LEDGF/p75 or HRP2. The double-KO cells nevertheless supported residual integration into genes, indicating that IN and/or other host factors contribute to integration specificity in the absence of LEDGF/p75 and HRP2. Psip1 KO significantly increased the potency of an allosteric inhibitor that binds the LEDGF/p75 binding site on IN, a result that was not significantly altered by Hdgfrp2 disruption. These findings help to rule out the host factor-IN interactions as the primary antiviral targets of LEDGF/p75-binding site IN inhibitors.

Musite, a tool for global prediction of general and kinase-specific phosphorylation sites.

PubMed

Gao, Jianjiong; Thelen, Jay J; Dunker, A Keith; Xu, Dong

2010-12-01

Reversible protein phosphorylation is one of the most pervasive post-translational modifications, regulating diverse cellular processes in various organisms. High throughput experimental studies using mass spectrometry have identified many phosphorylation sites, primarily from eukaryotes. However, the vast majority of phosphorylation sites remain undiscovered, even in well studied systems. Because mass spectrometry-based experimental approaches for identifying phosphorylation events are costly, time-consuming, and biased toward abundant proteins and proteotypic peptides, in silico prediction of phosphorylation sites is potentially a useful alternative strategy for whole proteome annotation. Because of various limitations, current phosphorylation site prediction tools were not well designed for comprehensive assessment of proteomes. Here, we present a novel software tool, Musite, specifically designed for large scale predictions of both general and kinase-specific phosphorylation sites. We collected phosphoproteomics data in multiple organisms from several reliable sources and used them to train prediction models by a comprehensive machine-learning approach that integrates local sequence similarities to known phosphorylation sites, protein disorder scores, and amino acid frequencies. Application of Musite on several proteomes yielded tens of thousands of phosphorylation site predictions at a high stringency level. Cross-validation tests show that Musite achieves some improvement over existing tools in predicting general phosphorylation sites, and it is at least comparable with those for predicting kinase-specific phosphorylation sites. In Musite V1.0, we have trained general prediction models for six organisms and kinase-specific prediction models for 13 kinases or kinase families. Although the current pretrained models were not correlated with any particular cellular conditions, Musite provides a unique functionality for training customized prediction models (including condition-specific models) from users' own data. In addition, with its easily extensible open source application programming interface, Musite is aimed at being an open platform for community-based development of machine learning-based phosphorylation site prediction applications. Musite is available at http://musite.sourceforge.net/.

Develop and test fuel cell powered on-site integrated total energy systems. Phase 3: Full-scale power plant development

NASA Technical Reports Server (NTRS)

1981-01-01

A schematic and physical layout is given for the 5kW integrated system and the development status of individual components is described. The results of using a one dimensional mathematical model of the 5kW reformer are presented. Plans for a single-tube reformer test unit for the acquisition of temperature profile data are described. Tentative specifications for a 50kW dc-to-ac inverter are listed. Performance data are given on two 3-cell stacks incorporating semiautomatic acid replenishment systems and improved electrocatalysts. A qualification test on methanol/steam reforming catalyst T2107RS is reported, including a portion in which the catalyst was deliberately poisoned with 800 ppm ethanol in the feed.

A comprehensive analysis of Helicobacter pylori plasticity zones reveals that they are integrating conjugative elements with intermediate integration specificity.

PubMed

Fischer, Wolfgang; Breithaupt, Ute; Kern, Beate; Smith, Stella I; Spicher, Carolin; Haas, Rainer

2014-04-27

The human gastric pathogen Helicobacter pylori is a paradigm for chronic bacterial infections. Its persistence in the stomach mucosa is facilitated by several mechanisms of immune evasion and immune modulation, but also by an unusual genetic variability which might account for the capability to adapt to changing environmental conditions during long-term colonization. This variability is reflected by the fact that almost each infected individual is colonized by a genetically unique strain. Strain-specific genes are dispersed throughout the genome, but clusters of genes organized as genomic islands may also collectively be present or absent. We have comparatively analysed such clusters, which are commonly termed plasticity zones, in a high number of H. pylori strains of varying geographical origin. We show that these regions contain fixed gene sets, rather than being true regions of genome plasticity, but two different types and several subtypes with partly diverging gene content can be distinguished. Their genetic diversity is incongruent with variations in the rest of the genome, suggesting that they are subject to horizontal gene transfer within H. pylori populations. We identified 40 distinct integration sites in 45 genome sequences, with a conserved heptanucleotide motif that seems to be the minimal requirement for integration. The significant number of possible integration sites, together with the requirement for a short conserved integration motif and the high level of gene conservation, indicates that these elements are best described as integrating conjugative elements (ICEs) with an intermediate integration site specificity.

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

Daley, P F

The overall objective of this project is the continued development, installation, and testing of continuous water sampling and analysis technologies for application to on-site monitoring of groundwater treatment systems and remediation sites. In a previous project, an on-line analytical system (OLAS) for multistream water sampling was installed at the Fort Ord Operable Unit 2 Groundwater Treatment System, with the objective of developing a simplified analytical method for detection of Compounds of Concern at that plant, and continuous sampling of up to twelve locations in the treatment system, from raw influent waters to treated effluent. Earlier implementations of the water samplingmore » and processing system (Analytical Sampling and Analysis Platform, A A+RT, Milpitas, CA) depended on off-line integrators that produced paper plots of chromatograms, and sent summary tables to a host computer for archiving. We developed a basic LabVIEW (National Instruments, Inc., Austin, TX) based gas chromatography control and data acquisition system that was the foundation for further development and integration with the ASAP system. Advantages of this integration include electronic archiving of all raw chromatographic data, and a flexible programming environment to support development of improved ASAP operation and automated reporting. The initial goals of integrating the preexisting LabVIEW chromatography control system with the ASAP, and demonstration of a simplified, site-specific analytical method were successfully achieved. However, although the principal objective of this system was assembly of an analytical system that would allow plant operators an up-to-the-minute view of the plant's performance, several obstacles remained. Data reduction with the base LabVIEW system was limited to peak detection and simple tabular output, patterned after commercial chromatography integrators, with compound retention times and peak areas. Preparation of calibration curves, method detection limit estimates and trend plotting were performed with spreadsheets and statistics software. Moreover, the analytical method developed was very limited in compound coverage, and unable to closely mirror the standard analytical methods promulgated by the EPA. To address these deficiencies, during this award the original equipment was operated at the OU 2-GTS to further evaluate the use of columns, commercial standard blends and other components to broaden the compound coverage of the chromatography system. A second-generation ASAP was designed and built to replace the original system at the OU 2-GTS, and include provision for introduction of internal standard compounds and surrogates into each sample analyzed. An enhanced, LabVIEW based chromatogram analysis application was written, that manages and archives chemical standards information, and provides a basis for NIST traceability for all analyses. Within this same package, all compound calibration response curves are managed, and different report formats were incorporated, that simplify trend analysis. Test results focus on operation of the original system at the OU 1 Integrated Chemical and Flow Monitoring System, at the OU 1 Fire Drill Area remediation site.« less

Systematic data ingratiation of clinical trial recruitment locations for geographic-based query and visualization

PubMed Central

Luo, Jake; Chen, Weiheng; Wu, Min; Weng, Chunhua

2018-01-01

Background Prior studies of clinical trial planning indicate that it is crucial to search and screen recruitment sites before starting to enroll participants. However, currently there is no systematic method developed to support clinical investigators to search candidate recruitment sites according to their interested clinical trial factors. Objective In this study, we aim at developing a new approach to integrating the location data of over one million heterogeneous recruitment sites that are stored in clinical trial documents. The integrated recruitment location data can be searched and visualized using a map-based information retrieval method. The method enables systematic search and analysis of recruitment sites across a large amount of clinical trials. Methods The location data of more than 1.4 million recruitment sites of over 183,000 clinical trials was normalized and integrated using a geocoding method. The integrated data can be used to support geographic information retrieval of recruitment sites. Additionally, the information of over 6000 clinical trial target disease conditions and close to 4000 interventions was also integrated into the system and linked to the recruitment locations. Such data integration enabled the construction of a novel map-based query system. The system will allow clinical investigators to search and visualize candidate recruitment sites for clinical trials based on target conditions and interventions. Results The evaluation results showed that the coverage of the geographic location mapping for the 1.4 million recruitment sites was 99.8%. The evaluation of 200 randomly retrieved recruitment sites showed that the correctness of geographic information mapping was 96.5%. The recruitment intensities of the top 30 countries were also retrieved and analyzed. The data analysis results indicated that the recruitment intensity varied significantly across different countries and geographic areas. Conclusion This study contributed a new data processing framework to extract and integrate the location data of heterogeneous recruitment sites from clinical trial documents. The developed system can support effective retrieval and analysis of potential recruitment sites using target clinical trial factors. PMID:29132636

Systematic data ingratiation of clinical trial recruitment locations for geographic-based query and visualization.

PubMed

Luo, Jake; Chen, Weiheng; Wu, Min; Weng, Chunhua

2017-12-01

Prior studies of clinical trial planning indicate that it is crucial to search and screen recruitment sites before starting to enroll participants. However, currently there is no systematic method developed to support clinical investigators to search candidate recruitment sites according to their interested clinical trial factors. In this study, we aim at developing a new approach to integrating the location data of over one million heterogeneous recruitment sites that are stored in clinical trial documents. The integrated recruitment location data can be searched and visualized using a map-based information retrieval method. The method enables systematic search and analysis of recruitment sites across a large amount of clinical trials. The location data of more than 1.4 million recruitment sites of over 183,000 clinical trials was normalized and integrated using a geocoding method. The integrated data can be used to support geographic information retrieval of recruitment sites. Additionally, the information of over 6000 clinical trial target disease conditions and close to 4000 interventions was also integrated into the system and linked to the recruitment locations. Such data integration enabled the construction of a novel map-based query system. The system will allow clinical investigators to search and visualize candidate recruitment sites for clinical trials based on target conditions and interventions. The evaluation results showed that the coverage of the geographic location mapping for the 1.4 million recruitment sites was 99.8%. The evaluation of 200 randomly retrieved recruitment sites showed that the correctness of geographic information mapping was 96.5%. The recruitment intensities of the top 30 countries were also retrieved and analyzed. The data analysis results indicated that the recruitment intensity varied significantly across different countries and geographic areas. This study contributed a new data processing framework to extract and integrate the location data of heterogeneous recruitment sites from clinical trial documents. The developed system can support effective retrieval and analysis of potential recruitment sites using target clinical trial factors. Copyright © 2017 Elsevier B.V. All rights reserved.

78 FR 12259 - Unmanned Aircraft System Test Site Program

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-22

...-0061] Unmanned Aircraft System Test Site Program AGENCY: Federal Aviation Administration (FAA), DOT... Defense, develop a test site program for the integration of unmanned aircraft systems in to the National Airspace System. The overall purpose of this test site program is to develop a body of data and operational...

Adenosine and inflammation: what's new on the horizon?

PubMed

Antonioli, Luca; Csóka, Balázs; Fornai, Matteo; Colucci, Rocchina; Kókai, Endre; Blandizzi, Corrado; Haskó, György

2014-08-01

Adenosine contributes to the maintenance of tissue integrity by modulating the immune system. Encouraging results have emerged with adenosine receptor ligands for the management of several inflammatory conditions in preclinical and clinical settings. However, therapeutic applications of these drugs are sometimes complicated by the occurrence of serious adverse effects. The scientific community is making intensive efforts to design novel adenosine receptor ligands endowed with greater selectivity or to develop innovative compounds acting as allosteric receptor modulators. In parallel, research is focusing on novel pharmacological entities (designated as adenosine-regulating agents) that can increase, in a site- and event-specific manner, adenosine concentrations at the inflammatory site, thereby minimizing the adverse systemic effects of adenosine. Copyright © 2014 Elsevier Ltd. All rights reserved.

Progress of targeted genome modification approaches in higher plants.

PubMed

Cardi, Teodoro; Neal Stewart, C

2016-07-01

Transgene integration in plants is based on illegitimate recombination between non-homologous sequences. The low control of integration site and number of (trans/cis)gene copies might have negative consequences on the expression of transferred genes and their insertion within endogenous coding sequences. The first experiments conducted to use precise homologous recombination for gene integration commenced soon after the first demonstration that transgenic plants could be produced. Modern transgene targeting categories used in plant biology are: (a) homologous recombination-dependent gene targeting; (b) recombinase-mediated site-specific gene integration; (c) oligonucleotide-directed mutagenesis; (d) nuclease-mediated site-specific genome modifications. New tools enable precise gene replacement or stacking with exogenous sequences and targeted mutagenesis of endogeneous sequences. The possibility to engineer chimeric designer nucleases, which are able to target virtually any genomic site, and use them for inducing double-strand breaks in host DNA create new opportunities for both applied plant breeding and functional genomics. CRISPR is the most recent technology available for precise genome editing. Its rapid adoption in biological research is based on its inherent simplicity and efficacy. Its utilization, however, depends on available sequence information, especially for genome-wide analysis. We will review the approaches used for genome modification, specifically those for affecting gene integration and modification in higher plants. For each approach, the advantages and limitations will be noted. We also will speculate on how their actual commercial development and implementation in plant breeding will be affected by governmental regulations.

Strategic deployment of CHO expression platforms to deliver Pfizer's Monoclonal Antibody Portfolio.

PubMed

Scarcelli, John J; Shang, Tanya Q; Iskra, Tim; Allen, Martin J; Zhang, Lin

2017-11-01

Development of stable cell lines for expression of large-molecule therapeutics represents a significant portion of the time and effort required to advance a molecule to enabling regulatory toxicology studies and clinical evaluation. Our development strategy employs two different approaches for cell line development based on the needs of a particular project: a random integration approach for projects where high-level expression is critical, and a site-specific integration approach for projects in which speed and reduced employee time spend is a necessity. Here we describe both our random integration and site-specific integration platforms and their applications in support of monoclonal antibody development and production. We also compare product quality attributes of monoclonal antibodies produced with a nonclonal cell pool or clonal cell lines derived from the two platforms. Our data suggests that material source (pools vs. clones) does not significantly alter the examined product quality attributes. Our current practice is to leverage this observation with our site-specific integration platform, where material generated from cell pools is used for an early molecular assessment of a given candidate to make informed decisions around development strategy. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1463-1467, 2017. © 2017 American Institute of Chemical Engineers.

The salinity gradient power generating system integrated into the seawater desalination system

NASA Astrophysics Data System (ADS)

Zhu, Yongqiang; Wang, Wanjun; Cai, Bingqian; Hao, Jiacheng; Xia, Ruihua

2017-01-01

Seawater desalination is an important way to solve the problem of fresh water shortage. Low energy efficiency and high cost are disadvantages existing in seawater desalination. With huge reserve and the highest energy density among different types of marine energy, salinity gradient energy has a bright application prospect. The promotion of traditional salinity gradient power generating systems is hindered by its low efficiency and specific requirements on site selection. This paper proposes a salinity gradient power generating system integrated into the seawater desalination system which combines the salinity gradient power generating system and the seawater desalination system aiming to remedy the aforementioned deficiency and could serve as references for future seawater desalination and salinity gradient energy exploitation. The paper elaborates on the operating principles of the system, analyzes the detailed working process, and estimates the energy output and consumption of the system. It is proved that with appropriate design, the energy output of the salinity gradient power generating system can satisfy the demand of the seawater desalination system.

Production of transgenic cattle highly expressing human serum albumin in milk by phiC31 integrase-mediated gene delivery.

PubMed

Luo, Yan; Wang, Yongsheng; Liu, Jun; Lan, Hui; Shao, Minghao; Yu, Yuan; Quan, Fusheng; Zhang, Yong

2015-10-01

Transgenic cattle expressing high levels of recombinant human serum albumin (HSA) in their milk may as an alternative source for commercial production. Our objective was to produce transgenic cattle highly expressing HSA in milk by using phiC31 integrase system and somatic cell nuclear transfer (SCNT). The mammary-specific expression plasmid pIACH(-), containing the attB recognition site for phiC31 integrase, were co-transfected with integrase expression plasmid pCMVInt into bovine fetal fibroblast cells (BFFs). PhiC31 integrase-mediated integrations in genome of BFFs were screened by nested inverse PCR. After analysis of sequence of the PCR products, 46.0% (23/50) of the both attB-genome junction sites (attL and attR) were confirmed, and four pseudo attP sites were identified. The integration rates in BF3, BF11, BF19 and BF4 sites were 4.0% (2/50), 6.0% (3/50), 16.0% (8/50) and 20.0% (10/50), respectively. BF3 is located in the bovine chromosome 3 collagen alpha-3 (VI) chain isomer 2 gene, while the other three sites are located in the non-coding region. The transgenic cell lines from BF11, BF19 and BF4 sites were used as donors for SCNT. Two calves from transgenic cells BF19 were born, one died within a few hours after birth, and another calf survived healthy. PCR and Southern blot analysis revealed integration of the transgene in the genome of cloned calves. The nested reverse PCR confirmed that the integration site in cloned calves was identical to the donor cells. The western blotting assessment indicated that recombinant HSA was expressed in the milk of transgenic cattle and the expression level was about 4-8 mg/mL. The present study demonstrated that phiC31 integrase system was an efficient and safety gene delivery tool for producing HSA transgenic cattle. The production of recombinant HSA in the milk of cattle may provide a large-scale and cost-effective resource.

Distinct oxidative cleavage and modification of bovine [Cu-Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule

PubMed Central

Uehara, Hiroshi; Luo, Shen; Aryal, Baikuntha; Levine, Rodney L.; Rao, V. Ashutosh

2016-01-01

We investigated the combined effect of ascorbate and copper [Asc/Cu(II)] on the integrity of bovine [Cu-Zn]-superoxide dismutase (bSOD1) as a model system to study the metal catalyzed oxidation (MCO) and fragmentation of proteins. We found Asc/Cu(II) mediates specific cleavage of bSOD1 and generates 12.5 and 3.2 kDa fragments in addition to oxidation/carbonylation of the protein. The effect of other tested transition metals, a metal chelator, and hydrogen peroxide on the cleavage and oxidation indicated that binding of copper to a previously unknown site on SOD1 is responsible for the Asc/Cu(II) specific cleavage and oxidation. We utilized tandem mass spectrometry to identify the specific cleavage sites of Asc/Cu(II)-treated bSOD1. Analyses of tryptic- and AspN-peptides have demonstrated the cleavage to occur at Gly31 with peptide bond breakage with Thr30 and Ser32 through diamide and α-amidation pathways, respectively. The three-dimensional structure of bSOD1 reveals the imidazole ring of His19 localized within 5 Angstrom from the α-carbon of Gly31 providing a structural basis that copper ion, most likely coordinated by His19, catalyzes the specific cleavage reaction. PMID:26872685

Distinct oxidative cleavage and modification of bovine [Cu- Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule.

PubMed

Uehara, Hiroshi; Luo, Shen; Aryal, Baikuntha; Levine, Rodney L; Rao, V Ashutosh

2016-05-01

We investigated the combined effect of ascorbate and copper [Asc/Cu(II)] on the integrity of bovine [Cu-Zn]-superoxide dismutase (bSOD1) as a model system to study the metal catalyzed oxidation (MCO) and fragmentation of proteins. We found Asc/Cu(II) mediates specific cleavage of bSOD1 and generates 12.5 and 3.2kDa fragments in addition to oxidation/carbonylation of the protein. The effect of other tested transition metals, a metal chelator, and hydrogen peroxide on the cleavage and oxidation indicated that binding of copper to a previously unknown site on SOD1 is responsible for the Asc/Cu(II) specific cleavage and oxidation. We utilized tandem mass spectrometry to identify the specific cleavage sites of Asc/Cu(II)-treated bSOD1. Analyses of tryptic- and AspN-peptides have demonstrated the cleavage to occur at Gly31 with peptide bond breakage with Thr30 and Ser32 through diamide and α-amidation pathways, respectively. The three-dimensional structure of bSOD1 reveals the imidazole ring of His19 localized within 5Å from the α-carbon of Gly31 providing a structural basis that copper ion, most likely coordinated by His19, catalyzes the specific cleavage reaction. Published by Elsevier Inc.

Enhancers Are Major Targets for Murine Leukemia Virus Vector Integration

PubMed Central

De Ravin, Suk See; Su, Ling; Theobald, Narda; Choi, Uimook; Macpherson, Janet L.; Poidinger, Michael; Symonds, Geoff; Pond, Susan M.; Ferris, Andrea L.; Hughes, Stephen H.

2014-01-01

ABSTRACT Retroviral vectors have been used in successful gene therapies. However, in some patients, insertional mutagenesis led to leukemia or myelodysplasia. Both the strong promoter/enhancer elements in the long terminal repeats (LTRs) of murine leukemia virus (MLV)-based vectors and the vector-specific integration site preferences played an important role in these adverse clinical events. MLV integration is known to prefer regions in or near transcription start sites (TSS). Recently, BET family proteins were shown to be the major cellular proteins responsible for targeting MLV integration. Although MLV integration sites are significantly enriched at TSS, only a small fraction of the MLV integration sites (<15%) occur in this region. To resolve this apparent discrepancy, we created a high-resolution genome-wide integration map of more than one million integration sites from CD34+ hematopoietic stem cells transduced with a clinically relevant MLV-based vector. The integration sites form ∼60,000 tight clusters. These clusters comprise ∼1.9% of the genome. The vast majority (87%) of the integration sites are located within histone H3K4me1 islands, a hallmark of enhancers. The majority of these clusters also have H3K27ac histone modifications, which mark active enhancers. The enhancers of some oncogenes, including LMO2, are highly preferred targets for integration without in vivo selection. IMPORTANCE We show that active enhancer regions are the major targets for MLV integration; this means that MLV preferentially integrates in regions that are favorable for viral gene expression in a variety of cell types. The results provide insights for MLV integration target site selection and also explain the high risk of insertional mutagenesis that is associated with gene therapy trials using MLV vectors. PMID:24501411

Study of component technologies for fuel cell on-site integrated energy system. Volume 2: Appendices

NASA Technical Reports Server (NTRS)

Lee, W. D.; Mathias, S.

1980-01-01

This data base catalogue was compiled in order to facilitate the analysis of various on site integrated energy system with fuel cell power plants. The catalogue is divided into two sections. The first characterizes individual components in terms of their performance profiles as a function of design parameters. The second characterizes total heating and cooling systems in terms of energy output as a function of input and control variables. The integrated fuel cell systems diagrams and the computer analysis of systems are included as well as the cash flows series for baseline systems.

Recording, monitoring and managing the conservation of historic sites: a new application for BGS·SIGMA

NASA Astrophysics Data System (ADS)

Tracey, Emily; Smith, Nichola; Lawrie, Ken

2017-04-01

The principles behind, and the methods of, digital data capture can be applied across many scientific, and other, disciplines, as can be demonstrated by the use of a custom modified version of the British Geological Survey's System for Integrated Geoscience Mapping, (BGS·SIGMA), for the capture of data for use in the conservation of Scottish built heritage. Historic Environment Scotland (HES), an executive agency of the Scottish Government charged with safeguarding the nation's historic environment, is directly responsible for 345 sites of national significance, most of which are built from stone. In common with many other heritage organisations, HES needs a system that can capture, store and present conservation, maintenance and condition indicator information for single or multiple historic sites; this system would then be used to better target and plan effective programmes of maintenance and repair. To meet this need, the British Geological Survey (BGS) has worked with HES to develop an integrated digital site assessment system that provides a refined survey process for stone-built (and other) historic sites. Based on BGS·SIGMA—an integrated workflow underpinned by a geo-spatial platform for data capture and interpretation—the new system is built on top of ESRI's ArcGIS software, and underpinned by a relational database. Users can, in the field or in the office, populate custom-built data entry forms to record maintenance issues and repair specifications for architectural elements ranging from individual blocks of stone to entire building elevations. Photographs, sketches, and digital documents can be linked to architectural elements to enhance the usability of the data. Predetermined data fields and supporting dictionaries constrain the input parameters, ensuring a high degree of standardisation in the datasets and, therefore, enabling highly consistent data extraction and querying. The GIS presentation of the data provides a powerful and versatile planning tool for scheduling works, specifying materials, identifying the skills needed for repairs, and allocating resources more effectively and efficiently. Physical alterations and changes in the overall condition of a single site, or a group of sites can be monitored accurately over time by repeating the original survey (e.g. every 5 years). Other datasets can be linked to the database and other geospatially referenced datasets can be superimposed in GIS, adding considerably to the scope and utility of the system. The system can be applied to any geospatially referenced object in a wide range of situations thus providing many potential applications in conservation, archaeology and other related fields.

Event-specific real-time detection and quantification of genetically modified Roundup Ready soybean.

PubMed

Huang, Chia-Chia; Pan, Tzu-Ming

2005-05-18

The event-specific real-time detection and quantification of Roundup Ready soybean (RRS) using an ABI PRISM 7700 sequence detection system with light upon extension (LUX) primer was developed in this study. The event-specific primers were designed, targeting the junction of the RRS 5' integration site and the endogenous gene lectin1. Then, a standard reference plasmid was constructed that carried both of the targeted sequences for quantitative analysis. The detection limit of the LUX real-time PCR system was 0.05 ng of 100% RRS genomic DNA, which was equal to 20.5 copies. The range of quantification was from 0.1 to 100%. The sensitivity and range of quantification successfully met the requirement of the labeling rules in the European Union and Taiwan.

piggyBac Transposon-Mediated Transgenesis in the Apicomplexan Parasite Eimeria tenella

PubMed Central

Su, Huali; Liu, Xianyong; Yan, Wenchao; Shi, Tuanyuan; Zhao, Xinxin; Blake, Damer P.; Tomley, Fiona M.; Suo, Xun

2012-01-01

piggyBac, a type II transposon that is useful for efficient transgenesis and insertional mutagenesis, has been used for effective and stable transfection in a wide variety of organisms. In this study we investigate the potential use of the piggyBac transposon system for forward genetics studies in the apicomplexan parasite Eimeria tenella. Using the restriction enzyme-mediated integration (REMI) method, E. tenella sporozoites were electroporated with a donor plasmid containing the enhanced yellow fluorescent protein (EYFP) gene flanked by piggyBac inverted terminal repeats (ITRs), an Asc I-linearized helper plasmid containing the transposase gene and the restriction enzyme Asc I. Subsequently, electroporated sporozoites were inoculated into chickens via the cloacal route and transfected progeny oocysts expressing EYFP were sorted by flow cytometry. A transgenic E. tenella population was selected by successive in vivo passage. Southern-blotting analysis showed that exogenous DNA containing the EYFP gene was integrated into the parasite genome at a limited number of integration sites and that the inserted part of the donor plasmid was the fragment located between the 5′ and 3′ ITRs as indicated by primer-specific PCR screening. Genome walking revealed that the insertion sites were TTAA-specific, which is consistent with the transposition characteristics of piggyBac. PMID:22768223

Diagnostic accuracy of integrated intravascular ultrasound and optical coherence tomography (IVUS-OCT) system for coronary plaque characterization

NASA Astrophysics Data System (ADS)

Li, Jiawen; Ma, Teng; Mohar, Dilbahar; Correa, Adrian; Minami, Hataka; Jing, Joseph; Zhou, Qifa; Patel, Pranav M.; Chen, Zhongping

2014-03-01

Intravascular ultrasound (IVUS) imaging and optical coherence tomography (OCT), two commonly used intracoronary imaging modalities, play important roles in plaque evaluation. The combined use of IVUS (to visualize the entire plaque volume) and OCT (to quantify the thickness of the plaque cap, if any) is hypothesized to increase plaque diagnostic accuracy. Our group has developed a fully-integrated dual-modality IVUS-OCT imaging system and 3.6F catheter for simultaneous IVUS-OCT imaging with a high resolution and deep penetration depth. However, the diagnostic accuracy of an integrated IVUS-OCT system has not been investigated. In this study, we imaged 175 coronary artery sites (241 regions of interest) from 20 cadavers using our previous reported integrated IVUS-OCT system. IVUS-OCT images were read by two skilled interventional cardiologists. Each region of interest was classified as either calcification, lipid pool or fibrosis. Comparing the diagnosis by cardiologists using IVUSOCT images with the diagnosis by the pathologist, we calculated the sensitivity and specificity for characterization of calcification, lipid pool or fibrosis with this integrated system. In vitro imaging of cadaver coronary specimens demonstrated the complementary nature of these two modalities for plaques classification. A higher accuracy was shown than using a single modality alone.

Gene-specific cell labeling using MiMIC transposons

PubMed Central

Gnerer, Joshua P.; Venken, Koen J. T.; Dierick, Herman A.

2015-01-01

Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family. PMID:25712101

An efficient strategy for producing a stable, replaceable, highly efficient transgene expression system in silkworm, Bombyx mori

PubMed Central

Long, Dingpei; Lu, Weijian; Zhang, Yuli; Bi, Lihui; Xiang, Zhonghuai; Zhao, Aichun

2015-01-01

We developed an efficient strategy that combines a method for the post-integration elimination of all transposon sequences, a site-specific recombination system, and an optimized fibroin H-chain expression system to produce a stable, replaceable, highly efficient transgene expression system in the silkworm (Bombyx mori) that overcomes the disadvantages of random insertion and post-integration instability of transposons. Here, we generated four different transgenic silkworm strains, and of one the transgenic strains, designated TS1-RgG2, with up to 16% (w/w) of the target protein in the cocoons, was selected. The subsequent elimination of all the transposon sequences from TS1-RgG2 was completed by the heat-shock-induced expression of the transposase in vivo. The resulting transgenic silkworm strain was designated TS3-g2 and contained only the attP-flanked optimized fibroin H-chain expression cassette in its genome. A phiC31/att-system-based recombinase-mediated cassette exchange (RMCE) method could be used to integrate other genes of interest into the same genome locus between the attP sites in TS3-g2. Controlling for position effects with phiC31-mediated RMCE will also allow the optimization of exogenous protein expression and fine gene function analyses in the silkworm. The strategy developed here is also applicable to other lepidopteran insects, to improve the ecological safety of transgenic strains in biocontrol programs. PMID:25739894

Assessment of fish assemblages and minimum sampling effort required to determine botic integrity of large rivers in southern Idaho, 2002

USGS Publications Warehouse

Maret, Terry R.; Ott, D.S.

2004-01-01

width was determined to be sufficient for collecting an adequate number of fish to estimate species richness and evaluate biotic integrity. At most sites, about 250 fish were needed to effectively represent 95 percent of the species present. Fifty-three percent of the sites assessed, using an IBI developed specifically for large Idaho rivers, received scores of less than 50, indicating poor biotic integrity.

Managing Data, Provenance and Chaos through Standardization and Automation at the Georgia Coastal Ecosystems LTER Site

NASA Astrophysics Data System (ADS)

Sheldon, W.

2013-12-01

Managing data for a large, multidisciplinary research program such as a Long Term Ecological Research (LTER) site is a significant challenge, but also presents unique opportunities for data stewardship. LTER research is conducted within multiple organizational frameworks (i.e. a specific LTER site as well as the broader LTER network), and addresses both specific goals defined in an NSF proposal as well as broader goals of the network; therefore, every LTER data can be linked to rich contextual information to guide interpretation and comparison. The challenge is how to link the data to this wealth of contextual metadata. At the Georgia Coastal Ecosystems LTER we developed an integrated information management system (GCE-IMS) to manage, archive and distribute data, metadata and other research products as well as manage project logistics, administration and governance (figure 1). This system allows us to store all project information in one place, and provide dynamic links through web applications and services to ensure content is always up to date on the web as well as in data set metadata. The database model supports tracking changes over time in personnel roles, projects and governance decisions, allowing these databases to serve as canonical sources of project history. Storing project information in a central database has also allowed us to standardize both the formatting and content of critical project information, including personnel names, roles, keywords, place names, attribute names, units, and instrumentation, providing consistency and improving data and metadata comparability. Lookup services for these standard terms also simplify data entry in web and database interfaces. We have also coupled the GCE-IMS to our MATLAB- and Python-based data processing tools (i.e. through database connections) to automate metadata generation and packaging of tabular and GIS data products for distribution. Data processing history is automatically tracked throughout the data lifecycle, from initial import through quality control, revision and integration by our data processing system (GCE Data Toolbox for MATLAB), and included in metadata for versioned data products. This high level of automation and system integration has proven very effective in managing the chaos and scalability of our information management program.

Desert Research and Technology Studies (DRATS) Traverse Planning

NASA Technical Reports Server (NTRS)

Horz, Friedrich

2012-01-01

Slide 1] The Desert Research and Technology Studies (DRATS) include large scale field tests of manned lunar surface exploration systems; these tests are sponsored by the Director s Office of Integration (DOI) [sic, Directorate Integration Office (DIO)] within the Constellation Program and they include geological exploration objectives along well designed traverses. These traverses are designed by the Traverse Team, an ad hoc group of some 10 geologists form NASA and academia, as well as experts in mission operation who define the operational constraints applicable to specific simulation scenarios. [Slide 2] These DRATS/DOI tests focus on 1) the performance of major surface systems, such as rovers, mobile habitats, communication architecture, navigation tools, earth-moving equipment, unmanned reconnaissance robots etc. under realistic field conditions and 2) the development of operational concepts that integrate all of these systems into a single, optimized operation. The participation of science is currently concentrating on geological sciences, with the objective of developing suitable tools and documentation protocols to sample representative rocks for Earth return, and to generate some conceptual understanding of the ground support structure that will be needed for the real time science-support of a lunar surface crew. [Slide 3] Major surface systems exercised in the June 2008 analog tests at the Moses Lake site, WA. [Upper left] The Chariot Rover (developed at Johnson Space Center) is an unpressurized vehicle driven by fully suited crews. [Upper right] Mobile Habitat provided by the Jet Propulsion Laboratory. Chariot is the more nimble and mobile vehicle and the idea is to drive the habitat remotely to some rendezvous place where Chariot would catch up - after a lengthy traverse - at the end of the day. [Lower left] The K-10 remotely operated robot (provided by NASA Ames Research Center) conducting scientific/geologic reconnaissance of the prospective traverse region, locating specific sites for more detailed exploration by Chariot and its crew. [Lower right] This earth-moving equipment (provided by NASA KSC) can be attached to Chariot and is envisioned to, for example, level an outpost site or to mine lunar soi

Integrating soil information into canopy sensor algorithms for improved corn nitrogen rate recommendation

USDA-ARS?s Scientific Manuscript database

Crop canopy sensors have proven effective at determining site-specific nitrogen (N) needs, but several Midwest states use different algorithms to predict site-specific N need. The objective of this research was to determine if soil information can be used to improve the Missouri canopy sensor algori...

D Integrated Methodologies for the Documentation and the Virtual Reconstruction of AN Archaeological Site

NASA Astrophysics Data System (ADS)

Balletti, C.; Guerra, F.; Scocca, V.; Gottardi, C.

2015-02-01

Highly accurate documentation and 3D reconstructions are fundamental for analyses and further interpretations in archaeology. In the last years the integrated digital survey (ground-based survey methods and UAV photogrammetry) has confirmed its main role in the documentation and comprehension of excavation contexts, thanks to instrumental and methodological development concerning the on site data acquisition. The specific aim of the project, reported in this paper and realized by the Laboratory of Photogrammetry of the IUAV University of Venice, is to check different acquisition systems and their effectiveness test, considering each methodology individually or integrated. This research focuses on the awareness that the integration of different survey's methodologies can as a matter of fact increase the representative efficacy of the final representations; these are based on a wider and verified set of georeferenced metric data. Particularly the methods' integration allows reducing or neutralizing issues related to composite and complex objects' survey, since the most appropriate tools and techniques can be chosen considering the characteristics of each part of an archaeological site (i.e. urban structures, architectural monuments, small findings). This paper describes the experience in several sites of the municipality of Sepino (Molise, Italy), where the 3d digital acquisition of cities and structure of monuments, sometimes hard to reach, was realized using active and passive techniques (rage-based and image based methods). This acquisition was planned in order to obtain not only the basic support for interpretation analysis, but also to achieve models of the actual state of conservation of the site on which some reconstructive hypotheses can be based on. Laser scanning data were merged with Structure from Motion techniques' clouds into the same reference system, given by a topographical and GPS survey. These 3d models are not only the final results of the metric survey, but also the starting point for the whole reconstruction of the city and its urban context, from the research point of view. This reconstruction process will concern even some areas that have not yet been excavated, where the application of procedural modelling can offer an important support to the reconstructive hypothesis.

AGIS: Integration of new technologies used in ATLAS Distributed Computing

NASA Astrophysics Data System (ADS)

Anisenkov, Alexey; Di Girolamo, Alessandro; Alandes Pradillo, Maria

2017-10-01

The variety of the ATLAS Distributed Computing infrastructure requires a central information system to define the topology of computing resources and to store different parameters and configuration data which are needed by various ATLAS software components. The ATLAS Grid Information System (AGIS) is the system designed to integrate configuration and status information about resources, services and topology of the computing infrastructure used by ATLAS Distributed Computing applications and services. Being an intermediate middleware system between clients and external information sources (like central BDII, GOCDB, MyOSG), AGIS defines the relations between experiment specific used resources and physical distributed computing capabilities. Being in production during LHC Runl AGIS became the central information system for Distributed Computing in ATLAS and it is continuously evolving to fulfil new user requests, enable enhanced operations and follow the extension of the ATLAS Computing model. The ATLAS Computing model and data structures used by Distributed Computing applications and services are continuously evolving and trend to fit newer requirements from ADC community. In this note, we describe the evolution and the recent developments of AGIS functionalities, related to integration of new technologies recently become widely used in ATLAS Computing, like flexible computing utilization of opportunistic Cloud and HPC resources, ObjectStore services integration for Distributed Data Management (Rucio) and ATLAS workload management (PanDA) systems, unified storage protocols declaration required for PandDA Pilot site movers and others. The improvements of information model and general updates are also shown, in particular we explain how other collaborations outside ATLAS could benefit the system as a computing resources information catalogue. AGIS is evolving towards a common information system, not coupled to a specific experiment.

Comparison of Computer-based Clinical Decision Support Systems and Content for Diabetes Mellitus.

PubMed

Kantor, M; Wright, A; Burton, M; Fraser, G; Krall, M; Maviglia, S; Mohammed-Rajput, N; Simonaitis, L; Sonnenberg, F; Middleton, B

2011-01-01

Computer-based clinical decision support (CDS) systems have been shown to improve quality of care and workflow efficiency, and health care reform legislation relies on electronic health records and CDS systems to improve the cost and quality of health care in the United States; however, the heterogeneity of CDS content and infrastructure of CDS systems across sites is not well known. We aimed to determine the scope of CDS content in diabetes care at six sites, assess the capabilities of CDS in use at these sites, characterize the scope of CDS infrastructure at these sites, and determine how the sites use CDS beyond individual patient care in order to identify characteristics of CDS systems and content that have been successfully implemented in diabetes care. We compared CDS systems in six collaborating sites of the Clinical Decision Support Consortium. We gathered CDS content on care for patients with diabetes mellitus and surveyed institutions on characteristics of their site, the infrastructure of CDS at these sites, and the capabilities of CDS at these sites. The approach to CDS and the characteristics of CDS content varied among sites. Some commonalities included providing customizability by role or user, applying sophisticated exclusion criteria, and using CDS automatically at the time of decision-making. Many messages were actionable recommendations. Most sites had monitoring rules (e.g. assessing hemoglobin A1c), but few had rules to diagnose diabetes or suggest specific treatments. All sites had numerous prevention rules including reminders for providing eye examinations, influenza vaccines, lipid screenings, nephropathy screenings, and pneumococcal vaccines. Computer-based CDS systems vary widely across sites in content and scope, but both institution-created and purchased systems had many similar features and functionality, such as integration of alerts and reminders into the decision-making workflow of the provider and providing messages that are actionable recommendations.

A Geographical Information System Based Approach for Integrated Strategies of Tick Surveillance and Control in the Peri-Urban Natural Reserve of Monte Pellegrino (Palermo, Southern Italy)

PubMed Central

Torina, Alessandra; Blanda, Marcellocalogero; Auteri, Michelangelo; La Russa, Francesco; Scimeca, Salvatore; D’Agostino, Rosalia; Disclafani, Rosaria; Currò, Vittoria; Caracappa, Santo

2018-01-01

Ticks (Acari: Ixodidae) are bloodsucking arthropods involved in pathogen transmission in animals and humans. Tick activity depends on various ecological factors such as vegetation, hosts, and temperature. The aim of this study was to analyse the spatial/temporal distribution of ticks in six sites within a peri-urban area of Palermo (Natural Reserve of Monte Pellegrino) and correlate it with field data using Geographical Information System (GIS) data. A total of 3092 ticks were gathered via dragging method from June 2012 to May 2014. The species collected were: Ixodes ventalloi (46.09%), Hyalomma lusitanicum (19.99%), Rhipicephalus sanguineus (17.34%), Rhipicephalus pusillus (16.11%), Haemaphisalis sulcata (0.36%), Dermacentor marginatus (0.10%), and Rhipicephalus turanicus (0.03%). GIS analysis revealed environmental characteristics of each site, and abundance of each tick species was analysed in relation to time (monthly trend) and space (site-specific abundance). A relevant presence of I. ventalloi in site 2 and H. lusitanicum in site 5 was observed, suggesting the possible exposure of animals and humans to tick-borne pathogens. Our study shows the importance of surveillance of ticks in peri-urban areas and the useful implementation of GIS analysis in vector ecology; studies on temporal and spatial distribution of ticks correlated to GIS-based ecological analysis represent an integrated strategy for decision support in public health. PMID:29495440

STK Integrated Message Production List Editor (SIMPLE) for CEO Operations

NASA Technical Reports Server (NTRS)

Trenchard, Mike; Heydorn, James

2014-01-01

Late in fiscal year 2011, the Crew Earth Observations (CEO) team was tasked to upgrade and replace its mission planning and mission operations software systems, which were developed in the Space Shuttle era of the 1980s and 1990s. The impetuses for this change were the planned transition of all workstations to the Windows 7 64-bit operating system and the desire for more efficient and effective use of Satellite Tool Kit (STK) software required for reliable International Space Station (ISS) Earth location tracking. An additional requirement of this new system was the use of the same SQL database of CEO science sites from the SMMS, which was also being developed. STK Integrated Message Production List Editor (SIMPLE) is the essential, all-in-one tool now used by CEO staff to perform daily ISS mission planning to meet its requirement to acquire astronaut photography of specific sites on Earth. The sites are part of a managed, long-term database that has been defined and developed for scientific, educational, and public interest. SIMPLE's end product is a set of basic time and location data computed for an operator-selected set of targets that the ISS crew will be asked to photograph (photography is typically planned 12 to 36 hours out). The CEO operator uses SIMPLE to (a) specify a payload operations planning period; (b) acquire and validate the best available ephemeris data (vectors) for the ISS during the planning period; (c) ingest and display mission-specific site information from the CEO database; (d) identify and display potential current dynamic event targets as map features; (e) compute and display time and location information for each target; (f) screen and select targets based on known crew availability constraints, obliquity constraints, and real-time evaluated constraints to target visibility due to illumination (sun elevation) and atmospheric conditions (weather); and finally (g) incorporate basic, computed time and location information for each selected target into the daily CEO Target List product (message) for submission to ISS payload planning and integration teams for their review and approval prior to uplink. SIMPLE requires and uses the following resources: an ISS mission planning period Greenwich Mean Time start date/time and end date/time), the best available ISS mission ephemeris data (vectors) for that planning period, the STK software package configured for the ISS, and an ISS mission-specific subset of the CEO sites database. The primary advantages realized by the development and implementation of SIMPLE into the CEO payload operations support activity are a smooth transition to the Windows 7 operating system upon scheduled workstation refresh; streamlining of the input and verification of the current ISS ephemeris (vector data); seamless incorporation of selected contents of the SQL database of science sites; the ability to tag and display potential dynamic event opportunities on orbit track maps; simplification of the display and selection of encountered sites based on crew availability, illumination, obliquity, and weather constraints; the incorporation of high-quality mapping of the Earth with various satellite-based datasets for use in describing targets; and the ability to encapsulate and export the essential selected target elements in XML format for use by onboard Earth-location systems, such as Worldmap. SIMPLE is a carefully designed and crafted in-house software package that includes detailed help files for the user and meticulous internal documentation for future modifications. It was delivered in February 2012 for test and evaluation. Following acceptance, it was implemented for CEO mission operations support in May 2012.

Nanoparticle-Hydrogel: A Hybrid Biomaterial System for Localized Drug Delivery

PubMed Central

Gao, Weiwei; Zhang, Yue; Zhang, Qiangzhe; Zhang, Liangfang

2016-01-01

Nanoparticles have offered a unique set of properties for drug delivery including high drug loading capacity, combinatorial delivery, controlled and sustained drug release, prolonged stability and lifetime, and targeted delivery. To further enhance therapeutic index, especially for localized application, nanoparticles have been increasingly combined with hydrogels to form a hybrid biomaterial system for controlled drug delivery. Herein, we review recent progresses in engineering such nanoparticle-hydrogel hybrid system (namely ‘NP-gel’) with a particular focus on its application for localized drug delivery. Specifically, we highlight four research areas where NP-gel has shown great promises, including (1) passively controlled drug release, (2) stimuli-responsive drug delivery, (3) site-specific drug delivery, and (4) detoxification. Overall, integrating therapeutic nanoparticles with hydrogel technologies creates a unique and robust hybrid biomaterial system that enables effective localized drug delivery. PMID:26951462

Remote and terrestrial ground monitoring techniques integration for hazard assessment in mountain areas

NASA Astrophysics Data System (ADS)

Chinellato, Giulia; Kenner, Robert; Iasio, Christian; Mair, Volkmar; Mosna, David; Mulas, Marco; Phillips, Marcia; Strada, Claudia; Zischg, Andreas

2014-05-01

In high mountain regions the choice of appropriate sites for infrastructure such as roads, railways, cable cars or hydropower dams is often very limited. In parallel, the increasing demand for supply infrastructure in the Alps induces a continuous transformation of the territory. The new role played by the precautionary monitoring in the risk governance becomes fundamental and may overcome the modeling of future events, which represented so far the predominant approach to these sort of issues. Furthermore the consequence of considering methodologies alternative to those more exclusive allow to reduce costs and increasing the frequency of measurements, updating continuously the cognitive framework of existing hazard condition in most susceptible territories. The scale factor of the observed area and the multiple purpose of such regional ordinary surveys make it convenient to adopt Radar Satellite-based systems, but they need to be integrated with terrestrial systems for validation and eventual early warning purposes. Significant progress over the past decade in Remote Sensing (RS), Proximal Sensing and integration-based sensor networks systems now provide technologies, that allow to implement monitoring systems for ordinary surveys of extensive areas or regions, which are affected by active natural processes and slope instability. The Interreg project SloMove aims to provide solutions for such challenges and focuses on using remote sensing monitoring techniques for the monitoring of mass movements in two test sites, in South Tyrol (Italy) and in Grisons Canton (Switzerland). The topics faced in this project concern mass movements and slope deformation monitoring techniques, focusing mainly on the integration of multi-temporal interferometry, new generation of terrestrial technologies for differential digital terrain model elaboration provided by laser scanner (TLS), and GNSS-based topographic surveys, which are used not only for validation purpose, but also for adding value and information to the whole monitoring survey. The test sites are currently observed by an original integrated methodology specifically developed within the aim of the project. The integrated monitoring design includes reference targets for the different monitoring systems placed together on the same point or rigid foundation, to facilitate the comparison of the data and, in the operational use, to be able to switch consistently from one to the other system. The principal goal of the project is to define a shared procedure to select scalable technologies, best practices and institutional action plans more adequate to deal with different sort of hazard related to ground displacement, in densely populated mountain areas containing recreational and critical infrastructures. Keywords: integrated monitoring, multi-temporal interferometry, artificial reflectors; mass movement, SloMove.eu

Cas4-Dependent Prespacer Processing Ensures High-Fidelity Programming of CRISPR Arrays.

PubMed

Lee, Hayun; Zhou, Yi; Taylor, David W; Sashital, Dipali G

2018-04-05

CRISPR-Cas immune systems integrate short segments of foreign DNA as spacers into the host CRISPR locus to provide molecular memory of infection. Cas4 proteins are widespread in CRISPR-Cas systems and are thought to participate in spacer acquisition, although their exact function remains unknown. Here we show that Bacillus halodurans type I-C Cas4 is required for efficient prespacer processing prior to Cas1-Cas2-mediated integration. Cas4 interacts tightly with the Cas1 integrase, forming a heterohexameric complex containing two Cas1 dimers and two Cas4 subunits. In the presence of Cas1 and Cas2, Cas4 processes double-stranded substrates with long 3' overhangs through site-specific endonucleolytic cleavage. Cas4 recognizes PAM sequences within the prespacer and prevents integration of unprocessed prespacers, ensuring that only functional spacers will be integrated into the CRISPR array. Our results reveal the critical role of Cas4 in maintaining fidelity during CRISPR adaptation, providing a structural and mechanistic model for prespacer processing and integration. Copyright © 2018 Elsevier Inc. All rights reserved.

Field Testing of Environmentally Friendly Drilling System

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

David Burnett

2009-05-31

The Environmentally Friendly Drilling (EFD) program addresses new low-impact technology that reduces the footprint of drilling activities, integrates light weight drilling rigs with reduced emission engine packages, addresses on-site waste management, optimizes the systems to fit the needs of a specific development sites and provides stewardship of the environment. In addition, the program includes industry, the public, environmental organizations, and elected officials in a collaboration that addresses concerns on development of unconventional natural gas resources in environmentally sensitive areas. The EFD program provides the fundamentals to result in greater access, reasonable regulatory controls, lower development cost and reduction of themore » environmental footprint associated with operations for unconventional natural gas. Industry Sponsors have supported the program with significant financial and technical support. This final report compendium is organized into segments corresponding directly with the DOE approved scope of work for the term 2005-2009 (10 Sections). Each specific project is defined by (a) its goals, (b) its deliverable, and (c) its future direction. A web site has been established that contains all of these detailed engineering reports produced with their efforts. The goals of the project are to (1) identify critical enabling technologies for a prototype low-impact drilling system, (2) test the prototype systems in field laboratories, and (3) demonstrate the advanced technology to show how these practices would benefit the environment.« less

Genome-wide analysis of Tol2 transposon reintegration in zebrafish.

PubMed

Kondrychyn, Igor; Garcia-Lecea, Marta; Emelyanov, Alexander; Parinov, Sergey; Korzh, Vladimir

2009-09-08

Tol2, a member of the hAT family of transposons, has become a useful tool for genetic manipulation of model animals, but information about its interactions with vertebrate genomes is still limited. Furthermore, published reports on Tol2 have mainly been based on random integration of the transposon system after co-injection of a plasmid DNA harboring the transposon and a transposase mRNA. It is important to understand how Tol2 would behave upon activation after integration into the genome. We performed a large-scale enhancer trap (ET) screen and generated 338 insertions of the Tol2 transposon-based ET cassette into the zebrafish genome. These insertions were generated by remobilizing the transposon from two different donor sites in two transgenic lines. We found that 39% of Tol2 insertions occurred in transcription units, mostly into introns. Analysis of the transposon target sites revealed no strict specificity at the DNA sequence level. However, Tol2 was prone to target AT-rich regions with weak palindromic consensus sequences centered at the insertion site. Our systematic analysis of sequential remobilizations of the Tol2 transposon from two independent sites within a vertebrate genome has revealed properties such as a tendency to integrate into transcription units and into AT-rich palindrome-like sequences. This information will influence the development of various applications involving DNA transposons and Tol2 in particular.

Cleanups in My Community

EPA Pesticide Factsheets

Cleanups in My Community (CIMC) is a public web application that enables integrated access through maps, lists and search filtering to site-specific information EPA has across all cleanup programs. CIMC taps into data publicly available from EPA's EnviroFacts (RCRA Corrective Action facilities, Brownfields properties and grant areas, Superfund NPL sites, other facility data) and web services (water monitoring stations, impaired waters, emergency responses, tribal boundaries, congressional districts, etc.) and connects to other applications (e.g., Superfund's CPAD) to provide easy seamless access to site-specific cleanup information with explanatory text and within the context of related data. Data can be filtered by cleanup program, geography, environmental indicators, controls, and cleanup stage. CIMC also provides some web services that integrate these data for others to use in their applications.

Protective effects of buckwheat honey on DNA damage induced by hydroxyl radicals.

PubMed

Zhou, Juan; Li, Peng; Cheng, Ni; Gao, Hui; Wang, Bini; Wei, Yahui; Cao, Wei

2012-08-01

To understand the antioxidant properties of buckwheat honeys, we investigated their antioxidant effects on hydroxyl radical-induced DNA breaks in the non-site-specific and site-specific systems, the physicochemical properties, antioxidant activities (1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical scavenging activity, chelating, and reducing power assays), total phenolic content and individual phenolic acids were also determined. Total phenolic content of buckwheat honeys ranged from 774 to 1694 mg PA/kg, and p-hydroxybenzoic and p-coumaric acids proved to be the main components in buckwheat honeys. All the buckwheat honey samples possess stronger capability to protect DNA in the non-site-specific systems than in the site-specific systems from being damaged by hydroxyl radicals. In the non-site-specific and site-specific system, buckwheat honeys samples prevented ()OH-induced DNA breaks by 21-78% and 5-31% over control value, respectively. Copyright © 2012 Elsevier Ltd. All rights reserved.

Perceptions of the effect of information and communication technology on the quality of care delivered in emergency departments: a cross-site qualitative study.

PubMed

Callen, Joanne; Paoloni, Richard; Li, Julie; Stewart, Michael; Gibson, Kathryn; Georgiou, Andrew; Braithwaite, Jeffrey; Westbrook, Johanna

2013-02-01

We identify and describe emergency physicians' and nurses' perceptions of the effect of an integrated emergency department (ED) information system on the quality of care delivered in the ED. A qualitative study was conducted in 4 urban EDs, with each site using the same ED information system. Participants (n=97) were physicians and nurses with data collected by 69 detailed interviews, 5 focus groups (28 participants), and 26 hours of structured observations. Results revealed new perspectives on how an integrated ED information system was perceived to affect incentives for use, awareness of colleagues' activities, and workflow. A key incentive was related to the positive effect of the ED information system on clinical decisionmaking because of improved and quicker access to patient-specific and knowledge-base information compared with the previous stand-alone ED information system. Synchronous access to patient data was perceived to lead to enhanced awareness by individual physicians and nurses of what others were doing within and outside the ED, which participants claimed contributed to improved care coordination, communication, clinical documentation, and the consultation process. There was difficulty incorporating the use of the ED information system with clinicians' work, particularly in relation to increased task complexity; duplicate documentation, and computer issues related to system usability, hardware, and individuals' computer skills and knowledge. Physicians and nurses perceived that the integrated ED information system contributed to improvements in the delivery of patient care, enabling faster and better-informed decisionmaking and specialty consultations. The challenge of electronic clinical documentation and balancing data entry demands with system benefits necessitates that new methods of data capture, suited to busy clinical environments, be developed. Copyright © 2012. Published by Mosby, Inc.

Active site electrostatics protect genome integrity by blocking abortive hydrolysis during DNA recombination

PubMed Central

Ma, Chien-Hui; Rowley, Paul A; Macieszak, Anna; Guga, Piotr; Jayaram, Makkuni

2009-01-01

Water, acting as a rogue nucleophile, can disrupt transesterification steps of important phosphoryl transfer reactions in DNA and RNA. We have unveiled this risk, and identified safeguards instituted against it, during strand cleavage and joining by the tyrosine site-specific recombinase Flp. Strand joining is threatened by a latent Flp endonuclease activity (type I) towards the 3′-phosphotyrosyl intermediate resulting from strand cleavage. This risk is not alleviated by phosphate electrostatics; neutralizing the negative charge on the scissile phosphate through methylphosphonate (MeP) substitution does not stimulate type I endonuclease. Rather, protection derives from the architecture of the recombination synapse and conformational dynamics within it. Strand cleavage is protected against water by active site electrostatics. Replacement of the catalytic Arg-308 of Flp by alanine, along with MeP substitution, elicits a second Flp endonuclease activity (type II) that directly targets the scissile phosphodiester bond in DNA. MeP substitution, combined with appropriate active site mutations, will be useful in revealing anti-hydrolytic mechanisms engendered by systems that mediate DNA relaxation, DNA transposition, site-specific recombination, telomere resolution, RNA splicing and retrohoming of mobile introns. PMID:19440204

Unexploded Ordnance Characterization And Detection in Muddy Estuarine Environments

NASA Astrophysics Data System (ADS)

Trembanis, A. C.; DuVal, C.

2017-12-01

There is recognized need for better quantitative understanding of the impact of coastal environments on UXO mobility, burial, and detection. Current efforts are underway to address aspects of UXO mobility and detection in sandy coastal areas. However, a significant data gap has been identified regarding UXO in shallow, muddy environments; 139 Formally Used Defense Sites (FUDS), in U.S. tidal waters alone, have been identified as containing muddy sediments. This study works to address this data gap. Using a shallow estuarine site in the Delaware Bay, this study 1) monitors the mobility and behavior of sensor-integrated surrogate munitions in muddy environments using a high-accuracy acoustic positioning system, 2) directly observes surrogate munition response to hydrodynamic forcing through instrumented bottom frame time-lapse hydrodynamic data and sonar imagery, and 3) monitors site changes through repetitive site surveying autonomous underwater vehicle (AUV) using both sonar and magnetometry. Surrogate UXO, modified with acoustic tracking devices and inertial motion units (IMU), are being deployed at a previously characterized muddy estuarine site. The surrogates are being monitored for changes in mobility and burial using the VEMCO positioning system, an off-the-shelf acoustic positioning system that is capable of tracking the position of multiple acoustic tags with accuracies down to 10 cm. Concurrently, time-series acoustic imagery and hydrodynamic sensors are being deployed to characterize UXO response to varied hydrodynamic conditions and compared to site-wide surrogate behavior. A series of repetitive surveys are being conducted using a magnetometer specifically designed for UXO detection on an autonomous underwater vehicle (AUV). Survey results will be compared to long-term acoustic positioning of the surrogate UXO to determine the effectiveness of the magnetometer for efficiently and effectively locating UXO in shallow, muddy environments. Additionally, this study will help inform parameters for UXO mobility and behavior in storms and muddy environments for integration into existing expert system models of UXO burial and mobility.

[Validation of two indices of biological integrity (IBI) for the Angulo River subbasin in Central Mexico].

PubMed

Ramírez-Herrejón, Juan Pablo; Mercado-Silva, Norman; Medina-Nava, Martina; Domínguez-Domínguez, Omar

2012-12-01

Efforts to halt freshwater ecosystem degradation in central Mexico can benefit from using bio-monitoring tools that reflect the condition of their biotic integrity. We analyzed the applicability of two fish-based indices of biotic integrity using data from lotic and lentic systems in the Angulo River subbasin (Lerma-Chapala basin). Both independent data from our own collections during two consecutive years, and existing information detailing the ecological attributes of each species, were used to calculate indices of biological integrity for 16 sites in lotic and lentic habitats. We assessed environmental quality by combining independent evaluations water and habitat quality for each site. We found sites with poor, regular and good biotic integrity. Our study did not find sites with good environmental quality. Fish-based IBI scores were strongly and significantly correlated with scores from independent environmental assessment techniques. IBI scores were adequate at representing environmental conditions in most study sites. These results expand the area where a lotic system fish-based IBI can be used, and constitute an initial validation of a lentic system fish-based IBI. Our results suggest that these bio-monitoring tools can be used in future conservation efforts in freshwater ecosystems in the Middle Lerma Basin.

Centralized Data Management in a Multicountry, Multisite Population-based Study.

PubMed

Rahman, Qazi Sadeq-ur; Islam, Mohammad Shahidul; Hossain, Belal; Hossain, Tanvir; Connor, Nicholas E; Jaman, Md Jahiduj; Rahman, Md Mahmudur; Ahmed, A S M Nawshad Uddin; Ahmed, Imran; Ali, Murtaza; Moin, Syed Mamun Ibne; Mullany, Luke; Saha, Samir K; El Arifeen, Shams

2016-05-01

A centralized data management system was developed for data collection and processing for the Aetiology of Neonatal Infection in South Asia (ANISA) study. ANISA is a longitudinal cohort study involving neonatal infection surveillance and etiology detection in multiple sites in South Asia. The primary goal of designing such a system was to collect and store data from different sites in a standardized way to pool the data for analysis. We designed the data management system centrally and implemented it to enable data entry at individual sites. This system uses validation rules and audit that reduce errors. The study sites employ a dual data entry method to minimize keystroke errors. They upload collected data weekly to a central server via internet to create a pooled central database. Any inconsistent data identified in the central database are flagged and corrected after discussion with the relevant site. The ANISA Data Coordination Centre in Dhaka provides technical support for operations, maintenance and updating the data management system centrally. Password-protected login identifications and audit trails are maintained for the management system to ensure the integrity and safety of stored data. Centralized management of the ANISA database helps to use common data capture forms (DCFs), adapted to site-specific contextual requirements. DCFs and data entry interfaces allow on-site data entry. This reduces the workload as DCFs do not need to be shipped to a single location for entry. It also improves data quality as all collected data from ANISA goes through the same quality check and cleaning process.

Integration Testing of Space Flight Systems

NASA Technical Reports Server (NTRS)

Sowards, Stephanie; Honeycutt, Timothy

2008-01-01

This paper discusses the benefits of conducting multi-system integration testing of space flight elements in lieu of merely shipping and shooting to the launch site and launching. "Ship and shoot" is a philosophy that proposes to transport flight elements directly from the factory to the launch site and begin the mission without further testing. Integration testing, relevant to validation testing in this context, is a risk mitigation effort that builds upon the individual element and system levels of qualification and acceptance tests, greatly improving the confidence of operations in space. The International Space Station Program (ISSP) experience is the focus of most discussions from a historical perspective, while proposed integration testing of the Constellation Program is also discussed. The latter will include Multi-Element Integration Testing (MElT) and Flight Element Integration Testing (FElT).

Overview of DYMCAS, the Y-12 Material Control And Accountability System

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

Alspaugh, D. H.

2001-07-01

This paper gives an overview of DYMCAS, the material control and accountability information system for the Y-12 National Security Complex. A common misconception, even within the DOE community, understates the nature and complexity of material control and accountability (MC and A) systems, likening them to parcel delivery systems tracking packages at various locations or banking systems that account for money, down to the penny. A major point set forth in this paper is that MC and A systems such as DYMCAS can be and often are very complex. Given accountability reporting requirements and the critical and sensitive nature of themore » task, no MC and A system can be simple. The complexity of site-level accountability systems, however, varies dramatically depending on the amounts, kinds, and forms of nuclear materials and the kinds of processing performed at the site. Some accountability systems are tailored to unique and highly complex site-level materials and material processing and, consequently, are highly complex systems. Sites with less complexity require less complex accountability systems, and where processes and practices are the same or similar, sites on the mid-to-low end of the complexity scale can effectively utilize a standard accountability system. In addition to being complex, a unique feature of DYMCAS is its integration with the site production control and manufacturing system. This paper will review the advantages of such integration, as well as related challenges, and make the point that the effectiveness of complex MC and A systems can be significantly enhanced through appropriate systems integration.« less

Comparison of fuel-cell and diesel integrated energy systems and a conventional system for a 500-unit apartment

NASA Technical Reports Server (NTRS)

Simons, S. N.; Maag, W. L.

1978-01-01

The electrical and thermal energy utilization efficiencies of a 500 unit apartment complex are analyzed and compared for each of three energy supply systems. Two on-site integrated energy systems, one powered by diesel engines and the other by phosphoric-acid fuel cells were compared with a conventional system which uses purchased electricity and on-site boilers for heating. All fuels consumed on-site are clean, synthetic fuels (distillate fuel oil or pipeline quality gas) derived from coal. Purchased electricity was generated from coal at a central station utility. The relative energy consumption and economics of the three systems are analyzed and compared.

Specific insertions of zinc finger domains into Gag-Pol yield engineered retroviral vectors with selective integration properties

PubMed Central

Lim, Kwang-il; Klimczak, Ryan; Yu, Julie H.; Schaffer, David V.

2010-01-01

Retroviral vectors offer benefits of efficient delivery and stable gene expression; however, their clinical use raises the concerns of insertional mutagenesis and potential oncogenesis due to genomic integration preferences in transcriptional start sites (TSS). We have shifted the integration preferences of retroviral vectors by generating a library of viral variants with a DNA-binding domain inserted at random positions throughout murine leukemia virus Gag-Pol, then selecting for variants that are viable and exhibit altered integration properties. We found seven permissive zinc finger domain (ZFD) insertion sites throughout Gag-Pol, including within p12, reverse transcriptase, and integrase. Comprehensive genome integration analysis showed that several ZFD insertions yielded retroviral vector variants with shifted integration patterns that did not favor TSS. Furthermore, integration site analysis revealed selective integration for numerous mutants. For example, two retroviral variants with a given ZFD at appropriate positions in Gag-Pol strikingly integrated primarily into four common sites out of 3.1 × 109 possible human genome locations (P = 4.6 × 10-29). Our findings demonstrate that insertion of DNA-binding motifs into multiple locations in Gag-Pol can make considerable progress toward engineering safer retroviral vectors that integrate into a significantly narrowed pool of sites on human genome and overcome the preference for TSS. PMID:20616052

High-Affinity Quasi-Specific Sites in the Genome: How the DNA-Binding Proteins Cope with Them

PubMed Central

Chakrabarti, J.; Chandra, Navin; Raha, Paromita; Roy, Siddhartha

2011-01-01

Many prokaryotic transcription factors home in on one or a few target sites in the presence of a huge number of nonspecific sites. Our analysis of λ-repressor in the Escherichia coli genome based on single basepair substitution experiments shows the presence of hundreds of sites having binding energy within 3 Kcal/mole of the OR1 binding energy, and thousands of sites with binding energy above the nonspecific binding energy. The effect of such sites on DNA-based processes has not been fully explored. The presence of such sites dramatically lowers the occupation probability of the specific site far more than if the genome were composed of nonspecific sites only. Our Brownian dynamics studies show that the presence of quasi-specific sites results in very significant kinetic effects as well. In contrast to λ-repressor, the E. coli genome has orders of magnitude lower quasi-specific sites for GalR, an integral transcription factor, thus causing little competition for the specific site. We propose that GalR and perhaps repressors of the same family have evolved binding modes that lead to much smaller numbers of quasi-specific sites to remove the untoward effects of genomic DNA. PMID:21889449

Techniques for Large-Scale Bacterial Genome Manipulation and Characterization of the Mutants with Respect to In Silico Metabolic Reconstructions.

PubMed

diCenzo, George C; Finan, Turlough M

2018-01-01

The rate at which all genes within a bacterial genome can be identified far exceeds the ability to characterize these genes. To assist in associating genes with cellular functions, a large-scale bacterial genome deletion approach can be employed to rapidly screen tens to thousands of genes for desired phenotypes. Here, we provide a detailed protocol for the generation of deletions of large segments of bacterial genomes that relies on the activity of a site-specific recombinase. In this procedure, two recombinase recognition target sequences are introduced into known positions of a bacterial genome through single cross-over plasmid integration. Subsequent expression of the site-specific recombinase mediates recombination between the two target sequences, resulting in the excision of the intervening region and its loss from the genome. We further illustrate how this deletion system can be readily adapted to function as a large-scale in vivo cloning procedure, in which the region excised from the genome is captured as a replicative plasmid. We next provide a procedure for the metabolic analysis of bacterial large-scale genome deletion mutants using the Biolog Phenotype MicroArray™ system. Finally, a pipeline is described, and a sample Matlab script is provided, for the integration of the obtained data with a draft metabolic reconstruction for the refinement of the reactions and gene-protein-reaction relationships in a metabolic reconstruction.

Expanding the Scope of Site-Specific Recombinases for Genetic and Metabolic Engineering

PubMed Central

Gaj, Thomas; Sirk, Shannon J.; Barbas, Carlos F.

2014-01-01

Site-specific recombinases are tremendously valuable tools for basic research and genetic engineering. By promoting high-fidelity DNA modifications, site-specific recombination systems have empowered researchers with unprecedented control over diverse biological functions, enabling countless insights into cellular structure and function. The rigid target specificities of many sites-specific recombinases, however, have limited their adoption in fields that require highly flexible recognition abilities. As a result, intense effort has been directed toward altering the properties of site-specific recombination systems by protein engineering. Here, we review key developments in the rational design and directed molecular evolution of site-specific recombinases, highlighting the numerous applications of these enzymes across diverse fields of study. PMID:23982993

Assessing your competitors' application of CIM/CIP. [Computer Integrated Manufacturing/Processing

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

King, M.J.; Evans, H.N.

1993-07-01

As part of the authors consulting assignments, they are frequently asked to describe what is best industry practice in the area of computer integrated manufacturing/processing (CIM/CIP). This might be specific to a particular piece, such as advanced controls or a laboratory system. Often it is in response to the enormous publicity given to CIM/CIP--begging the question, Who in the hydrocarbon industry is actually doing it '' Although much of this information is available to consultants, client confidentiality precludes its release. Instead, included is a questionnaire intended to be completed by representatives of manufacturing sites. The data gathered will be analyzedmore » and reported in a future issue. The intent is to give anyone who has completed the questionnaire the opportunity to assess the position of his or her site with respect to the competition. To show how this might work a prototype study was completed. This included an estimate of the advanced control benefits achieved in 68 refineries in Western Europe. So that sites could be compared, these were expressed as a percentage of the maximum economically achievable.« less

Potential of Social Networking Sites for Distance Education Student Engagement

ERIC Educational Resources Information Center

Lester, Jaime; Perini, Michael

2010-01-01

This chapter explores the potential of social networking sites for increasing student engagement for distance education learners. The authors present a modified student engagement model with a focus on the integration of technology, specifically social networking sites for community college distance education learners. The chapter concludes with…

WHAT ARE THE BEST MEANS TO ASSESS SITES AND MOVE TOWARD CLOSURE, USING APPROPRIATE SITE SPECIFIC RISK EVALUATIONS?

EPA Science Inventory

To facilitate evaluation of existing site characterization data, ORD has developed on-line tools and models that integrate data and models into innovative applications. Forty calculators have been developed in four groups: parameter estimators, models, scientific demos and unit ...

Long-Term Environmental Research Programs - Evolving Capacity for Discovery

NASA Astrophysics Data System (ADS)

Swanson, F. J.

2008-12-01

Long-term forestry, watershed, and ecological research sites have become critical, productive nodes for environmental science research and in some cases for work in the social sciences and humanities. The Forest Service's century-old Experimental Forests and Ranges and the National Science Foundation's 28- year-old Long-Term Ecological Research program have been remarkably productive in both basic and applied sciences, including characterization of acid rain and old-growth ecosystems and development of forest, watershed, and range management systems for commercial and other land use objectives. A review of recent developments suggests steps to enhance the function of collections of long-term research sites as interactive science networks. The programs at these sites have evolved greatly, especially over the past few decades, as the questions addressed, disciplines engaged, and degree of science integration have grown. This is well displayed by small, experimental watershed studies, which first were used for applied hydrology studies then more fundamental biogeochemical studies and now examination of complex ecosystem processes; all capitalizing on the legacy of intensive studies and environmental monitoring spanning decades. In very modest ways these collections of initially independent sites have functioned increasingly as integrated research networks addressing inter-site questions by using common experimental designs, being part of a single experiment, and examining long-term data in a common analytical framework. The network aspects include data sharing via publicly-accessible data-harvester systems for climate and streamflow data. The layering of one research or environmental monitoring network upon another facilitates synergies. Changing climate and atmospheric chemistry highlight a need to use these networks as continental-scale observatory systems for assessing the impacts of environmental change on ecological services. To better capitalize on long-term research sites and networks, agencies and universities 1) need to encourage collaboration among sites and between science and land manager communities while 2) maintaining long- term studies and monitoring efforts, and staffing the collaboration in each partner organization, including positions specifically designated as liaisons among the participating communities.

Site-specific protein immobilization in a microfluidic chip channel via an IEF-gelation process.

PubMed

Shi, Mianhong; Peng, Youyuan; Yu, Shaoning; Liu, Baohong; Kong, Jilie

2007-05-01

A novel strategy for site-specific protein immobilization via combining chip IEF with low-temperature sol-gel technology, called IEF-GEL here, in the channel of a modified poly(methyl methacrylate) (PMMA) microfluidic chip is proposed in this work. The IEF-GEL process involves firstly IEF for homogeneously dissolved protein in PBS containing alumina sol and carrier ampholyte with prearranged pH gradient, and then gelation locally for protein encapsulation. The process and feasibility of proposed IEF-GEL were investigated by EOF measurements, fluorescence microscopic photography, Raman spectrum and further demonstrated by glucose oxidase (GOx) reactors integrated with end-column electrochemical detection. Site-controllable immobilization of protein was realized in a 30 mm long microfluidic chip channel by the strategy to create a approximately 1.7 mm concentrated FITC-BSA band, which leads to great improvement of the elute peak shape, accomplished with remarkably increased sensitivity, approximately 20 times higher than that without IEF-GEL treatment to GOx reactors. The kinetic response of GOx after IEF-GEL treatment was also investigated. The proposed system holds the advantages of IEF and low-temperature sol-gel technologies, i.e. concentrating the protein to be focused and retaining the biological activity for the gel-embedded protein, thus realizes site-specific immobilization of low-concentration protein at nL volume level.

Gene-specific cell labeling using MiMIC transposons.

PubMed

Gnerer, Joshua P; Venken, Koen J T; Dierick, Herman A

2015-04-30

Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Implementation of two new resource management information systems in Australia

NASA Astrophysics Data System (ADS)

Kessell, Stephen R.; Good, Roger B.; Hopkins, Angas J. M.

1984-05-01

This paper describes the development and implementation of PREPLAN, A Pristine Environment Planning Language and Simulator, for two conservation areas in Australia, Kosciusko National Park (New South Wales) and Tutanning Nature Reserve (Western Australia). PREPLAN was derived from the North American gradient modeling systems and the Forest Planning Language and Simulator (FORPLAN), but includes unique characteristics not previously available. PREPLAN includes an integrated resource management data base, modules for predicting site-specific vegetation, fuels, animals, fire behavior, and fire effects, and an English language instruction set. PREPLAN was developed specifically to provide available information and understanding of ecosystems to managers in a readily accessible and usable form, and to provide the motivation to conduct additional required research projects. An evaluation of the system's advantages and limitations is presented, and the way the utilization of such systems is improving natural area decision making throughout Australia is discussed.

Clinical experience with a high-performance ATM-connected DICOM archive for cardiology

NASA Astrophysics Data System (ADS)

Solomon, Harry P.

1997-05-01

A system to archive large image sets, such as cardiac cine runs, with near realtime response must address several functional and performance issues, including efficient use of a high performance network connection with standard protocols, an architecture which effectively integrates both short- and long-term mass storage devices, and a flexible data management policy which allows optimization of image distribution and retrieval strategies based on modality and site-specific operational use. Clinical experience with such as archive has allowed evaluation of these systems issues and refinement of a traffic model for cardiac angiography.

Integrating proximal and satellite optical data for the analysis of ecosystem carbon uptake and plant phenology at the European larch Specnet site

NASA Astrophysics Data System (ADS)

Galvagno, Marta; Gamon, John; Cremonese, Edoardo; Garrity, Steven; Huemmrich, K. Fred; Filippa, Gianluca; Morra di Cella, Umberto; Rossini, Micol

2017-04-01

Automated canopy-level optical sampling in tandem with ecosystem-atmosphere flux observations is continuously carried on at a variety of ecosystems through the Specnet network (http://specnet.info/). Specifically, 9 sites within US and Europe were selected since 2015, to investigate the use of novel NDVI and PRI low-cost sensors for the analysis of ecosystem functioning and phenology. Different plant functional types, such as grasslands, deciduous, and evergreen forests belong to the network, here we present specific data from the larch (Larix decidua Mill.) forest Italian site. Three automated NDVI and three automated PRI spectral reflectance sensors (Decagon Devices Inc.) were installed in 2015 on the top of the 20-meters eddy covariance tower, pointing toward the west, north, and east orientations. An additional system, composed by one NDVI and PRI system was installed to monitor the understory component. The objective of this analysis is the comparison between these in-situ inexpensive sensors, independent NDVI and PRI sensors (Skye Instruments) previously installed on the 20-meters tower and satellite-derived NDVI. Both MODIS and Sentinel NDVI data were used for the comparison. Moreover, the newly derived chlorophyll/carotenoid index (CCI, Gamon et al. 2016), computed as the normalized difference between the NDVI red band and PRI 532 nm band, was tested to estimate the seasonal pattern of daily Gross Primary Productivity (GPP) of the larch forest. Results showed that the seasonality of NDVI was comparable among in-situ sensors and satellite data, though orientation-specific differences were observed. Both NDVI and CCI tracked daily GPP, but with different sensitivity to its seasonality. Future analysis will be directed toward a comparison between this site-based results with the other sites within the Specnet network.

Cell-laden hydrogel/titanium microhybrids: Site-specific cell delivery to metallic implants for improved integration.

PubMed

Koenig, Geraldine; Ozcelik, Hayriye; Haesler, Lisa; Cihova, Martina; Ciftci, Sait; Dupret-Bories, Agnes; Debry, Christian; Stelzle, Martin; Lavalle, Philippe; Vrana, Nihal Engin

2016-03-01

Porous titanium implants are widely used in dental, orthopaedic and otorhinolaryngology fields to improve implant integration to host tissue. A possible step further to improve the integration with the host is the incorporation of autologous cells in porous titanium structures via cell-laden hydrogels. Fast gelling hydrogels have advantageous properties for in situ applications such as localisation of specific cells and growth factors at a target area without dispersion. The ability to control the cell types in different regions of an implant is important in applications where the target tissue (i) has structural heterogeneity (multiple cell types with a defined spatial configuration with respect to each other); (ii) has physical property gradients essential for its function (such as in the case of osteochondral tissue transition). Due to their near immediate gelation, such gels can also be used for site-specific modification of porous titanium structures, particularly for implants which would face different tissues at different locations. Herein, we describe a step by step design of a model system: the model cell-laden gel-containing porous titanium implants in the form of titanium microbead/hydrogel (maleimide-dextran or maleimide-PVA based) microhybrids. These systems enable the determination of the effect of titanium presence on gel properties and encapsulated cell behaviour as a miniaturized version of full-scale implants, providing a system compatible with conventional analysis methods. We used a fibroblast/vascular endothelial cell co-cultures as our model system and by utilising single microbeads we have quantified the effect of gel microenvironment (degradability, presence of RGD peptides within gel formulation) on cell behaviour and the effect of the titanium presence on cell behaviour and gel formation. Titanium presence slightly changed gel properties without hindering gel formation or affecting cell viability. Cells showed a preference to move towards the titanium beads and fibroblast proliferation was significantly higher in hybrids compared to gel only controls. The MMP (Matrix Metalloproteinase)-sensitive hydrogels induced sprouting by cells in co-culture configuration which was quantified by fluorescence microscopy, confocal microscopy and qRT-PCR (Quantitative Reverse transcription polymerase chain reaction). When the microhybrid up-scaled to 3D thick structures, cellular localisation in specific areas of the 3D titanium structures was achieved, without decreasing overall cell proliferation compared to titanium only scaffolds. Microhybrids of titanium and hydrogels are useful models for deciding the necessary modifications of metallic implants and they can be used as a modelling system for the study of tissue/titanium implant interactions. This article demonstrates a method to apply cell-laden hydrogels to porous titanium implants and a model of titanium/hydrogel interaction at micro-level using titanium microbeads. The feasibility of site-specific modification of titanium implants with cell-laden microgels has been demonstrated. Use of titanium microbeads in combination with hydrogels with conventional analysis techniques as described in the article can facilitate the characterisation of surface modification of titanium in a relevant model system. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Development of a Comprehensive Database System for Safety Analyst

PubMed Central

Paz, Alexander; Veeramisti, Naveen; Khanal, Indira; Baker, Justin

2015-01-01

This study addressed barriers associated with the use of Safety Analyst, a state-of-the-art tool that has been developed to assist during the entire Traffic Safety Management process but that is not widely used due to a number of challenges as described in this paper. As part of this study, a comprehensive database system and tools to provide data to multiple traffic safety applications, with a focus on Safety Analyst, were developed. A number of data management tools were developed to extract, collect, transform, integrate, and load the data. The system includes consistency-checking capabilities to ensure the adequate insertion and update of data into the database. This system focused on data from roadways, ramps, intersections, and traffic characteristics for Safety Analyst. To test the proposed system and tools, data from Clark County, which is the largest county in Nevada and includes the cities of Las Vegas, Henderson, Boulder City, and North Las Vegas, was used. The database and Safety Analyst together help identify the sites with the potential for safety improvements. Specifically, this study examined the results from two case studies. The first case study, which identified sites having a potential for safety improvements with respect to fatal and all injury crashes, included all roadway elements and used default and calibrated Safety Performance Functions (SPFs). The second case study identified sites having a potential for safety improvements with respect to fatal and all injury crashes, specifically regarding intersections; it used default and calibrated SPFs as well. Conclusions were developed for the calibration of safety performance functions and the classification of site subtypes. Guidelines were provided about the selection of a particular network screening type or performance measure for network screening. PMID:26167531

Expanding the scope of site-specific recombinases for genetic and metabolic engineering.

PubMed

Gaj, Thomas; Sirk, Shannon J; Barbas, Carlos F

2014-01-01

Site-specific recombinases are tremendously valuable tools for basic research and genetic engineering. By promoting high-fidelity DNA modifications, site-specific recombination systems have empowered researchers with unprecedented control over diverse biological functions, enabling countless insights into cellular structure and function. The rigid target specificities of many sites-specific recombinases, however, have limited their adoption in fields that require highly flexible recognition abilities. As a result, intense effort has been directed toward altering the properties of site-specific recombination systems by protein engineering. Here, we review key developments in the rational design and directed molecular evolution of site-specific recombinases, highlighting the numerous applications of these enzymes across diverse fields of study. © 2013 Wiley Periodicals, Inc.

Technology Catalogue. First edition

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

Not Available

1994-02-01

The Department of Energy`s Office of Environmental Restoration and Waste Management (EM) is responsible for remediating its contaminated sites and managing its waste inventory in a safe and efficient manner. EM`s Office of Technology Development (OTD) supports applied research and demonstration efforts to develop and transfer innovative, cost-effective technologies to its site clean-up and waste management programs within EM`s Office of Environmental Restoration and Office of Waste Management. The purpose of the Technology Catalogue is to provide performance data on OTD-developed technologies to scientists and engineers assessing and recommending technical solutions within the Department`s clean-up and waste management programs, asmore » well as to industry, other federal and state agencies, and the academic community. OTD`s applied research and demonstration activities are conducted in programs referred to as Integrated Demonstrations (IDs) and Integrated Programs (IPs). The IDs test and evaluate.systems, consisting of coupled technologies, at specific sites to address generic problems, such as the sensing, treatment, and disposal of buried waste containers. The IPs support applied research activities in specific applications areas, such as in situ remediation, efficient separations processes, and site characterization. The Technology Catalogue is a means for communicating the status. of the development of these innovative technologies. The FY93 Technology Catalogue features technologies successfully demonstrated in the field through IDs and sufficiently mature to be used in the near-term. Technologies from the following IDs are featured in the FY93 Technology Catalogue: Buried Waste ID (Idaho National Engineering Laboratory, Idaho); Mixed Waste Landfill ID (Sandia National Laboratories, New Mexico); Underground Storage Tank ID (Hanford, Washington); Volatile organic compound (VOC) Arid ID (Richland, Washington); and VOC Non-Arid ID (Savannah River Site, South Carolina).« less

Landslides risk mitigation along lifelines

NASA Astrophysics Data System (ADS)

Capparelli, G.; Versace, P.; Artese, G.; Costanzo, S.; Corsonello, P.; Di Massa, G.; Mendicino, G.; Maletta, D.; Leone, S.; Muto, F.; Senatore, A.; Troncone, A.; Conte, E.; Galletta, D.

2012-04-01

The paper describes an integrated, innovative and efficient solution to manage risk issues associated to landslides interfering with infrastructures. The research project was submitted for financial support in the framework of the Multi -regional Operational Programme 2007-13: Research and Competitiveness funded by the Ministry of Research (MIUR) and co-funded by the European Regional Development Fund. The project is aimed to developing and demonstrating an integrated system of monitoring, early warning and mitigation of landslides risk. The final goal is to timely identify potentially dangerous landslides, and to activate all needed impact mitigation measures, including the information delivery. The essential components of the system include monitoring arrays, telecommunication networks and scenario simulation models, assisted by a data acquisition and processing centre, and a traffic control centres. Upon integration, the system will be experimentally validated and demonstrated over ca. 200 km of three highway sections, crossing the regions of Campania, Basilicata, Calabria and Sicily. Progress in the state of art is represented by the developments in the field of environmental monitoring and in the mathematical modeling of landslides and by the development of services for traffic management. The approach to the problem corresponds to a "systemic logics" where each developed component foresees different interchangeable technological solutions to maximize the operational flexibility. The final system may be configured as a simple to complex structure, including different configurations to deal with different scenarios. Specifically, six different monitoring systems will be realized: three "point" systems, made up of a network of locally measuring sensors, and three "area" systems to remotely measure the displacements of large areas. Each network will be fully integrated and connected to a unique data transmission system. Standardized and shared procedures for the identification of risk scenarios will be developed, concerning the surveys to be carried out, the procedures for each type of on-site testing and guidelines and dynamic templates for presentations of results, such as highway risk maps e.g. The setting up of data acquisition and processing centre and traffic control centre are the core of the integrated system. The DAC (data acquisition center, newly designed) will acquire and process data varying in intensity, dimensions, characteristics and information content. The Traffic Control Center (TCC) is meant to integrate the scientific and the management aspects of hydrological risk monitoring and early warning. The overall system is expected to benefit of the development of new, advanced mathematical models on landslide triggers and propagation. Triggering models will be empirical or hydrological, represented by simple empirical relationships, obtained by linking the antecedent rainfall and the landslide time occurrence, and complete models identified through more complex expressions that take into account different components as the specific site conditions, the mechanical, hydraulic and physical properties of soils and slopes, the local seepage conditions and their contribution to soil strength. The industrial partners of the University of Calabria are Autostrade Tech, Strago and TD Group, with the Universities of Firenze and Catania acting research Partners.

Integrated Remote Sensing Modalities for Classification at a Legacy Test Site

NASA Astrophysics Data System (ADS)

Lee, D. J.; Anderson, D.; Craven, J.

2016-12-01

Detecting, locating, and characterizing suspected underground nuclear test sites is of interest to the worldwide nonproliferation monitoring community. Remote sensing provides both cultural and surface geological information over a large search area in a non-intrusive manner. We have characterized a legacy nuclear test site at the Nevada National Security Site (NNSS) using an aerial system based on RGB imagery, light detection and ranging, and hyperspectral imaging. We integrate these different remote sensing modalities to perform pattern recognition and classification tasks on the test site. These tasks include detecting cultural artifacts and exotic materials. We evaluate if the integration of different remote sensing modalities improves classification performance.

Data Integration Tool: From Permafrost Data Translation Research Tool to A Robust Research Application

NASA Astrophysics Data System (ADS)

Wilcox, H.; Schaefer, K. M.; Jafarov, E. E.; Strawhacker, C.; Pulsifer, P. L.; Thurmes, N.

2016-12-01

The United States National Science Foundation funded PermaData project led by the National Snow and Ice Data Center (NSIDC) with a team from the Global Terrestrial Network for Permafrost (GTN-P) aimed to improve permafrost data access and discovery. We developed a Data Integration Tool (DIT) to significantly speed up the time of manual processing needed to translate inconsistent, scattered historical permafrost data into files ready to ingest directly into the GTN-P. We leverage this data to support science research and policy decisions. DIT is a workflow manager that divides data preparation and analysis into a series of steps or operations called widgets. Each widget does a specific operation, such as read, multiply by a constant, sort, plot, and write data. DIT allows the user to select and order the widgets as desired to meet their specific needs. Originally it was written to capture a scientist's personal, iterative, data manipulation and quality control process of visually and programmatically iterating through inconsistent input data, examining it to find problems, adding operations to address the problems, and rerunning until the data could be translated into the GTN-P standard format. Iterative development of this tool led to a Fortran/Python hybrid then, with consideration of users, licensing, version control, packaging, and workflow, to a publically available, robust, usable application. Transitioning to Python allowed the use of open source frameworks for the workflow core and integration with a javascript graphical workflow interface. DIT is targeted to automatically handle 90% of the data processing for field scientists, modelers, and non-discipline scientists. It is available as an open source tool in GitHub packaged for a subset of Mac, Windows, and UNIX systems as a desktop application with a graphical workflow manager. DIT was used to completely translate one dataset (133 sites) that was successfully added to GTN-P, nearly translate three datasets (270 sites), and is scheduled to translate 10 more datasets ( 1000 sites) from the legacy inactive site data holdings of the Frozen Ground Data Center (FGDC). Iterative development has provided the permafrost and wider scientific community with an extendable tool designed specifically for the iterative process of translating unruly data.

Toward the Design of Personalized Continuum Surgical Robots.

PubMed

Morimoto, Tania K; Greer, Joseph D; Hawkes, Elliot W; Hsieh, Michael H; Okamura, Allison M

2018-05-31

Robot-assisted minimally invasive surgical systems enable procedures with reduced pain, recovery time, and scarring compared to traditional surgery. While these improvements benefit a large number of patients, safe access to diseased sites is not always possible for specialized patient groups, including pediatric patients, due to their anatomical differences. We propose a patient-specific design paradigm that leverages the surgeon's expertise to design and fabricate robots based on preoperative medical images. The components of the patient-specific robot design process are a virtual reality design interface enabling the surgeon to design patient-specific tools, 3-D printing of these tools with a biodegradable polyester, and an actuation and control system for deployment. The designed robot is a concentric tube robot, a type of continuum robot constructed from precurved, elastic, nesting tubes. We demonstrate the overall patient-specific design workflow, from preoperative images to physical implementation, for an example clinical scenario: nonlinear renal access to a pediatric kidney. We also measure the system's behavior as it is deployed through real and artificial tissue. System integration and successful benchtop experiments in ex vivo liver and in a phantom patient model demonstrate the feasibility of using a patient-specific design workflow to plan, fabricate, and deploy personalized, flexible continuum robots.

Adeno-associated virus Rep-mediated targeting of integrase-defective retroviral vector DNA circles into human chromosome 19

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

Huang, Shuohao; Kawabe, Yoshinori; Ito, Akira

2012-01-06

Highlights: Black-Right-Pointing-Pointer Adeno-associated virus (AAV) is capable of targeted integration in human cells. Black-Right-Pointing-Pointer Integrase-defective retroviral vector (IDRV) enables a circular DNA delivery. Black-Right-Pointing-Pointer A targeted integration system of IDRV DNA using the AAV integration mechanism. Black-Right-Pointing-Pointer Targeted IDRV integration ameliorates the safety concerns for retroviral vectors. -- Abstract: Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors,more » therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.« less

The UAH GeoIntegrator: A Web Mapping System for On-site Data Insertion and Viewing

NASA Astrophysics Data System (ADS)

He, M.; Hardin, D.; Sever, T.; Irwin, D.

2005-12-01

There is a growing need in the scientific community to combine data colleted in the field with maps, imagery and other layered sources. For example, a biologist, who has collected pollination data during a field study, may want to see his data presented on a regional map. There are many commercial web mapping tools available, but they are expensive, and may require advanced computer knowledge to operate. Researchers from the Information Technology and Systems Center at the University of Alabama in Huntsville are developing a web mapping system that will allow scientists to map their data in an easy way. This system is called the UAH GeoIntegrator. The UAH GeoIntegrator is built on top of three open-source components: the Apache web server, MapServer, and the Chameleon viewer. Chameleon allows developers to customize its map viewer interface by adding widgets. These widgets provide unique functionality focused to the specific needs of the researcher. The UAH GeoIntegrator utilizes a suite of widgets that bring new functionality focused on specific needs, to a typical web map viewer. Specifically, a common input text file format was defined and widgets developed to convert user's field collections into web map layers. These layers can then laid on top of other map layers to produce data products that are versatile, informative and easy to distribute via web services. The UAH GeoIntegrator is being developed as part of the SERVIR project. SERVIR (a Spanish acronym meaning to serve) is part of an international effort to preserve the remaining forested regions of Mesoamerica and to help establish sustainable development in the region. The National Aeronautics and Space Administration along with the World Bank, the United States Agency for International Development and the Central American Commission for Environment and Development are cooperating in this effort. The UAH GeoIntegrator is part of an advanced decision support system that will provide scientists, educators, and policy makers the capabilities needed to monitor and forecast ecological changes, respond to natural disasters, and better understand both natural and human induced effects in Mesoamerica. In this paper, the architecture of the system, data input format, and details of the suite of will be presented.

78 FR 9029 - Nez Perce-Clearwater National Forests; ID; Clear Creek Integrated Restoration Project

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-07

... Nez Perce Forest Plan, and would adopt the Regional soils standard for the Clear Creek Integrated... transportation needs. Site-specifically amend the Nez Perce Forest Plan (1987) to adopt the Region 1 soil standard of 15% for detrimentally compacted, displaced, or puddled soils for the Clear Creek Integrated...

Systems integration of marketable subsystems: A collection of progress reports

NASA Technical Reports Server (NTRS)

1978-01-01

Monthly progress reports are given in the areas of marketable subsystems integration; development, design, and building of site data acquisition subsystems and data processing systems; operation of the solar test facility and a systems analysis.

Integrated Care Planning for Cancer Patients: A Scoping Review

PubMed Central

Arthurs, Erin; Gradin, Sharon; MacKinnon, Marnie; Sussman, Jonathan; Kukreti, Vishal

2017-01-01

Introduction: There has been a growing emphasis on the use of integrated care plans to deliver cancer care. However little is known about how integrated care plans for cancer patients are developed including featured core activities, facilitators for uptake and indicators for assessing impact. Methods: Given limited consensus around what constitutes an integrated care plan for cancer patients, a scoping review was conducted to explore the components of integrated care plans and contextual factors that influence design and uptake. Results: Five types of integrated care plans based on the stage of cancer care: surgical, systemic, survivorship, palliative and comprehensive (involving a transition between stages) are described in current literature. Breast, esophageal and colorectal cancers were common disease sites. Multi-disciplinary teams, patient needs assessment and transitional planning emerged as key features. Provider buy-in and training alongside informational technology support served as important facilitators for plan uptake. Provider-level measurement was considerably less robust compared to patient and system-level indicators. Conclusions: Similarities in design features, components and facilitators across the various types of integrated care plans indicates opportunities to leverage shared features and enable a management lens that spans the trajectory of a patient’s journey rather than a phase-specific silo approach to care. PMID:29588638

NCAR Integrated Sounding System Observations during the SOAS / SAS Field Campaign

NASA Astrophysics Data System (ADS)

Brown, W. O.; Moore, J.

2013-12-01

The National Center for Atmospheric Research (NCAR) Earth Observing Laboratory (EOL) deployed an Integrated Sounding Systems (ISS) for the SOAS (Southern Oxidant and Aerosol Study) field campaign in Alabama in the summer of 2013. The ISS was split between two sites: a former NWS site approximately 1km from the main SOAS chemistry ground site near Centerville AL, and about 20km to the south at the Alabama fish hatchery site approximately 1km from the flux tower site near Marion, AL. At the former-NWS site we launched 106 radiosonde soundings, operated a 915 MHz boundary layer radar wind profiler with RASS (Radio Acoustic Sounding System), ceilometer and various surface meteorological sensors. At the AABC site we operated a Lesosphere WIndcube 200S Doppler lidar and a Metek mini-Doppler sodar. Other NCAR facilities at the AABC site included a 45-m instrumented flux tower. This poster will present a sampling observations made by these instruments, including examples of boundary layer evolution and structure, and summarize the performance of the instrumentation.

Meeting the challenges of installing a mobile robotic system

NASA Technical Reports Server (NTRS)

Decorte, Celeste

1994-01-01

The challenges of integrating a mobile robotic system into an application environment are many. Most problems inherent to installing the mobile robotic system fall into one of three categories: (1) the physical environment - location(s) where, and conditions under which, the mobile robotic system will work; (2) the technological environment - external equipment with which the mobile robotic system will interact; and (3) the human environment - personnel who will operate and interact with the mobile robotic system. The successful integration of a mobile robotic system into these three types of application environment requires more than a good pair of pliers. The tools for this job include: careful planning, accurate measurement data (as-built drawings), complete technical data of systems to be interfaced, sufficient time and attention of key personnel for training on how to operate and program the robot, on-site access during installation, and a thorough understanding and appreciation - by all concerned - of the mobile robotic system's role in the security mission at the site, as well as the machine's capabilities and limitations. Patience, luck, and a sense of humor are also useful tools to keep handy during a mobile robotic system installation. This paper will discuss some specific examples of problems in each of three categories, and explore approaches to solving these problems. The discussion will draw from the author's experience with on-site installations of mobile robotic systems in various applications. Most of the information discussed in this paper has come directly from knowledge learned during installations of Cybermotion's SR2 security robots. A large part of the discussion will apply to any vehicle with a drive system, collision avoidance, and navigation sensors, which is, of course, what makes a vehicle autonomous. And it is with these sensors and a drive system that the installer must become familiar in order to foresee potential trouble areas in the physical, technical, and human environment.

Development tests for the 2.5 megawatt Mod-2 wind turbine generator

NASA Technical Reports Server (NTRS)

Andrews, J. S.; Baskin, J. M.

1982-01-01

The 2.5 megawatt MOD-2 wind turbine generator test program is discussed. The development of the 2.5 megawatt MOD-2 wind turbine generator included an extensive program of testing which encompassed verification of analytical procedures, component development, and integrated system verification. The test program was to assure achievement of the thirty year design operational life of the wind turbine system as well as to minimize costly design modifications which would otherwise have been required during on site system testing. Computer codes were modified, fatigue life of structure and dynamic components were verified, mechanical and electrical component and subsystems were functionally checked and modified where necessary to meet system specifications, and measured dynamic responses of coupled systems confirmed analytical predictions.

Performance Evaluation of Speech Recognition Systems as a Next-Generation Pilot-Vehicle Interface Technology

NASA Technical Reports Server (NTRS)

Arthur, Jarvis J., III; Shelton, Kevin J.; Prinzel, Lawrence J., III; Bailey, Randall E.

2016-01-01

During the flight trials known as Gulfstream-V Synthetic Vision Systems Integrated Technology Evaluation (GV-SITE), a Speech Recognition System (SRS) was used by the evaluation pilots. The SRS system was intended to be an intuitive interface for display control (rather than knobs, buttons, etc.). This paper describes the performance of the current "state of the art" Speech Recognition System (SRS). The commercially available technology was evaluated as an application for possible inclusion in commercial aircraft flight decks as a crew-to-vehicle interface. Specifically, the technology is to be used as an interface from aircrew to the onboard displays, controls, and flight management tasks. A flight test of a SRS as well as a laboratory test was conducted.

System integration of marketable subsystems. [for residential solar heating and cooling

NASA Technical Reports Server (NTRS)

1979-01-01

Progress is reported in the following areas: systems integration of marketable subsystems; development, design, and building of site data acquisition subsystems; development and operation of the central data processing system; operation of the MSFC Solar Test Facility; and systems analysis.

A novel restriction endonuclease GlaI for rapid and highly sensitive detection of DNA methylation coupled with isothermal exponential amplification reaction.

PubMed

Sun, Yueying; Sun, Yuanyuan; Tian, Weimin; Liu, Chenghui; Gao, Kejian; Li, Zhengping

2018-02-07

Sensitive and accurate detection of site-specific DNA methylation is of critical significance for early diagnosis of human diseases, especially cancers. Herein, for the first time we employ a novel methylation-dependent restriction endonuclease GlaI to detect site-specific DNA methylation in a highly specific and sensitive way by coupling with isothermal exponential amplification reaction (EXPAR). GlaI can only cut the methylated target site with excellent selectivity but leave the unmethylated DNA intact. Then the newly exposed end fragments of methylated DNA can trigger EXPAR for highly efficient signal amplification while the intact unmethylated DNA will not initiate EXPAR at all. As such, only the methylated DNA is quantitatively and faithfully reflected by the real-time fluorescence signal of the GlaI-EXPAR system, and the potential false positive interference from unmethylated DNA can be effectively eliminated. Therefore, by integrating the unique features of GlaI for highly specific methylation discrimination and EXPAR for rapid and powerful signal amplification, the elegant GlaI-EXPAR assay allows the direct quantification of methylated DNA with ultrahigh sensitivity and accuracy. The detection limit of methylated DNA target has been pushed down to the aM level and the whole detection process of GlaI-EXPAR can be accomplished within a short time of 2 h. More importantly, ultrahigh specificity is achieved and as low as 0.01% methylated DNA can be clearly identified in the presence of a large excess of unmethylated DNA. This GlaI-EXPAR is also demonstrated to be capable of determining site-specific DNA methylations in real genomic DNA samples. Sharing the distinct advantages of ultrahigh sensitivity, outstanding specificity and facile operation, this new GlaI-EXPAR strategy may provide a robust and reliable platform for the detection of site-specific DNA methylations with low abundances.

Retrotransposon Tf1 is targeted to pol II promoters by transcription activators

PubMed Central

Leem, Young-Eun; Ripmaster, Tracy; Kelly, Felice; Ebina, Hirotaka; Heincelman, Marc; Zhang, Ke; Grewal, Shiv I. S.; Hoffman, Charles S.; Levin, Henry L.

2008-01-01

SUMMARY The LTR-retrotransposon Tf1 preserves the coding capacity of its host Schizosaccharomyces pombe by integrating upstream of open reading frames (ORFs). To determine which features of the target sites were recognized by the transposon, we introduced plasmids containing candidate insertion sites into S. pombe and mapped the positions of integration. We found that Tf1 was targeted specifically to the promoters of pol II transcribed genes. A detailed analysis of integration in plasmids that contained either ade6 or fbp1 revealed insertions occurred in the promoters at positions where transcription factors bound. Further experiments revealed that the activator Atf1p and its binding site were required for directing integration to the promoter of fbp1. An interaction between Tf1 integrase and Atf1p was observed indicating that integration at fbp1 was mediated by the activator bound to its promoter. Surprisingly we found Tf1 contained sequences that activated transcription and these substituted for elements of the ade6 promoter disrupted by integration. PMID:18406330

Retrotransposon Tf1 is targeted to Pol II promoters by transcription activators.

PubMed

Leem, Young-Eun; Ripmaster, Tracy L; Kelly, Felice D; Ebina, Hirotaka; Heincelman, Marc E; Zhang, Ke; Grewal, Shiv I S; Hoffman, Charles S; Levin, Henry L

2008-04-11

The LTR-retrotransposon Tf1 preserves the coding capacity of its host Schizosaccharomyces pombe by integrating upstream of open reading frames (ORFs). To determine which features of the target sites were recognized by the transposon, we introduced plasmids containing candidate insertion sites into S. pombe and mapped the positions of integration. We found that Tf1 was targeted specifically to the promoters of Pol II-transcribed genes. A detailed analysis of integration in plasmids that contained either ade6 or fbp1 revealed insertions occurred in the promoters at positions where transcription factors bound. Further experiments revealed that the activator Atf1p and its binding site were required for directing integration to the promoter of fbp1. An interaction between Tf1 integrase and Atf1p was observed, indicating that integration at fbp1 was mediated by the activator bound to its promoter. Surprisingly, we found Tf1 contained sequences that activated transcription, and these substituted for elements of the ade6 promoter disrupted by integration.

Virus-Clip: a fast and memory-efficient viral integration site detection tool at single-base resolution with annotation capability.

PubMed

Ho, Daniel W H; Sze, Karen M F; Ng, Irene O L

2015-08-28

Viral integration into the human genome upon infection is an important risk factor for various human malignancies. We developed viral integration site detection tool called Virus-Clip, which makes use of information extracted from soft-clipped sequencing reads to identify exact positions of human and virus breakpoints of integration events. With initial read alignment to virus reference genome and streamlined procedures, Virus-Clip delivers a simple, fast and memory-efficient solution to viral integration site detection. Moreover, it can also automatically annotate the integration events with the corresponding affected human genes. Virus-Clip has been verified using whole-transcriptome sequencing data and its detection was validated to have satisfactory sensitivity and specificity. Marked advancement in performance was detected, compared to existing tools. It is applicable to versatile types of data including whole-genome sequencing, whole-transcriptome sequencing, and targeted sequencing. Virus-Clip is available at http://web.hku.hk/~dwhho/Virus-Clip.zip.

Free Energy Landscape of Lipid Interactions with Regulatory Binding Sites on the Transmembrane Domain of the EGF Receptor.

PubMed

Hedger, George; Shorthouse, David; Koldsø, Heidi; Sansom, Mark S P

2016-08-25

Lipid molecules can bind to specific sites on integral membrane proteins, modulating their structure and function. We have undertaken coarse-grained simulations to calculate free energy profiles for glycolipids and phospholipids interacting with modulatory sites on the transmembrane helix dimer of the EGF receptor within a lipid bilayer environment. We identify lipid interaction sites at each end of the transmembrane domain and compute interaction free energy profiles for lipids with these sites. Interaction free energies ranged from ca. -40 to -4 kJ/mol for different lipid species. Those lipids (glycolipid GM3 and phosphoinositide PIP2) known to modulate EGFR function exhibit the strongest binding to interaction sites on the EGFR, and we are able to reproduce the preference for interaction with GM3 over other glycolipids suggested by experiment. Mutation of amino acid residues essential for EGFR function reduce the binding free energy of these key lipid species. The residues interacting with the lipids in the simulations are in agreement with those suggested by experimental (mutational) studies. This approach provides a generalizable tool for characterizing the interactions of lipids that bind to specific sites on integral membrane proteins.

Free Energy Landscape of Lipid Interactions with Regulatory Binding Sites on the Transmembrane Domain of the EGF Receptor

PubMed Central

2016-01-01

Lipid molecules can bind to specific sites on integral membrane proteins, modulating their structure and function. We have undertaken coarse-grained simulations to calculate free energy profiles for glycolipids and phospholipids interacting with modulatory sites on the transmembrane helix dimer of the EGF receptor within a lipid bilayer environment. We identify lipid interaction sites at each end of the transmembrane domain and compute interaction free energy profiles for lipids with these sites. Interaction free energies ranged from ca. −40 to −4 kJ/mol for different lipid species. Those lipids (glycolipid GM3 and phosphoinositide PIP2) known to modulate EGFR function exhibit the strongest binding to interaction sites on the EGFR, and we are able to reproduce the preference for interaction with GM3 over other glycolipids suggested by experiment. Mutation of amino acid residues essential for EGFR function reduce the binding free energy of these key lipid species. The residues interacting with the lipids in the simulations are in agreement with those suggested by experimental (mutational) studies. This approach provides a generalizable tool for characterizing the interactions of lipids that bind to specific sites on integral membrane proteins. PMID:27109430

77 FR 10485 - Environmental Management Site-Specific Advisory Board, Idaho National Laboratory

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-22

... Status EM/National Nuclear Security Administration Integration Ecological Surveys Ground Water Waste Area... and site management in the areas of environmental restoration, waste management, and related... Idaho Cleanup Project (ICP) Workforce Reductions Advanced Mixed Waste Cleanup Project (AMWTP) Workforce...

A multi-landing pad DNA integration platform for mammalian cell engineering

PubMed Central

Gaidukov, Leonid; Wroblewska, Liliana; Teague, Brian; Nelson, Tom; Zhang, Xin; Liu, Yan; Jagtap, Kalpana; Mamo, Selamawit; Tseng, Wen Allen; Lowe, Alexis; Das, Jishnu; Bandara, Kalpanie; Baijuraj, Swetha; Summers, Nevin M; Zhang, Lin; Weiss, Ron

2018-01-01

Abstract Engineering mammalian cell lines that stably express many transgenes requires the precise insertion of large amounts of heterologous DNA into well-characterized genomic loci, but current methods are limited. To facilitate reliable large-scale engineering of CHO cells, we identified 21 novel genomic sites that supported stable long-term expression of transgenes, and then constructed cell lines containing one, two or three ‘landing pad’ recombination sites at selected loci. By using a highly efficient BxB1 recombinase along with different selection markers at each site, we directed recombinase-mediated insertion of heterologous DNA to selected sites, including targeting all three with a single transfection. We used this method to controllably integrate up to nine copies of a monoclonal antibody, representing about 100 kb of heterologous DNA in 21 transcriptional units. Because the integration was targeted to pre-validated loci, recombinant protein expression remained stable for weeks and additional copies of the antibody cassette in the integrated payload resulted in a linear increase in antibody expression. Overall, this multi-copy site-specific integration platform allows for controllable and reproducible insertion of large amounts of DNA into stable genomic sites, which has broad applications for mammalian synthetic biology, recombinant protein production and biomanufacturing. PMID:29617873

Geomembrane barriers using integral fiber optics to monitor barrier integrity

DOEpatents

Staller, G.E.; Wemple, R.P.

1996-10-22

This invention provides a geomembrane or geotextile with embedded optical sensors that are used to monitor the status of containment site barriers. Fiber optic strands are used to form the sensors that can detect and monitor conditions at the sites such as breaches, slope creep, subsidence, leachate levels, fires, and types of materials present or leaking from the site. The strands are integral to the membrane or textile materials. The geosynthetic membrane is deployed at the site in a fashion similar to carpet laying. Edges of the membrane or textile are joined to form a liner and the ends of the membrane or textile become the connection zones for obtaining signals from the sensors. A connection interface with a control system to generate Optical Time Delay Response or other light signals for transmission to the optic fiber strands or sensors and also to receive reflected signals from the sensors is included in the system. Software to interpret the sensor signals can be used in the geosynthetic monitoring system. 6 figs.

Geomembrane barriers using integral fiber optics to monitor barrier integrity

DOEpatents

Staller, George E.; Wemple, Robert P.

1996-01-01

This invention provides a geomembrane or geotextile with embedded optical sensors that are used to monitor the status of containment site barriers. Fiber optic strands are used to form the sensors that can detect and monitor conditions at the sites such as breaches, slope creep, subsidence, leachate levels, fires, and types of materials present or leaking from the site. The strands are integral to the membrane or textile materials. The geosythetic membrane is deployed at the site in a fashion similar to carpet laying. Edges of the membrane or textile are joined to form a liner and the ends of the membrane or textile become the connection zones for obtaining signals from the sensors. A connection interface with a control system to generate Optical Time Delay Response or other light signals for transmission to the optic fiber strands or sensors and also to receive reflected signals from the sensors is included in the system. Software to interpret the sensor signals can be used in the geosythetic monitoring system.

Cellular and molecular mechanisms of HIV-1 integration targeting.

PubMed

Engelman, Alan N; Singh, Parmit K

2018-07-01

Integration is central to HIV-1 replication and helps mold the reservoir of cells that persists in AIDS patients. HIV-1 interacts with specific cellular factors to target integration to interior regions of transcriptionally active genes within gene-dense regions of chromatin. The viral capsid interacts with several proteins that are additionally implicated in virus nuclear import, including cleavage and polyadenylation specificity factor 6, to suppress integration into heterochromatin. The viral integrase protein interacts with transcriptional co-activator lens epithelium-derived growth factor p75 to principally position integration within gene bodies. The integrase additionally senses target DNA distortion and nucleotide sequence to help fine-tune the specific phosphodiester bonds that are cleaved at integration sites. Research into virus-host interactions that underlie HIV-1 integration targeting has aided the development of a novel class of integrase inhibitors and may help to improve the safety of viral-based gene therapy vectors.

Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

PubMed Central

Rollie, Clare; Schneider, Stefanie; Brinkmann, Anna Sophie; Bolt, Edward L; White, Malcolm F

2015-01-01

The adaptive prokaryotic immune system CRISPR-Cas provides RNA-mediated protection from invading genetic elements. The fundamental basis of the system is the ability to capture small pieces of foreign DNA for incorporation into the genome at the CRISPR locus, a process known as Adaptation, which is dependent on the Cas1 and Cas2 proteins. We demonstrate that Cas1 catalyses an efficient trans-esterification reaction on branched DNA substrates, which represents the reverse- or disintegration reaction. Cas1 from both Escherichia coli and Sulfolobus solfataricus display sequence specific activity, with a clear preference for the nucleotides flanking the integration site at the leader-repeat 1 boundary of the CRISPR locus. Cas2 is not required for this activity and does not influence the specificity. This suggests that the inherent sequence specificity of Cas1 is a major determinant of the adaptation process. DOI: http://dx.doi.org/10.7554/eLife.08716.001 PMID:26284603

Lipid Interaction Sites on Channels, Transporters and Receptors: Recent Insights from Molecular Dynamics Simulations

PubMed Central

Hedger, George; Sansom, Mark S. P.

2017-01-01

Lipid molecules are able to selectively interact with specific sites on integral membrane proteins, and modulate their structure and function. Identification and characterisation of these sites is of importance for our understanding of the molecular basis of membrane protein function and stability, and may facilitate the design of lipid-like drug molecules. Molecular dynamics simulations provide a powerful tool for the identification of these sites, complementing advances in membrane protein structural biology and biophysics. We describe recent notable biomolecular simulation studies which have identified lipid interaction sites on a range of different membrane proteins. The sites identified in these simulation studies agree well with those identified by complementary experimental techniques. This demonstrates the power of the molecular dynamics approach in the prediction and characterization of lipid interaction sites on integral membrane proteins. PMID:26946244

Experimental Verification and Integration of a Next Generation Smart Power Management System

NASA Astrophysics Data System (ADS)

Clemmer, Tavis B.

With the increase in energy demand by the residential community in this country and the diminishing fossil fuel resources being used for electric energy production there is a need for a system to efficiently manage power within a residence. The Smart Green Power Node (SGPN) is a next generation energy management system that automates on-site energy production, storage, consumption, and grid usage to yield the most savings for both the utility and the consumer. Such a system automatically manages on-site distributed generation sources such as a PhotoVoltaic (PV) input and battery storage to curtail grid energy usage when the price is high. The SGPN high level control features an advanced modular algorithm that incorporates weather data for projected PV generation, battery health monitoring algorithms, user preferences for load prioritization within the home in case of an outage, Time of Use (ToU) grid power pricing, and status of on-site resources to intelligently schedule and manage power flow between the grid, loads, and the on-site resources. The SGPN has a scalable, modular architecture such that it can be customized for user specific applications. This drove the topology for the SGPN which connects on-site resources at a low voltage DC microbus; a two stage bi-directional inverter/rectifier then couples the AC load and residential grid connect to on-site generation. The SGPN has been designed, built, and is undergoing testing. Hardware test results obtained are consistent with the design goals set and indicate that the SGPN is a viable system with recommended changes and future work.

Information Requirements for Integrating Spatially Discrete, Feature-Based Earth Observations

NASA Astrophysics Data System (ADS)

Horsburgh, J. S.; Aufdenkampe, A. K.; Lehnert, K. A.; Mayorga, E.; Hsu, L.; Song, L.; Zaslavsky, I.; Valentine, D. L.

2014-12-01

Several cyberinfrastructures have emerged for sharing observational data collected at densely sampled and/or highly instrumented field sites. These include the CUAHSI Hydrologic Information System (HIS), the Critical Zone Observatory Integrated Data Management System (CZOData), the Integrated Earth Data Applications (IEDA) and EarthChem system, and the Integrated Ocean Observing System (IOOS). These systems rely on standard data encodings and, in some cases, standard semantics for classes of geoscience data. Their focus is on sharing data on the Internet via web services in domain specific encodings or markup languages. While they have made progress in making data available, it still takes investigators significant effort to discover and access datasets from multiple repositories because of inconsistencies in the way domain systems describe, encode, and share data. Yet, there are many scenarios that require efficient integration of these data types across different domains. For example, understanding a soil profile's geochemical response to extreme weather events requires integration of hydrologic and atmospheric time series with geochemical data from soil samples collected over various depth intervals from soil cores or pits at different positions on a landscape. Integrated access to and analysis of data for such studies are hindered because common characteristics of data, including time, location, provenance, methods, and units are described differently within different systems. Integration requires syntactic and semantic translations that can be manual, error-prone, and lossy. We report information requirements identified as part of our work to define an information model for a broad class of earth science data - i.e., spatially-discrete, feature-based earth observations resulting from in-situ sensors and environmental samples. We sought to answer the question: "What information must accompany observational data for them to be archivable and discoverable within a publication system as well as interpretable once retrieved from such a system for analysis and (re)use?" We also describe development of multiple functional schemas (i.e., physical implementations for data storage, transfer, and archival) for the information model that capture the requirements reported here.

Task 10 - technology development integration. Semi-annual report, April 1--September 30, 1996

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

Hendrikson, J.G.; Daly, D.J.

1997-05-01

The Energy and Environmental Research Center (EERC), in conjunction with the Waste Policy Institute (WPI), will identify and integrate new technologies to meet site-specific environmental management (EM) requirements at contaminated sites appropriate to U.S. Department of Energy (DOE) interests. This paper briefly reports overall progress for three activities: technology management, project management, and technology integration. Work performed over the reporting period has focused on providing logistical and administrative support. In addition, six monthly WPI reports to the EERC are included as appendices. The WPI reports contained detailed information for progress in each activity.

Evaluating Site-Specific and Generic Spatial Models of Aboveground Forest Biomass Based on Landsat Time-Series and LiDAR Strip Samples in the Eastern USA

Treesearch

Ram Deo; Matthew Russell; Grant Domke; Hans-Erik Andersen; Warren Cohen; Christopher Woodall

2017-01-01

Large-area assessment of aboveground tree biomass (AGB) to inform regional or national forest monitoring programs can be efficiently carried out by combining remotely sensed data and field sample measurements through a generic statistical model, in contrast to site-specific models. We integrated forest inventory plot data with spatial predictors from Landsat time-...

The λ Integrase Site-specific Recombination Pathway

PubMed Central

LANDY, ARTHUR

2017-01-01

The site-specific recombinase encoded by bacteriophage λ (Int) is responsible for integrating and excising the viral chromosome into and out of the chromosome of its Escherichia coli host. Int carries out a reaction that is highly directional, tightly regulated, and depends upon an ensemble of accessory DNA bending proteins acting on 240 bp of DNA encoding 16 protein binding sites. This additional complexity enables two pathways, integrative and excisive recombination, whose opposite, and effectively irreversible, directions are dictated by different physiological and environmental signals. Int recombinase is a heterobivalent DNA binding protein and each of the four Int protomers, within a multiprotein 400 kDa recombinogenic complex, is thought to bind and, with the aid of DNA bending proteins, bridge one arm- and one core-type DNA site. In the 12 years since the publication of the last review focused solely on the λ site-specific recombination pathway in Mobile DNA II, there has been a great deal of progress in elucidating the molecular details of this pathway. The most dramatic advances in our understanding of the reaction have been in the area of X-ray crystallography where protein-DNA structures have now been determined for of all of the DNA-protein interfaces driving the Int pathway. Building on this foundation of structures, it has been possible to derive models for the assembly of components that determine the regulatory apparatus in the P-arm, and for the overall architectures that define excisive and integrative recombinogenic complexes. The most fundamental additional mechanistic insights derive from the application of hexapeptide inhibitors and single molecule kinetics. PMID:26104711

Construction and Characterization of an in-vivo Linear Covalently Closed DNA Vector Production System

PubMed Central

2012-01-01

Background While safer than their viral counterparts, conventional non-viral gene delivery DNA vectors offer a limited safety profile. They often result in the delivery of unwanted prokaryotic sequences, antibiotic resistance genes, and the bacterial origins of replication to the target, which may lead to the stimulation of unwanted immunological responses due to their chimeric DNA composition. Such vectors may also impart the potential for chromosomal integration, thus potentiating oncogenesis. We sought to engineer an in vivo system for the quick and simple production of safer DNA vector alternatives that were devoid of non-transgene bacterial sequences and would lethally disrupt the host chromosome in the event of an unwanted vector integration event. Results We constructed a parent eukaryotic expression vector possessing a specialized manufactured multi-target site called “Super Sequence”, and engineered E. coli cells (R-cell) that conditionally produce phage-derived recombinase Tel (PY54), TelN (N15), or Cre (P1). Passage of the parent plasmid vector through R-cells under optimized conditions, resulted in rapid, efficient, and one step in vivo generation of mini lcc—linear covalently closed (Tel/TelN-cell), or mini ccc—circular covalently closed (Cre-cell), DNA constructs, separated from the backbone plasmid DNA. Site-specific integration of lcc plasmids into the host chromosome resulted in chromosomal disruption and 105 fold lower viability than that seen with the ccc counterpart. Conclusion We offer a high efficiency mini DNA vector production system that confers simple, rapid and scalable in vivo production of mini lcc DNA vectors that possess all the benefits of “minicircle” DNA vectors and virtually eliminate the potential for undesirable vector integration events. PMID:23216697

Construction and characterization of an in-vivo linear covalently closed DNA vector production system.

PubMed

Nafissi, Nafiseh; Slavcev, Roderick

2012-12-06

While safer than their viral counterparts, conventional non-viral gene delivery DNA vectors offer a limited safety profile. They often result in the delivery of unwanted prokaryotic sequences, antibiotic resistance genes, and the bacterial origins of replication to the target, which may lead to the stimulation of unwanted immunological responses due to their chimeric DNA composition. Such vectors may also impart the potential for chromosomal integration, thus potentiating oncogenesis. We sought to engineer an in vivo system for the quick and simple production of safer DNA vector alternatives that were devoid of non-transgene bacterial sequences and would lethally disrupt the host chromosome in the event of an unwanted vector integration event. We constructed a parent eukaryotic expression vector possessing a specialized manufactured multi-target site called "Super Sequence", and engineered E. coli cells (R-cell) that conditionally produce phage-derived recombinase Tel (PY54), TelN (N15), or Cre (P1). Passage of the parent plasmid vector through R-cells under optimized conditions, resulted in rapid, efficient, and one step in vivo generation of mini lcc--linear covalently closed (Tel/TelN-cell), or mini ccc--circular covalently closed (Cre-cell), DNA constructs, separated from the backbone plasmid DNA. Site-specific integration of lcc plasmids into the host chromosome resulted in chromosomal disruption and 10(5) fold lower viability than that seen with the ccc counterpart. We offer a high efficiency mini DNA vector production system that confers simple, rapid and scalable in vivo production of mini lcc DNA vectors that possess all the benefits of "minicircle" DNA vectors and virtually eliminate the potential for undesirable vector integration events.

Systems Biophysics of Gene Expression

PubMed Central

Vilar, Jose M.G.; Saiz, Leonor

2013-01-01

Gene expression is a process central to any form of life. It involves multiple temporal and functional scales that extend from specific protein-DNA interactions to the coordinated regulation of multiple genes in response to intracellular and extracellular changes. This diversity in scales poses fundamental challenges to the use of traditional approaches to fully understand even the simplest gene expression systems. Recent advances in computational systems biophysics have provided promising avenues to reliably integrate the molecular detail of biophysical process into the system behavior. Here, we review recent advances in the description of gene regulation as a system of biophysical processes that extend from specific protein-DNA interactions to the combinatorial assembly of nucleoprotein complexes. There is now basic mechanistic understanding on how promoters controlled by multiple, local and distal, DNA binding sites for transcription factors can actively control transcriptional noise, cell-to-cell variability, and other properties of gene regulation, including precision and flexibility of the transcriptional responses. PMID:23790365

Results from a Field Trial of the Radio Frequency Based Cylinder Accountability and Tracking System at the Global Nuclear Fuel Americas Fuel Fabrication Facility

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

Fitzgerald, Peter; Laughter, Mark D; Martyn, Rose

The Cylinder Accountability and Tracking System (CATS) is a tool designed for use by the International Atomic Energy Agency (IAEA) to improve overall inspector efficiency through real-time unattended monitoring of cylinder movements, site specific rules-based event detection, and the capability to integrate many types of monitoring technologies. The system is based on the tracking of cylinder movements using (radio frequency) RF tags, and the collection of data, such as accountability weights, that can be associated with the cylinders. This presentation will cover the installation and evaluation of the CATS at the Global Nuclear Fuels (GNF) fuel fabrication facility in Wilmington,more » NC. This system was installed to evaluate its safeguards applicability, operational durability under operating conditions, and overall performance. An overview of the system design and elements specific to the GNF deployment will be presented along with lessons learned from the installation process and results from the field trial.« less

Anatomical parameterization for volumetric meshing of the liver

NASA Astrophysics Data System (ADS)

Vera, Sergio; González Ballester, Miguel A.; Gil, Debora

2014-03-01

A coordinate system describing the interior of organs is a powerful tool for a systematic localization of injured tissue. If the same coordinate values are assigned to specific anatomical landmarks, the coordinate system allows integration of data across different medical image modalities. Harmonic mappings have been used to produce parametric coordinate systems over the surface of anatomical shapes, given their flexibility to set values at specific locations through boundary conditions. However, most of the existing implementations in medical imaging restrict to either anatomical surfaces, or the depth coordinate with boundary conditions is given at sites of limited geometric diversity. In this paper we present a method for anatomical volumetric parameterization that extends current harmonic parameterizations to the interior anatomy using information provided by the volume medial surface. We have applied the methodology to define a common reference system for the liver shape and functional anatomy. This reference system sets a solid base for creating anatomical models of the patient's liver, and allows comparing livers from several patients in a common framework of reference.

Integrated Logistics Support approach: concept for the new big projects: E-ELT, SKA, CTA

NASA Astrophysics Data System (ADS)

Marchiori, G.; Rampini, F.; Formentin, F.

2014-08-01

The Integrated Logistic Support is a process supporting strategies and optimizing activities for a correct project management and system engineering development. From the design & engineering of complex technical systems, to the erection on site, acceptance and after-sales service, EIE GROUP covers all aspects of the Integrated Logistics Support (ILS) process that includes: costing process centered around the life cycle cost and Level of Repair Analyses; engineering process which influences the design via means of reliability, modularization, etc.; technical publishing process based on international specifications; ordering administration process for supply support. Through the ILS, EIE GROUP plans and directs the identification and development of logistics support and system requirements for its products, with the goal of creating systems that last longer and require less support, thereby reducing costs and increasing return on investments. ILS therefore, addresses these aspects of supportability not only during acquisition, but also throughout the operational life cycle of the system. The impact of the ILS is often measured in terms of metrics such as reliability, availability, maintainability and testability (RAMT), and System Safety (RAMS). Example of the criteria and approach adopted by EIE GROUP during the design, manufacturing and test of the ALMA European Antennas and during the design phase of the E-ELT telescope and Dome are presented.

The integrated contaminant elution and tracer test toolkit, ICET3, for improved characterization of mass transfer, attenuation, and mass removal

NASA Astrophysics Data System (ADS)

Brusseau, Mark L.; Guo, Zhilin

2018-01-01

It is evident based on historical data that groundwater contaminant plumes persist at many sites, requiring costly long-term management. High-resolution site-characterization methods are needed to support accurate risk assessments and to select, design, and operate effective remediation operations. Most subsurface characterization methods are generally limited in their ability to provide unambiguous, real-time delineation of specific processes affecting mass-transfer, transformation, and mass removal, and accurate estimation of associated rates. An integrated contaminant elution and tracer test toolkit, comprising a set of local-scale groundwater extraction-and injection tests, was developed to ameliorate the primary limitations associated with standard characterization methods. The test employs extended groundwater extraction to stress the system and induce hydraulic and concentration gradients. Clean water can be injected, which removes the resident aqueous contaminant mass present in the higher-permeability zones and isolates the test zone from the surrounding plume. This ensures that the concentrations and fluxes measured within the isolated area are directly and predominantly influenced by the local mass-transfer and transformation processes controlling mass removal. A suite of standard and novel tracers can be used to delineate specific mass-transfer and attenuation processes that are active at a given site, and to quantify the associated mass-transfer and transformation rates. The conceptual basis for the test is first presented, followed by an illustrative application based on simulations produced with a 3-D mathematical model and a brief case study application.

Application of aerial photography to water-related programs in Michigan

NASA Technical Reports Server (NTRS)

Enslin, W. R.; Hill-Rowley, R.; Tilmann, S. E.

1977-01-01

The paper describes the use of aerial photography and information system technology in the provision of information required for the effective operation of three water-related programs in Michigan. Potential mosquito breeding sites were identified from specially acquired low altitude 70 mm color photography for the City of Lansing Vector Control Area. A comprehensive inventory of surface water sources and potential access sites was prepared to assist fire departments in Antrim County with fire truck water-recharge operations. Remotely-sensed land cover/use data for Windsor Township, Eaton County were integrated with other resource data into a computer-based information system for regional water quality studies. Eleven thematic maps specifically focussed on landscape features affecting non-point water pollution and waste disposal were generated from analyses of a four-hectare grid-based data file containing land cover/use, soils, topographic and geologic (well-log) data.

Cryogenic reactant storage for lunar base regenerative fuel cells

NASA Technical Reports Server (NTRS)

Kohout, Lisa L.

1989-01-01

There are major advantages to be gained by integrating a cryogenic reactant storage system with a hydrogen-oxygen regenerative fuel cell (RFC) to provide on-site electrical power during the lunar night. Although applicable to any power system using hydrogen-oxygen RFC's for energy storage, cryogenic reactant storage offers a significant benefit whenever the sun/shade cycle and energy storage period approach hundreds of hours. For solar power installations on the moon, cryogenic reactant storage reduces overall specific mass and meteoroid vulnerability of the system. In addition, it offers synergistic benefits to on-site users, such as availability of primary fuel cell reactants for surface rover vehicles and cryogenic propellants for OTV's. The integration involves processing and storing the RFC reactant streams as cryogenic liquids rather than pressurized gases, so that reactant containment (tankage per unit mass of reactants) can be greatly reduced. Hydrogen-oxygen alkaline RFC's, GaAs photovoltaic (PV) arrays, and space cryogenic processing/refrigeration technologies are assumed to be available for the conceptual system design. Advantages are demonstrated by comparing the characteristics of two power system concepts: a conventional lunar surface PV/RFC power system using pressurized gas storage in SOA filament wound pressure vessels and, that same system with gas liquefaction and storage replacing the pressurized storage. Comparisons are made at 20 and 250 kWe. Although cryogenic storage adds a processing plant (drying and liquefaction) to the system plus 30 percent more solar array to provide processing power, the approximate order of magnitude reduction in tankage mass, confirmed by this analysis, results in a reduction in overall total system mass of approximately 50 percent.

Long-term increase in mVEGF164 in mouse hindlimb muscle mediated by phage phiC31 integrase after nonviral DNA delivery.

PubMed

Portlock, Joylette L; Keravala, Annahita; Bertoni, Carmen; Lee, Solomon; Rando, Thomas A; Calos, Michele P

2006-08-01

Peripheral vascular disease (PVD), characterized by insufficient blood supply to extremities, can be a devastating illness. Although many gene therapy strategies for PVD using vascular endothelial growth factor (VEGF) have resulted in increased blood vessel formation, the vessels are often impermanent and regress after therapy, probably because of the short-lived VEGF expression mediated by gene therapy vectors (14 days or less). phiC31 integrase is a recombinase originally isolated from a bacteriophage of Streptomyces. This integrase performs efficient chromosomal integration of plasmid DNA into mammalian genomes that results in long-term transgene expression. In this study, gene transfer was achieved by intramuscular injection of VEGF and integrase plasmid DNAs into the tibialis anterior muscle in the mouse hindlimb, followed by electroporation of the muscle with needle electrodes. We observed VEGF levels significantly above background 40 days after injection in animals that received phiC31 integrase and the VEGF plasmid. Site-specific integration of plasmid DNA in the chromosomes of muscle tissue was verified by polymerase chain reaction at a common integration site. These results suggest the possible utility of the phiC31 integrase system to treat ischemic disease.

Develop and test fuel cell powered on-site integrated total energy system

NASA Technical Reports Server (NTRS)

1983-01-01

Test results are given for a 5 kW stack and initial results for an integrated, grid connected system operating from methanol fuel. Site selection criteria are presented for future demonstration of a 50 or 100 kW OS/IES. Preliminary results are also given with approximate internal rates of return to the building owner. Progress in development and construction of a 50 kW modular methanol/steam reformer is reported.

Analysis of proviral integration in human mammary epithelial cell lines immortalized by retroviral infection with a temperature-sensitive SV40 T-antigen construct.

PubMed

Stamps, A C; Davies, S C; Burman, J; O'Hare, M J

1994-06-15

A panel of eight conditionally immortal lines derived by infection of human breast epithelial cells with an amphotropic retrovirus transducing a ts mutant of SV40 large T-antigen was analyzed with respect to individual retroviral integration patterns. Each line contained multiple integration sites which were clonal and stable over extended passage. Similar integration patterns were observed between individual lines arising separately from the same stock of pre-immortal cells, suggesting a common progenitor. Retroviral integration analysis of pre-immortal cells at different stages of pre-crisis growth showed changes indicative of a progressive transition from polyclonality to clonality as the cells approached crisis. Each of the immortal lines contained a sub-set of the integration sites of their pre-immortal progenitors, with individual combinations and copy numbers of sites. Since all the cell lines appeared to originate from single foci in separate flasks, it is likely that each set arose from a common clone of pre-immortal cells as the result of separate genetic events. There was no evidence from this analysis to suggest that specific integration sites played any part either in the selection of pre-crisis clones or in the subsequent establishment of immortal lines.

Intelligent building system for airport

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

Ancevic, M.

1997-11-01

The Munich airport uses a state-of-the-art intelligent building management system to control systems such as HVAC, runway lights, baggage handling, etc. Planning the new Munich II international airport provided a unique opportunity to use the latest state-of-the-art technical systems, while integrating their control through a single intelligent building management system. Opened in 1992, the airport is Germany`s second-largest airport after Frankfurt. The airport is staffed by 16,000 employees and can handle 17 million passengers a year. The sprawling site encompasses more than 120 buildings. The airport`s distributed control system is specifically designed to optimize the complex`s unique range of functions,more » while providing a high degree of comfort, convenience and safety for airport visitors. With the capacity to control 200,000 points, this system controls more than 112,000 points and integrates 13 major subsystems from nine different vendors. It provides convenient, accessible control of everything including the complex`s power plant, HVAC Control, the terminal`s people-moving functions, interior lighting controls, runway lights, baggage forwarding systems, elevators, and boarding bridges. The airport was named 1993 intelligent building of the year by the Intelligent Buildings Institute Foundation. Its building management system is a striking example of the degree to which a building complex`s functions can be integrated for greater operational control and efficiency.« less

Potential interoperability problems facing multi-site radiation oncology centers in The Netherlands

NASA Astrophysics Data System (ADS)

Scheurleer, J.; Koken, Ph; Wessel, R.

2014-03-01

Aim: To identify potential interoperability problems facing multi-site Radiation Oncology (RO) departments in the Netherlands and solutions for unambiguous multi-system workflows. Specific challenges confronting the RO department of VUmc (RO-VUmc), which is soon to open a satellite department, were characterized. Methods: A nationwide questionnaire survey was conducted to identify possible interoperability problems and solutions. Further detailed information was obtained by in-depth interviews at 3 Dutch RO institutes that already operate in more than one site. Results: The survey had a 100% response rate (n=21). Altogether 95 interoperability problems were described. Most reported problems were on a strategic and semantic level. The majority were DICOM(-RT) and HL7 related (n=65), primarily between treatment planning and verification systems or between departmental and hospital systems. Seven were identified as being relevant for RO-VUmc. Departments have overcome interoperability problems with their own, or with tailor-made vendor solutions. There was little knowledge about or utilization of solutions developed by Integrating the Healthcare Enterprise Radiation Oncology (IHE-RO). Conclusions: Although interoperability problems are still common, solutions have been identified. Awareness of IHE-RO needs to be raised. No major new interoperability problems are predicted as RO-VUmc develops into a multi-site department.

Linking Course Web Sites to Library Collections and Services

ERIC Educational Resources Information Center

Rieger, Oya Y.; Horne, Angela K.; Revels, Ira

2004-01-01

A five-month research study at Cornell University Library (CUL) confirmed the strategic importance of a library presence in faculty-created course Web sites. It sparked specific recommendations to support the seamless integration of the CUL digital library within the virtual learning environments created by faculty.

A network-based training environment: a medical image processing paradigm.

PubMed

Costaridou, L; Panayiotakis, G; Sakellaropoulos, P; Cavouras, D; Dimopoulos, J

1998-01-01

The capability of interactive multimedia and Internet technologies is investigated with respect to the implementation of a distance learning environment. The system is built according to a client-server architecture, based on the Internet infrastructure, composed of server nodes conceptually modelled as WWW sites. Sites are implemented by customization of available components. The environment integrates network-delivered interactive multimedia courses, network-based tutoring, SIG support, information databases of professional interest, as well as course and tutoring management. This capability has been demonstrated by means of an implemented system, validated with digital image processing content, specifically image enhancement. Image enhancement methods are theoretically described and applied to mammograms. Emphasis is given to the interactive presentation of the effects of algorithm parameters on images. The system end-user access depends on available bandwidth, so high-speed access can be achieved via LAN or local ISDN connections. Network based training offers new means of improved access and sharing of learning resources and expertise, as promising supplements in training.

Demonstration of an Integrated Compliance Model for Predicting Copper Fate and Effects in DoD Harbors

DTIC Science & Technology

2008-11-01

seawater that does not include the natural ingredients that buffer the toxic effects of contaminants. As such, federal WQC could be overprotective ...regulation was overprotective (Earley et al., 2007). Implementation of a site-specific WQS in both cases could reduce the likelihood of TMDL actions...for site-specific factors that regulate bioavailability and toxicity, and thus are often overprotective (Seligman and Zirino, 1998; Zirino and

Integrating natural and social sciences to manage sustainably vectors of change in the marine environment: Dogger Bank transnational case study

NASA Astrophysics Data System (ADS)

Burdon, Daryl; Boyes, Suzanne J.; Elliott, Michael; Smyth, Katie; Atkins, Jonathan P.; Barnes, Richard A.; Wurzel, Rüdiger K.

2018-02-01

The management of marine resources is a complex process driven by the dynamics of the natural system and the influence of stakeholders including policy-makers. An integration of natural and social sciences research is required by policy-makers to better understand, and manage sustainably, natural changes and anthropogenic activities within particular marine systems. Given the uncertain development of activities in the marine environment, future scenarios assessments can be used to investigate whether marine policy measures are robust and sustainable. This paper develops an interdisciplinary framework, which incorporates future scenarios assessments, and identifies four main types of evaluation needed to integrate natural and social sciences research to support the integrated management of the marine environment: environmental policy and governance assessments; ecosystem services, indicators and valuation; modelling tools for management evaluations, and risk assessment and risk management. The importance of stakeholder engagement within each evaluation method is highlighted. The paper focuses on the transnational spatial marine management of the Dogger Bank, in the central North Sea, a site which is very important ecologically, economically and politically. Current management practices are reviewed, and research tools to support future management decisions are applied and discussed in relation to two main vectors of change affecting the Dogger Bank, namely commercial fisheries and offshore wind farm developments, and in relation to the need for nature conservation. The input of local knowledge through stakeholder engagement is highlighted as a necessary requirement to produce site-specific policy recommendations for the future management of the Dogger Bank. We present wider policy recommendations to integrate natural and social sciences in a global marine context.

INVESTIGATING UNCERTAINTY AND SENSITIVITY IN INTEGRATED MULTIMEDIA ENVIRONMENTAL MODELS: TOOLS FOR 3MRA

EPA Science Inventory

Sufficiently elucidating uncertainty and sensitivity structures in environmental models can be a difficult task, even for low-order, single-media constructs driven by a unique set of site-specific data. The ensuing challenge of examining ever more complex, integrated, higher-ord...

Site planning and integration fiscal year 1999 multi-year work plan (MYWP) update for WBS 1.8.2.1

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

SCHULTZ, E.A.

The primary mission of the Site Planning and Integration (SP and I) project is to assist Fluor Daniel Project Direction to ensure that all work performed under the Project Hanford Management Contract (PHMC) is adequately planned, executed, controlled, and that performance is measured and reported in an integrated fashion. Furthermore, SP and I is responsible for the development, implementation, and management of systems and processes that integrate technical, schedule, and cost baselines for PHMC work.

Impact of care management processes and integration of care on blood pressure control in diabetes.

PubMed

Wong, Ken; Boulanger, Luke; Smalarz, Amy; Wu, Ning; Fraser, Kimberly; Wogen, Jenifer

2013-02-27

Fragmentation within health care systems may negatively impact the quality of chronic disease patient care. We sought to evaluate the relationship between care management processes (CMP), integration of services, and blood pressure (BP) control among diabetic patients. Retrospective chart reviews were performed for a random sample of adult diabetic hypertensive patients (n = 2,162) from 28 physician organizations in the United States (US). A modified version of the Physician Practice Connection Readiness Survey (PPC-RS) was completed by the chief medical officer at each site. The PPC-RS measured health system organization, delivery system redesign, decision support, clinical information systems, and self-management support, and an integration scale measured structure, functions, and financial risk. Correlations between PPC and integration scores and BP outcomes were assessed using Spearman correlation coefficients. Approximately 39.9% of diabetic patients had controlled BP. Mean total PPC score across sites was 55, with highest mean scores for health system organization (81), followed by design support (60), clinical information systems (57), self-management support (39), and delivery system redesign (39). Mean integration score was 46 (SD 27, range 4-93), and means of subscores were 64 for structure, 33 for financial risk, and 42 for function. Clinical information systems subscore was correlated with uncontrolled BP (r = -0.38, p < 0.05), while association with total PPC score was strong but not significant at p < 0.05 (r = -0.32). Total integration score and the structure subscore were significantly correlated with BP control (r = 0.38, p < 0.05, and r = 0.49, p < 0.01). This study suggests that CMP and service integration may be associated with better outcomes in diabetes, though results were mixed and limited by a small number of participating sites. Primary care implementation of integrated electronic medical records may have a beneficial effect on patient outcomes for diabetes and other chronic diseases.

Report on all ARRA Funded Technical Work

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

None, None

2013-10-05

The main focus of this American Recovery and Reinvestment Act of 2009 (ARRA) funded project was to design an energy efficient carbon capture and storage (CCS) process using the Recipients membrane system for H{sub 2} separation and CO{sub 2} capture. In the ARRA-funded project, the Recipient accelerated development and scale-up of ongoing hydrogen membrane technology research and development (R&D). Specifically, this project focused on accelerating the current R&D work scope of the base program-funded project, involving lab scale tests, detail design of a 250 lb/day H{sub 2} process development unit (PDU), and scale-up of membrane tube and coating manufacturing. Thismore » project scope included the site selection and a Front End Engineering Design (FEED) study of a nominally 4 to 10 ton-per-day (TPD) Pre-Commercial Module (PCM) hydrogen separation membrane system. Process models and techno-economic analysis were updated to include studies on integration of this technology into an Integrated Gasification Combined Cycle (IGCC) power generation system with CCS.« less

Systems and methods for knowledge discovery in spatial data

DOEpatents

Obradovic, Zoran; Fiez, Timothy E.; Vucetic, Slobodan; Lazarevic, Aleksandar; Pokrajac, Dragoljub; Hoskinson, Reed L.

2005-03-08

Systems and methods are provided for knowledge discovery in spatial data as well as to systems and methods for optimizing recipes used in spatial environments such as may be found in precision agriculture. A spatial data analysis and modeling module is provided which allows users to interactively and flexibly analyze and mine spatial data. The spatial data analysis and modeling module applies spatial data mining algorithms through a number of steps. The data loading and generation module obtains or generates spatial data and allows for basic partitioning. The inspection module provides basic statistical analysis. The preprocessing module smoothes and cleans the data and allows for basic manipulation of the data. The partitioning module provides for more advanced data partitioning. The prediction module applies regression and classification algorithms on the spatial data. The integration module enhances prediction methods by combining and integrating models. The recommendation module provides the user with site-specific recommendations as to how to optimize a recipe for a spatial environment such as a fertilizer recipe for an agricultural field.

End of FY10 report - used fuel disposition technical bases and lessons learned : legal and regulatory framework for high-level waste disposition in the United States.

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

Weiner, Ruth F.; Blink, James A.; Rechard, Robert Paul

This report examines the current policy, legal, and regulatory framework pertaining to used nuclear fuel and high level waste management in the United States. The goal is to identify potential changes that if made could add flexibility and possibly improve the chances of successfully implementing technical aspects of a nuclear waste policy. Experience suggests that the regulatory framework should be established prior to initiating future repository development. Concerning specifics of the regulatory framework, reasonable expectation as the standard of proof was successfully implemented and could be retained in the future; yet, the current classification system for radioactive waste, including hazardousmore » constituents, warrants reexamination. Whether or not consideration of multiple sites are considered simultaneously in the future, inclusion of mechanisms such as deliberate use of performance assessment to manage site characterization would be wise. Because of experience gained here and abroad, diversity of geologic media is not particularly necessary as a criterion in site selection guidelines for multiple sites. Stepwise development of the repository program that includes flexibility also warrants serious consideration. Furthermore, integration of the waste management system from storage, transportation, and disposition, should be examined and would be facilitated by integration of the legal and regulatory framework. Finally, in order to enhance acceptability of future repository development, the national policy should be cognizant of those policy and technical attributes that enhance initial acceptance, and those policy and technical attributes that maintain and broaden credibility.« less

Homeologous plastid DNA transformation in tobacco is mediated by multiple recombination events.

PubMed Central

Kavanagh, T A; Thanh, N D; Lao, N T; McGrath, N; Peter, S O; Horváth, E M; Dix, P J; Medgyesy, P

1999-01-01

Efficient plastid transformation has been achieved in Nicotiana tabacum using cloned plastid DNA of Solanum nigrum carrying mutations conferring spectinomycin and streptomycin resistance. The use of the incompletely homologous (homeologous) Solanum plastid DNA as donor resulted in a Nicotiana plastid transformation frequency comparable with that of other experiments where completely homologous plastid DNA was introduced. Physical mapping and nucleotide sequence analysis of the targeted plastid DNA region in the transformants demonstrated efficient site-specific integration of the 7.8-kb Solanum plastid DNA and the exclusion of the vector DNA. The integration of the cloned Solanum plastid DNA into the Nicotiana plastid genome involved multiple recombination events as revealed by the presence of discontinuous tracts of Solanum-specific sequences that were interspersed between Nicotiana-specific markers. Marked position effects resulted in very frequent cointegration of the nonselected peripheral donor markers located adjacent to the vector DNA. Data presented here on the efficiency and features of homeologous plastid DNA recombination are consistent with the existence of an active RecA-mediated, but a diminished mismatch, recombination/repair system in higher-plant plastids. PMID:10388829

Tailoring Care to Vulnerable Populations by Incorporating Social Determinants of Health: the Veterans Health Administration's "Homeless Patient Aligned Care Team" Program.

PubMed

O'Toole, Thomas P; Johnson, Erin E; Aiello, Riccardo; Kane, Vincent; Pape, Lisa

2016-03-31

Although the clinical consequences of homelessness are well described, less is known about the role for health care systems in improving clinical and social outcomes for the homeless. We described the national implementation of a "homeless medical home" initiative in the Veterans Health Administration (VHA) and correlated patient health outcomes with characteristics of high-performing sites. We conducted an observational study of 33 VHA facilities with homeless medical homes and patient- aligned care teams that served more than 14,000 patients. We correlated site-specific health care performance data for the 3,543 homeless veterans enrolled in the program from October 2013 through March 2014, including those receiving ambulatory or acute health care services during the 6 months prior to enrollment in our study and 6 months post-enrollment with corresponding survey data on the Homeless Patient Aligned Care Team (H-PACT) program implementation. We defined high performance as high rates of ambulatory care and reduced use of acute care services. More than 96% of VHA patients enrolled in these programs were concurrently receiving VHA homeless services. Of the 33 sites studied, 82% provided hygiene care (on-site showers, hygiene kits, and laundry), 76% provided transportation, and 55% had an on-site clothes pantry; 42% had a food pantry and provided on-site meals or other food assistance. Six-month patterns of acute-care use pre-enrollment and post-enrollment for 3,543 consecutively enrolled patients showed a 19.0% reduction in emergency department use and a 34.7% reduction in hospitalizations. Three features were significantly associated with high performance: 1) higher staffing ratios than other sites, 1) integration of social supports and social services into clinical care, and 3) outreach to and integration with community agencies. Integrating social determinants of health into clinical care can be effective for high-risk homeless veterans.

Tailoring Care to Vulnerable Populations by Incorporating Social Determinants of Health: the Veterans Health Administration’s “Homeless Patient Aligned Care Team” Program

PubMed Central

Johnson, Erin E.; Aiello, Riccardo; Kane, Vincent; Pape, Lisa

2016-01-01

Introduction Although the clinical consequences of homelessness are well described, less is known about the role for health care systems in improving clinical and social outcomes for the homeless. We described the national implementation of a “homeless medical home” initiative in the Veterans Health Administration (VHA) and correlated patient health outcomes with characteristics of high-performing sites. Methods We conducted an observational study of 33 VHA facilities with homeless medical homes and patient- aligned care teams that served more than 14,000 patients. We correlated site-specific health care performance data for the 3,543 homeless veterans enrolled in the program from October 2013 through March 2014, including those receiving ambulatory or acute health care services during the 6 months prior to enrollment in our study and 6 months post-enrollment with corresponding survey data on the Homeless Patient Aligned Care Team (H-PACT) program implementation. We defined high performance as high rates of ambulatory care and reduced use of acute care services. Results More than 96% of VHA patients enrolled in these programs were concurrently receiving VHA homeless services. Of the 33 sites studied, 82% provided hygiene care (on-site showers, hygiene kits, and laundry), 76% provided transportation, and 55% had an on-site clothes pantry; 42% had a food pantry and provided on-site meals or other food assistance. Six-month patterns of acute-care use pre-enrollment and post-enrollment for 3,543 consecutively enrolled patients showed a 19.0% reduction in emergency department use and a 34.7% reduction in hospitalizations. Three features were significantly associated with high performance: 1) higher staffing ratios than other sites, 1) integration of social supports and social services into clinical care, and 3) outreach to and integration with community agencies. Conclusion Integrating social determinants of health into clinical care can be effective for high-risk homeless veterans. PMID:27032987

Fuel cell on-site integrated energy system parametric analysis of a residential complex

NASA Technical Reports Server (NTRS)

Simons, S. N.

1977-01-01

A parametric energy-use analysis was performed for a large apartment complex served by a fuel cell on-site integrated energy system (OS/IES). The variables parameterized include operating characteristics for four phosphoric acid fuel cells, eight OS/IES energy recovery systems, and four climatic locations. The annual fuel consumption for selected parametric combinations are presented and a breakeven economic analysis is presented for one parametric combination. The results show fuel cell electrical efficiency and system component choice have the greatest effect on annual fuel consumption; fuel cell thermal efficiency and geographic location have less of an effect.

The Drupal Environmental Information Management System Provides Standardization, Flexibility and a Platform for Collaboration

NASA Astrophysics Data System (ADS)

Gries, C.; Vanderbilt, K.; Reid, D.; Melendez-Colom, E.; San Gil, I.

2013-12-01

Over the last five years several Long-Term Ecological Research (LTER) sites have collaboratively developed a standardized yet flexible approach to ecological information management based on the open source Drupal content management system. These LTER sites adopted a common data model for basic metadata necessary to describe data sets, but also used for site management and web presence. Drupal core functionality provides web forms for easy management of information stored in this data model. Custom Drupal extensions were developed to generate XML files conforming to the Ecological Metadata Language (EML) for contribution to the LTER Network Information System (NIS) and other data archives. Each LTER site then took advantage of the flexibility Drupal provides to develop its unique web presence, choosing different themes and adding additional content to the websites. By nature, information presented is highly interlinked which can easily be modeled in Drupal entities and is further supported by a sophisticated tagging system (Fig. 1). Therefore, it is possible to provide the visitor with many different entry points to the site specific information presented. For example, publications and datasets may be grouped for each scientist, for each research project, for each major research theme at the site, making the information presented more accessible for different visitors. Experience gained during the early years was recently used to launch a complete re-write for upgrading to Drupal 7. LTER sites from multiple academic institutions pooled resources in order to partner with professional Drupal developers. Highlights of the new developments are streamlined data entry, improved EML output and integrity, support of IM workflows, a faceted data set search, a highly configurable data exploration tool with intelligent filtering and data download, and, for the mobile age, a responsive web design theme. Seven custom modules and a specific installation profile were developed involving many other community contributed modules, all with an upgrade to Drupal 8 in mind. The collaborative development of the Drupal Ecological Information Management System (DEIMS) has resulted in a product that is standards-based but flexible enough to meet individual site needs. It is available at the Drupal.org website for other small research stations or labs to use, extend and improve according to the open source philosophy. Figure 1: Overview of DEIMS components and interactions

System requirement specification for the IH-10 Integrated Corridor Management System (ICMS) in San Antonio, Texas

DOT National Transportation Integrated Search

2008-03-31

This Requirements Specification Document (RSD) was developed under the project titled TransGuide Integrated Corridor Management Stage 1 as part of the United States Department of Transportation (USDOT) Integrated Corridor Management (ICM) p...

Science Objectives and Design of the European Seas Observatory NETwork (ESONET)

NASA Astrophysics Data System (ADS)

Ruhl, H.; Géli, L.; Karstensen, J.; Colaço, A.; Lampitt, R.; Greinert, J.; Phannkuche, O.; Auffret, Y.

2009-04-01

The needs for a network of ocean observing systems cross many applied and research areas of earth and marine science. Many of the science areas that can be examined using such systems have direct impacts on societal health and well being and our understanding of ocean function in a shifting climate. The European Seas Observatory NETwork (ESONET) Network of Excellence has been evaluating ocean observatory design requirements, data management needs, standardization and interoperability concerns, social implications, outreach and education, as well as financial and legal aspects of developing such a system. ESONET has great potential to address a growing set of Earth science questions that require a broad and integrated network of ocean and seafloor observations. ESONET activities are also importantly integrating researchers in the European Community, as well as internationally. There is now wide recognition that research addressing science questions of international priority, such as understanding the potential impacts of climate change or geohazards like earthquakes and tsunamis should be conducted in a framework that can address questions across adequate temporal and spatial scales. We will present the relevant science priorities in the four interconnected fields of geoscience, physical oceanography, biogeochemistry, and marine ecology, and some of the practical ways in which these questions can be addressed using ESONET. Several key questions persist that will require comprehensive interdisciplinary approaches including: How can monitoring of factors such as seismic activity, fluid pore chemistry and pressure, improve seismic, slope failure, and tsunami warning? To what extent do seabed processes influence ocean physics, biogeochemistry, and marine ecosystems? How are physical and biogeochemical processes that occur at differing scales related? What aspects of physical oceanography and biogeochemical cycling will be most sensitive to climate change? What will the important feedbacks of potential ecological change be on biogeochemical cycles? What are the factors that control the distribution and abundance of marine life and what will the influence of anthropogenic change be? We will outline a set of science objectives and observation parameters to be collected at all ESONET sites, as well as a set of rather specific objectives and thus parameters that might only be measured at some sites. We will also present the preliminary module specifications now being considered by ESONET. In a practical sense the observatory design has been divided into those that will be included in a so called ‘generic' module and those that will be part of science-specific modules. Outlining preliminary module specifications is required to move forward with studies of observatory design and operation. These specifications are importantly provisional and can be updated as science needs and feasibility change. A functional cleavage not only comes between aspects that are considered generic or specific, but also the settings in which those systems will be used. For example, some modules will be on the seabed and some will be moored in the water column. In order to address many of the questions posed above ESONET users will require other supporting data from other programs from local to international levels. Examples of these other data sources include satellite oceanographic data, climatic data, air-sea interface data, and the known distribution and abundances of marine fauna. Thus the connection of ESONET to other programs is integral to its success. The development of ESONET provides a substantial opportunity for ocean science to evolve in Europe. Furthermore, ESONET and several other developing ocean observatory programs are integrating into larger science frameworks including the Global Earth Observation System of Systems (GEOSS) and Global Monitoring of Environment and Security (GMES) programs. It is only in a greater integrated framework that the full potential of the component systems will be realized.

Grand challenges in bioengineered nanorobotics for cancer therapy.

PubMed

Lenaghan, Scott C; Wang, Yongzhong; Xi, Ning; Fukuda, Toshio; Tarn, Tzyhjong; Hamel, William R; Zhang, Mingjun

2013-03-01

One of the grand challenges currently facing engineering, life sciences, and medicine is the development of fully functional nanorobots capable of sensing, decision making, and actuation. These nanorobots may aid in cancer therapy, site-specific drug delivery, circulating diagnostics, advanced surgery, and tissue repair. In this paper, we will discuss, from a bioinspired perspective, the challenges currently facing nanorobotics, including core design, propulsion and power generation, sensing, actuation, control, decision making, and system integration. Using strategies inspired from microorganisms, we will discuss a potential bioengineered nanorobot for cancer therapy.

Restoring tropical forests on bauxite mined lands: lessons from the Brazilian Amazon

Treesearch

John A. Parrotta; Oliver H. Knowles

2001-01-01

Restoring self-sustaining tropical forest ecosystems on surface mined sites is a formidable challenge that requires the integration of proven reclamation techniques and reforestation strategies appropriate to specific site conditions, including landscape biodiversity patterns. Restorationists working in most tropical settings are usually hampered by lack of basic...

Self-Assembled Smart Nanocarriers for Targeted Drug Delivery.

PubMed

Cui, Wei; Li, Junbai; Decher, Gero

2016-02-10

Nanostructured drug-carrier systems promise numerous benefits for drug delivery. They can be engineered to precisely control drug-release rates or to target specific sites within the body with a specific amount of therapeutic agent. However, to achieve the best therapeutic effects, the systems should be designed for carrying the optimum amount of a drug to the desired target where it should be released at the optimum rate for a specified time. Despite numerous attempts, fulfilling all of these requirements in a synergistic way remains a huge challenge. The trend in drug delivery is consequently directed toward integrated multifunctional carrier systems, providing selective recognition in combination with sustained or triggered release. Capsules as vesicular systems enable drugs to be confined for controlled release. Furthermore, carriers modified with recognition groups can enhance the capability of encapsulated drug efficacy. Here, recent advances are reviewed regarding designing and preparing assembled capsules with targeting ligands or size controllable for selective recognition in drug delivery. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Final Environmental Assessment for the Integration and Developmental Testing of High Power Microwave Systems at Edwards Air Force Base

DTIC Science & Technology

2006-10-16

spawning sites, feeding sites, 20 seasonal wetlands or drylands, water quality or quantity, host species or plant pollinators , geological 21...rufus]). 12 These areas also contain relatively large areas of sensitive plants . 13 3.9.2.9 Significant Ecological Areas found within the R-2515 and...for complete aircraft, avionics systems, and integrated 21 airframe weapons support. Thus, HPM testing would continue the evolution of the primary

SXT/R391 Integrative and Conjugative Elements (ICEs) Encode a Novel 'Trap-Door' Strategy for Mobile Element Escape.

PubMed

Ryan, Michael P; Armshaw, Patricia; Pembroke, J Tony

2016-01-01

Integrative conjugative elements (ICEs) are a class of bacterial mobile elements that have the ability to mediate their own integration, excision, and transfer from one host genome to another by a mechanism of site-specific recombination, self-circularisation, and conjugative transfer. Members of the SXT/R391 ICE family of enterobacterial mobile genetic elements display an unusual UV-inducible sensitization function which results in stress induced killing of bacterial cells harboring the ICE. This sensitization has been shown to be associated with a stress induced overexpression of a mobile element encoded conjugative transfer gene, orf43, a traV homolog. This results in cell lysis and release of a circular form of the ICE. Induction of this novel system may allow transfer of an ICE, enhancing its survival potential under conditions not conducive to conjugative transfer.

Impact of Co-Site Interference on L/C-Band Spectrum for UAS Control and Non-Payload Communications

NASA Technical Reports Server (NTRS)

Kerczewski, Robert J.; Bishop, William D.; Hoder, Douglas J.; Shalkhauser, Kurt A.; Wilson, Jeffrey D.

2015-01-01

In order to provide for the safe integration of unmanned aircraft systems into the National Airspace System, the control and non-payload communications (CNPC) link connecting the ground-based pilot with the unmanned aircraft must be highly reliable. A specific requirement is that it must operate using aviation safety radiofrequency spectrum. The 2012 World Radiocommunication Conference (WRC-12) provided a potentially suitable allocation for LOS CNPC spectrum in C-Band at 5030-5091 MHz band which, when combined with a previous allocation in L-Band (960-1164 MHz) may satisfy the LOS spectrum requirement and provide for high reliability through dual-band redundancy. However, the LBand spectrum hosts a number of aeronautical navigation systems which require high-power transmitters on-board the aircraft. These high-power transmitters co-located with sensitive CNPC receivers operating in the same frequency band have the potential to create co-site interference, reducing the performance of the CNPC receivers and ultimately reducing the usability of the L-Band for CNPC. This paper examines the potential for co-site interference, as highlighted in recent flight tests, and discusses the impact on the UAS CNPC spectrum availability and requirements for further testing and analysis.

Model-Based Systems

NASA Technical Reports Server (NTRS)

Frisch, Harold P.

2007-01-01

Engineers, who design systems using text specification documents, focus their work upon the completed system to meet Performance, time and budget goals. Consistency and integrity is difficult to maintain within text documents for a single complex system and more difficult to maintain as several systems are combined into higher-level systems, are maintained over decades, and evolve technically and in performance through updates. This system design approach frequently results in major changes during the system integration and test phase, and in time and budget overruns. Engineers who build system specification documents within a model-based systems environment go a step further and aggregate all of the data. They interrelate all of the data to insure consistency and integrity. After the model is constructed, the various system specification documents are prepared, all from the same database. The consistency and integrity of the model is assured, therefore the consistency and integrity of the various specification documents is insured. This article attempts to define model-based systems relative to such an environment. The intent is to expose the complexity of the enabling problem by outlining what is needed, why it is needed and how needs are being addressed by international standards writing teams.

06. Facilitating Collection of Research and Quality Data in Integrative Medicine Clinical Settings: Views From Academic, Health System and Private Clinics

PubMed Central

Dolor, Rowena; Victorson, David; Amoils, Steve

2013-01-01

Focus Areas: Integrative Approaches to Care The purpose of this panel discussion is to share successful efforts from a practice-based research network (PBRN) including ten integrative medicine clinics. The BraveNet PBRN includes integrative medicine clinics with academic health centers, large health systems, and a stand-alone private practice clinic. While clinical care is prioritized across all of these centers, introducing research into clinical sites oriented to providing care poses challenges that vary by clinic environment. We will highlight some of the unique issues encountered when trying to standardize data collection in sites practicing a patient-centered, whole-systems approach to healing as well as the solutions used to overcome these issues. We will present some operational solutions and data collected from the PBRN's ongoing data registry, entitled PRIMIER. The panel will engage attendees in a dialogue centering on potential for future analyses of existing results, ideas for possible upcoming studies, and creative ways to expand the PBRN data registry to include additional sites that may have expertise and interest in participating.

Model-data integration for developing the Cropland Carbon Monitoring System (CCMS)

NASA Astrophysics Data System (ADS)

Jones, C. D.; Bandaru, V.; Pnvr, K.; Jin, H.; Reddy, A.; Sahajpal, R.; Sedano, F.; Skakun, S.; Wagle, P.; Gowda, P. H.; Hurtt, G. C.; Izaurralde, R. C.

2017-12-01

The Cropland Carbon Monitoring System (CCMS) has been initiated to improve regional estimates of carbon fluxes from croplands in the conterminous United States through integration of terrestrial ecosystem modeling, use of remote-sensing products and publically available datasets, and development of improved landscape and management databases. In order to develop these improved carbon flux estimates, experimental datasets are essential for evaluating the skill of estimates, characterizing the uncertainty of these estimates, characterizing parameter sensitivities, and calibrating specific modeling components. Experiments were sought that included flux tower measurement of CO2 fluxes under production of major agronomic crops. Currently data has been collected from 17 experiments comprising 117 site-years from 12 unique locations. Calibration of terrestrial ecosystem model parameters using available crop productivity and net ecosystem exchange (NEE) measurements resulted in improvements in RMSE of NEE predictions of between 3.78% to 7.67%, while improvements in RMSE for yield ranged from -1.85% to 14.79%. Model sensitivities were dominated by parameters related to leaf area index (LAI) and spring growth, demonstrating considerable capacity for model improvement through development and integration of remote-sensing products. Subsequent analyses will assess the impact of such integrated approaches on skill of cropland carbon flux estimates.

Future Interagency Range and Spaceport Technologies (FIRST) Formulation Products: 1. Transformational Spaceport and Range Concept of Operations. 2. F.I.R.S.T. Business Case Analysis

NASA Technical Reports Server (NTRS)

2005-01-01

The Baseline Report captures range and spaceport capabilities at five sites: KSC, CCAFS, VAFB, Wallops, and Kodiak. The Baseline depicts a future state that relies on existing technology, planned upgrades, and straight-line recapitalization at these sites projected through 2030. The report presents an inventory of current spaceport and range capabilities at these five sites. The baseline is the first part of analyzing a business case for a set of capabilities designed to transform U.S. ground and space launch operations toward a single, integrated national "system" of space transportation systems. The second part of the business case compares current capabilities with technologies needed to support the integrated national "system". The final part, a return on investment analysis, identifies the technologies that best lead to the integrated national system and reduce recurring costs..Numerous data sources were used to define and describe the baseline spaceport and range by identifying major systems and elements and describing capabilities, limitations, and capabilities

Control of Recombination Directionality by the Listeria Phage A118 Protein Gp44 and the Coiled-Coil Motif of Its Serine Integrase.

PubMed

Mandali, Sridhar; Gupta, Kushol; Dawson, Anthony R; Van Duyne, Gregory D; Johnson, Reid C

2017-06-01

The serine integrase of phage A118 catalyzes integrative recombination between attP on the phage and a specific attB locus on the chromosome of Listeria monocytogenes , but it is unable to promote excisive recombination between the hybrid attL and attR sites found on the integrated prophage without assistance by a recombination directionality factor (RDF). We have identified and characterized the phage-encoded RDF Gp44, which activates the A118 integrase for excision and inhibits integration. Gp44 binds to the C-terminal DNA binding domain of integrase, and we have localized the primary binding site to be within the mobile coiled-coil (CC) motif but distinct from the distal tip of the CC that is required for recombination. This interaction is sufficient to inhibit integration, but a second interaction involving the N-terminal end of Gp44 is also required to activate excision. We provide evidence that these two contacts modulate the trajectory of the CC motifs as they extend out from the integrase core in a manner dependent upon the identities of the four att sites. Our results support a model whereby Gp44 shapes the Int-bound complexes to control which att sites can synapse and recombine. IMPORTANCE Serine integrases mediate directional recombination between bacteriophage and bacterial chromosomes. These highly regulated site-specific recombination reactions are integral to the life cycle of temperate phage and, in the case of Listeria monocytogenes lysogenized by A118 family phage, are an essential virulence determinant. Serine integrases are also utilized as tools for genetic engineering and synthetic biology because of their exquisite unidirectional control of the DNA exchange reaction. Here, we identify and characterize the recombination directionality factor (RDF) that activates excision and inhibits integration reactions by the phage A118 integrase. We provide evidence that the A118 RDF binds to and modulates the trajectory of the long coiled-coil motif that extends from the large carboxyl-terminal DNA binding domain and is postulated to control the early steps of recombination site synapsis. Copyright © 2017 American Society for Microbiology.

Integrated primary care: an inclusive three-world view through process metrics and empirical discrimination.

PubMed

Miller, Benjamin F; Mendenhall, Tai J; Malik, Alan D

2009-03-01

Integrating behavioral health services within the primary care setting drives higher levels of collaborative care, and is proving to be an essential part of the solution for our struggling American healthcare system. However, justification for implementing and sustaining integrated and collaborative care has shown to be a formidable task. In an attempt to move beyond conflicting terminology found in the literature, we delineate terms and suggest a standardized nomenclature. Further, we maintain that addressing the three principal worlds of healthcare (clinical, operational, financial) is requisite in making sense of the spectrum of available implementations and ultimately transitioning collaborative care into the mainstream. Using a model that deconstructs process metrics into factors/barriers and generalizes behavioral health provider roles into major categories provides a framework to empirically discriminate between implementations across specific settings. This approach offers practical guidelines for care sites implementing integrated and collaborative care and defines a research framework to produce the evidence required for the aforementioned clinical, operational and financial worlds of this important movement.

Obsidian in the Casas Grandes world: Procurement, exchange, and interaction in Chihuahua, Mexico, CE 1200-1450

DOE PAGES

Dolan, Sean Gregory; Whalen, Michael E.; Minnis, Paul E.; ...

2017-01-09

The Casas Grandes area in Chihuahua, Mexico and the site of Paquime was the center of one the largest regional systems in the U.S. Southwest and Mexican Northwest (SW/NW) during the Medio period (CE 1200-1450). People participated in local and distant exchange networks with groups in the SW/NW, Mesoamerica, and West Mexico. Our knowledge of which obsidian sources people used in Casas Grandes is limited, despite how obsidian could have derived from many different places. We examine how the use of specific obsidian sources may relate to broader political and economic relationships within the Casas Grandes regional system and itsmore » association with the Mimbres and Animas regions of the SW/NW. We sourced 116 artifacts using EDXRF spectrometry from four sites that neighbor Paquime. Results demonstrate people used obsidian from Chihuahua, Sonora, and New Mexico. Furthermore, there were varying levels of social interaction and regional integration because there is diversity in source use at the site level, and Casas Grandes were more connected to the Animas region than Mimbres regarding obsidian resource procurement.« less

Obsidian in the Casas Grandes world: Procurement, exchange, and interaction in Chihuahua, Mexico, CE 1200-1450

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

Dolan, Sean Gregory; Whalen, Michael E.; Minnis, Paul E.

The Casas Grandes area in Chihuahua, Mexico and the site of Paquime was the center of one the largest regional systems in the U.S. Southwest and Mexican Northwest (SW/NW) during the Medio period (CE 1200-1450). People participated in local and distant exchange networks with groups in the SW/NW, Mesoamerica, and West Mexico. Our knowledge of which obsidian sources people used in Casas Grandes is limited, despite how obsidian could have derived from many different places. We examine how the use of specific obsidian sources may relate to broader political and economic relationships within the Casas Grandes regional system and itsmore » association with the Mimbres and Animas regions of the SW/NW. We sourced 116 artifacts using EDXRF spectrometry from four sites that neighbor Paquime. Results demonstrate people used obsidian from Chihuahua, Sonora, and New Mexico. Furthermore, there were varying levels of social interaction and regional integration because there is diversity in source use at the site level, and Casas Grandes were more connected to the Animas region than Mimbres regarding obsidian resource procurement.« less

40 CFR 63.681 - Definitions.

Code of Federal Regulations, 2013 CFR

2013-07-01

... features permanently integrated into the design of the unit. Emission point means an individual tank... system is not a drain and collection system that is designed and operated for the sole purpose of..., or transfer system used to manage off-site material. Off-site material service means any time when a...

40 CFR 63.681 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... features permanently integrated into the design of the unit. Emission point means an individual tank... system is not a drain and collection system that is designed and operated for the sole purpose of..., or transfer system used to manage off-site material. Off-site material service means any time when a...

Vaccine adverse events in a safety net healthcare system and a managed care organization.

PubMed

Narwaney, Komal J; Breslin, Kristin; Ross, Colleen A; Shoup, Jo Ann; Wain, Kris F; Weintraub, Eric S; McNeil, Michael M; Hambidge, Simon J

2017-03-01

The Institute of Medicine, in a 2013 report, recommended that the Vaccine Safety Datalink (VSD) expand collaborations to include more diversity in the study population. Kaiser Permanente Colorado (KPCO), an established VSD site, partnered with Denver Health (DH), an integrated safety net healthcare system, to demonstrate the feasibility of integrating DH data within the VSD. Prior to incorporating the data, we examined the identification of specific vaccine associated adverse events (VAEs) in these two distinct healthcare systems. We conducted retrospective cohort analyses within KPCO and DH to compare select VAEs between the two populations. We examined the following associations between January 1, 2004 and December 31, 2013: Measles, Mumps, and Rubella (MMR) vaccine and febrile seizures in children 2years and younger, intussusception after rotavirus vaccine in infants 4-34weeks, syncope after adolescent vaccines (Tetanus, Diphtheria, acellular Pertussis; Meningococcal and Human Papillomavirus) in adolescents 13-17years and medically attended local reactions after pneumococcal polysaccharide (PPSV23) vaccine in adults 65years and older. Both sites used similar data procurement methods and chart review processes. For seizures after MMR vaccine (KPCO - 3.15vs. DH - 2.97/10,000 doses) and syncope after all adolescent vaccines (KPCO - 3.0vs. DH - 2.37/10,000 doses), the chart confirmed rates were comparable at the two sites. However, for medically attended local reactions after PPSV23, there were differences in chart confirmed rates between the sites (KPCO - 31.65vs. DH - 14.90/10,000 doses). For intussusception after rotavirus vaccine, the number of cases was too low to make a valid comparison (KPCO - 0vs. DH - 0.13/10,000 doses). We demonstrated that data on important targeted VAEs can be captured at DH and rates appear similar to those at KPCO. Work is ongoing on the optimal approach to assimilate DH data as a potential safety net healthcare system in the VSD. Copyright © 2017 Elsevier Ltd. All rights reserved.

Site-specific nutrient management systems

USDA-ARS?s Scientific Manuscript database

Site-specific nutrient management systems were created to manage for spatial and temporal variability in biophysical factors that determine the availability and demand of crop nutrients. These systems differ among geographical regions in the information utilized and way they operate to accomplish th...

Bioavailability and Environmental Regulation - Integrating a Fate & Effects Model with a Biotic Ligand Model for Copper in Seawater

NASA Astrophysics Data System (ADS)

Rivera, I.; Chadwick, B.; Rosen, G.; Wang, P. F.; Paquin, P.; Santore, R.; Ryan, A.

2015-12-01

Understanding the bioavailability of metals in the aquatic environment is important for defining appropriate regulatory constraints. A failure to recognize the importance of bioavailability factors on metal toxicity can result in criteria that are over- or under-protective. USEPA addresses the tendency of the national Water Quality Criterion (WQC) for regulation of copper in marine waters to underestimate the natural attenuation of copper toxicity in harbors by the application of site-specific Water Quality Standards (WQS). Which provides the level of protection intended by the WQC, and establishes realistic regulatory objectives. However, development of site-specific WQS involves a long-term effort, and does not account for temporal variation. The toxicity model seawater-Biotic Ligand Model (BLM) was developed and integrated with the existing Curvilinear Hydrodynamics in 3 Dimensions (CH3D) transport & fate model to create an efficient tool for development of site-specific WQS in harbors. The integrated model was demonstrated at a harbor-wide scale in San Diego Bay and Pearl Harbor, and accounted for the natural physical, chemical, biological and toxicological characteristics of the harbor to achieve more scientifically based compliance. In both harbors the spatial and temporal distributions of copper species, toxic effects, and Water Effect Ratio predicted by the integrated model are comparable to previous data. The model was further demonstrated in Shelter Island Yacht Basin (SIYB) marina in San Diego Bay. The integrated model agreed with toxicological and chemical approaches by indicating negligible bioavailability as well as no toxicity; but for a single event, even though an increasing gradient in Cu was observed both horizontally and vertically, with concentrations that reached levels well above current regulatory thresholds. These results support the incorporation by USEPA of the seawater-BLM in a full-strength seawater criterion.

SITE TECHNOLOGY CAPSULE: GIS\\KEY ENVIRONMENTAL DATA MANAGEMENT SYSTEM

EPA Science Inventory

GIS/Key™ is a comprehensive environmental database management system that integrates site data and graphics, enabling the user to create geologic cross-sections; boring logs; potentiometric, isopleth, and structure maps; summary tables; and hydrographs. GIS/Key™ is menu-driven an...

Deconstructing thermodynamic parameters of a coupled system from site-specific observables.

PubMed

Chowdhury, Sandipan; Chanda, Baron

2010-11-02

Cooperative interactions mediate information transfer between structural domains of a protein molecule and are major determinants of protein function and modulation. The prevalent theories to understand the thermodynamic origins of cooperativity have been developed to reproduce the complex behavior of a global thermodynamic observable such as ligand binding or enzyme activity. However, in most cases the measurement of a single global observable cannot uniquely define all the terms that fully describe the energetics of the system. Here we establish a theoretical groundwork for analyzing protein thermodynamics using site-specific information. Our treatment involves extracting a site-specific parameter (defined as χ value) associated with a structural unit. We demonstrate that, under limiting conditions, the χ value is related to the direct interaction terms associated with the structural unit under observation and its intrinsic activation energy. We also introduce a site-specific interaction energy term (χ(diff)) that is a function of the direct interaction energy of that site with every other site in the system. When combined with site-directed mutagenesis and other molecular level perturbations, analyses of χ values of site-specific observables may provide valuable insights into protein thermodynamics and structure.

Impacts of climatic change on forest structure in the Midwestern Great Lakes region: An integrated gap model and geographical information system (GIS) approach

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

Eman, J.L.; Randolph, J.C.; Fan, Weihong

The transient effects of an 80-year climate change scenario on 56 tree species are evaluated within the regional forested landscape matrix of Wisconsin, Michigan, Illinois, Indiana, and Ohio. The JABOWA-II forest growth model was integrated with a GIS housing several federally-produced databases which were vital for input file development. A 4km{sup 2} resolution enabled high parameter specificity for each of the 54,345 representative 100m{sup 2} plots. Ten iterations of simulated growth response at each site followed model calibration. Changes in relative regional dominance and shifts in population centroids over two periods (1981-2020 and 2020-2060) indicated that most responses were species-specific.more » Notable among these were the substantial dominance increases of Populus tremuloides and Quercus alba from 1981-2020. Shared response patterns included conifers` centroid shifts to the northwest and overall dominance reductions.« less

Integrating GIS, Archeology, and the Internet.

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

Sera White; Brenda Ringe Pace; Randy Lee

2004-08-01

At the Idaho National Engineering and Environmental Laboratory's (INEEL) Cultural Resource Management Office, a newly developed Data Management Tool (DMT) is improving management and long-term stewardship of cultural resources. The fully integrated system links an archaeological database, a historical database, and a research database to spatial data through a customized user interface using ArcIMS and Active Server Pages. Components of the new DMT are tailored specifically to the INEEL and include automated data entry forms for historic and prehistoric archaeological sites, specialized queries and reports that address both yearly and project-specific documentation requirements, and unique field recording forms. The predictivemore » modeling component increases the DMT’s value for land use planning and long-term stewardship. The DMT enhances the efficiency of archive searches, improving customer service, oversight, and management of the large INEEL cultural resource inventory. In the future, the DMT will facilitate data sharing with regulatory agencies, tribal organizations, and the general public.« less

Multiloop Integral System Test (MIST): MIST Facility Functional Specification

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

Habib, T F; Koksal, C G; Moskal, T E

1991-04-01

The Multiloop Integral System Test (MIST) is part of a multiphase program started in 1983 to address small-break loss-of-coolant accidents (SBLOCAs) specific to Babcock and Wilcox designed plants. MIST is sponsored by the US Nuclear Regulatory Commission, the Babcock Wilcox Owners Group, the Electric Power Research Institute, and Babcock and Wilcox. The unique features of the Babcock and Wilcox design, specifically the hot leg U-bends and steam generators, prevented the use of existing integral system data or existing integral facilities to address the thermal-hydraulic SBLOCA questions. MIST was specifically designed and constructed for this program, and an existing facility --more » the Once Through Integral System (OTIS) -- was also used. Data from MIST and OTIS are used to benchmark the adequacy of system codes, such as RELAP5 and TRAC, for predicting abnormal plant transients. The MIST Functional Specification documents as-built design features, dimensions, instrumentation, and test approach. It also presents the scaling basis for the facility and serves to define the scope of work for the facility design and construction. 13 refs., 112 figs., 38 tabs.« less

Cloning-free CRISPR

PubMed Central

Arbab, Mandana; Srinivasan, Sharanya; Hashimoto, Tatsunori; Geijsen, Niels; Sherwood, Richard I.

2015-01-01

Summary We present self-cloning CRISPR/Cas9 (scCRISPR), a technology that allows for CRISPR/Cas9-mediated genomic mutation and site-specific knockin transgene creation within several hours by circumventing the need to clone a site-specific single-guide RNA (sgRNA) or knockin homology construct for each target locus. We introduce a self-cleaving palindromic sgRNA plasmid and a short double-stranded DNA sequence encoding the desired locus-specific sgRNA into target cells, allowing them to produce a locus-specific sgRNA plasmid through homologous recombination. scCRISPR enables efficient generation of gene knockouts (∼88% mutation rate) at approximately one-sixth the cost of plasmid-based sgRNA construction with only 2 hr of preparation for each targeted site. Additionally, we demonstrate efficient site-specific knockin of GFP transgenes without any plasmid cloning or genome-integrated selection cassette in mouse and human embryonic stem cells (2%–4% knockin rate) through PCR-based addition of short homology arms. scCRISPR substantially lowers the bar on mouse and human transgenesis. PMID:26527385

78 FR 26029 - Toxicological Review of Methanol (Non-Cancer): In Support of Summary Information on the...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-05-03

... of Methanol (Non-Cancer): In Support of Summary Information on the Integrated Risk Information System... Integrated Risk Information System (IRIS)'' (EPA/635/R-11/001Ba), which will be posted on NCEA's Web site... Review of Methanol (Non-Cancer): In Support of Summary Information on the Integrated Risk Information...

Fully integrated automated security surveillance system: managing a changing world through managed technology and product applications

NASA Astrophysics Data System (ADS)

Francisco, Glen; Brown, Todd

2012-06-01

Integrated security systems are essential to pre-empting criminal assaults. Nearly 500,000 sites have been identified (source: US DHS) as critical infrastructure sites that would suffer severe damage if a security breach should occur. One major breach in any of 123 U.S. facilities, identified as "most critical", threatens more than 1,000,000 people. The vulnerabilities of critical infrastructure are expected to continue and even heighten over the coming years.

Develop and test fuel cell powered on-site integrated total energy system

NASA Technical Reports Server (NTRS)

Kaufman, A.; Feigenbaum, H.; Wang, C. L.; Werth, J.; Whelan, J. A.

1983-01-01

Test results are presented for a 24 cell, two sq ft (4kW) stack. This stack is a precursor to a 25kW stack that is a key milestone. Results are discussed in terms of cell performance, electrolyte management, thermal management, and reactant gas manifolding. The results obtained in preliminary testing of a 50kW methanol processing subsystem are discussed. Subcontracting activities involving application analysis for fuel cell on site integrated energy systems are updated.

Lentivector Integration Sites in Ependymal Cells From a Model of Metachromatic Leukodystrophy: Non-B DNA as a New Factor Influencing Integration

PubMed Central

McAllister, Robert G; Liu, Jiahui; Woods, Matthew W; Tom, Sean K; Rupar, C Anthony; Barr, Stephen D

2014-01-01

The blood–brain barrier controls the passage of molecules from the blood into the central nervous system (CNS) and is a major challenge for treatment of neurological diseases. Metachromatic leukodystrophy is a neurodegenerative lysosomal storage disease caused by loss of arylsulfatase A (ARSA) activity. Gene therapy via intraventricular injection of a lentiviral vector is a potential approach to rapidly and permanently deliver therapeutic levels of ARSA to the CNS. We present the distribution of integration sites of a lentiviral vector encoding human ARSA (LV-ARSA) in murine brain choroid plexus and ependymal cells, administered via a single intracranial injection into the CNS. LV-ARSA did not exhibit a strong preference for integration in or near actively transcribed genes, but exhibited a strong preference for integration in or near satellite DNA. We identified several genomic hotspots for LV-ARSA integration and identified a consensus target site sequence characterized by two G-quadruplex-forming motifs flanking the integration site. In addition, our analysis identified several other non-B DNA motifs as new factors that potentially influence lentivirus integration, including human immunodeficiency virus type-1 in human cells. Together, our data demonstrate a clinically favorable integration site profile in the murine brain and identify non-B DNA as a potential new host factor that influences lentiviral integration in murine and human cells. PMID:25158091

System requirement specification for the I-15 integrated corridor management system (ICMS) in San Diego, California.

DOT National Transportation Integrated Search

2008-03-31

This document presents a System Requirement Specification for an Integrated Corridor Management System (ICMS) in the I-15 Corridor in San Diego, California. The ICMS will consist of two major subsystems: the existing Intermodal Transportation Managem...

Conceptions of Connectivity: How Swiss Teachers, Trainers and Apprentices Perceive Vocational Learning and Teaching across Different Learning Sites

ERIC Educational Resources Information Center

Sappa, Viviana; Aprea, Carmela

2014-01-01

The integration of learning across different learning sites is an important and challenging task for each vocational education and training system. Although various integrative teaching and learning models have been developed and many reforms have been undertaken, their implementation is complicated by several factors. In particular, individual…

14. Site D57C, Improved NIKEHercules System, Grading Plan, U.S. Army ...

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

14. Site D-57-C, Improved NIKE-Hercules System, Grading Plan, U.S. Army Corps of Engineers, 18 January 1962. - Newport NIKE Missile Battery D-57/58, Integrated Fire Control Area, Newport Road, Carleton, Monroe County, MI

RED: A Java-MySQL Software for Identifying and Visualizing RNA Editing Sites Using Rule-Based and Statistical Filters.

PubMed

Sun, Yongmei; Li, Xing; Wu, Di; Pan, Qi; Ji, Yuefeng; Ren, Hong; Ding, Keyue

2016-01-01

RNA editing is one of the post- or co-transcriptional processes that can lead to amino acid substitutions in protein sequences, alternative pre-mRNA splicing, and changes in gene expression levels. Although several methods have been suggested to identify RNA editing sites, there remains challenges to be addressed in distinguishing true RNA editing sites from its counterparts on genome and technical artifacts. In addition, there lacks a software framework to identify and visualize potential RNA editing sites. Here, we presented a software - 'RED' (RNA Editing sites Detector) - for the identification of RNA editing sites by integrating multiple rule-based and statistical filters. The potential RNA editing sites can be visualized at the genome and the site levels by graphical user interface (GUI). To improve performance, we used MySQL database management system (DBMS) for high-throughput data storage and query. We demonstrated the validity and utility of RED by identifying the presence and absence of C→U RNA-editing sites experimentally validated, in comparison with REDItools, a command line tool to perform high-throughput investigation of RNA editing. In an analysis of a sample data-set with 28 experimentally validated C→U RNA editing sites, RED had sensitivity and specificity of 0.64 and 0.5. In comparison, REDItools had a better sensitivity (0.75) but similar specificity (0.5). RED is an easy-to-use, platform-independent Java-based software, and can be applied to RNA-seq data without or with DNA sequencing data. The package is freely available under the GPLv3 license at http://github.com/REDetector/RED or https://sourceforge.net/projects/redetector.

RED: A Java-MySQL Software for Identifying and Visualizing RNA Editing Sites Using Rule-Based and Statistical Filters

PubMed Central

Sun, Yongmei; Li, Xing; Wu, Di; Pan, Qi; Ji, Yuefeng; Ren, Hong; Ding, Keyue

2016-01-01

RNA editing is one of the post- or co-transcriptional processes that can lead to amino acid substitutions in protein sequences, alternative pre-mRNA splicing, and changes in gene expression levels. Although several methods have been suggested to identify RNA editing sites, there remains challenges to be addressed in distinguishing true RNA editing sites from its counterparts on genome and technical artifacts. In addition, there lacks a software framework to identify and visualize potential RNA editing sites. Here, we presented a software − ‘RED’ (RNA Editing sites Detector) − for the identification of RNA editing sites by integrating multiple rule-based and statistical filters. The potential RNA editing sites can be visualized at the genome and the site levels by graphical user interface (GUI). To improve performance, we used MySQL database management system (DBMS) for high-throughput data storage and query. We demonstrated the validity and utility of RED by identifying the presence and absence of C→U RNA-editing sites experimentally validated, in comparison with REDItools, a command line tool to perform high-throughput investigation of RNA editing. In an analysis of a sample data-set with 28 experimentally validated C→U RNA editing sites, RED had sensitivity and specificity of 0.64 and 0.5. In comparison, REDItools had a better sensitivity (0.75) but similar specificity (0.5). RED is an easy-to-use, platform-independent Java-based software, and can be applied to RNA-seq data without or with DNA sequencing data. The package is freely available under the GPLv3 license at http://github.com/REDetector/RED or https://sourceforge.net/projects/redetector. PMID:26930599

Biological responses to contaminants in darters (Etheostoma spp.) collected from rural and urban regions of the Grand River, ON, Canada.

PubMed

Diamond, Sam R; Sultana, Tamanna; Servos, Mark R; Metcalfe, Chris D

2016-09-01

Urban and agricultural activities may introduce chemical stressors, including contaminants of emerging concern (CECs) and current use pesticides (CUPs) into riverine systems. The objective of this study was to determine if fish collected from various sites in the Grand River, ON, Canada show biomarkers of exposure to these classes of contaminants, and if the biomarker patterns vary in fish collected from urbanized and agricultural sites. Female rainbow darters (Etheostoma caeruleum) and female fantail darters (Etheostoma flabellare) were collected from the Grand River in June, 2014 for biomarker analysis from two urbanized sites and three agricultural sites. Over the same period of time, Polar Organic Chemical Integrative Samplers (POCIS) were deployed for 2weeks at each site to monitor for the presence of CUPs and CECs. Data on the liver somatic index for darters indicate site-specific differences in this condition factor (p<0.05). Significant differences in the levels of thiobarbituric acid reactive substances (TBARS) in gill tissue (p<0.05) of darters collected from the various sites indicate site-specific differences in oxidative stress. The activities of ethoxyresorufin-O-deethylase (EROD) in the liver tissue of rainbow darters were significantly different between sites (p<0.05), indicating differences in exposure to chemicals that induce or inhibit CYP450 1A metabolic activity. Finally, acetylcholinesterase (AChE) activity in brain tissue was significantly different between rainbow darters collected from rural and urban sites (p<0.05). These data showing different impacts from chemical inputs related to land uses in the watershed may be useful in developing mitigation strategies to reduce impacts on fish and other aquatic organisms in receiving environments. Copyright © 2016. Published by Elsevier Inc.

Engineered catalytic biofilms: Site-specific enzyme immobilization onto E. coli curli nanofibers.

PubMed

Botyanszki, Zsofia; Tay, Pei Kun R; Nguyen, Peter Q; Nussbaumer, Martin G; Joshi, Neel S

2015-10-01

Biocatalytic transformations generally rely on purified enzymes or whole cells to perform complex transformations that are used on industrial scale for chemical, drug, and biofuel synthesis, pesticide decontamination, and water purification. However, both of these systems have inherent disadvantages related to the costs associated with enzyme purification, the long-term stability of immobilized enzymes, catalyst recovery, and compatibility with harsh reaction conditions. We developed a novel strategy for producing rationally designed biocatalytic surfaces based on Biofilm Integrated Nanofiber Display (BIND), which exploits the curli system of E. coli to create a functional nanofiber network capable of covalent immobilization of enzymes. This approach is attractive because it is scalable, represents a modular strategy for site-specific enzyme immobilization, and has the potential to stabilize enzymes under denaturing environmental conditions. We site-specifically immobilized a recombinant α-amylase, fused to the SpyCatcher attachment domain, onto E. coli curli fibers displaying complementary SpyTag capture domains. We characterized the effectiveness of this immobilization technique on the biofilms and tested the stability of immobilized α-amylase in unfavorable conditions. This enzyme-modified biofilm maintained its activity when exposed to a wide range of pH and organic solvent conditions. In contrast to other biofilm-based catalysts, which rely on high cellular metabolism, the modified curli-based biofilm remained active even after cell death due to organic solvent exposure. This work lays the foundation for a new and versatile method of using the extracellular polymeric matrix of E. coli for creating novel biocatalytic surfaces. © 2015 Wiley Periodicals, Inc.

Stakeholder engagement: a key component of integrating genomic information into electronic health records

PubMed Central

Hartzler, Andrea; McCarty, Catherine A.; Rasmussen, Luke V.; Williams, Marc S.; Brilliant, Murray; Bowton, Erica A.; Clayton, Ellen Wright; Faucett, William A.; Ferryman, Kadija; Field, Julie R.; Fullerton, Stephanie M.; Horowitz, Carol R.; Koenig, Barbara A.; McCormick, Jennifer B.; Ralston, James D.; Sanderson, Saskia C.; Smith, Maureen E.; Trinidad, Susan Brown

2014-01-01

Integrating genomic information into clinical care and the electronic health record can facilitate personalized medicine through genetically guided clinical decision support. Stakeholder involvement is critical to the success of these implementation efforts. Prior work on implementation of clinical information systems provides broad guidance to inform effective engagement strategies. We add to this evidence-based recommendations that are specific to issues at the intersection of genomics and the electronic health record. We describe stakeholder engagement strategies employed by the Electronic Medical Records and Genomics Network, a national consortium of US research institutions funded by the National Human Genome Research Institute to develop, disseminate, and apply approaches that combine genomic and electronic health record data. Through select examples drawn from sites of the Electronic Medical Records and Genomics Network, we illustrate a continuum of engagement strategies to inform genomic integration into commercial and homegrown electronic health records across a range of health-care settings. We frame engagement as activities to consult, involve, and partner with key stakeholder groups throughout specific phases of health information technology implementation. Our aim is to provide insights into engagement strategies to guide genomic integration based on our unique network experiences and lessons learned within the broader context of implementation research in biomedical informatics. On the basis of our collective experience, we describe key stakeholder practices, challenges, and considerations for successful genomic integration to support personalized medicine. PMID:24030437

An Integrated Spin-Labeling/Computational-Modeling Approach for Mapping Global Structures of Nucleic Acids.

PubMed

Tangprasertchai, Narin S; Zhang, Xiaojun; Ding, Yuan; Tham, Kenneth; Rohs, Remo; Haworth, Ian S; Qin, Peter Z

2015-01-01

The technique of site-directed spin labeling (SDSL) provides unique information on biomolecules by monitoring the behavior of a stable radical tag (i.e., spin label) using electron paramagnetic resonance (EPR) spectroscopy. In this chapter, we describe an approach in which SDSL is integrated with computational modeling to map conformations of nucleic acids. This approach builds upon a SDSL tool kit previously developed and validated, which includes three components: (i) a nucleotide-independent nitroxide probe, designated as R5, which can be efficiently attached at defined sites within arbitrary nucleic acid sequences; (ii) inter-R5 distances in the nanometer range, measured via pulsed EPR; and (iii) an efficient program, called NASNOX, that computes inter-R5 distances on given nucleic acid structures. Following a general framework of data mining, our approach uses multiple sets of measured inter-R5 distances to retrieve "correct" all-atom models from a large ensemble of models. The pool of models can be generated independently without relying on the inter-R5 distances, thus allowing a large degree of flexibility in integrating the SDSL-measured distances with a modeling approach best suited for the specific system under investigation. As such, the integrative experimental/computational approach described here represents a hybrid method for determining all-atom models based on experimentally-derived distance measurements. © 2015 Elsevier Inc. All rights reserved.

An Integrated Spin-Labeling/Computational-Modeling Approach for Mapping Global Structures of Nucleic Acids

PubMed Central

Tangprasertchai, Narin S.; Zhang, Xiaojun; Ding, Yuan; Tham, Kenneth; Rohs, Remo; Haworth, Ian S.; Qin, Peter Z.

2015-01-01

The technique of site-directed spin labeling (SDSL) provides unique information on biomolecules by monitoring the behavior of a stable radical tag (i.e., spin label) using electron paramagnetic resonance (EPR) spectroscopy. In this chapter, we describe an approach in which SDSL is integrated with computational modeling to map conformations of nucleic acids. This approach builds upon a SDSL tool kit previously developed and validated, which includes three components: (i) a nucleotide-independent nitroxide probe, designated as R5, which can be efficiently attached at defined sites within arbitrary nucleic acid sequences; (ii) inter-R5 distances in the nanometer range, measured via pulsed EPR; and (iii) an efficient program, called NASNOX, that computes inter-R5 distances on given nucleic acid structures. Following a general framework of data mining, our approach uses multiple sets of measured inter-R5 distances to retrieve “correct” all-atom models from a large ensemble of models. The pool of models can be generated independently without relying on the inter-R5 distances, thus allowing a large degree of flexibility in integrating the SDSL-measured distances with a modeling approach best suited for the specific system under investigation. As such, the integrative experimental/computational approach described here represents a hybrid method for determining all-atom models based on experimentally-derived distance measurements. PMID:26477260

System integration of marketable subsystems

NASA Technical Reports Server (NTRS)

1978-01-01

These monthly reports, covering the period February 1978 through June 1978, describe the progress made in the major areas of the program. The areas covered are: systems integration of marketable subsystems; development, design, and building of site data acquisition subsystems; development and operation of the central data processing system; operation of the MSFC Solar Test Facility; and systems analysis.

Enhanced integration of large DNA into E. coli chromosome by CRISPR/Cas9.

PubMed

Chung, Mu-En; Yeh, I-Hsin; Sung, Li-Yu; Wu, Meng-Ying; Chao, Yun-Peng; Ng, I-Son; Hu, Yu-Chen

2017-01-01

Metabolic engineering often necessitates chromosomal integration of multiple genes but integration of large genes into Escherichia coli remains difficult. CRISPR/Cas9 is an RNA-guided system which enables site-specific induction of double strand break (DSB) and programmable genome editing. Here, we hypothesized that CRISPR/Cas9-triggered DSB could enhance homologous recombination and augment integration of large DNA into E. coli chromosome. We demonstrated that CRISPR/Cas9 system was able to trigger DSB in >98% of cells, leading to subsequent cell death, and identified that mutagenic SOS response played roles in the cell survival. By optimizing experimental conditions and combining the λ-Red proteins and linear dsDNA, CRISPR/Cas9-induced DSB enabled homologous recombination of the donor DNA and replacement of lacZ gene in the MG1655 strain at efficiencies up to 99%, and allowed high fidelity, scarless integration of 2.4, 3.9, 5.4, and 7.0 kb DNA at efficiencies approaching 91%, 92%, 71%, and 61%, respectively. The CRISPR/Cas9-assisted gene integration also functioned in different E. coli strains including BL21 (DE3) and W albeit at different efficiencies. Taken together, our methodology facilitated precise integration of dsDNA as large as 7 kb into E. coli with efficiencies exceeding 60%, thus significantly ameliorating the editing efficiency and overcoming the size limit of integration using the commonly adopted recombineering approach. Biotechnol. Bioeng. 2017;114: 172-183. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

SURGNET: An Integrated Surgical Data Transmission System for Telesurgery.

PubMed

Natarajan, Sriram; Ganz, Aura

2009-01-01

Remote surgery information requires quick and reliable transmission between the surgeon and the patient site. However, the networks that interconnect the surgeon and patient sites are usually time varying and lossy which can cause packet loss and delay jitter. In this paper we propose SURGNET, a telesurgery system for which we developed the architecture, algorithms and implemented it on a testbed. The algorithms include adaptive packet prediction and buffer time adjustment techniques which reduce the negative effects caused by the lossy and time varying networks. To evaluate the proposed SURGNET system, at the therapist site, we implemented a therapist panel which controls the force feedback device movements and provides image analysis functionality. At the patient site we controlled a virtual reality applet built in Matlab. The varying network conditions were emulated using NISTNet emulator. Our results show that even for severe packet loss and variable delay jitter, the proposed integrated synchronization techniques significantly improve SURGNET performance.

Characterization of Tactical Departure Scheduling in the National Airspace System

NASA Technical Reports Server (NTRS)

Capps, Alan; Engelland, Shawn A.

2011-01-01

This paper discusses and analyzes current day utilization and performance of the tactical departure scheduling process in the National Airspace System (NAS) to understand the benefits in improving this process. The analysis used operational air traffic data from over 1,082,000 flights during the month of January, 2011. Specific metrics included the frequency of tactical departure scheduling, site specific variances in the technology's utilization, departure time prediction compliance used in the tactical scheduling process and the performance with which the current system can predict the airborne slot that aircraft are being scheduled into from the airport surface. Operational data analysis described in this paper indicates significant room for improvement exists in the current system primarily in the area of reduced departure time prediction uncertainty. Results indicate that a significant number of tactically scheduled aircraft did not meet their scheduled departure slot due to departure time uncertainty. In addition to missed slots, the operational data analysis identified increased controller workload associated with tactical departures which were subject to traffic management manual re-scheduling or controller swaps. An analysis of achievable levels of departure time prediction accuracy as obtained by a new integrated surface and tactical scheduling tool is provided to assess the benefit it may provide as a solution to the identified shortfalls. A list of NAS facilities which are likely to receive the greatest benefit from the integrated surface and tactical scheduling technology are provided.

Integrated platform for optimized solar PV system design and engineering plan set generation

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

Adeyemo, Samuel

2015-12-30

The Aurora team has developed software that allows users to quickly generate a three-dimensional model for a building, with a corresponding irradiance map, from any two-dimensional image with associated geo-coordinates. The purpose of this project is to build upon that technology by developing and distributing to solar installers a software platform that automatically retrieves engineering, financial and geographic data for a specific site, and quickly generates an optimal customer proposal and corresponding engineering plans for that site. At the end of the project, Aurora’s optimization platform would have been used to make at least one thousand proposals from at leastmore » ten unique solar installation companies, two of whom would sign economically viable contracts to use the software. Furthermore, Aurora’s algorithms would be tested to show that in at least seventy percent of cases, Aurora automatically generated a design equivalent to or better than what a human could have done manually. A ‘better’ design is one that generates more energy for the same cost, or that generates a higher return on investment, while complying with all site-specific aesthetic, electrical and spatial requirements.« less

Site-Specific Recombination at XerC/D Sites Mediates the Formation and Resolution of Plasmid Co-integrates Carrying a blaOXA-58- and TnaphA6-Resistance Module in Acinetobacter baumannii

PubMed Central

Cameranesi, María M.; Morán-Barrio, Jorgelina; Limansky, Adriana S.; Repizo, Guillermo D.; Viale, Alejandro M.

2018-01-01

Members of the genus Acinetobacter possess distinct plasmid types which provide effective platforms for the acquisition, evolution, and dissemination of antimicrobial resistance structures. Many plasmid-borne resistance structures are bordered by short DNA sequences providing potential recognition sites for the host XerC and XerD site-specific tyrosine recombinases (XerC/D-like sites). However, whether these sites are active in recombination and how they assist the mobilization of associated resistance structures is still poorly understood. Here we characterized the plasmids carried by Acinetobacter baumannii Ab242, a multidrug-resistant clinical strain belonging to the ST104 (Oxford scheme) which produces an OXA-58 carbapenem-hydrolyzing class-D β-lactamase (CHDL). Plasmid sequencing and characterization of replication, stability, and adaptive modules revealed the presence in Ab242 of three novel plasmids lacking self-transferability functions which were designated pAb242_9, pAb242_12, and pAb242_25, respectively. Among them, only pAb242_25 was found to carry an adaptive module encompassing an ISAba825-blaOXA-58 arrangement accompanied by a TnaphA6 transposon, the whole structure conferring simultaneous resistance to carbapenems and aminoglycosides. Ab242 plasmids harbor several XerC/D-like sites, with most sites found in pAb242_25 located in the vicinity or within the adaptive module described above. Electrotransformation of susceptible A. nosocomialis cells with Ab242 plasmids followed by imipenem selection indicated that the transforming plasmid form was a co-integrate resulting from the fusion of pAb242_25 and pAb242_12. Further characterization by cloning and sequencing studies indicated that a XerC/D site in pAb242_25 and another in pAb242_12 provided the active sister pair for the inter-molecular site-specific recombination reaction mediating the fusion of these two plasmids. Moreover, the resulting co-integrate was found also to undergo intra-molecular resolution at the new pair of XerC/D sites generated during fusion thus regenerating the original pAb242_25 and pAb242_12 plasmids. These observations provide the first evidence indicating that XerC/D-like sites in A. baumannii plasmids can provide active pairs for site-specific recombination mediating inter-molecular fusions and intra-molecular resolutions. The overall results shed light on the evolutionary dynamics of A. baumannii plasmids and the underlying mechanisms of dissemination of genetic structures responsible for carbapenem and other antibiotics resistance among the Acinetobacter clinical population. PMID:29434581

An integrated ecosystem approach for sustainable prevention and control of dengue in Central Havana.

PubMed

Bonet, Mariano; Spiegel, Jerry M; Ibarra, Ana Maria; Kouri, Gustavo; Pintre, Alfredo; Yassi, Annalee

2007-01-01

The authors developed and evaluated a comprehensive participatory ecosystem health approach for preventing the transmission of dengue, the most prevalent vector-borne disease in Cuba and the Latin America-Caribbean region. The integrated surveillance system central to this initiative encompassed three main subsystems (environmental; entomological; clinical-epidemiologic), relying on extensive community involvement. The study was conducted in Central Havana, Cuba. Indicators from each subsystem were selected and mapped using a GIS procedure providing instant visualization by city block in the municipality. To elucidate the factors affecting control and prevention efforts, perceived needs and risks, as well as knowledge, attitudes, and behaviors related to dengue, were assessed. Specific factors associated with the presence of mosquito breeding sites and risks of dengue were examined in a case-control study.

SPEER-SERVER: a web server for prediction of protein specificity determining sites

PubMed Central

Chakraborty, Abhijit; Mandloi, Sapan; Lanczycki, Christopher J.; Panchenko, Anna R.; Chakrabarti, Saikat

2012-01-01

Sites that show specific conservation patterns within subsets of proteins in a protein family are likely to be involved in the development of functional specificity. These sites, generally termed specificity determining sites (SDS), might play a crucial role in binding to a specific substrate or proteins. Identification of SDS through experimental techniques is a slow, difficult and tedious job. Hence, it is very important to develop efficient computational methods that can more expediently identify SDS. Herein, we present Specificity prediction using amino acids’ Properties, Entropy and Evolution Rate (SPEER)-SERVER, a web server that predicts SDS by analyzing quantitative measures of the conservation patterns of protein sites based on their physico-chemical properties and the heterogeneity of evolutionary changes between and within the protein subfamilies. This web server provides an improved representation of results, adds useful input and output options and integrates a wide range of analysis and data visualization tools when compared with the original standalone version of the SPEER algorithm. Extensive benchmarking finds that SPEER-SERVER exhibits sensitivity and precision performance that, on average, meets or exceeds that of other currently available methods. SPEER-SERVER is available at http://www.hpppi.iicb.res.in/ss/. PMID:22689646

SPEER-SERVER: a web server for prediction of protein specificity determining sites.

PubMed

Chakraborty, Abhijit; Mandloi, Sapan; Lanczycki, Christopher J; Panchenko, Anna R; Chakrabarti, Saikat

2012-07-01

Sites that show specific conservation patterns within subsets of proteins in a protein family are likely to be involved in the development of functional specificity. These sites, generally termed specificity determining sites (SDS), might play a crucial role in binding to a specific substrate or proteins. Identification of SDS through experimental techniques is a slow, difficult and tedious job. Hence, it is very important to develop efficient computational methods that can more expediently identify SDS. Herein, we present Specificity prediction using amino acids' Properties, Entropy and Evolution Rate (SPEER)-SERVER, a web server that predicts SDS by analyzing quantitative measures of the conservation patterns of protein sites based on their physico-chemical properties and the heterogeneity of evolutionary changes between and within the protein subfamilies. This web server provides an improved representation of results, adds useful input and output options and integrates a wide range of analysis and data visualization tools when compared with the original standalone version of the SPEER algorithm. Extensive benchmarking finds that SPEER-SERVER exhibits sensitivity and precision performance that, on average, meets or exceeds that of other currently available methods. SPEER-SERVER is available at http://www.hpppi.iicb.res.in/ss/.

Method of making gold thiolate and photochemically functionalized microcantilevers

DOEpatents

Boiadjiev, Vassil I [Knoxville, TN; Brown, Gilbert M [Knoxville, TN; Pinnaduwage, Lal A [Knoxville, TN; Thundat, Thomas G [Knoxville, TN; Bonnesen, Peter V [Knoxville, TN; Goretzki, Gudrun [Nottingham, GB

2009-08-25

Highly sensitive sensor platforms for the detection of specific reagents, such as chromate, gasoline and biological species, using microcantilevers and other microelectromechanical systems (MEMS) whose surfaces have been modified with photochemically attached organic monolayers, such as self-assembled monolayers (SAM), or gold-thiol surface linkage are taught. The microcantilever sensors use photochemical hydrosilylation to modify silicon surfaces and gold-thiol chemistry to modify metallic surfaces thereby enabling individual microcantilevers in multicantilever array chips to be modified separately. Terminal vinyl substituted hydrocarbons with a variety of molecular recognition sites can be attached to the surface of silicon via the photochemical hydrosilylation process. By focusing the activating UV light sequentially on selected silicon or silicon nitride hydrogen terminated surfaces and soaking or spotting selected metallic surfaces with organic thiols, sulfides, or disulfides, the microcantilevers are functionalized. The device and photochemical method are intended to be integrated into systems for detecting specific agents including chromate groundwater contamination, gasoline, and biological species.

Summary of International Waste Management Programs (LLNL Input to SNL L3 MS: System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW)

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

Greenberg, Harris R.; Blink, James A.; Halsey, William G.

2011-08-11

The Used Fuel Disposition Campaign (UFDC) within the Department of Energy’s Office of Nuclear Energy (DOE-NE) Fuel Cycle Technology (FCT) program has been tasked with investigating the disposal of the nation’s spent nuclear fuel (SNF) and high-level nuclear waste (HLW) for a range of potential waste forms and geologic environments. This Lessons Learned task is part of a multi-laboratory effort, with this LLNL report providing input to a Level 3 SNL milestone (System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW). The work package number is: FTLL11UF0328; the work package title is: Technical Bases / Lessons Learned;more » the milestone number is: M41UF032802; and the milestone title is: “LLNL Input to SNL L3 MS: System-Wide Integration and Site Selection Concepts for Future Disposition Options for HLW”. The system-wide integration effort will integrate all aspects of waste management and disposal, integrating the waste generators, interim storage, transportation, and ultimate disposal at a repository site. The review of international experience in these areas is required to support future studies that address all of these components in an integrated manner. Note that this report is a snapshot of nuclear power infrastructure and international waste management programs that is current as of August 2011, with one notable exception. No attempt has been made to discuss the currently evolving world-wide response to the tragic consequences of the earthquake and tsunami that devastated Japan on March 11, 2011, leaving more than 15,000 people dead and more than 8,000 people missing, and severely damaging the Fukushima Daiichi nuclear power complex. Continuing efforts in FY 2012 will update the data, and summarize it in an Excel spreadsheet for easy comparison and assist in the knowledge management of the study cases.« less

Restoration of degraded drylands in northern Chile: The need of local stakeholders' participation to prevent and combat desertification

NASA Astrophysics Data System (ADS)

Jorquera-Jaramillo, Carmen; Yáñez-Acevedo, Marcia; Gutiérrez, Julio R.; Cortés-Bugueño, José Luis; Pastén-Marambio, Víctor; Barraza-Cepeda, Claudia

2015-04-01

Desertification is one of the main factors determining poverty, long-term socio-economic problems, natural resources depletion and disturbances in rural communities living at the Coquimbo Region drylands (North-Central Chile). The Chilean State, along with private initiatives, have invested 473.6 Million dollars (1976 to 2008) to recover degraded areas through afforestation and soil management of 1,373,758 hectares. However, there is no information about the impact of the practices and changes experienced by the local stakeholders. Therefore, there is a need for a comprehensive evaluation considering both socioeconomic and biophysical aspects. To this end, a Protocol on Integrated Assessment (IAPro, PRACTICE project) was applied in two rural communities, involving communal afforested sites and their adjacent degraded drylands: El Sauce (ES, Limarí province) and Las Cañas (LC, Choapa province), Coquimbo Region. Participatory afforestation and soil conservation projects were implemented at both sites by the Chilean National Forestry Service (CONAF) in agreement with each local community (Jiménez y Tapia Agricultural Community at ES and Las Cañas de Choapa Peasant's Community at LC). The protocol involved 7 steps: (1) Stakeholder platform identification and engagement; (2) Baseline assessment and selection of site-specific indicators; (3) Integration and weighting of common and site-specific indicators; (4) Data collection; (5) Integrating and perspectives on a MCDA (Multi-Criteria Decision Analysis); (6) Collective Integrated assessment and knowledge sharing; (7) Dissemination. Interviews involved local and institutional stakeholders related to both sites' implementation, administration and/or local impacts. For the ES site, 5 actions were defined and assessed: No action (control); fences; mechanic and biological practices (soil stabilization, runoff control on slopes); runoff control in micro-basins, gullies and ravines; and footpath for educational and demonstration purposes. The same actions plus 'conservation and management of native forests/shrubs at strict exclosure' were identified at LC site. All of the stakeholders interviewed at both sites recognized the actions, but not all of them had specific knowledge about some of the practices. Afforestation was the most selected and valued practice, revealing deep knowledge about the actions and positive valuation of the site experience, but also a need of more support for maintenance. At ES, the main concerns were drought and derived unemployment; at LC, the main concerns were access to the site, prolonged droughts, local conflicts and lack of community commitment. At both sites, many stakeholders highlighted the need for more financial support, better management and assistance in solving local conflicts. External stakeholders value both sites as replicable models and their positive impacts in increasing afforestation and soil conservation experiences; they also identify a need for continuous financial support and the exploration of other models for site implementation and educational activities. The integrated assessment identified a match between the trends in biophysical indicators' results and the perspectives on those indicators expressed by stakeholders. It remained clear that every intervention to combat desertification requires an active involvement of local stakeholders and the implementation of educational modules to ensure the adoption of appropriate practices. Keywords: Land degradation - restoration actions - participatory approach - economic and environmental aspects - recovery of degraded drylands

Wind Power Forecasting Error Frequency Analyses for Operational Power System Studies: Preprint

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

Florita, A.; Hodge, B. M.; Milligan, M.

2012-08-01

The examination of wind power forecasting errors is crucial for optimal unit commitment and economic dispatch of power systems with significant wind power penetrations. This scheduling process includes both renewable and nonrenewable generators, and the incorporation of wind power forecasts will become increasingly important as wind fleets constitute a larger portion of generation portfolios. This research considers the Western Wind and Solar Integration Study database of wind power forecasts and numerical actualizations. This database comprises more than 30,000 locations spread over the western United States, with a total wind power capacity of 960 GW. Error analyses for individual sites andmore » for specific balancing areas are performed using the database, quantifying the fit to theoretical distributions through goodness-of-fit metrics. Insights into wind-power forecasting error distributions are established for various levels of temporal and spatial resolution, contrasts made among the frequency distribution alternatives, and recommendations put forth for harnessing the results. Empirical data are used to produce more realistic site-level forecasts than previously employed, such that higher resolution operational studies are possible. This research feeds into a larger work of renewable integration through the links wind power forecasting has with various operational issues, such as stochastic unit commitment and flexible reserve level determination.« less

Integrated Imaging Approaches Supporting the Excavation Activities. Multi-Scale Geospatial Documentation in Hierapolis (tk)

NASA Astrophysics Data System (ADS)

Spanò, A.; Chiabrando, F.; Sammartano, G.; Teppati Losè, L.

2018-05-01

The paper focuses on the exploration of the suitability and the discretization of applicability issues about advanced surveying integrated techniques, mainly based on image-based approaches compared and integrated to range-based ones that have been developed with the use of the cutting-edge solutions tested on field. The investigated techniques integrate both technological devices for 3D data acquisition and thus editing and management systems to handle metric models and multi-dimensional data in a geospatial perspective, in order to innovate and speed up the extraction of information during the archaeological excavation activities. These factors, have been experienced in the outstanding site of the Hierapolis of Phrygia ancient city (Turkey), downstream the 2017 surveying missions, in order to produce high-scale metric deliverables in terms of high-detailed Digital Surface Models (DSM), 3D continuous surface models and high-resolution orthoimages products. In particular, the potentialities in the use of UAV platforms for low altitude acquisitions in aerial photogrammetric approach, together with terrestrial panoramic acquisitions (Trimble V10 imaging rover), have been investigated with a comparison toward consolidated Terrestrial Laser Scanning (TLS) measurements. One of the main purposes of the paper is to evaluate the results offered by the technologies used independently and using integrated approaches. A section of the study in fact, is specifically dedicated to experimenting the union of different sensor dense clouds: both dense clouds derived from UAV have been integrated with terrestrial Lidar clouds, to evaluate their fusion. Different test cases have been considered, representing typical situations that can be encountered in archaeological sites.

Toward a Learner-Centered System for Adult Learning

ERIC Educational Resources Information Center

Hermans, Henry; Kalz, Marco; Koper, Rob

2013-01-01

Purpose: The purpose of this paper is to present an e-learning system that integrates the use of concepts of virtual learning environments, personal learning environments, and social network sites. The system is based on a learning model which comprises and integrates three learning contexts for the adult learner: the formal, instructional…

Integrated HTA-FMEA/FMECA methodology for the evaluation of robotic system in urology and general surgery.

PubMed

Frosini, Francesco; Miniati, Roberto; Grillone, Saverio; Dori, Fabrizio; Gentili, Guido Biffi; Belardinelli, Andrea

2016-11-14

The following study proposes and tests an integrated methodology involving Health Technology Assessment (HTA) and Failure Modes, Effects and Criticality Analysis (FMECA) for the assessment of specific aspects related to robotic surgery involving safety, process and technology. The integrated methodology consists of the application of specific techniques coming from the HTA joined to the aid of the most typical models from reliability engineering such as FMEA/FMECA. The study has also included in-site data collection and interviews to medical personnel. The total number of robotic procedures included in the analysis was 44: 28 for urology and 16 for general surgery. The main outcomes refer to the comparative evaluation between robotic, laparoscopic and open surgery. Risk analysis and mitigation interventions come from FMECA application. The small sample size available for the study represents an important bias, especially for the clinical outcomes reliability. Despite this, the study seems to confirm the better trend for robotics' surgical times with comparison to the open technique as well as confirming the robotics' clinical benefits in urology. More complex situation is observed for general surgery, where robotics' clinical benefits directly measured are the lowest blood transfusion rate.

Annual Site Environmental Report: 2008 (ASER)

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

Sabba, D.

2009-11-09

This report provides information about environmental programs during the calendar year of 2008 at the SLAC National Accelerator Laboratory (SLAC), Menlo Park, California. Activities that span the calendar year, i.e., stormwater monitoring covering the winter season of 2008/2009 (October 2008 through May 2009), are also included. Production of an annual site environmental report (ASER) is a requirement established by the United States Department of Energy (DOE) for all management and operating (M&O) contractors throughout the DOE complex. SLAC is a federally-funded research and development center with Stanford University as the M&O contractor. Under Executive Order (EO) 13423, Strengthening Federal Environmental,more » Energy, and Transportation Management, and DOE Order 450.1A, Environmental Protection Program, SLAC effectively implements and integrates the key elements of an Environmental Management System (EMS) to achieve the site's integrated safety and environmental management system goals. For normal daily activities, SLAC managers and supervisors are responsible for ensuring that policies and procedures are understood and followed so that: (1) Worker safety and health are protected; (2) The environment is protected; and (3) Compliance is ensured. Throughout 2008, SLAC continued to improve its management systems. These systems provided a structured framework for SLAC to implement 'greening of the government' initiatives such as EO 13423 and DOE Orders 450.1A and 430.2B. Overall, management systems at SLAC are effective, supporting compliance with all relevant statutory and regulatory requirements. SLAC continues to demonstrate significant progress in implementing and integrating EMS into day-to-day operations and construction activities at SLAC. The annual management review and ranking of environmental aspects were completed this year by SLAC's EMS Steering Committee, the Environmental Safety Committee (ESC), and twelve objectives and targets were established for 2008. For each objective and target, a work plan, or Environmental Management Program (EMP) was completed and progress reports were routinely provided to SLAC senior management and the DOE SLAC Site Office (SSO). During 2008, there were no reportable releases to the environment from SLAC operations. In addition, many improvements in waste minimization, recycling, stormwater management, groundwater restoration, and SLAC's chemical management system (CMS) were continued during the year. The following are amongst SLAC's environmental accomplishments for 2008: a composting program at SLAC's onsite cafeteria was initiated, greater than 800 cubic feet of legacy radioactive waste were packaged and shipped from SLAC, a chemical redistribution program was developed, SLAC reduced the number of General Services Administration leased vehicles from 221 to 164, recycling of municipal waste was increased by approximately 140 tons during 2008, and site-wide releases of sulfur hexafluoride were reduced by 50 percent. In 2008, no radiological incidents occurred that increased radiation levels or released radioactivity to the environment. In addition to managing its radioactive wastes safely and responsibly, SLAC worked to reduce the amount of waste generated. SLAC has implemented programs and systems to ensure compliance with all radiological requirements related to the environment. Specifically, the Radiation Protection Radiological Waste Management Group developed a training course to certify Radioactive Waste Generators, conducted a training pilot, and developed a list of potential radioactive waste generators to train. Twenty eight generators were trained in 2008. As a best management practice, SLAC also reduced its tritium inventory by at least 95 percent by draining one of its accelerator cooling water systems; with the cooperation of the South Bayside System Authority, the West Bay Sanitary District and the DOE, SLAC discharged the cooling water to the sanitary sewer according to federal regulations and replenished the system with clean water. In 2008, the SLAC Environmental Restoration Program personnel continued work on site characterization and evaluation of remedial alternatives at four sites with volatile organic compounds in groundwater and several areas with polychlorinated biphenyls and low concentrations of lead in soil. SLAC is regulated under a site cleanup requirements order (board order) issued by the California Regional Water Quality Control Board (RWQCB), San Francisco Bay Region in May 2005 for the investigation and remediation of impacted soil and groundwater at SLAC. The board order lists specific tasks and deadlines for completion of groundwater and soil characterization and other remediation activities. All 2008 submittals to the RWQCB were completed and submitted on time.« less

Service system integration, access to services, and housing outcomes in a program for homeless persons with severe mental illness.

PubMed

Rosenheck, R; Morrissey, J; Lam, J; Calloway, M; Johnsen, M; Goldman, H; Randolph, F; Blasinsky, M; Fontana, A; Calsyn, R; Teague, G

1998-11-01

This study evaluated the hypothesis that greater integration and coordination between agencies within service systems is associated with greater accessibility of services and improved client housing outcomes. As part of the Access to Community Care and Effective Services and Supports program, data were obtained on baseline client characteristics, service use, and 3-month and 12-month outcomes from 1832 clients seen at 18 sites during the first year of program operation. Data on interorganizational relationships were obtained from structured interviews with key informants from relevant organizations in each community (n = 32-82 at each site). Complete follow-up data were obtained from 1340 clients (73%). After control for baseline characteristics, service system integration was associated with superior housing outcomes at 12 months, and this relationship was mediated through greater access to housing agencies. Service system integration is related to improved access to housing services and better housing outcomes among homeless people with mental illness.

DICOM index tracker enterprise: advanced system for enterprise-wide quality assurance and patient safety monitoring

NASA Astrophysics Data System (ADS)

Zhang, Min; Pavlicek, William; Panda, Anshuman; Langer, Steve G.; Morin, Richard; Fetterly, Kenneth A.; Paden, Robert; Hanson, James; Wu, Lin-Wei; Wu, Teresa

2015-03-01

DICOM Index Tracker (DIT) is an integrated platform to harvest rich information available from Digital Imaging and Communications in Medicine (DICOM) to improve quality assurance in radiology practices. It is designed to capture and maintain longitudinal patient-specific exam indices of interests for all diagnostic and procedural uses of imaging modalities. Thus, it effectively serves as a quality assurance and patient safety monitoring tool. The foundation of DIT is an intelligent database system which stores the information accepted and parsed via a DICOM receiver and parser. The database system enables the basic dosimetry analysis. The success of DIT implementation at Mayo Clinic Arizona calls for the DIT deployment at the enterprise level which requires significant improvements. First, for geographically distributed multi-site implementation, the first bottleneck is the communication (network) delay; the second is the scalability of the DICOM parser to handle the large volume of exams from different sites. To address this issue, DICOM receiver and parser are separated and decentralized by site. To facilitate the enterprise wide Quality Assurance (QA), a notable challenge is the great diversities of manufacturers, modalities and software versions, as the solution DIT Enterprise provides the standardization tool for device naming, protocol naming, physician naming across sites. Thirdly, advanced analytic engines are implemented online which support the proactive QA in DIT Enterprise.

Groundwork for Universal Canister System Development

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

Price, Laura L.; Gross, Mike; Prouty, Jeralyn L.

2015-09-01

The mission of the United States Department of Energy's Office of Environmental Management is to complete the safe cleanup of the environmental legacy brought about from five decades of nuclear weapons development and go vernment - sponsored nuclear energy re search. S ome of the waste s that that must be managed have be en identified as good candidates for disposal in a deep borehole in crystalline rock (SNL 2014 a). In particular, wastes that can be disposed of in a small package are good candidates for this disposal concept. A canister - based system that can be used formore » handling these wastes during the disposition process (i.e., storage, transfers, transportation, and disposal) could facilitate the eventual disposal of these wastes. This report provides information for a program plan for developing specifications regarding a canister - based system that facilitates small waste form packaging and disposal and that is integrated with the overall efforts of the DOE's Office of Nuclear Energy Used Fuel Dis position Camp aign's Deep Borehole Field Test . Groundwork for Universal Ca nister System Development September 2015 ii W astes to be considered as candidates for the universal canister system include capsules containing cesium and strontium currently stored in pools at the Hanford Site, cesium to be processed using elutable or nonelutable resins at the Hanford Site, and calcine waste from Idaho National Laboratory. The initial emphasis will be on disposal of the cesium and strontium capsules in a deep borehole that has been drilled into crystalline rock. Specifications for a universal canister system are derived from operational, performance, and regulatory requirements for storage, transfers, transportation, and disposal of radioactive waste. Agreements between the Department of Energy and the States of Washington and Idaho, as well as the Deep Borehole Field Test plan provide schedule requirements for development of the universal canister system . Future work includes collaboration with the Hanford Site to move the cesium and strontium capsules into dry storage, collaboration with the Deep Borehole Field Tes t to develop surface handling and emplacement techniques and to develop the waste package design requirements, developing universal canister system design options and concepts of operations, and developing system analysis tools. Areas in which f urther research and development are needed include material properties and structural integrity, in - package sorbents and fillers, waste form tolerance to heat and postweld stress relief, waste package impact limiters, sensors, cesium mobility under downhol e conditions, and the impact of high pressure and high temperature environment on seals design.« less

Integrating biofiltration with SVE: A case study

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

Lesley, M.P.; Rangan, C.R.

1996-12-01

A prototype integrated soil vacuum extraction/biofiltration system has been designed and installed at a gasoline contaminated LUST site in southern Delaware. The prototype system remediates contaminated moisture entrained in the air stream, employs automatic water level controls in the filters, and achieves maximum vapor extraction and VOC destruction efficiency with an optimum power input. In addition, the valving and piping layout allows the direction of air flow through the filters to be reversed at a given time interval, which minimizes biofouling, thereby increasing efficiency by minimizing the need for frequent cleaning. This integrated system achieves constant VOC destruction rates ofmore » 40 to 70% while maintaining optimal VOC removal rates from the subsurface. The modular design allows for easy mobilization, setup and demobilization at state-lead LUST sites throughout Delaware.« less

A novel, broad-range, CTXΦ-derived stable integrative expression vector for functional studies.

PubMed

Das, Bhabatosh; Kumari, Reena; Pant, Archana; Sen Gupta, Sourav; Saxena, Shruti; Mehta, Ojasvi; Nair, Gopinath Balakrish

2014-12-01

CTXΦ, a filamentous vibriophage encoding cholera toxin, uses a unique strategy for its lysogeny. The single-stranded phage genome forms intramolecular base-pairing interactions between two inversely oriented XerC and XerD binding sites (XBS) and generates a functional phage attachment site, attP(+), for integration. The attP(+) structure is recognized by the host-encoded tyrosine recombinases XerC and XerD (XerCD), which enables irreversible integration of CTXΦ into the chromosome dimer resolution site (dif) of Vibrio cholerae. The dif site and the XerCD recombinases are widely conserved in bacteria. We took advantage of these conserved attributes to develop a broad-host-range integrative expression vector that could irreversibly integrate into the host chromosome using XerCD recombinases without altering the function of any known open reading frame (ORF). In this study, we engineered two different arabinose-inducible expression vectors, pBD62 and pBD66, using XBS of CTXΦ. pBD62 replicates conditionally and integrates efficiently into the dif of the bacterial chromosome by site-specific recombination using host-encoded XerCD recombinases. The expression level of the gene of interest could be controlled through the PBAD promoter by modulating the functions of the vector-encoded transcriptional factor AraC. We validated the irreversible integration of pBD62 into a wide range of pathogenic and nonpathogenic bacteria, such as V. cholerae, Vibrio fluvialis, Vibrio parahaemolyticus, Escherichia coli, Salmonella enterica, and Klebsiella pneumoniae. Gene expression from the PBAD promoter of integrated vectors was confirmed in V. cholerae using the well-studied reporter genes mCherry, eGFP, and lacZ. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Water quality monitoring strategies - A review and future perspectives.

PubMed

Behmel, S; Damour, M; Ludwig, R; Rodriguez, M J

2016-11-15

The reliable assessment of water quality through water quality monitoring programs (WQMPs) is crucial in order for decision-makers to understand, interpret and use this information in support of their management activities aiming at protecting the resource. The challenge of water quality monitoring has been widely addressed in the literature since the 1940s. However, there is still no generally accepted, holistic and practical strategy to support all phases of WQMPs. The purpose of this paper is to report on the use cases a watershed manager has to address to plan or optimize a WQMP from the challenge of identifying monitoring objectives; selecting sampling sites and water quality parameters; identifying sampling frequencies; considering logistics and resources to the implementation of actions based on information acquired through the WQMP. An inventory and critique of the information, approaches and tools placed at the disposal of watershed managers was proposed to evaluate how the existing information could be integrated in a holistic, user-friendly and evolvable solution. Given the differences in regulatory requirements, water quality standards, geographical and geological differences, land-use variations, and other site specificities, a one-in-all solution is not possible. However, we advance that an intelligent decision support system (IDSS) based on expert knowledge that integrates existing approaches and past research can guide a watershed manager through the process according to his/her site-specific requirements. It is also necessary to tap into local knowledge and to identify the knowledge needs of all the stakeholders through participative approaches based on geographical information systems and adaptive survey-based questionnaires. We believe that future research should focus on developing such participative approaches and further investigate the benefits of IDSS's that can be updated quickly and make it possible for a watershed manager to obtain a timely, holistic view and support for every aspect of planning and optimizing a WQMP. Copyright © 2016 Elsevier B.V. All rights reserved.

FLP recombinase-mediated site-specific recombination in silkworm, Bombyx mori

USDA-ARS?s Scientific Manuscript database

A comprehensive understanding of gene function and the production of site-specific genetically modified mutants are two major goals of genetic engineering in the post-genomic era. Although site-specific recombination systems have been powerful tools for genome manipulation of many organisms, they h...

High-efficiency transformation of Pichia stipitis based on its URA3 gene and a homologous autonomous replication sequence, ARS2.

PubMed Central

Yang, V W; Marks, J A; Davis, B P; Jeffries, T W

1994-01-01

This paper describes the first high-efficiency transformation system for the xylose-fermenting yeast Pichia stipitis. The system includes integrating and autonomously replicating plasmids based on the gene for orotidine-5'-phosphate decarboxylase (URA3) and an autonomous replicating sequence (ARS) element (ARS2) isolated from P. stipitis CBS 6054. Ura- auxotrophs were obtained by selecting for resistance to 5-fluoroorotic acid and were identified as ura3 mutants by transformation with P. stipitis URA3. P. stipitis URA3 was cloned by its homology to Saccharomyces cerevisiae URA3, with which it is 69% identical in the coding region. P. stipitis ARS elements were cloned functionally through plasmid rescue. These sequences confer autonomous replication when cloned into vectors bearing the P. stipitis URA3 gene. P. stipitis ARS2 has features similar to those of the consensus ARS of S. cerevisiae and other ARS elements. Circular plasmids bearing the P. stipitis URA3 gene with various amounts of flanking sequences produced 600 to 8,600 Ura+ transformants per micrograms of DNA by electroporation. Most transformants obtained with circular vectors arose without integration of vector sequences. One vector yielded 5,200 to 12,500 Ura+ transformants per micrograms of DNA after it was linearized at various restriction enzyme sites within the P. stipitis URA3 insert. Transformants arising from linearized vectors produced stable integrants, and integration events were site specific for the genomic ura3 in 20% of the transformants examined. Plasmids bearing the P. stipitis URA3 gene and ARS2 element produced more than 30,000 transformants per micrograms of plasmid DNA. Autonomously replicating plasmids were stable for at least 50 generations in selection medium and were present at an average of 10 copies per nucleus. Images PMID:7811063

Opportunities to integrate solar technologies into the Chilean lithium mining industry - reducing process related GHG emissions of a strategic storage resource

NASA Astrophysics Data System (ADS)

Telsnig, Thomas; Potz, Christian; Haas, Jannik; Eltrop, Ludger; Palma-Behnke, Rodrigo

2017-06-01

The arid northern regions of Chile are characterized by an intensive mineral mining industry and high solar irradiance levels. Besides Chile's main mining products, copper, molybdenum and iron, the production of lithium carbonate from lithium containing brines has become strategically important due to the rising demand for battery technologies worldwide. Its energy-intensive production may affect the ecological footprint of the product and the country's climate targets. Thus, the use of solar technologies for electricity and heat production might constitute an interesting option for CO2 mitigation. This study aims to quantify the impacts of the lithium carbonate production processes in Chile on climate change, and to identify site-specific integration options of solar energy technologies to reduce GHG life-cycle emissions. The considered solar integration options include a parabolic trough power plant with a molten salt storage, a solar tower power plant with molten salt receiver and molten salt storage, a one-axis tracking photovoltaic energy system for electricity, and two solar thermal power plants with Ruths storage (steam accumulator) for thermal heat production. CSP plants were identified as measures with the highest GHG mitigation potential reducing the CO2 emissions for the entire production chain and the lithium production between 16% and 33%. In a scenario that combines solar technologies for electricity and thermal energy generation, up to 59% of the CO2 emissions at the lithium production sites in Chile can be avoided. A comparison of the GHG abatement costs of the proposed solar integration options indicates that the photovoltaic system, the solar thermal plant with limited storage and the solar tower power plant are the most cost effective options.

Site-specific recombination in the chicken genome using Flipase recombinase-mediated cassette exchange.

PubMed

Lee, Hong Jo; Lee, Hyung Chul; Kim, Young Min; Hwang, Young Sun; Park, Young Hyun; Park, Tae Sub; Han, Jae Yong

2016-02-01

Targeted genome recombination has been applied in diverse research fields and has a wide range of possible applications. In particular, the discovery of specific loci in the genome that support robust and ubiquitous expression of integrated genes and the development of genome-editing technology have facilitated rapid advances in various scientific areas. In this study, we produced transgenic (TG) chickens that can induce recombinase-mediated gene cassette exchange (RMCE), one of the site-specific recombination technologies, and confirmed RMCE in TG chicken-derived cells. As a result, we established TG chicken lines that have, Flipase (Flp) recognition target (FRT) pairs in the chicken genome, mediated by piggyBac transposition. The transgene integration patterns were diverse in each TG chicken line, and the integration diversity resulted in diverse levels of expression of exogenous genes in each tissue of the TG chickens. In addition, the replaced gene cassette was expressed successfully and maintained by RMCE in the FRT predominant loci of TG chicken-derived cells. These results indicate that targeted genome recombination technology with RMCE could be adaptable to TG chicken models and that the technology would be applicable to specific gene regulation by cis-element insertion and customized expression of functional proteins at predicted levels without epigenetic influence. © FASEB.

HTLV-1 Integration into Transcriptionally Active Genomic Regions Is Associated with Proviral Expression and with HAM/TSP

PubMed Central

Meekings, Kiran N.; Leipzig, Jeremy; Bushman, Frederic D.; Taylor, Graham P.; Bangham, Charles R. M.

2008-01-01

Human T-lymphotropic virus type 1 (HTLV-1) causes leukaemia or chronic inflammatory disease in ∼5% of infected hosts. The level of proviral expression of HTLV-1 differs significantly among infected people, even at the same proviral load (proportion of infected mononuclear cells in the circulation). A high level of expression of the HTLV-1 provirus is associated with a high proviral load and a high risk of the inflammatory disease of the central nervous system known as HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). But the factors that control the rate of HTLV-1 proviral expression remain unknown. Here we show that proviral integration sites of HTLV-1 in vivo are not randomly distributed within the human genome but are associated with transcriptionally active regions. Comparison of proviral integration sites between individuals with high and low levels of proviral expression, and between provirus-expressing and provirus non-expressing cells from within an individual, demonstrated that frequent integration into transcription units was associated with an increased rate of proviral expression. An increased frequency of integration sites in transcription units in individuals with high proviral expression was also associated with the inflammatory disease HAM/TSP. By comparing the distribution of integration sites in human lymphocytes infected in short-term cell culture with those from persistent infection in vivo, we infer the action of two selective forces that shape the distribution of integration sites in vivo: positive selection for cells containing proviral integration sites in transcriptionally active regions of the genome, and negative selection against cells with proviral integration sites within transcription units. PMID:18369476

Telemedicine and security. Confidentiality, integrity, and availability: a Canadian perspective.

PubMed

Jennett, P; Watanabe, M; Igras, E; Premkumar, K; Hall, W

1996-01-01

The health care system is undergoing major reform, characterized by organized delivery systems (regionalization, decentralization, devolution, etc); shifts in care delivery sites; changing health provider roles; increasing consumer responsibilities; and accountability. Rapid advances in information technology and telecommunications have led to a new type of information infrastructure which can play a major role in this reform. Compatible health information systems are now being integrated and connected across institutional, regional, and sectorial boundaries. In the near future, these information systems will readily be accessed and shared by health providers, researchers, policy makers, health consumers, and the public. SECURITY is a critical characteristic of any health information system. This paper will address three fields associated with SECURITY: confidentiality, integrity, and availability. These will be defined and examined as they relate to specific aspects of Telemedicine, such as electronic integrated records and clinical databases; electronic transfer of documents; as well as data storage and disposal. The guiding principles, standards, and safeguards being considered and put in place to ensure that telemedicine information intrastructures can protect and benefit all stakeholders' rights and needs in both primary and secondary uses of information will be reviewed. Implemented, proposed, and tested institutional, System, and Network solutions will be discussed; for example, encryption-decryption methods; data transfer standards; individual and terminal access and entry I.D. and password levels; smart card access and PIN number control; data loss prevention strategies; interference alerts; information access keys; algorithm safeguards; and active marketing to users of standards and principles. Issues such as policy, implementation, and ownership will be addressed.

20--500 watt AMTEC auxiliary electric power system

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

Ivanenok, J.F. III; Sievers, R.K.

1996-12-31

Numerous design studies have been completed on Alkali Metal Thermal to Electric Converter (AMTEC) power systems for space applications demonstrating their substantial increase in performance. Recently design studies have been initiated to couple AMTEC power conversion with fossil fueled combustion systems. This paper describes the results of a Phase 1 SBIR effort to design an innovative, efficient, reliable, long life AMTEC Auxiliary Electric Power System (AEPS) for remote site applications (20--500 watts). The concept uses high voltage AMTEC cells, each containing 7 to 9 small electrolyte tubes, integrated with a combustor and recuperator. These multi-tube AMTEC cells are low cost,more » reliable, long life static converters. AMTEC technology is ideal for auxiliary electric power supplies that must operate reliably over a broad range of temperatures, fuel sources, power levels, and operational specifications. The simplicity, efficiency (20% systems) and modularity of this technology allow it to fill applications as varied as light-weight backpacks, remote site power supplies, and military base power. Phase 1 demonstrated the feasibility of a 20% system design, and showed that the development needs to focus on identifying long life AMTEC cell components, determining the AMTEC cell and system reliability, and demonstrating that a 20 watt AMTEC system is 3--5 times more efficient than existing systems for the same application.« less

Integrated system for gathering, processing, and reporting data relating to site contamination

DOEpatents

Long, D.D.; Goldberg, M.S.; Baker, L.A.

1997-11-11

An integrated screening system comprises an intrusive sampling subsystem, a field mobile laboratory subsystem, a computer assisted design/geographical information subsystem, and a telecommunication linkup subsystem, all integrated to provide synergistically improved data relating to the extent of site soil/groundwater contamination. According to the present invention, data samples related to the soil, groundwater or other contamination of the subsurface material are gathered and analyzed to measure contaminants. Based on the location of origin of the samples in three-dimensional space, the analyzed data are transmitted to a location display. The data from analyzing samples and the data from the locating the origin are managed to project the next probable sample location. The next probable sample location is then forwarded for use as a guide in the placement of ensuing sample location, whereby the number of samples needed to accurately characterize the site is minimized. 10 figs.

Integrated system for gathering, processing, and reporting data relating to site contamination

DOEpatents

Long, Delmar D.; Goldberg, Mitchell S.; Baker, Lorie A.

1997-01-01

An integrated screening system comprises an intrusive sampling subsystem, a field mobile laboratory subsystem, a computer assisted design/geographical information subsystem, and a telecommunication linkup subsystem, all integrated to provide synergistically improved data relating to the extent of site soil/groundwater contamination. According to the present invention, data samples related to the soil, groundwater or other contamination of the subsurface material are gathered and analyzed to measure contaminants. Based on the location of origin of the samples in three-dimensional space, the analyzed data are transmitted to a location display. The data from analyzing samples and the data from the locating the origin are managed to project the next probable sample location. The next probable sample location is then forwarded for use as a guide in the placement of ensuing sample location, whereby the number of samples needed to accurately characterize the site is minimized.

Linking terrestrial and marine conservation planning and threats analysis.

PubMed

Tallis, Heather; Ferdaña, Zach; Gray, Elizabeth

2008-02-01

The existence of the Gulf of Mexico dead zone makes it clear that marine ecosystems can be damaged by terrestrial inputs. Marine and terrestrial conservation planning need to be aligned in an explicit fashion to fully represent threats to marine systems. To integrate conservation planning for terrestrial and marine systems, we used a novel threats assessment that included 5 cross-system threats in a site-prioritization exercise for the Pacific Northwest coast ecoregion (U.S.A.). Cross-system threats are actions or features in one ecological realm that have effects on species in another realm. We considered bulkheads and other forms of shoreline hardening threats to terrestrial systems and roads, logging, agriculture, and urban areas threats to marine systems. We used 2 proxies of freshwater influence on marine environments, validated against a mechanistic model and field observations, to propagate land-based threats into marine sites. We evaluated the influence of cross-system threats on conservation priorities by comparing MARXAN outputs for 3 scenarios that identified terrestrial and marine priorities simultaneously: (1) no threats, (2) single-system threats, and (3) single- and cross-system threats. Including cross-system threats changed the threat landscape dramatically. As a result the best plan that included only single-system threats identified 323 sites (161,500 ha) at risk from cross-system threats. Including these threats changed the location of best sites. By comparing the best and sum solutions of the single- and cross-system scenarios, we identified areas ideal for preservation or restoration through integrated management. Our findings lend quantitative support to the call for explicitly integrated decision making and management action in terrestrial and marine ecosystems.

Adeno-Associated Virus Type 2 Wild-Type and Vector-Mediated Genomic Integration Profiles of Human Diploid Fibroblasts Analyzed by Third-Generation PacBio DNA Sequencing

PubMed Central

Hüser, Daniela; Gogol-Döring, Andreas; Chen, Wei

2014-01-01

ABSTRACT Genome-wide analysis of adeno-associated virus (AAV) type 2 integration in HeLa cells has shown that wild-type AAV integrates at numerous genomic sites, including AAVS1 on chromosome 19q13.42. Multiple GAGY/C repeats, resembling consensus AAV Rep-binding sites are preferred, whereas rep-deficient AAV vectors (rAAV) regularly show a random integration profile. This study is the first study to analyze wild-type AAV integration in diploid human fibroblasts. Applying high-throughput third-generation PacBio-based DNA sequencing, integration profiles of wild-type AAV and rAAV are compared side by side. Bioinformatic analysis reveals that both wild-type AAV and rAAV prefer open chromatin regions. Although genomic features of AAV integration largely reproduce previous findings, the pattern of integration hot spots differs from that described in HeLa cells before. DNase-Seq data for human fibroblasts and for HeLa cells reveal variant chromatin accessibility at preferred AAV integration hot spots that correlates with variant hot spot preferences. DNase-Seq patterns of these sites in human tissues, including liver, muscle, heart, brain, skin, and embryonic stem cells further underline variant chromatin accessibility. In summary, AAV integration is dependent on cell-type-specific, variant chromatin accessibility leading to random integration profiles for rAAV, whereas wild-type AAV integration sites cluster near GAGY/C repeats. IMPORTANCE Adeno-associated virus type 2 (AAV) is assumed to establish latency by chromosomal integration of its DNA. This is the first genome-wide analysis of wild-type AAV2 integration in diploid human cells and the first to compare wild-type to recombinant AAV vector integration side by side under identical experimental conditions. Major determinants of wild-type AAV integration represent open chromatin regions with accessible consensus AAV Rep-binding sites. The variant chromatin accessibility of different human tissues or cell types will have impact on vector targeting to be considered during gene therapy. PMID:25031342

IHE cross-enterprise document sharing for imaging: interoperability testing software

PubMed Central

2010-01-01

Background With the deployments of Electronic Health Records (EHR), interoperability testing in healthcare is becoming crucial. EHR enables access to prior diagnostic information in order to assist in health decisions. It is a virtual system that results from the cooperation of several heterogeneous distributed systems. Interoperability between peers is therefore essential. Achieving interoperability requires various types of testing. Implementations need to be tested using software that simulates communication partners, and that provides test data and test plans. Results In this paper we describe a software that is used to test systems that are involved in sharing medical images within the EHR. Our software is used as part of the Integrating the Healthcare Enterprise (IHE) testing process to test the Cross Enterprise Document Sharing for imaging (XDS-I) integration profile. We describe its architecture and functionalities; we also expose the challenges encountered and discuss the elected design solutions. Conclusions EHR is being deployed in several countries. The EHR infrastructure will be continuously evolving to embrace advances in the information technology domain. Our software is built on a web framework to allow for an easy evolution with web technology. The testing software is publicly available; it can be used by system implementers to test their implementations. It can also be used by site integrators to verify and test the interoperability of systems, or by developers to understand specifications ambiguities, or to resolve implementations difficulties. PMID:20858241

IHE cross-enterprise document sharing for imaging: interoperability testing software.

PubMed

Noumeir, Rita; Renaud, Bérubé

2010-09-21

With the deployments of Electronic Health Records (EHR), interoperability testing in healthcare is becoming crucial. EHR enables access to prior diagnostic information in order to assist in health decisions. It is a virtual system that results from the cooperation of several heterogeneous distributed systems. Interoperability between peers is therefore essential. Achieving interoperability requires various types of testing. Implementations need to be tested using software that simulates communication partners, and that provides test data and test plans. In this paper we describe a software that is used to test systems that are involved in sharing medical images within the EHR. Our software is used as part of the Integrating the Healthcare Enterprise (IHE) testing process to test the Cross Enterprise Document Sharing for imaging (XDS-I) integration profile. We describe its architecture and functionalities; we also expose the challenges encountered and discuss the elected design solutions. EHR is being deployed in several countries. The EHR infrastructure will be continuously evolving to embrace advances in the information technology domain. Our software is built on a web framework to allow for an easy evolution with web technology. The testing software is publicly available; it can be used by system implementers to test their implementations. It can also be used by site integrators to verify and test the interoperability of systems, or by developers to understand specifications ambiguities, or to resolve implementations difficulties.

A technical, economic, and environmental assessment of amine-based CO2 capture technology for power plant greenhouse gas control.

PubMed

Rao, Anand B; Rubin, Edward S

2002-10-15

Capture and sequestration of CO2 from fossil fuel power plants is gaining widespread interest as a potential method of controlling greenhouse gas emissions. Performance and cost models of an amine (MEA)-based CO2 absorption system for postcombustion flue gas applications have been developed and integrated with an existing power plant modeling framework that includes multipollutant control technologies for other regulated emissions. The integrated model has been applied to study the feasibility and cost of carbon capture and sequestration at both new and existing coal-burning power plants. The cost of carbon avoidance was shown to depend strongly on assumptions about the reference plant design, details of the CO2 capture system design, interactions with other pollution control systems, and method of CO2 storage. The CO2 avoidance cost for retrofit systems was found to be generally higher than for new plants, mainly because of the higher energy penalty resulting from less efficient heat integration as well as site-specific difficulties typically encountered in retrofit applications. For all cases, a small reduction in CO2 capture cost was afforded by the SO2 emission trading credits generated by amine-based capture systems. Efforts are underway to model a broader suite of carbon capture and sequestration technologies for more comprehensive assessments in the context of multipollutant environmental management.

Site-specific recombination for precise and clean transgene integration in plant genome. In: Touraev, A., Citovsky, V., Tzfira, T., Editors of book. Plant Transformation Technologies.

USDA-ARS?s Scientific Manuscript database

Plant Transformation Technologies is a comprehensive, authoritative book focusing on cutting-edge plant biotechnologies, offering in-depth, forward-looking information on methods for controlled and accurate genetic engineering. In response to ever-increasing pressure for precise and efficient integr...

Enterprise networks. Strategies for integrated delivery systems.

PubMed

Siwicki, B

1997-02-01

More integrated delivery systems are making progress toward building computer networks that link all their care delivery sites so they can efficiently and economically coordinate care. A growing number of these systems are turning to intranets--private computer networks that use Internet-derived protocols and technologies--to move information that's essential to managing scare health care resources.

A new spatial multi-criteria decision support tool for site selection for implementation of managed aquifer recharge.

PubMed

Rahman, M Azizur; Rusteberg, Bernd; Gogu, R C; Lobo Ferreira, J P; Sauter, Martin

2012-05-30

This study reports the development of a new spatial multi-criteria decision analysis (SMCDA) software tool for selecting suitable sites for Managed Aquifer Recharge (MAR) systems. The new SMCDA software tool functions based on the combination of existing multi-criteria evaluation methods with modern decision analysis techniques. More specifically, non-compensatory screening, criteria standardization and weighting, and Analytical Hierarchy Process (AHP) have been combined with Weighted Linear Combination (WLC) and Ordered Weighted Averaging (OWA). This SMCDA tool may be implemented with a wide range of decision maker's preferences. The tool's user-friendly interface helps guide the decision maker through the sequential steps for site selection, those steps namely being constraint mapping, criteria hierarchy, criteria standardization and weighting, and criteria overlay. The tool offers some predetermined default criteria and standard methods to increase the trade-off between ease-of-use and efficiency. Integrated into ArcGIS, the tool has the advantage of using GIS tools for spatial analysis, and herein data may be processed and displayed. The tool is non-site specific, adaptive, and comprehensive, and may be applied to any type of site-selection problem. For demonstrating the robustness of the new tool, a case study was planned and executed at Algarve Region, Portugal. The efficiency of the SMCDA tool in the decision making process for selecting suitable sites for MAR was also demonstrated. Specific aspects of the tool such as built-in default criteria, explicit decision steps, and flexibility in choosing different options were key features, which benefited the study. The new SMCDA tool can be augmented by groundwater flow and transport modeling so as to achieve a more comprehensive approach to the selection process for the best locations of the MAR infiltration basins, as well as the locations of recovery wells and areas of groundwater protection. The new spatial multicriteria analysis tool has already been implemented within the GIS based Gabardine decision support system as an innovative MAR planning tool. Copyright © 2012 Elsevier Ltd. All rights reserved.

An operational centre for managing major chemical industrial accidents.

PubMed

Kiranoudis, C T; Kourniotis, S P; Christolis, M; Markatos, N C; Zografos, K G; Giannouli, I M; Androutsopoulos, K N; Ziomas, I; Kosmidis, E; Simeonidis, P; Poupkou, N

2002-01-28

The most important characteristic of major chemical accidents, from a societal perspective, is their tendency to produce off-site effects. The extent and severity of the accident may significantly affect the population and the environment of the adjacent areas. Following an accident event, effort should be made to limit such effects. Management decisions should be based on rational and quantitative information based on the site specific circumstances and the possible consequences. To produce such information we have developed an operational centre for managing large-scale industrial accidents. Its architecture involves an integrated framework of geographical information system (GIS) and RDBMS technology systems equipped with interactive communication capabilities. The operational centre was developed for Windows 98 platforms, for the region of Thriasion Pedion of West Attica, where the concentration of industrial activity and storage of toxic chemical is immense within areas of high population density. An appropriate case study is given in order to illuminate the use and necessity of the operational centre.

Intelligent systems for the autonomous exploration of Titan and Enceladus

NASA Astrophysics Data System (ADS)

Furfaro, Roberto; Lunine, Jonathan I.; Kargel, Jeffrey S.; Fink, Wolfgang

2008-04-01

Future planetary exploration of the outer satellites of the Solar System will require higher levels of onboard automation, including autonomous determination of sites where the probability of significant scientific findings is highest. Generally, the level of needed automation is heavily influenced by the distance between Earth and the robotic explorer(s) (e.g. spacecraft(s), rover(s), and balloon(s)). Therefore, planning missions to the outer satellites mandates the analysis, design and integration within the mission architecture of semi- and/or completely autonomous intelligence systems. Such systems should (1) include software packages that enable fully automated and comprehensive identification, characterization, and quantification of feature information within an operational region with subsequent target prioritization and selection for close-up reexamination; and (2) integrate existing information with acquired, "in transit" spatial and temporal sensor data to automatically perform intelligent planetary reconnaissance, which includes identification of sites with the highest potential to yield significant geological and astrobiological information. In this paper we review and compare some of the available Artificial Intelligence (AI) schemes and their adaptation to the problem of designing expert systems for onboard-based, autonomous science to be performed in the course of outer satellites exploration. More specifically, the fuzzy-logic framework proposed is analyzed in some details to show the effectiveness of such a scheme when applied to the problem of designing expert systems capable of identifying and further exploring regions on Titan and/or Enceladus that have the highest potential to yield evidence for past or present life. Based on available information (e.g., Cassini data), the current knowledge and understanding of Titan and Enceladus environments is evaluated to define a path for the design of a fuzzy-based system capable of reasoning over collected data and capable of providing the inference required to autonomously optimize future outer satellites explorations.

Analysis of a fuel cell on-site integrated energy system for a residential complex

NASA Technical Reports Server (NTRS)

Simons, S. N.; Maag, W. L.

1979-01-01

Declining supplies of domestic oil and gas and the increased cost of energy resulted in a renewed emphasis in utilizing available resources in the most efficient manner possible. This, in turn, brought about a reassessment of a number of methods for converting fossil fuels to end uses at the highest practical efficiency. One of these is the on-site integrated energy system (OS/IES). This system provides electric power from an on-site power plant and recovers heat from the power plant that would normally be rejected to the environment. An OS/IES is potentially useful in any application that requires both electricity and heat. Several OS/IES are analyzed for a residential complex. The paper is divided into two sections; the first compares three energy supply systems, the second compares various designs for fuel cell OS/IES.

Loss of the Otx2-Binding Site in the Nanog Promoter Affects the Integrity of Embryonic Stem Cell Subtypes and Specification of Inner Cell Mass-Derived Epiblast.

PubMed

Acampora, Dario; Omodei, Daniela; Petrosino, Giuseppe; Garofalo, Arcomaria; Savarese, Marco; Nigro, Vincenzo; Di Giovannantonio, Luca Giovanni; Mercadante, Vincenzo; Simeone, Antonio

2016-06-21

Mouse embryonic stem cells (ESCs) and the inner cell mass (ICM)-derived epiblast exhibit naive pluripotency. ESC-derived epiblast stem cells (EpiSCs) and the postimplantation epiblast exhibit primed pluripotency. Although core pluripotency factors are well-characterized, additional regulators, including Otx2, recently have been shown to function during the transition from naive to primed pluripotency. Here we uncover a role for Otx2 in the control of the naive pluripotent state. We analyzed Otx2-binding activity in ESCs and EpiSCs and identified Nanog, Oct4, and Sox2 as direct targets. To unravel the Otx2 transcriptional network, we targeted the strongest Otx2-binding site in the Nanog promoter, finding that this site modulates the size of specific ESC-subtype compartments in cultured cells and promotes Nanog expression in vivo, predisposing ICM differentiation to epiblast. Otx2-mediated Nanog regulation thus contributes to the integrity of the ESC state and cell lineage specification in preimplantation development. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

Web Based Autonomous Geophysical/Hydrological Monitoring of the Gilt Edge Mine Site: Implementation and Results

NASA Astrophysics Data System (ADS)

Versteeg, R. J.; Wangerud, K.; Mattson, E.; Ankeny, M.; Richardson, A.; Heath, G.

2005-05-01

The Ruby Gulch repository at the Gilt Edge Mine Superfund site is a capped waste rock repository. Early in the system design EPA and its subcontractor, Bureau of Reclamation, recognized the need for long-term monitoring system to provide information on the repository behavior with the following objectives: 1 Provide information on the integrity of the newly constructed surface cover and diversion system 2 Continually assess the waste's hydrological and geochemical behavior, such that rational decisions can be made for the operation of this cover and liner system 3 Easily access of information pertaining to the system performance to stakeholders 4 Integration of a variety of data sources to produce information which could be used to enhance future cover designs. Through discussions between EPA, the Bureau of Reclamation and Idaho National Laboratory a long-term monitoring system was designed and implemented allowing EPA to meet these objectives. This system was designed to provide a cost effective way to deal with massive amounts of data and information, subject to the following specifications: 1 Data acquisition should occur autonomously and automatically, 2 Data management, processing and presentation should be automated as much as possible, 3 Users should be able to access all data and information remotely through a web browser. The INL long-term monitoring system integrates the data from a set of 522 electrodes resistivity electrodes consisting of 462 surface electrodes and 60 borehole electrodes (in 4 wells with 15 electrodes each), an outflow meter at the toe of the repository, an autonomous, remotely accessible weather station, and four wells (average depths of 250 feet) with thermocouples, pressure transducers and sampling ports for water and air. The monitoring system has currently been in operation for over a year, and has collected data continuously over this period. Results from this system have shown both the diurnal variation in rockmass behavior, movement of water through the waste (allowing estimated in residence time) and are leading to a comprehensive model of the repository behavior. Due to the sheer volume of data, a user driven interface allows users to create their own views of the different datasets.

Implementation of an integrated computerized prescriber order-entry system for chemotherapy in a multisite safety-net health system.

PubMed

Chung, Clement; Patel, Shital; Lee, Rosetta; Fu, Lily; Reilly, Sean; Ho, Tuyet; Lionetti, Jason; George, Michael D; Taylor, Pam

2018-03-15

The development of a computerized prescriber order-entry (CPOE) system for chemotherapy in a multisite safety-net health system and the challenges to its successful implementation are described. Before CPOE for chemotherapy was first implemented and embedded in the electronic medical record system of Harris Health System (HHS), pharmacy personnel relied on regimen-specific preprinted order sets. However, due to differences in practice styles and workflow logistics, the paper orders across the 3 facilities were mostly site specific, with varying clinical content. Many of these order sets had not been approved by the oncology subcommittee. In addition, disparities in clinical knowledge and lack of communication contributed to inconsistencies in order set development. Led by medical directors from medical oncology departments at the 3 facilities, pharmacy administrators, and information technology representatives, HHS committed resources to supporting the adoption and use of a CPOE system for chemotherapy. Five practical lessons of broad applicability have been learned: engagement of interprofessional stakeholders, optimization of workflow before CPOE implementation, requirement of verification tool for CPOE, consolidation of protocols, and commitment to ongoing training and support. Evaluation of the CPOE system demonstrated a systemwide reduction in medication errors by 75% ( p < 0.05). Satisfaction with the CPOE system varied among sites and was unchanged institutionwide 6 months after the CPOE implementation. The development and implementation of CPOE for chemotherapy at a multisite safety-net health system created opportunities to optimize patient care and reduce variations through interprofessional collaborations. Initial evaluation suggested that CPOE reduced the medication-order error rate and improved user satisfaction in 1 of 3 facilities. Copyright © 2018 by the American Society of Health-System Pharmacists, Inc. All rights reserved.

[CRISPR/Cas system for genome editing in pluripotent stem cells].

PubMed

Vasil'eva, E A; Melino, D; Barlev, N A

2015-01-01

Genome editing systems based on site-specific nucleases became very popular for genome editing in modern bioengineering. Human pluripotent stem cells provide a unique platform for genes function study, disease modeling, and drugs testing. Consequently, technology for fast, accurate and well controlled genome manipulation is required. CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRISPR-associated) system could be employed for these purposes. This system is based on site-specific programmable nuclease Cas9. Numerous advantages of the CRISPR/Cas system and its successful application to human stem cells provide wide opportunities for genome therapy and regeneration medicine. In this publication, we describe and compare the main genome editing systems based on site-specific programmable nucleases and discuss opportunities and perspectives of the CRISPR/Cas system for application to pluripotent stem cells.

Onco-Regulon: an integrated database and software suite for site specific targeting of transcription factors of cancer genes

PubMed Central

Tomar, Navneet; Mishra, Akhilesh; Mrinal, Nirotpal; Jayaram, B.

2016-01-01

Transcription factors (TFs) bind at multiple sites in the genome and regulate expression of many genes. Regulating TF binding in a gene specific manner remains a formidable challenge in drug discovery because the same binding motif may be present at multiple locations in the genome. Here, we present Onco-Regulon (http://www.scfbio-iitd.res.in/software/onco/NavSite/index.htm), an integrated database of regulatory motifs of cancer genes clubbed with Unique Sequence-Predictor (USP) a software suite that identifies unique sequences for each of these regulatory DNA motifs at the specified position in the genome. USP works by extending a given DNA motif, in 5′→3′, 3′ →5′ or both directions by adding one nucleotide at each step, and calculates the frequency of each extended motif in the genome by Frequency Counter programme. This step is iterated till the frequency of the extended motif becomes unity in the genome. Thus, for each given motif, we get three possible unique sequences. Closest Sequence Finder program predicts off-target drug binding in the genome. Inclusion of DNA-Protein structural information further makes Onco-Regulon a highly informative repository for gene specific drug development. We believe that Onco-Regulon will help researchers to design drugs which will bind to an exclusive site in the genome with no off-target effects, theoretically. Database URL: http://www.scfbio-iitd.res.in/software/onco/NavSite/index.htm PMID:27515825

A 2.4-GHz ISM RF and UWB hybrid RFID real-time locating system for industrial enterprise Internet of Things

NASA Astrophysics Data System (ADS)

Zhai, Chuanying; Zou, Zhuo; Zhou, Qin; Mao, Jia; Chen, Qiang; Tenhunen, Hannu; Zheng, Lirong; Xu, Lida

2017-07-01

This paper presents a 2.4-GHz radio frequency (RF) and ultra-wide bandwidth (UWB) hybrid real-time locating system (RTLS) for industrial enterprise Internet of Things (IoT). It employs asymmetric wireless link, that is, UWB radio is utilised for accurate positioning up to 10 cm in critical sites, whereas 2.4-GHz RF is used for tag control and coarse positioning in non-critical sites. The specified communication protocol and the adaptive tag synchronisation rate ensure reliable and deterministic access with a scalable system capacity and avoid unpredictable latency and additional energy consumption of retransmissions due to collisions. The tag, consisting of a commercial 2.4-GHz transceiver and a customised application-specific integrated circuit (ASIC) UWB transmitter (Tx), is able to achieve up to 3 years' battery life at 1600 tags per position update second with 1000 mAh battery in one cluster. The time difference of arrival (TDoA)-based positioning experiment at UWB radio is performed on the designed software-defined radio (SDR) platform.

Electromagnetic Fields Exposure Limits

DTIC Science & Technology

2018-01-01

analysis, synthesis, integration and validation of knowledge derived through the scientific method. In NATO, S&T is addressed using different...Panel • NMSG NATO Modelling and Simulation Group • SAS System Analysis and Studies Panel • SCI Systems Concepts and Integration Panel • SET... integrity or morphology. They later also failed to find a lack of direct DNA damage in human blood (strand breaks, alkali-labile sites, and incomplete

Multiplexed, High Density Electrophysiology with Nanofabricated Neural Probes

PubMed Central

Du, Jiangang; Blanche, Timothy J.; Harrison, Reid R.; Lester, Henry A.; Masmanidis, Sotiris C.

2011-01-01

Extracellular electrode arrays can reveal the neuronal network correlates of behavior with single-cell, single-spike, and sub-millisecond resolution. However, implantable electrodes are inherently invasive, and efforts to scale up the number and density of recording sites must compromise on device size in order to connect the electrodes. Here, we report on silicon-based neural probes employing nanofabricated, high-density electrical leads. Furthermore, we address the challenge of reading out multichannel data with an application-specific integrated circuit (ASIC) performing signal amplification, band-pass filtering, and multiplexing functions. We demonstrate high spatial resolution extracellular measurements with a fully integrated, low noise 64-channel system weighing just 330 mg. The on-chip multiplexers make possible recordings with substantially fewer external wires than the number of input channels. By combining nanofabricated probes with ASICs we have implemented a system for performing large-scale, high-density electrophysiology in small, freely behaving animals that is both minimally invasive and highly scalable. PMID:22022568

On-chip cooling by superlattice-based thin-film thermoelectrics.

PubMed

Chowdhury, Ihtesham; Prasher, Ravi; Lofgreen, Kelly; Chrysler, Gregory; Narasimhan, Sridhar; Mahajan, Ravi; Koester, David; Alley, Randall; Venkatasubramanian, Rama

2009-04-01

There is a significant need for site-specific and on-demand cooling in electronic, optoelectronic and bioanalytical devices, where cooling is currently achieved by the use of bulky and/or over-designed system-level solutions. Thermoelectric devices can address these limitations while also enabling energy-efficient solutions, and significant progress has been made in the development of nanostructured thermoelectric materials with enhanced figures-of-merit. However, fully functional practical thermoelectric coolers have not been made from these nanomaterials due to the enormous difficulties in integrating nanoscale materials into microscale devices and packaged macroscale systems. Here, we show the integration of thermoelectric coolers fabricated from nanostructured Bi2Te3-based thin-film superlattices into state-of-the-art electronic packages. We report cooling of as much as 15 degrees C at the targeted region on a silicon chip with a high ( approximately 1,300 W cm-2) heat flux. This is the first demonstration of viable chip-scale refrigeration technology and has the potential to enable a wide range of currently thermally limited applications.

Predicting the risk of cucurbit downy mildew in the eastern United States using an integrated aerobiological model

NASA Astrophysics Data System (ADS)

Neufeld, K. N.; Keinath, A. P.; Gugino, B. K.; McGrath, M. T.; Sikora, E. J.; Miller, S. A.; Ivey, M. L.; Langston, D. B.; Dutta, B.; Keever, T.; Sims, A.; Ojiambo, P. S.

2017-11-01

Cucurbit downy mildew caused by the obligate oomycete, Pseudoperonospora cubensis, is considered one of the most economically important diseases of cucurbits worldwide. In the continental United States, the pathogen overwinters in southern Florida and along the coast of the Gulf of Mexico. Outbreaks of the disease in northern states occur annually via long-distance aerial transport of sporangia from infected source fields. An integrated aerobiological modeling system has been developed to predict the risk of disease occurrence and to facilitate timely use of fungicides for disease management. The forecasting system, which combines information on known inoculum sources, long-distance atmospheric spore transport and spore deposition modules, was tested to determine its accuracy in predicting risk of disease outbreak. Rainwater samples at disease monitoring sites in Alabama, Georgia, Louisiana, New York, North Carolina, Ohio, Pennsylvania and South Carolina were collected weekly from planting to the first appearance of symptoms at the field sites during the 2013, 2014, and 2015 growing seasons. A conventional PCR assay with primers specific to P. cubensis was used to detect the presence of sporangia in rain water samples. Disease forecasts were monitored and recorded for each site after each rain event until initial disease symptoms appeared. The pathogen was detected in 38 of the 187 rainwater samples collected during the study period. The forecasting system correctly predicted the risk of disease outbreak based on the presence of sporangia or appearance of initial disease symptoms with an overall accuracy rate of 66 and 75%, respectively. In addition, the probability that the forecasting system correctly classified the presence or absence of disease was ≥ 73%. The true skill statistic calculated based on the appearance of disease symptoms in cucurbit field plantings ranged from 0.42 to 0.58, indicating that the disease forecasting system had an acceptable to good performance in predicting the risk of cucurbit downy mildew outbreak in the eastern United States.

Predicting the risk of cucurbit downy mildew in the eastern United States using an integrated aerobiological model.

PubMed

Neufeld, K N; Keinath, A P; Gugino, B K; McGrath, M T; Sikora, E J; Miller, S A; Ivey, M L; Langston, D B; Dutta, B; Keever, T; Sims, A; Ojiambo, P S

2018-04-01

Cucurbit downy mildew caused by the obligate oomycete, Pseudoperonospora cubensis, is considered one of the most economically important diseases of cucurbits worldwide. In the continental United States, the pathogen overwinters in southern Florida and along the coast of the Gulf of Mexico. Outbreaks of the disease in northern states occur annually via long-distance aerial transport of sporangia from infected source fields. An integrated aerobiological modeling system has been developed to predict the risk of disease occurrence and to facilitate timely use of fungicides for disease management. The forecasting system, which combines information on known inoculum sources, long-distance atmospheric spore transport and spore deposition modules, was tested to determine its accuracy in predicting risk of disease outbreak. Rainwater samples at disease monitoring sites in Alabama, Georgia, Louisiana, New York, North Carolina, Ohio, Pennsylvania and South Carolina were collected weekly from planting to the first appearance of symptoms at the field sites during the 2013, 2014, and 2015 growing seasons. A conventional PCR assay with primers specific to P. cubensis was used to detect the presence of sporangia in rain water samples. Disease forecasts were monitored and recorded for each site after each rain event until initial disease symptoms appeared. The pathogen was detected in 38 of the 187 rainwater samples collected during the study period. The forecasting system correctly predicted the risk of disease outbreak based on the presence of sporangia or appearance of initial disease symptoms with an overall accuracy rate of 66 and 75%, respectively. In addition, the probability that the forecasting system correctly classified the presence or absence of disease was ≥ 73%. The true skill statistic calculated based on the appearance of disease symptoms in cucurbit field plantings ranged from 0.42 to 0.58, indicating that the disease forecasting system had an acceptable to good performance in predicting the risk of cucurbit downy mildew outbreak in the eastern United States.

New ΦBT1 site-specific integrative vectors with neutral phenotype in Streptomyces.

PubMed

Gonzalez-Quiñonez, Nathaly; López-García, María Teresa; Yagüe, Paula; Rioseras, Beatriz; Pisciotta, Annalisa; Alduina, Rosa; Manteca, Ángel

2016-03-01

Integrative plasmids are one of the best options to introduce genes in low copy and in a stable form into bacteria. The ΦC31-derived plasmids constitute the most common integrative vectors used in Streptomyces. They integrate at different positions (attB and pseudo-attB sites) generating different mutations. The less common ΦBT1-derived vectors integrate at the unique attB site localized in the SCO4848 gene (S. coelicolor genome) or their orthologues in other streptomycetes. This work demonstrates that disruption of SCO4848 generates a delay in spore germination. SCO4848 is co-transcribed with SCO4849, and the spore germination phenotype is complemented by SCO4849. Plasmids pNG1-4 were created by modifying the ΦBT1 integrative vector pMS82 by introducing a copy of SCO4849 under the control of the promoter region of SCO4848. pNG2 and pNG4 also included a copy of the P ermE * in order to facilitate gene overexpression. pNG3 and pNG4 harboured a copy of the bla gene (ampicillin resistance) to facilitate selection in E. coli. pNG1-4 are the only integrative vectors designed to produce a neutral phenotype when they are integrated into the Streptomyces genome. The experimental approach developed in this work can be applied to create phenotypically neutral integrative plasmids in other bacteria.

Bacterial Production of Site Specific 13C Labeled Phenylalanine and Methodology for High Level Incorporation into Bacterially Expressed Recombinant Proteins

PubMed Central

Ramaraju, Bhargavi; McFeeters, Hana; Vogler, Bernhard; McFeeters, Robert L.

2016-01-01

Nuclear magnetic resonance spectroscopy studies of ever larger systems have benefited from many different forms of isotope labeling, in particular, site specific isotopic labeling. Site specific 13C labeling of methyl groups has become an established means of probing systems not amenable to traditional methodology. However useful, methyl reporter sites can be limited in number and/or location. Therefore, new complementary site specific isotope labeling strategies are valuable. Aromatic amino acids make excellent probes since they are often found at important interaction interfaces and play significant structural roles. Aromatic side chains have many of the same advantages as methyl containing amino acids including distinct 13C chemical shifts and multiple magnetically equivalent 1H positions. Herein we report economical bacterial production and one-step purification of phenylalanine with 13C incorporation at the Cα, Cγ and Cε positions, resulting in two isolated 1H-13C spin systems. We also present methodology to maximize incorporation of phenylalanine into recombinantly overexpressed proteins in bacteria and demonstrate compatibility with ILV-methyl labeling. Inexpensive, site specific isotope labeled phenylalanine adds another dimension to biomolecular NMR, opening new avenues of study. PMID:28028744

Optimization of the Implementation of Managed Aquifer Recharge - Effects of Aquifer Heterogeneity

NASA Astrophysics Data System (ADS)

Maliva, Robert; Missimer, Thomas; Kneppers, Angeline

2010-05-01

Managed aquifer recharge (MAR) has become a key component of integrated water resources management, especially in water scarce regions. MAR can serve the dual role of increasing the supply of available water and improving the quality of recharged water through natural attenuation processes. The performance of MAR systems is highly dependent upon site-specific hydrogeological conditions. Aquifer heterogeneity, such as the presence of high-permeability preferential flow zones and dual or even the so-called triple-porosity conditions, has been responsible for the under performance or failure of some MAR systems. Aquifer heterogeneity can result in much more rapid and unpredictable movement and mixing of recharged water and the bypassing of natural attenuation processes. A critical element of MAR projects is a detailed aquifer characterization and the development of groundwater flow and solute transport models at the appropriate spatial and temporal scales that accurately simulate local heterogeneous flow systems. Geochemical modeling based on high-quality, site-specific mineralogical and water chemistry data can also be used to predict the potential for adverse water-rock interactions such as the leaching of arsenic and trace metals into recharged water. Hydrogeological conditions that could lead to poor system performance should be identified early in the project development before the investment is made to construct a full-scale system. Hydrogeological conditions that have lead to poor MAR system performance are typically identifiable at the exploratory well stage of projects. Early detection of adverse hydrogeological conditions provides an opportunity to either abandon a likely under-performing project, select an alternative site with more favorable conditions, or modify the system design to be more compatible with local hydrogeology. Advanced borehole geophysical techniques and workflow software can allow for enhanced aquifer characterization and thus allow for more successful MAR implementation as a tool for improved water resources management.

Quantifying site-specific physical heterogeneity within an estuarine seascape

USGS Publications Warehouse

Kennedy, Cristina G.; Mather, Martha E.; Smith, Joseph M.

2017-01-01

Quantifying physical heterogeneity is essential for meaningful ecological research and effective resource management. Spatial patterns of multiple, co-occurring physical features are rarely quantified across a seascape because of methodological challenges. Here, we identified approaches that measured total site-specific heterogeneity, an often overlooked aspect of estuarine ecosystems. Specifically, we examined 23 metrics that quantified four types of common physical features: (1) river and creek confluences, (2) bathymetric variation including underwater drop-offs, (3) land features such as islands/sandbars, and (4) major underwater channel networks. Our research at 40 sites throughout Plum Island Estuary (PIE) provided solutions to two problems. The first problem was that individual metrics that measured heterogeneity of a single physical feature showed different regional patterns. We solved this first problem by combining multiple metrics for a single feature using a within-physical feature cluster analysis. With this approach, we identified sites with four different types of confluences and three different types of underwater drop-offs. The second problem was that when multiple physical features co-occurred, new patterns of total site-specific heterogeneity were created across the seascape. This pattern of total heterogeneity has potential ecological relevance to structure-oriented predators. To address this second problem, we identified sites with similar types of total physical heterogeneity using an across-physical feature cluster analysis. Then, we calculated an additive heterogeneity index, which integrated all physical features at a site. Finally, we tested if site-specific additive heterogeneity index values differed for across-physical feature clusters. In PIE, the sites with the highest additive heterogeneity index values were clustered together and corresponded to sites where a fish predator, adult striped bass (Morone saxatilis), aggregated in a related acoustic tracking study. In summary, we have shown general approaches to quantifying site-specific heterogeneity.

Assessing the potential for AAV vector genotoxicity in a murine model

PubMed Central

Li, Hojun; Malani, Nirav; Hamilton, Shari R.; Schlachterman, Alexander; Bussadori, Giulio; Edmonson, Shyrie E.; Shah, Rachel; Arruda, Valder R.; Mingozzi, Federico; Fraser Wright, J.; Bushman, Frederic D.

2011-01-01

Gene transfer using adeno-associated virus (AAV) vectors has great potential for treating human disease. Recently, questions have arisen about the safety of AAV vectors, specifically, whether integration of vector DNA in transduced cell genomes promotes tumor formation. This study addresses these questions with high-dose liver-directed AAV-mediated gene transfer in the adult mouse as a model (80 AAV-injected mice and 52 controls). After 18 months of follow-up, AAV-injected mice did not show a significantly higher rate of hepatocellular carcinoma compared with controls. Tumors in mice treated with AAV vectors did not have significantly different amounts of vector DNA compared with adjacent normal tissue. A novel high-throughput method for identifying AAV vector integration sites was developed and used to clone 1029 integrants. Integration patterns in tumor tissue and adjacent normal tissue were similar to each other, showing preferences for active genes, cytosine-phosphate-guanosine islands, and guanosine/cysteine-rich regions. Gene expression data showed that genes near integration sites did not show significant changes in expression patterns compared with genes more distal to integration sites. No integration events were identified as causing increased oncogene expression. Thus, we did not find evidence that AAV vectors cause insertional activation of oncogenes and subsequent tumor formation. PMID:21106988

Effect of Plasmid Design and Type of Integration Event on Recombinant Protein Expression in Pichia pastoris.

PubMed

Vogl, Thomas; Gebbie, Leigh; Palfreyman, Robin W; Speight, Robert

2018-03-15

Pichia pastoris (syn. Komagataella phaffii ) is one of the most common eukaryotic expression systems for heterologous protein production. Expression cassettes are typically integrated in the genome to obtain stable expression strains. In contrast to Saccharomyces cerevisiae , where short overhangs are sufficient to target highly specific integration, long overhangs are more efficient in P. pastoris and ectopic integration of foreign DNA can occur. Here, we aimed to elucidate the influence of ectopic integration by high-throughput screening of >700 transformants and whole-genome sequencing of 27 transformants. Different vector designs and linearization approaches were used to mimic the most common integration events targeted in P. pastoris Fluorescence of an enhanced green fluorescent protein (eGFP) reporter protein was highly uniform among transformants when the expression cassettes were correctly integrated in the targeted locus. Surprisingly, most nonspecifically integrated transformants showed highly uniform expression that was comparable to specific integration, suggesting that nonspecific integration does not necessarily influence expression. However, a few clones (<10%) harboring ectopically integrated cassettes showed a greater variation spanning a 25-fold range, surpassing specifically integrated reference strains up to 6-fold. High-expression strains showed a correlation between increased gene copy numbers and high reporter protein fluorescence levels. Our results suggest that for comparing expression levels between strains, the integration locus can be neglected as long as a sufficient numbers of transformed strains are compared. For expression optimization of highly expressible proteins, increasing copy number appears to be the dominant positive influence rather than the integration locus, genomic rearrangements, deletions, or single-nucleotide polymorphisms (SNPs). IMPORTANCE Yeasts are commonly used as biotechnological production hosts for proteins and metabolites. In the yeast Saccharomyces cerevisiae , expression cassettes carrying foreign genes integrate highly specifically at the targeted sites in the genome. In contrast, cassettes often integrate at random genomic positions in nonconventional yeasts, such as Pichia pastoris (syn. Komagataella phaffii ). Hence, cells from the same transformation event often behave differently, with significant clonal variation necessitating the screening of large numbers of strains. The importance of this study is that we systematically investigated the influence of integration events in more than 700 strains. Our findings provide novel insight into clonal variation in P. pastoris and, thus, how to avoid pitfalls and obtain reliable results. The underlying mechanisms may also play a role in other yeasts and hence could be generally relevant for recombinant yeast protein production strains. Copyright © 2018 American Society for Microbiology.

Optimization of Streptomyces bacteriophage phi C31 integrase system to prevent post integrative gene silencing in pulmonary type II cells.

PubMed

Aneja, Manish Kumar; Geiger, Johannes; Imker, Rabea; Uzgun, Senta; Kormann, Michael; Hasenpusch, Guenther; Maucksch, Christof; Rudolph, Carsten

2009-12-31

phi C31 integrase has emerged as a potent tool for achieving long-term gene expression in different tissues. The present study aimed at optimizing elements of phi C31 integrase system for alveolar type II cells. Luciferase and beta-galactosidase activities were measured at different time points post transfection. 5-Aza-2'deoxycytidine (AZA) and trichostatin A (TSA) were used to inhibit DNA methyltransferase and histone deacetylase complex (HDAC) respectively. In A549 cells, expression of the integrase using a CMV promoter resulted in highest integrase activity, whereas in MLE12 cells, both CAG and CMV promoter were equally effective. Effect of polyA site was observed only in A549 cells, where replacement of SV40 polyA by bovine growth hormone (BGH) polyA site resulted in an enhancement of integrase activity. Addition of a C-terminal SV40 nuclear localization signal (NLS) did not result in any significant increase in integrase activity. Long-term expression studies with AZA and TSA, provided evidence for post-integrative gene silencing. In MLE12 cells, both DNA methylases and HDACs played a significant role in silencing, whereas in A549 cells, it could be attributed majorly to HDAC activity. Donor plasmids comprising cellular promoters ubiquitin B (UBB), ubiquitin C (UCC) and elongation factor 1 alpha (EF1 alpha) in an improved backbone prevented post-integrative gene silencing. In contrast to A549 and MLE12 cells, no silencing could be observed in human bronchial epithelial cells, BEAS-2B. Donor plasmid coding for murine erythropoietin under the EF1 alpha promoter when combined with phi C31 integrase resulted in higher long-term erythropoietin expression and subsequently higher hematocrit levels in mice after intravenous delivery to the lungs. These results provide evidence for cell specific post integrative gene silencing with C31 integrase and demonstrate the pivotal role of donor plasmid in long-term expression attained with this system.

Improvements in algal lipid production: a systems biology and gene editing approach.

PubMed

Banerjee, Avik; Banerjee, Chiranjib; Negi, Sangeeta; Chang, Jo-Shu; Shukla, Pratyoosh

2018-05-01

In the wake of rising energy demands, microalgae have emerged as potential sources of sustainable and renewable carbon-neutral fuels, such as bio-hydrogen and bio-oil. For rational metabolic engineering, the elucidation of metabolic pathways in fine detail and their manipulation according to requirements is the key to exploiting the use of microalgae. Emergence of site-specific nucleases have revolutionized applied research leading to biotechnological gains. Genome engineering as well as modulation of the endogenous genome with high precision using CRISPR systems is being gradually employed in microalgal research. Further, to optimize and produce better algal platforms, use of systems biology network analysis and integration of omics data is required. This review discusses two important approaches: systems biology and gene editing strategies used on microalgal systems with a focus on biofuel production and sustainable solutions. It also emphasizes that the integration of such systems would contribute and compliment applied research on microalgae. Recent advances in microalgae are discussed, including systems biology, gene editing approaches in lipid bio-synthesis, and antenna engineering. Lastly, it has been attempted here to showcase how CRISPR/Cas systems are a better editing tool than existing techniques that can be utilized for gene modulation and engineering during biofuel production.

Intein-mediated site-specific synthesis of tumor-targeting protein delivery system: Turning PEG dilemma into prodrug-like feature

PubMed Central

Chen, Yingzhi; Zhang, Meng; Jin, Hongyue; Tang, Yisi; Wang, Huiyuan; Xu, Qin; Li, Yaping; Li, Feng; Huang, Yongzhuo

2017-01-01

Poor tumor-targeted and cytoplasmic delivery is a bottleneck for protein toxin-based cancer therapy. Ideally, a protein toxin drug should remain stealthy in circulation for prolonged half-life and reduced side toxicity, but turn activated at tumor. PEGylation is a solution to achieve the first goal, but creates a hurdle for the second because PEG rejects interaction between the drugs and tumor cells therein. Such PEG dilemma is an unsolved problem in protein delivery. Herein proposed is a concept of turning PEG dilemma into prodrug-like feature. A site-selectively PEGylated, gelatinase-triggered cell-penetrating trichosanthin protein delivery system is developed with three specific aims. The first is to develop an intein-based ligation method for achieving site-specific modification of protein toxins. The second is to develop a prodrug feature that renders protein toxins remaining stealthy in blood for reduced side toxicity and improved EPR effect. The third is to develop a gelatinase activatable cell-penetration strategy for enhanced tumor targeting and cytoplasmic delivery. Of note, site-specific modification is a big challenge in protein drug research, especially for such a complicated, multifunctional protein delivery system. We successfully develop a protocol for constructing a macromolecular prodrug system with intein-mediated ligation synthesis. With an on-column process of purification and intein-mediated cleavage, the site-specific PEGylation then can be readily achieved by conjugation with the activated C-terminus, thus constructing a PEG-capped, cell-penetrating trichosanthin system with a gelatinase-cleavable linker that enables tumor-specific activation of cytoplasmic delivery. It provides a promising method to address the PEG dilemma for enhanced protein drug delivery, and importantly, a facile protocol for site-specific modification of such a class of protein drugs for improving their druggability and industrial translation. PMID:27914267

Determinants that specify the integration pattern of retrotransposon Tf1 in the fbp1 promoter of Schizosaccharomyces pombe.

PubMed

Majumdar, Anasuya; Chatterjee, Atreyi Ghatak; Ripmaster, Tracy L; Levin, Henry L

2011-01-01

Long terminal repeat (LTR) retrotransposons are closely related to retroviruses and, as such, are important models for the study of viral integration and target site selection. The transposon Tf1 of Schizosaccharomyces pombe integrates with a strong preference for the promoters of polymerase II (Pol II)-transcribed genes. Previous work in vivo with plasmid-based targets revealed that the patterns of insertion were promoter specific and highly reproducible. To determine which features of promoters are recognized by Tf1, we studied integration in a promoter that has been characterized. The promoter of fbp1 has two upstream activating sequences, UAS1 and UAS2. We found that integration was targeted to two windows, one 180 nucleotides (nt) upstream and the other 30 to 40 nt downstream of UAS1. A series of deletions in the promoter showed that the integration activities of these two regions functioned autonomously. Integration assays of UAS2 and of a synthetic promoter demonstrated that strong promoter activity alone was not sufficient to direct integration. The factors that modulate the transcription activities of UAS1 and UAS2 include the activators Atf1p, Pcr1p, and Rst2p as well as the repressors Tup11p, Tup12p, and Pka1p. Strains lacking each of these proteins revealed that Atf1p alone mediated the sites of integration. These data indicate that Atf1p plays a direct and specific role in targeting integration in the promoter of fbp1.

Determinants That Specify the Integration Pattern of Retrotransposon Tf1 in the fbp1 Promoter of Schizosaccharomyces pombe ▿ †

PubMed Central

Majumdar, Anasuya; Chatterjee, Atreyi Ghatak; Ripmaster, Tracy L.; Levin, Henry L.

2011-01-01

Long terminal repeat (LTR) retrotransposons are closely related to retroviruses and, as such, are important models for the study of viral integration and target site selection. The transposon Tf1 of Schizosaccharomyces pombe integrates with a strong preference for the promoters of polymerase II (Pol II)-transcribed genes. Previous work in vivo with plasmid-based targets revealed that the patterns of insertion were promoter specific and highly reproducible. To determine which features of promoters are recognized by Tf1, we studied integration in a promoter that has been characterized. The promoter of fbp1 has two upstream activating sequences, UAS1 and UAS2. We found that integration was targeted to two windows, one 180 nucleotides (nt) upstream and the other 30 to 40 nt downstream of UAS1. A series of deletions in the promoter showed that the integration activities of these two regions functioned autonomously. Integration assays of UAS2 and of a synthetic promoter demonstrated that strong promoter activity alone was not sufficient to direct integration. The factors that modulate the transcription activities of UAS1 and UAS2 include the activators Atf1p, Pcr1p, and Rst2p as well as the repressors Tup11p, Tup12p, and Pka1p. Strains lacking each of these proteins revealed that Atf1p alone mediated the sites of integration. These data indicate that Atf1p plays a direct and specific role in targeting integration in the promoter of fbp1. PMID:20980525

Environmental endocrine disruption: an effects assessment and analysis.

PubMed Central

Crisp, T M; Clegg, E D; Cooper, R L; Wood, W P; Anderson, D G; Baetcke, K P; Hoffmann, J L; Morrow, M S; Rodier, D J; Schaeffer, J E; Touart, L W; Zeeman, M G; Patel, Y M

1998-01-01

This report is an overview of the current state of the science relative to environmental endocrine disruption in humans, laboratory testing, and wildlife species. Background information is presented on the field of endocrinology, the nature of hormones, and potential sites for endocrine disruption, with specific examples of chemicals affecting these sites. An attempt is made to present objectively the issue of endocrine disruption, consider working hypotheses, offer opposing viewpoints, analyze the available information, and provide a reasonable assessment of the problem. Emphasis is placed on disruption of central nervous system--pituitary integration of hormonal and sexual behavioral activity, female and male reproductive system development and function, and thyroid function. In addition, the potential role of environmental endocrine disruption in the induction of breast, testicular, and prostate cancers, as well as endometriosis, is evaluated. The interrelationship of the endocrine and immune system is documented. With respect to endocrine-related ecological effects, specific case examples from the peer-reviewed literature of marine invertebrates and representatives of the five classes of vertebrates are presented and discussed. The report identifies some data gaps in our understanding of the environmental endocrine disruption issue and recommends a few research needs. Finally, the report states the U.S. Environmental Protection Agency Science Policy Council's interim position on endocrine disruption and lists some of the ongoing activities to deal with this matter. PMID:9539004

Experiences from site-specific landslide early warning systems

NASA Astrophysics Data System (ADS)

Michoud, C.; Bazin, S.; Blikra, L. H.; Derron, M.-H.; Jaboyedoff, M.

2013-10-01

Landslide early warning systems (EWSs) have to be implemented in areas with large risk for populations or infrastructures when classical structural remediation measures cannot be set up. This paper aims to gather experiences of existing landslide EWSs, with a special focus on practical requirements (e.g., alarm threshold values have to take into account the smallest detectable signal levels of deployed sensors before being established) and specific issues when dealing with system implementations. Within the framework of the SafeLand European project, a questionnaire was sent to about one-hundred institutions in charge of landslide management. Finally, we interpreted answers from experts belonging to 14 operational units related to 23 monitored landslides. Although no standard requirements exist for designing and operating EWSs, this review highlights some key elements, such as the importance of pre-investigation work, the redundancy and robustness of monitoring systems, the establishment of different scenarios adapted to gradual increasing of alert levels, and the necessity of confidence and trust between local populations and scientists. Moreover, it also confirms the need to improve our capabilities for failure forecasting, monitoring techniques and integration of water processes into landslide conceptual models.

Application of Linear Array Imaging Techniques to the Real-Time Inspection of Airframe Structures and Substructures

NASA Technical Reports Server (NTRS)

Miller, James G. (Principal Investigator)

1996-01-01

Current concern for ensuring, the air-worthiness of the aging commercial air fleet has prompted the establishment of broad-agency programs to develop NDT technologies that address specific aging-aircraft issues. One of the crucial technological needs that has been identified is the development of rapid, quantitative systems for depot-level inspection of bonded aluminum lap joints on aircraft. Research results for characterization of disbond and corrosion based on normal-incidence pulse-echo measurement geometries are showing promise, but are limited by the single-site nature of the measurement which requires manual or mechanical scanning to inspect an area. One approach to developing efficient systems may be to transfer specific aspects of current medical imaging technology to the NDT arena. Ultrasonic medical imaging, systems offer many desirable attributes for large scale inspection. They are portable, provide real-time imaging, and have integrated video tape recorder and printer capabilities available for documentation and post-inspection review. Furthermore, these systems are available at a relatively low cost (approximately $50,000 to $200,000) and can be optimized for use with metals with straight-forward modifications.

Recombination between bacteriophage lambda and plasmid pBR322 in Escherichia coli.

PubMed Central

Pogue-Geile, K L; Dassarma, S; King, S R; Jaskunas, S R

1980-01-01

Recombinant lambda phages were isolated that resulted from recombination between the lambda genome and plasmid pBR322 in Escherichia coli, even though these deoxyribonucleic acids (DNAs) did not share extensive regions of homology. The characterization of these recombinant DNAs by heteroduplex analysis and restriction endonucleases is described. All but one of the recombinants appeared to have resulted from reciprocal recombination between a site on lambda DNA and a site on the plasmid. In general, there were two classes of recombinants. One class appeared to have resulted from recombination at the phage attachment site that probably resulted from lambda integration into secondary attachment sites on the plasmid. Seven different secondary attachment sites on pBR322 were found. The other class resulted from plasmid integration at other sites that were widely scattered on the lambda genome. For this second class of recombinants, more than one site on the plasmid could recombine with lambda DNA. Thus, the recombination did not appear to be site specific with respect to lambda or the plasmid. Possible mechanisms for generating these recombinants are discussed. Images PMID:6247334

Using Web-based Tutorials To Enhance Library Instruction.

ERIC Educational Resources Information Center

Kocour, Bruce G.

2000-01-01

Describes the development of a Web site for library instruction at Carson-Newman College (TN) and its integration into English composition courses. Describes the use of a virtual tour, a tutorial on database searching, tutorials on specific databases, and library guides to specific disciplines to create an effective mechanism for active learning.…

An Integrated Nutrition Education Program for Dental Students.

ERIC Educational Resources Information Center

DePaola, Dominick P.; And Others

1978-01-01

Presents a diagonal system of nutrition education in a dental program that integrates didactic, clinical work, and community sites. Discusses the three phases including educational methodology, evaluation strategy, and unique program features. (MA)

Integrated quality assessment of sediments from harbour areas in Santos-São Vicente Estuarine System, Southern Brazil

NASA Astrophysics Data System (ADS)

Buruaem, Lucas Moreira; de Castro, Ítalo Braga; Hortellani, Marcos Antonio; Taniguchi, Satie; Fillmann, Gilberto; Sasaki, Silvio Tarou; Varella Petti, Mônica Angélica; Sarkis, Jorge Eduardo de Souza; Bícego, Márcia Caruso; Maranho, Luciane Alves; Davanso, Marcela Bergo; Nonato, Edmundo Ferraz; Cesar, Augusto; Costa-Lotufo, Leticia Veras; Abessa, Denis Moledo de Souza

2013-09-01

Santos-São Vicente Estuarine System is a highly populated coastal zone in Brazil and where it is located the major port of Latin America. Historically, port activities, industrial and domestic effluents discharges have constituted the main sources of contaminants to estuarine system. This study aimed to assess the recent status of sediment quality from 5 zones of Port of Santos by applying a lines-of-evidence approach through integrating results of: (1) acute toxicity of whole sediment and chronic toxicity of liquid phases; (2) grain size, organic matter, organic carbon, nitrogen, phosphorus, trace metals, polycyclic aromatic hydrocarbons, linear alkylbenzenes and butyltins; (3) benthic community descriptors. Results revealed a gradient of increasing contamination for metals and organic compounds, alongside with their geochemical carriers. Sediment liquid phases were more toxic compared to whole sediment. Low number of species and individuals indicated the impoverishment of benthic community. The use of site-specific sediment quality guidelines was more appropriate to predict sediment toxicity. The integration of results through Sediment Quality Triad approach and principal component analysis allowed observing the effects of natural stressors and dredging on sediment quality and benthic distribution. Even with recent governmental efforts to control, pollution is still relevant in Port of Santos and a threat to local ecosystems.

Initial source and site characterization studies for the U.C. Santa Barbara campus

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

Archuleta, R.; Nicholson, C.; Steidl, J.

1997-12-01

The University of California Campus-Laboratory Collaboration (CLC) project is an integrated 3 year effort involving Lawrence Livermore National Laboratory (LLNL) and four UC campuses - Los Angeles (UCLA), Riverside (UCR), Santa Barbara (UCSB), and San Diego (UCSD) - plus additional collaborators at San Diego State University (SDSU), at Los Alamos National Laboratory and in industry. The primary purpose of the project is to estimate potential ground motions from large earthquakes and to predict site-specific ground motions for one critical structure on each campus. This project thus combines the disciplines of geology, seismology, geodesy, soil dynamics, and earthquake engineering into amore » fully integrated approach. Once completed, the CLC project will provide a template to evaluate other buildings at each of the four UC campuses, as well as provide a methodology for evaluating seismic hazards at other critical sites in California, including other UC locations at risk from large earthquakes. Another important objective of the CLC project is the education of students and other professional in the application of this integrated, multidisciplinary, state-of-the-art approach to the assessment of earthquake hazard. For each campus targeted by the CLC project, the seismic hazard study will consist of four phases: Phase I - Initial source and site characterization, Phase II - Drilling, logging, seismic monitoring, and laboratory dynamic soil testing, Phase III - Modeling of predicted site-specific earthquake ground motions, and Phase IV - Calculations of 3D building response. This report cover Phase I for the UCSB campus and incudes results up through March 1997.« less

40 CFR 141.720 - Inactivation toolbox components.

Code of Federal Regulations, 2013 CFR

2013-07-01

... between the indicated values: Log credit = (0.0397 × (1.09757)Temp) × CT. (c) Site-specific study. The... this section on a site-specific basis. The State must base this approval on a site-specific study a... in this table are applicable only to post-filter applications of UV in filtered systems and to...

40 CFR 141.720 - Inactivation toolbox components.

Code of Federal Regulations, 2010 CFR

2010-07-01

... between the indicated values: Log credit = (0.0397 × (1.09757)Temp) × CT. (c) Site-specific study. The... this section on a site-specific basis. The State must base this approval on a site-specific study a... in this table are applicable only to post-filter applications of UV in filtered systems and to...

40 CFR 141.720 - Inactivation toolbox components.

Code of Federal Regulations, 2011 CFR

2011-07-01

... between the indicated values: Log credit = (0.0397 × (1.09757)Temp) × CT. (c) Site-specific study. The... this section on a site-specific basis. The State must base this approval on a site-specific study a... in this table are applicable only to post-filter applications of UV in filtered systems and to...

40 CFR 141.720 - Inactivation toolbox components.

Code of Federal Regulations, 2012 CFR

2012-07-01

... between the indicated values: Log credit = (0.0397 × (1.09757)Temp) × CT. (c) Site-specific study. The... this section on a site-specific basis. The State must base this approval on a site-specific study a... in this table are applicable only to post-filter applications of UV in filtered systems and to...

Alteration of gene expression in human hepatocellular carcinoma with integrated hepatitis B virus DNA.

PubMed

Tamori, Akihiro; Yamanishi, Yoshihiro; Kawashima, Shuichi; Kanehisa, Minoru; Enomoto, Masaru; Tanaka, Hiromu; Kubo, Shoji; Shiomi, Susumu; Nishiguchi, Shuhei

2005-08-15

Integration of hepatitis B virus (HBV) DNA into the human genome is one of the most important steps in HBV-related carcinogenesis. This study attempted to find the link between HBV DNA, the adjoining cellular sequence, and altered gene expression in hepatocellular carcinoma (HCC) with integrated HBV DNA. We examined 15 cases of HCC infected with HBV by cassette ligation-mediated PCR. The human DNA adjacent to the integrated HBV DNA was sequenced. Protein coding sequences were searched for in the human sequence. In five cases with HBV DNA integration, from which good quality RNA was extracted, gene expression was examined by cDNA microarray analysis. The human DNA sequence successive to integrated HBV DNA was determined in the 15 HCCs. Eight protein-coding regions were involved: ras-responsive element binding protein 1, calmodulin 1, mixed lineage leukemia 2 (MLL2), FLJ333655, LOC220272, LOC255345, LOC220220, and LOC168991. The MLL2 gene was expressed in three cases with HBV DNA integrated into exon 3 of MLL2 and in one case with HBV DNA integrated into intron 3 of MLL2. Gene expression analysis suggested that two HCCs with HBV integrated into MLL2 had similar patterns of gene expression compared with three HCCs with HBV integrated into other loci of human chromosomes. HBV DNA was integrated at random sites of human DNA, and the MLL2 gene was one of the targets for integration. Our results suggest that HBV DNA might modulate human genes near integration sites, followed by integration site-specific expression of such genes during hepatocarcinogenesis.

An att site-based recombination reporter system for genome engineering and synthetic DNA assembly.

PubMed

Bland, Michael J; Ducos-Galand, Magaly; Val, Marie-Eve; Mazel, Didier

2017-07-14

Direct manipulation of the genome is a widespread technique for genetic studies and synthetic biology applications. The tyrosine and serine site-specific recombination systems of bacteriophages HK022 and ΦC31 are widely used for stable directional exchange and relocation of DNA sequences, making them valuable tools in these contexts. We have developed site-specific recombination tools that allow the direct selection of recombination events by embedding the attB site from each system within the β-lactamase resistance coding sequence (bla). The HK and ΦC31 tools were developed by placing the attB sites from each system into the signal peptide cleavage site coding sequence of bla. All possible open reading frames (ORFs) were inserted and tested for recombination efficiency and bla activity. Efficient recombination was observed for all tested ORFs (3 for HK, 6 for ΦC31) as shown through a cointegrate formation assay. The bla gene with the embedded attB site was functional for eight of the nine constructs tested. The HK/ΦC31 att-bla system offers a simple way to directly select recombination events, thus enhancing the use of site-specific recombination systems for carrying out precise, large-scale DNA manipulation, and adding useful tools to the genetics toolbox. We further show the power and flexibility of bla to be used as a reporter for recombination.

Neurite sprouting and synapse deterioration in the aging Caenorhabditis elegans nervous system.

PubMed

Toth, Marton Lorant; Melentijevic, Ilija; Shah, Leena; Bhatia, Aatish; Lu, Kevin; Talwar, Amish; Naji, Haaris; Ibanez-Ventoso, Carolina; Ghose, Piya; Jevince, Angela; Xue, Jian; Herndon, Laura A; Bhanot, Gyan; Rongo, Chris; Hall, David H; Driscoll, Monica

2012-06-27

Caenorhabditis elegans is a powerful model for analysis of the conserved mechanisms that modulate healthy aging. In the aging nematode nervous system, neuronal death and/or detectable loss of processes are not readily apparent, but because dendrite restructuring and loss of synaptic integrity are hypothesized to contribute to human brain decline and dysfunction, we combined fluorescence microscopy and electron microscopy (EM) to screen at high resolution for nervous system changes. We report two major components of morphological change in the aging C. elegans nervous system: (1) accumulation of novel outgrowths from specific neurons, and (2) physical decline in synaptic integrity. Novel outgrowth phenotypes, including branching from the main dendrite or new growth from somata, appear at a high frequency in some aging neurons, but not all. Mitochondria are often associated with age-associated branch sites. Lowered insulin signaling confers some maintenance of ALM and PLM neuron structural integrity into old age, and both DAF-16/FOXO and heat shock factor transcription factor HSF-1 exert neuroprotective functions. hsf-1 can act cell autonomously in this capacity. EM evaluation in synapse-rich regions reveals a striking decline in synaptic vesicle numbers and a diminution of presynaptic density size. Interestingly, old animals that maintain locomotory prowess exhibit less synaptic decline than same-age decrepit animals, suggesting that synaptic integrity correlates with locomotory healthspan. Our data reveal similarities between the aging C. elegans nervous system and mammalian brain, suggesting conserved neuronal responses to age. Dissection of neuronal aging mechanisms in C. elegans may thus influence the development of brain healthspan-extending therapies.

Neurite Sprouting and Synapse Deterioration in the Aging C. elegans Nervous System

PubMed Central

Toth, Marton; Melentijevic, Ilija; Shah, Leena; Bhatia, Aatish; Lu, Kevin; Talwar, Amish; Naji, Haaris; Ibanez-Ventoso, Carolina; Ghose, Piya; Jevince, Angela; Xue, Jian; Herndon, Laura A.; Bhanot, Gyan; Rongo, Chris; Hall, David H

2012-01-01

C. elegans is a powerful model for analysis of the conserved mechanisms that modulate healthy aging. In the aging nematode nervous system, neuronal death and/or detectable loss of processes are not readily apparent, but because dendrite restructuring and loss of synaptic integrity are hypothesized to contribute to human brain decline and dysfunction, we combined fluorescence microscopy and electron microscopy (EM) to screen at high resolution for nervous system changes. We report two major components of morphological change in the aging C. elegans nervous system: 1) accumulation of novel outgrowths from specific neurons, and 2) physical decline in synaptic integrity. Novel outgrowth phenotypes, including branching from the main dendrite or new growth from somata, appear at a high frequency in some aging neurons, but not all. Mitochondria are often associated with age-associated branch sites. Lowered insulin signaling confers some maintenance of ALM and PLM neuron structural integrity into old age, and both DAF-16/FOXO and heat shock factor transcription factor HSF-1 exert neuroprotective functions. hsf-1 can act cell autonomously in this capacity. EM evaluation in synapse-rich regions reveals a striking decline in synaptic vesicle numbers and a dimunition of presynaptic density size. Interestingly, old animals that maintain locomotory prowess exhibit less synaptic decline than same-age decrepit animals, suggesting that synaptic integrity correlates with locomotory healthspan. Our data reveal similarities between the aging C. elegans nervous system and mammalian brain, suggesting conserved neuronal responses to age. Dissection of neuronal aging mechanisms in C. elegans may thus influence the development of brain healthspan-extending therapies. PMID:22745480

Approaches for Enhancing Oral Bioavailability of Peptides and Proteins

PubMed Central

Renukuntla, Jwala; Vadlapudi, Aswani Dutt; Patel, Ashaben; Boddu, Sai HS.; Mitra, Ashim K

2013-01-01

Oral delivery of peptide and protein drugs faces immense challenge partially due to the gastrointestinal (GI) environment. In spite of considerable efforts by industrial and academic laboratories, no major breakthrough in the effective oral delivery of polypeptides and proteins has been accomplished. Upon oral administration, gastrointestinal epithelium acts as a physical and biochemical barrier for absorption of proteins resulting in low bioavailability (typically less than 1–2%). An ideal oral drug delivery system should be capable of a) maintaining the integrity of protein molecules until it reaches the site of absorption, b) releasing the drug at the target absorption site, where the delivery system appends to that site by virtue of specific interaction, and c) retaining inside the gastrointestinal tract irrespective of its transitory constraints. Various technologies have been explored to overcome the problems associated with the oral delivery of macromolecules such as insulin, gonadotropin-releasing hormones, calcitonin, human growth factor, vaccines, enkephalins, and interferons, all of which met with limited success. This review article intends to summarize the physiological barriers to oral delivery of peptides and proteins and novel pharmaceutical approaches to circumvent these barriers and enhance oral bioavailability of these macromolecules. PMID:23428883

Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions.

PubMed

Khatri, Kshitij; Klein, Joshua A; White, Mitchell R; Grant, Oliver C; Leymarie, Nancy; Woods, Robert J; Hartshorn, Kevan L; Zaia, Joseph

2016-06-01

Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. Mutations leading to changes in the number of HA glycosylation sites are often reported. Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Evaluation of potential water conservation using site-specific irrigation

USDA-ARS?s Scientific Manuscript database

With the advent of site-specific variable-rate irrigation (VRI) systems, irrigation can be spatially managed within sub-field-sized zones. Spatial irrigation management can optimize spatial water use efficiency and may conserve water. Spatial VRI systems are currently being managed by consultants ...

Environmental Management System Objectives & Targets Results Summary - FY 2015.

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

Vetter, Douglas W.

2016-02-01

Sandia National Laboratories (SNL) Environmental Management System is the integrated approach for members of the workforce to identify and manage environmental risks. Each Fiscal Year (FY) SNL performs an analysis to identify environmental aspects, and the environmental programs associated with them are charged with the task of routinely monitoring and measuring the objectives and targets that are established to mitigate potential impacts of SNL's operations on the environment. An annual summary of the results achieved towards meeting established Sandia Corporation and SNL Site-specific objectives and targets provides a connection to, and rational for, annually revised environmental aspects. The purpose ofmore » this document is to summarize the results achieved and documented in FY 2015.« less

Standards for Data Exchange and Case Management Information Systems in Support of Comprehensive Integrated School-Linked Services. Version 2.0.

ERIC Educational Resources Information Center

Far West Lab. for Educational Research and Development, San Francisco, CA.

This report is intended as a guide for local comprehensive integrated school-linked services sites and software vendors in developing and implementing case management information systems for the exchange and management of client data. The report is also intended to influence new development and future revisions of data systems, databases, and…

Genome-wide Determinants of Proviral Targeting, Clonal Abundance and Expression in Natural HTLV-1 Infection

PubMed Central

Melamed, Anat; Laydon, Daniel J.; Gillet, Nicolas A.; Tanaka, Yuetsu; Taylor, Graham P.; Bangham, Charles R. M.

2013-01-01

The regulation of proviral latency is a central problem in retrovirology. We postulate that the genomic integration site of human T lymphotropic virus type 1 (HTLV-1) determines the pattern of expression of the provirus, which in turn determines the abundance and pathogenic potential of infected T cell clones in vivo. We recently developed a high-throughput method for the genome-wide amplification, identification and quantification of proviral integration sites. Here, we used this protocol to test two hypotheses. First, that binding sites for transcription factors and chromatin remodelling factors in the genome flanking the proviral integration site of HTLV-1 are associated with integration targeting, spontaneous proviral expression, and in vivo clonal abundance. Second, that the transcriptional orientation of the HTLV-1 provirus relative to that of the nearest host gene determines spontaneous proviral expression and in vivo clonal abundance. Integration targeting was strongly associated with the presence of a binding site for specific host transcription factors, especially STAT1 and p53. The presence of the chromatin remodelling factors BRG1 and INI1 and certain host transcription factors either upstream or downstream of the provirus was associated respectively with silencing or spontaneous expression of the provirus. Cells expressing HTLV-1 Tax protein were significantly more frequent in clones of low abundance in vivo. We conclude that transcriptional interference and chromatin remodelling are critical determinants of proviral latency in natural HTLV-1 infection. PMID:23555266

Quantum kinetic expansion in the spin-boson model: Matrix formulation and system-bath factorized initial state.

PubMed

Gong, Zhihao; Tang, Zhoufei; Wang, Haobin; Wu, Jianlan

2017-12-28

Within the framework of the hierarchy equation of motion (HEOM), the quantum kinetic expansion (QKE) method of the spin-boson model is reformulated in the matrix representation. The equivalence between the two formulations (HEOM matrices and quantum operators) is numerically verified from the calculation of the time-integrated QKE rates. The matrix formulation of the QKE is extended to the system-bath factorized initial state. Following a one-to-one mapping between HEOM matrices and quantum operators, a quantum kinetic equation is rederived. The rate kernel is modified by an extra term following a systematic expansion over the site-site coupling. This modified QKE is numerically tested for its reliability by calculating the time-integrated rate and non-Markovian population kinetics. For an intermediate-to-strong dissipation strength and a large site-site coupling, the population transfer is found to be significantly different when the initial condition is changed from the local equilibrium to system-bath factorized state.

Plasmid partition system of the P1par family from the pWR100 virulence plasmid of Shigella flexneri.

PubMed

Sergueev, Kirill; Dabrazhynetskaya, Alena; Austin, Stuart

2005-05-01

P1par family members promote the active segregation of a variety of plasmids and plasmid prophages in gram-negative bacteria. Each has genes for ParA and ParB proteins, followed by a parS partition site. The large virulence plasmid pWR100 of Shigella flexneri contains a new P1par family member: pWR100par. Although typical parA and parB genes are present, the putative pWR100parS site is atypical in sequence and organization. However, pWR100parS promoted accurate plasmid partition in Escherichia coli when the pWR100 Par proteins were supplied. Unique BoxB hexamer motifs within parS define species specificities among previously described family members. Although substantially different from P1parS from the P1 plasmid prophage of E. coli, pWR100parS has the same BoxB sequence. As predicted, the species specificity of the two types proved identical. They also shared partition-mediated incompatibility, consistent with the proposed mechanistic link between incompatibility and species specificity. Among several informative sequence differences between pWR100parS and P1parS is the presence of a 21-bp insert at the center of the pWR100parS site. Deletion of this insert left much of the parS activity intact. Tolerance of central inserts with integral numbers of helical DNA turns reflects the critical topology of these sites, which are bent by binding the host IHF protein.

Integrated Assessment by the People: Insights from AgMIP Regional Teams in Sub-Saharan Africa and South Asia

NASA Astrophysics Data System (ADS)

Antle, J. M.

2017-12-01

AgMIP has developed innovative protocol-based methods for regional integrated assessment (RIA) that can be implemented by national researchers working with local and national stakeholders (http://www.agmip.org/regional-integrated-assessments-handbook/). The approach has been implemented by regional teams in Sub-Saharan Africa and South Asia. This presentation first summarizes novel elements of the AgMIP RIA methods, and their strengths and limitations, based on their application by AgMIP researchers. Key insights from the application of these methods for climate impact and adaptation in Sub-Saharan Africa and South Asia are presented. A major finding is that detailed, site-specific, systems-based analysis show much more local and regional variation in impacts than studies based on analysis of individual crops, and provide the basis for analysis of multi-faceted technology and policy options to facilitate the transition to sustainable and resilient development pathways. The presentation concludes with observations about advancing integrated assessments carried out by and for national and local researchers and stakeholders.

EPA Facility Registry Service (FRS): Facility Interests Dataset - Intranet

EPA Pesticide Factsheets

This web feature service consists of location and facility identification information from EPA's Facility Registry Service (FRS) for all sites that are available in the FRS individual feature layers. The layers comprise the FRS major program databases, including:Assessment Cleanup and Redevelopment Exchange System (ACRES) : brownfields sites ; Air Facility System (AFS) : stationary sources of air pollution ; Air Quality System (AQS) : ambient air pollution data from monitoring stations; Bureau of Indian Affairs (BIA) : schools data on Indian land; Base Realignment and Closure (BRAC) facilities; Clean Air Markets Division Business System (CAMDBS) : market-based air pollution control programs; Comprehensive Environmental Response, Compensation, and Liability Information System (CERCLIS) : hazardous waste sites; Integrated Compliance Information System (ICIS) : integrated enforcement and compliance information; National Compliance Database (NCDB) : Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and the Toxic Substances Control Act (TSCA); National Pollutant Discharge Elimination System (NPDES) module of ICIS : NPDES surface water permits; Radiation Information Database (RADINFO) : radiation and radioactivity facilities; RACT/BACT/LAER Clearinghouse (RBLC) : best available air pollution technology requirements; Resource Conservation and Recovery Act Information System (RCRAInfo) : tracks generators, transporters, treaters, storers, and disposers of haz

EPA Facility Registry Service (FRS): Facility Interests Dataset - Intranet Download

EPA Pesticide Factsheets

This downloadable data package consists of location and facility identification information from EPA's Facility Registry Service (FRS) for all sites that are available in the FRS individual feature layers. The layers comprise the FRS major program databases, including:Assessment Cleanup and Redevelopment Exchange System (ACRES) : brownfields sites ; Air Facility System (AFS) : stationary sources of air pollution ; Air Quality System (AQS) : ambient air pollution data from monitoring stations; Bureau of Indian Affairs (BIA) : schools data on Indian land; Base Realignment and Closure (BRAC) facilities; Clean Air Markets Division Business System (CAMDBS) : market-based air pollution control programs; Comprehensive Environmental Response, Compensation, and Liability Information System (CERCLIS) : hazardous waste sites; Integrated Compliance Information System (ICIS) : integrated enforcement and compliance information; National Compliance Database (NCDB) : Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and the Toxic Substances Control Act (TSCA); National Pollutant Discharge Elimination System (NPDES) module of ICIS : NPDES surface water permits; Radiation Information Database (RADINFO) : radiation and radioactivity facilities; RACT/BACT/LAER Clearinghouse (RBLC) : best available air pollution technology requirements; Resource Conservation and Recovery Act Information System (RCRAInfo) : tracks generators, transporters, treaters, storers, and disposers

EPA Facility Registry Service (FRS): Facility Interests Dataset Download

EPA Pesticide Factsheets

This downloadable data package consists of location and facility identification information from EPA's Facility Registry Service (FRS) for all sites that are available in the FRS individual feature layers. The layers comprise the FRS major program databases, including:Assessment Cleanup and Redevelopment Exchange System (ACRES) : brownfields sites ; Air Facility System (AFS) : stationary sources of air pollution ; Air Quality System (AQS) : ambient air pollution data from monitoring stations; Bureau of Indian Affairs (BIA) : schools data on Indian land; Base Realignment and Closure (BRAC) facilities; Clean Air Markets Division Business System (CAMDBS) : market-based air pollution control programs; Comprehensive Environmental Response, Compensation, and Liability Information System (CERCLIS) : hazardous waste sites; Integrated Compliance Information System (ICIS) : integrated enforcement and compliance information; National Compliance Database (NCDB) : Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and the Toxic Substances Control Act (TSCA); National Pollutant Discharge Elimination System (NPDES) module of ICIS : NPDES surface water permits; Radiation Information Database (RADINFO) : radiation and radioactivity facilities; RACT/BACT/LAER Clearinghouse (RBLC) : best available air pollution technology requirements; Resource Conservation and Recovery Act Information System (RCRAInfo) : tracks generators, transporters, treaters, storers, and disposers

EPA Facility Registry Service (FRS): Facility Interests Dataset

EPA Pesticide Factsheets

This web feature service consists of location and facility identification information from EPA's Facility Registry Service (FRS) for all sites that are available in the FRS individual feature layers. The layers comprise the FRS major program databases, including:Assessment Cleanup and Redevelopment Exchange System (ACRES) : brownfields sites ; Air Facility System (AFS) : stationary sources of air pollution ; Air Quality System (AQS) : ambient air pollution data from monitoring stations; Bureau of Indian Affairs (BIA) : schools data on Indian land; Base Realignment and Closure (BRAC) facilities; Clean Air Markets Division Business System (CAMDBS) : market-based air pollution control programs; Comprehensive Environmental Response, Compensation, and Liability Information System (CERCLIS) : hazardous waste sites; Integrated Compliance Information System (ICIS) : integrated enforcement and compliance information; National Compliance Database (NCDB) : Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and the Toxic Substances Control Act (TSCA); National Pollutant Discharge Elimination System (NPDES) module of ICIS : NPDES surface water permits; Radiation Information Database (RADINFO) : radiation and radioactivity facilities; RACT/BACT/LAER Clearinghouse (RBLC) : best available air pollution technology requirements; Resource Conservation and Recovery Act Information System (RCRAInfo) : tracks generators, transporters, treaters, storers, and disposers of haz

Solution of the classical Yang-Baxter equation with an exotic symmetry, and integrability of a multi-species boson tunnelling model

NASA Astrophysics Data System (ADS)

Links, Jon

2017-03-01

Solutions of the classical Yang-Baxter equation provide a systematic method to construct integrable quantum systems in an algebraic manner. A Lie algebra can be associated with any solution of the classical Yang-Baxter equation, from which commuting transfer matrices may be constructed. This procedure is reviewed, specifically for solutions without skew-symmetry. A particular solution with an exotic symmetry is identified, which is not obtained as a limiting expansion of the usual Yang-Baxter equation. This solution facilitates the construction of commuting transfer matrices which will be used to establish the integrability of a multi-species boson tunnelling model. The model generalises the well-known two-site Bose-Hubbard model, to which it reduces in the one-species limit. Due to the lack of an apparent reference state, application of the algebraic Bethe Ansatz to solve the model is prohibitive. Instead, the Bethe Ansatz solution is obtained by the use of operator identities and tensor product decompositions.

Opportunities for Launch Site Integrated System Health Engineering and Management

NASA Technical Reports Server (NTRS)

Waterman, Robert D.; Langwost, Patricia E.; Waterman, Susan J.

2005-01-01

The launch site processing flow involves operations such as functional verification, preflight servicing and launch. These operations often include hazards that must be controlled to protect human life and critical space hardware assets. Existing command and control capabilities are limited to simple limit checking durig automated monitoring. Contingency actions are highly dependent on human recognition, decision making, and execution. Many opportunities for Integrated System Health Engineering and Management (ISHEM) exist throughout the processing flow. This paper will present the current human-centered approach to health management as performed today for the shuttle and space station programs. In addition, it will address some of the more critical ISHEM needs, and provide recommendations for future implementation of ISHEM at the launch site.

VHF Air/Ground Communications for Air Traffic Control: A Decision Tree Approach to System Innovations. Volume 2

DTIC Science & Technology

1993-07-01

GROUND RADIO ISSUES Multiple transmitters and receivers at a common site has been a dfficult integration issue for many communicaton systems...lower data rate vocoders become available. 10. Compatibility of modulation with channel availability and message integrity improvements, including...overrides can use in- frequency and path diversity, integrity improvements band signaling, busy channel overrides, and Frequency diversity and ARQ ARQ is

The Integral Theory System Questionnaire: an anatomically directed questionnaire to determine pelvic floor dysfunctions in women.

PubMed

Wagenlehner, Florian Martin Erich; Fröhlich, Oliver; Bschleipfer, Thomas; Weidner, Wolfgang; Perletti, Gianpaolo

2014-06-01

Anatomical damage to pelvic floor structures may cause multiple symptoms. The Integral Theory System Questionnaire (ITSQ) is a holistic questionnaire that uses symptoms to help locate damage in specific connective tissue structures as a guide to reconstructive surgery. It is based on the integral theory, which states that pelvic floor symptoms and prolapse are both caused by lax suspensory ligaments. The aim of the present study was to psychometrically validate the ITSQ. Established psychometric properties including validity, reliability, and responsiveness were considered for evaluation. Criterion validity was assessed in a cohort of 110 women with pelvic floor dysfunctions by analyzing the correlation of questionnaire responses with objective clinical data. Test-retest was performed with questionnaires from 47 patients. Cronbach's alpha and "split-half" reliability coefficients were calculated for inner consistency analysis. Psychometric properties of ITSQ were comparable to the ones of previously validated Pelvic Floor Questionnaires. Face validity and content validity were approved by an expert group of the International Collaboration of Pelvic Floor surgeons. Convergent validity assessed using Bayesian method was at least as accurate as the expert assessment of anatomical defects. Objective data measurement in patients demonstrated significant correlations with ITSQ domains fulfilling criterion validity. Internal consistency values ranked from 0.85 to 0.89 in different scenarios. The ITSQ proofed accurate and is able to serve as a holistic Pelvic Floor Questionnaire directing symptoms to site-specific pelvic floor reconstructive surgery.

Integrated Smartphone-App-Chip System for On-Site Parts-Per-Billion-Level Colorimetric Quantitation of Aflatoxins.

PubMed

Li, Xiaochun; Yang, Fan; Wong, Jessica X H; Yu, Hua-Zhong

2017-09-05

We demonstrate herein an integrated, smartphone-app-chip (SPAC) system for on-site quantitation of food toxins, as demonstrated with aflatoxin B1 (AFB1), at parts-per-billion (ppb) level in food products. The detection is based on an indirect competitive immunoassay fabricated on a transparent plastic chip with the assistance of a microfluidic channel plate. A 3D-printed optical accessory attached to a smartphone is adapted to align the assay chip and to provide uniform illumination for imaging, with which high-quality images of the assay chip are captured by the smartphone camera and directly processed using a custom-developed Android app. The performance of this smartphone-based detection system was tested using both spiked and moldy corn samples; consistent results with conventional enzyme-linked immunosorbent assay (ELISA) kits were obtained. The achieved detection limit (3 ± 1 ppb, equivalent to μg/kg) and dynamic response range (0.5-250 ppb) meet the requested testing standards set by authorities in China and North America. We envision that the integrated SPAC system promises to be a simple and accurate method of food toxin quantitation, bringing much benefit for rapid on-site screening.

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

Jung, Haeryong; Lee, Eunyong; Jeong, YiYeong

Korea Radioactive-waste Management Corporation (KRMC) established in 2009 has started a new project to collect information on long-term stability of deep geological environments on the Korean Peninsula. The information has been built up in the integrated natural barrier database system available on web (www.deepgeodisposal.kr). The database system also includes socially and economically important information, such as land use, mining area, natural conservation area, population density, and industrial complex, because some of this information is used as exclusionary criteria during the site selection process for a deep geological repository for safe and secure containment and isolation of spent nuclear fuel andmore » other long-lived radioactive waste in Korea. Although the official site selection process has not been started yet in Korea, current integrated natural barrier database system and socio-economic database is believed that the database system will be effectively utilized to narrow down the number of sites where future investigation is most promising in the site selection process for a deep geological repository and to enhance public acceptance by providing readily-available relevant scientific information on deep geological environments in Korea. (authors)« less

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene - methyltetrazine reactions.

PubMed

Muraki, Michiro; Hirota, Kiyonori

2017-07-03

Fas ligand plays a key role in the human immune system as a major cell death inducing protein. The extracellular domain of human Fas ligand (hFasLECD) triggers apoptosis of malignant cells, and therefore is expected to have substantial potentials in medical biotechnology. However, the current application of this protein to clinical medicine is hampered by a shortage of the benefits relative to the drawbacks including the side-effects in systemic administration. Effective procedures for the engineering of the protein by attaching useful additional functions are required to overcome the problem. A procedure for the site-specific chemical conjugation of hFasLECD with a fluorochrome and functional proteins was devised using an inverse-electron-demand Diels-Alder reaction between trans-cyclooctene group and methyltetrazine group. The conjugations in the present study were attained by using much less molar excess amounts of the compounds to be attached as compared with the conventional chemical modification reactions using maleimide derivatives in the previous study. The isolated conjugates of hFasLECD with sulfo-Cy3, avidin and rabbit IgG Fab' domain presented the functional and the structural integrities of the attached molecules without impairing the specific binding activity toward human Fas receptor extracellular domain. The present study provided a new fundamental strategy for the production of the engineered hFasLECDs with additional beneficial functions, which will lead to the developments of the improved diagnostic systems and the effective treatment methods of serious diseases by using this protein as a component of novel molecular tools.

Integral equations in the study of polar and ionic interaction site fluids

PubMed Central

Howard, Jesse J.

2011-01-01

In this review article we consider some of the current integral equation approaches and application to model polar liquid mixtures. We consider the use of multidimensional integral equations and in particular progress on the theory and applications of three dimensional integral equations. The IEs we consider may be derived from equilibrium statistical mechanical expressions incorporating a classical Hamiltonian description of the system. We give example including salt solutions, inhomogeneous solutions and systems including proteins and nucleic acids. PMID:22383857

The Global GNSS, SLR, VLBI, and DORIS Networks and their Support of GGOS: IGS+ILRS+IVS+IDS

NASA Technical Reports Server (NTRS)

Noll, Carey

2008-01-01

The global network of the International GNSS Service (IGS), the International Laser Ranging Service (ILRS), the International VLBI Service for Geodesy and Astrometry (IVS), and the International DORIS Service (IDS) are part of the ground-based infrastructure for GGOS. The observations obtained from these global networks provide for the determination and maintenance of the International Terrestrial Reference Frame (ITRF), an accurate set of positions and velocities that provides a stable coordinate system allowing scientists ts to link measurements over space and time. Many of these sites offer co-location of two or more techniques. Co-location provides integration of technique-specific networks into the ITRF as well as an assessment/validation of the quality and accuracy of the resulting measurements. As of fall 2008, these networks consisted of 410 GNSS sites, 42 laser ranging sites, 45 VLBI sites, and 58 DORIS sites. This poster will illustrate the global coverage of these networks, highlighting inter-technique co-locations, and show the importance of these networks 60 the underlying goals of GGOS including providing the observational basis to maintain a stable, accurate, global reference frame.

Building place-based collaborations to develop high school students' groundwater systems knowledge and decision-making capacity

NASA Astrophysics Data System (ADS)

Podrasky, A.; Covitt, B. A.; Woessner, W.

2017-12-01

The availability of clean water to support human uses and ecological integrity has become an urgent interest for many scientists, decision makers and citizens. Likewise, as computational capabilities increasingly revolutionize and become integral to the practice of science, technology, engineering and math (STEM) disciplines, the STEM+ Computing (STEM+C) Partnerships program seeks to integrate the use of computational approaches in K-12 STEM teaching and learning. The Comp Hydro project, funded by a STEM+C grant from the National Science Foundation, brings together a diverse team of scientists, educators, professionals and citizens at sites in Arizona, Colorado, Maryland and Montana to foster water literacy, as well as computational science literacy, by integrating authentic, place- and data- based learning using physical, mathematical, computational and conceptual models. This multi-state project is currently engaging four teams of six teachers who work during two academic years with educators and scientists at each site. Teams work to develop instructional units specific to their region that integrate hydrologic science and computational modeling. The units, currently being piloted in high school earth and environmental science classes, provide a classroom context to investigate student understanding of how computation is used in Earth systems science. To develop effective science instruction that is rich in place- and data- based learning, effective collaborations between researchers, educators, scientists, professionals and citizens are crucial. In this poster, we focus on project implementation in Montana, where an instructional unit has been developed and is being tested through collaboration among University scientists, researchers and educators, high school teachers and agency and industry scientists and engineers. In particular, we discuss three characteristics of effective collaborative science education design for developing and implementing place- and data- based science education to support students in developing socio-scientific and computational literacy sufficient for making decisions about real world issues such as groundwater contamination. These characteristics include that science education experiences are real, responsive/accessible and rigorous.

The North Warning System: A Canadian military SATCOM success

NASA Astrophysics Data System (ADS)

Wawryk, Ivan J.

The strategic threat to North America presented by modern, supersonic bombers and long range cruise missiles has stimulated a modernization of atmospheric defence capability. The North Warning System (NWS), employs a chain of microwave radars across the Arctic and down the east coast of Canada to provide tactical warning and attack assessment information to NORAD and its forces. The responsibility for NWS facilities construction and communications is allocated to Canada. Earlier investigations in Canada and the U.S. had confirmed that an all satellite communications system to link the 47 Canadian NWS radars to command and control centers was more economical than any terrestrial alternative. A project was undertaken by the Canadian Department of National Defence (DND) to acquire and install an integrated, digital satellite communications system for the NWS. This paper outlines the project background and procurement process. System requirements and specifications are discussed and the communications system is described in some detail. The communications equipment will be required to function unmanned in extreme Arctic conditions for extended periods. Specifications called for a long haul communications network (LHCN), a maintenance control system to monitor and control the equipment, and a suite of on-site communications for each location. The LHCN is a fully integrated, redundant satellite system which employs a transponder on each of the Anik D satellites. Either transponder can carry the full communications load. The system is configured as a star network with the hub at North Bay. Five remote stations and the North Bay facility have been built in phase one of the project; the second phase will see the construction of 36 unattended stations across the Arctic and down the Baffin Island and Labrador coasts.

Diet and the anti-inflammatory effect of heat shock proteins.

PubMed

van Eden, Willem

2015-01-01

Stress proteins or heat shock proteins (HSPs) have a critical role in gut health and immune regulation. They have a functional significance as molecular chaperones for cell skeleton proteins and intercellular tight junction proteins. Herewith HSPs ensure gut epithelium integrity and effective intestinal barrier function. In addition, stress protein molecules such as HSP70 are a target for anti-inflammatory regulatory T cells (Tregs). Inflamed sites in the body feature inflammatory-stress induced enhanced levels of HSPs, which enable the immune system to target Tregs selectively to sites of inflammation. We have shown in experimental models of inflammatory diseases that both microbial HSP and endogenous (self) HSP molecules are capable of inducing the expansion of disease suppressive Tregs. Since the gut associated lymphoid tissue (GALT) is well poised towards the induction of regulation and tolerance, we set out to promote HSP expression and induction of Tregs in the gut lymphoid tissues by the oral administration of HSP co-inducing compounds. For the identification, selection and characterization of such compounds we have developed assay systems, such as reporter cell-lines, HSP specific T cell hybridomas and a transgenic mouse model (expression a HSP specific T cell receptor). The introduction of HSP coinducers into the diet constitutes a novel food based preventive or possibly even therapeutic approach in inflammatory diseases.

Comparative study of large scale simulation of underground explosions inalluvium and in fractured granite using stochastic characterization

NASA Astrophysics Data System (ADS)

Vorobiev, O.; Ezzedine, S. M.; Antoun, T.; Glenn, L.

2014-12-01

This work describes a methodology used for large scale modeling of wave propagation fromunderground explosions conducted at the Nevada Test Site (NTS) in two different geological settings:fractured granitic rock mass and in alluvium deposition. We show that the discrete nature of rockmasses as well as the spatial variability of the fabric of alluvium is very important to understand groundmotions induced by underground explosions. In order to build a credible conceptual model of thesubsurface we integrated the geological, geomechanical and geophysical characterizations conductedduring recent test at the NTS as well as historical data from the characterization during the undergroundnuclear test conducted at the NTS. Because detailed site characterization is limited, expensive and, insome instances, impossible we have numerically investigated the effects of the characterization gaps onthe overall response of the system. We performed several computational studies to identify the keyimportant geologic features specific to fractured media mainly the joints; and those specific foralluvium porous media mainly the spatial variability of geological alluvium facies characterized bytheir variances and their integral scales. We have also explored common key features to both geologicalenvironments such as saturation and topography and assess which characteristics affect the most theground motion in the near-field and in the far-field. Stochastic representation of these features based onthe field characterizations have been implemented in Geodyn and GeodynL hydrocodes. Both codeswere used to guide site characterization efforts in order to provide the essential data to the modelingcommunity. We validate our computational results by comparing the measured and computed groundmotion at various ranges. This work performed under the auspices of the U.S. Department of Energy by Lawrence LivermoreNational Laboratory under Contract DE-AC52-07NA27344.

The N-terminal region of the dopamine D2 receptor, a rhodopsin-like GPCR, regulates correct integration into the plasma membrane and endocytic routes

PubMed Central

Cho, DI; Min, C; Jung, KS; Cheong, SY; Zheng, M; Cheong, SJ; Oak, MH; Cheong, JH; Lee, BK; Kim, KM

2012-01-01

BACKGROUND AND PURPOSE Functional roles of the N-terminal region of rhodopsin-like GPCR family remain unclear. Using dopamine D2 and D3 receptors as a model system, we probed the roles of the N-terminal region in the signalling, intracellular trafficking of receptor proteins, and explored the critical factors that determine the functionality of the N-terminal region. EXPERIMENTAL APPROACH The N-terminal region of the D2 receptor was gradually shortened or switched with that of the D3 receptor or a non-specific sequence (FLAG), or potential N-terminal glycosylation sites were mutated. Effects of these manipulations on surface expression, internalization, post-endocytic behaviours and signalling were determined. KEY RESULTS Shortening the N-terminal region of the D2 receptor enhanced receptor internalization and impaired surface expression and signalling; ligand binding, desensitization and down-regulation were not affected but their association with a particular microdomain, caveolae, was disrupted. Replacement of critical residues within the N-terminal region with the FLAG epitope failed to restore surface expression but partially restored the altered internalization and signalling. When the N-terminal regions were switched between D2 and D3 receptors, cell surface expression pattern of each receptor was switched. Mutations of potential N-terminal glycosylation sites inhibited surface expression but enhanced internalization of D2 receptors. CONCLUSIONS AND IMPLICATIONS Shortening of N-terminus or mutation of glycosylation sites located within the N-terminus enhanced receptor internalization but impaired the surface expression of D2 receptors. The N-terminal region of the D2 receptor, in a sequence-specific manner, controls the receptor's conformation and integration into the plasma membrane, which determine its subcellular localization, intracellular trafficking and signalling properties. PMID:22117524

Rpas and Tls Tecniques for Archaeological Survey: the Case Study of the Archaeological Site of Eraclea Minoa (italy)

NASA Astrophysics Data System (ADS)

Lo Brutto, M.; Sciortino, R.; Garraffa, A.

2017-02-01

Digital documentation and 3D modelling of archaeological sites are important for understanding, definition and recognition of the values of the sites and of the archaeological finds. The most part of archaeological sites are outdoor location, but a cover to preserve the ruins protects often parts of the sites. The possibility to acquire data with different techniques and merge them by using a single reference system allows creating multi-parties models in which 3D representations of the individual objects can be inserted. The paper presents the results of a recent study carried out by Geomatics Laboratory of University of Palermo for the digital documentation and 3D modelling of Eraclea Minoa archaeological site. This site is located near Agrigento, in the south of Sicily (Italy) and is one of the most famous ancient Greek colonies of Sicily. The paper presents the results of the integration of different data source to survey the Eraclea Minoa archaeological site. The application of two highly versatile recording systems, the TLS (Terrestrial Laser Scanning) and the RPAS (Remotely Piloted Aircraft System), allowed the Eraclea Minoa site to be documented in high resolution and with high accuracy. The integration of the two techniques has demonstrated the possibility to obtain high quality and accurate 3D models in archaeological survey.

Scaleable Clean Aluminum Melting Systems

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

Han, Q.; Das, S.K.

2008-02-15

The project entitled 'Scaleable Clean Aluminum Melting Systems' was a Cooperative Research and Development Agreements (CRADAs) between Oak Ridge National Laboratory (ORNL) and Secat Inc. The three-year project was initially funded for the first year and was then canceled due to funding cuts at the DOE headquarters. The limited funds allowed the research team to visit industrial sites and investigate the status of using immersion heaters for aluminum melting applications. Primary concepts were proposed on the design of furnaces using immersion heaters for melting. The proposed project can continue if the funding agency resumes the funds to this research. Themore » objective of this project was to develop and demonstrate integrated, retrofitable technologies for clean melting systems for aluminum in both the Metal Casting and integrated aluminum processing industries. The scope focused on immersion heating coupled with metal circulation systems that provide significant opportunity for energy savings as well as reduction of melt loss in the form of dross. The project aimed at the development and integration of technologies that would enable significant reduction in the energy consumption and environmental impacts of melting aluminum through substitution of immersion heating for the conventional radiant burner methods used in reverberatory furnaces. Specifically, the program would couple heater improvements with furnace modeling that would enable cost-effective retrofits to a range of existing furnace sizes, reducing the economic barrier to application.« less

Integrated Propulsion Data System Public Web Site

NASA Technical Reports Server (NTRS)

Hamilton, Kimberly

2001-01-01

The Integrated Propulsion Data System's (IPDS) focus is to provide technologically-advanced philosophies of doing business at SSC that will enhance the existing operations, engineering and management strategies and provide insight and metrics to assess their daily impacts, especially as related to the Propulsion Test Directorate testing scenarios for the 21st Century.

A Numerical Study for Groundwater Flow, Heat and Solute Transport Associated with Operation of Open-loop Geothermal System in Alluvial Aquifer

NASA Astrophysics Data System (ADS)

Park, D. K.; Bae, G. O.; Lee, K. K.

2014-12-01

The open-loop geothermal system directly uses a relatively stable temperature of groundwater for cooling and heating in buildings and thus has been known as an eco-friendly, energy-saving, and cost-efficient technique. The facility for this system was installed at a site located near Paldang-dam in Han-river, Korea. Because of the well-developed alluvium, the site might be appropriate to application of this system requiring extraction and injection of a large amount of groundwater. A simple numerical experiment assuming various hydrogeologic conditions demonstrated that regional groundwater flow direction was the most important factor for efficient operation of facility in this site having a highly permeable layer. However, a comparison of river stage data and groundwater level measurements showed that the daily and seasonal controls of water level at Paldang-dam have had a critical influence on the regional groundwater flow in the site. Moreover, nitrate concentrations measured in the monitoring wells gave indication of the effect of agricultural activities around the facility on the groundwater quality. The facility operation, such as extraction and injection of groundwater, will obviously affect transport of the agricultural contaminant and, maybe, it will even cause serious problems in the normal operation. Particularly, the high-permeable layer in this aquifer must be a preferential path for quick spreadings of thermal and contaminant plumes. The objective of this study was to find an efficient, safe and stable operation plan of the open-loop geothermal system installed in this site having the complicated conditions of highly permeable layer, variable regional groundwater flow, and agricultural contamination. Numerical simulations for groundwater flow, heat and solute transport were carried out to analyze all the changes in groundwater level and flow, temperature, and quality according to the operation, respectively. Results showed that an operation plan for only the thermal efficiency of system cannot be the best in aspect of safe and stable operation related to groundwater quality. All these results concluded that it is essential to understand various and site-specific conditions of the site in a more integrated approach for the successful application of the open-loop geothermal system.

Low-cost distributed solar-thermal-electric power generation

NASA Astrophysics Data System (ADS)

Der Minassians, Artin; Aschenbach, Konrad H.; Sanders, Seth R.

2004-01-01

Due to their high relative cost, solar electric energy systems have yet to be exploited on a widespread basis. It is believed in the energy community that a technology similar to photovoltaic (PV), but offered at about $1/W would lead to widespread deployment at residential and commercial sites. This paper addresses the investigation and feasibility study of a low-cost solar thermal electricity generation technology, suitable for distributed deployment. Specifically, we discuss a system based on nonimaging solar concentrators, integrated with free-piston Stirling engine devices incorporating integrated electric generation. We target concentrator-collector operation at moderate temperatures, in the range of 125°C to 150°C. This temperature is consistent with use of optical concentrators with concentration ratios on the order of 1-2. These low ratio concentrators admit wide angles of radiation acceptance and are thus compatible with no diurnal tracking, and no or only a few seasonal adjustments. Thus, costs and reliability hazards associated with tracking hardware systems are avoided. Further, we note that in the intended application, there is no shortage of incident solar energy, but rather it is the capital cost of the solar-electric system that is most precious. Thus, we outline a strategy for exploiting solar resources in a cost constrained manner. The paper outlines design issues, and a specific design for an appropriately dimensioned free-piston Stirling engine. Only standard low-cost materials and manufacturing methods are required to realize such a machine.

Microclimate landscape design at southern integrated terminal Bandar Tasik Selatan, Kuala Lumpur

NASA Astrophysics Data System (ADS)

Phin, L. H.; Krisantia, I.

2018-01-01

Bandar Tasik Selatan is the integrated transport terminal has high energy consuming, high carbon emission and poor linkage. However, microclimate can be reduced through landscape design. This paper is a study to achieve energy efficiency and improve microclimate in the urban area. The research area is at Southern integrated terminal Bandar Tasik Selatan Kuala Lumpur Malaysia. It is carried out through a case study and microclimate analyzed using System Modeling method. System modelling using in this research is system energy budget of the microclimate at a site is a balance between the radiant energy supplied and the energy removed by all consumers. The finding indicated the microclimatic components that can be modified through landscape design are solar radiation, wind and precipitation can create thermal comfort, energy efficiency and others benefits.Through this research, provide more green space to achieve energy efficiency and improve microclimate of the site, introducing vertical landscape and proper planting selection to improve air quality, introducing green energy as part of the source of power supply and to promote integration of terminal building and rail systems by unify them using softscape

Siting Issues for Solar Thermal Power Plants with Small Community Applications

NASA Technical Reports Server (NTRS)

Holbeck, J. J.; Ireland, S. J.

1978-01-01

Technologies for solar thermal plants are being developed to provide energy alternatives for the future. Implementation of these plants requires consideration of siting issues as well as power system technology. While many conventional siting considerations are applicable, there is also a set of unique siting issues for solar thermal plants. Early experimental plants will have special siting considerations. The siting issues associated with small, dispersed solar thermal power plants in the 1 to 10 MWe power range for utility/small community applications are considered. Some specific requirements refer to the first 1 MWe engineering experiment for the Small Power Systems Applications (SPSA) Project. The siting issues themselves are discussed in three categories: (1) system resource requirements, (2) environmental effects on the system, and (3) potential impact of the plant on the environment. Within these categories, specific issues are discussed in a qualitative manner. Examples of limiting factors for some issues are taken from studies of other solar systems.

Polymeric drug delivery systems for intraoral site-specific chemoprevention of oral cancer.

PubMed

Desai, Kashappa Goud H

2018-04-01

Oral cancer is among the most prevalent cancers in the world. Moreover, it is one of the major health problems and causes of death in many regions of the world. The traditional treatment modalities include surgical removal, radiation therapy, systemic chemotherapy, or a combination of these methods. In recent decades, there has been significant interest in intraoral site-specific chemoprevention via local drug delivery using polymeric systems. Because of its easy accessibility and clear visibility, the oral mucosa is amenable for local drug delivery. A variety of polymeric systems-such as gels, tablets, films, patches, injectable systems (e.g., millicylindrical implants, microparticles, and in situ-forming depots), and nanosized carriers (e.g., polymeric nanoparticles, nanofibers, polymer-drug conjugates, polymeric micelles, nanoliposomes, nanoemulsions, and polymersomes)-have been developed and evaluated for the local delivery of natural and synthetic chemopreventive agents. The findings of in vitro, ex vivo, and in vivo studies and the positive outcome of clinical trials demonstrate that intraoral site-specific drug delivery is an attractive, highly effective and patient-friendly strategy for the management of oral cancer. Intraoral site-specific drug delivery provides unique therapeutic advantages when compared to systemic chemotherapy. Moreover, intraoral drug delivery systems are self-administrable and can be removed when needed, increasing patient compliance. This article covers important aspects and advances related to the design, development, and efficacy of polymeric systems for intraoral site-specific drug delivery. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1383-1413, 2018. © 2017 Wiley Periodicals, Inc.

The space station integrated refuse management system

NASA Technical Reports Server (NTRS)

Anderson, Loren A.

1988-01-01

The design and development of an Integrated Refuse Management System for the proposed International Space Station was performed. The primary goal was to make use of any existing potential energy or material properties that refuse may possess. The secondary goal was based on the complete removal or disposal of those products that could not, in any way, benefit astronauts' needs aboard the Space Station. The design of a continuous living and experimental habitat in space has spawned the need for a highly efficient and effective refuse management system capable of managing nearly forty-thousand pounds of refuse annually. To satisfy this need, the following four integrable systems were researched and developed: collection and transfer; recycle and reuse; advance disposal; and propulsion assist in disposal. The design of a Space Station subsystem capable of collecting and transporting refuse from its generation site to its disposal and/or recycling site was accomplished. Several methods of recycling or reusing refuse in the space environment were researched. The optimal solution was determined to be the method of pyrolysis. The objective of removing refuse from the Space Station environment, subsequent to recycling, was fulfilled with the design of a jettison vehicle. A number of jettison vehicle launch scenarios were analyzed. Selection of a proper disposal site and the development of a system to propel the vehicle to that site were completed. Reentry into the earth atmosphere for the purpose of refuse incineration was determined to be the most attractive solution.

Insights from native mass spectrometry approaches for top- and middle- level characterization of site-specific antibody-drug conjugates

PubMed Central

Botzanowski, Thomas; Erb, Stéphane; Hernandez-Alba, Oscar; Ehkirch, Anthony; Colas, Olivier; Wagner-Rousset, Elsa; Rabuka, David; Beck, Alain; Drake, Penelope M.; Cianférani, Sarah

2017-01-01

ABSTRACT Antibody-drug conjugates (ADCs) have emerged as a family of compounds with promise as efficient immunotherapies. First-generation ADCs were generated mostly via reactions on either lysine side-chain amines or cysteine thiol groups after reduction of the interchain disulfide bonds, resulting in heterogeneous populations with a variable number of drug loads per antibody. To control the position and the number of drug loads, new conjugation strategies aiming at the generation of more homogeneous site-specific conjugates have been developed. We report here the first multi-level characterization of a site-specific ADC by state-of-the-art mass spectrometry (MS) methods, including native MS and its hyphenation to ion mobility (IM-MS). We demonstrate the versatility of native MS methodologies for site-specific ADC analysis, with the unique ability to provide several critical quality attributes within one single run, along with a direct snapshot of ADC homogeneity/heterogeneity without extensive data interpretation. The capabilities of native IM-MS to directly access site-specific ADC conformational information are also highlighted. Finally, the potential of these techniques for assessing an ADC's heterogeneity/homogeneity is illustrated by comparing the analytical characterization of a site-specific DAR4 ADC to that of first-generation ADCs. Altogether, our results highlight the compatibility, versatility, and benefits of native MS approaches for the analytical characterization of all types of ADCs, including site-specific conjugates. Thus, we envision integrating native MS and IM-MS approaches, even in their latest state-of-the-art forms, into workflows that benchmark bioconjugation strategies. PMID:28406343

An open-source and low-cost monitoring system for precision enology.

PubMed

Di Gennaro, Salvatore Filippo; Matese, Alessandro; Mancin, Mirko; Primicerio, Jacopo; Palliotti, Alberto

2014-12-05

Winemaking is a dynamic process, where microbiological and chemical effects may strongly differentiate products from the same vineyard and even between wine vats. This high variability means an increase in work in terms of control and process management. The winemaking process therefore requires a site-specific approach in order to optimize cellar practices and quality management, suggesting a new concept of winemaking, identified as Precision Enology. The Institute of Biometeorology of the Italian National Research Council has developed a wireless monitoring system, consisting of a series of nodes integrated in barrel bungs with sensors for the measurement of wine physical and chemical parameters in the barrel. This paper describes an open-source evolution of the preliminary prototype, using Arduino-based technology. Results have shown good performance in terms of data transmission and accuracy, minimal size and power consumption. The system has been designed to create a low-cost product, which allows a remote and real-time control of wine evolution in each barrel, minimizing costs and time for sampling and laboratory analysis. The possibility of integrating any kind of sensors makes the system a flexible tool that can satisfy various monitoring needs.

A patient centered electronic health: eHealth system development.

PubMed

Schiza, Eirini C; Neokleous, Kleanthis C; Petkov, Nikolai; Schizas, Christos N

2015-01-01

Medical practice and patient-doctor relationship will continue improving while technology is integrated in our everyday life. In recent years the term eHealth landmarked a new era with improved health provider's skills and knowledge, and increased patient participation in medical care activities. To show why the design and implementation of a healthcare system needs to follow a specific philosophy dictated by the level of eHealth maturity of a country and its citizens. Based on the maturity level, an adaptable framework for implementing an Electronic Health System at national level is derived, guided by the Patient Centered Philosophy as defined and introduced by the EU directives. Implementation prerequisites are analyzed together with guiding principles for identifying the maturity level of an organization or country. Cyprus being a small EU country, it can be used as pilot site for the whole Europe, was chosen for this study and its maturity level analysis is presented. Recommendations that determine general steps needed to prepare the ground for an adequate patient-centered national healthcare system are accompanied. The implementation of an integrated Electronic Health Record at National level, as a prerequisite for a patient-centered eHealth environment is evidently demonstrated.

Traffic at the tmRNA Gene

PubMed Central

Williams, Kelly P.

2003-01-01

A partial screen for genetic elements integrated into completely sequenced bacterial genomes shows more significant bias in specificity for the tmRNA gene (ssrA) than for any type of tRNA gene. Horizontal gene transfer, a major avenue of bacterial evolution, was assessed by focusing on elements using this single attachment locus. Diverse elements use ssrA; among enterobacteria alone, at least four different integrase subfamilies have independently evolved specificity for ssrA, and almost every strain analyzed presents a unique set of integrated elements. Even elements using essentially the same integrase can be very diverse, as is a group with an ssrA-specific integrase of the P4 subfamily. This same integrase appears to promote damage routinely at attachment sites, which may be adaptive. Elements in arrays can recombine; one such event mediated by invertible DNA segments within neighboring elements likely explains the monophasic nature of Salmonella enterica serovar Typhi. One of a limited set of conserved sequences occurs at the attachment site of each enterobacterial element, apparently serving as a transcriptional terminator for ssrA. Elements were usually found integrated into tRNA-like sequence at the 3′ end of ssrA, at subsites corresponding to those used in tRNA genes; an exception was found at the non-tRNA-like 3′ end produced by ssrA gene permutation in cyanobacteria, suggesting that, during the evolution of new site specificity by integrases, tropism toward a conserved 3′ end of an RNA gene may be as strong as toward a tRNA-like sequence. The proximity of ssrA and smpB, which act in concert, was also surveyed. PMID:12533482

Knowledge-based approach to system integration

NASA Technical Reports Server (NTRS)

Blokland, W.; Krishnamurthy, C.; Biegl, C.; Sztipanovits, J.

1988-01-01

To solve complex problems one can often use the decomposition principle. However, a problem is seldom decomposable into completely independent subproblems. System integration deals with problem of resolving the interdependencies and the integration of the subsolutions. A natural method of decomposition is the hierarchical one. High-level specifications are broken down into lower level specifications until they can be transformed into solutions relatively easily. By automating the hierarchical decomposition and solution generation an integrated system is obtained in which the declaration of high level specifications is enough to solve the problem. We offer a knowledge-based approach to integrate the development and building of control systems. The process modeling is supported by using graphic editors. The user selects and connects icons that represent subprocesses and might refer to prewritten programs. The graphical editor assists the user in selecting parameters for each subprocess and allows the testing of a specific configuration. Next, from the definitions created by the graphical editor, the actual control program is built. Fault-diagnosis routines are generated automatically as well. Since the user is not required to write program code and knowledge about the process is present in the development system, the user is not required to have expertise in many fields.

Complex genomic rearrangement in CCS-LacZ transgenic mice.

PubMed

Stroud, Dina Myers; Darrow, Bruce J; Kim, Sang Do; Zhang, Jie; Jongbloed, Monique R M; Rentschler, Stacey; Moskowitz, Ivan P G; Seidman, Jonathan; Fishman, Glenn I

2007-02-01

The cardiac conduction system (CCS)-lacZ insertional mouse mutant strain genetically labels the developing and mature CCS. This pattern of expression is presumed to reflect the site of transgene integration rather than regulatory elements within the transgene proper. We sought to characterize the genomic structure of the integration locus and identify nearby gene(s) that might potentially confer the observed CCS-specific transcription. We found rearrangement of chromosome 7 between regions D1 and E1 with altered transcription of multiple genes in the D1 region. Several lines of evidence suggested that regulatory elements from at least one gene, Slco3A1, influenced CCS-restricted reporter gene expression. In embryonic hearts, Slco3A1 was expressed in a spatial pattern similar to the CCS-lacZ transgene and was similarly neuregulin-responsive. At later stages, however, expression patterns of the transgene and Slco3A1 diverged, suggesting that the Slco3A1 locus may be necessary, but not sufficient to confer CCS-specific transgene expression in the CCS-lacZ line. (c) 2007 Wiley-Liss, Inc.

Role of phytohormones in insect-specific plant reactions

PubMed Central

Erb, Matthias; Meldau, Stefan; Howe, Gregg A.

2012-01-01

The capacity to perceive and respond is integral to biological immune systems, but to what extent can plants specifically recognize and respond to insects? Recent findings suggest that plants possess surveillance systems that are able to detect general patterns of cellular damage as well as highly specific herbivore-associated cues. The jasmonate (JA) pathway has emerged as the major signaling cassette that integrates information perceived at the plant–insect interface into broad-spectrum defense responses. Specificity can be achieved via JA-independent processes and spatio-temporal changes of JA-modulating hormones, including ethylene, salicylic acid, abscisic acid, auxin, cytokinins, brassinosteroids and gibberellins. The identification of receptors and ligands and an integrative view of hormone-mediated response systems are crucial to understand specificity in plant immunity to herbivores. PMID:22305233

Proteome-wide Light/Dark Modulation of Thiol Oxidation in Cyanobacteria Revealed by Quantitative Site-specific Redox Proteomics*

PubMed Central

Guo, Jia; Nguyen, Amelia Y.; Dai, Ziyu; Su, Dian; Gaffrey, Matthew J.; Moore, Ronald J.; Jacobs, Jon M.; Monroe, Matthew E.; Smith, Richard D.; Koppenaal, David W.; Pakrasi, Himadri B.; Qian, Wei-Jun

2014-01-01

Reversible protein thiol oxidation is an essential regulatory mechanism of photosynthesis, metabolism, and gene expression in photosynthetic organisms. Herein, we present proteome-wide quantitative and site-specific profiling of in vivo thiol oxidation modulated by light/dark in the cyanobacterium Synechocystis sp. PCC 6803, an oxygenic photosynthetic prokaryote, using a resin-assisted thiol enrichment approach. Our proteomic approach integrates resin-assisted enrichment with isobaric tandem mass tag labeling to enable site-specific and quantitative measurements of reversibly oxidized thiols. The redox dynamics of ∼2,100 Cys-sites from 1,060 proteins under light, dark, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (a photosystem II inhibitor) conditions were quantified. In addition to relative quantification, the stoichiometry or percentage of oxidation (reversibly oxidized/total thiols) for ∼1,350 Cys-sites was also quantified. The overall results revealed broad changes in thiol oxidation in many key biological processes, including photosynthetic electron transport, carbon fixation, and glycolysis. Moreover, the redox sensitivity along with the stoichiometric data enabled prediction of potential functional Cys-sites for proteins of interest. The functional significance of redox-sensitive Cys-sites in NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, peroxiredoxin (AhpC/TSA family protein Sll1621), and glucose 6-phosphate dehydrogenase was further confirmed with site-specific mutagenesis and biochemical studies. Together, our findings provide significant insights into the broad redox regulation of photosynthetic organisms. PMID:25118246

A scalable architecture for extracting, aligning, linking, and visualizing multi-Int data

NASA Astrophysics Data System (ADS)

Knoblock, Craig A.; Szekely, Pedro

2015-05-01

An analyst today has a tremendous amount of data available, but each of the various data sources typically exists in their own silos, so an analyst has limited ability to see an integrated view of the data and has little or no access to contextual information that could help in understanding the data. We have developed the Domain-Insight Graph (DIG) system, an innovative architecture for extracting, aligning, linking, and visualizing massive amounts of domain-specific content from unstructured sources. Under the DARPA Memex program we have already successfully applied this architecture to multiple application domains, including the enormous international problem of human trafficking, where we extracted, aligned and linked data from 50 million online Web pages. DIG builds on our Karma data integration toolkit, which makes it easy to rapidly integrate structured data from a variety of sources, including databases, spreadsheets, XML, JSON, and Web services. The ability to integrate Web services allows Karma to pull in live data from the various social media sites, such as Twitter, Instagram, and OpenStreetMaps. DIG then indexes the integrated data and provides an easy to use interface for query, visualization, and analysis.

Evaluation of components, subsystems, and networks for high rate, high frequency space communications

NASA Technical Reports Server (NTRS)

Kerczewski, Robert J.; Ivancic, William D.; Zuzek, John E.

1991-01-01

The development of new space communications technologies by NASA has included both commercial applications and space science requirements. NASA's Systems Integration, Test and Evaluation (SITE) Space Communication System Simulator is a hardware based laboratory simulator for evaluating space communications technologies at the component, subsystem, system, and network level, geared toward high frequency, high data rate systems. The SITE facility is well-suited for evaluation of the new technologies required for the Space Exploration Initiative (SEI) and advanced commercial systems. Described here are the technology developments and evaluation requirements for current and planned commercial and space science programs. Also examined are the capabilities of SITE, the past, present and planned future configurations of the SITE facility, and applications of SITE to evaluation of SEI technology.

Microzonation Mapping Of The Yanbu Industrial City, Western Saudi Arabia: A Multicriteria Decision Analysis Approach

NASA Astrophysics Data System (ADS)

Moustafa, Sayed, Sr.; Alarifi, Nassir S.; Lashin, Aref A.

2016-04-01

Urban areas along the western coast of Saudi Arabia are susceptible to natural disasters and environmental damages due to lack of planning. To produce a site-specific microzonation map of the rapidly growing Yanbu industrial city, spatial distribution of different hazard entities are assessed using the Analytical Hierarchal Process (AHP) together with Geographical Information System (GIS). For this purpose six hazard parameter layers are considered, namely; fundamental frequency, site amplification, soil strength in terms of effective shear-wave velocity, overburden sediment thickness, seismic vulnerability index and peak ground acceleration. The weight and rank values are determined during AHP and are assigned to each layer and its corresponding classes, respectively. An integrated seismic microzonation map was derived using GIS platform. Based on the derived map, the study area is classified into five hazard categories: very low, low, moderate high, and very high. The western and central parts of the study area, as indicated from the derived microzonation map, are categorized as a high hazard zone as compared to other surrounding places. The produced microzonation map of the current study is envisaged as a first-level assessment of the site specific hazards in the Yanbu city area, which can be used as a platform by different stakeholders in any future land-use planning and environmental hazard management.

Retrotransposon profiling of RNA polymerase III initiation sites.

PubMed

Qi, Xiaojie; Daily, Kenneth; Nguyen, Kim; Wang, Haoyi; Mayhew, David; Rigor, Paul; Forouzan, Sholeh; Johnston, Mark; Mitra, Robi David; Baldi, Pierre; Sandmeyer, Suzanne

2012-04-01

Although retroviruses are relatively promiscuous in choice of integration sites, retrotransposons can display marked integration specificity. In yeast and slime mold, some retrotransposons are associated with tRNA genes (tDNAs). In the Saccharomyces cerevisiae genome, the long terminal repeat retrotransposon Ty3 is found at RNA polymerase III (Pol III) transcription start sites of tDNAs. Ty1, 2, and 4 elements also cluster in the upstream regions of these genes. To determine the extent to which other Pol III-transcribed genes serve as genomic targets for Ty3, a set of 10,000 Ty3 genomic retrotranspositions were mapped using high-throughput DNA sequencing. Integrations occurred at all known tDNAs, two tDNA relics (iYGR033c and ZOD1), and six non-tDNA, Pol III-transcribed types of genes (RDN5, SNR6, SNR52, RPR1, RNA170, and SCR1). Previous work in vitro demonstrated that the Pol III transcription factor (TF) IIIB is important for Ty3 targeting. However, seven loci that bind the TFIIIB loader, TFIIIC, were not targeted, underscoring the unexplained absence of TFIIIB at those sites. Ty3 integrations also occurred in two open reading frames not previously associated with Pol III transcription, suggesting the existence of a small number of additional sites in the yeast genome that interact with Pol III transcription complexes.

New thinking for the boiler room.

PubMed

Rose, Wayne

2008-09-01

Wayne Rose, marketing manager at integrated plant room manufacturer Armstrong Integrated Systems, explains how increasing use of off-site manufacture, the latest 3D modelling technology, and advances in control technology, are revolutionising boiler room design and construction.

Chromatin Landscapes of Retroviral and Transposon Integration Profiles

PubMed Central

Badhai, Jitendra; Rust, Alistair G.; Rad, Roland; Hilkens, John; Berns, Anton; van Lohuizen, Maarten; Wessels, Lodewyk F. A.; de Ridder, Jeroen

2014-01-01

The ability of retroviruses and transposons to insert their genetic material into host DNA makes them widely used tools in molecular biology, cancer research and gene therapy. However, these systems have biases that may strongly affect research outcomes. To address this issue, we generated very large datasets consisting of to unselected integrations in the mouse genome for the Sleeping Beauty (SB) and piggyBac (PB) transposons, and the Mouse Mammary Tumor Virus (MMTV). We analyzed (epi)genomic features to generate bias maps at both local and genome-wide scales. MMTV showed a remarkably uniform distribution of integrations across the genome. More distinct preferences were observed for the two transposons, with PB showing remarkable resemblance to bias profiles of the Murine Leukemia Virus. Furthermore, we present a model where target site selection is directed at multiple scales. At a large scale, target site selection is similar across systems, and defined by domain-oriented features, namely expression of proximal genes, proximity to CpG islands and to genic features, chromatin compaction and replication timing. Notable differences between the systems are mainly observed at smaller scales, and are directed by a diverse range of features. To study the effect of these biases on integration sites occupied under selective pressure, we turned to insertional mutagenesis (IM) screens. In IM screens, putative cancer genes are identified by finding frequently targeted genomic regions, or Common Integration Sites (CISs). Within three recently completed IM screens, we identified 7%–33% putative false positive CISs, which are likely not the result of the oncogenic selection process. Moreover, results indicate that PB, compared to SB, is more suited to tag oncogenes. PMID:24721906

Non-invasive investigative techniques for the diachronic study of territorial compartments: a case study for the documentation and analysis of architectural complexes.

NASA Astrophysics Data System (ADS)

Di Lieto, Marco; Marchetta, Isabella; Ciriello, Rosanna; De Martino, Gregory; Della Mora, Dario

2014-05-01

The trend in the study of areas of land in their integrity and as dynamic, anthropic units in diachronic history has initiated long survey campaigns over several decades that have covered large areas mapping the evidence and attempting a reconstruction of the evolution of ancient settlements. The need for further study to disentangle the knots of modes and types of settlement boosted further investigations of targeted excavations, based on the quality and density of the findings from the field. Currently archaeological research can rely on non-invasive integrated methods to better define the areas to be investigated systematically obtaining new typologies of information and better management of time and research costs. In this paper we present a specific case study in which a variety of integrated survey methods have contributed to the documentation and analysis of monumental complexes linked to specific local contexts. The area under investigation lies in Italy, in the province of Potenza and, specifically in the town of Forenza. The survey activities, involving the entire municipality, have been running on and off for about 2 years and have already resulted in the collection of a lot of interesting data that will be useful to essential fieldwork. In particular, we carried out different types of investigation in three different sample sites: 1. monumental complex of Santa Maria de 'Armenis: to complement previous excavations which involved only a portion of the estimated area of interest, we carried out magnetometric and geo-electrical surveys aimed at a more precise definition of the true extent and interpretation of the monument in antiquity; 2. site of Monte Caruso: we carried out remote sensing using a remote-controlled UAV hexakopter drone with stereoscopic photogrammetric survey techniques aimed at the detailed documentation of the monumental evidence of a structure visible in elevation but in a context difficult to approach with traditional surveying systems; 3. wine presses in the "palmienti" district: we also tested here using remote sensing with a remote-controlled UAV hexakopter drone, introducing the automatic flight mode on waypoints in order to obtain precise photogrammetric strips and the creation of a detailed 3D model to document the complex geomorphology of the site, optimising time and the limited resources available. In conclusion, the integrated use of these non-invasive investigative techniques had the advantage of obtaining valuable information and optimising the limited resources, creating an effective working basis for future project development, and immediately usable data for informational purposes, for the sponsors.

Integrated chemical/biochemical sample collection, pre-concentration, and analysis on a digital microfluidic lab-on-a-chip platform

NASA Astrophysics Data System (ADS)

Fair, Richard B.; Khlystov, A.; Srinivasan, Vijay; Pamula, Vamsee K.; Weaver, Kathryn N.

2004-12-01

An ideal on-site chemical/biochemical analysis system must be inexpensive, sensitive, fully automated and integrated, reliable, and compatible with a broad range of samples. The advent of digital microfluidic lab-on-a-chip (LoC) technology offers such a detection system due to the advantages in portability, reduction of the volumes of the sample and reagents, faster analysis times, increased automation, low power consumption, compatibility with mass manufacturing, and high throughput. We describe progress towards integrating sample collection onto a digital microfluidic LoC that is a component of a cascade impactor device. The sample collection is performed by impacting airborne particles directly onto the surface of the chip. After the collection phase, the surface of the chip is washed with a micro-droplet of solvent. The droplet will be digitally directed across the impaction surface, dissolving sample constituents. Because of the very small droplet volume used for extraction of the sample from a wide colection area, the resulting solution is realatively concentrated and the analytes can be detected after a very short sampling time (1 min) due to such pre-concentration. After the washing phase, the droplet is mixed with specific reagents that produce colored reaction products. The concentration of the analyte is quantitatively determined by measuring absorption at target wavelengths using a simple light emitting diode and photodiode setup. Specific applications include automatic measurements of major inorganic ions in aerosols, such as sulfate, nitrate and ammonium, with a time resolution of 1 min and a detection limit of 30 nm/m3. We have already demonstrated the detection and quantification of nitroaromatic explosives without integrating the sample collection. Other applications being developed include airborne bioagent detection.

College Student Samples Are Not Always Equivalent: The Magnitude of Personality Differences Across Colleges and Universities.

PubMed

Corker, Katherine S; Donnellan, M Brent; Kim, Su Yeong; Schwartz, Seth J; Zamboanga, Byron L

2017-04-01

This research examined the magnitude of personality differences across different colleges and universities to understand (a) how much students at different colleges vary from one another and (b) whether there are site-level variables that can explain observed differences. Nearly 8,600 students at 30 colleges and universities completed a Big Five personality trait measure. Site-level information was obtained from the Integrated Postsecondary Education System database (U.S. Department of Education). Multilevel models revealed that each of the Big Five traits showed significant between-site variability, even after accounting for individual-level demographic differences. Some site-level variables (e.g., enrollment size, requiring letters of recommendation) explained between-site differences in traits, but many tests were not statistically significant. Student samples at different universities differed in terms of average levels of Big Five personality domains. This raises the possibility that personality differences may explain differences in research results obtained when studying students at different colleges and universities. Furthermore, results suggest that research that compares findings for only a few sites (e.g., much cross-cultural research) runs the risk of overgeneralizing differences between specific samples to broader group differences. These results underscore the value of multisite collaborative research efforts to enhance psychological research. © 2015 Wiley Periodicals, Inc.

Approaching a universal scaling relationship between fracture stiffness and fluid flow

NASA Astrophysics Data System (ADS)

Pyrak-Nolte, Laura J.; Nolte, David D.

2016-02-01

A goal of subsurface geophysical monitoring is the detection and characterization of fracture alterations that affect the hydraulic integrity of a site. Achievement of this goal requires a link between the mechanical and hydraulic properties of a fracture. Here we present a scaling relationship between fluid flow and fracture-specific stiffness that approaches universality. Fracture-specific stiffness is a mechanical property dependent on fracture geometry that can be monitored remotely using seismic techniques. A Monte Carlo numerical approach demonstrates that a scaling relationship exists between flow and stiffness for fractures with strongly correlated aperture distributions, and continues to hold for fractures deformed by applied stress and by chemical erosion as well. This new scaling relationship provides a foundation for simulating changes in fracture behaviour as a function of stress or depth in the Earth and will aid risk assessment of the hydraulic integrity of subsurface sites.

A telemedicine health care delivery system

NASA Technical Reports Server (NTRS)

Sanders, Jay H.

1991-01-01

The Interactive Telemedicine Systems (ITS) system was specifically developed to address the ever widening gap between our medical care expertise and our medical care delivery system. The frustrating reality is that as our knowledge of how to diagnose and treat medical conditions has continued to advance, the system to deliver that care has remained in an embryonic stage. This has resulted in millions of people being denied their most basic health care needs. Telemedicine utilizes an interactive video system integrated with biomedical telemetry that allows a physician at a base station specialty medical complex or teaching hospital to examine and treat a patient at multiple satellite locations, such as rural hospitals, ambulatory health centers, correctional institutions, facilities caring for the elderly, community hospital emergency departments, or international health facilities. Based on the interactive nature of the system design, the consulting physician at the base station can do a complete history and physical examination, as if the patient at the satellite site was sitting in the physician's office. This system is described.

3D geophysical imaging for site-specific characterization plan of an old landfill.

PubMed

Di Maio, R; Fais, S; Ligas, P; Piegari, E; Raga, R; Cossu, R

2018-06-01

As it is well-known, the characterization plan of an old landfill site is the first stage of the project for the treatment and reclamation of contaminated lands. It is a preliminary in-situ study, with collection of data related to pollution phenomena, and is aimed at defining the physical properties and the geometry of fill materials as well as the possible migration paths of pollutants to the surrounding environmental targets (subsoil and groundwater). To properly evaluate the extent and potential for subsoil contamination, waste volume and possible leachate emissions from the landfill have to be assessed. In such perspective, the integrated use of geophysical methods is an important tool as it allows a detailed 3D representation of the whole system, i.e. waste body and hosting environment (surrounding rocks). This paper presents a very accurate physical and structural characterization of an old landfill and encasing rocks obtained by an integrated analysis of data coming from a multi-methodological geophysical exploration. Moreover, drillings were carried out for waste sampling and characterization of the landfill body, as well as for calibration of the geophysical modeling. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effectiveness of an Integrated, Trauma-Informed Approach to Treating Women with Co-Occurring Disorders and Histories of Trauma: The Los Angeles Site Experience

ERIC Educational Resources Information Center

Gatz, Margaret; Brown, Vivian; Hennigan, Karen; Rechberger, Elke; O'Keefe, Maura; Rose, Tara; Bjelajac, Paula

2007-01-01

Effectiveness of an integrated trauma-informed approach to treating women with co-occurring disorders and histories of trauma was evaluated. Baseline and 12-month assessments were completed by 136 intervention- and 177 comparison-group women. The intervention group received Seeking Safety, a trauma-specific group treatment focusing on safety and…

[The role of integrons in dissemination of antibiotic resistance].

PubMed

Ploy, M C; Lambert, T; Gassama, A; Denis, F

2000-01-01

Bacteria can transfer genetic information to get protection against most antibiotics. The acquisition of resistance genes involves genetic mobile elements such as plasmids and transposons. Another genetic structures, named integrons, have been described and contain one or more gene cassettes located at a specific site. Integrons contain an intI gene encoding a site-specific recombinase belonging to the integrase family and a recombination site attI. A gene cassette includes an open reading frame and, at the 3'-end, a recombination site attC. Integration or excision of cassettes occurs by a site-specific recombination mechanism catalyzed by the integrase. However, insertion can rarely occur, at non-specific sites leading to a stable situation for the cassette. Cassettes are transcribed from a common promoter located in the 5'-conserved segment and expression of distal genes is reduced by the presence of upstream cassettes. Most gene cassettes encode antibiotic resistant determinants but antiseptic resistant genes have also been described. Integrons seem to have a major role in the spread of multidrug resistance in Gram-negative bacteria but integrons in Gram-positive bacteria have been recently described. Moreover, the finding of super-integrons with gene cassettes coding for other determinants (biochemical functions, virulence factors) in different Gram negative bacteria suggests that integrons are probably implied in bacterial genome evolution.

Word-to-text integration: ERP evidence for semantic and orthographic effects in Chinese.

PubMed

Chen, Lin; Fang, Xiaoping; Perfetti, Charles A

2017-05-01

Although writing systems affect reading at the level of word identification, one expects writing system to have minimal effects on comprehension processes. We tested this assumption by recording ERPs while native Chinese speakers read short texts for comprehension in the word-to-text integration (WTI) paradigm to compare with studies of English using this paradigm. Of interest was the ERP on a 2-character word that began the second sentence of the text, with the first sentence varied to manipulate co-reference with the critical word in the second sentence. A paraphrase condition in which the critical word meaning was coreferential with a word in the first sentence showed a reduced N400 reduction. Consistent with results in English, this N400 effect suggests immediate integration of a Chinese 2-character word with the meaning of the text. Chinese allows an additional test of a morpheme effect when one character of a two-character word is repeated across the sentence boundary, thus having both orthographic and meaning overlap. This shared morpheme condition showed no effect during the timeframe when orthographic effects are observed (e.g. N200), nor did it show an N400 effect. However, character repetition did produce an N400 reduction on parietal sites regardless it represented the same morpheme or a different one. The results indicate that the WTI integration effect is general across writing systems at the meaning level, but that the orthographic form nonetheless has an effect, and is specifically functional in Chinese reading.

Autonomous Integrated Receive System (AIRS) requirements definition. Volume 4: Functional specification for the prototype Automated Integrated Receive System (AIRS)

NASA Technical Reports Server (NTRS)

Chie, C. M.

1984-01-01

The functional requirements for the performance, design, and testing for the prototype Automated Integrated Receive System (AIRS) to be demonstrated for the TDRSS S-Band Single Access Return Link are presented.

Photovoltaic balance-of-system designs and costs at PVUSA

NASA Astrophysics Data System (ADS)

Reyes, A. B.; Jennings, C.

1995-05-01

This report is one in a series of 1994-1995 PVUSA reports that document PVUSA lessons learned at demonstration sites in California and Texas. During the last 7 years (1988 to 1994), 16 PV systems ranging from 20 kW to 500 kW have been installed. Six 20-kW emerging module technology (EMT) arrays and three turnkey (i.e., vendor designed and integrated) utility-scale systems were procured and installed at PVUSA's main test site in Davis, California. PVUSA host utilities have installed a total of seven EMT arrays and utility-scale systems in their service areas. Additional systems at Davis and host utility sites are planned. One of PVUSA's key objectives is to evaluate the performance, reliability, and cost of PV balance-of-system (BOS). In the procurement stage PVUSA encouraged innovative design to improve upon present practice by reducing maintenance, improving reliability, or lowering manufacturing or construction costs. The project team worked closely with suppliers during the design stage not only to ensure designs met functional and safety specifications, but to provide suggestions for improvement. This report, intended for the photovoltaic (PV) industry and for utility project managers and engineers considering PV plant construction and ownership, documents PVUSA utility-scale system design and cost lessons learned. Complementary PVUSA topical reports document: construction and safety experience; five-year assessment of EMTs; validation of the Kerman 500-kW grid-support PV plant benefits; PVUSA instrumentation and data analysis techniques; procurement, acceptance, and rating practices for PV power plants; experience with power conditioning units and power quality.

Idaho National Laboratory Site Pollution Prevention Plan

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

E. D. Sellers

2007-03-01

It is the policy of the Department of Energy (DOE) that pollution prevention and sustainable environmental stewardship will be integrated into DOE operations as a good business practice to reduce environmental hazards, protect environmental resources, avoid pollution control costs, and improve operational efficiency and mission sustainability. In furtherance of this policy, DOE established five strategic, performance-based Pollution Prevention (P2) and Sustainable Environmental Stewardship goals and included them as an attachment to DOE O 450.1, Environmental Protection Program. These goals and accompanying strategies are to be implemented by DOE sites through the integration of Pollution Prevention into each site's Environmental Managementmore » System (EMS). This document presents a P2 and Sustainability Program and corresponding plan pursuant to DOE Order 450.1 and DOE O 435.1, Radioactive Waste Management. This plan is also required by the state of Idaho, pursuant to the Resource Conservation and Recovery Act (RCRA) partial permit. The objective of this document is to describe the Idaho National Laboratory (INL) Site P2 and Sustainability Program. The purpose of the program is to decrease the environmental footprint of the INL Site while providing enhanced support of its mission. The success of the program is dependent on financial and management support. The signatures on the previous page indicate INL, ICP, and AMWTP Contractor management support and dedication to the program. P2 requirements have been integrated into working procedures to ensure an effective EMS as part of an Integrated Safety Management System (ISMS). This plan focuses on programmatic functions which include environmentally preferable procurement, sustainable design, P2 and Sustainability awareness, waste generation and reduction, source reduction and recycling, energy management, and pollution prevention opportunity assessments. The INL Site P2 and Sustainability Program is administratively managed by the INL Site P2 Coordinator. Development and maintenance of this overall INL Site plan is ultimately the responsibility of DOE-ID. This plan is applicable to all INL Site contractors except those at the Naval Reactors Facility.« less

A Conserved Non-Canonical Docking Mechanism Regulates the Binding of Dual Specificity Phosphatases to Cell Integrity Mitogen-Activated Protein Kinases (MAPKs) in Budding and Fission Yeasts

PubMed Central

Sacristán-Reviriego, Almudena; Madrid, Marisa; Cansado, José; Martín, Humberto; Molina, María

2014-01-01

Dual-specificity MAPK phosphatases (MKPs) are essential for the negative regulation of MAPK pathways. Similar to other MAPK-interacting proteins, most MKPs bind MAPKs through specific docking domains known as D-motifs. However, we found that the Saccharomyces cerevisiae MKP Msg5 binds the MAPK Slt2 within the cell wall integrity (CWI) pathway through a distinct motif (IYT). Here, we demonstrate that the IYT motif mediates binding of the Msg5 paralogue Sdp1 to Slt2 as well as of the MKP Pmp1 to its CWI MAPK counterpart Pmk1 in the evolutionarily distant yeast Schizosaccharomyces pombe. As a consequence, removal of the IYT site in Msg5, Sdp1 and Pmp1 reduces MAPK trapping caused by the overexpression of catalytically inactive versions of these phosphatases. Accordingly, an intact IYT site is necessary for inactive Sdp1 to prevent nuclear accumulation of Slt2. We also show that both Ile and Tyr but not Thr are essential for the functionality of the IYT motif. These results provide mechanistic insight into MKP-MAPK interplay and stress the relevance of this conserved non-canonical docking site in the regulation of the CWI pathway in fungi. PMID:24465549

Efficient Genome Editing in Induced Pluripotent Stem Cells with Engineered Nucleases In Vitro.

PubMed

Termglinchan, Vittavat; Seeger, Timon; Chen, Caressa; Wu, Joseph C; Karakikes, Ioannis

2017-01-01

Precision genome engineering is rapidly advancing the application of the induced pluripotent stem cells (iPSCs) technology for in vitro disease modeling of cardiovascular diseases. Targeted genome editing using engineered nucleases is a powerful tool that allows for reverse genetics, genome engineering, and targeted transgene integration experiments to be performed in a precise and predictable manner. However, nuclease-mediated homologous recombination is an inefficient process. Herein, we describe the development of an optimized method combining site-specific nucleases and the piggyBac transposon system for "seamless" genome editing in pluripotent stem cells with high efficiency and fidelity in vitro.

Aquifer response to stream-stage and recharge variations. II. Convolution method and applications

USGS Publications Warehouse

Barlow, P.M.; DeSimone, L.A.; Moench, A.F.

2000-01-01

In this second of two papers, analytical step-response functions, developed in the companion paper for several cases of transient hydraulic interaction between a fully penetrating stream and a confined, leaky, or water-table aquifer, are used in the convolution integral to calculate aquifer heads, streambank seepage rates, and bank storage that occur in response to streamstage fluctuations and basinwide recharge or evapotranspiration. Two computer programs developed on the basis of these step-response functions and the convolution integral are applied to the analysis of hydraulic interaction of two alluvial stream-aquifer systems in the northeastern and central United States. These applications demonstrate the utility of the analytical functions and computer programs for estimating aquifer and streambank hydraulic properties, recharge rates, streambank seepage rates, and bank storage. Analysis of the water-table aquifer adjacent to the Blackstone River in Massachusetts suggests that the very shallow depth of water table and associated thin unsaturated zone at the site cause the aquifer to behave like a confined aquifer (negligible specific yield). This finding is consistent with previous studies that have shown that the effective specific yield of an unconfined aquifer approaches zero when the capillary fringe, where sediment pores are saturated by tension, extends to land surface. Under this condition, the aquifer's response is determined by elastic storage only. Estimates of horizontal and vertical hydraulic conductivity, specific yield, specific storage, and recharge for a water-table aquifer adjacent to the Cedar River in eastern Iowa, determined by the use of analytical methods, are in close agreement with those estimated by use of a more complex, multilayer numerical model of the aquifer. Streambank leakance of the semipervious streambank materials also was estimated for the site. The streambank-leakance parameter may be considered to be a general (or lumped) parameter that accounts not only for the resistance of flow at the river-aquifer boundary, but also for the effects of partial penetration of the river and other near-stream flow phenomena not included in the theoretical development of the step-response functions.Analytical step-response functions, developed for several cases of transient hydraulic interaction between a fully penetrating stream and a confined, leaky, or water-table aquifer, are used in the convolution integral to calculate aquifer heads, streambank seepage rates, and bank storage that occur in response to stream-stage fluctuations and basinwide recharge or evapotranspiration. Two computer programs developed on the basis of these step-response functions and the convolution integral are applied to the analysis of hydraulic interaction of two alluvial stream-aquifer systems. These applications demonstrate the utility of the analytical functions and computer programs for estimating aquifer and streambank seepage rates and bank storage.

Implementation of Malaria Dynamic Models in Municipality Level Early Warning Systems in Colombia. Part I: Description of Study Sites

PubMed Central

Ruiz, Daniel; Cerón, Viviana; Molina, Adriana M.; Quiñónes, Martha L.; Jiménez, Mónica M.; Ahumada, Martha; Gutiérrez, Patricia; Osorio, Salua; Mantilla, Gilma; Connor, Stephen J.; Thomson, Madeleine C.

2014-01-01

As part of the Integrated National Adaptation Pilot project and the Integrated Surveillance and Control System, the Colombian National Institute of Health is working on the design and implementation of a Malaria Early Warning System framework, supported by seasonal climate forecasting capabilities, weather and environmental monitoring, and malaria statistical and dynamic models. In this report, we provide an overview of the local ecoepidemiologic settings where four malaria process-based mathematical models are currently being implemented at a municipal level. The description includes general characteristics, malaria situation (predominant type of infection, malaria-positive cases data, malaria incidence, and seasonality), entomologic conditions (primary and secondary vectors, mosquito densities, and feeding frequencies), climatic conditions (climatology and long-term trends), key drivers of epidemic outbreaks, and non-climatic factors (populations at risk, control campaigns, and socioeconomic conditions). Selected pilot sites exhibit different ecoepidemiologic settings that must be taken into account in the development of the integrated surveillance and control system. PMID:24891460

DASMI: exchanging, annotating and assessing molecular interaction data.

PubMed

Blankenburg, Hagen; Finn, Robert D; Prlić, Andreas; Jenkinson, Andrew M; Ramírez, Fidel; Emig, Dorothea; Schelhorn, Sven-Eric; Büch, Joachim; Lengauer, Thomas; Albrecht, Mario

2009-05-15

Ever increasing amounts of biological interaction data are being accumulated worldwide, but they are currently not readily accessible to the biologist at a single site. New techniques are required for retrieving, sharing and presenting data spread over the Internet. We introduce the DASMI system for the dynamic exchange, annotation and assessment of molecular interaction data. DASMI is based on the widely used Distributed Annotation System (DAS) and consists of a data exchange specification, web servers for providing the interaction data and clients for data integration and visualization. The decentralized architecture of DASMI affords the online retrieval of the most recent data from distributed sources and databases. DASMI can also be extended easily by adding new data sources and clients. We describe all DASMI components and demonstrate their use for protein and domain interactions. The DASMI tools are available at http://www.dasmi.de/ and http://ipfam.sanger.ac.uk/graph. The DAS registry and the DAS 1.53E specification is found at http://www.dasregistry.org/.

Integrated flight/propulsion control - Subsystem specifications for performance

NASA Technical Reports Server (NTRS)

Neighbors, W. K.; Rock, Stephen M.

1993-01-01

A procedure is presented for calculating multiple subsystem specifications given a number of performance requirements on the integrated system. This procedure applies to problems where the control design must be performed in a partitioned manner. It is based on a structured singular value analysis, and generates specifications as magnitude bounds on subsystem uncertainties. The performance requirements should be provided in the form of bounds on transfer functions of the integrated system. This form allows the expression of model following, command tracking, and disturbance rejection requirements. The procedure is demonstrated on a STOVL aircraft design.

Mitotic Cortical Waves Predict Future Division Sites by Encoding Positional and Size Information.

PubMed

Xiao, Shengping; Tong, Cheesan; Yang, Yang; Wu, Min

2017-11-20

Dynamic spatial patterns such as traveling waves could theoretically encode spatial information, but little is known about whether or how they are employed by biological systems, especially higher eukaryotes. Here, we show that concentric target or spiral waves of active Cdc42 and the F-BAR protein FBP17 are invoked in adherent cells at the onset of mitosis. These waves predict the future sites of cell divisions and represent the earliest known spatial cues for furrow assembly. Unlike interphase waves, the frequencies and wavelengths of the mitotic waves display size-dependent scaling properties. While the positioning role of the metaphase waves requires microtubule dynamics, spindle and microtubule-independent inhibitory signals are propagated by the mitotic waves to ensure the singularity of furrow formation. Taken together, we propose that metaphase cortical waves integrate positional and cell size information for division-plane specification in adhesion-dependent cytokinesis. Copyright © 2017 Elsevier Inc. All rights reserved.

c-Myb Binds to a Sequence in the Proximal Region of the RAG-2 Promoter and Is Essential for Promoter Activity in T-Lineage Cells

PubMed Central

Wang, Qian-Fei; Lauring, Josh; Schlissel, Mark S.

2000-01-01

The RAG-2 gene encodes a component of the V(D)J recombinase which is essential for the assembly of antigen receptor genes in B and T lymphocytes. Previously, we reported that the transcription factor BSAP (PAX-5) regulates the murine RAG-2 promoter in B-cell lines. A partially overlapping but distinct region of the proximal RAG-2 promoter was also identified as an important element for promoter activity in T cells; however, the responsible factor was unknown. In this report, we present data demonstrating that c-Myb binds to a Myb consensus site within the proximal promoter and is critical for its activity in T-lineage cells. We show that c-Myb can transactivate a RAG-2 promoter-reporter construct in cotransfection assays and that this transactivation depends on the proximal promoter Myb consensus site. By using a chromatin immunoprecipitation (ChIP) strategy, fractionation of chromatin with anti-c-Myb antibody specifically enriched endogenous RAG-2 promoter DNA sequences. DNase I genomic footprinting revealed that the c-Myb site is occupied in a tissue-specific fashion in vivo. Furthermore, an integrated RAG-2 promoter construct with mutations at the c-Myb site was not enriched in the ChIP assay, while a wild-type integrated promoter construct was enriched. Finally, this lack of binding of c-Myb to a chromosomally integrated mutant RAG-2 promoter construct in vivo was associated with a striking decrease in promoter activity. We conclude that c-Myb regulates the RAG-2 promoter in T cells by binding to this consensus c-Myb binding site. PMID:11094072

Integration of HIV/AIDS services into African primary health care: lessons learned for health system strengthening in Mozambique - a case study

PubMed Central

2010-01-01

Introduction In 2004, Mozambique, supported by large increases in international disease-specific funding, initiated a national rapid scale-up of antiretroviral treatment (ART) and HIV care through a vertical "Day Hospital" approach. Though this model showed substantial increases in people receiving treatment, it diverted scarce resources away from the primary health care (PHC) system. In 2005, the Ministry of Health (MOH) began an effort to use HIV/AIDS treatment and care resources as a means to strengthen their PHC system. The MOH worked closely with a number of NGOs to integrate HIV programs more effectively into existing public-sector PHC services. Case Description In 2005, the Ministry of Health and Health Alliance International initiated an effort in two provinces to integrate ART into the existing primary health care system through health units distributed across 23 districts. Integration included: a) placing ART services in existing units; b) retraining existing workers; c) strengthening laboratories, testing, and referral linkages; e) expanding testing in TB wards; f) integrating HIV and antenatal services; and g) improving district-level management. Discussion: By 2008, treatment was available in nearly 67 health facilities in 23 districts. Nearly 30,000 adults were on ART. Over 80,000 enrolled in the HIV/AIDS program. Loss to follow-up from antenatal and TB testing to ART services has declined from 70% to less than 10% in many integrated sites. Average time from HIV testing to ART initiation is significantly faster and adherence to ART is better in smaller peripheral clinics than in vertical day hospitals. Integration has also improved other non-HIV aspects of primary health care. Conclusion The integration approach enables the public sector PHC system to test more patients for HIV, place more patients on ART more quickly and efficiently, reduce loss-to-follow-up, and achieve greater geographic HIV care coverage compared to the vertical model. Through the integration process, HIV resources have been used to rehabilitate PHC infrastructure (including laboratories and pharmacies), strengthen supervision, fill workforce gaps, and improve patient flow between services and facilities in ways that can benefit all programs. Using aid resources to integrate and better link HIV care with existing services can strengthen wider PHC systems. PMID:20180975

LANDSAT demonstration/application and GIS integration in south central Alaska

NASA Technical Reports Server (NTRS)

Burns, A. W.; Derrenbacher, W.

1981-01-01

Automated geographic information systems were developed for two sites in Southcentral Alaska to serve as tests for both the process of integrating classified LANDSAT data into a comprehensive environmental data base and the process of using automated information in land capability/suitability analysis and environmental planning. The Big Lake test site, located approximately 20 miles north of the City of Anchorage, comprises an area of approximately 150 square miles. The Anchorage Hillside test site, lying approximately 5 miles southeast of the central part of the city, extends over an area of some 25 square miles. Map construction and content is described.

A closed-loop compressive-sensing-based neural recording system.

PubMed

Zhang, Jie; Mitra, Srinjoy; Suo, Yuanming; Cheng, Andrew; Xiong, Tao; Michon, Frederic; Welkenhuysen, Marleen; Kloosterman, Fabian; Chin, Peter S; Hsiao, Steven; Tran, Trac D; Yazicioglu, Firat; Etienne-Cummings, Ralph

2015-06-01

This paper describes a low power closed-loop compressive sensing (CS) based neural recording system. This system provides an efficient method to reduce data transmission bandwidth for implantable neural recording devices. By doing so, this technique reduces a majority of system power consumption which is dissipated at data readout interface. The design of the system is scalable and is a viable option for large scale integration of electrodes or recording sites onto a single device. The entire system consists of an application-specific integrated circuit (ASIC) with 4 recording readout channels with CS circuits, a real time off-chip CS recovery block and a recovery quality evaluation block that provides a closed feedback to adaptively adjust compression rate. Since CS performance is strongly signal dependent, the ASIC has been tested in vivo and with standard public neural databases. Implemented using efficient digital circuit, this system is able to achieve >10 times data compression on the entire neural spike band (500-6KHz) while consuming only 0.83uW (0.53 V voltage supply) additional digital power per electrode. When only the spikes are desired, the system is able to further compress the detected spikes by around 16 times. Unlike other similar systems, the characteristic spikes and inter-spike data can both be recovered which guarantes a >95% spike classification success rate. The compression circuit occupied 0.11mm(2)/electrode in a 180nm CMOS process. The complete signal processing circuit consumes <16uW/electrode. Power and area efficiency demonstrated by the system make it an ideal candidate for integration into large recording arrays containing thousands of electrode. Closed-loop recording and reconstruction performance evaluation further improves the robustness of the compression method, thus making the system more practical for long term recording.

A qualitative study of the activities performed by people involved in clinical decision support: recommended practices for success.

PubMed

Wright, Adam; Ash, Joan S; Erickson, Jessica L; Wasserman, Joe; Bunce, Arwen; Stanescu, Ana; St Hilaire, Daniel; Panzenhagen, Morgan; Gebhardt, Eric; McMullen, Carmit; Middleton, Blackford; Sittig, Dean F

2014-01-01

To describe the activities performed by people involved in clinical decision support (CDS) at leading sites. We conducted ethnographic observations at seven diverse sites with a history of excellence in CDS using the Rapid Assessment Process and analyzed the data using a series of card sorts, informed by Linstone's Multiple Perspectives Model. We identified 18 activities and grouped them into four areas. Area 1: Fostering relationships across the organization, with activities (a) training and support, (b) visibility/presence on the floor, (c) liaising between people, (d) administration and leadership, (e) project management, (f) cheerleading/buy-in/sponsorship, (g) preparing for CDS implementation. Area 2: Assembling the system with activities (a) providing technical support, (b) CDS content development, (c) purchasing products from vendors (d) knowledge management, (e) system integration. Area 3: Using CDS to achieve the organization's goals with activities (a) reporting, (b) requirements-gathering/specifications, (c) monitoring CDS, (d) linking CDS to goals, (e) managing data. Area 4: Participation in external policy and standards activities (this area consists of only a single activity). We also identified a set of recommendations associated with these 18 activities. All 18 activities we identified were performed at all sites, although the way they were organized into roles differed substantially. We consider these activities critical to the success of a CDS program. A series of activities are performed by sites strong in CDS, and sites adopting CDS should ensure they incorporate these activities into their efforts.

OTEC platform configuration and integration study. Volume I. Systems engineering and integration. Final report

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

None

1978-04-01

The economic success of an Ocean Thermal Energy Conversion (OTEC) system is highly dependent on a platform which provides adequate support for the power system, accommodates reliably the cold water pipe, and is most cost effective. The results of a study conducted for the Department of Energy to assess six generic types of platforms to determine the most satisfactory platform for severl potential sites are presented. The six platform configurations are ship, circular barge, semi-submersible, Tuned Sphere, submersible, and spar. These represent directional and symmetric types of platforms which operate on the surface, at the interface, and submerged. The fivemore » sites for this study were primarily New Orleans, Keahole Point (Hawaii), Brazil, and secondarily Key West and Puerto Rico. Electrical transmission of energy by submarine cable is the planned form of energy transmission for all sites except Brazil, where chemical conversion is to be the method of transmission. This study is devoted to the platform (or ocean systems) of the OTEC plant which is chiefly comprised of the hull and structure, the seawater system, the position control system, and miscellaneous support/assembly systems. The principal elements in the work breakdown structure for the commercial plants are presented. The assessment of the six platform configurations was conducted utilizing a baseline plan (100-MW(e) (Net)) and site (New Orleans) with variations from the baseline to cover the range of interested platforms and sites.« less

Multimedia-based decision support system for hazards recognition and abatement

DOEpatents

Czachowski, John B.; Zoldak, John T.

1998-01-01

A system for monitoring a site includes a portable data collection module used in the field to collect site specific data, and a processor module located at a central location. The data collection module displays choices of categories of findings, and then specific findings within each category. A selected specific finding is then displayed in report form with a citation to the specific code or statutory requirement, as well as a recommended course of action and an abatement date.

Comparison of Integrated Gasifier-Combined Cycle and AFB-steam turbine systems for industrial cogeneration

NASA Technical Reports Server (NTRS)

Nainiger, J. J.; Abbott, J. M.; Burns, R. K.

1981-01-01

In the cogeneration technology alternatives study (CTAS) a number of advanced coal fired systems were examined and systems using a integrated coal gasifier IGCC or a fluid bed combustor AFB were found to yield attractive cogeneration results in industrial cogeneration applications. A range of site requirements and cogeneration sizing strategies using ground rules based on CTAS were used in comparing an IGCC and an AFB. The effect of time variations in site requirements and the sensitivity to fuel and electricity price assumptions are examined. The economic alternatives of industrial or utility ownership are also considered. The results indicate that the IGCC system has potentially higher fuel and emission savings and could be an attractive option for utility ownership. The AFB steam turbine system has a potentially higher return on investment and could be attractive assuming industrial ownership.

APPLICATIONS ANALYSIS REPORT: AWD TECHNOLOGIES, INC AQUADETOX®/SVE TECHNOLOGY

EPA Science Inventory

In support of the U.S. Environmental Protection Agency’s (EPA) Superfund Innovative Technology Evaluation (SITE) Program, this report evaluates the AWD Technologies, Inc., integrated AquaDetox®/SVE treatment system for simultaneous on-site treatment of contaminated groundwater an...

Position-specific 13C distributions within propane from experiments and natural gas samples

NASA Astrophysics Data System (ADS)

Piasecki, Alison; Sessions, Alex; Lawson, Michael; Ferreira, A. A.; Santos Neto, E. V.; Ellis, Geoffrey S.; Lewan, Michael D.; Eiler, John M.

2018-01-01

Site-specific carbon isotope measurements of organic compounds potentially recover information that is lost in a conventional, 'bulk' isotopic analysis. Such measurements are useful because isotopically fractionating processes may have distinct effects at different molecular sites, and thermodynamically equilibrated populations of molecules tend to concentrate heavy isotopes in one molecular site versus another. Most recent studies of site-specific 13C in organics use specialized Nuclear Magnetic Resonance (NMR) techniques or complex chemical degradations prior to mass spectrometric measurements. Herein we present the first application of a new mass spectrometric technique that reconstructs the site-specific carbon isotope composition of propane based on measurements of the 13C/12C ratios of two or more fragment ions that sample different proportions of the terminal and central carbon sites. We apply this method to propane from laboratory experiments and natural gas samples to explore the relationships between site-specific carbon isotope composition, full-molecular δ13C, thermal maturity, and variation in organic matter precursors. Our goal is to advance the understanding of the sources and histories of short-chain alkanes within geologic systems. Our findings suggest that propane varies in its site-specific carbon isotope structure, which is correlated with increasing thermal maturity, first increasing in terminal position δ13C and then increasing in both center and terminal position δ13C. This pattern is observed in both experimental and natural samples, and is plausibly explained by a combination of site-specific, temperature-dependent isotope effects associated with conversion of different precursor molecules (kerogen, bitumen, and/or oil) to propane, differences in site-specific isotopic contents of those precursors, and possibly distillation of reactive components of those precursors with increasing maturity. We hypothesize that the largest changes in site-specific isotopic content of propane occur when bitumen and/or oil replace kerogen as the dominant precursors. If correct, this phenomenon could have significant utility for understanding gas generation in thermogenic petroleum systems.

Position-specific 13C distributions within propane from experiments and natural gas samples

USGS Publications Warehouse

Piasecki, Alison; Sessions, Alex L.; Lawson, Michael; Ferreira, A.A.; Santos Neto, E. V.; Ellis, Geoffrey S.; Lewan, Michael; Eilers, J.M.

2018-01-01

Site-specific carbon isotope measurements of organic compounds potentially recover information that is lost in a conventional, ‘bulk’ isotopic analysis. Such measurements are useful because isotopically fractionating processes may have distinct effects at different molecular sites, and thermodynamically equilibrated populations of molecules tend to concentrate heavy isotopes in one molecular site versus another. Most recent studies of site-specific 13C in organics use specialized Nuclear Magnetic Resonance (NMR) techniques or complex chemical degradations prior to mass spectrometric measurements. Herein we present the first application of a new mass spectrometric technique that reconstructs the site-specific carbon isotope composition of propane based on measurements of the 13C/12C ratios of two or more fragment ions that sample different proportions of the terminal and central carbon sites. We apply this method to propane from laboratory experiments and natural gas samples to explore the relationships between site-specific carbon isotope composition, full-molecular δ13C, thermal maturity, and variation in organic matter precursors. Our goal is to advance the understanding of the sources and histories of short-chain alkanes within geologic systems. Our findings suggest that propane varies in its site-specific carbon isotope structure, which is correlated with increasing thermal maturity, first increasing in terminal position δ13C and then increasing in both center and terminal position δ13C. This pattern is observed in both experimental and natural samples, and is plausibly explained by a combination of site-specific, temperature-dependent isotope effects associated with conversion of different precursor molecules (kerogen, bitumen, and/or oil) to propane, differences in site-specific isotopic contents of those precursors, and possibly distillation of reactive components of those precursors with increasing maturity. We hypothesize that the largest changes in site-specific isotopic content of propane occur when bitumen and/or oil replace kerogen as the dominant precursors. If correct, this phenomenon could have significant utility for understanding gas generation in thermogenic petroleum systems.

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

Mayer, J.

The U. S. Department of Energy's (DOE) Office of Environmental Management (EM) has the responsibility for cleaning up 60 sites in 22 states that were associated with the legacy of the nation's nuclear weapons program and other research and development activities. These sites are unique and many of the technologies needed to successfully disposition the associated wastes have yet to be developed or would require significant re-engineering to be adapted for future EM cleanup efforts. In 2008, the DOE-EM Engineering and Technology Program (EM-22) released the Engineering and Technology Roadmap in response to Congressional direction and the need to focusmore » on longer term activities required for the completion of the aforementioned cleanup program. One of the strategic initiatives included in the Roadmap was to enhance long term performance monitoring as defined by 'Develop and deploy cost effective long-term strategies and technologies to monitor closure sites (including soil, groundwater, and surface water) with multiple contaminants (organics, metals and radionuclides) to verify integrated long-term cleanup performance'. To support this long-term monitoring (LTM) strategic initiative, EM 22 and the Savannah River National Laboratory (SRNL) organized and held an interactive symposia, known as the 2009 DOE-EM Long-Term Monitoring Technical Forum, to define and prioritize LTM improvement strategies and products that could be realized within a 3 to 5 year investment time frame. This near-term focus on fundamental research would then be used as a foundation for development of applied programs to improve the closure and long-term performance of EM's legacy waste sites. The Technical Forum was held in Atlanta, GA on February 11-12, 2009, and attended by 57 professionals with a focus on identifying those areas of opportunity that would most effectively advance the transition of the current practices to a more effective strategy for the LTM paradigm. The meeting format encompassed three break-out sessions, which focused on needs and opportunities associated with the following LTM technical areas: (1) Performance Monitoring Tools, (2) Systems, and (3) Information Management. The specific objectives of the Technical Forum were to identify: (1) technical targets for reducing EM costs for life-cycle monitoring; (2) cost-effective approaches and tools to support the transition from active to passive remedies at EM waste sites; and (3) specific goals and objectives associated with the lifecycle monitoring initiatives outlined within the Roadmap. The first Breakout Session on LTM performance measurement tools focused on the integration and improvement of LTM performance measurement and monitoring tools that deal with parameters such as ecosystems, boundary conditions, geophysics, remote sensing, biomarkers, ecological indicators and other types of data used in LTM configurations. Although specific tools were discussed, it was recognized that the Breakout Session could not comprehensively discuss all monitoring technologies in the time provided. Attendees provided key references where other organizations have assessed monitoring tools. Three investment sectors were developed in this Breakout Session. The second Breakout Session was on LTM systems. The focus of this session was to identify new and inventive LTM systems addressing the framework for interactive parameters such as infrastructure, sensors, diagnostic features, field screening tools, state of the art characterization monitoring systems/concepts, and ecosystem approaches to site conditions and evolution. LTM systems consist of the combination of data acquisition and management efforts, data processing and analysis efforts and reporting tools. The objective of the LTM systems workgroup was to provide a vision and path towards novel and innovative LTM systems, which should be able to provide relevant, actionable information on system performance in a cost-effective manner. Two investment sectors were developed in this Breakout Session. The last Breakout Session of the Technical Forum was on LTM information management. The session focus was on the development and implementation of novel information management systems for LTM including techniques to address data issues such as: efficient management of large and diverse datasets; consistency and comparability in data management and incorporation of accurate historical information; data interpretation and information synthesis including statistical methods, modeling, and visualization; and linage of data to site management objectives and leveraging information to forge consensus among stakeholders. One investment sector was developed in this Breakout Session.« less

Determining Membrane Protein-Lipid Binding Thermodynamics Using Native Mass Spectrometry.

PubMed

Cong, Xiao; Liu, Yang; Liu, Wen; Liang, Xiaowen; Russell, David H; Laganowsky, Arthur

2016-04-06

Membrane proteins are embedded in the biological membrane where the chemically diverse lipid environment can modulate their structure and function. However, the thermodynamics governing the molecular recognition and interaction of lipids with membrane proteins is poorly understood. Here, we report a method using native mass spectrometry (MS), to determine thermodynamics of individual ligand binding events to proteins. Unlike conventional methods, native MS can resolve individual ligand binding events and, coupled with an apparatus to control the temperature, determine binding thermodynamic parameters, such as for protein-lipid interactions. We validated our approach using three soluble protein-ligand systems (maltose binding protein, lysozyme, and nitrogen regulatory protein) and obtained similar results to those using isothermal titration calorimetry and surface plasmon resonance. We also determined for the first time the thermodynamics of individual lipid binding to the ammonia channel (AmtB), an integral membrane protein from Escherichia coli. Remarkably, we observed distinct thermodynamic signatures for the binding of different lipids and entropy-enthalpy compensation for binding lipids of variable chain length. Additionally, using a mutant form of AmtB that abolishes a specific phosphatidylglycerol (PG) binding site, we observed distinct changes in the thermodynamic signatures for binding PG, implying these signatures can identify key residues involved in specific lipid binding and potentially differentiate between specific lipid binding sites.

A distance measurement between specific sites on the cytoplasmic surface of bovine rhodopsin in rod outer segment disk membranes.

PubMed

Albert, A D; Watts, A; Spooner, P; Groebner, G; Young, J; Yeagle, P L

1997-08-14

Structural information on mammalian integral membrane proteins is scarce. As part of work on an alternative approach to the structure of bovine rhodopsin, a method was devised to obtain an intramolecular distance between two specific sites on rhodopsin while in the rod outer segment disk membrane. In this report, the distance between the rhodopsin kinase phosphorylation site(s) on the carboxyl terminal and the top of the third transmembrane helix was measured on native rhodopsin. Rhodopsin was labeled with a nuclear spin label (31P) by limited phosphorylation with rhodopsin kinase. Major phosphorylation occurs at serines 343 and 338 on the carboxyl terminal. The phosphorylated rhodopsin was then specifically labeled on cysteine 140 with an electron spin label. Magic angle spinning 31P-nuclear magnetic resonance revealed the resonance arising from the phosphorylated protein. The enhancement of the transverse relaxation of this resonance by the paramagnetic spin label was observed. The strength of this perturbation was used to determine the through-space distance between the phosphorylation site(s) and the spin label position. A distance of 18 +/- 3 A was obtained.

A Real-Time Construction Safety Monitoring System for Hazardous Gas Integrating Wireless Sensor Network and Building Information Modeling Technologies.

PubMed

Cheung, Weng-Fong; Lin, Tzu-Hsuan; Lin, Yu-Cheng

2018-02-02

In recent years, many studies have focused on the application of advanced technology as a way to improve management of construction safety management. A Wireless Sensor Network (WSN), one of the key technologies in Internet of Things (IoT) development, enables objects and devices to sense and communicate environmental conditions; Building Information Modeling (BIM), a revolutionary technology in construction, integrates database and geometry into a digital model which provides a visualized way in all construction lifecycle management. This paper integrates BIM and WSN into a unique system which enables the construction site to visually monitor the safety status via a spatial, colored interface and remove any hazardous gas automatically. Many wireless sensor nodes were placed on an underground construction site and to collect hazardous gas level and environmental condition (temperature and humidity) data, and in any region where an abnormal status is detected, the BIM model will alert the region and an alarm and ventilator on site will start automatically for warning and removing the hazard. The proposed system can greatly enhance the efficiency in construction safety management and provide an important reference information in rescue tasks. Finally, a case study demonstrates the applicability of the proposed system and the practical benefits, limitations, conclusions, and suggestions are summarized for further applications.

Identification of regulatory targets of tissue-specific transcription factors: application to retina-specific gene regulation

PubMed Central

Qian, Jiang; Esumi, Noriko; Chen, Yangjian; Wang, Qingliang; Chowers, Itay; Zack, Donald J.

2005-01-01

Identification of tissue-specific gene regulatory networks can yield insights into the molecular basis of a tissue's development, function and pathology. Here, we present a computational approach designed to identify potential regulatory target genes of photoreceptor cell-specific transcription factors (TFs). The approach is based on the hypothesis that genes related to the retina in terms of expression, disease and/or function are more likely to be the targets of retina-specific TFs than other genes. A list of genes that are preferentially expressed in retina was obtained by integrating expressed sequence tag, SAGE and microarray datasets. The regulatory targets of retina-specific TFs are enriched in this set of retina-related genes. A Bayesian approach was employed to integrate information about binding site location relative to a gene's transcription start site. Our method was applied to three retina-specific TFs, CRX, NRL and NR2E3, and a number of potential targets were predicted. To experimentally assess the validity of the bioinformatic predictions, mobility shift, transient transfection and chromatin immunoprecipitation assays were performed with five predicted CRX targets, and the results were suggestive of CRX regulation in 5/5, 3/5 and 4/5 cases, respectively. Together, these experiments strongly suggest that RP1, GUCY2D, ABCA4 are novel targets of CRX. PMID:15967807

Information and image integration: project spectrum

NASA Astrophysics Data System (ADS)

Blaine, G. James; Jost, R. Gilbert; Martin, Lori; Weiss, David A.; Lehmann, Ron; Fritz, Kevin

1998-07-01

The BJC Health System (BJC) and the Washington University School of Medicine (WUSM) formed a technology alliance with industry collaborators to develop and implement an integrated, advanced clinical information system. The industry collaborators include IBM, Kodak, SBC and Motorola. The activity, called Project Spectrum, provides an integrated clinical repository for the multiple hospital facilities of the BJC. The BJC System consists of 12 acute care hospitals serving over one million patients in Missouri and Illinois. An interface engine manages transactions from each of the hospital information systems, lab systems and radiology information systems. Data is normalized to provide a consistent view for the primary care physician. Access to the clinical repository is supported by web-based server/browser technology which delivers patient data to the physician's desktop. An HL7 based messaging system coordinates the acquisition and management of radiological image data and sends image keys to the clinical data repository. Access to the clinical chart browser currently provides radiology reports, laboratory data, vital signs and transcribed medical reports. A chart metaphor provides tabs for the selection of the clinical record for review. Activation of the radiology tab facilitates a standardized view of radiology reports and provides an icon used to initiate retrieval of available radiology images. The selection of the image icon spawns an image browser plug-in and utilizes the image key from the clinical repository to access the image server for the requested image data. The Spectrum system is collecting clinical data from five hospital systems and imaging data from two hospitals. Domain specific radiology imaging systems support the acquisition and primary interpretation of radiology exams. The spectrum clinical workstations are deployed to over 200 sites utilizing local area networks and ISDN connectivity.

Study of component technologies for fuel cell on-site integrated energy systems

NASA Technical Reports Server (NTRS)

Lee, W. D.; Mathias, S.

1980-01-01

Heating, ventilation and air conditioning equipment are integrated with three types of fuel cells. System design and computer simulations are developed to utilize the thermal energy discharge of the fuel in the most cost effective manner. The fuel provides all of the electric needs and a loss of load probability analysis is used to ensure adequate power plant reliability. Equipment cost is estimated for each of the systems analyzed. A levelized annual cost reflecting owning and operating costs including the cost of money was used to select the most promising integrated system configurations. Cash flows are presented for the most promising 16 systems. Several systems for the 96 unit apartment complex (a retail store was also studied) were cost competitive with both gas and electric based conventional systems. Thermal storage is shown to be beneficial and the optimum absorption chiller sizing (waste heat recovery) in connection with electric chillers are developed. Battery storage was analyzed since the system is not electric grid connected. Advanced absorption chillers were analyzed as well. Recommendations covering financing, technical development, and policy issues are given to accelerate the commercialization of the fuel cell for on-site power generation in buildings.

Evaluation of Learning Group Approaches for Fostering Integrated Cropping Systems Management

ERIC Educational Resources Information Center

Blissett, Hana; Simmons, Steve; Jordan, Nicholas; Nelson, Kristen

2004-01-01

Cropping systems management requires integration of multiple forms of knowledge, practice, and learning by farmers, extension educators, and researchers. We evaluated the outcomes of participation in collaborative learning groups organized to address cropping systems and, specifically, challenges of integrated weed management. Groups were…

Liquid rocket booster integration study. Volume 1: Executive summary

NASA Technical Reports Server (NTRS)

1988-01-01

The impacts of introducing liquid rocket booster engines (LRB) into the Space Transportation System (STS)/Kennedy Space Center (KSC) launch environment are identified and evaluated. Proposed ground systems configurations are presented along with a launch site requirements summary. Prelaunch processing scenarios are described and the required facility modifications and new facility requirements are analyzed. Flight vehicle design recommendations to enhance launch processing are discussed. Processing approaches to integrate LRB with existing STS launch operations are evaluated. The key features and significance of launch site transition to a new STS configuration in parallel with ongoing launch activities are enumerated. This volume is the executive summary of the five volume series.

Liquid rocket booster integration study. Volume 5, part 1: Appendices

NASA Technical Reports Server (NTRS)

1988-01-01

The impacts of introducing liquid rocket booster engines (LRB) into the Space Transportation System (STS)/Kennedy Space Center (KSC) launch environment are identified and evaluated. Proposed ground systems configurations are presented along with a launch site requirements summary. Prelaunch processing scenarios are described and the required facility modifications and new facility requirements are analyzed. Flight vehicle design recommendations to enhance launch processing are discussed. Processing approaches to integrate LRB with existing STS launch operations are evaluated. The key features and significance of launch site transition to a new STS configuration in parallel with ongoing launch activities are enumerated. This volume is the appendices of the five volume series.

Liquid Rocket Booster Integration Study. Volume 2: Study synopsis

NASA Technical Reports Server (NTRS)

1988-01-01

The impacts of introducing liquid rocket booster engines (LRB) into the Space Transportation System (STS)/Kennedy Space Center (KSC) launch environment are identified and evaluated. Proposed ground systems configurations are presented along with a launch site requirements summary. Prelaunch processing scenarios are described and the required facility modifications and new facility requirements are analyzed. Flight vehicle design recommendations to enhance launch processing are discussed. Processing approaches to integrate LRB with existing STS launch operations are evaluated. The key features and significance of launch site transition to a new STS configuration in parallel with ongoing launch activities are enumerated. This volume is the study summary of the five volume series.

PROFILE: Integrated Management to Create New Breeding Habitat for Dalmatian Pelicans (Pelecanus crispus) in Greece

PubMed

Pyrovetsi

1997-09-01

/ An integrated management plan to create favorable nesting habitat for the world-endangered Dalmatian pelicans, was tested at Kerkini irrigation reservoir, a Ramsar wetland. The lake is the major wintering site of Dalmatian pelicans in Europe, where the species lives year-round without breeding. The rise of water level at the reservoir during spring (exceeding 5 m) has an impact on the whole system, including several birds, which lose their nesting habitat. Although the integrity of the wetland demands ecological restoration with changes in its hydrologic regime, local socioeconomic conditions allow only habitat level interventions. During the planning phase of the management plan, both the ecological and social context of the interventions were considered. Monitoring of all pelican habitats and populations provided the scientific basis, while a socioecological survey on knowledge/attitudes of local fishermen toward wetland identified conflicts with specific resources and planned management. To gain public support, a broad information/education program was implemented. The education program for fishermen was based on the findings of the socioecological survey. The in situ management involved experimental construction of floating rafts, platforms over water, dredged-spoil islands, and platforms at various sites of the wetland. Monitoring of the managed habitats showed that most waterbirds used them for resting and roosting. Common terns nested on the rafts, cormorants on the platforms, and Dalmatian pelicans on the man-made island. Under the prevailing hydrologic and weather conditions, islands seem to be the most suitable habitat for pelican nesting. It is concluded that wildlife habitat management should integrate the ecological component, related to the needs of the species and ecosystem, with the social one, expressed by cooperation and involvement of the local community.KEY WORDS: Integrated management; Pelican; Nesting habitat; Habitat management; Reservoir-wetland; Public participation, Greece

Use of Order Sets in Inpatient Computerized Provider Order Entry Systems: A Comparative Analysis of Usage Patterns at Seven Sites

PubMed Central

Wright, Adam; Feblowitz, Joshua C.; Pang, Justine E.; Carpenter, James D.; Krall, Michael A.; Middleton, Blackford; Sittig, Dean F.

2012-01-01

Background Many computerized provider order entry (CPOE) systems include the ability to create electronic order sets: collections of clinically-related orders grouped by purpose. Order sets promise to make CPOE systems more efficient, improve care quality and increase adherence to evidence-based guidelines. However, the development and implementation of order sets can be expensive and time-consuming and limited literature exists about their utilization. Methods Based on analysis of order set usage logs from a diverse purposive sample of seven sites with commercially- and internally-developed inpatient CPOE systems, we developed an original order set classification system. Order sets were categorized across seven non-mutually exclusive axes: admission/discharge/transfer (ADT), perioperative, condition-specific, task-specific, service-specific, convenience, and personal. In addition, 731 unique subtypes were identified within five axes: four in ADT (S=4), three in perioperative, 144 in condition-specific, 513 in task-specific, and 67 in service-specific. Results Order sets (n=1,914) were used a total of 676,142 times at the participating sites during a one-year period. ADT and perioperative order sets accounted for 27.6% and 24.2% of usage respectively. Peripartum/labor, chest pain/Acute Coronary Syndrome/Myocardial Infarction and diabetes order sets accounted for 51.6% of condition-specific usage. Insulin, angiography/angioplasty and arthroplasty order sets accounted for 19.4% of task-specific usage. Emergency/trauma, Obstetrics/Gynecology/Labor Delivery and anesthesia accounted for 32.4% of service-specific usage. Overall, the top 20% of order sets accounted for 90.1% of all usage. Additional salient patterns are identified and described. Conclusion We observed recurrent patterns in order set usage across multiple sites as well as meaningful variations between sites. Vendors and institutional developers should identify high-value order set types through concrete data analysis in order to optimize the resources devoted to development and implementation. PMID:22819199

US effort on HTS power transformers

NASA Astrophysics Data System (ADS)

Mehta, S.

2011-11-01

Waukesha Electric Systems has been working in HTS power transformers development program under the auspices of US Government Department of Energy since 1994. This presentation will describe various milestones for this program and program history along with the lessons learned along the way. Our motivations for working on this development program based on man benefits offered by HTS power transformers to power delivery systems will be discussed. Based on various issues encountered during execution of many HTS projects, DOE has set up an independent program review process that is lead by team of experts. This team reviews are integral part of all DOE HTS projects. Success of all projects would be greatly enhanced by identifying critical issues early in the program. Requiring appropriate actions to mitigate the issues before processing further will lead to proactive interrogation and incorporation of expert's ideas in the project plans. Working of this review process will be also described in this presentation. Waukesha Electric Systems team including: Superpower-Inc, Oak Ridge National Laboratory, University of Houston Center for Superconductivity and Southern California Edison company was awarded a cost share grant by US Government in 2010 for development of a fault current limiting HTS power transformer. This multi year's program will require design, manufacture, installation, and monitoring of a 28 MVA tree phase transformer installed at Irvine CA. Smart Grid demonstration site. Transformer specifications along with requirements for fault current limiting and site requirement will be discussed. Design and development of various sub systems in support of this program including: HTS conductor performance specification, Dielectric system design approach, Dewar development for containing phase assemblies, cryo-cooling system design approach, etc. will be described. Finally; overall program schedule, critical milestone events, test plans and progress to date will be reported.

Host factors that promote retrotransposon integration are similar in distantly related eukaryotes

PubMed Central

Rai, Sudhir Kumar; Sangesland, Maya; Lee, Michael; Esnault, Caroline; Cui, Yujin; Chatterjee, Atreyi Ghatak

2017-01-01

Retroviruses and Long Terminal Repeat (LTR)-retrotransposons have distinct patterns of integration sites. The oncogenic potential of retrovirus-based vectors used in gene therapy is dependent on the selection of integration sites associated with promoters. The LTR-retrotransposon Tf1 of Schizosaccharomyces pombe is studied as a model for oncogenic retroviruses because it integrates into the promoters of stress response genes. Although integrases (INs) encoded by retroviruses and LTR-retrotransposons are responsible for catalyzing the insertion of cDNA into the host genome, it is thought that distinct host factors are required for the efficiency and specificity of integration. We tested this hypothesis with a genome-wide screen of host factors that promote Tf1 integration. By combining an assay for transposition with a genetic assay that measures cDNA recombination we could identify factors that contribute differentially to integration. We utilized this assay to test a collection of 3,004 S. pombe strains with single gene deletions. Using these screens and immunoblot measures of Tf1 proteins, we identified a total of 61 genes that promote integration. The candidate integration factors participate in a range of processes including nuclear transport, transcription, mRNA processing, vesicle transport, chromatin structure and DNA repair. Two candidates, Rhp18 and the NineTeen complex were tested in two-hybrid assays and were found to interact with Tf1 IN. Surprisingly, a number of pathways we identified were found previously to promote integration of the LTR-retrotransposons Ty1 and Ty3 in Saccharomyces cerevisiae, indicating the contribution of host factors to integration are common in distantly related organisms. The DNA repair factors are of particular interest because they may identify the pathways that repair the single stranded gaps flanking the sites of strand transfer following integration of LTR retroelements. PMID:29232693

Host factors that promote retrotransposon integration are similar in distantly related eukaryotes.

PubMed

Rai, Sudhir Kumar; Sangesland, Maya; Lee, Michael; Esnault, Caroline; Cui, Yujin; Chatterjee, Atreyi Ghatak; Levin, Henry L

2017-12-01

Retroviruses and Long Terminal Repeat (LTR)-retrotransposons have distinct patterns of integration sites. The oncogenic potential of retrovirus-based vectors used in gene therapy is dependent on the selection of integration sites associated with promoters. The LTR-retrotransposon Tf1 of Schizosaccharomyces pombe is studied as a model for oncogenic retroviruses because it integrates into the promoters of stress response genes. Although integrases (INs) encoded by retroviruses and LTR-retrotransposons are responsible for catalyzing the insertion of cDNA into the host genome, it is thought that distinct host factors are required for the efficiency and specificity of integration. We tested this hypothesis with a genome-wide screen of host factors that promote Tf1 integration. By combining an assay for transposition with a genetic assay that measures cDNA recombination we could identify factors that contribute differentially to integration. We utilized this assay to test a collection of 3,004 S. pombe strains with single gene deletions. Using these screens and immunoblot measures of Tf1 proteins, we identified a total of 61 genes that promote integration. The candidate integration factors participate in a range of processes including nuclear transport, transcription, mRNA processing, vesicle transport, chromatin structure and DNA repair. Two candidates, Rhp18 and the NineTeen complex were tested in two-hybrid assays and were found to interact with Tf1 IN. Surprisingly, a number of pathways we identified were found previously to promote integration of the LTR-retrotransposons Ty1 and Ty3 in Saccharomyces cerevisiae, indicating the contribution of host factors to integration are common in distantly related organisms. The DNA repair factors are of particular interest because they may identify the pathways that repair the single stranded gaps flanking the sites of strand transfer following integration of LTR retroelements.

A New Blondin System for Surveying and Photogrammetry

PubMed Central

Cuesta, Federico; Lopez-Rodriguez, Francisco M.; Esteban, Antonio

2013-01-01

The main objective of the system presented in this paper is to provide surveyors and engineers with a new photogrammetry device that can be easily integrated with surveying total stations and a global navigation satellite system (GNSS) infrastructure at a construction site, taking advantage of their accuracy and overcoming limitations of aerial vehicles with respect to weight, autonomy and skilled operator requirements in aerial photogrammetry. The system moves between two mounting points, in a blondin ropeway configuration, at the construction site, taking pictures and recording the data of the position and the orientation along the cable path. A cascaded extended Kalman filter is used to integrate measurements from the on-board inertial measurement unit (IMU), a GPS and a GNSS. Experimental results taken in a construction site show the system performance, including the validation of the position estimation, with a robotic surveying total station, or the creation of a digital surface model (DSM), using the emergent structure from motion (SfM) techniques and open software. The georeferencing of the DSM is performed based on estimated camera position or using ground control points (GCPs).

Biconic cargo return vehicle with an advanced recovery system. Volume 1: Conceptual design

NASA Technical Reports Server (NTRS)

1990-01-01

The conceptual design of the biconic Cargo Return Vehicle (CRV) is presented. The CRV will be able to meet all of the Space Station Freedom (SSF's) resupply needs. Worth note is the absence of a backup recovery chute in case of Advanced Recovery System (ARS) failure. The high reliability of ram-air parachutes does not warrant the penalty weight that such a system would create on successful missions. The CRV will launch vertically integrated with an Liquid Rocket Booster (LRB) vehicle and meets all NASA restrictions on fuel type for all phases of the mission. Because of the downscaled Orbital Maneuvering Vehicle (OMV) program, the CRV has been designed to be able to transfer cargo by docking directly to the Space Station Freedom as well as with OMV assistance. The CRV will cover enough crossrange to reach its primary landing site, Edwards Airforce Base, and all secondary landing sites with the exception of one orbit. Transportation back to KSC will be via the Boeing Super Guppy. Due to difficulties with man-rating the CRV, it will not be used in a CERV role. A brief summary of the CRV's specifications is given.

Supply Chain Management on IBS Implementation in Klang Valley Construction Industry: Challenges and Issues

NASA Astrophysics Data System (ADS)

Azrizal Fauzi, Mohd; Hasim, Sulaiman; Awang, Anizah; Ridzuan, Ahmad Ruslan Mohd; Nur Yunus, Juzailah

2017-12-01

Industrialized Building System (IBS) is a system where the components of the building are manufactured in a factory and it will be transported to the site to form the structures. The supply chain management (SCM) is a system where the delivery flows of the IBS products from manufacturers to the site. The aim of this research is to identify the major challenges and to analyze the issues on IBS implementation in SCM in Klang Valley from the manufacturers perspective. The methodology used in this paper is based on primary data through questionnaire and interview. Questionnaires were sent to the Manufacturers. It can be concluded that this paper attempts to present more on the challenges and issues that those companies of manufacturers faced during their success journey in finding integration in their supply chain. The main contributions of this paper are integrating all the supply chain integration challenges and issues on IBS. Therefore, these contributions will be helpful for the organization of manufacturers and IBS players that establish the integration in their SCM.

The Emerging Role of the Data Base Manager. Report No. R-1253-PR.

ERIC Educational Resources Information Center

Sawtelle, Thomas K.

The Air Force Logistics Command (AFLC) is revising and enhancing its data-processing capabilities with the development of a large-scale, multi-site, on-line, integrated data base information system known as the Advanced Logistics System (ALS). A data integrity program is to be built around a Data Base Manager (DBM), an individual or a group of…

Perceived Factors Associated with Sustained Improvement Following Participation in a Multicenter Quality Improvement Collaborative.

PubMed

Stone, Sohini; Lee, Henry C; Sharek, Paul J

2016-07-01

The California Perinatal Quality Care Collaborative led the Breastmilk Nutrition Quality Improvement Collaborative from October 2009 to September 2010 to increase the percentage of very low birth weight infants receiving breast milk at discharge in 11 collaborative neonatal ICUs (NICUs). Observed increases in breast milk feeding and decreases in necrotizing enterocolitis persisted for 6 months after the collaborative ended. Eighteen to 24 months after the end of the collaborative, some sites maintained or further increased their gains, while others trended back toward baseline. A study was conducted to assess the qualitative factors that affect sustained improvement following participation. Collaborative leaders at each of the 11 NICUs that participated in the Breastmilk Nutrition Quality Improvement Collaborative were invited to participate in a site-specific one-hour phone interview. Interviews were recorded and transcribed and then analyzed using qualitative research analysis software to identify themes associated with sustained improvement. Eight of 11 invited centers agreed to participate in the interviews. Thematic saturation was achieved by the sixth interview, so further interviews were not pursued. Factors contributing to sustainability included physician involvement within the multidisciplinary teams, continuous education, incorporation of interventions into the daily work flow, and integration of a data-driven feedback system. Early consideration by site leaders of how to integrate best-practice interventions into the daily work flow, and ensuring physician commitment and ongoing education based in continuous data review, should enhance the likelihood of sustaining improvements. To maximize sustained success, future collaborative design should consider proactively identifying and supporting these factors at participating sites.

Examination of Axonal Injury and Regeneration in Microfluidic Neuronal Culture Using Pulsed Laser Microbeam Dissection

PubMed Central

Hellman, Amy N.; Vahidi, Behrad; Kim, Hyung Joon; Mismar, Wael; Steward, Oswald; Jeon, Noo Li; Venugopalan, Vasan

2010-01-01

We describe the integrated use of pulsed laser microbeams and microfluidic cell culture to examine the dynamics of axonal injury and regeneration in vitro. Microfabrication methods are used to place high purity dissociated central nervous system neurons in specific regions that allow the axons to interact with permissive and inhibitory substrates. Acute injury to neuron bundles is produced via the delivery of single 180 ps duration, λ=532 nm laser pulses. Laser pulse energies of 400 nJ and 800 nJ produce partial and complete transection of the axons, respectively, resulting in elliptical lesions 25 μm and 50 μm in size. The dynamics of the resulting degeneration and regrowth of proximal and distal axonal segments are examined for up to 8 h using time-lapse microscopy. We find the proximal and distal dieback distances from the site of laser microbeam irradiation to be roughly equal for both partial and complete transection of the axons. In addition, distinct growth cones emerge from the proximal neurite segments within 1–2 h post-injury, followed by a uniform front of regenerating axons that originate from the proximal segment and traverse the injury site within 8 h. We also examine the use of EGTA to chelate the extracellular calcium and potentially reduce the severity of the axonal degeneration following injury. While we find the addition of EGTA to reduce the severity of the initial dieback, it also hampers neurite repair and interfere with the formation of neuronal growth cones to traverse the injury site. This integrated use of laser microbeam dissection within a microfluidic cell culture system to produce precise zones of neuronal injury shows potential for high-throughput screening of agents to promote neuronal regeneration. PMID:20532390

Conceptual approaches to avian navigation systems.

PubMed

Wallraff, H G

1991-01-01

The general basis of migratory orientation in birds is most probably an endogenous time-and-direction programme. Directions are selected with respect to celestial as well as geomagnetic clues. These clues appear to be integrated within a system that profits from the special advantages of either kind of environmental signal, and thereby can cope with their limitations. Using these clues, and following a genetically determined intended direction (or sequence of directions) over a genetically determined period of time, a bird may reach a larger population-specific area. However, it will hardly be able to find a particular location, such as, for instance, its previous breeding site. Homing to a familiar site over several hundred kilometers of unfamiliar terrain is substantially based on the smelling of atmospheric trace compounds. At shorter distances from home, orientation by means of--presumably visual--familiar landmarks completes the repertoire of mechanisms guiding a bird back home. These mechanisms are considered to be based on different kinds of 'maps' and 'compasses'. Conceptual approaches to the properties of an 'olfactory map' have as yet only reached an early state of speculation.

Integrating family planning and HIV services in western Kenya: the impact on HIV-infected patients' knowledge of family planning and male attitudes toward family planning.

PubMed

Onono, Maricianah; Guzé, Mary A; Grossman, Daniel; Steinfeld, Rachel; Bukusi, Elizabeth A; Shade, Starley; Cohen, Craig R; Newmann, Sara J

2015-01-01

Little information exists on the impact of integrating family planning (FP) services into HIV care and treatment on patients' familiarity with and attitudes toward FP. We conducted a cluster-randomized trial in 18 public HIV clinics with 12 randomized to integrated FP and HIV services and 6 to the standard referral-based system where patients are referred to an FP clinic. Serial cross-sectional surveys were done before (n = 488 women, 486 men) and after (n = 479 women, 481 men) the intervention to compare changes in familiarity with FP methods and attitudes toward FP between integrated and nonintegrated (NI) sites. We created an FP familiarity score based on the number of more effective FP methods patients could identify (score range: 0-6). Generalized estimating equations were used to control for clustering within sites. An increase in mean familiarity score between baseline (mean = 5.16) and post-intervention (mean = 5.46) occurred with an overall mean change of 0.26 (95% confidence intervals [CI] = 0.09, 0.45; p = 0.003) across all sites. At end line, there was no difference in increase of mean FP familiarity scores at intervention versus control sites (mean = 5.41 vs. 5.49, p = 0.94). We observed a relative decrease in the proportion of males agreeing that FP was "women's business" at integrated sites (baseline 42% to end line 30%; reduction of 12%) compared to males at NI sites (baseline 35% to end line 42%; increase of 7%; adjusted odds ration [aOR] = 0.43; 95% CI = 0.22, 0.85). Following FP-HIV integration, familiarity with FP methods increased but did not differ by study arm. Integration was associated with a decrease in negative attitudes toward FP among men.

Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells.

PubMed

Liao, Hsin-Kai; Gu, Ying; Diaz, Arturo; Marlett, John; Takahashi, Yuta; Li, Mo; Suzuki, Keiichiro; Xu, Ruo; Hishida, Tomoaki; Chang, Chan-Jung; Esteban, Concepcion Rodriguez; Young, John; Izpisua Belmonte, Juan Carlos

2015-03-10

To combat hostile viruses, bacteria and archaea have evolved a unique antiviral defense system composed of clustered regularly interspaced short palindromic repeats (CRISPRs), together with CRISPR-associated genes (Cas). The CRISPR/Cas9 system develops an adaptive immune resistance to foreign plasmids and viruses by creating site-specific DNA double-stranded breaks (DSBs). Here we adapt the CRISPR/Cas9 system to human cells for intracellular defense against foreign DNA and viruses. Using HIV-1 infection as a model, our results demonstrate that the CRISPR/Cas9 system disrupts latently integrated viral genome and provides long-term adaptive defense against new viral infection, expression and replication in human cells. We show that engineered human-induced pluripotent stem cells stably expressing HIV-targeted CRISPR/Cas9 can be efficiently differentiated into HIV reservoir cell types and maintain their resistance to HIV-1 challenge. These results unveil the potential of the CRISPR/Cas9 system as a new therapeutic strategy against viral infections.

Infection of human T lymphotropic virus-I-specific immune T cell clones by human T lymphotropic virus-I.

PubMed Central

Mitsuya, H; Jarrett, R F; Cossman, J; Cohen, O J; Kao, C S; Guo, H G; Reitz, M S; Broder, S

1986-01-01

Human T lymphotropic virus-I (HTLV-I)-specific T cell lines were established and cloned. K5, an OKT8+ clone bearing multiple proviral integration sites, retained its HTLV-I-specific cytotoxicity and a normal dependence on interleukin 2 (IL-2), indicating that there is a finite number of transforming integration sites. R2, an OKT4+ HTLV-I-infected clone, initially mounted a proliferative response to HTLV-I; but then its IL-2-independent proliferation increased and the antigen specificity was lost. All HTLV-I-infected clones tested including K7, another OKT8+ transformed cytotoxic clone that had lost its reactivity, expressed comparable levels of T cell receptor beta-chain (TCR-beta) messenger (m)RNA. Although clones K5 and K7 had different functional properties, they had the same rearrangement of the TCR-beta gene, suggesting that they had the same clonal origin. These data indicate that HTLV-I-specific T cells retain their immune reactivity for variable periods of time following infection, but then usually lose it; in some cases, however, no alteration in function can be detected. The data also suggest that different consequences can take place in the same clone depending on the pattern of retroviral infection. Images PMID:2877011

Text Mining Improves Prediction of Protein Functional Sites

PubMed Central

Cohn, Judith D.; Ravikumar, Komandur E.

2012-01-01

We present an approach that integrates protein structure analysis and text mining for protein functional site prediction, called LEAP-FS (Literature Enhanced Automated Prediction of Functional Sites). The structure analysis was carried out using Dynamics Perturbation Analysis (DPA), which predicts functional sites at control points where interactions greatly perturb protein vibrations. The text mining extracts mentions of residues in the literature, and predicts that residues mentioned are functionally important. We assessed the significance of each of these methods by analyzing their performance in finding known functional sites (specifically, small-molecule binding sites and catalytic sites) in about 100,000 publicly available protein structures. The DPA predictions recapitulated many of the functional site annotations and preferentially recovered binding sites annotated as biologically relevant vs. those annotated as potentially spurious. The text-based predictions were also substantially supported by the functional site annotations: compared to other residues, residues mentioned in text were roughly six times more likely to be found in a functional site. The overlap of predictions with annotations improved when the text-based and structure-based methods agreed. Our analysis also yielded new high-quality predictions of many functional site residues that were not catalogued in the curated data sources we inspected. We conclude that both DPA and text mining independently provide valuable high-throughput protein functional site predictions, and that integrating the two methods using LEAP-FS further improves the quality of these predictions. PMID:22393388