New directions and challenges for the O ampersand M committee

International Nuclear Information System (INIS)

Pelletier, J.P.

1996-01-01

In the second half of the decade of the nineties, the imperative of finding new ways to improve efficiency while maintaining safety is taking on a new urgency. The looming deregulation of the electric industry and the expected competition in the power production business fuels this urgency. The recently completed ASME research in the area of Risk-Based Inservice Testing offers the Operations and Maintenance (O ampersand M) Committee an opportunity to meet this challenge. This opportunity, however, offers its own challenges. New ways of thinking about inservice testing and new technical skills will be needed to successfully incorporate this technology into Code documents. It will not be adequate to rely solely on incorporation of risk-based methods into the Code to meet this challenge. A thorough self assessment of what O ampersand M does, how it is done, and the value added will help assure that the directions which are pursued do, indeed, meet the challenge ahead

Tailor-Made Protein Tyrosine Phosphatases: In Vitro Site-Directed Mutagenesis of PTEN and PTPRZ-B

NARCIS (Netherlands)

Luna, S.; Mingo, J.; Aurtenetxe, O.; Blanco, L.; Amo, L.; Schepens, J.; Hendriks, W.J.A.J.; Pulido, R.

2016-01-01

In vitro site-directed mutagenesis (SDM) of protein tyrosine phosphatases (PTPs) is a commonly used approach to experimentally analyze PTP functions at the molecular and cellular level and to establish functional correlations with PTP alterations found in human disease. Here, using the

HIV vaccines: current challenges and future directions.

Science.gov (United States)

Avrett, Sam; Collins, Chris

2002-07-01

Volume seven of the Review will mark the tenth anniversary of the Canadian HIV/AIDS Legal Network with a series of articles that describe past developments and future directions in several areas of policy and law related to HIV/AIDS. The following article is the first of these, discussing current challenges and future directions in the development of and access to HIV vaccines. It argues that governments are under public health, ethical, and legal obligations to develop and provide access to HIV vaccines. It further explains what is required for governments to fulfill their obligations: additional commitment and resources for HIV vaccine development in the context of increased global research and development regarding diseases of the poor; increased support and advocacy for partnerships to develop HIV vaccines; enhanced regulatory capacity in every country to review, approve, and monitor HIV vaccines; and assurance of global supply of, procurement of, delivery of, and access to vaccines in the context of efforts to increase global access to public health measures and technologies.

Directing Matter and Energy: Five Challenges for Science and the Imagination

Energy Technology Data Exchange (ETDEWEB)

Hemminger, J.; Fleming, G.; Ratner, M.

2007-12-20

The twin aspects of energy and control (or direction) are the underlying concepts. Matter and energy are closely linked, and their understanding and control will have overwhelming importance for our civilization, our planet, our science, and our technology. This importance ranges even beyond the large portfolio of BES, both because these truly significant Grand Challenges confront many other realms of science and because even partial solutions to these challenges will enrich scientists’ collective imagination and ability to solve problems with new ideas and new methods.

In vitro propagation of Cymbidium goeringii Reichenbach fil. through direct adventitious shoot regeneration.

Science.gov (United States)

Park, Han Yong; Kang, Kyung Won; Kim, Doo Hwan; Sivanesan, Iyyakkannu

2018-03-01

The influence of 2,4-dichlorophenoxyacetic acid (2,4-D), benzyladenine (BA), and thidiazuron (TDZ) on direct rhizome induction and shoot formation from rhizome explants of Cymbidium goeringii was explored. Rhizome segments obtained from in vitro seed cultures of C. goeringii were placed on Murashige and Skoog (MS) medium incorporated with 5, 10, 20, or 40 µM 2,4-D and 1, 2, 4, or 8 µM BA or TDZ alone or in combination with 20 µM 2,4-D. The explants developed only rhizomes on MS medium with or without 2,4-D. The highest percent of rhizome formation (100%) was obtained on MS medium incorporated with 20 μM of 2,4-D. The morphology and number of rhizomes varied with the level of 2,4-D in the medium. Direct adventitious shoot formation was achieved on medium incorporated with BA or TDZ. The adventitious shoots produced per explant significantly increased with the supplementation of 2,4-D to cytokinin-containing medium. The highest mean of 21.8 ± 1.8 shoot buds per rhizome segment was obtained in medium fortified with 20 μM 2,4-D and 2 μM TDZ. The greatest percent of root induction (100%) and the mean of 5.3 ± 1.1 roots per shoot were achieved on ½ MS medium incorporated with 2 μM of α-naphthaleneacetic acid. About 97% of the in vitro-produced plantlets acclimatized in the greenhouse. An efficient in vitro propagation protocol was thus developed for C. goeringii using rhizome explants.

In Vitro Study of Directly Bioprinted Perfusable Vasculature Conduits.

Science.gov (United States)

Zhang, Yahui; Yu, Yin; Akkouch, Adil; Dababneh, Amer; Dolati, Farzaneh; Ozbolat, Ibrahim T

2015-01-01

The ability to create three dimensional (3D) thick tissues is still a major tissue engineering challenge. It requires the development of a suitable vascular supply for an efficient media exchange. An integrated vasculature network is particularly needed when building thick functional tissues and/or organs with high metabolic activities, such as the heart, liver and pancreas. In this work, human umbilical vein smooth muscle cells (HUVSMCs) were encapsulated in sodium alginate and printed in the form of vasculature conduits using a coaxial deposition system. Detailed investigations were performed to understand the dehydration, swelling and degradation characteristics of printed conduits. In addition, because perfusional, permeable and mechanical properties are unique characteristics of natural blood vessels, for printed conduits these properties were also explored in this work. The results show that cells encapsulated in conduits had good proliferation activities and that their viability increased during prolonged in vitro culture. Deposition of smooth muscle matrix and collagen was observed around the peripheral and luminal surface in long-term cultured cellular vascular conduit through histology studies.

Paired assessment of volatile anesthetic concentrations with synaptic actions recorded in vitro.

Directory of Open Access Journals (Sweden)

Stuart J McDougall

Full Text Available The volatile anesthetic isoflurane poses a number of experimental challenges in the laboratory. Due to its rapid evaporation, the open conditions of most in vitro electrophysiological recording systems make the determination of actual isoflurane concentrations a challenge. Since the absolute anesthetic concentration in solution is directly related to efficacy, concentration measurements are important to allow comparisons between laboratory and clinical studies. In this study we quantify the sources of isoflurane loss during experimentation and describe a method for the measurement of isoflurane concentrations using gas chromatography and mass spectrometry simultaneous to in vitro electrophysiological measurements. Serial samples of perfused bath solution allowed correlation of isoflurane concentrations with ongoing biological effects. Saturated physiological solutions contained 13.4 +/- 0.2 mM isoflurane and were diluted to desired "nominal" concentrations for experiments. The perfusion system established stable isoflurane concentrations within the bath by 2 minutes. However, bath isoflurane concentrations varied substantially and unpredictably between experiments. The magnitudes of such discrepancies in isoflurane concentrations spanned clinically important levels. Our studies suggest that, despite countermeasures, solution handling significantly impacted the isoflurane content in the tissue bath. The magnitude of these discrepancies appears to necessitate systematic direct measurement of bath isoflurane concentrations during most in vitro conditions.

Key Challenges and Future Directions for Educational Research on Scientific Argumentation

Science.gov (United States)

Henderson, J. Bryan; McNeill, Katherine L.; González-Howard, María; Close, Kevin; Evans, Mat

2018-01-01

At the 2015 "NARST: A Worldwide Organization for Improving Science Teaching and Learning Through Research" Annual International Conference, a group of scholars held an extended pre-conference workshop to discuss key challenges and future directions faced by argumentation researchers around the world. This wide-ranging group of…

Geospatial Brokering - Challenges and Future Directions

Science.gov (United States)

White, C. E.

2012-12-01

An important feature of many brokers is to facilitate straightforward human access to scientific data while maintaining programmatic access to it for system solutions. Standards-based protocols are critical for this, and there are a number of protocols to choose from. In this discussion, we will present a web application solution that leverages certain protocols - e.g., OGC CSW, REST, and OpenSearch - to provide programmatic as well as human access to geospatial resources. We will also discuss managing resources to reduce duplication yet increase discoverability, federated search solutions, and architectures that combine human-friendly interfaces with powerful underlying data management. The changing requirements witnessed in brokering solutions over time, our recent experience participating in the EarthCube brokering hack-a-thon, and evolving interoperability standards provide insight to future technological and philosophical directions planned for geospatial broker solutions. There has been much change over the past decade, but with the unprecedented data collaboration of recent years, in many ways the challenges and opportunities are just beginning.

Validation of Alternative In Vitro Methods to Animal Testing: Concepts, Challenges, Processes and Tools.

Science.gov (United States)

Griesinger, Claudius; Desprez, Bertrand; Coecke, Sandra; Casey, Warren; Zuang, Valérie

This chapter explores the concepts, processes, tools and challenges relating to the validation of alternative methods for toxicity and safety testing. In general terms, validation is the process of assessing the appropriateness and usefulness of a tool for its intended purpose. Validation is routinely used in various contexts in science, technology, the manufacturing and services sectors. It serves to assess the fitness-for-purpose of devices, systems, software up to entire methodologies. In the area of toxicity testing, validation plays an indispensable role: "alternative approaches" are increasingly replacing animal models as predictive tools and it needs to be demonstrated that these novel methods are fit for purpose. Alternative approaches include in vitro test methods, non-testing approaches such as predictive computer models up to entire testing and assessment strategies composed of method suites, data sources and decision-aiding tools. Data generated with alternative approaches are ultimately used for decision-making on public health and the protection of the environment. It is therefore essential that the underlying methods and methodologies are thoroughly characterised, assessed and transparently documented through validation studies involving impartial actors. Importantly, validation serves as a filter to ensure that only test methods able to produce data that help to address legislative requirements (e.g. EU's REACH legislation) are accepted as official testing tools and, owing to the globalisation of markets, recognised on international level (e.g. through inclusion in OECD test guidelines). Since validation creates a credible and transparent evidence base on test methods, it provides a quality stamp, supporting companies developing and marketing alternative methods and creating considerable business opportunities. Validation of alternative methods is conducted through scientific studies assessing two key hypotheses, reliability and relevance of the

Effect-directed analysis: Current status and future challenges

Science.gov (United States)

Hong, Seongjin; Giesy, John P.; Lee, Jung-Suk; Lee, Jong-Hyeon; Khim, Jong Seong

2016-09-01

Effect-directed analysis (EDA) has become useful for identification of toxicant(s) that occur in mixtures in the environment, especially those that are causative agents of specific adverse effects. Here, we summarize and review EDA methodology including preparation of samples, biological analyses, fractionations, and instrumental analyses, highlighting key scientific advancements. A total of 63 documents since 1999 (Scopus search) including 46 research articles, 13 review papers, and 4 project descriptions, have been collected and reviewed in this study. At the early stage (1999-2010), most studies that applied EDA focused on organic extracts of freshwater and coastal contaminated sediments and wastewater. Toxic effects were often measured using cell-based bioassays ( in vitro) and the causative chemicals were identified by use of low resolution gas chromatography with mass selective detector (GCMSD). More recently (2010-present), EDA has been extended to various matrices such as biota, soil, crude oil, and suspended solids and techniques have been improved to include determination of bioavailability in vivo. In particular, methods for non-target screenings of organic chemicals in environmental samples using cutting-edge instrumentation such as time of flight-mass spectrometry (ToF-MS), Fourier transform-ion cyclotron resonance (FT-ICR), and Orbitrap mass spectrometer have been developed. This overview provides descriptions of recent improvements of EDA and suggests future research directions based on current understandings and limitations.

Direct bacterial killing in vitro by recombinant Nod2 is compromised by Crohn's disease-associated mutations.

Directory of Open Access Journals (Sweden)

Laurent-Herve Perez

2010-06-01

Full Text Available A homeostatic relationship with the intestinal microflora is increasingly appreciated as essential for human health and wellbeing. Mutations in the leucine-rich repeat (LRR domain of Nod2, a bacterial recognition protein, are associated with development of the inflammatory bowel disorder, Crohn's disease. We investigated the molecular mechanisms underlying disruption of intestinal symbiosis in patients carrying Nod2 mutations.In this study, using purified recombinant LRR domains, we demonstrate that Nod2 is a direct antimicrobial agent and this activity is generally deficient in proteins carrying Crohn's-associated mutations. Wild-type, but not Crohn's-associated, Nod2 LRR domains directly interacted with bacteria in vitro, altered their metabolism and disrupted the integrity of the plasma membrane. Antibiotic activity was also expressed by the LRR domains of Nod1 and other pattern recognition receptors suggesting that the LRR domain is a conserved anti-microbial motif supporting innate cellular immunity.The lack of anti-bacterial activity demonstrated with Crohn's-associated Nod2 mutations in vitro, supports the hypothesis that a deficiency in direct bacterial killing contributes to the association of Nod2 polymorphisms with the disease.

Challenges in Design of an Orientation free Micro Direct Methanol Fuel Cell (µDMFC)

DEFF Research Database (Denmark)

Omidvarnia, Farzaneh; Hansen, Hans Nørgaard; Hales, Jan Harry

2014-01-01

the challenges in design and manufacturing of a micro direct methanol fuel cell (μDMFC) as the power generator in hearing aid devices is investigated. Among the different challenges in design for μDMFC, the CO2 bubble management and orientation independency of the cell are addressed by proposing a spring loaded...

Cascading Events, Technology and the Floods Directive: future challenges

Directory of Open Access Journals (Sweden)

Pescaroli Gianluca

2016-01-01

Full Text Available Cascading events can be referred to multidimensional disasters, where a primary trigger generates a nonlinear series of secondary emergencies that escalate in time, becoming eventually the priority to tackle. In this process, critical infrastructure can be handled as roots of vulnerabilities, because they accumulate both physical attributes and functional nodes. When compromised, they produce widespread breakdowns of society, but also orient emergency responses and long-term recovery. Although floods have been widely associated to the failure of vulnerable equipments or to the disruption of strategic sectors such as energy, communication and transportation, their integration with the emerging concept of cascading has been limited. This open topic presents many challenges for scholars, researchers and practitioners, in particular when the implementation of the EU Floods Directive is considered. The paper presents an overview of the Floods Directive and its relation with the cascading events, using case studies and examples from the existing literature to point out missing links and gaps in the legislation. Conclusions argue that the Directive considers only local geographical scales and limited temporal horizons, which can be result inadequate to limit the escalation of events.

Immunoglobulin with High-Titer In Vitro Cross-Neutralizing Hepatitis C Virus Antibodies Passively Protects Chimpanzees from Homologous, but Not Heterologous, Challenge

DEFF Research Database (Denmark)

Bukh, Jens; Engle, Ronald E.; Faulk, Kristina

2015-01-01

The importance of neutralizing antibodies (NAbs) in protection against hepatitis C virus (HCV) remains controversial. We infused a chimpanzee with H06 immunoglobulin from a genotype 1a HCV-infected patient and challenged with genotype strains efficiently neutralized by H06 in vitro. Genotype 1a...

Challenges of a pharmacist-directed peer support program among adolescents with diabetes.

Science.gov (United States)

Sims, Lisa M; Haines, Seena L

2011-01-01

To describe the development and challenges of a pharmacist-directed peer support program among adolescents with diabetes. The program was designed as adjunctive therapy for adolescents receiving care at the Diabetes Education and Research Center (DERC). DERC is an interdisciplinary facility at which the clinical pharmacist provides direct, diabetes-related patient care services. Through collaboration with DERC, pharmacists developed and facilitated each component of the program. The U. S. Diabetes Conversation Map program was used for the educational component of the program. This is an innovative, American Diabetes Association-approved tool for providing group education on diabetes self-management. As trained facilitators of the conversation maps, the investigators educated group leaders on how to be facilitators for their peers. Glycosylated hemoglobin (A1C), diabetes self-management skills, and diabetes-related quality of life were measured at baseline and following program completion. Qualitative outcomes were measured via validated questionnaires in adolescents with type 1 diabetes. The investigators facilitated the gathering of all quantitative and qualitative data. Several participants did not meet all inclusion criteria; therefore, data from only six participants were assessed. A1C increased among participants, affirming the challenge of metabolic control during adolescence. Despite this, qualitative analysis of questionnaires revealed improvements in adherence to lifestyle modifications and health perception after program completion. Evidence illustrates beneficial effects of peer-facilitated support on the physical and psychological challenges of diabetes self-management; however, challenges existed when implementing this program among adolescents. Suggestions to overcome these challenges include same-sex support groups, use of an appealing setting for participants, incorporation into camps or after-school programs, and extended program length

Production of biofuels and biochemicals by in vitro synthetic biosystems: Opportunities and challenges.

Science.gov (United States)

Zhang, Yi-Heng Percival

2015-11-15

The largest obstacle to the cost-competitive production of low-value and high-impact biofuels and biochemicals (called biocommodities) is high production costs catalyzed by microbes due to their inherent weaknesses, such as low product yield, slow reaction rate, high separation cost, intolerance to toxic products, and so on. This predominant whole-cell platform suffers from a mismatch between the primary goal of living microbes - cell proliferation and the desired biomanufacturing goal - desired products (not cell mass most times). In vitro synthetic biosystems consist of numerous enzymes as building bricks, enzyme complexes as building modules, and/or (biomimetic) coenzymes, which are assembled into synthetic enzymatic pathways for implementing complicated bioreactions. They emerge as an alternative solution for accomplishing a desired biotransformation without concerns of cell proliferation, complicated cellular regulation, and side-product formation. In addition to the most important advantage - high product yield, in vitro synthetic biosystems feature several other biomanufacturing advantages, such as fast reaction rate, easy product separation, open process control, broad reaction condition, tolerance to toxic substrates or products, and so on. In this perspective review, the general design rules of in vitro synthetic pathways are presented with eight supporting examples: hydrogen, n-butanol, isobutanol, electricity, starch, lactate,1,3-propanediol, and poly-3-hydroxylbutyrate. Also, a detailed economic analysis for enzymatic hydrogen production from carbohydrates is presented to illustrate some advantages of this system and the remaining challenges. Great market potentials will motivate worldwide efforts from multiple disciplines (i.e., chemistry, biology and engineering) to address the remaining obstacles pertaining to cost and stability of enzymes and coenzymes, standardized building parts and modules, biomimetic coenzymes, biosystem optimization, and scale

Connectivity inference from neural recording data: Challenges, mathematical bases and research directions.

Science.gov (United States)

Magrans de Abril, Ildefons; Yoshimoto, Junichiro; Doya, Kenji

2018-06-01

This article presents a review of computational methods for connectivity inference from neural activity data derived from multi-electrode recordings or fluorescence imaging. We first identify biophysical and technical challenges in connectivity inference along the data processing pipeline. We then review connectivity inference methods based on two major mathematical foundations, namely, descriptive model-free approaches and generative model-based approaches. We investigate representative studies in both categories and clarify which challenges have been addressed by which method. We further identify critical open issues and possible research directions. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Cell sources for in vitro human liver cell culture models

Science.gov (United States)

Freyer, Nora; Damm, Georg; Seehofer, Daniel; Knöspel, Fanny

2016-01-01

In vitro liver cell culture models are gaining increasing importance in pharmacological and toxicological research. The source of cells used is critical for the relevance and the predictive value of such models. Primary human hepatocytes (PHH) are currently considered to be the gold standard for hepatic in vitro culture models, since they directly reflect the specific metabolism and functionality of the human liver; however, the scarcity and difficult logistics of PHH have driven researchers to explore alternative cell sources, including liver cell lines and pluripotent stem cells. Liver cell lines generated from hepatomas or by genetic manipulation are widely used due to their good availability, but they are generally altered in certain metabolic functions. For the past few years, adult and pluripotent stem cells have been attracting increasing attention, due their ability to proliferate and to differentiate into hepatocyte-like cells in vitro. However, controlling the differentiation of these cells is still a challenge. This review gives an overview of the major human cell sources under investigation for in vitro liver cell culture models, including primary human liver cells, liver cell lines, and stem cells. The promises and challenges of different cell types are discussed with a focus on the complex 2D and 3D culture approaches under investigation for improving liver cell functionality in vitro. Finally, the specific application options of individual cell sources in pharmacological research or disease modeling are described. PMID:27385595

Beneficial effect of directional freezing on in vitro viability of cryopreserved sheep whole ovaries and ovarian cortical slices.

Science.gov (United States)

Maffei, S; Pennarossa, G; Brevini, T A L; Arav, A; Gandolfi, F

2014-01-01

Does directional freezing improve the structural and functional integrity of ovarian fragments compared with conventional slow freezing and to whole ovary cryopreservation? Compared with slow freezing, the use of directional freezing significantly improves all structural and functional parameters of ovarian fragments assessed in vitro and, overall, whole ovaries were better preserved than ovarian fragments. Directional freezing has been developed to provide an alternative way to cryopreserve large biological samples and it is known to improve the structural and functional integrity of whole ovaries. Conventional slow freezing of ovarian fragments is the procedure more widely used in clinical settings but it causes substantial structural damage that limits the functional period after transfer back into the patient. We performed a 2 × 2 factorial design experiment on a total of 40 sheep ovaries, divided into four groups (n = 10 ovaries per group): (i) directional freezing of whole ovary (DFwo); (ii) directional freezing of ovarian fragments (DFof); (iii) conventional freezing of whole ovary (CFwo); (iv) conventional freezing of ovarian fragments (CFof). An additional eight ovaries were used as fresh controls. Ewe ovaries were randomly assigned to one of the experimental groups and frozen accordingly. Upon thawing, ovarian tissue was examined morphologically and cultured in vitro for 7 days. Samples were analyzed for cell proliferation and apoptosis, for DNA damage and repair activity, and for the presence of a panel of heat shock proteins (HSPs) by immunohistochemistry. Most studied parameters were significantly improved (P sheep ovaries, which are smaller than human ovaries and therefore may withstand the procedures better. Improved integrity of ovarian morphology may translate to improved outcomes after transplantation. Alternatively, the particularly good preservation of whole ovaries suggests they could provide a source of ovarian follicles for in vitro culture

Bacterial challenge of NISSHO ultrafilter ETF 609: results of in vitro testing.

Science.gov (United States)

Krautzig, S; Lonnemann, G; Shaldon, S; Koch, K M

1996-07-01

In hemodialysis, a certain degree of bacterial contamination on the dialysate side is a regular finding. Concern has been growing that this contamination may lead to a chronic inflammatory response in the patient. Ultrafiltration of dialysate can be used to reduce bacterial content and levels of cytokine-inducing substances upstream of the patient's dialyzer. The aim of this study was to test in vitro the rejection capacity of a polysulfone hollow-fiber ultrafilter (ETF 609, NISSHO Co., Osaka, Japan) challenged with bacterial filtrates derived from Pseudomonas aeruginosa PA103. Results showed a reduction of interleukin-1 beta-inducing activity (measured on peripheral blood mononuclear cells) from 5,035 +/- 394 pg/ml prefilter to nondetectable levels postfilter and endotoxin levels (limulus amebocyte lysate assay) of 4,167 +/- 1,079 versus 12 +/- 2 pg/ml, respectively. In conclusion, ultrafiltration of dialysate with the polysulfone ultrafilter ETF 609 leads to a potent reduction of cytokine-inducing activity.

Human epithelial cells increase their rigidity with ageing in vitro: direct measurements

International Nuclear Information System (INIS)

Berdyyeva, Tamara K; Woodworth, Craig D; Sokolov, Igor

2005-01-01

The decrease in elasticity of epithelial tissues with ageing contributes to many human diseases. This change was previously attributed to increased crosslinking of extracellular matrix proteins. Here we show that individual human epithelial cells also become significantly more rigid during ageing in vitro. Using atomic force microscopy (AFM), we found that the Young's modulus of viable cells was consistently increased two- to four-fold in older versus younger cells. Direct visualization of the cytoskeleton using a novel method involving the AFM suggested that increased rigidity of ageing cells was due to a higher density of cytoskeletal fibres. Our results identify a unique mechanism that might contribute to the age-related loss of elasticity in epithelial tissues

e-Business challenges and directions: important themes from the first ICE-B workshop

NARCIS (Netherlands)

Marca, David; Bulander, Rebecca; Kruslin, Cornelia; Shishkov, Boris; van Sinderen, Marten J.; Obaidat, Mohammad S.; Tsihrintzis, George A.; Filipe, Joaquim

2012-01-01

A three-day asynchronous, interactive workshop was held at ICE-B’10 in Piraeus, Greece in July of 2010. This event captured conference themes for e-Business challenges and directions across four subject areas: a) e-Business applications and models, b) enterprise engineering, c) mobility, d) business

Implementing the European Marine Strategy Framework Directive: Scientific challenges and opportunities

Science.gov (United States)

Newton, Alice; Borja, Angel; Solidoro, Cosimo; Grégoire, Marilaure

2015-10-01

The Marine Strategy Framework Directive (MSFD; EC, 2008) is an ambitious European policy instrument that aims to achieve Good Environmental Status (GES) in the 5,720,000 km2 of European seas by 2020, using an Ecosystem Approach. GES is to be assessed using 11 descriptors and up to 56 indicators (European Commission, 2010), and the goal is for clean, healthy and productive seas that are the basis for marine-based development, known as Blue-Growth. The MSFD is one of many policy instruments, such as the Water Framework Directive, the Common Fisheries Policy and the Habitats Directive that, together, should result in "Healthy Oceans and Productive Ecosystems - HOPE". Researchers working together with stakeholders such as the Member States environmental agencies, the European Environmental Agency, and the Regional Sea Conventions, are to provide the scientific knowledge basis for the implementation of the MSFD. This represents both a fascinating challenge and a stimulating opportunity.

Chinese foreign direct investment in Brazil: motivations, challenges and prospects

Directory of Open Access Journals (Sweden)

Yanyi Wang

2013-08-01

Full Text Available The objective of this research was to identify motivations presented as established by Chinese companies that perform foreign direct investment in Brazil, as well as the challenges encountered in this process and their future prospects. After a structured bibliography review of topics from foreign direct investment and Chinese investment in Latin America and Brazil, we conducted an empirical study on qualitative basis from the consultation to key informants, the methodological approach that has been used in international business studies that focus on containing China or with managers and / or Chinese managers as respondents. The procedures of data collection involved semi-structured interviews conducted, in person and by phone, in addition to questionnaires containing open questions, while processing the data followed the protocol of classical content analysis. The results of the empirical stage of the research highlight cultural and political issues that underlie strategic business decisions.

Social determinants of health in selected slum areas in Jordan: challenges and policy directions.

Science.gov (United States)

Ajlouni, Musa T

2016-01-01

The unplanned urbanization in Jordan has over time created many informal settlements "slums" around big cities as Amman, Zerka and Aqaba. The purpose of this study was to highlight the most common challenges related to social determinants of health in two selected slum areas in Amman and Aqaba and suggest policy directions and interventions to meet these challenges. In addition to a prestructured interview with all household heads living in the two slum sites, focus group meetings with a purposefully selected sample of 12 slum dwellers in each site were used to assess the structural and intermediary determinants of health as perceived by slum residents in the two study locations. The study found that slum residents in the two locations suffer from many challenges as severe poverty; unemployment; illiteracy and low education attainments; gender discrimination; insufficient and poor diet; social and official exclusion; unhealthy environment; lack of water supply, electricity and basic sanitation facilities; high prevalence of diseases; and insufficient and inappropriate health services. Specific policy directions to meet these challenges were recommended and grouped into three main clusters: social protection, social inclusion and empowerment. New plans and tools should be developed by local authorities in Jordan to understand, protect, include and empower those vulnerable people who are forced to live in these unhealthy and inhuman environments. Copyright © 2014 John Wiley & Sons, Ltd.

In vitro flow cytometry-based screening platform for cellulase engineering

Science.gov (United States)

Körfer, Georgette; Pitzler, Christian; Vojcic, Ljubica; Martinez, Ronny; Schwaneberg, Ulrich

2016-01-01

Ultrahigh throughput screening (uHTS) plays an essential role in directed evolution for tailoring biocatalysts for industrial applications. Flow cytometry-based uHTS provides an efficient coverage of the generated protein sequence space by analysis of up to 107 events per hour. Cell-free enzyme production overcomes the challenge of diversity loss during the transformation of mutant libraries into expression hosts, enables directed evolution of toxic enzymes, and holds the promise to efficiently design enzymes of human or animal origin. The developed uHTS cell-free compartmentalization platform (InVitroFlow) is the first report in which a flow cytometry-based screened system has been combined with compartmentalized cell-free expression for directed cellulase enzyme evolution. InVitroFlow was validated by screening of a random cellulase mutant library employing a novel screening system (based on the substrate fluorescein-di-β-D-cellobioside), and yielded significantly improved cellulase variants (e.g. CelA2-H288F-M1 (N273D/H288F/N468S) with 13.3-fold increased specific activity (220.60 U/mg) compared to CelA2 wildtype: 16.57 U/mg). PMID:27184298

Issues in vaccinology: Present challenges and future directions.

Science.gov (United States)

Sheerin, Dylan; Openshaw, Peter Jm; Pollard, Andrew J

2017-12-01

Vaccination is a principal and highly cost-effective means of controlling infectious diseases, providing direct protection against pathogens by conferring long-lasting immunological memory and inducing population-level herd immunity. Despite rapid ongoing progress in vaccinology, there remain many obstacles to the development and deployment of novel or improved vaccines; these include the underlying science of how to induce and sustain appropriate protective immune responses as well as bureaucratic, logistic and socio-political hurdles. The failure to distribute and administer existing vaccines to at-risk communities continues to account for a large proportion of infant mortality worldwide: almost 20 million children do not have access to basic vaccines and several million still die each year as a result. While emerging epidemic or pandemic diseases pose a significant threat to global health and prosperity, there are many infectious diseases which provide a continuous or cyclical burden on healthcare systems which also need to be addressed. Gaps in knowledge of the human immune system stand in the way of developing technologies to overcome individual and pathogenic variation. The challenges in tackling infectious disease and directions that the field of preventive medicine may take to improve the current picture of global health are the focus of this review. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cholinergic neuromodulation controls directed temporal communication in neocortex in vitro

Directory of Open Access Journals (Sweden)

Anita K Roopun

2010-03-01

Full Text Available Acetylcholine is the primary neuromodulator involved in cortical arousal in mammals. Cholinergic modulation is involved in conscious awareness, memory formation and attention – processes that involve intercommunication between different cortical regions. Such communication is achieved in part through temporal structuring of neuronal activity by population rhythms, particularly in the beta and gamma frequency ranges (12 – 80 Hz. Here we demonstrate, using in vitro and in silico models, that spectrally identical patterns of beta2 and gamma rhythms are generated in primary sensory areas and polymodal association areas by fundamentally different local circuit mechanisms: Glutamatergic excitation induced beta2 frequency population rhythms only in layer 5 association cortex whereas cholinergic neuromodulation induced this rhythm only in layer 5 primary sensory cortex. This region-specific sensitivity of local circuits to cholinergic modulation allowed for control of the extent of cortical temporal interactions. Furthermore, the contrasting mechanisms underlying these beta2 rhythms produced a high degree of directionality, favouring an influence of association cortex over primary auditory cortex.

Cholinergic Neuromodulation Controls Directed Temporal Communication in Neocortex in Vitro

Science.gov (United States)

Roopun, Anita K.; LeBeau, Fiona E.N.; Rammell, James; Cunningham, Mark O.; Traub, Roger D.; Whittington, Miles A.

2010-01-01

Acetylcholine is the primary neuromodulator involved in cortical arousal in mammals. Cholinergic modulation is involved in conscious awareness, memory formation and attention – processes that involve intercommunication between different cortical regions. Such communication is achieved in part through temporal structuring of neuronal activity by population rhythms, particularly in the beta and gamma frequency ranges (12–80 Hz). Here we demonstrate, using in vitro and in silico models, that spectrally identical patterns of beta2 and gamma rhythms are generated in primary sensory areas and polymodal association areas by fundamentally different local circuit mechanisms: Glutamatergic excitation induced beta2 frequency population rhythms only in layer 5 association cortex whereas cholinergic neuromodulation induced this rhythm only in layer 5 primary sensory cortex. This region-specific sensitivity of local circuits to cholinergic modulation allowed for control of the extent of cortical temporal interactions. Furthermore, the contrasting mechanisms underlying these beta2 rhythms produced a high degree of directionality, favouring an influence of association cortex over primary auditory cortex. PMID:20407636

Membrane-based processes for wastewater nutrient recovery: Technology, challenges, and future direction.

Science.gov (United States)

Xie, Ming; Shon, Ho Kyong; Gray, Stephen R; Elimelech, Menachem

2016-02-01

Wastewater nutrient recovery holds promise for more sustainable water and agricultural industries. We critically review three emerging membrane processes - forward osmosis (FO), membrane distillation (MD) and electrodialysis (ED) - that can advance wastewater nutrient recovery. Challenges associated with wastewater nutrient recovery were identified. The advantages and challenges of applying FO, MD, and ED technologies to wastewater nutrient recovery are discussed, and directions for future research and development are identified. Emphasis is given to exploration of the unique mass transfer properties of these membrane processes in the context of wastewater nutrient recovery. We highlight that hybridising these membrane processes with existing nutrient precipitation process will lead to better management of and more diverse pathways for near complete nutrient recovery in wastewater treatment facilities. Copyright © 2015 Elsevier Ltd. All rights reserved.

IPTV: Challenges and Future Directions

NARCIS (Netherlands)

O.M. Bonastre; M.J. Montpetit; P.S. Cesar Garcia (Pablo Santiago)

2011-01-01

htmlabstractThe move to IP Protocol Television (IPTV) has challenged the traditional television industry by opening the Internet to high quality real time television content delivery. Thus it has provided an enabling set of key technologies to understand and foster further innovations in the

In vivo gluten challenge in coeliac disease

Directory of Open Access Journals (Sweden)

HJ Ellis

2001-01-01

Full Text Available In vivo gluten challenge has been used since the early 1950s to study the role of cereal fractions in celiac disease. While early studies relied on crude indicators of celiac toxicity, the advent of jejunal biopsy and sophisticated immunohistochemical techniques has allowed accurate studies to be performed. Studies to determine the nature of the cereal component that is toxic to patients with celiac disease have concentrated on wheat because of its nutritional importance. A number of in vitro studies indicated the presence of one or more celiac-activating epitopes with the N-terminus of the A-gliadin molecule. In vivo challenge with three synthetic peptides subsequently indicated the toxicity of a peptide corresponding to amino acids 31 to 49 of A-gliadin. In vivo gluten challenge is the gold standard for the assessment of celiac toxicity; however, jejunal biopsy is a relatively invasive procedure, thus, other methods have been investigated. Direct infusion of the rectum with gluten has been shown to result in an increase in mucosal intraepithelial lymphocytes, occurring only in celiac patients. This method has been used to study the celiac toxicity of gliadin subfractions. The in vitro technique of small intestinal biopsy organ culture is also a useful tool and appears to give the same results as in vivo challenge. The importance of tiny amounts of gliadin in the diet, such as that which occurs in wheat starch, has been studied by in vivo challenge; this technique has clarified the position of oats in the gluten-free diet. Several studies suggest that this cereal may be included in the diet of most adult celiac patients. Studies of the transport of gliadin across the enterocyte following ingestion or challenge suggest that gliadin may be metabolized by a different pathway in celiac disease. This could result in an abnormal presentation to the immune system, triggering a pathogenic rather than a tolerogenic response.

Direct 99mTc labeling of monoclonal antibodies: radiolabeling and in vitro stability

International Nuclear Information System (INIS)

Garron, J.Y.; Moinereau, M.; Pasqualini, R.; Saccavini, J.C.

1991-01-01

Direct labeling involves 99m Tc binding to different donor groups on the protein, giving multiple binding sites of various affinities resulting in an in vivo instability. The stability has been considerably improved by activating the antibody using a controlled reduction reaction (using 2-aminoethanethiol). This reaction generates sulfhydryl groups, which are known to strongly bind 99m Tc. The direct 99m Tc antibody labeling method was explored using whole antibodies and fragments. Analytical methods were developed for routine evaluation of radiolabeling yield and in vitro stability. Stable direct antibody labeling with 99m Tc requires the generation of sulfhydryl groups, which show high affinity binding sites for 99m Tc. Such groups are obtained with 2-aminoethanethiol (AET), which induces the reduction of the intrachain or interchain disulfide bond, with no structural deterioration or any loss of immunobiological activity of the antibody. The development of fast, reliable analytical methods has made possible the qualitative and quantitative assessment of technetium species generated by the radiolabeling process. Labeling stability is determined by competition of the 99m Tc-antibody bond with three ligands, Chelex 100 (a metal chelate-type resin), free DTPA solution and 1% HSA solution. Very good 99m Tc-antibody stability is obtained with activated IgG (IgGa) and Fab' fragment, which makes these substances possible candidates for immunoscintigraphy use. (author)

Making microenvironments: A look into incorporating macromolecular crowding into in vitro experiments, to generate biomimetic microenvironments which are capable of directing cell function for tissue engineering applications.

Science.gov (United States)

Benny, Paula; Raghunath, Michael

2017-01-01

Biomimetic microenvironments are key components to successful cell culture and tissue engineering in vitro. One of the most accurate biomimetic microenvironments is that made by the cells themselves. Cell-made microenvironments are most similar to the in vivo state as they are cell-specific and produced by the actual cells which reside in that specific microenvironment. However, cell-made microenvironments have been challenging to re-create in vitro due to the lack of extracellular matrix composition, volume and complexity which are required. By applying macromolecular crowding to current cell culture protocols, cell-made microenvironments, or cell-derived matrices, can be generated at significant rates in vitro. In this review, we will examine the causes and effects of macromolecular crowding and how it has been applied in several in vitro systems including tissue engineering.

Designing food delivery systems: challenges related to the in vitro methods employed to determine the fate of bioactives in the gut.

Science.gov (United States)

Arranz, Elena; Corredig, Milena; Guri, Anilda

2016-08-10

An in depth understanding of the underpinning mechanisms that relate to food disruption and processing in the gastrointestinal tract is necessary to achieve optimal intake of nutrients and their bioefficacy. Although in vivo trials can provide insights on physiological responses of nutrients, in vitro assays are often applied as tools to understand specific mechanisms, or as prescreening methods to determine the factors associated with the uptake of food components in the gastrointestinal tract. In vitro assays are also often utilized to design novel or improved food delivery systems. In this review the available approaches to study delivery and uptake of food bioactives and the associated challenges are discussed. For an in depth understanding of food processing in the gastrointestinal tract, it is necessary to apply multidisciplinary methodologies, at the interface between materials science, chemistry, physics and biology.

Wireless Sensor Network Security Enhancement Using Directional Antennas: State of the Art and Research Challenges.

Science.gov (United States)

Curiac, Daniel-Ioan

2016-04-07

Being often deployed in remote or hostile environments, wireless sensor networks are vulnerable to various types of security attacks. A possible solution to reduce the security risks is to use directional antennas instead of omnidirectional ones or in conjunction with them. Due to their increased complexity, higher costs and larger sizes, directional antennas are not traditionally used in wireless sensor networks, but recent technology trends may support this method. This paper surveys existing state of the art approaches in the field, offering a broad perspective of the future use of directional antennas in mitigating security risks, together with new challenges and open research issues.

In Vitro Maturation (IVM of Human Oocytes: Promising Potential, Challenges and Chances for Improvement

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Roza silvia

2015-05-01

Full Text Available AbstractIn Vitro Maturation (IVM of human oocytes is an innovation in Assisted Reproductive Technology (ART. It is believed more patient-friendly than conventional In Vitro Fertilization (IVF method. It is a simple protocol that needs only less injection of ovarian stimulation for the patients and fewer blood sample and ultrasound scans, so this technique may become more favorable. Patients are also prevented from higher cost treatments and quite long control in the hospital. However, there are some problems to be addressed, such as how to improve the success rate, how to assure the safety and to avoid the health risk for the offsprings. Modification in IVM medium and optimizing the IVM protocols have increased the results in some studies. However, further investigation related to all aspects influencing the human oocyte maturation in vitro is still needed to make it enable to be a routine practice in ART centers for a defined group.Kata kunci: in vitro maturation, human oocyte, in vitro fertilization, assisted reproductive technology AbstrakMaturasi oosit in vitro atau In Vitro Maturation (IVM terhadap oosit manusia merupakan suatu inovasi dalam Teknologi Reproduksi Berbantu (TRB. Teknik ini dianggap lebih nyaman bagi pasien dibandingkan dengan metode Fertilisasi In Vitro (FIV konvensional. Metode IVM ini sederhana dan hanya membutuhkan lebih sedikit penyuntikan obat stimulasi ovarium ke pasien serta lebih sedikit pemeriksaan darah dan ultrasonografi, sehingga memungkinkan untuk menjadi suatu pilihan yang disukai oleh pasien. Pasien juga bisa terhindar dari biaya terapi yang lebih mahal serta waktu kontrol yang lama di rumah sakit. Namun demikian, terdapat beberapa masalah yang perlu ditangani terkait metode ini, seperti bagaimana meningkatkan angka keberhasilan serta memastikan keamanan dan mencegah resiko kesehatan pada anak yang akan dilahirkan. Modifikasi pada medium IVM serta pengoptimalan protokol IVM telah meningkatkan hasil pada

Direct Cardiac Reprogramming: Advances in Cardiac Regeneration

Directory of Open Access Journals (Sweden)

Olivia Chen

2015-01-01

Full Text Available Heart disease is one of the lead causes of death worldwide. Many forms of heart disease, including myocardial infarction and pressure-loading cardiomyopathies, result in irreversible cardiomyocyte death. Activated fibroblasts respond to cardiac injury by forming scar tissue, but ultimately this response fails to restore cardiac function. Unfortunately, the human heart has little regenerative ability and long-term outcomes following acute coronary events often include chronic and end-stage heart failure. Building upon years of research aimed at restoring functional cardiomyocytes, recent advances have been made in the direct reprogramming of fibroblasts toward a cardiomyocyte cell fate both in vitro and in vivo. Several experiments show functional improvements in mouse models of myocardial infarction following in situ generation of cardiomyocyte-like cells from endogenous fibroblasts. Though many of these studies are in an early stage, this nascent technology holds promise for future applications in regenerative medicine. In this review, we discuss the history, progress, methods, challenges, and future directions of direct cardiac reprogramming.

Challenges in Discerning Atmospheric Composition in Directly Imaged Planets

Science.gov (United States)

Marley, Mark S.

2017-01-01

One of the justifications motivating efforts to detect and characterize young extrasolar giant planets has been to measure atmospheric composition for comparison with that of the primary star. If the enhancement of heavy elements in the atmospheres of extrasolar giant planets, like it is for their solar system analogs, is inversely proportional to mass, then it is likely that these worlds formed by core accretion. However in practice it has been very difficult to constrain metallicity because of the complex effect of clouds. Cloud opacity varies both vertically and, in some cases, horizontally through the atmosphere. Particle size and composition, both of which impact opacity, are difficult challenges both for forward modeling and retrieval studies. In my presentation I will discuss systematic efforts to improve cloud studies to enable more reliable determinations of atmospheric composition. These efforts are relevant both to discerning composition of directly imaged young planets from ground based telescopes and future space based missions, such as WFIRST and LUVOIR.

In vivo reprogramming for heart regeneration: A glance at efficiency, environmental impacts, challenges and future directions.

Science.gov (United States)

Ebrahimi, Behnam

2017-07-01

Replacing dying or diseased cells of a tissue with new ones that are converted from patient's own cells is an attractive strategy in regenerative medicine. In vivo reprogramming is a novel strategy that can circumvent the hurdles of autologous/allogeneic cell injection therapies. Interestingly, studies have demonstrated that direct injection of cardiac transcription factors or specific miRNAs into the infarct border zone of murine hearts following myocardial infarction converts resident cardiac fibroblasts into functional cardiomyocytes. Moreover, in vivo cardiac reprogramming not only drives cardiac tissue regeneration, but also improves cardiac function and survival rate after myocardial infarction. Thanks to the influence of cardiac microenvironment and the same developmental origin, cardiac fibroblasts seem to be more amenable to reprogramming toward cardiomyocyte fate than other cell sources (e.g. skin fibroblasts). Thus, reprogramming of cardiac fibroblasts to functional induced cardiomyocytes in the cardiac environment holds great promises for induced regeneration and potential clinical purposes. Application of small molecules in future studies may represent a major advancement in this arena and pharmacological reprogramming would convey reprogramming technology to the translational medicine paradigm. This study reviews accomplishments in the field of in vitro and in vivo mouse cardiac reprogramming and then deals with strategies for the enhancement of the efficiency and quality of the process. Furthermore, it discusses challenges ahead and provides suggestions for future research. Human cardiac reprogramming is also addressed as a foundation for possible application of in vivo cardiac reprogramming for human heart regeneration in the future. Copyright © 2017 Elsevier Ltd. All rights reserved.

In vitro rapid multiplication of Stevia rebaudiana: an important natural sweetener herb

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RAVINDRA ADE

2012-11-01

Full Text Available Deshmukh S, Ade R. 2012. In vitro rapid multiplication of Stevia rebaudiana: an important natural sweetener herb. Nusantara Bioscience 4: 105-108. Stevia rebaudiana Bertoni, belonging to family Asteraceae and natural sweet plant, but due to poor seed viability, fertility and vigor, Stevia cultivation is a challenging task. In the present study in vitro rapid multiplication method was established for Stevia rebaudiana by inoculating explants on M.S. medium, supplemented with different combination of phytoharmone. The maximum number of shoots (18.3±0.8 was obtained on M.S. medium supplemented with BAP + KIN (1.5 + 0.5 mg/L. The highest rooting percentage (95.25 was observed with (IAA 0.1 mg/L. The rooted plants were successfully established firstly in soil with coco peat (1:1 and then directly in ordinary soil.

A BMP7 Variant Inhibits Tumor Angiogenesis In Vitro and In Vivo through Direct Modulation of Endothelial Cell Biology.

Directory of Open Access Journals (Sweden)

Courtney M Tate

Full Text Available Bone morphogenetic proteins (BMPs, members of the TGF-β superfamily, have numerous biological activities including control of growth, differentiation, and vascular development. Using an in vitro co-culture endothelial cord formation assay, we investigated the role of a BMP7 variant (BMP7v in VEGF, bFGF, and tumor-driven angiogenesis. BMP7v treatment led to disruption of neo-endothelial cord formation and regression of existing VEGF and bFGF cords in vitro. Using a series of tumor cell models capable of driving angiogenesis in vitro, BMP7v treatment completely blocked cord formation. Pre-treatment of endothelial cells with BMP7v significantly reduced their cord forming ability, indicating a direct effect on endothelial cell function. BMP7v activated the canonical SMAD signaling pathway in endothelial cells but targeted gene knockdown using shRNA directed against SMAD4 suggests this pathway is not required to mediate the anti-angiogenic effect. In contrast to SMAD activation, BMP7v selectively decreased ERK and AKT activation, significantly decreased endothelial cell migration and down-regulated expression of critical RTKs involved in VEGF and FGF angiogenic signaling, VEGFR2 and FGFR1 respectively. Importantly, in an in vivo angiogenic plug assay that serves as a measurement of angiogenesis, BMP7v significantly decreased hemoglobin content indicating inhibition of neoangiogenesis. In addition, BMP7v significantly decreased angiogenesis in glioblastoma stem-like cell (GSLC Matrigel plugs and significantly impaired in vivo growth of a GSLC xenograft with a concomitant reduction in microvessel density. These data support BMP7v as a potent anti-angiogenic molecule that is effective in the context of tumor angiogenesis.

A BMP7 Variant Inhibits Tumor Angiogenesis In Vitro and In Vivo through Direct Modulation of Endothelial Cell Biology.

Science.gov (United States)

Tate, Courtney M; Mc Entire, Jacquelyn; Pallini, Roberto; Vakana, Eliza; Wyss, Lisa; Blosser, Wayne; Ricci-Vitiani, Lucia; D'Alessandris, Quintino Giorgio; Morgante, Liliana; Giannetti, Stefano; Larocca, Luigi Maria; Todaro, Matilde; Benfante, Antonina; Colorito, Maria Luisa; Stassi, Giorgio; De Maria, Ruggero; Rowlinson, Scott; Stancato, Louis

2015-01-01

Bone morphogenetic proteins (BMPs), members of the TGF-β superfamily, have numerous biological activities including control of growth, differentiation, and vascular development. Using an in vitro co-culture endothelial cord formation assay, we investigated the role of a BMP7 variant (BMP7v) in VEGF, bFGF, and tumor-driven angiogenesis. BMP7v treatment led to disruption of neo-endothelial cord formation and regression of existing VEGF and bFGF cords in vitro. Using a series of tumor cell models capable of driving angiogenesis in vitro, BMP7v treatment completely blocked cord formation. Pre-treatment of endothelial cells with BMP7v significantly reduced their cord forming ability, indicating a direct effect on endothelial cell function. BMP7v activated the canonical SMAD signaling pathway in endothelial cells but targeted gene knockdown using shRNA directed against SMAD4 suggests this pathway is not required to mediate the anti-angiogenic effect. In contrast to SMAD activation, BMP7v selectively decreased ERK and AKT activation, significantly decreased endothelial cell migration and down-regulated expression of critical RTKs involved in VEGF and FGF angiogenic signaling, VEGFR2 and FGFR1 respectively. Importantly, in an in vivo angiogenic plug assay that serves as a measurement of angiogenesis, BMP7v significantly decreased hemoglobin content indicating inhibition of neoangiogenesis. In addition, BMP7v significantly decreased angiogenesis in glioblastoma stem-like cell (GSLC) Matrigel plugs and significantly impaired in vivo growth of a GSLC xenograft with a concomitant reduction in microvessel density. These data support BMP7v as a potent anti-angiogenic molecule that is effective in the context of tumor angiogenesis.

What is in vitro meat? Food phreaking

OpenAIRE

Stephens, NS

2015-01-01

This book gathers the ideas and opinions of a number of scientists and other experts around the topic of in vitro meat. The authors in this publication range from being in vitro meat’s developers and most vocal supporters, to some adamant opposers. Collectively, these essays present a diversity of perspectives, and illustrate the challenge of pinning down an emerging technology.

Modelling water use in global hydrological models: review, challenges and directions

Science.gov (United States)

Bierkens, M. F.; de Graaf, I.; Wada, Y.; Wanders, N.; Van Beek, L. P.

2017-12-01

During the late 1980s and early 1990s, awareness of the shortage of global water resources lead to the first detailed global water resources assessments using regional statistics of water use and observations of meteorological and hydrological variables. Shortly thereafter, the first macroscale hydrological models (MHM) appeared. In these models, blue water (i.e., surface water and renewable groundwater) availability was calculated by accumulating runoff over a stream network and comparing it with population densities or with estimated water demand for agriculture, industry and households. In this talk we review the evolution of human impact modelling in global land models with a focus on global water resources, touching upon developments of the last 15 years: i.e. calculating human water scarcity; estimating groundwater depletion; adding dams and reservoirs; fully integrating water use (demand, withdrawal, consumption, return flow) in the hydrology; simulating the effects of land use change. We show example studies for each of these steps. We identify We identify major challenges that hamper the further development of integrated water resources modelling. Examples of these are: 1) simulating reservoir operations; 2) including local infrastructure and redistribution; 3) using the correct allocations rules; 4) projecting future water demand and water use. For each of these challenges we signify promising directions for further research.

De Novo Human Cardiac Myocytes for Medical Research: Promises and Challenges

Directory of Open Access Journals (Sweden)

Veronique Hamel

2017-01-01

Full Text Available The advent of cellular reprogramming technology has revolutionized biomedical research. De novo human cardiac myocytes can now be obtained from direct reprogramming of somatic cells (such as fibroblasts, from induced pluripotent stem cells (iPSCs, which are reprogrammed from somatic cells, and from human embryonic stem cells (hESCs. Such de novo human cardiac myocytes hold great promise for in vitro disease modeling and drug screening and in vivo cell therapy of heart disease. Here, we review the technique advancements for generating de novo human cardiac myocytes. We also discuss several challenges for the use of such cells in research and regenerative medicine, such as the immature phenotype and heterogeneity of de novo cardiac myocytes obtained with existing protocols. We focus on the recent advancements in addressing such challenges.

The challenges of transportation/traffic statistics in Japan and directions for the future

Directory of Open Access Journals (Sweden)

Shigeru Kawasaki

2015-07-01

Full Text Available In order to respond to new challenges in transportation and traffic problems, it is essential to enhance statistics in this field that provides the basis for policy researches. Many of the statistics in this field in Japan consist of “official statistics” created by the government. This paper gives a review of the current status of transportation and traffic statistics (hereinafter called “transportation statistics” in short in Japan. Furthermore, the paper discusses challenges in such statistics in the new environment and the direction that statistics that should take in the future. For Japan’s transportation statistics to play vital roles in more sophisticated analyses, it is necessary to improve the environment that facilitates the use of microdata for analysis. It is also necessary to establish an environment where big data can be more easily used for compilation of official statistics and performing policy researches. To achieve this end, close cooperation among the government, academia, and businesses will be essential.

Application of In Vitro Transposon Mutagenesis to Erythromycin Strain Improvement in Saccharopolyspora erythraea.

Science.gov (United States)

Weber, J Mark; Reeves, Andrew; Cernota, William H; Wesley, Roy K

2017-01-01

Transposon mutagenesis is an invaluable technique in molecular biology for the creation of random mutations that can be easily identified and mapped. However, in the field of microbial strain improvement, transposon mutagenesis has scarcely been used; instead, chemical and physical mutagenic methods have been traditionally favored. Transposons have the advantage of creating single mutations in the genome, making phenotype to genotype assignments less challenging than with traditional mutagens which commonly create multiple mutations in the genome. The site of a transposon mutation can also be readily mapped using DNA sequencing primer sites engineered into the transposon termini. In this chapter an in vitro method for transposon mutagenesis of Saccharopolyspora erythraea is presented. Since in vivo transposon tools are not available for most actinomycetes including S. erythraea, an in vitro method was developed. The in vitro method involves a significant investment in time and effort to create the mutants, but once the mutants are made and screened, a large number of highly relevant mutations of direct interest to erythromycin production can be found.

Implementing Indigenous Education Policy Directives in Ontario Public Schools: Experiences, Challenges and Successful Practices

Directory of Open Access Journals (Sweden)

Emily Milne

2017-08-01

Full Text Available The Ontario Ministry of Education has declared a commitment to Indigenous student success and has advanced a policy framework that articulates inclusion of Indigenous content in schooling curriculum (Ontario Ministry of Education, 2007. What are the perceptions among educators and parents regarding the implementation of policy directives, and what is seen to encourage or limit meaningful implementation? To answer these questions, this article draws on interviews with 100 Indigenous (mainly Haudenosaunee, Anishinaabe, and Métis and non-Indigenous parents and educators from Ontario Canada. Policy directives are seen to benefit Indigenous and non-Indigenous students. Interviews also reveal challenges to implementing Indigenous curricular policy, such as unawareness and intimidation among non-Indigenous educators regarding how to teach material. Policy implications are considered.

Stable subcutaneous cartilage regeneration of bone marrow stromal cells directed by chondrocyte sheet.

Science.gov (United States)

Li, Dan; Zhu, Lian; Liu, Yu; Yin, Zongqi; Liu, Yi; Liu, Fangjun; He, Aijuan; Feng, Shaoqing; Zhang, Yixin; Zhang, Zhiyong; Zhang, Wenjie; Liu, Wei; Cao, Yilin; Zhou, Guangdong

2017-05-01

In vivo niche plays an important role in regulating differentiation fate of stem cells. Due to lack of proper chondrogenic niche, stable cartilage regeneration of bone marrow stromal cells (BMSCs) in subcutaneous environments is always a great challenge. This study explored the feasibility that chondrocyte sheet created chondrogenic niche retained chondrogenic phenotype of BMSC engineered cartilage (BEC) in subcutaneous environments. Porcine BMSCs were seeded into biodegradable scaffolds followed by 4weeks of chondrogenic induction in vitro to form BEC, which were wrapped with chondrocyte sheets (Sheet group), acellular small intestinal submucosa (SIS, SIS group), or nothing (Blank group) respectively and then implanted subcutaneously into nude mice to trace the maintenance of chondrogenic phenotype. The results showed that all the constructs in Sheet group displayed typical cartilaginous features with abundant lacunae and cartilage specific matrices deposition. These samples became more mature with prolonged in vivo implantation, and few signs of ossification were observed at all time points except for one sample that had not been wrapped completely. Cell labeling results in Sheet group further revealed that the implanted BEC directly participated in cartilage formation. Samples in both SIS and Blank groups mainly showed ossified tissue at all time points with partial fibrogenesis in a few samples. These results suggested that chondrocyte sheet could create a chondrogenic niche for retaining chondrogenic phenotype of BEC in subcutaneous environment and thus provide a novel research model for stable ectopic cartilage regeneration based on stem cells. In vivo niche plays an important role in directing differentiation fate of stem cells. Due to lack of proper chondrogenic niche, stable cartilage regeneration of bone marrow stromal cells (BMSCs) in subcutaneous environments is always a great challenge. The current study demonstrated that chondrocyte sheet generated by

21 CFR 201.119 - In vitro diagnostic products.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 4 2010-04-01 2010-04-01 false In vitro diagnostic products. 201.119 Section 201...) DRUGS: GENERAL LABELING Exemptions From Adequate Directions for Use § 201.119 In vitro diagnostic products. (a) “In vitro diagnostic products” are those reagents, instruments and systems intended for use...

21 CFR 801.119 - In vitro diagnostic products.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false In vitro diagnostic products. 801.119 Section 801...) MEDICAL DEVICES LABELING Exemptions From Adequate Directions for Use § 801.119 In vitro diagnostic products. A product intended for use in the diagnosis of disease and which is an in vitro diagnostic...

Determining the direction of tooth grinding: an in vitro study

NARCIS (Netherlands)

ten Berge, F.; te Poel, J.; Ranjitkar, S.; Kaidonis, J.A.; Lobbezoo, F.; Hughes, T.E.; Townsend, G.C.

2012-01-01

The analysis of microwear patterns, including scratch types and widths, has enabled reconstruction of the dietary habits and lifestyles of prehistoric and modern humans. The aim of this in vitro study was to determine whether an assessment of microwear features of experimental scratches placed on

In vitro methods for hazard assessment of industrial chemicals – opportunities and challenges

Directory of Open Access Journals (Sweden)

Chin Lin eWong

2015-05-01

Full Text Available Allergic contact dermatitis (ACD is a delayed-type hypersensitivity immune reaction mediated by T-lymphocytes as a result of repeated exposure of an allergen primarily on skin. ACD accounts for up to 95% of occupational skin diseases (OSDs, with epoxy resins implicated as one of the most common causes of ACD. Efficient high-throughput in vitro screening for accurate identification of compounds and materials that may pose hazardous risks in the workplace is crucial. At present, the murine local lymph node assay (LLNA is the ‘method of choice’ for predicting the sensitizing potency of contact allergens. As the 3Rs principles of reduction, refinement and replacement in animal testing has gained political and economic momentum, several in vitro screening methods have been developed for identifying potential contact allergens. To date, these latter methods have been utilized primarily to assess the skin sensitizing potential of the chemical components of cosmetic products with scant research attention as to the applicability of these methods to industrial chemicals, particularly epoxy resins. Herein we review the currently utilized in vitro methods and identify the knowledge gaps with regard to assessing the generalizability of in vitro screening methods for assessing the skin sensitizing potential of industrial chemicals.

Assessment approaches in massive open online courses: Possibilities, challenges and future directions

Science.gov (United States)

Xiong, Yao; Suen, Hoi K.

2018-03-01

The development of massive open online courses (MOOCs) has launched an era of large-scale interactive participation in education. While massive open enrolment and the advances of learning technology are creating exciting potentials for lifelong learning in formal and informal ways, the implementation of efficient and effective assessment is still problematic. To ensure that genuine learning occurs, both assessments for learning (formative assessments), which evaluate students' current progress, and assessments of learning (summative assessments), which record students' cumulative progress, are needed. Providers' more recent shift towards the granting of certificates and digital badges for course accomplishments also indicates the need for proper, secure and accurate assessment results to ensure accountability. This article examines possible assessment approaches that fit open online education from formative and summative assessment perspectives. The authors discuss the importance of, and challenges to, implementing assessments of MOOC learners' progress for both purposes. Various formative and summative assessment approaches are then identified. The authors examine and analyse their respective advantages and disadvantages. They conclude that peer assessment is quite possibly the only universally applicable approach in massive open online education. They discuss the promises, practical and technical challenges, current developments in and recommendations for implementing peer assessment. They also suggest some possible future research directions.

Understanding the Self-Directed Online Learning Preferences, Goals, Achievements, and Challenges of MIT OpenCourseWare Subscribers

Science.gov (United States)

Bonk, Curtis J.; Lee, Mimi Miyoung; Kou, Xiaojing; Xu, Shuya; Sheu, Feng-Ru

2015-01-01

This research targeted the learning preferences, goals and motivations, achievements, challenges, and possibilities for life change of self-directed online learners who subscribed to the monthly OpenCourseWare (OCW) e-newsletter from MIT. Data collection included a 25-item survey of 1,429 newsletter subscribers; 613 of whom also completed an…

Differential effects of allergen challenge on large and small airway reactivity in mice.

Directory of Open Access Journals (Sweden)

Chantal Donovan

Full Text Available The relative contributions of large and small airways to hyperresponsiveness in asthma have yet to be fully assessed. This study used a mouse model of chronic allergic airways disease to induce inflammation and remodelling and determine whether in vivo hyperresponsiveness to methacholine is consistent with in vitro reactivity of trachea and small airways. Balb/C mice were sensitised (days 0, 14 and challenged (3 times/week, 6 weeks with ovalbumin. Airway reactivity was compared with saline-challenged controls in vivo assessing whole lung resistance, and in vitro measuring the force of tracheal contraction and the magnitude/rate of small airway narrowing within lung slices. Increased airway inflammation, epithelial remodelling and fibrosis were evident following allergen challenge. In vivo hyperresponsiveness to methacholine was maintained in isolated trachea. In contrast, methacholine induced slower narrowing, with reduced potency in small airways compared to controls. In vitro incubation with IL-1/TNFα did not alter reactivity. The hyporesponsiveness to methacholine in small airways within lung slices following chronic ovalbumin challenge was unexpected, given hyperresponsiveness to the same agonist both in vivo and in vitro in tracheal preparations. This finding may reflect the altered interactions of small airways with surrounding parenchymal tissue after allergen challenge to oppose airway narrowing and closure.

Public Service Motivation Research : Achievements, Challenges, and Future Directions

NARCIS (Netherlands)

Perry, James L.; Vandenabeele, Wouter

2015-01-01

This article takes stock of public service motivation research to identify achievements, challenges, and an agenda for research to build on progress made since 1990. After enumerating achievements and challenges, the authors take stock of progress on extant proposals to strengthen research. In

Rethinking the globalisation of problem-based learning: how culture challenges self-directed learning.

Science.gov (United States)

Frambach, Janneke M; Driessen, Erik W; Chan, Li-Chong; van der Vleuten, Cees P M

2012-08-01

Medical schools worldwide are increasingly switching to student-centred methods such as problem-based learning (PBL) to foster lifelong self-directed learning (SDL). The cross-cultural applicability of these methods has been questioned because of their Western origins and because education contexts and learning approaches differ across cultures. This study evaluated PBL's cross-cultural applicability by investigating how it is applied in three medical schools in regions with different cultures in, respectively, East Asia, the Middle East and Western Europe. Specifically, it investigated how students' cultural backgrounds impact on SDL in PBL and how this impact affects students. A qualitative, cross-cultural, comparative case study was conducted in three medical schools. Data were collected through 88 semi-structured, in-depth interviews with Year 1 and 3 students, tutors and key persons involved in PBL, 32 observations of Year 1 and 3 PBL tutorials, document analysis, and contextual information. The data were thematically analysed using the template analysis method. Comparisons were made among the three medical schools and between Year 1 and 3 students across and within the schools. The cultural factors of uncertainty and tradition posed a challenge to Middle Eastern students' SDL. Hierarchy posed a challenge to Asian students and achievement impacted on both sets of non-Western students. These factors were less applicable to European students, although the latter did experience some challenges. Several contextual factors inhibited or enhanced SDL across the cases. As students grew used to PBL, SDL skills increased across the cases, albeit to different degrees. Although cultural factors can pose a challenge to the application of PBL in non-Western settings, it appears that PBL can be applied in different cultural contexts. However, its globalisation does not postulate uniform processes and outcomes, and culturally sensitive alternatives might be developed. �

Fall Prediction and Prevention Systems: Recent Trends, Challenges, and Future Research Directions

Directory of Open Access Journals (Sweden)

Ramesh Rajagopalan

2017-11-01

Full Text Available Fall prediction is a multifaceted problem that involves complex interactions between physiological, behavioral, and environmental factors. Existing fall detection and prediction systems mainly focus on physiological factors such as gait, vision, and cognition, and do not address the multifactorial nature of falls. In addition, these systems lack efficient user interfaces and feedback for preventing future falls. Recent advances in internet of things (IoT and mobile technologies offer ample opportunities for integrating contextual information about patient behavior and environment along with physiological health data for predicting falls. This article reviews the state-of-the-art in fall detection and prediction systems. It also describes the challenges, limitations, and future directions in the design and implementation of effective fall prediction and prevention systems.

Fall Prediction and Prevention Systems: Recent Trends, Challenges, and Future Research Directions.

Science.gov (United States)

Rajagopalan, Ramesh; Litvan, Irene; Jung, Tzyy-Ping

2017-11-01

Fall prediction is a multifaceted problem that involves complex interactions between physiological, behavioral, and environmental factors. Existing fall detection and prediction systems mainly focus on physiological factors such as gait, vision, and cognition, and do not address the multifactorial nature of falls. In addition, these systems lack efficient user interfaces and feedback for preventing future falls. Recent advances in internet of things (IoT) and mobile technologies offer ample opportunities for integrating contextual information about patient behavior and environment along with physiological health data for predicting falls. This article reviews the state-of-the-art in fall detection and prediction systems. It also describes the challenges, limitations, and future directions in the design and implementation of effective fall prediction and prevention systems.

Retroviral DNA Integration Directed by HIV Integration Protein in Vitro

Science.gov (United States)

Bushman, Frederic D.; Fujiwara, Tamio; Craigie, Robert

1990-09-01

Efficient retroviral growth requires integration of a DNA copy of the viral RNA genome into a chromosome of the host. As a first step in analyzing the mechanism of integration of human immunodeficiency virus (HIV) DNA, a cell-free system was established that models the integration reaction. The in vitro system depends on the HIV integration (IN) protein, which was partially purified from insect cells engineered to express IN protein in large quantities. Integration was detected in a biological assay that scores the insertion of a linear DNA containing HIV terminal sequences into a λ DNA target. Some integration products generated in this assay contained five-base pair duplications of the target DNA at the recombination junctions, a characteristic of HIV integration in vivo; the remaining products contained aberrant junctional sequences that may have been produced in a variation of the normal reaction. These results indicate that HIV IN protein is the only viral protein required to insert model HIV DNA sequences into a target DNA in vitro.

In vitro cumulative gas production techniques: history, methodological considerations and challenges

NARCIS (Netherlands)

Rymer, C.; Huntington, J.A.; Williams, B.A.; Givens, D.I.

2005-01-01

Methodology used to measure in vitro gas production is reviewed to determine impacts of sources of variation on resultant gas production profiles (GPP). Current methods include measurement of gas production at constant pressure (e.g., use of gas tight syringes), a system that is inexpensive, but may

Accounting for data variability, a key factor in in vivo/in vitro relationships: application to the skin sensitization potency (in vivo LLNA versus in vitro DPRA) example.

Science.gov (United States)

Dimitrov, S; Detroyer, A; Piroird, C; Gomes, C; Eilstein, J; Pauloin, T; Kuseva, C; Ivanova, H; Popova, I; Karakolev, Y; Ringeissen, S; Mekenyan, O

2016-12-01

When searching for alternative methods to animal testing, confidently rescaling an in vitro result to the corresponding in vivo classification is still a challenging problem. Although one of the most important factors affecting good correlation is sample characteristics, they are very rarely integrated into correlation studies. Usually, in these studies, it is implicitly assumed that both compared values are error-free numbers, which they are not. In this work, we propose a general methodology to analyze and integrate data variability and thus confidence estimation when rescaling from one test to another. The methodology is demonstrated through the case study of rescaling the in vitro Direct Peptide Reactivity Assay (DPRA) reactivity to the in vivo Local Lymph Node Assay (LLNA) skin sensitization potency classifications. In a first step, a comprehensive statistical analysis evaluating the reliability and variability of LLNA and DPRA as such was done. These results allowed us to link the concept of gray zones and confidence probability, which in turn represents a new perspective for a more precise knowledge of the classification of chemicals within their in vivo OR in vitro test. Next, the novelty and practical value of our methodology introducing variability into the threshold optimization between the in vitro AND in vivo test resides in the fact that it attributes a confidence probability to the predicted classification. The methodology, classification and screening approach presented in this study are not restricted to skin sensitization only. They could be helpful also for fate, toxicity and health hazard assessment where plenty of in vitro and in chemico assays and/or QSARs models are available. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Challenges and future direction of molecular research in air pollution-related lung cancers.

Science.gov (United States)

Shahadin, Maizatul Syafinaz; Ab Mutalib, Nurul Syakima; Latif, Mohd Talib; Greene, Catherine M; Hassan, Tidi

2018-04-01

Hazardous air pollutants or chemical release into the environment by a variety of natural and/or anthropogenic activities may give adverse effects to human health. Air pollutants such as sulphur dioxide (SO2), nitrogen oxides (NOx), carbon monoxide (CO), heavy metals and particulate matter (PM) affect number of different human organs, especially the respiratory system. The International Agency for Research on Cancer (IARC) reported that ambient air pollution is a cause of lung cancer. Recently, the agency has classified outdoor air pollution as well as PM air pollution as Group 1 carcinogens. In addition, several epidemiological studies have shown a positive association between air pollutants to lung cancer risks and mortality. However, there are only a few studies examining the molecular effects of air pollution exposure specifically in lung cancer due to multiple challenges to mimic air pollution exposure in basic experimentation. Another major issue is the lack of adequate adjustments for exposure misclassification as air pollution may differ temporo-spatially and socioeconomically. Thus, the purpose of this paper is to review the current molecular understanding of air pollution-related lung cancer and potential future direction in this challenging yet important research field. Copyright © 2018 Elsevier B.V. All rights reserved.

Direct effects of casein phosphopeptides on growth and differentiation of in vitro cultured osteoblastic cells (MC3T3-E1).

Science.gov (United States)

Tulipano, Giovanni; Bulgari, Omar; Chessa, Stefania; Nardone, Alessandro; Cocchi, Daniela; Caroli, Anna

2010-02-25

Casein phosphopeptides (CPPs) obtained by enzymatic hydrolysis in vitro of caseins, have been shown to enhance calcium solubility and to increase the calcification of embryonic rat bones in their diaphyseal area. Little is known about the direct effects of CPPs on cultured osteoblastic cells. Calcium in the microenvironment surrounding bone cells is not only important for the mineralization of the extracellular matrix, but it is believed to provide preosteblasts with a signal that modulates their proliferation and differentiation. The aim of the present study was to investigate the direct effects of four selected casein phosphopeptides on osteoblastic cell (MC3T3-E1 cells) viability and differentiation. The selected peptides have been obtained by chemical synthesis and differed in the number of phosphorylated sites and in the amino acid spacing out two phosphorylated sites, in order to further characterize the relationship between structure and function. The results obtained in this work demonstrated that CPPs may directly affect osteoblast-like cell growth, calcium uptake and ultimately calcium deposition in the extracellular matrix. The effects exerted by distinct CPPs on osteogenesis in vitro can be either stimulatory or inhibitory. Differential short amino acid sequences in their molecules, like the -SpEE- and the -SpTSpEE-motifs, are likely the molecular determinants for their biological activities on osteoblastic cells. Moreover, two genetic variants of CPPs showing one amino acid change in their sequence may profoundly differ in their biological activities. Finally, our data may also suggest important clues about the role of intrinsic phosphorylated peptides derived from endogenous phosphorylated proteins in bone metabolism, apart from extrinsic CPPs. Copyright 2009 Elsevier B.V. All rights reserved.

Isolation and characterization of monoclonal antibodies directed against two subunits of rabbit poxvirus-associated, DNA-directed RNA polymerase.

OpenAIRE

Morrison, D K; Carter, J K; Moyer, R W

1985-01-01

A library of monoclonal antibodies directed against individual proteins of the rabbit poxvirus (RPV) virion within a complex immunogenic mixture has been generated through the use of in vivo and in vitro immunization regimens. The relative efficacies of the two procedures were compared. Based on immunoblot analysis, the in vitro immunization regimen led both to a wider variety of monoclonal antibodies to different proteins and to a larger number of antibodies directed against proteins of high...

The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro

Directory of Open Access Journals (Sweden)

Gronenborn Angela M

2010-05-01

Full Text Available Abstract Background Lemay et al recently reported that the RNA binding protein HuR directly interacts with the ribonuclease H (RNase H domain of HIV-1 reverse transcriptase (RT and influences the efficiency of viral reverse transcription (Lemay et al., 2008, Retrovirology 5:47. HuR is a member of the embryonic lethal abnormal vision protein family and contains 3 RNA recognition motifs (RRMs that bind AU-rich elements (AREs. To define the structural determinants of the HuR-RT interaction and to elucidate the mechanism(s by which HuR influences HIV-1 reverse transcription activity in vitro, we cloned and purified full-length HuR as well as three additional protein constructs that contained the N-terminal and internal RRMs, the internal and C-terminal RRMs, or the C-terminal RRM only. Results All four HuR proteins were purified and characterized by biophysical methods. They are well structured and exist as monomers in solution. No direct protein-protein interaction between HuR and HIV-1 RT was detected using NMR titrations with 15N labeled HuR variants or the 15N labeled RNase H domain of HIV-1 RT. Furthermore, HuR did not significantly affect the kinetics of HIV-1 reverse transcription in vitro, even on RNA templates that contain AREs. Conclusions Our results suggest that HuR does not impact HIV-1 replication through a direct protein-protein interaction with the viral RT.

Untargeted Metabolomics Strategies—Challenges and Emerging Directions

Science.gov (United States)

Schrimpe-Rutledge, Alexandra C.; Codreanu, Simona G.; Sherrod, Stacy D.; McLean, John A.

2016-12-01

Metabolites are building blocks of cellular function. These species are involved in enzyme-catalyzed chemical reactions and are essential for cellular function. Upstream biological disruptions result in a series of metabolomic changes and, as such, the metabolome holds a wealth of information that is thought to be most predictive of phenotype. Uncovering this knowledge is a work in progress. The field of metabolomics is still maturing; the community has leveraged proteomics experience when applicable and developed a range of sample preparation and instrument methodology along with myriad data processing and analysis approaches. Research focuses have now shifted toward a fundamental understanding of the biology responsible for metabolomic changes. There are several types of metabolomics experiments including both targeted and untargeted analyses. While untargeted, hypothesis generating workflows exhibit many valuable attributes, challenges inherent to the approach remain. This Critical Insight comments on these challenges, focusing on the identification process of LC-MS-based untargeted metabolomics studies—specifically in mammalian systems. Biological interpretation of metabolomics data hinges on the ability to accurately identify metabolites. The range of confidence associated with identifications that is often overlooked is reviewed, and opportunities for advancing the metabolomics field are described.

A different challenge: the directional drilled crossing for the Yacuiba - Rio Grande Gas Line Project - GASYRG

Energy Technology Data Exchange (ETDEWEB)

Green, Wayne; Garcia, Francisco [Bolinter Ltda., Santa Cruz (Bolivia); Montano, Ruben [Transierra, Santa Cruz (Bolivia)

2003-07-01

The Rio Grande River's directional drilling, 2002 m. long and 25 m. deep was a great challenge for Transierra - Owner - as well as Laney - Bolinter - Contractor - to accomplish a feat yet to be done in the entire world. The dedication of the people involved showed their degree of professionalism that these companies have obtained and the determination in doing the job overcoming unforeseen obstacles and still being able to finish on time, mitigating environmental impacts and leaving a first class crossing. This document presents a description of the technical, logistic and construction factors that were involved in the project and which allowed to perform 7 directional drillings, including Rio Grande River, which during the pull got the last 60 m. of pipe stuck, being freed only after using a pneumatic hammer. (author)

Human embryo-conditioned medium stimulates in vitro endometrial angiogenesis

NARCIS (Netherlands)

Kapiteijn, K.; Koolwijk, P.; Weiden, R.M.F. van der; Nieuw Amerongen, G. van; Plaisier, M.; Hinsbergh, V.W.M. van; Helmerhorst, F.M.

2006-01-01

Objective: Successful implantation and placentation depend on the interaction between the endometrium and the embryo. Angiogenesis is crucial at this time. In this article we investigate the direct influence of the human embryo on in vitro endometrial angiogenesis. Design: In vitro study. Setting:

Direct Utilization of Liquid Fuels in SOFC for Portable Applications: Challenges for the Selection of Alternative Anodes

Directory of Open Access Journals (Sweden)

Massimiliano Cimenti

2009-06-01

Full Text Available Solid oxide fuel cells (SOFC have the advantage of being able to operate with fuels other than hydrogen. In particular, liquid fuels are especially attractive for powering portable applications such as small power generators or auxiliary power units, in which case the direct utilization of the fuel would be convenient. Although liquid fuels are easier to handle and transport than hydrogen, their direct use in SOFC can lead to anode deactivation due to carbon formation, especially on traditional nickel/yttria stabilized zirconia (Ni/YSZ anodes. Significant advances have been made in anodic materials that are resistant to carbon formation but often these materials are less electrochemically active than Ni/YSZ. In this review the challenges of using liquid fuels directly in SOFC, in terms of gas-phase and catalytic reactions within the anode chamber, will be discussed and the alternative anode materials so far investigated will be compared.

Implementation challenges for designing integrated in vitro testing strategies (ITS) aiming at reducing and replacing animal experimentation.

Science.gov (United States)

De Wever, Bart; Fuchs, Horst W; Gaca, Marianna; Krul, Cyrille; Mikulowski, Stan; Poth, Albrecht; Roggen, Erwin L; Vilà, Maya R

2012-04-01

At the IVTIP (in vitro testing industrial platform) meeting of November 26th 2009 entitled 'Toxicology in the 21st century ('21C')--working our way towards a visionary reality' all delegates endorsed the emerging concept of the '21C' vision as the way forward to enable a thorough, reliable and systematic approach to future toxicity testing without the use of animals. One of the emerging concepts focused on integrating a defined number of tests modelling in vivo-relevant and well-characterised toxicity pathways representing mechanistic endpoints. At this meeting the importance of Integrated Testing Strategies (ITS) as tools towards reduction and eventually replacement of the animals currently used for hazard identification and risk assessment was recognised. A follow-up IVTIP Spring 2010 meeting entitled 'Integrated In Vitro Testing Strategies (ITS)--Implementation Challenges' was organised to address pending questions about ITS. This report is not a review of the ITS literature, but a summary of the discussions triggered by presented examples on how to develop and implement ITS. Contrasts between pharmaceutical and chemical industry, as well as a list of general but practical aspects to be considered while developing an ITS emerged from the discussions. In addition, current recommendations on the validation of ITS were discussed. In conclusion, the outcome of this workshop improved the understanding of the participants of some important factors that may impact the design of an ITS in function of its purpose (e.g., screening, or early decision making versus regulatory), the context in which they need to be applied (e.g., ICH guidelines, REACH) and the status and quality of the available tools. A set of recommendations of best practices was established and the importance of the applicability of the individual tests as well as the testing strategy itself was highlighted. Copyright © 2012 Elsevier Ltd. All rights reserved.

Evaluation of the in vitro stability of gadolinium (III) polyoxometalates

International Nuclear Information System (INIS)

Crooks, William J.; Choppin, Gregory R.; Rogers, Buck E.; Welch, Michael

1997-01-01

The gadolinium chelates of lacunary polyoxometalates were evaluated for in vitro stability against rat serum, diethylenetriaminepentaacetic acid (DTPA), endogenous metal cations, and DTPA-doped rat serum. The chelates dissociated rapidly in rat serum. Challenges by DTPA gave relatively slower dissociation rates, whereas challenges by endogenous metal cations (Fe(III), Zn(II), and Cu(II)) occurred at a rate comparable to the serum challenge, suggesting the instability in serum is due to a transmetalation mechanism. Challenges by DTPA-doped serum gave slower rates of dissociation than in native serum, verifying the transmetalation mechanism

Estimating Likelihood of Fetal In Vivo Interactions Using In Vitro HTS Data (Teratology meeting)

Science.gov (United States)

Tox21/ToxCast efforts provide in vitro concentration-response data for thousands of compounds. Predicting whether chemical-biological interactions observed in vitro will occur in vivo is challenging. We hypothesize that using a modified model from the FDA guidance for drug intera...

Material challenges for solar cells in the twenty-first century: directions in emerging technologies.

Science.gov (United States)

Almosni, Samy; Delamarre, Amaury; Jehl, Zacharie; Suchet, Daniel; Cojocaru, Ludmila; Giteau, Maxime; Behaghel, Benoit; Julian, Anatole; Ibrahim, Camille; Tatry, Léa; Wang, Haibin; Kubo, Takaya; Uchida, Satoshi; Segawa, Hiroshi; Miyashita, Naoya; Tamaki, Ryo; Shoji, Yasushi; Yoshida, Katsuhisa; Ahsan, Nazmul; Watanabe, Kentaro; Inoue, Tomoyuki; Sugiyama, Masakazu; Nakano, Yoshiaki; Hamamura, Tomofumi; Toupance, Thierry; Olivier, Céline; Chambon, Sylvain; Vignau, Laurence; Geffroy, Camille; Cloutet, Eric; Hadziioannou, Georges; Cavassilas, Nicolas; Rale, Pierre; Cattoni, Andrea; Collin, Stéphane; Gibelli, François; Paire, Myriam; Lombez, Laurent; Aureau, Damien; Bouttemy, Muriel; Etcheberry, Arnaud; Okada, Yoshitaka; Guillemoles, Jean-François

2018-01-01

Photovoltaic generation has stepped up within the last decade from outsider status to one of the important contributors of the ongoing energy transition, with about 1.7% of world electricity provided by solar cells. Progress in materials and production processes has played an important part in this development. Yet, there are many challenges before photovoltaics could provide clean, abundant, and cheap energy. Here, we review this research direction, with a focus on the results obtained within a Japan-French cooperation program, NextPV, working on promising solar cell technologies. The cooperation was focused on efficient photovoltaic devices, such as multijunction, ultrathin, intermediate band, and hot-carrier solar cells, and on printable solar cell materials such as colloidal quantum dots.

Biologically Based Methods for Pest Management in Agriculture under Changing Climates: Challenges and Future Directions.

Science.gov (United States)

Chidawanyika, Frank; Mudavanhu, Pride; Nyamukondiwa, Casper

2012-11-09

The current changes in global climatic regimes present a significant societal challenge, affecting in all likelihood insect physiology, biochemistry, biogeography and population dynamics. With the increasing resistance of many insect pest species to chemical insecticides and an increasing organic food market, pest control strategies are slowly shifting towards more sustainable, ecologically sound and economically viable options. Biologically based pest management strategies present such opportunities through predation or parasitism of pests and plant direct or indirect defense mechanisms that can all be important components of sustainable integrated pest management programs. Inevitably, the efficacy of biological control systems is highly dependent on natural enemy-prey interactions, which will likely be modified by changing climates. Therefore, knowledge of how insect pests and their natural enemies respond to climate variation is of fundamental importance in understanding biological insect pest management under global climate change. Here, we discuss biological control, its challenges under climate change scenarios and how increased global temperatures will require adaptive management strategies to cope with changing status of insects and their natural enemies.

Biologically Based Methods for Pest Management in Agriculture under Changing Climates: Challenges and Future Directions

Directory of Open Access Journals (Sweden)

Casper Nyamukondiwa

2012-11-01

Full Text Available The current changes in global climatic regimes present a significant societal challenge, affecting in all likelihood insect physiology, biochemistry, biogeography and population dynamics. With the increasing resistance of many insect pest species to chemical insecticides and an increasing organic food market, pest control strategies are slowly shifting towards more sustainable, ecologically sound and economically viable options. Biologically based pest management strategies present such opportunities through predation or parasitism of pests and plant direct or indirect defense mechanisms that can all be important components of sustainable integrated pest management programs. Inevitably, the efficacy of biological control systems is highly dependent on natural enemy-prey interactions, which will likely be modified by changing climates. Therefore, knowledge of how insect pests and their natural enemies respond to climate variation is of fundamental importance in understanding biological insect pest management under global climate change. Here, we discuss biological control, its challenges under climate change scenarios and how increased global temperatures will require adaptive management strategies to cope with changing status of insects and their natural enemies.

Dissecting direct and indirect readout of cAMP receptor protein DNA binding using an inosine and 2,6-diaminopurine in vitro selection system

DEFF Research Database (Denmark)

Lindemose, Søren; Nielsen, Peter E.; Møllegaard, Niels Erik

2008-01-01

The DNA interaction of the Escherichia coli cyclic AMP receptor protein (CRP) represents a typical example of a dual recognition mechanism exhibiting both direct and indirect readout. We have dissected the direct and indirect components of DNA recognition by CRP employing in vitro selection...... is functionally intact. The majority of the selected sites contain the natural consensus sequence TGTGAN(6)TCACA (i.e. TITIDN(6)TCDCD). Thus, direct readout of the consensus sequence is independent of minor groove conformation. Consequently, the indirect readout known to occur in the TG/CA base pair step (primary...... kink site) in the consensus sequence is not affected by I-D substitutions. In contrast, the flanking regions are selected as I/C rich sequences (mostly I-tracts) instead of A/T rich sequences which are known to strongly increase CRP binding, thereby demonstrating almost exclusive indirect readout...

In vitro transcription accurately predicts lac repressor phenotype in vivo in Escherichia coli

Directory of Open Access Journals (Sweden)

Matthew Almond Sochor

2014-07-01

Full Text Available A multitude of studies have looked at the in vivo and in vitro behavior of the lac repressor binding to DNA and effector molecules in order to study transcriptional repression, however these studies are not always reconcilable. Here we use in vitro transcription to directly mimic the in vivo system in order to build a self consistent set of experiments to directly compare in vivo and in vitro genetic repression. A thermodynamic model of the lac repressor binding to operator DNA and effector is used to link DNA occupancy to either normalized in vitro mRNA product or normalized in vivo fluorescence of a regulated gene, YFP. An accurate measurement of repressor, DNA and effector concentrations were made both in vivo and in vitro allowing for direct modeling of the entire thermodynamic equilibrium. In vivo repression profiles are accurately predicted from the given in vitro parameters when molecular crowding is considered. Interestingly, our measured repressor–operator DNA affinity differs significantly from previous in vitro measurements. The literature values are unable to replicate in vivo binding data. We therefore conclude that the repressor-DNA affinity is much weaker than previously thought. This finding would suggest that in vitro techniques that are specifically designed to mimic the in vivo process may be necessary to replicate the native system.

Developing technology-enhanced active learning for medical education: challenges, solutions, and future directions.

Science.gov (United States)

McCoy, Lise; Pettit, Robin K; Lewis, Joy H; Bennett, Thomas; Carrasco, Noel; Brysacz, Stanley; Makin, Inder Raj S; Hutman, Ryan; Schwartz, Frederic N

2015-04-01

Growing up in an era of video games and Web-based applications has primed current medical students to expect rapid, interactive feedback. To address this need, the A.T. Still University-School of Osteopathic Medicine in Arizona (Mesa) has developed and integrated a variety of approaches using technology-enhanced active learning for medical education (TEAL-MEd) into its curriculum. Over the course of 3 years (2010-2013), the authors facilitated more than 80 implementations of games and virtual patient simulations into the education of 550 osteopathic medical students. The authors report on 4 key aspects of the TEAL-MEd initiative, including purpose, portfolio of tools, progress to date regarding challenges and solutions, and future directions. Lessons learned may be of benefit to medical educators at academic and clinical training sites who wish to implement TEAL-MEd activities.

Bacterial adhesion on direct and indirect dental restorative composite resins: An in vitro study on a natural biofilm.

Science.gov (United States)

Derchi, Giacomo; Vano, Michele; Barone, Antonio; Covani, Ugo; Diaspro, Alberto; Salerno, Marco

2017-05-01

Both direct and indirect techniques are used for dental restorations. Which technique should be preferred or whether they are equivalent with respect to bacterial adhesion is unclear. The purpose of this in vitro study was to determine the affinity of bacterial biofilm to dental restorative composite resins placed directly and indirectly. Five direct composite resins for restorations (Venus Diamond, Adonis, Optifil, Enamel Plus HRi, Clearfil Majesty Esthetic) and 3 indirect composite resins (Gradia, Estenia, Signum) were selected. The materials were incubated in unstimulated whole saliva for 1 day. The biofilms grown were collected and their bacterial cells counted. In parallel, the composite resin surface morphology was analyzed with atomic force microscopy. Both bacterial cell count and surface topography parameters were subjected to statistical analysis (α=.05). Indirect composite resins showed significantly lower levels than direct composite resins for bacterial cell adhesion, (Pcomposite resins (P>.05). However, within the indirect composite resins a significantly lower level was found for Gradia than Estenia or Signum (Pcomposite resin roughness and bacterial adhesion when the second and particularly the third-order statistical moments of the composite resin height distributions were considered. Indirect dental restorative composite resins were found to be less prone to biofilm adhesion than direct composite resins. A correlation of bacterial adhesion to surface morphology exists that is described by kurtosis; thus, advanced data analysis is required to discover possible insights into the biologic effects of morphology. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Symbiotic in vitro seed propagation of Dendrobium: fungal and bacterial partners and their influence on plant growth and development.

Science.gov (United States)

Teixeira da Silva, Jaime A; Tsavkelova, Elena A; Zeng, Songjun; Ng, Tzi Bun; Parthibhan, S; Dobránszki, Judit; Cardoso, Jean Carlos; Rao, M V

2015-07-01

The genus Dendrobium is one of the largest genera of the Orchidaceae Juss. family, although some of its members are the most threatened today. The reason why many species face a vulnerable or endangered status is primarily because of anthropogenic interference in natural habitats and commercial overexploitation. The development and application of modern techniques and strategies directed towards in vitro propagation of orchids not only increases their number but also provides a viable means to conserve plants in an artificial environment, both in vitro and ex vitro, thus providing material for reintroduction. Dendrobium seed germination and propagation are challenging processes in vivo and in vitro, especially when the extreme specialization of these plants is considered: (1) their biotic relationships with pollinators and mycorrhizae; (2) adaptation to epiphytic or lithophytic life-styles; (3) fine-scale requirements for an optimal combination of nutrients, light, temperature, and pH. This review also aims to summarize the available data on symbiotic in vitro Dendrobium seed germination. The influence of abiotic factors as well as composition and amounts of different exogenous nutrient substances is examined. With a view to better understanding how to optimize and control in vitro symbiotic associations, a part of the review describes the strong biotic relations of Dendrobium with different associative microorganisms that form microbial communities with adult plants, and also influence symbiotic seed germination. The beneficial role of plant growth-promoting bacteria is also discussed.

Metagenomic Sequencing of an In Vitro-Simulated Microbial Community

Energy Technology Data Exchange (ETDEWEB)

Morgan, Jenna L.; Darling, Aaron E.; Eisen, Jonathan A.

2009-12-01

Background: Microbial life dominates the earth, but many species are difficult or even impossible to study under laboratory conditions. Sequencing DNA directly from the environment, a technique commonly referred to as metagenomics, is an important tool for cataloging microbial life. This culture-independent approach involves collecting samples that include microbes in them, extracting DNA from the samples, and sequencing the DNA. A sample may contain many different microorganisms, macroorganisms, and even free-floating environmental DNA. A fundamental challenge in metagenomics has been estimating the abundance of organisms in a sample based on the frequency with which the organism's DNA was observed in reads generated via DNA sequencing. Methodology/Principal Findings: We created mixtures of ten microbial species for which genome sequences are known. Each mixture contained an equal number of cells of each species. We then extracted DNA from the mixtures, sequenced the DNA, and measured the frequency with which genomic regions from each organism was observed in the sequenced DNA. We found that the observed frequency of reads mapping to each organism did not reflect the equal numbers of cells that were known to be included in each mixture. The relative organism abundances varied significantly depending on the DNA extraction and sequencing protocol utilized. Conclusions/Significance: We describe a new data resource for measuring the accuracy of metagenomic binning methods, created by in vitro-simulation of a metagenomic community. Our in vitro simulation can be used to complement previous in silico benchmark studies. In constructing a synthetic community and sequencing its metagenome, we encountered several sources of observation bias that likely affect most metagenomic experiments to date and present challenges for comparative metagenomic studies. DNA preparation methods have a particularly profound effect in our study, implying that samples prepared with

Fetal Cardiac Doppler Signal Processing Techniques: Challenges and Future Research Directions

Directory of Open Access Journals (Sweden)

Saeed Abdulrahman Alnuaimi

2017-12-01

Full Text Available The fetal Doppler Ultrasound (DUS is commonly used for monitoring fetal heart rate and can also be used for identifying the event timings of fetal cardiac valve motions. In early-stage fetuses, the detected Doppler signal suffers from noise and signal loss due to the fetal movements and changing fetal location during the measurement procedure. The fetal cardiac intervals, which can be estimated by measuring the fetal cardiac event timings, are the most important markers of fetal development and well-being. To advance DUS-based fetal monitoring methods, several powerful and well-advanced signal processing and machine learning methods have recently been developed. This review provides an overview of the existing techniques used in fetal cardiac activity monitoring and a comprehensive survey on fetal cardiac Doppler signal processing frameworks. The review is structured with a focus on their shortcomings and advantages, which helps in understanding fetal Doppler cardiogram signal processing methods and the related Doppler signal analysis procedures by providing valuable clinical information. Finally, a set of recommendations are suggested for future research directions and the use of fetal cardiac Doppler signal analysis, processing, and modeling to address the underlying challenges.

Material challenges for solar cells in the twenty-first century: directions in emerging technologies

Science.gov (United States)

Delamarre, Amaury; Jehl, Zacharie; Suchet, Daniel; Cojocaru, Ludmila; Giteau, Maxime; Behaghel, Benoit; Julian, Anatole; Ibrahim, Camille; Tatry, Léa; Wang, Haibin; Kubo, Takaya; Uchida, Satoshi; Segawa, Hiroshi; Miyashita, Naoya; Tamaki, Ryo; Shoji, Yasushi; Yoshida, Katsuhisa; Ahsan, Nazmul; Watanabe, Kentaro; Inoue, Tomoyuki; Sugiyama, Masakazu; Nakano, Yoshiaki; Hamamura, Tomofumi; Toupance, Thierry; Olivier, Céline; Chambon, Sylvain; Vignau, Laurence; Geffroy, Camille; Cloutet, Eric; Hadziioannou, Georges; Cavassilas, Nicolas; Rale, Pierre; Cattoni, Andrea; Collin, Stéphane; Gibelli, François; Paire, Myriam; Lombez, Laurent; Aureau, Damien; Bouttemy, Muriel; Etcheberry, Arnaud; Okada, Yoshitaka

2018-01-01

Abstract Photovoltaic generation has stepped up within the last decade from outsider status to one of the important contributors of the ongoing energy transition, with about 1.7% of world electricity provided by solar cells. Progress in materials and production processes has played an important part in this development. Yet, there are many challenges before photovoltaics could provide clean, abundant, and cheap energy. Here, we review this research direction, with a focus on the results obtained within a Japan–French cooperation program, NextPV, working on promising solar cell technologies. The cooperation was focused on efficient photovoltaic devices, such as multijunction, ultrathin, intermediate band, and hot-carrier solar cells, and on printable solar cell materials such as colloidal quantum dots. PMID:29707072

Facial Animations: Future Research Directions & Challenges

Science.gov (United States)

Alkawaz, Mohammed Hazim; Mohamad, Dzulkifli; Rehman, Amjad; Basori, Ahmad Hoirul

2014-06-01

Nowadays, computer facial animation is used in a significant multitude fields that brought human and social to study the computer games, films and interactive multimedia reality growth. Authoring the computer facial animation, complex and subtle expressions are challenging and fraught with problems. As a result, the current most authored using universal computer animation techniques often limit the production quality and quantity of facial animation. With the supplement of computer power, facial appreciative, software sophistication and new face-centric methods emerging are immature in nature. Therefore, this paper concentrates to define and managerially categorize current and emerged surveyed facial animation experts to define the recent state of the field, observed bottlenecks and developing techniques. This paper further presents a real-time simulation model of human worry and howling with detail discussion about their astonish, sorrow, annoyance and panic perception.

Nanobiotechnology approach using plant rooting hormone synthesized silver nanoparticle as “nanobullets” for the dynamic applications in horticulture – An in vitro and ex vitro study

Directory of Open Access Journals (Sweden)

Raja Muthuramalingam Thangavelu

2018-01-01

Full Text Available Horticulture is the branch of agriculture that deals with science and technology and business of plant cultivation and it is considered to be the foremost part of the world economy. Even though, one of the major challenges which has seriously influenced the economic loss of horticulture is rooting of cuttings and root growth inhibiting plant pathogens. To address this issue through nanobiotechnology, we ingeniously build a concept of silver nanoparticles (AgNPs as “nano-bullets” can act for a dual mode like root enhancer and pathogen destroyer on the target site. After that, we succeeded in AgNPs synthesis, using two auxin rooting hormones of Indole-3-acetic acid and Indole-3-butyric acid as a reducing cum stabilizing agent. Further, its efficacy of root promoting and pathogen inhibitory action was sufficiently validated through in vitro and ex vitro studies with model plants and plant pathogens. As a result, the action duality of hormone-stabilized AgNPs was manifested to threefold enhanced root growth compared to controls and it increased the rooting capabilities against root growth inhibiting phytopathogens. This feature was also proved by the direct antifungal assay. Moreover, hormone-AgNPs left no toxicity to treated plants which was revealed by RAPD molecular markers. Therefore, with a detailed study and analysis with instruments such as Spectroscopy, TEM, Zetasizer, FTIR, Cyclic Voltammetry, Fluorescence microscopy (nanoparticles uptake, SEM coupled with EDS (bioaccumulation, TGA (grafting density and PCR (RAPD analysis, this study can unravel the relevance, scope and current challenges at horticulture plants root development and plant disease management for the sustainable agricultural crop production.

Targeted in vitro and in vivo gene transfer into T Lymphocytes: potential of direct inhibition of allo-immune activation

Directory of Open Access Journals (Sweden)

Mehra Mandeep R

2006-11-01

Full Text Available Abstract Background Successful inhibition of alloimmune activation in organ transplantation remains one of the key events in achieving a long-term graft survival. Since T lymphocytes are largely responsible for alloimmune activation, targeted gene transfer of gene of cyclin kinase inhibitor p21 into T cells might inhibit their aberrant proliferation. A number of strategies using either adenoviral or lentiviral vectors linked to mono or bispecific antibodies directed against T cell surface markers/cytokines did not yield the desired results. Therefore, this study was designed to test if a CD3promoter-p21 chimeric construct would in vitro and in vivo transfer p21 gene to T lymphocytes and result in inhibition of proliferation. CD3 promoter-p21 chimeric constructs were prepared with p21 in the sense and antisense orientation. For in vitro studies EL4-IL-2 thyoma cells were used and for in vivo studies CD3p21 sense and antisense plasmid DNA was injected intramuscularly in mice. Lymphocyte proliferation was quantified by 3H-thymidine uptake assay; IL-2 mRNA expression was studied by RT-PCR and using Real Time PCR assay, we monitored the CD3, p21, TNF-α and IFN-γ mRNA expression. Results Transfection of CD3p21 sense and antisense in mouse thyoma cell line (EL4-IL-2 resulted in modulation of mitogen-induced proliferation. The intramuscular injection of CD3p21 sense and antisense plasmid DNA into mice also modulated lymphocyte proliferation and mRNA expression of pro-inflammatory cytokines. Conclusion These results demonstrate a novel strategy of in vitro and in vivo transfer of p21 gene to T cells using CD3-promoter to achieve targeted inhibition of lymphocyte proliferation and immune activation.

In vitro models to evaluate the permeability of poorly soluble drug entities: Challenges and perspectives

DEFF Research Database (Denmark)

Buckley, S. T.; Fischer, S. M.; Fricker, G.

2012-01-01

The application of in vitro models in drug permeability studies represents a useful screening tool for assessing the biopharmaceutical appropriateness of new chemical entities (NCEs). Of note, there remains an ever-increasing number of NCEs which exhibit poor aqueous solubility. However, in their......The application of in vitro models in drug permeability studies represents a useful screening tool for assessing the biopharmaceutical appropriateness of new chemical entities (NCEs). Of note, there remains an ever-increasing number of NCEs which exhibit poor aqueous solubility. However...

Coupling of In Vitro Bioassays with Planar Chromatography in Effect-Directed Analysis.

Science.gov (United States)

Weiss, Stefan C; Egetenmeyer, Nicole; Schulz, Wolfgang

Modern analytical test methods increasingly detect anthropogenic organic substances and their transformation products in water samples and in the environment. The presence of these compounds might pose a risk to the aquatic environment. To determine a possible (eco)toxicological risk, aquatic samples are tested using various bioassays, including sub-organismic assays such as the luminescent bacteria inhibition test, the acetylcholinesterase inhibition test, and the umu-test. The effect-directed analysis (EDA) combines physicochemical separation methods with biological (in vitro) tests. High-performance thin-layer chromatography (HPTLC) has proved to be particularly well suited for the separation of organic compounds and the subsequent analysis of effects by the application of the biotests directly on the surface of the HPTLC plate. The advantage of using HPTLC in comparison to high-performance liquid chromatography (HPLC) for EDA is that the solvent which is used as a mobile phase during chromatography is completely evaporated after the separation and therefore can no longer influence the applied bioassays.A prioritization during the complex identification process can be achieved when observed effects are associated with the separated zones in HPTLC. This increases the probability of identifying the substance responsible for an adverse effect from the multitude of organic trace substances in environmental samples. Furthermore, by comparing the pattern of biological effects of a separated sample, it is possible to track and assess changes in biological activity over time, over space, or in the course of a process, even without identifying the substance. HPTLC has already been coupled with various bioassays.Because HPTLC is a very flexible system, various detection techniques can be used and combined. In addition to the UV/Vis absorption and fluorescence measurements, TLC can also be coupled with a mass spectrometer (MS) for compound identification. In addition

biomedical challenges of human senescence: a review

African Journals Online (AJOL)

2002-12-12

Dec 12, 2002 ... generation per mitochondrion. Understanding the role of cellular ageing in vivo is another major challenge for integrative study. The in vitro model of cell replicative senescence pioneered by. Sith and Pereira has elucidated the mechanisms limiting cell proliferation(30). Until recently the link between in.

Opportunities and challenges in green house gases reduction using high pressure direct injection of natural gas

International Nuclear Information System (INIS)

Ouellette, P.

2001-01-01

In an effort to reduce Greenhouse Gases, Westport Innovations is developing a high pressure direct injection (HPDI) technology for gaseous fuels. This technology adapts the diesel cycle for gaseous fuels, since the diesel cycle provides high efficiency, high low-speed torque, fast transient capabilities and reliability. Because of their high efficiency, diesels are very favorable from a Greenhouse Gas (GHG) point of view, however they remain challenged by high nitrogen oxides (NOx) and particulate matter (PM) emissions. When directly injecting natural gas, NOx and PM emissions can be reduced by approximately 50% while maintaining the performance of the diesel engine. This allows the use of abundant and historically cheaper natural gas. Because of its lower carbon content per unit energy, natural gas also offers further GHG reduction over the diesel if the efficiency is preserved and if methane emissions are low. This paper discusses development efforts at Westport for several applications including on-highway trucks, light-duty delivery trucks and power generation

Routing Protocols for Underwater Wireless Sensor Networks: Taxonomy, Research Challenges, Routing Strategies and Future Directions.

Science.gov (United States)

Khan, Anwar; Ali, Ihsan; Ghani, Abdullah; Khan, Nawsher; Alsaqer, Mohammed; Rahman, Atiq Ur; Mahmood, Hasan

2018-05-18

merit(s). Finally, open challenges and research directions are presented for future investigation.

An in vitro tag-and-modify protein sample generation method for single-molecule fluorescence resonance energy transfer.

Science.gov (United States)

Hamadani, Kambiz M; Howe, Jesse; Jensen, Madeleine K; Wu, Peng; Cate, Jamie H D; Marqusee, Susan

2017-09-22

Biomolecular systems exhibit many dynamic and biologically relevant properties, such as conformational fluctuations, multistep catalysis, transient interactions, folding, and allosteric structural transitions. These properties are challenging to detect and engineer using standard ensemble-based techniques. To address this drawback, single-molecule methods offer a way to access conformational distributions, transient states, and asynchronous dynamics inaccessible to these standard techniques. Fluorescence-based single-molecule approaches are parallelizable and compatible with multiplexed detection; to date, however, they have remained limited to serial screens of small protein libraries. This stems from the current absence of methods for generating either individual dual-labeled protein samples at high throughputs or protein libraries compatible with multiplexed screening platforms. Here, we demonstrate that by combining purified and reconstituted in vitro translation, quantitative unnatural amino acid incorporation via AUG codon reassignment, and copper-catalyzed azide-alkyne cycloaddition, we can overcome these challenges for target proteins that are, or can be, methionine-depleted. We present an in vitro parallelizable approach that does not require laborious target-specific purification to generate dual-labeled proteins and ribosome-nascent chain libraries suitable for single-molecule FRET-based conformational phenotyping. We demonstrate the power of this approach by tracking the effects of mutations, C-terminal extensions, and ribosomal tethering on the structure and stability of three protein model systems: barnase, spectrin, and T4 lysozyme. Importantly, dual-labeled ribosome-nascent chain libraries enable single-molecule co-localization of genotypes with phenotypes, are well suited for multiplexed single-molecule screening of protein libraries, and should enable the in vitro directed evolution of proteins with designer single-molecule conformational

Advance Directives in Hospice Healthcare Providers: A Clinical Challenge.

Science.gov (United States)

Luck, George R; Eggenberger, Terry; Newman, David; Cortizo, Jacqueline; Blankenship, Derek C; Hennekens, Charles H

2017-11-01

On a daily basis, healthcare providers, especially those dealing with terminally ill patients, such as hospice workers, witness how advance directives help ensure the wishes of patients. They also witness the deleterious consequences when patients fail to document the care they desire at their end of life. To the best of our knowledge there are no data concerning the prevalence of advance directives among hospice healthcare providers. We therefore explored the prevalence and factors influencing completion rates in a survey of hospice healthcare providers. Surveys that included 32 items to explore completion rates, as well as barriers, knowledge, and demographics, were e-mailed to 2097 healthcare providers, including employees and volunteers, at a nonprofit hospice. Of 890 respondents, 44% reported having completed an advance directive. Ethnicity, age, relationship status, and perceived knowledge were all significant factors influencing the completion rates, whereas years of experience or working directly with patients had no effect. Procrastination, fear of the subject, and costs were common reasons reported as barriers. Upon completion of the survey, 43% said they will now complete an advance directive, and 45% will talk to patients and families about their wishes. The majority of hospice healthcare providers have not completed an advance directive. These results are very similar to those for other healthcare providers treating patients with terminal diseases, specifically oncologists. Because, at completion, 43% said that they would now complete an advance directive, such a survey of healthcare providers may help increase completion rates. Copyright © 2017 Elsevier Inc. All rights reserved.

Key Challenges and Opportunities Associated with the Use of In Vitro Models to Detect Human DILI: Integrated Risk Assessment and Mitigation Plans

Directory of Open Access Journals (Sweden)

Franck A. Atienzar

2016-01-01

Full Text Available Drug-induced liver injury (DILI is a major cause of late-stage clinical drug attrition, market withdrawal, black-box warnings, and acute liver failure. Consequently, it has been an area of focus for toxicologists and clinicians for several decades. In spite of considerable efforts, limited improvements in DILI prediction have been made and efforts to improve existing preclinical models or develop new test systems remain a high priority. While prediction of intrinsic DILI has improved, identifying compounds with a risk for idiosyncratic DILI (iDILI remains extremely challenging because of the lack of a clear mechanistic understanding and the multifactorial pathogenesis of idiosyncratic drug reactions. Well-defined clinical diagnostic criteria and risk factors are also missing. This paper summarizes key data interpretation challenges, practical considerations, model limitations, and the need for an integrated risk assessment. As demonstrated through selected initiatives to address other types of toxicities, opportunities exist however for improvement, especially through better concerted efforts at harmonization of current, emerging and novel in vitro systems or through the establishment of strategies for implementation of preclinical DILI models across the pharmaceutical industry. Perspectives on the incorporation of newer technologies and the value of precompetitive consortia to identify useful practices are also discussed.

In vitro propagation of persimmon (Diospyros kaki Thunb.).

Science.gov (United States)

Giordani, Edgardo; Naval, Mar; Benelli, Carla

2013-01-01

Persimmon (Diospyros kaki Thunb.) is a temperate fruit tree species diffused in all continents. The traditional propagation method adopted by the nursery industry is based on budding/grafting scion cultivars on seedlings from D. kaki, Diospyros lotus, and Diospyros virginiana, the most important species used as rootstock, reproduced by seeds since they are not easy to root. Furthermore, most of nonastringent cultivars of persimmon are not compatible with D. lotus, a rootstock largely utilized because of its hardiness and frost resistance. The main in vitro tissue culture techniques, developed for persimmon, deal with direct regeneration (from dormant buds and root tips), and indirect regeneration through callus from dormant buds, apexes, and leaves. The bottlenecks of micropropagation are (1) the recalcitrance of many cultivars to in vitro establishment, (2) the low multiplication ratio of D. kaki compared to other fruit tree species, (3) the very low rooting ability of ex novo microcuttings both from direct and indirect regeneration, (4) the high sensitivity to transplant from in vitro to in vivo conditions. The development of reliable in vitro regeneration procedures is likely to play a key role for production of both clonal rootstocks and self-rooted cultivars. The general protocol for micropropagation of persimmon reported here is based on the establishment of winter dormant buds in vitro, shoot development, multiplication and elongation, and shoot rooting, using cytokinins (BA or zeatin) in a MS media along with an auxinic pretreatment for rooting induction.

In vitro growth of Plasmodium falciparum in neonatal blood.

Science.gov (United States)

Sauerzopf, Ulrich; Honkpehedji, Yabo J; Adgenika, Ayôla A; Feugap, Elianne N; Ngoma, Ghyslain Mombo; Mackanga, Jean-Rodolphe; Lötsch, Felix; Loembe, Marguerite M; Kremsner, Peter G; Mordmüller, Benjamin; Ramharter, Michael

2014-11-18

Children below the age of six months suffer less often from malaria than older children in sub-Saharan Africa. This observation is commonly attributed to the persistence of foetal haemoglobin (HbF), which is considered not to permit growth of Plasmodium falciparum and therefore providing protection against malaria. Since this concept has recently been challenged, this study evaluated the effect of HbF erythrocytes and maternal plasma on in vitro parasite growth of P. falciparum in Central African Gabon. Umbilical cord blood and peripheral maternal blood were collected at delivery at the Albert Schweitzer Hospital in Gabon. Respective erythrocyte suspension and plasma were used in parallel for in vitro culture. In vitro growth rates were compared between cultures supplemented with either maternal or cord erythrocytes. Plasma of maternal blood and cord blood was evaluated. Parasite growth rates were assessed by the standard HRP2-assay evaluating the increase of HRP2 concentration in Plasmodium culture. Culture of P. falciparum using foetal erythrocytes led to comparable growth rates (mean growth rate = 4.2, 95% CI: 3.5 - 5.0) as cultures with maternal red blood cells (mean growth rate =4.2, 95% CI: 3.4 - 5.0) and those from non-malaria exposed individuals (mean growth rate = 4.6, 95% CI: 3.8 - 5.5). Standard in vitro culture of P. falciparum supplemented with either maternal or foetal plasma showed both significantly lower growth rates than a positive control using non-malaria exposed donor plasma. These data challenge the concept of HbF serving as intrinsic inhibitor of P. falciparum growth in the first months of life. Erythrocytes containing HbF are equally permissive to P. falciparum growth in vitro. However, addition of maternal and cord plasma led to reduced in vitro growth which may translate to protection against clinical disease or show synergistic effects with HbF in vivo. Further studies are needed to elucidate the pathophysiology of innate and acquired

In Vitro Exposure Systems and Dosimetry Assessment Tools ...

Science.gov (United States)

In 2009, the passing of The Family Smoking Prevention and Tobacco Control Act facilitated the establishment of the FDA Center for Tobacco Products (CTP) and gave it regulatory authority over the marketing, manufacture and distribution of tobacco products, including those termed “modified risk”. On 4-6 April 2016, the Institute for In Vitro Sciences, Inc. (IIVS) convened a workshop conference titled “In Vitro Exposure Systems and Dosimetry Assessment Tools for Inhaled Tobacco Products” to bring together stakeholders representing regulatory agencies, academia, and industry to address the research priorities articulated by the FDA CTP. Specific topics were covered to assess the status of current in vitro smoke and aerosol/vapor exposure systems, as well as the various approaches and challenges to quantifying the complex exposures, in in vitro pulmonary models developed for evaluating adverse pulmonary events resulting from tobacco product exposures. The four core topics covered were, 1) Tobacco Smoke And E-Cigarette Aerosols, 2) Air-Liquid Interface-In Vitro Exposure Systems, 3) Dosimetry Approaches For Particles And Vapors; In Vitro Dosimetry Determinations and 4) Exposure Microenvironment/Physiology Of Cells. The two and a half day workshop included presentations from 20 expert speakers, poster sessions, networking discussions, and breakout sessions which identified key findings and provided recommendations to advance these technologies. Here, we will re

Critical analysis of 3-D organoid in vitro cell culture models for high-throughput drug candidate toxicity assessments.

Science.gov (United States)

Astashkina, Anna; Grainger, David W

2014-04-01

Drug failure due to toxicity indicators remains among the primary reasons for staggering drug attrition rates during clinical studies and post-marketing surveillance. Broader validation and use of next-generation 3-D improved cell culture models are expected to improve predictive power and effectiveness of drug toxicological predictions. However, after decades of promising research significant gaps remain in our collective ability to extract quality human toxicity information from in vitro data using 3-D cell and tissue models. Issues, challenges and future directions for the field to improve drug assay predictive power and reliability of 3-D models are reviewed. Copyright © 2014 Elsevier B.V. All rights reserved.

In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish challenged with GFP tagged Vibrio parahaemolyticus Dahv2.

Science.gov (United States)

Girija, Vairavan; Malaikozhundan, Balasubramanian; Vaseeharan, Baskaralingam; Vijayakumar, Sekar; Gobi, Narayanan; Del Valle Herrera, Marian; Chen, Jiann-Chu; Santhanam, Perumal

2018-01-01

In vitro antagonistic activity and the protective effect of probiotic Bacillus licheniformis Dahb1 in zebrafish (Danio rerio) challenged with GFP tagged Vibrio parahaemolyticus Dahv2 was studied. The cell free extract of probiotic B. licheniformis Dahb1 at 100 μg mL -1 showed growth inhibition of V. parahaemolyticus Dahv2 in vitro. B. licheniformis Dahb1 also inhibited the biofilm growth of GFP tagged V. parahaemolyticus Dahv2 at 100 μg mL -1 in vitro. The growth and survival of zebrafish was tested using probiotic B. licheniformis Dahb1. Weight (1.28 g) of zebrafish that received the cell free extract was much higher than in control (1.04 g). The mortality of zebrafish infected with GFP tagged V. parahaemolyticus Dahv2 at 10 7 Cfu mL -1 (Group IV) was 100%, whereas a complete survival of zebrafish that received the cell free extract of B. licheniformis Dahb1 at 10 7 Cfu mL -1 (Group VII) was observed after 30 days. The number of GFP tagged V. parahaemolyticus Dahv2 colonies in the intestine and gills significantly reduced after treatment with the cell free extract of B. licheniformis Dahb1. Furthermore, a significant decrease in the fluorescent colonies of GFP tagged V. parahaemolyticus Dahv2 was observed after treatment with the cell free extract of B. licheniformis Dahb1 under confocal laser scanning microscopy (CLSM). In conclusion, the cell free extract of B. licheniformis Dahb1 could prevent Vibrio infection by enhancing the growth and survival of zebrafish. Copyright © 2017 Elsevier Ltd. All rights reserved.

Oral food challenges in children

Directory of Open Access Journals (Sweden)

Hye Yung Yum

2011-01-01

Full Text Available Many patients assume that allergic reactions against foods are responsible for triggering or worsening their allergic symptoms. Therefore, it is important to identify patients who would benefit from an elimination diet, while avoiding unnecessary dietary restrictions. The diagnosis of food allergy depends on the thorough review of the patients's medical history, results of supplemented trials of dietary elimination, and in vivo and in vitro tests for measuring specific IgE levels. However, in some cases the reliability of such procedures is suboptimal. Oral food challenges are procedures employed for making an accurate diagnosis of immediate and occasionally delayed adverse reactions to foods. The timing and type of the challenge, preparation of patients, foods to be tested, and dosing schedule should be determined on the basis of the patient's history, age, and experience. Although double-blind, placebo-controlled food challenges(DBPCFC are used to establish definitively if a food is the cause of adverse reactions, they are time-consuming, expensive and troublesome for physician and patients. In practice, An open challenge controlled by trained personnel is sufficient especially in infants and young children. The interpretation of the results and follow-up after a challenge are also important. Since theses challenges are relatively safe and informative, controlled oral food challenges could become the measure of choice in children.

Blockchain-oriented Software Engineering: Challenges and New Directions

OpenAIRE

Porru, Simone; Pinna, Andrea; Marchesi, Michele; Tonelli, Roberto

2017-01-01

The Blockchain technology is reshaping finance, economy, money to the extent that its disruptive power is compared to that of the Internet and the Web in their early days. As a result, all the software development revolving around the Blockchain technology is growing at a staggering rate. In this paper, we acknowledge the need for software engineers to devise specialized tools and techniques for blockchain-oriented software development. From current challenges concerning the definition of new...

Challenges to develop nitrogen-fixing cereals by direct nif-gene transfer.

Science.gov (United States)

Curatti, Leonardo; Rubio, Luis M

2014-08-01

Some regions of the developing world suffer low cereal production yields due to low fertilizer inputs, among other factors. Biological N2 fixation, catalyzed by the prokaryotic enzyme nitrogenase, is an alternative to the use of synthetic N fertilizers. The molybdenum nitrogenase is an O2-labile metalloenzyme composed of the NifDK and NifH proteins, which biosyntheses require a number of nif gene products. A challenging strategy to increase cereal crop productivity in a scenario of low N fertilization is the direct transfer of nif genes into cereals. The sensitivity of nitrogenase to O2 and the apparent complexity of nitrogenase biosynthesis are the main barriers identified so far. Expression of active NifH requires the products of nifM, nifH, and possibly nifU and nifS, whereas active NifDK requires the products of nifH, nifD, nifK, nifB, nifE, nifN, and possibly nifU, nifS, nifQ, nifV, nafY, nifW and nifZ. Plastids and mitochondria are potential subcellular locations for nitrogenase. Both could provide the ATP and electrons required for nitrogenase to function but they differ in their internal O2 levels and their ability to incorporate ammonium into amino acids. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Methodological challenges surrounding direct-to-consumer advertising research--the measurement conundrum.

Science.gov (United States)

Hansen, Richard A; Droege, Marcus

2005-06-01

Numerous studies have focused on the impact of direct-to-consumer (DTC) prescription drug advertising on consumer behavior and health outcomes. These studies have used various approaches to assess exposure to prescription drug advertising and to measure the subsequent effects of such advertisements. The objectives of this article are to (1) discuss measurement challenges involved in DTC advertising research, (2) summarize measurement approaches commonly identified in the literature, and (3) discuss contamination, time to action, and endogeneity as specific problems in measurement design and application. We conducted a review of the professional literature to identify illustrative approaches to advertising measurement. Specifically, our review of the literature focused on measurement of DTC advertising exposure and effect. We used the hierarchy-of-effects model to guide our discussion of processing and communication effects. Other effects were characterized as target audience action, sales, market share, and profit. Overall, existing studies have used a variety of approaches to measure advertising exposure and effect, yet the ability of measures to produce a valid and reliable understanding of the effects of DTC advertising can be improved. Our review provides a framework for conceptualizing DTC measurement, and can be used to identify gaps in the literature not sufficiently addressed by existing measures. Researchers should continue to explore correlations between exposure and effect of DTC advertising, but are obliged to improve and validate measurement in this area.

Learning Analytics: Challenges and Future Research Directions

Directory of Open Access Journals (Sweden)

Vlatko Lukarov

2014-12-01

Full Text Available In recent years, learning analytics (LA has attracted a great deal of attention in technology-enhanced learning (TEL research as practitioners, institutions, and researchers are increasingly seeing the potential that LA has to shape the future TEL landscape. Generally, LA deals with the development of methods that harness educational data sets to support the learning process. This paper provides a foundation for future research in LA. It provides a systematic overview on this emerging field and its key concepts through a reference model for LA based on four dimensions, namely data, environments, context (what?, stakeholders (who?, objectives (why?, and methods (how?. It further identifies various challenges and research opportunities in the area of LA in relation to each dimension.

In vitro evaluation of carbon-nanotube-reinforced bioprintable vascular conduits

International Nuclear Information System (INIS)

Dolati, Farzaneh; Yu, Yin; Zhang, Yahui; Ozbolat, Ibrahim T; Jesus, Aribet M De; Sander, Edward A

2014-01-01

Vascularization of thick engineered tissue and organ constructs like the heart, liver, pancreas or kidney remains a major challenge in tissue engineering. Vascularization is needed to supply oxygen and nutrients and remove waste in living tissues and organs through a network that should possess high perfusion ability and significant mechanical strength and elasticity. In this paper, we introduce a fabrication process to print vascular conduits directly, where conduits were reinforced with carbon nanotubes (CNTs) to enhance their mechanical properties and bioprintability. In vitro evaluation of printed conduits encapsulated in human coronary artery smooth muscle cells was performed to characterize the effects of CNT reinforcement on the mechanical, perfusion and biological performance of the conduits. Perfusion and permeability, cell viability, extracellular matrix formation and tissue histology were assessed and discussed, and it was concluded that CNT-reinforced vascular conduits provided a foundation for mechanically appealing constructs where CNTs could be replaced with natural protein nanofibers for further integration of these conduits in large-scale tissue fabrication. (paper)

Propagação in vitro de Sacha inchi In vitro propagation of Sacha inchi

Directory of Open Access Journals (Sweden)

Stevan Ricardo Bordignon

2012-07-01

Full Text Available O objetivo do presente trabalho foi avaliar in vitro a relação auxina:citocinina a fim de obter propágulos nos segmentos distintos do epicótilo e hipocótilo de sementes germinadas in vitro de Sacha inchi (Plukenetia volubilis Linneo. Os segmentos apical (A, mediano (B e o basal (C foram introduzidos em meio de cultivo MS, semi sólido (2,0g L-1 Phytagel, suplementado com vitaminas de MS, sacarose (30,0g L-1 e submetidos a três doses da auxina ácido indolbutírico - IBA (0; 0,1; 0,5mg L-1, associadas a quatro doses da citocinina benzilaminopurina - BAP (0; 0,1; 0,5; 1,0mg L-1, totalizando 36 tratamentos. Após nove semanas de cultivo in vitro, o segmento apical (A apresentou formação de brotações por organogênese direta nas concentrações de 0,5 e 1,0 de BAP, associado a 0,0 e 0,1 de IBA. O emprego do cultivo in vitro é viável na produção de mudas, utilizando como explante a região apical de sementes germinadas in vitro.The aim of this study was to perform an in vitro evaluation of the auxin:cytokinine ratio in different segments of the epicotyl and hypocotyl of Sacha inchi (Plukenetia volubilis Linneo seeds germinated in vitro. The segments apical (A, median (B and basal (C were introduced into semi-solid MS culture medium (2.0g L-1 Phytagel, supplemented with MS vitamins, sucrose (30.0g L-1 and submitted to three doses of auxin indolebutyric acid - IBA (0; 0.1; 0.5mg L-1, associated with four doses of the cytokinine benzylaminopurine - BAP (0; 0.1; 0.5; 1.0mg L-1, totaling 36 treatments. After nine weeks of in vitro cultivation, the apical segment (A presented shoot formation by direct organogenesis at the concentrations of 0.5 and 1.0 of BAP associated with 0.0 and 0.1 of IBA. It is feasible to use in vitro cultivation with the apical region of seeds germinated in vitro used as explants.

Parallelogram approach using rat-human In vitro and rat in vivo toxicogenomics predicts acetaminophen-induced hepatotoxicity in humans

NARCIS (Netherlands)

Kienhuis, A.S.; Poll, M.C.G. van de; Wortelboer, H.; Herwijnen, M. van; Gottschalk, R.; Jong, C.H.C. de; Boorsma, A.; Paules, R.S.; Kleinjans, J.C.S.; Stierum, R.H.; Delft, J.H.M. van

2009-01-01

The frequent use of rodent hepatic in vitro systems in pharmacological and toxicological investigations challenges extrapolation of in vitro results to the situation in vivo and interspecies extrapolation from rodents to humans. The toxicogenomics approach may aid in evaluating relevance of these

Direct contact between dendritic cells and bronchial epithelial cells inhibits T cell recall responses towards mite and pollen allergen extracts in vitro

DEFF Research Database (Denmark)

Papazian, Dick; Wagtmann, Valery R; Hansen, Soren

2015-01-01

(DCs), we have investigated recall T cell responses in allergic patients sensitized to house dust mite, grass, and birch pollen. Conclusions: Using allergen extract-loaded DCs to stimulate autologous allergen-specific T cell lines, we show that AEC-imprinted DCs inhibit T cell proliferation...... production of both Th1 and Th2 cytokines upon re-challenge. The inhibitory effects of AECs' contact with DCs were absent when allergen extract-loaded DCs had only been exposed to AECs supernatants, but present after direct contact with AECs. We conclude that direct contact between DCs and AECs inhibits T...

Current challenges of Germany’s energy transition project and competing strategies of challengers and incumbents: The case of direct marketing of electricity from renewable energy sources

International Nuclear Information System (INIS)

Wassermann, Sandra; Reeg, Matthias; Nienhaus, Kristina

2015-01-01

Electricity generated by renewable energies (RES-E) already accounts for 25% of Germany’s electricity supply. This has led to recent discussions for a better market integration of RES-E. The paper examines how competing actors and their ideas on market integration developed new services for direct marketing according to their respective origins and tried to shape the regulatory framework. The paper analyses this process and explains the current shape of the field of direct marketing. Medium-sized structured actors, who favoured RES-E integration via the conventional wholesale power markets, and who formed early close coalitions with RES-E power producers at the same time, have been most successful in terms of market shares. Moreover, they have been very successful for different reasons in building-up coalitions with governance units and influencing the field rules and routines. Based on those findings, the paper will conclude with some policy advices for the future adjustment of the current regulative frameworks. As long as there is no evidence of how RES-E can be integrated most effectively and efficiently, policies should maintain a competition between different direct marketing strategies to find out which strategies serve the best in terms of achieving a successful energy transition. - Highlights: • Innovation sociological analysis of the market integration of electricity from renewables in the German electricity markets. • Direct marketing of RES-E seen as a new strategic action field in the German “Energiewende”. • Strategies of incumbent and challenger actors to shape the rules of the field. • Suggestions for the future design of policy instruments for direct marketing of RES-E

Planning for the Future of Geo-Cybereducation: Outcomes of the Workshop, Challenges, and Future Directions

Science.gov (United States)

Ryan, J. G.; Eriksson, S. C.

2010-12-01

Inspired by the recommendations of the NSF report “Fostering Learning in the Networked World: The Cyberlearning Opportunity and Challenge” (NSF08204), the NSF National STEM Digital Learning program funded “Planning for the Future of Geocybereducation” Workshop sought to bring together leaders from the geoscience education community, from major geoscience research initiatives, and from the growing public- and private-sector geoscience information community. The objectives of the workshop were to begin conversations aimed at identifying best practices and tools for geoscience cyber-education, in the context of both the changing nature of learners and of rapidly evolving geo-information platforms, and to provide guidance to the NSF as to necessary future directions and needs for funding. 65 participants met and interacted live for the two-day workshop, with ongoing post-meeting virtual interactions via a collaborative workspace (www.geocybered.ning.com). Topics addressed included the rapidly changing character of learners, the growing capabilities of geoscience information systems and their affiliated tools, and effective models for collaboration among educators, researchers and geoinformation specialists. Discussions at the meeting focused on the implications of changing learners on the educational process, the challenges for teachers and administrators in keeping pace, and on the challenges of communication among these divergent professional communities. Ongoing virtual discussions and collaborations have produced a draft workshop document, and the workshop conveners are maintaining the workshop site as a venue for ongoing discussion and interaction. Several key challenges were evident from the workshop discussions and subsequent interactions: a) the development of most of the large geoinformatics and geoscience research efforts were not pursued with education as a significant objective, resulting in limited financial support for such activities after the

Global surgery for pediatric hydrocephalus in the developing world: a review of the history, challenges, and future directions.

Science.gov (United States)

Muir, Ryan T; Wang, Shelly; Warf, Benjamin C

2016-11-01

OBJECTIVE Pediatric hydrocephalus is one of the most common neurosurgical conditions and is a major contributor to the global burden of surgically treatable diseases. Significant health disparities exist for the treatment of hydrocephalus in developing nations due to a combination of medical, environmental, and socioeconomic factors. This review aims to provide the international neurosurgery community with an overview of the current challenges and future directions of neurosurgical care for children with hydrocephalus in low-income countries. METHODS The authors conducted a literature review around the topic of pediatric hydrocephalus in the context of global surgery, the unique challenges to creating access to care in low-income countries, and current international efforts to address the problem. RESULTS Developing countries face the greatest burden of pediatric hydrocephalus due to high birth rates and greater risk of neonatal infections. This burden is related to more general global health challenges, including malnutrition, infectious diseases, maternal and perinatal risk factors, and education gaps. Unique challenges pertaining to the treatment of hydrocephalus in the developing world include a preponderance of postinfectious hydrocephalus, limited resources, and restricted access to neurosurgical care. In the 21st century, several organizations have established programs that provide hydrocephalus treatment and neurosurgical training in Africa, Central and South America, Haiti, and Southeast Asia. These international efforts have employed various models to achieve the goals of providing safe, sustainable, and cost-effective treatment. CONCLUSIONS Broader commitment from the pediatric neurosurgery community, increased funding, public education, surgeon training, and ongoing surgical innovation will be needed to meaningfully address the global burden of untreated hydrocephalus.

DIRECTIONS AND CHALLENGES IN GLOBAL AGRICULTURE

Directory of Open Access Journals (Sweden)

Daniela POPA

2013-01-01

Full Text Available The development of global agricultural market has been at the forefront of professional studies. Expert opinions have quite differing views as to whether the world’s food production will be able to supply the huge demand of growing population. This scientific paper provides a general overview of global agricultural directions, including views on whether agricultural productivity increases will be able to keep with food demand increases and price trends. The scientific paper has focused on the present state of the agricultural market and on the analysis of the key factors defining the tasks of the agricultural sector in the near future, with a special attention to the case of Republic of Moldova.

NANOG Is Multiply Phosphorylated and Directly Modified by ERK2 and CDK1 In Vitro

Directory of Open Access Journals (Sweden)

Justin Brumbaugh

2014-01-01

Full Text Available NANOG is a divergent homeobox protein and a core component of the transcriptional circuitry that sustains pluripotency and self-renewal. Although NANOG has been extensively studied on the transcriptional level, little is known regarding its posttranslational regulation, likely due to its low abundance and challenging physical properties. Here, we identify eleven phosphorylation sites on endogenous human NANOG, nine of which mapped to single amino acids. To screen for the signaling molecules that impart these modifications, we developed the multiplexed assay for kinase specificity (MAKS. MAKS simultaneously tests activity for up to ten kinases while directly identifying the substrate and exact site of phosphorylation. Using MAKS, we discovered site-specific phosphorylation by ERK2 and CDK1/CyclinA2, providing a putative link between key signaling pathways and NANOG.

The European water framework directive: A challenge for nearshore, coastal and continental shelf research

Science.gov (United States)

Borja, Ángel

2005-09-01

The European Water Framework Directive (WFD) establishes a framework for the protection of groundwater, inland surface waters, estuarine waters, and coastal waters. The WFD constitutes a new view of the water resources management in Europe because, for the first time, water management is: (i) based mainly upon biological and ecological elements, with ecosystems being at the centre of the management decisions; (ii) applied to European water bodies, as a whole; and (iii) based upon the whole river basin, including also the adjacent coastal area. Although the marine water bodies affected by the WFD relate to only 19.8% of the whole of the European continental shelf, its application constitutes a challenge and an opportunity in nearshore, coastal and continental shelf research. This contribution highlights some of the main tasks and the research to be undertaken in the coming years, proposing investigations into: typologies; physico-chemical processes; indicator species; reference conditions; integration of the quality assessment; methodologies in determining ecological status, etc.

Direct human DNA protection by Coriolus versicolor (Yunzhi) extract.

Science.gov (United States)

Szeto, Yim Tong; Lau, Po Chun; Kalle, Wouter; Pak, Sok Cheon

2013-07-01

Scientific evidence has shown Coriolus versicolor (L. ex Fr.) Quel (also known as Yunzhi) has the role of immunomodulator in therapeutic effect. The aim of this in vitro study was to investigate the antioxidative effect of Yunzhi and to explore the mechanisms behind its DNA protection. Commercial Yunzhi extract was dissolved in water and diluted in five concentrations (10(1)-10(5) μg/L) with appropriate buffers. Lymphocytes harvested from three healthy subjects were incubated with Yunzhi extract for 30 min. Cells were then subjected to 5 min oxidant challenge by 45 μM hydrogen peroxide. The standard alkaline comet (SAC) assay and lysed cell comet (LCC) assay were performed in parallel. DNA damage of each treatment was scored under a fluorescence microscope and compared with the cells without Yunzhi pretreatment. U-shaped dose-response was seen in both versions of the comet assay. Yunzhi at 10(4) μg/L demonstrated a genoprotective effect against oxidative damage in the SAC assay (25% decrease in comet score). In the LCC assay, a trend of protection in lymphocytes was observed but it did not reach statistical significance. A direct antioxidant effect of Yunzhi against oxidant challenge on the DNA of lymphocytes was evidenced. The active component in Yunzhi was likely to be membrane permeable.

A novel co-processed directly compressible release-retarding polymer: In vitro, solid state and in vivo evaluation

Directory of Open Access Journals (Sweden)

Prashant Kumar Choudhari

2018-06-01

Full Text Available Directly compressible (DC co-processed excipient capable of providing nearly zero order release with improved functionality was developed without any chemical modification by employed various techniques such as physical mixing, high shear mixer granulation and spray drying. Co-processed excipient was developed by using release retarding polymer Eudragit RSPO, separately, in combination with different concentration of hydroxyl propyl methyl cellulose 100 cps (Methocel K100 LV, HPMC, ethyl cellulose (Ethocel N50, EC and hydroxyl propyl cellulose (Klucel EF, HPC. All co-processed excipients were evaluated for their flow properties in terms of angle of repose, bulk density, tapped density, compressibility index and Hausner's ratio. Out of eighteen combinations, the nine co-processed excipients exhibited promising flow properties were found suitable for direct compression and formulated as tablets. Metoprolol succinate, a BCS Class I drug, was selected as a model drug and the formulation was developed employing direct compression approach. The developed tablets were evaluated for physical parameters like uniformity of weight, thickness, hardness, friability and assay. In vitro dissolution study confirms that formulation prepared using co-processed excipient showed sustained drug release. The optimized tablet formulation was characterized by DSC, FTIR and PXRD which confirms the absence of any chemical change during co-processing. The optimized formulation was kept for stability study for six months as per ICH guidelines and found to be stable. In vivo pharmacokinetic study of optimized formulation in rats showed similar pharmacokinetic behaviour as was observed with the marketed brand. Study revealed that co-processed excipient has advantage over polymers with single property and can be utilised for sustained release formulation. Keywords: Co-processed excipient, Metoprolol succinate, Extended-release, Direct compression, Zero-order release

Modeling Acute Traumatic Hemorrhagic Shock Injury: Challenges and Guidelines for Preclinical Studies.

Science.gov (United States)

Tremoleda, Jordi L; Watts, Sarah A; Reynolds, Penny S; Thiemermann, Christoph; Brohi, Karim

2017-12-01

Trauma is responsible for a large proportion of the world's burden of disease, and is by far the biggest killer of young adults. Hemorrhage is the leading cause of preventable death and its effects are directly correlated with the incidence multi-organ failure in survivors. Trauma research is challenging due to patient heterogeneity, limited randomized controlled trials, and in vitro studies that fail to mimic the systemic injury response. Preclinical research remains essential for mechanistic and therapeutic discovery. Yet modeling the multifaceted nature of traumatic injury poses important experimental and welfare challenges associated with the onset of injury and prehospital and intra-operative care, the limited inter-species validation of coagulation profiles, the use of anesthesia/analgesia, and its impact on the systemic response to trauma; and the challenge of sustaining intensive care in recovery models. Proper model selection depends on the purpose of a given model and the criteria by which the experimental readouts will be clinically relevant. Such complexity warrants further refinement of experimental methodology and outcome measures to improve its clinical efficacy, while ensuring animal well-being. We review the experimental methodologies currently used for modeling traumatic hemorrhagic shock and addressing their impact on clinical translation. The aim of the review is to improve transparency and form a consensus when reporting methodology in trauma modeling.

In vitro Alternative Methodologies for Central Nervous System Assessment: A Critique using Nanoscale Materials as an Example.

Science.gov (United States)

Identifying the potential health hazards to the central nervous system of a new family of materials presents many challenges. Whole-animal toxicity testing has been the tradition, but in vitro methods have been steadily gaining popularity. There are numerous challenges in testing...

Tolerance induction to a thymus-dependent antigen in vitro: treatment of nonadherent cells with tolerogen biologically filtered in vitro

International Nuclear Information System (INIS)

Geyer, J.W.; Kong, Y.M.

1974-01-01

Highly tolerogenic bovine gamma globulin (BGG), a thymus-dependent antigen, was prepared by biologic filtration in vitro. It readily induced tolerance in vivo in BALB/c mice and also rendered their nonadherent lymph node cells tolerant after in vitro incubation. Biologic filtration in vitro was carried out by incubating 2.5 x 10 7 lymph node cells with 10 mg of nontolerogenic BGG in 10 ml of Eagle's medium containing 2 percent normal mouse serum at 37 0 C for 6 hr. The BGG-containing medium was clarified by centrifugation and was used without further dilution. For tolerance induction in vitro, lymph node cells were separated into adherent and nonadherent populations on Falcon plastic. These cells were incubated for 0 to 18 hr at 37 0 C with biologically filtered BGG (bBGG). After incubation, the cells were washed three times and (2 to 2.5) x 10 7 nonadherent or 4 x 10 6 adherent cells were injected iv with their untreated counterpart into lethally irradiated mice which had received 10 6 bone marrow cells. The recipients were then challenged with 300 μg of aggregated BGG, and tolerance was assayed by the elimination of labeled BGG, rosette formation, and passive hemagglutination. Spleen cells were similarly treated for comparison. Our findings show that tolerance was not induced in vitro in adherent lymph node cells. However, in the nonadherent populations, those from the lymph node but not the spleen were rendered tolerant. The acquisition of tolerance in vitro was gradual. It was dependent upon the length of exposure to bBGG and required at least 6 hr

Is in vitro meat the solution for the future?

Science.gov (United States)

Hocquette, Jean-François

2016-10-01

The production of in vitro meat regularly generates media interest because of the contribution it could, at first glance, make to the issue of feeding humankind while also protecting the environment and respecting animals. However, the majority of experts considers that there are still numerous technological obstacles that have to be overcome to produce in vitro meat. In addition, even if in vitro meat could eliminate the supposed lack of well-being of livestock and has the potential to free up cultivable land, other supposed advantages are questionable and not always agreed upon by the scientific community. However, another major problem for the commercialisation of in vitro meat would be its acceptance by consumers, even if some consumers are ready to taste it at least once. In particular, the artificial nature of the product goes against the growing demand for natural products in many countries. The consumption of in vitro meat will depend on a conflict of values at an individual or collective level. The reality is that a range of other complementary solutions already exist which meet the challenges of food supply in our society, but which are less saleable to the media. Copyright © 2016 Elsevier Ltd. All rights reserved.

Engineering kidney cells: reprogramming and directed differentiation to renal tissues.

Science.gov (United States)

Kaminski, Michael M; Tosic, Jelena; Pichler, Roman; Arnold, Sebastian J; Lienkamp, Soeren S

2017-07-01

Growing knowledge of how cell identity is determined at the molecular level has enabled the generation of diverse tissue types, including renal cells from pluripotent or somatic cells. Recently, several in vitro protocols involving either directed differentiation or transcription-factor-based reprogramming to kidney cells have been established. Embryonic stem cells or induced pluripotent stem cells can be guided towards a kidney fate by exposing them to combinations of growth factors or small molecules. Here, renal development is recapitulated in vitro resulting in kidney cells or organoids that show striking similarities to mammalian embryonic nephrons. In addition, culture conditions are also defined that allow the expansion of renal progenitor cells in vitro. Another route towards the generation of kidney cells is direct reprogramming. Key transcription factors are used to directly impose renal cell identity on somatic cells, thus circumventing the pluripotent stage. This complementary approach to stem-cell-based differentiation has been demonstrated to generate renal tubule cells and nephron progenitors. In-vitro-generated renal cells offer new opportunities for modelling inherited and acquired renal diseases on a patient-specific genetic background. These cells represent a potential source for developing novel models for kidney diseases, drug screening and nephrotoxicity testing and might represent the first steps towards kidney cell replacement therapies. In this review, we summarize current approaches for the generation of renal cells in vitro and discuss the advantages of each approach and their potential applications.

Bacteriocinogenic LAB Strains for Fermented Meat Preservation: Perspectives, Challenges, and Limitations.

Science.gov (United States)

Favaro, Lorenzo; Todorov, Svetoslav Dimitrov

2017-12-01

Over the last decades, much research has focused on lactic acid bacteria (LAB) bacteriocins because of their potential as biopreservatives and their action against the growth of spoilage microbes. Meat and fermented meat products are prone to microbial contamination, causing health risks, as well as economic losses in the meat industry. The use of bacteriocin-producing LAB starter or protective cultures is suitable for fermented meats. However, although bacteriocins can be produced during meat processing, their levels are usually much lower than those achieved during in vitro fermentations under optimal environmental conditions. Thus, the direct addition of a bacteriocin food additive would be desirable. Moreover, safety and technological characteristics of the bacteriocinogenic LAB must be considered before their widespread applications. This review describes the perspectives and challenges toward the complete disclosure of new bacteriocins as effective preservatives in the production of safe and "healthy" fermented meat products.

Data Science Meets the Clinician: Challenges and Future Directions.

Science.gov (United States)

Charitos, Efstratios I; Wilbring, Manuel; Treede, Hendrik

2018-01-01

In the last three decades a profound transformation of the medical profession has taken place. The modern clinician is required to consume vast amounts of information from clinical studies, critically reviewing evidence that may or may not lead to changes in clinical practice. The present article presents some challenges that this era of information poses to clinicians and patients. Georg Thieme Verlag KG Stuttgart · New York.

T Cell Genesis: In Vitro Veritas Est?

OpenAIRE

Brauer, Patrick M.; Singh, Jastaranpreet; Xhiku, Sintia; Zúñiga-Pflücker, Juan Carlos

2016-01-01

T cells, as orchestrators of the adaptive immune response, serve important physiological and potentially therapeutic roles, for example in cancer immunotherapy. T cells are readily isolated from patients; however, the yield of antigen-specific T cells is limited, thus making their clinical use challenging. Therefore, the generation of T lymphocytes from hematopoietic stem/progenitor cells (HSPCs) and human pluripotent stem cells (PSCs) in vitro provides an attractive method for large-scale pr...

Readout technologies for directional WIMP Dark Matter detection

International Nuclear Information System (INIS)

Battat, J.B.R.; Irastorza, I.G.; Aleksandrov, A.; Asada, T.; Baracchini, E.; Billard, J.; Bosson, G.; Bourrion, O.; Bouvier, J.; Buonaura, A.; Burdge, K.; Cebrián, S.

2016-01-01

The measurement of the direction of WIMP-induced nuclear recoils is a compelling but technologically challenging strategy to provide an unambiguous signature of the detection of Galactic dark matter. Most directional detectors aim to reconstruct the dark-matter-induced nuclear recoil tracks, either in gas or solid targets. The main challenge with directional detection is the need for high spatial resolution over large volumes, which puts strong requirements on the readout technologies. In this paper we review the various detector readout technologies used by directional detectors. In particular, we summarize the challenges, advantages and drawbacks of each approach, and discuss future prospects for these technologies.

Modeling of biomass-to-energy supply chain operations: Applications, challenges and research directions

International Nuclear Information System (INIS)

Mafakheri, Fereshteh; Nasiri, Fuzhan

2014-01-01

Reducing dependency on fossil fuels and mitigating their environmental impacts are among the most promising aspects of utilizing renewable energy sources. The availability of various biomass resources has made it an appealing source of renewable energy. Given the variability of supply and sources of biomass, supply chains play an important role in the efficient provisioning of biomass resources for energy production. This paper provides a comprehensive review and classification of the excising literature in modeling of biomass supply chain operations while linking them to the wider strategic challenges and issues with the design, planning and management of biomass supply chains. On that basis, we will present an analysis of the existing gaps and the potential future directions for research in modeling of biomass supply chain operations. - Highlights: • An extensive review of biomass supply chain operations management models presented in the literature is provided. • The models are classified in line with biomass supply chain activities from harvesting to conversion. • The issues surrounding biomass supply chains are investigated manifesting the need to novel modeling approaches. • Our gap analysis has identified a number of existing shortcomings and opportunities for future research

Plerocercoid growth factor (PGF), a human growth hormone (hGH) analogue produced by the tapeworm Spirometra mansonoides, has direct insulin-like action in adipose tissue of normal rats in vitro

International Nuclear Information System (INIS)

Salem, M.A.M.; Phares, C.K.

1986-01-01

The metabolic actions of GH can be divided into acute (insulin-like) and chronic (lipolytic/anti-insulin). The insulin-like actions of GH are most readily elicited in GH-deficient animals as GH induces resistance to its own insulin-like action. Like GH, PGF stimulates growth and cross-reacts with anti-hGH antibodies. Independent experiments were conducted comparing the direct actions of PGF to insulin or hGH in vitro. Insulin-like effects were determined by the ability of PGF, insulin or hGH to stimulate [U- 14 C]glucose metabolism in epidydimal fat pads from normal rats and by inhibition of epinephrine-stimulated lipolysis. Direct stimulation of lipolysis was used as anti-insulin activity. To determine if PGF competes for insulin or GH receptors, adipocytes (3 x 10 5 cells/ml) were incubated with either [ 125 I]insulin or [ 125 I]hGH +/- PGF, +/- insulin or +/- hGH. PGF stimulated glucose oxidation and 14 C-incorporation into lipids. Insulin, hGH and PGF inhibited lipolysis (33%, 29% and 34%, respectively). Adipose tissue was very sensitive to the lipolytic effect of hGH but PGF was neither lipolytic nor did it confer refractoriness to its insulin-like action. PGF bound to GH but not to insulin receptors. Therefore, PGF had direct insulin-like effects but did not stimulate lipolysis in tissue from normal rats in vitro

Surface functionalization of 3D glass-ceramic porous scaffolds for enhanced mineralization in vitro

Science.gov (United States)

Ferraris, Sara; Vitale-Brovarone, Chiara; Bretcanu, Oana; Cassinelli, Clara; Vernè, Enrica

2013-04-01

Bone reconstruction after tissue loosening due to traumatic, pathological or surgical causes is in increasing demand. 3D scaffolds are a widely studied solution for supporting new bone growth. Bioactive glass-ceramic porous materials can offer a three-dimensional structure that is able to chemically bond to bone. The ability to surface modify these devices by grafting biologically active molecules represents a challenge, with the aim of stimulating physiological bone regeneration with both inorganic and organic signals. In this research work glass ceramic scaffolds with very high mechanical properties and moderate bioactivity have been functionalized with the enzyme alkaline phosphatase (ALP). The material surface was activated in order to expose hydroxyl groups. The activated surface was further grafted with ALP both via silanization and also via direct grafting to the surface active hydroxyl groups. Enzymatic activity of grafted samples were measured by means of UV-vis spectroscopy before and after ultrasonic washing in TRIS-HCl buffer solution. In vitro inorganic bioactivity was investigated by soaking the scaffolds after the different steps of functionalization in a simulated body fluid (SBF). SEM observations allowed the monitoring of the scaffold morphology and surface chemical composition after soaking in SBF. The presence of ALP enhanced the in vitro inorganic bioactivity of the tested material.

Direct coercion – challenges for nursing practice

Directory of Open Access Journals (Sweden)

Karcz Edyta

2017-06-01

Full Text Available Introduction. Direct coercion is a part of medical intervention in which the patient’s autonomy as a basic human right is limited. In clinical practice, there are situations in which the use of coercion is indeed necessary to protect the patient and his/her environment against the effects of growing aggression. A very important element of action in such cases is to ensure the safety of both the patient through professional care, and of the personel executing the coercive measure.

Water management: Current and future challenges and research directions

Science.gov (United States)

Cosgrove, William J.; Loucks, Daniel P.

2015-06-01

Water distinguishes our planet compared to all the others we know about. While the global supply of available freshwater is more than adequate to meet all current and foreseeable water demands, its spatial and temporal distributions are not. There are many regions where our freshwater resources are inadequate to meet domestic, economic development and environmental needs. In such regions, the lack of adequate clean water to meet human drinking water and sanitation needs is indeed a constraint on human health and productivity and hence on economic development as well as on the maintenance of a clean environment and healthy ecosystems. All of us involved in research must find ways to remove these constraints. We face multiple challenges in doing that, especially given a changing and uncertain future climate, and a rapidly growing population that is driving increased social and economic development, globalization, and urbanization. How best to meet these challenges requires research in all aspects of water management. Since 1965, the journal Water Resources Research has played an important role in reporting and disseminating current research related to managing the quantity and quality and cost of this resource. This paper identifies the issues facing water managers today and future research needed to better inform those who strive to create a more sustainable and desirable future.

In vitro function of the aryl hydrocarbon receptor predicts in ...

Science.gov (United States)

Differences in sensitivity to dioxin-like compounds (DLCs) among species and taxa presents a major challenge to ecological risk assessments. Activation of the aryl hydrocarbon receptor (AHR) regulates adverse effects associated with exposure to DLCs in vertebrates. Prior investigations demonstrated that sensitivity to activation of the AHR1 (50% effect concentration; EC50) in an in vitro luciferase reporter gene (LRG) assay was predictive of the sensitivity of embryos (lethal dose to cause 50% lethality; LD50) across all species of birds for all DLCs. However, nothing was known about whether sensitivity to activation of the AHR is predictive of sensitivity of embryos of fishes to DLCs. Therefore, this study investigated in vitro sensitivities of AHR1s and AHR2s to the model DLC, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), among eight species of fish of known sensitivities of embryos to TCDD. AHR1s and AHR2s of all fishes were activated by TCDD in vitro. There was no significant linear relationship between in vitro sensitivity of AHR1 and in vivo sensitivity among the investigated fishes (R2 = 0.33, p = 0.23). However, there was a significant linear relationship between in vitro sensitivity of AHR2 and in vivo sensitivity among the investigated fishes (R2 = 0.97, p = fishes was compared to the previously generated linear relationship between in vitro s

Useful predictors of ovarian stimulation response in women undergoing in vitro fertilization

NARCIS (Netherlands)

van der Stege, JG; van der Linden, PJQ

2001-01-01

Eighty-seven patients undergoing in vitro fertilization were studied to evaluate the prognostic value of different tests in predicting ovarian stimulation response. We studied basal follicle-stimulating hormone (FSH) value on cycle day 3, the clomiphene citrate (CC) challenge test and serum

Hydrogeologic characterization of the former Vitro processing site, Salt Lake City, Utah

International Nuclear Information System (INIS)

1993-01-01

During fiscal year (FY) 1992, the US Department of Energy received Congressional direction to investigate whether contamination from former processing activities is present in groundwater and soils at the former Vitro processing site in Salt Lake City, Utah. A total of $100,000 was appropriated for this activity. The surface of the Vitro site was cleaned up by the state of Utah under the Uranium Mill Tailings Remedial Action (UMTRA) Project during the mid- to late 1980s. The basis for the directive was the desire of the site owners, the Central Valley Water Reclamation Facility (CVWRF), to develop the site. This report, as well as the final Vitro Site Certification Report and the Clive Completion Report (both to be prepared and submitted by the state of Utah), will have to be reviewed and concurred with by the US Nuclear Regulatory Commission (NRC) prior to release of the Vitro site for restricted development. The groundwater and soil investigation was performed by the Technical Assistance Contractor (TAC) to the DOE UMTRA Project Office during FY92. The investigation at the Vitro site consisted of the installation of monitoring wells and soil borings, aquifer testing, and the collection and analyses of groundwater and soil samples. This report presents the results of this hydrogeologic investigation

Direct Cell-Cell Contact between Mesenchymal Stem Cells and Endothelial Progenitor Cells Induces a Pericyte-Like Phenotype In Vitro

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Markus Loibl

2014-01-01

Full Text Available Tissue engineering techniques for the regeneration of large bone defects require sufficient vascularisation of the applied constructs to ensure a sufficient supply of oxygen and nutrients. In our previous work, prevascularised 3D scaffolds have been successfully established by coculture of bone marrow derived stem cells (MSCs and endothelial progenitor cells (EPCs. We identified stabilising pericytes (PCs as part of newly formed capillary-like structures. In the present study, we report preliminary data on the interactions between MSCs and EPCs, leading to the differentiation of pericyte-like cells. MSCs and EPCs were seeded in transwell cultures, direct cocultures, and single cultures. Cells were cultured for 10 days in IMDM 10% FCS or IMDM 5% FCS 5% platelet lysate medium. Gene expression of PC markers, CD146, NG2, αSMA, and PDGFR-β, was analysed using RT-PCR at days 0, 3, 7, and 10. The upregulation of CD146, NG2, and αSMA in MSCs in direct coculture with EPCs advocates the MSCs’ differentiation towards a pericyte-like phenotype in vitro. These results suggest that pericyte-like cells derive from MSCs and that cell-cell contact with EPCs is an important factor for this differentiation process. These findings emphasise the concept of coculture strategies to promote angiogenesis for cell-based tissue engineered bone grafts.

Flex-Fuel Two-Stroke Snowmobile: Development of a Flex-Fuel, Two-Stroke, Direct-Injection Snowmobile for Use in the Clean Snowmobile Challenge and National Parks

Science.gov (United States)

2009-09-01

The University of Idaho's entry into the 2009 SAE Clean Snowmobile Challenge (CSC) was a semi-direct-injection (SDI) two-stroke powered REV-XP snowmobile modified to use flex fuel. The flex fuel engine produces stock engine power on any blend of etha...

In vitro meat production system: why and how?

Science.gov (United States)

Sharma, Shruti; Thind, Sukhcharanjit Singh; Kaur, Amarjeet

2015-12-01

Due to the nutritional importance and the sustained popularity of meat as a foodstuff, the livestock production sector has been expanding incessantly. This exponential growth of livestock meat sector poses a gigantic challenge to the sustainability of food production system. A new technological breakthrough is being contemplated to develop a substitute for livestock meat. The idea is to grow meat in a culture in the lab and manipulate its composition selectively. This paper aims to discuss the concept of In Vitro Meat production system, articulate the underlying technology and analyse the context of its implications, as proposed by several scientists and stakeholders. The challenges facing this emerging technology have also been discussed.

A Salmonella typhimurium ghost vaccine induces cytokine expression in vitro and immune responses in vivo and protects rats against homologous and heterologous challenges.

Directory of Open Access Journals (Sweden)

Nagarajan Vinod

Full Text Available Salmonella enteritidis and Salmonella typhimurium are important food-borne bacterial pathogens, which are responsible for diarrhea and gastroenteritis in humans and animals. In this study, S. typhimurium bacterial ghost (STG was generated based on minimum inhibitory concentration (MIC of sodium hydroxide (NaOH. Experimental studies performed using in vitro and in vivo experimental model systems to characterize effects of STG as a vaccine candidate. When compared with murine macrophages (RAW 264.7 exposed to PBS buffer (98.1%, the macrophages exposed to formalin-killed inactivated cells (FKC, live wild-type bacterial cells and NaOH-induced STG at 1 × 108 CFU/mL showed 85.6%, 66.5% and 84.6% cell viability, respectively. It suggests that STG significantly reduces the cytotoxic effect of wild-type bacterial cells. Furthermore, STG is an excellent inducer for mRNAs of pro-inflammatory cytokine (TNF-α, IL-1β and factor (iNOS, anti-inflammatory cytokine (IL-10 and dual activities (IL-6 in the stimulated macrophage cells. In vivo, STG vaccine induced humoral and cellular immune responses and protection against homologous and heterologous challenges in rats. Furthermore, the immunogenicity and protective efficacy of STG vaccine were compared with those of FKC and non-vaccinated PBS control groups. The vaccinated rats from STG group exhibited higher levels of serum IgG antibody responses, serum bactericidal antibodies, and CD4+ and CD8+ T-cell populations than those of the FKC and PBS control groups. Most importantly, after challenge with homologous and heterologous strains, the bacterial loads in the STG group were markedly lower than the FKC and PBS control groups. In conclusion, these findings suggest that the STG vaccine induces protective immunity against homologous and heterologous challenges.

Optimization of the Contrast Mixture Ratio for Simultaneous Direct MR and CT Arthrography: an in Vitro Study

International Nuclear Information System (INIS)

Choi, Ja Young; Hong, Sung Hwan; Kim, Na Ra; Jun, Woo Sun; Moon, Sung Gyu; Kang, Heung Sik; Lee, Joon Woo; Choi, Jung Ah

2008-01-01

This study was designed to determine the optimal mixture ratio of gadolinium and iodinated contrast agent for simultaneous direct MR arthrography and CT arthrography. An in vitro study was performed utilizing mixtures of gadolinium at six different concentrations (0.625, 1.25, 2.5, 5.0, 10 and 20 mmol/L) and iodinated contrast agent at seven different concentrations (0, 12.5, 25, 37.5, 50, 75 and 92-99.9%). These mixtures were placed in tissue culture plates, and were then imaged with CT and MR (with T1-weighted sequences, proton-density sequences and T2-weighted sequences). CT numbers and signal intensities were measured. Pearson's correlation coefficients were used to assess the correlations between the gadolinium/iodinated contrast agent mixtures and the CT numbers/MR signal intensities. Scatter diagrams were plotted for all gadolinium/iodinated contrast agent combinations and two radiologists in consensus identified the mixtures that yielded the optimal CT numbers and MR signal intensities. The CT numbers showed significant correlation with iodinated contrast concentrations (r = 0.976, p < 0.001), whereas the signal intensities as measured on MR images showed a significant correlation with both gadolinium and iodinated contrast agent concentrations (r = -484 to -0.719, p < 0.001). A review of the CT and MR images, graphs, and scatter diagram of 42 combinations of the contrast agent showed that a concentration of 1.25 mmol/L gadolinium and 25% iodinated contrast agent was the best combination for simultaneous CT and MR imaging. A mixture of 1.25 mmol/L gadolinium and 25% iodinated contrast agent was found to be optimal for simultaneous direct MR arthrography and CT arthrography

Guiding tissue regeneration with ultrasound in vitro and in vivo

Science.gov (United States)

Dalecki, Diane; Comeau, Eric S.; Raeman, Carol H.; Child, Sally Z.; Hobbs, Laura; Hocking, Denise C.

2015-05-01

Developing new technologies that enable the repair or replacement of injured or diseased tissues is a major focus of regenerative medicine. This paper will discuss three ultrasound technologies under development in our laboratories to guide tissue regeneration both in vitro and in vivo. A critical obstacle in tissue engineering is the need for rapid and effective tissue vascularization strategies. To address this challenge, we are developing acoustic patterning techniques for microvascular tissue engineering. Acoustic radiation forces associated with ultrasound standing wave fields provide a rapid, non-invasive approach to spatially pattern cells in three dimensions without affecting cell viability. Acoustic patterning of endothelial cells leads to the rapid formation of microvascular networks throughout the volumes of three-dimensional hydrogels, and the morphology of the resultant microvessel networks can be controlled by design of the ultrasound field. A second technology under development uses ultrasound to noninvasively control the microstructure of collagen fibers within engineered tissues. The microstructure of extracellular matrix proteins provides signals that direct cell functions critical to tissue regeneration. Thus, controlling collagen microfiber structure with ultrasound provides a noninvasive approach to regulate the mechanical properties of biomaterials and control cellular responses. The third technology employs therapeutic ultrasound to enhance the healing of chronic wounds. Recent studies demonstrate increased granulation tissue thickness and collagen deposition in murine dermal wounds exposed to pulsed ultrasound. In summary, ultrasound technologies offer noninvasive approaches to control cell behaviors and extracellular matrix organization and thus hold great promise to advance tissue regeneration in vitro and in vivo.

In vitro models for the prediction of in vivo performance of oral dosage forms

DEFF Research Database (Denmark)

Kostewicz, Edmund S; Abrahamsson, Bertil; Brewster, Marcus

2014-01-01

Accurate prediction of the in vivo biopharmaceutical performance of oral drug formulations is critical to efficient drug development. Traditionally, in vitro evaluation of oral drug formulations has focused on disintegration and dissolution testing for quality control (QC) purposes. The connection...... of formulations that rely on complex intraluminal processes (e.g. solubilization, supersaturation, precipitation…) remains extremely challenging. Concomitantly, the increasing demand for complex formulations to overcome low drug solubility or to control drug release rates urges the development of new in vitro...

Short communication. N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU) enhances in vitro direct shoot organogenesis of Citrus aurantium L. epicotyl segments compared to other commonly used cytokinins

Energy Technology Data Exchange (ETDEWEB)

Roussos, P. A.; Dimitriou, G.; Voloudakis, A. E.

2011-07-01

The effect of three concentrations of five different cytokinins, i.e. 6-benzylamino purine, 2-isopentyl adenine, kinetin (Kin), thidiazuron and N-(2-chloro-4-pyridyl)-N-phenylurea (4-CPPU), was evaluated on the in vitro direct shoot organogenesis of epicotyl explants of sour orange (Citrus aurantium L.). The basal medium used was that of Murashige and Tucker and epicotyl explants were incubated in medium supplemented with the prementioned cytokinins for 45 days. The addition of Kin and 4-CPPU in the medium enhanced the direct shoot organogenesis of sour orange epicotyl segments. The concentration of each of these two cytokinins which gave the best results, was combined with indole-3-acetic acid (IAA) or {alpha}-naphthalene acetic acid ({alpha}-NAA) at concentrations ranging from 0.01 mg L{sup -}1 to 0.2 mg L{sup -}1. The inclusion of IAA at 0.2 mg L{sup -}1 in the medium with 4-CPPU at 0.05 mg L{sup -}1 resulted in 100% successful direct shoot organogenesis, while the combination of Kin at 0.25 mg L{sup -}1 with IAA or {alpha}-NAA each at 0.01 mg L{sup -}1 presented equally high organogenesis percentages (91.7%). The incubation of the produced shoots, in medium supplemented with either indole-3-butyric acid or {alpha}-NAA resulted in high rooting percentages (up to 90%) and the rooted explants were successfully acclimatized under mist (85%). Although 4-CPPU has been used in in vitro culture of various species, this is the first report on its use in the direct shoot organogenesis of citrus species and could be of great value in citrus genetic transformation protocols using epicotyl segments, since this cytokinin resulted in the absolute organogenesis percentage. (Author) 20 refs.

Directing Creativity

DEFF Research Database (Denmark)

Darsø, Lotte; Ibbotson, Piers

2008-01-01

In this article we argue that leaders facing complex challenges can learn from the arts, specifically that leaders can learn by examining how theatre directors direct creativity through creative constraints. We suggest that perceiving creativity as a boundary phenomenon is helpful for directing it....... Like leaders, who are caught in paradoxical situations where they have to manage production and logistics simultaneously with making space for creativity and innovation, theatre directors need to find the delicate balance between on one hand renewal of perceptions, acting and interaction...... and on the other hand getting ready for the opening night. We conclude that the art of directing creativity is linked to developing competencies of conscious presence, attention and vigilance, whereas the craft of directing creativity concerns communication, framing and choice....

Direct satellite TV - The 12-GHz challenge

Science.gov (United States)

Fawcette, J.

1982-02-01

Manufacturers in Japan and Europe are developing the hardware necessary for commercially feasible direct broadcast satellite TV, including high-frequency circuits and mini-dishes for spacecasting. US companies are lagging behind due to formidable regulatory and legal difficulties. The article focuses on efforts to develop simple, inexpensive receivers which will be able to convert 12-GHz satellite transmissions into high-quality TV images. Three basic receiver designs are being developed: the mixer-downcaster, microwave integrated circuits using FET-preamplifier front ends with transistors connected by bond-wires, and monolithic gallium arsenide integrated circuits. Several companies are on the verge of introducing commercialized receivers utilizing these different basic designs.

In vitro fertilization and artificial activation of eggs of the direct-developing anuran Eleutherodactylus coqui

Directory of Open Access Journals (Sweden)

Toro Esteban

2004-08-01

Full Text Available Abstract Although much is known about the reproductive biology of pond-breeding frogs, there is comparatively little information about terrestrial-breeding anurans, a highly successful and diverse group. This study investigates the activation and in vitro fertilization of eggs of the Puerto Rican coqui frog obtained by hormonally induced ovulation. We report that spontaneous activation occurs in 34% of eggs, probably in response to mechanical stress during oviposition. Artificial activation, as evidenced by the slow block to polyspermy and the onset of zygote division, was elicited both by mechanical stimulation and calcium ionophore exposure in 64% and 83% of the cases, respectively. Finally, one in vitro fertilization protocol showed a 27% success rate, despite the fact that about one third of all unfertilized eggs obtained by hormone injection auto-activate. We expect these findings to aid in the conservation effort of Eleutherodactylus frogs, the largest vertebrate genus.

"I'm just trying to cope for both of us": Challenges and supports of family caregivers in participant-directed programs.

Science.gov (United States)

Milliken, Aimee; Mahoney, Ellen K; Mahoney, Kevin J; Mignosa, Kate; Rodriguez, Isabella; Cuchetti, Catherine; Inoue, Megumi

2018-05-17

Recently, national attention has focused on the needs of family caregivers providing complex chronic care, noting the necessity to better understand the scope of challenges they encounter. Although a robust body of literature exists about the scope of family caregiving, little is known specifically about the experiences and perspectives of family caregivers who support participant directed (PD) participants, particularly across the caregiving trajectory. Therefore, the aim of this qualitative descriptive study was to describe what family caregivers of individuals with developmental disabilities, physical disabilities, aging, or chronic health conditions identify as the challenges they experience as complex, and their perceptions of the effectiveness and gaps in family support resources in PD. Semi-structured interviews were audio-recorded with a purposive sample of caregivers. Transcribed interviews were analyzed using conventional content analysis. Fifty-four caregivers of individuals with a range of disabilities participated (age 34-78, M 59.9 ± 8.8; male 19%; spouse 17%, parent 61%). Six categories emerged from the analysis: contextualizing complexity, complexity in transitions, coping with complexity: advocacy & isolation, supportive support, unsupportive support, and systems challenges. Caregivers emphasized the interplay between unpredictability, transitions, and complexity and the interaction between the person receiving support, the caregiver's own situation, and the environment. Findings highlight the need, and provide a guide, for family assessment and for tailoring interventions matched to the profiles and self-identified challenges of families living with disability. Social workers can learn what families see as complex and what support broker behaviors families find helpful, and which not.

Effects of Saccharomyces cerevisiae at direct addition or pre-incubation on in vitro gas production kinetics and degradability of four fibrous feeds

Directory of Open Access Journals (Sweden)

Mona M.Y. Elghandour

2014-04-01

Full Text Available The objective of this study was to evaluate the effects of Saccharomyces cerevisiae on in vitro gas production (GP kinetics and degradability of corn stover, oat straw, sugarcane bagasse and sorghum straw. Feedstuffs were incubated with different doses of yeast [0, 4, 8 and 12 mg/g dry matter (DM] at direct addition or 72 h pre-incubation. Rumen GP was recorded at 2, 4, 6, 8, 10, 12, 14, 24, 30, 48, 54 and 72 h of incubation. After 72 h, rumen pH and methane were determined and contents were filtrated for DM, neutral (NDF and acid detergent fibre (ADF degradability. Fibrous species×method of application×yeast interactions occurred (P<0.001 for all measured ruminal GP parameters and degradability. The direct addition or 72 h pre-incubation of S. cerevisiae with corn stover improved (P<0.05 GP and methane and decreased (P<0.05 the lag time (L and NDF degradability (NDFD. The direct addition of S. cerevisiae to oat straw increased (P<0.05 rate of GP (c and decreased (P<0.05 asymptotic GP (b. However, 72 h pre-incubation increased (P<0.05 c with linearly decreased b, DM degradability (DMD and NDFD. Applying S. cerevisiae for 72 h pre-incubation decreased (P<0.001 methane emission. The direct addition or 72 h pre-incubation of S. cerevisiae to sorghum straw increased (P<0.05 b, c, L, DMD and NDFD. Overall, the effect of dose varied among different feedstuffs and different application methods. Results suggested that the direct addition of S. cerevisiae could support and improve ruminal fermentation of lowquality forages at 4 to 12 g/kg DM.

Resource potential of bamboo, challenges and future directions towards sustainable management and utilization in Ethiopia

Directory of Open Access Journals (Sweden)

Getachew Desalegn

2014-08-01

Full Text Available Aim of study: Bamboo, the fastest growing and high yielding perennial plant of the world has more than 1500 species and 1500 versatile socio-economic uses and ecological services. Ethiopia has two indigenous bamboo species namely Yushania alpina and Oxytenantheria abyssinica, covering about one million ha with a wide distribution. The objective of this paper is to highlight the potential of bamboo resources, challenges including biodeterioration damage, opportunities and future research directions towards its sustainable management and rational utilization.Area of study: Bamboo resources of EthiopiaMaterial and Methods: Reconnaissance survey was done to some parts of the bamboo growing potential areas in Ethiopia besides the literature review. Main results: The bamboo resource, despite its socio-economic and environmental benefits, currently, in most areas has been under high pressure due to land use changes, bamboo mass- flowering, poor processing with low value addition, and damage by biodeteriorating agents (termites, beetles and fungi. The preservative tests on Ethiopian bamboos revealed low natural durability and highlighted the paramount importance of appropriate protection measures such as Tanalith and vehicles used motor oil to increase durability, service life and rational utilization of bamboo-based products and structures as potential alternative construction and furniture material.Research highlights: Therefore, integrated research and development interventions involving different propagation and managements techniques, harvesting season, processing, value addition including proper seasoning and preservation technologies and marketing are recommended to fill the information and technological gaps on sustainable management and rational utilization of this fast growing and multipurpose bamboo resources in Ethiopia.Key words: Bamboo; challenges; management; socio-economic and environmental significance; utilization.

Comparison of sensitizers in vitro

International Nuclear Information System (INIS)

Hall, E.J.; Astor, M.

1980-01-01

Clinical trials of misonidazole are already well underway, but no definitive conclusions are likely for several years. Meanwhile, attention in the laboratory is directed toward identifying or synthesizing alternative compounds, which equal misonidazole in radiosensitizing efficiency, while at the same time exhibiting less troublesome side effects. Two of the most promising contenders are desmethylmisonidazole and SR 2508. The present communication describes experiments with cells cultured in vitro designed to compare the properties of misonidazole with these two potential replacements

Redox cofactor engineering in industrial microorganisms: strategies, recent applications and future directions.

Science.gov (United States)

Liu, Jiaheng; Li, Huiling; Zhao, Guangrong; Caiyin, Qinggele; Qiao, Jianjun

2018-05-01

NAD and NADP, a pivotal class of cofactors, which function as essential electron donors or acceptors in all biological organisms, drive considerable catabolic and anabolic reactions. Furthermore, they play critical roles in maintaining intracellular redox homeostasis. However, many metabolic engineering efforts in industrial microorganisms towards modification or introduction of metabolic pathways, especially those involving consumption, generation or transformation of NAD/NADP, often induce fluctuations in redox state, which dramatically impede cellular metabolism, resulting in decreased growth performance and biosynthetic capacity. Here, we comprehensively review the cofactor engineering strategies for solving the problematic redox imbalance in metabolism modification, as well as their features, suitabilities and recent applications. Some representative examples of in vitro biocatalysis are also described. In addition, we briefly discuss how tools and methods from the field of synthetic biology can be applied for cofactor engineering. Finally, future directions and challenges for development of cofactor redox engineering are presented.

In vitro meat: A future animal-free harvest.

Science.gov (United States)

Bhat, Zuhaib Fayaz; Kumar, Sunil; Bhat, Hina Fayaz

2017-03-04

In vitro meat production is a novel idea of producing meat without involving animals with the help of tissue engineering techniques. This biofabrication of complex living products by using various bioengineering techniques is a potential solution to reduce the ill effects of current meat production systems and can dramatically transform traditional animal-based agriculture by inventing "animal-free" meat and meat products. Nutrition-related diseases, food-borne illnesses, resource use and pollution, and use of farm animals are some serious consequences associated with conventional meat production methods. This new way of animal-free meat production may offer health and environmental advantages by reducing environmental pollution and resource use associated with current meat production systems and will also ensure sustainable production of designer, chemically safe, and disease-free meat as the conditions in an in vitro meat production system are controllable and manipulatable. Theoretically, this system is believed to be efficient enough to supply the global demand for meat; however, establishment of a sustainable in vitro meat production would face considerably greater technical challenges and a great deal of research is still needed to establish this animal-free meat culturing system on an industrial scale.

Statistical Challenges in Modeling Big Brain Signals

KAUST Repository

Yu, Zhaoxia

2017-11-01

Brain signal data are inherently big: massive in amount, complex in structure, and high in dimensions. These characteristics impose great challenges for statistical inference and learning. Here we review several key challenges, discuss possible solutions, and highlight future research directions.

Centennial Challenges Program

Science.gov (United States)

Ortega, Sam; Eberly, Eric

2015-01-01

NASA's Centennial Challenges Program was initiated in 2005 to directly engage the public in the process of advanced technology development. The program offers incentive prizes to generate revolutionary solutions to problems of interest to NASA and the nation. The program seeks innovations from diverse and nontraditional sources. Competitors are not supported by government funding and awards are only made to successful teams when the challenges are met. In keeping with the spirit of the Wright Brothers and other American innovators, the Centennial Challenge prizes are offered to independent inventors including small businesses, student groups, and individuals. These independent inventors are sought to generate innovative solutions for technical problems of interest to NASA and the nation and to provide them with the opportunity to stimulate or create new business ventures.

Eccentric muscle challenge shows osteopontin polymorphism modulation of muscle damage.

Science.gov (United States)

Barfield, Whitney L; Uaesoontrachoon, Kitipong; Wu, Chung-Sheih; Lin, Stephen; Chen, Yue; Wang, Paul C; Kanaan, Yasmine; Bond, Vernon; Hoffman, Eric P

2014-08-01

A promoter polymorphism of the osteopontin (OPN) gene (rs28357094) has been associated with multiple inflammatory states, severity of Duchenne muscular dystrophy (DMD) and muscle size in healthy young adults. We sought to define the mechanism of action of the polymorphism, using allele-specific in vitro reporter assays in muscle cells, and a genotype-stratified intervention in healthy controls. In vitro reporter constructs showed the G allele to respond to estrogen treatment, whereas the T allele showed no transcriptional response. Young adult volunteers (n = 187) were enrolled into a baseline study, and subjects with specific rs28357094 genotypes enrolled into an eccentric muscle challenge intervention [n = 3 TT; n = 3 GG/GT (dominant inheritance model)]. Female volunteers carrying the G allele showed significantly greater inflammation and increased muscle volume change as determined by magnetic resonance imaging T1- and T2-weighted images after eccentric challenge, as well as greater decrement in biceps muscle force. Our data suggest a model where the G allele enables enhanced activities of upstream enhancer elements due to loss of Sp1 binding at the polymorphic site. This results in significantly greater expression of the pro-inflammatory OPN cytokine during tissue remodeling in response to challenge in G allele carriers, promoting muscle hypertrophy in normal females, but increased damage in DMD patients. © The Author 2014. Published by Oxford University Press.

Directional differentiation of chicken embryonic stem cells into ...

African Journals Online (AJOL)

Jane

2011-08-01

Aug 1, 2011 ... In this study, the differentiation potential of chicken ES cells was investigated ... Key words: Chicken embryonic stem cells, in vitro, directional differentiation, .... synthesized by using the Revert Aid first strand cDNA synthesis kit.

Formulation and in vitro evaluation of fast dissolving tablets of metoprolol tartrate

Directory of Open Access Journals (Sweden)

Mangesh Machhindranath Satpute

2013-12-01

Full Text Available The demand for fast dissolving tablets has been growing during the last decade, especially for elderly and children who have swallowing difficulties. In the present work, fast dissolving tablets of metoprolol tartrate, were prepared using sodium starch glycolate, sodium croscarmellose and crospovidone as superdisintegrants, by the direct compression method. The tablets prepared were evaluated for various parameters including weight variation, hardness, friability, in vitro dispersion time, drug-polymer interaction, drug content water absorption ratio, wetting time, in vitro drug release, FTIR and DSC studies. The tablets prepared by the direct compression method had a weight variation in the range of 145 mg to 152 mg, which is below ± 7.5%, a hardness of 3.6 kg/cm² to 4.5 kg/cm², percentage friability of 0.46% to 0.73%, in vitro dispersion time of 18 s to 125 s, drug content uniformity of between 98.12% and 100.03%, a water absorption ratio of 67% to 87%, wetting time of 32 sec. to 64 sec., and an in vitro drug release of 53.92% - 98.82% within 15 min. The IR spectral analysis and DSC study showed no drug interaction with formulation additives of the tablet, and the formulations indicated no significant changes in hardness, friability, drug content or in vitro drug release. Fast dissolving tablets of metoprolol tartrate have enhanced dissolution and will lead to improved bioavailability and more effective therapy.

Transfer of in vitro expanded T lymphocytes after activation with dendritomas prolonged survival of mice challenged with EL4 tumor cells.

Science.gov (United States)

Li, Jinhua; Theofanous, Leigh; Stickel, Sara; Bouton-Verville, Hilary; Burgin, Kelly E; Jakubchak, Susan; Wagner, Thomas E; Wei, Yanzhang

2007-07-01

Adoptive T cell transfer after in vitro expansion represents an attractive cancer immunotherapy. The majority of studies so far have been focusing on the expansion of tumor infiltrated lymphocytes (TIL) and some have shown very encouraging results. Recently, we have developed a unique tumor immune response activator, dendritomas, by fusion of dendritic cells and tumor cells. Animal studies and early clinical trials have shown that dendritomas are able to activate tumor specific immune responses. In this study, we hypothesized that naïve T cells can be primed with dendritomas and expanded in vitro to develop an adoptive transfer therapy for patients who do not have solid tumors, such as leukemia. T cells were isolated and purified from lymph nodes of mice. The cells were then incubated with dendritomas made from syngeneic DCs and tumor cells and expanded in vitro using Dynabeads mouse CD3/CD28 T cell expander for approximately three weeks. The in vitro primed and expanded T cells showed tumor cell specific CTL activity and increased secretion of IFN-gamma. Tumor bearing mice receiving the in vitro expanded T cells survived significantly longer than control mice. Furthermore, the depletion of regulator T cells enhanced the survival of the mice that received the adoptive transfer therapy.

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model.

Science.gov (United States)

Chang, Robert; Emami, Kamal; Wu, Honglu; Sun, Wei

2010-12-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

International Nuclear Information System (INIS)

Chang, Robert; Sun Wei; Emami, Kamal; Wu Honglu

2010-01-01

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically characterized for

Biofabrication of a three-dimensional liver micro-organ as an in vitro drug metabolism model

Energy Technology Data Exchange (ETDEWEB)

Chang, Robert; Sun Wei [Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, PA (United States); Emami, Kamal; Wu Honglu, E-mail: rcc34@drexel.ed, E-mail: sunwei@drexel.ed, E-mail: kamal.emami-1@nasa.go, E-mail: honglu.wu-1@nasa.go [Radiation Biophysics Laboratory, Human Adaptation and Countermeasures Office, NASA Johnson Space Center, Houston, TX (United States)

2010-12-15

In their normal in vivo matrix milieu, tissues assume complex well-organized three-dimensional architectures. Therefore, the primary aim in the tissue engineering design process is to fabricate an optimal analog of the in vivo scenario. This challenge can be addressed by applying emerging layered biofabrication approaches in which the precise configuration and composition of cells and bioactive matrix components can recapitulate the well-defined three-dimensional biomimetic microenvironments that promote cell-cell and cell-matrix interactions. Furthermore, the advent of and refinements in microfabricated systems can present physical and chemical cues to cells in a controllable and reproducible fashion unmatched with conventional cultures, resulting in the precise construction of engineered biomimetic microenvironments on the cellular length scale in geometries that are readily parallelized for high throughput in vitro models. As such, the convergence of layered solid freeform fabrication (SFF) technologies along with microfabrication techniques enables the creation of a three-dimensional micro-organ device to serve as an in vitro platform for cell culture, drug screening or to elicit further biological insights, particularly for NASA's interest in a flight-suitable high-fidelity microscale platform to study drug metabolism in space and planetary environments. The proposed model in this paper involves the combinatorial setup of an automated syringe-based, layered direct cell writing bioprinting process with micro-patterning techniques to fabricate a microscale in vitro device housing a chamber of bioprinted three-dimensional liver cell-encapsulated hydrogel-based tissue constructs in defined design patterns that biomimic the cell's natural microenvironment for enhanced biological functionality. In order to assess the structural formability and biological feasibility of such a micro-organ, reproducibly fabricated tissue constructs were biologically

5G backhaul challenges and emerging research directions: a survey

OpenAIRE

Jaber, Mona; Imran, Muhammad Ali; Tafazolli, Rahim; Tukmanov, Anvar

2016-01-01

5G is the next cellular generation and is expected to quench the growing thirst for taxing data rates and to enable the Internet of Things. Focused research and standardization work have been addressing the corresponding challenges from the radio perspective while employing advanced features, such as network densification, massive multiple-input-multiple-output antennae, coordinated multi-point processing, inter-cell interference mitigation techniques, carrier aggregation, and new spectrum ex...

Burnout After Patient Death: Challenges for Direct Care Workers.

Science.gov (United States)

Boerner, Kathrin; Gleason, Hayley; Jopp, Daniela S

2017-09-01

Direct care workers in long-term care can develop close relationships with their patients and subsequently experience significant grief after patient death. Consequences of this experience for employment outcomes have received little attention. To investigate staff, institutional, patient, and grief factors as predictors of burnout dimensions among direct care workers who had experienced recent patient death; determine which specific aspects of these factors are of particular importance; and establish grief as an independent predictor of burnout dimensions. Participants were 140 certified nursing assistants and 80 homecare workers who recently experienced patient death. Data collection involved comprehensive semistructured in-person interviews. Standardized assessments and structured questions addressed staff, patient, and institutional characteristics, grief symptoms and grief avoidance, as well as burnout dimensions (depersonalization, emotional exhaustion, and personal accomplishment). Hierarchical regressions revealed that grief factors accounted for unique variance in depersonalization, over and above staff, patient, and institutional factors. Supervisor support and caregiving benefits were consistently associated with higher levels on burnout dimensions. In contrast, coworker support was associated with a higher likelihood of depersonalization and emotional exhaustion. Findings suggest that grief over patient death plays an overlooked role in direct care worker burnout. High supervisor support and caregiving benefits may have protective effects with respect to burnout, whereas high coworker support may constitute a reflection of burnout. Copyright © 2017 American Academy of Hospice and Palliative Medicine. Published by Elsevier Inc. All rights reserved.

Long-term degradation of enamel and dentin bonds: 6-year results in vitro vs. in vivo

NARCIS (Netherlands)

Garcia-Godoy, F.; Krämer, N.; Feilzer, A.J.; Frankenberger, R.

2010-01-01

Objectives To evaluate marginal integrity of direct resin composite restorations before and after thermo-mechanical loading in vitro, and before and after 6 years of clinical service in a prospective clinical trial. Methods For the in vitro part, MO cavities with the proximal box beneath the

Direct assessment of lipoprotein outflow from in vivo-labeled arterial tissue as determined in an in vitro perfusion system.

Science.gov (United States)

Björnheden, T; Bondjers, G; Wiklund, O

1998-12-01

The rate of cholesterol deposition during the atherosclerotic process is determined by the balance between the inflow and outflow of plasma lipoproteins in the arterial wall. Whereas the rate of inflow may be measured directly, the rate of outflow has most often been calculated indirectly from lipoprotein uptake by using the 2-compartment model. One objection against such calculations is that lipoprotein binding is not being considered. In the present study 2 different protocols were used to obtain a direct measure of the outflow of lipoproteins from atherosclerotic rabbit aortas. Thus, 3 rabbits with experimental atherosclerosis were given 125I-LDL intravenously and 3 were given [14C]cholesterol perorally. Twenty-four hours later the aortas were removed and the outflow of label was monitored during in vitro perfusion. Despite the different protocols, our results were consistent and indicated that fractional loss relative to whole tissue was approximately 0.01 pool/h, which is 1 order of magnitude lower than current estimates based on the 2-compartment model (0.04 to 0.4 pool/h). Furthermore, whereas as much as 2/3 to 3/4 of the tracer that had entered the arterial wall was effectively trapped, the remainder equilibrated at a faster rate (0.06 pool/h). In conclusion, it seems that tissue binding constitutes a prominent and possibly underrated mechanism of lipoprotein deposition, at least in the atherosclerotic rabbit aorta. Furthermore, this means that current estimates of lipoprotein exchange parameters based on the 2-compartment model (eg, fractional loss) may rest on invalid assumptions and should be regarded with caution.

An adjustable gas-mixing device to increase feasibility of in vitro culture of Plasmodium falciparum parasites in the field.

Directory of Open Access Journals (Sweden)

Amy K Bei

Full Text Available A challenge to conducting high-impact and reproducible studies of the mechanisms of P. falciparum drug resistance, invasion, virulence, and immunity is the lack of robust and sustainable in vitro culture in the field. While the technology exists and is routinely utilized in developed countries, various factors-from cost, to supply, to quality-make it hard to implement in malaria endemic countries. Here, we design and rigorously evaluate an adjustable gas-mixing device for the in vitro culture of P. falciparum parasites in the field to circumvent this challenge. The device accurately replicates the gas concentrations needed to culture laboratory isolates, short-term adapted field isolates, cryopreserved previously non-adapted isolates, as well as to adapt ex vivo isolates to in vitro culture in the field. We also show an advantage over existing alternatives both in cost and in supply. Furthermore, the adjustable nature of the device makes it an ideal tool for many applications in which varied gas concentrations could be critical to culture success. This adjustable gas-mixing device will dramatically improve the feasibility of in vitro culture of Plasmodium falciparum parasites in malaria endemic countries given its numerous advantages.

Non-targeted and delayed effects of exposure to ionizing radiation: I. Radiation-induced genomic instability and bystander effects in vitro

Science.gov (United States)

Morgan, William F.

2003-01-01

A long-standing dogma in the radiation sciences is that energy from radiation must be deposited in the cell nucleus to elicit a biological effect. A number of non-targeted, delayed effects of ionizing radiation have been described that challenge this dogma and pose new challenges to evaluating potential hazards associated with radiation exposure. These effects include induced genomic instability and non-targeted bystander effects. The in vitro evidence for non-targeted effects in radiation biology will be reviewed, but the question as to how one extrapolates from these in vitro observations to the risk of radiation-induced adverse health effects such as cancer remains open.

Weak Static and Extremely Low Frequency Magnetic Fields Affect In Vitro Pollen Germination

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Lucietta Betti

2011-01-01

Full Text Available This study concerns the effects of a weak static magnetic field (MF at 10 μT oriented downward, combined with a 16-Hz sinusoidal MF (10 μT, on in vitro pollen germination of kiwifruit (Actinidia deliciosa. Extremely low frequency magnetic field (ELF-MF exposure was carried out by a signal generator unit connected to a copper wire solenoid, inside which samples where placed. Two different kinds of treatment were performed: direct and indirect. In the direct treatment, pollen samples were directly exposed during rehydration, germination, or both. In the indirect treatment, the pollen growth medium was prepared with water aliquots (at standard temperature of 20°C and pH = 6.74 that were exposed before use for 8 or 24 h. The main purpose of our research was to identify a biological marker (in vitro pollen germination in a stressing growth medium without Ca2+ susceptible to the effects of direct or indirect ELF-MF exposure. The working variable was the pollen germination rate, as detected blind after 3 h 30 min by an Axioplan microscope. A directionally consistent recovery of germination percentage was observed both for direct exposure (during germination and both rehydration and germination phases and water-mediated exposure (with water exposed for 24 h and immediately used. Our results suggest that the ELF-MF treatment might partially remove the inhibitory effect caused by the lack of Ca2+ in the culture medium, inducing a release of internal Ca2+ stored in the secretory vesicles of pollen plasma membrane. Although preliminary, findings seem to indicate the in vitro pollen performance as adequate to study the effects of ELF-MFs on living matter.

Surface disinfection challenges for Candida auris: an in-vitro study.

Science.gov (United States)

Kean, R; Sherry, L; Townsend, E; McKloud, E; Short, B; Akinbobola, A; Mackay, W G; Williams, C; Jones, B L; Ramage, G

2018-04-01

The emerging pathogenic multidrug-resistant yeast Candida auris is an important source of healthcare-associated infections and of growing global clinical concern. The ability of this organism to survive on surfaces and withstand environmental stressors creates a challenge for eradicating it from hospitals. A panel of C. auris clinical isolates was evaluated on different surface environments against the standard disinfectant sodium hypochlorite and high-level disinfectant peracetic acid. C. auris was shown to selectively tolerate clinically relevant concentrations of sodium hypochlorite and peracetic acid in a surface-dependent manner, which may explain its ability to successfully persist within the hospital environment. Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Problems in extrapolating genotoxicity data from cellular systems in vitro

Energy Technology Data Exchange (ETDEWEB)

Mohn, G.R.; van Zeeland, A.A.

1986-01-01

The experiments involved quantitative comparative mutagenesis determinations using ethylating agents differing in DNA reactivity, mammalian metabolism, and mutagenic potency. The results can be summarized as follows: (1) Under standardized in vitro treatment conditions and at identical DNA dose levels, the frequencies of induced mutations observed in the present bacterial systems are not representative and usually lower than those induced in cultured mammalian cells. (2) Under the same in vitro treatment conditions, the relative mutagenic potency order is identical in bacteria and the mammalian cells, namely, N-ethyl-N'-nitrosoquinioline(ENNG) > ethylnitrosourea (ENU) > diethylsulfate (DES) > ethylmethanesulfonate (EMS); diethylnitrosamine (DEN) being not mutagenic in the two organisms without addition of mammalian (rodent) liver preparations containing Cyt-P450-dependent mixed function oxidases. (3) The extremely high mutagenic activity of ENNG in bacteria and mammalian cells in vitro is very probably due to its intracellular, glutathione-mediated activation to highly reactive chemical species. (4) Both in bacteria and the mammalian cells the frequencies of induced mutations are directly proportional to the levels of O6-ethylguanine formed in DNA after exposure to ENNG, ENU, DES, and EMS, in contrast to results obtained when other DNA adducts are used as dose parameters. (5) The addition of mouse liver S-9 preparations in the in vitro assays allows to determine which chemicals are likely to be substantially activated (DEN) or deactivated (ENNG) in vivo; however, the mutagenic potency order remained the same under these conditions. (6) When the indicator bacteria are exposed directly to the in vivo metabolism of living animals, the relative order of mutagenic potency on the basis of exposure levels becomes drastically different from that observed in vitro, using either E. coli or V79 mammalian cells.

Identification of a novel phosphorylation site in c-jun directly targeted in vitro by protein kinase D

International Nuclear Information System (INIS)

Waldron, Richard T.; Whitelegge, Julian P.; Faull, Kym F.; Rozengurt, Enrique

2007-01-01

Protein kinase D (PKD) phosphorylates the c-jun amino-terminal in vitro at site(s) distinct from JNK [C. Hurd, R.T. Waldron, E. Rozengurt, Protein kinase D complexes with c-jun N-terminal kinase via activation loop phosphorylation and phosphorylates the c-jun N-terminus, Oncogene 21 (2002) 2154-2160], but the sites have not been identified. Here, metabolic 32 P-labeling of c-jun protein in COS-7 cells indicated that PKD phosphorylates c-jun in vivo at a site(s) between aa 43-93, a region containing important functional elements. On this basis, the PKD-mediated phosphorylation site(s) was further characterized in vitro using GST-c-jun fusion proteins. PKD did not incorporate phosphate into Ser63 and Ser73, the JNK sites in GST-c-jun(1-89). Rather, PKD and JNK could sequentially phosphorylate distinct site(s) simultaneously. By mass spectrometry of tryptic phosphopeptides, Ser58 interposed between the JNK-binding portion of the delta domain and the adjacent TAD1 was identified as a prominent site phosphorylated in vitro by PKD. These data were further supported by kinase reactions using truncations or point-mutations of GST-c-jun. Together, these data suggest that PKD-mediated phosphorylation modulates c-jun at the level of its N-terminal functional domains

Torque resistance of impression copings after direct implant impression: An in vitro evaluation of impression materials with and without adhesive.

Science.gov (United States)

Auroy, Pascal; Nicolas, Emanuel; Bedouin, Yvan

2017-01-01

No data are available on the ability of an impression coping to resist the manual placement of an abutment replica (implant analog) during prosthodontic laboratory procedures after a direct (pick-up) impression. The purpose of this in vitro study was to evaluate the torque resistance of impression copings after a direct impression, that is, the amount of rotational torque sufficient to induce irreversible displacement of impression copings in the impression material bulk once the impression has been made. A reference model with 5 abutment replicas was constructed. Five impression copings were screwed onto the abutment replicas, and standardized impressions were made. A controlled twisting force was applied to each impression coping. A torque tester recorded the torque variation. Three elastomeric impression materials were tested. ANOVA and the Tukey test (α=.05) were performed using an average of 30 measurements per impression material, with and without adhesive. ANOVA and the Tukey test results showed that the adhesive, cohesive, and mechanical bonds between the impression coping and the impression material depended greatly on the type of material and that the average rupture threshold of these bonds was statistically significantly different in pairwise comparisons (Ptorque. The polyether impression material is the direct impression material that showed the highest breakdown threshold for adhesive bonding when used without an adhesive. The use of an adhesive on impression copings leads to irreversible deformation of the interface at torque stresses well below the adhesive bond threshold of the same materials used without an adhesive. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

4R-Cembranoid Improves Outcomes after 6-Hydroxydopamine Challenge in Both In vitro and In vivo Models of Parkinson's Disease

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Jing Hu

2017-05-01

Full Text Available (1S, 2E, 4R, 6R,-7E, 11E-2, 7, 11-cembratriene-4, 6-diol (4R is one of the cembranoids found in tobacco leaves. Previous studies have found that 4R protected acute rat hippocampal slices against neurotoxicity induced by N-methyl-D-aspartate (NMDA and against the toxic organophosphorus compounds paraoxon and diisopropylfluorophosphate (DFP. Furthermore, in vivo, 4R reduced the infarct size in a rodent ischemic stroke model and neurodegeneration caused by DFP. The present study expanded our previous study by focusing on the effect of 4R in Parkinson's disease (PD and elucidating its underlying mechanisms using 6-hydroxydopamine (6-OHDA-induced injury models. We found that 4R exhibited significant neuroprotective activity in the rat unilateral 6-OHDA-induced PD model in vivo. The therapeutic effect was evident both at morphological and behavioral levels. 4R (6 and 12 mg/kg treatments significantly improved outcomes of 6-OHDA-induced PD in vivo as indicated by reducing forelimb asymmetry scores and corner test scores 4 weeks after injection of 6-OHDA (p < 0.05. The therapeutic effect of 4R was also reflected by decreased depletion of tyrosine hydroxylase (TH in the striatum and substantia nigra (SN on the side injected with 6-OHDA. TH expression was 70.3 and 62.8% of the contralateral side in striatum and SN, respectively, after 6 mg/kg 4R treatment; furthermore, it was 80.1 and 79.3% after treatment with 12 mg/kg of 4R. In the control group, it was 51.9 and 23.6% of the contralateral striatum and SN (p < 0.05. Moreover, 4R also protected differentiated neuro-2a cells from 6-OHDA-induced cytotoxicity in vitro. The activation of p-AKT and HAX-1, and inhibition of caspase-3 and endothelial inflammation, were involved in 4R-mediated protection against 6-OHDA-induced injury. In conclusion, the present study indicates that 4R shows a therapeutic effect in the rat 6-OHDA-induced PD model in vivo and in 6-OHDA-challenged neuro-2a cells in vitro.

Workshop proceedings: challenges and opportunities in evaluating protein allergenicity across biotechnology industries.

Science.gov (United States)

Stagg, Nicola J; Ghantous, Hanan N; Ladics, Gregory S; House, Robert V; Gendel, Steven M; Hastings, Kenneth L

2013-01-01

A workshop entitled "Challenges and Opportunities in Evaluating Protein Allergenicity across Biotechnology Industries" was held at the 51st Annual Meeting of the Society of Toxicology (SOT) in San Francisco, California. The workshop was sponsored by the Biotechnology Specialty Section of SOT and was designed to present the science-based approaches used in biotechnology industries to evaluate and regulate protein allergenicity. A panel of experts from industry and government highlighted the allergenicity testing requirements and research in the agricultural, pharmaceutical/biopharma, and vaccine biotechnology industries and addressed challenges and opportunities for advancing the science of protein allergenicity. The main learning from the workshop was that immunoglobulin E-mediated allergenicity of biotechnology-derived products is difficult to assess without human data. The approaches currently being used to evaluate potential for allergenicity across biotechnology industries are very different and range from bioinformatics, in vitro serology, in vivo animal testing, in vitro and in vivo functional assays, and "biosimilar" assessments (ie, biotherapeutic equivalents to innovator products). The challenge remains with regard to the different or lack of regulatory requirements for allergenicity testing across industries, but the novel approaches being used with bioinformatics and biosimilars may lead to opportunities in the future to collaborate across biotechnology industries.

Labeling and stability of radiolabeled antibody fragments by a direct 99mTc-labeling method

International Nuclear Information System (INIS)

Pak, K.Y.; Nedelman, M.A.; Tam, S.H.; Wilson, E.; Daddona, P.E.

1992-01-01

The in vitro labeling and stability of 99m Tc-labeled antibody Fab' fragments prepared by a direct labeling technique were evaluated. Eight antibody fragments derived from murine IgG1 (N = 5), IgG2a (N = 2) and IgG3 (N = 1) isotypes were labeled with a preformed 99m Tc-D-glucarate complex. No loss of radioactivity incorporation was observed for all the 99m Tc-labeled antibody fragments after 24 h incubation at 37 o C. 99m Tc-labeled antibody fragments (IgG1, N = 2; IgG2a, n = 2; IgG3, N = 1) were stable upon challenge with DTPA, EDTA or acidic pH. Using the affinity chromatography technique, two of the 99m Tc-labeled antibody fragments displayed no loss of immunoreactivity after prolonged incubation in phosphate buffer up to 24 h at 37 o C. Bonding between 99m Tc and antibody fragments was elucidated by challenging with a diamide ditholate (N 2 S 2 ) compound. The Fab' with IgG2a isotype displayed tighter binding to 99m Tc in comparison to Fab' from IgG1 and IgG3 isotype in N 2 S 2 challenge and incubation with human plasma. The in vivo biodistribution of five 99m Tc-labeled fragments were evaluated in normal mice. (Author)

Immunogenicity of ascites tumor cells following in vitro hyperthermia

International Nuclear Information System (INIS)

Dickson, J.A.; Jasiewicz, M.L.; Simpson, A.C.

1982-01-01

The concept that host immunization may be achieved by heat-induced antigenic modifications of cancer cells and/or the release of immunogenic products by dead or dying tumor cells following in vitro heating was examined. Ehrlich ascites cells were used, inasmuch as it was claimed that in vitro hyperthermia increased the immunogenicity of these cells. Tumor cell populations of different viability were obtained by heating Ehrlich cells at 42.5 degrees, 45 degrees, or 60 degrees C. Viable and nonviable cells were separated by Ficoll-Hypaque density centrifugation; viable nonreplicating cells were obtained by treatment with mitomycin C. Cell populations of different viability after heating were left to die slowly over 3 days at 37 degrees C. Swiss TO mice were then given injections of the treated cells and/or medium. No survival benefit occurred in mice inoculated with any of these different components and then challenged with viable tumor cells. Injection of irradiated cells, however, did produce host immunity. Similarly, D23 rat hepatoma ascites cells produced host immunity after 15,000 rad but not after heating. The claim that in vitro hyperthermia increases the immunogenicity of tumor cells was not confirmed

Current challenges and future directions for bacterial self-healing concrete.

Science.gov (United States)

Lee, Yun Suk; Park, Woojun

2018-04-01

Microbially induced calcium carbonate precipitation (MICP) has been widely explored and applied in the field of environmental engineering over the last decade. Calcium carbonate is naturally precipitated as a byproduct of various microbial metabolic activities. This biological process was brought into practical use to restore construction materials, strengthen and remediate soil, and sequester carbon. MICP has also been extensively examined for applications in self-healing concrete. Biogenic crack repair helps mitigate the high maintenance costs of concrete in an eco-friendly manner. In this process, calcium carbonate precipitation (CCP)-capable bacteria and nutrients are embedded inside the concrete. These bacteria are expected to increase the durability of the concrete by precipitating calcium carbonate in situ to heal cracks that develop in the concrete. However, several challenges exist with respect to embedding such bacteria; harsh conditions in concrete matrices are unsuitable for bacterial life, including high alkalinity (pH up to 13), high temperatures during manufacturing processes, and limited oxygen supply. Additionally, many biological factors, including the optimum conditions for MICP, the molecular mechanisms involved in MICP, the specific microorganisms suitable for application in concrete, the survival characteristics of the microorganisms embedded in concrete, and the amount of MICP in concrete, remain unclear. In this paper, metabolic pathways that result in conditions favorable for calcium carbonate precipitation, current and potential applications in concrete, and the remaining biological challenges are reviewed.

Challenges in Translational Development of Pharmaceutical Cocrystals.

Science.gov (United States)

Kale, Dnyaneshwar P; Zode, Sandeep S; Bansal, Arvind K

2017-02-01

The last 2 decades have witnessed increased research in the area of cocrystals resulting in deeper scientific understanding, increase in intellectual property landscape, and evolution in the regulatory environment. Pharmaceutical cocrystals have received significant attention as a new solid form on account of their ability to modulate poor physicochemical properties of drug molecules. However, pharmaceutical development of cocrystals could be challenging, thus limiting their translation into viable drug products. In the present commentary, the role of cocrystals in the modulation of material properties and challenges involved in the pharmaceutical development of cocrystals have been discussed. The major hurdles encountered in the development of cocrystals such as safety of coformers, unpredictable performance during dissolution and solubility in different media, difficulties in establishing in vitro-in vivo correlation, and polymorphism have been extensively discussed. The influence of selecting appropriate formulation and process design on these challenges has been discussed. Finally, a brief outline of cocrystals that are undergoing clinical development has also been presented. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Twenty-first century metadata operations challenges, opportunities, directions

CERN Document Server

Lee Eden, Bradford

2014-01-01

It has long been apparent to academic library administrators that the current technical services operations within libraries need to be redirected and refocused in terms of both format priorities and human resources. A number of developments and directions have made this reorganization imperative, many of which have been accelerated by the current economic crisis. All of the chapters detail some aspect of technical services reorganization due to downsizing and/or reallocation of human resources, retooling professional and support staff in higher level duties and/or non-MARC metadata, ""value-a

Challenges to implementation of advance directives of will in hospital practice.

Science.gov (United States)

Cogo, Silvana Bastos; Lunardi, Valéria Lerch; Quintana, Alberto Manuel; Girardon-Perlini, Nara Marilene Oliveira; Silveira, Rosemary Silva da

2016-01-01

to understand the difficulties and limitations in the implementation of advance directives of will in the hospital context. qualitative, exploratory and descriptive study conducted by means of semi-structured interviews with nurses, resident physicians and family caregivers. The data were analyzed by using discursive textual analysis based on the framework of bioethics principles. the following categories emerged: Terminality as an expression of loss and cure as an option for care; concerns about legal implications; advance directives of will demand patient autonomy and proper communication. limitations and difficulties in practice of advance directives of will from the perspective of the participants show, apart from countless conflicts and dilemmas regarding end-of life matters, that impending death experiences obstruct patients' wishes.

AR-13, a Celecoxib Derivative, Directly Kills Francisella In Vitro and Aids Clearance and Mouse Survival In Vivo

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Ky V. Hoang

2017-09-01

Full Text Available Francisella tularensis (F. tularensis is the causative agent of tularemia and is classified as a Tier 1 select agent. No licensed vaccine is currently available in the United States and treatment of tularemia is confined to few antibiotics. In this study, we demonstrate that AR-13, a derivative of the cyclooxygenase-2 inhibitor celecoxib, exhibits direct in vitro bactericidal killing activity against Francisella including a type A strain of F. tularensis (SchuS4 and the live vaccine strain (LVS, as well as toward the intracellular proliferation of LVS in macrophages, without causing significant host cell toxicity. Identification of an AR-13-resistant isolate indicates that this compound has an intracellular target(s and that efflux pumps can mediate AR-13 resistance. In the mouse model of tularemia, AR-13 treatment protected 50% of the mice from lethal LVS infection and prolonged survival time from a lethal dose of F. tularensis SchuS4. Combination of AR-13 with a sub-optimal dose of gentamicin protected 60% of F. tularensis SchuS4-infected mice from death. Taken together, these data support the translational potential of AR-13 as a lead compound for the further development of new anti-Francisella agents.

Game Theory-Based Cooperation for Underwater Acoustic Sensor Networks: Taxonomy, Review, Research Challenges and Directions.

Science.gov (United States)

Muhammed, Dalhatu; Anisi, Mohammad Hossein; Zareei, Mahdi; Vargas-Rosales, Cesar; Khan, Anwar

2018-02-01

Exploring and monitoring the underwater world using underwater sensors is drawing a lot of attention these days. In this field cooperation between acoustic sensor nodes has been a critical problem due to the challenging features such as acoustic channel failure (sound signal), long propagation delay of acoustic signal, limited bandwidth and loss of connectivity. There are several proposed methods to improve cooperation between the nodes by incorporating information/game theory in the node's cooperation. However, there is a need to classify the existing works and demonstrate their performance in addressing the cooperation issue. In this paper, we have conducted a review to investigate various factors affecting cooperation in underwater acoustic sensor networks. We study various cooperation techniques used for underwater acoustic sensor networks from different perspectives, with a concentration on communication reliability, energy consumption, and security and present a taxonomy for underwater cooperation. Moreover, we further review how the game theory can be applied to make the nodes cooperate with each other. We further analyze different cooperative game methods, where their performance on different metrics is compared. Finally, open issues and future research direction in underwater acoustic sensor networks are highlighted.

Game Theory-Based Cooperation for Underwater Acoustic Sensor Networks: Taxonomy, Review, Research Challenges and Directions

Directory of Open Access Journals (Sweden)

Dalhatu Muhammed

2018-02-01

Full Text Available Exploring and monitoring the underwater world using underwater sensors is drawing a lot of attention these days. In this field cooperation between acoustic sensor nodes has been a critical problem due to the challenging features such as acoustic channel failure (sound signal, long propagation delay of acoustic signal, limited bandwidth and loss of connectivity. There are several proposed methods to improve cooperation between the nodes by incorporating information/game theory in the node’s cooperation. However, there is a need to classify the existing works and demonstrate their performance in addressing the cooperation issue. In this paper, we have conducted a review to investigate various factors affecting cooperation in underwater acoustic sensor networks. We study various cooperation techniques used for underwater acoustic sensor networks from different perspectives, with a concentration on communication reliability, energy consumption, and security and present a taxonomy for underwater cooperation. Moreover, we further review how the game theory can be applied to make the nodes cooperate with each other. We further analyze different cooperative game methods, where their performance on different metrics is compared. Finally, open issues and future research direction in underwater acoustic sensor networks are highlighted.

Game Theory-Based Cooperation for Underwater Acoustic Sensor Networks: Taxonomy, Review, Research Challenges and Directions

Science.gov (United States)

Muhammed, Dalhatu; Anisi, Mohammad Hossein; Vargas-Rosales, Cesar; Khan, Anwar

2018-01-01

Exploring and monitoring the underwater world using underwater sensors is drawing a lot of attention these days. In this field cooperation between acoustic sensor nodes has been a critical problem due to the challenging features such as acoustic channel failure (sound signal), long propagation delay of acoustic signal, limited bandwidth and loss of connectivity. There are several proposed methods to improve cooperation between the nodes by incorporating information/game theory in the node’s cooperation. However, there is a need to classify the existing works and demonstrate their performance in addressing the cooperation issue. In this paper, we have conducted a review to investigate various factors affecting cooperation in underwater acoustic sensor networks. We study various cooperation techniques used for underwater acoustic sensor networks from different perspectives, with a concentration on communication reliability, energy consumption, and security and present a taxonomy for underwater cooperation. Moreover, we further review how the game theory can be applied to make the nodes cooperate with each other. We further analyze different cooperative game methods, where their performance on different metrics is compared. Finally, open issues and future research direction in underwater acoustic sensor networks are highlighted. PMID:29389874

Gastric ulcer localization by direct in vivo labeling of sucralfate: work in progress

Energy Technology Data Exchange (ETDEWEB)

Pera, A.; Seevers, R.H.; Meyer, K.; Hall, C.; Bekerman, C.; Anderson, T.M.; Katzen, H.; Laakso, L.; Pinsky, S.M.

1985-09-01

The authors developed and evaluated a new procedure for imaging gastric ulcer disease with technetium 99m - labeled sucralfate. The new method employs direct in vivo labeling of sucralfate instead of in vitro labeling using human serum albumin, as previously reported in the literature. In 26 studies using humans with sucralfate labeled directly in vivo, 15 gave true-negative results. Of 14 studies using humans with in vitro labeled sucralfate, three gave true-negative results, three gave true-positive results, and the results of eight were either false-negative or could not be interpreted because of high levels of activity remaining in the stomach. They suggest that the direct in vivo labeling method significantly improves the sucralfate gastric ulcer imaging technique.

Gastric ulcer localization by direct in vivo labeling of sucralfate: work in progress

International Nuclear Information System (INIS)

Pera, A.; Seevers, R.H.; Meyer, K.; Hall, C.; Bekerman, C.; Anderson, T.M.; Katzen, H.; Laakso, L.; Pinsky, S.M.

1985-01-01

The authors developed and evaluated a new procedure for imaging gastric ulcer disease with technetium 99m - labeled sucralfate. The new method employs direct in vivo labeling of sucralfate instead of in vitro labeling using human serum albumin, as previously reported in the literature. In 26 studies using humans with sucralfate labeled directly in vivo, 15 gave true-negative results. Of 14 studies using humans with in vitro labeled sucralfate, three gave true-negative results, three gave true-positive results, and the results of eight were either false-negative or could not be interpreted because of high levels of activity remaining in the stomach. They suggest that the direct in vivo labeling method significantly improves the sucralfate gastric ulcer imaging technique

Direct organogenesis and leaf-anatomy modifications in vitro of neoregelia concentrica (vellozo) l.b. smith (bromeliaceae)

International Nuclear Information System (INIS)

Martins, J.P.R.; Castro, E.M.D.; Jani, T.F.; Pasqual, M.; Schmildt, E.R.

2014-01-01

Tissue culture can contribute in the multiplication of several species with commercial interest, like the bromeliads. It was aimed to evaluate cytokinins and its concentrations in the multiplication and leaf structure of Neoregelia concentrica (Vellozo) L.B. Smith. Previously In vitro-established N. concentrica plants were inoculated in MS medium supplemented with 6-benzylaminopurine (BAP) or Kinetin (KIN) with concentrations 0.0, 5.0, 10.0 and 15.0 micro M. For the anatomic analyses tree plants of each treatment were randomly sampled at 60-day growth. Significant differences were verified in the evaluated characteristics due to the treatments. The raise in cytokinin concentrations induced a higher percentage and average number of explants with shoots. BAP provided higher averages when compared to KIN. The cytokinin use modified the epidermal structure and induced a larger thickening of the water-storage and chlorophyll parenchymas. The use of 15.0 micro M BAP was efficient in the In vitro multiplication and in the leaf tissue development of N. concentrica. (author)

Investigating biological traces of traumatic stress in changing societies: challenges and directions from the ESTSS Task Force on Neurobiology

Directory of Open Access Journals (Sweden)

Kathleen Thomaes

2016-03-01

Full Text Available Traumatic stress can have severe consequences for both mental and physical health. Furthermore, both psychological and biological traces of trauma increase as a function of accumulating traumatic experiences. Neurobiological research may aid in limiting the impact of traumatic stress, by leading to advances in preventive and treatment interventions. To promote the possibility for clinical implementation of novel research findings, this brief review describes timely conceptual and methodological challenges and directions in neurobiological trauma research on behalf of the Task Force “Neurobiology of Traumatic Stress” of the European Society for Traumatic Stress Studies (ESTSS. The most important conceptual challenges are the heterogeneity of disorders and existence of subtypes across diagnostic categories: differential latent profiles and trajectories regarding symptom expression and neural correlates are being unraveled; however, similar latent classes’ approaches for treatment response and neurobiological data remain scarce thus far. The key to improving the efficacy of currently available preventive interventions and treatments for trauma-related disorders lies in a better understanding and characterization of individual differences in response to trauma and interventions. This could lead to personalized treatment strategies for trauma-related disorders, based on objective information indicating whether individuals are expected to benefit from them. The most important methodological challenge identified here is the need for large consortia and meta-analyses or, rather, mega-analyses on existent data as a first step. In addition, large multicenter studies, combining novel methods for repeated sampling with more advanced statistical modeling techniques, such as machine learning, should aim to translate identified disease mechanisms into molecular blood-based biomarker combinations to predict disorder vulnerability and treatment responses.

Preparation of 2 nm gold nanoparticles for in vitro and in vivo applications

OpenAIRE

Moyano, Daniel F.; Duncan, Bradley; Rotello, Vincent M.

2013-01-01

Gold nanoparticles have been a versatile tool in recent years for the exploration of biological systems. However, challenges with purification and adequate surface coverage limit the biocompatibility of gold nanoparticles. Here, we describe a detailed procedure for the synthesis, purification, and functionalization of biologically compatible gold nanoparticles for in vitro and in vivo studies.

Statistical methods and challenges in connectome genetics

KAUST Repository

Pluta, Dustin

2018-03-12

The study of genetic influences on brain connectivity, known as connectome genetics, is an exciting new direction of research in imaging genetics. We here review recent results and current statistical methods in this area, and discuss some of the persistent challenges and possible directions for future work.

Blood vessel endothelium-directed tumor cell streaming in breast tumors requires the HGF/C-Met signaling pathway.

Science.gov (United States)

Leung, E; Xue, A; Wang, Y; Rougerie, P; Sharma, V P; Eddy, R; Cox, D; Condeelis, J

2017-05-11

During metastasis to distant sites, tumor cells migrate to blood vessels. In vivo, breast tumor cells utilize a specialized mode of migration known as streaming, where a linear assembly of tumor cells migrate directionally towards blood vessels on fibronectin-collagen I-containing extracellular matrix (ECM) fibers in response to chemotactic signals. We have successfully reconstructed tumor cell streaming in vitro by co-plating tumors cells, macrophages and endothelial cells on 2.5 μm thick ECM-coated micro-patterned substrates. We found that tumor cells and macrophages, when plated together on the micro-patterned substrates, do not demonstrate sustained directional migration in only one direction (sustained directionality) but show random bi-directional walking. Sustained directionality of tumor cells as seen in vivo was established in vitro when beads coated with human umbilical vein endothelial cells were placed at one end of the micro-patterned 'ECM fibers' within the assay. We demonstrated that these endothelial cells supply the hepatocyte growth factor (HGF) required for the chemotactic gradient responsible for sustained directionality. Using this in vitro reconstituted streaming system, we found that directional streaming is dependent on, and most effectively blocked, by inhibiting the HGF/C-Met signaling pathway between endothelial cells and tumor cells. Key observations made with the in vitro reconstituted system implicating C-Met signaling were confirmed in vivo in mammary tumors using the in vivo invasion assay and intravital multiphoton imaging of tumor cell streaming. These results establish HGF/C-Met as a central organizing signal in blood vessel-directed tumor cell migration in vivo and highlight a promising role for C-Met inhibitors in blocking tumor cell streaming and metastasis in vivo, and for use in human trials.

In vitro corrosion of pure magnesium and AZ91 alloy?the influence of thin electrolyte layer thickness

OpenAIRE

Zeng, Rong-Chang; Qi, Wei-Chen; Zhang, Fen; Li, Shuo-Qi

2016-01-01

In vivo degradation predication faces a huge challenge via in vitro corrosion test due to the difficulty for mimicking the complicated microenvironment with various influencing factors. A thin electrolyte layer (TEL) cell for in vitro corrosion of pure magnesium and AZ91 alloy was presented to stimulate the in vivo corrosion in the micro-environment built by the interface of the implant and its neighboring tissue. The results demonstrated that the in vivo corrosion of pure Mg and the AZ91 all...

Subunit Organisation of In Vitro Reconstituted HOPS and CORVET Multisubunit Membrane Tethering Complexes

Science.gov (United States)

Guo, Zhong; Johnston, Wayne; Kovtun, Oleksiy; Mureev, Sergey; Bröcker, Cornelia; Ungermann, Christian; Alexandrov, Kirill

2013-01-01

Biochemical and structural analysis of macromolecular protein assemblies remains challenging due to technical difficulties in recombinant expression, engineering and reconstitution of multisubunit complexes. Here we use a recently developed cell-free protein expression system based on the protozoan Leishmania tarentolae to produce in vitro all six subunits of the 600 kDa HOPS and CORVET membrane tethering complexes. We demonstrate that both subcomplexes and the entire HOPS complex can be reconstituted in vitro resulting in a comprehensive subunit interaction map. To our knowledge this is the largest eukaryotic protein complex in vitro reconstituted to date. Using the truncation and interaction analysis, we demonstrate that the complex is assembled through short hydrophobic sequences located in the C-terminus of the individual Vps subunits. Based on this data we propose a model of the HOPS and CORVET complex assembly that reconciles the available biochemical and structural data. PMID:24312556

Direct measurement of the poliovirus RNA polymerase error frequency in vitro

International Nuclear Information System (INIS)

Ward, C.D.; Stokes, M.A.M.; Flanegan, J.B.

1988-01-01

The fidelity of RNA replication by the poliovirus-RNA-dependent RNA polymerase was examined by copying homopolymeric RNA templates in vitro. The poliovirus RNA polymerase was extensively purified and used to copy poly(A), poly(C), or poly(I) templates with equimolar concentrations of noncomplementary and complementary ribonucleotides. The error frequency was expressed as the amount of a noncomplementary nucleotide incorporated divided by the total amount of complementary and noncomplementary nucleotide incorporated. The polymerase error frequencies were very high, depending on the specific reaction conditions. The activity of the polymerase on poly(U) and poly(G) was too low to measure error frequencies on these templates. A fivefold increase in the error frequency was observed when the reaction conditions were changed from 3.0 mM Mg 2+ (pH 7.0) to 7.0 mM Mg 2+ (pH 8.0). This increase in the error frequency correlates with an eightfold increase in the elongation rate that was observed under the same conditions in a previous study

Creating Patient-Specific Neural Cells for the In Vitro Study of Brain Disorders

Directory of Open Access Journals (Sweden)

Kristen J. Brennand

2015-12-01

Full Text Available As a group, we met to discuss the current challenges for creating meaningful patient-specific in vitro models to study brain disorders. Although the convergence of findings between laboratories and patient cohorts provided us confidence and optimism that hiPSC-based platforms will inform future drug discovery efforts, a number of critical technical challenges remain. This opinion piece outlines our collective views on the current state of hiPSC-based disease modeling and discusses what we see to be the critical objectives that must be addressed collectively as a field.

State of the art and challenges in development of electrical contact materials in the light of the RoHS directive

Directory of Open Access Journals (Sweden)

Ćosović V.

2012-01-01

Full Text Available The article surveys current state of the art and challenges in the development of the electrical contact materials in the light of the EU Directive on Restriction of Hazardous Substances (RoHS. The focus was placed on widely used silver-cadmium alloys. According to this directive, as of July 1, 2006, use of six hazardous materials, including cadmium, are restricted for applicable electrical and electronic products intended for the EU market. In contrast, traditionally preferred material for production of electrical contacts is Ag-CdO, due to its outstanding functional properties. These conflicting interests result in present state where RoHS directive has not yet been implemented in its original form and has undergone numerous amendments and exceptions regarding the use of cadmium. Main reason for this seems to be the unrealistic time frame imposed by legislation. Although, significant effort has been put into research and development of alternative materials, there are still cases where adequate replacement materials had not been found. Therefore, importance of synchronicity between legislation and technological progress i.e. communication between legislative administration and industry and academia is brought to light as well as some common issues that may arise with an introduction of new replacement materials or product modification. [Projekat Ministarstva nauke Republike SRbije, br. OI 172037 and TR 34023

A Unifying Approach to Goal-Directed Evaluation

DEFF Research Database (Denmark)

Danvy, Olivier; Rhiger, Morten; Grobauer, Bernd

2001-01-01

Goal-directed evaluation, as embodied in Icon and Snobol, is built on the notions of backtracking and of generating successive results, and therefore it has always been something of a challenge to specify and implement. In this article, we address this challenge using computational monads and par...

Factors affecting survival rates of in vitro produced bovine embryos after vitrification and direct in-straw rehydration

DEFF Research Database (Denmark)

Vajta, G.; Holm, P.; Greve, T.

1996-01-01

%) and no hatching of these embryos was observed. In the second experiment, Day 7 expanded blastocysts were vitrified using PBS, PBS + albumin, TCM199 and TCM 199 + calf serum as holding media. No differences in re-expansion and hatching rates were seen. However, when incubation with the concentrated cryoprotectant......The aim of this work was to investigate the possibilities of simplification, and to outline the limits of application, of a vitrification method for cow embryos. Morulae and blastocysts were produced by in vitro fertilization of slaughterhouse-derived, in vitro matured oocytes with frozen...... and developmental stage (Day 5 compacted morulae, Day 6 early blastocysts, Days 6 and 7 blastocysts, Day 7 expanded blastocysts and Day 8 hatched blastocysts) as well as Days 7 and 5 blastocysts previously subjected to partial zona dissection were vitrified. After thawing, the re-expansion rates of blastocysts...

Shark Interactions With Directed and Incidental Fisheries in the Northeast Pacific Ocean: Historic and Current Encounters, and Challenges for Shark Conservation.

Science.gov (United States)

King, Jackie; McFarlane, Gordon A; Gertseva, Vladlena; Gasper, Jason; Matson, Sean; Tribuzio, Cindy A

For over 100 years, sharks have been encountered, as either directed catch or incidental catch, in commercial fisheries throughout the Northeast Pacific Ocean. A long-standing directed fishery for North Pacific Spiny Dogfish (Squalus suckleyi) has occurred and dominated shark landings and discards. Other fisheries, mainly for shark livers, have historically targeted species including Bluntnose Sixgill Shark (Hexanchus griseus) and Tope Shark (Galeorhinus galeus). While incidental catches of numerous species have occurred historically, only recently have these encounters been reliably enumerated in commercial and recreational fisheries. In this chapter we present shark catch statistics (directed and incidental) for commercial and recreational fisheries from Canadian waters (off British Columbia), southern US waters (off California, Oregon, and Washington), and northern US waters (off Alaska). In total, 17 species of sharks have collectively been encountered in these waters. Fishery encounters present conservation challenges for shark management, namely, the need for accurate catch statistics, stock delineation, life history parameter estimates, and improved assessments methods for population status and trends. Improvements in management and conservation of shark populations will only come with the further development of sound science-based fishery management practices for both targeted and incidental shark fisheries. © 2017 Elsevier Ltd All rights reserved.

The Rise of Direct Democracy in Croatia: Balancing or Challenging Parliamentary Representation?

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Butković Hrvoje

2017-03-01

Full Text Available In 2010 the Croatian Constitution was changed to lower the requirements for the implementation of direct democracy at the national level, in order to save the referendum on Croatia’s EU membership from possible failure. Since then, Croatia has witnessed a sharp increase in people’s initiatives that have managed to block a number of the government’s reform proposals. Therefore, the newly discovered appeal of direct democracy in Croatia has created a new environment for the operation of its representative democracy. Starting from theoretical notions, this paper analyses the practice of direct democracy in selected transitional countries, which could be instructive for Croatia. In its central part, the paper explores the obstacles that stand in the way of the efficient implementation of direct democracy in Croatia.

[Intragastric provocation and antigen-induced in vitro histamine liberation by the food additive E 102].

Science.gov (United States)

Schaubschläger, W; Ruschmeyer, J; Zabel, P; Schlaak, M

1988-06-01

Adverse reactions to tartrazine have been known since 1958. The mechanism of this reaction, a not IgE-mediated, anaphylactoid reaction, is not fully understood. The demonstration of this adverse reaction by provocative challenge feeding may be problematic by a score of subjective symptoms because of the placebo effect. This report deals with the intragastral provocation under endoscopic control with tartrazine and tartrazine-induced histamine release in vitro from gastric mucosa and from blood. Two patients with anamnestically suspected adverse reactions to tartrazine were studied. Correspondence of in vivo and in vitro testing with tartrazine could be demonstrated.

Analysis of Chemical Bioactivity through In Vitro Profiling ...

Science.gov (United States)

Safety assessment of drugs and environmental chemicals relies extensively on animal testing. However, the quantity of chemicals needing assessment and challenges of species extrapolation drive the development of alternative approaches. The EPA’s ToxCast and the multiagency Tox21 programs address this through use of an extensive in vitro screening program to generate data on a large library of important environmental chemicals. These in vitro assays encompass both cell-free, biochemical assays targeting proteins that may be potential molecular initiating events and cellular assays that provide coverage of critical signaling pathways and toxicity phenotypes. Effects on model organisms such as the developing zebrafish, are also part of the testing strategy. A variety of computational approaches are used to analyze the resulting complex data sets to gain insight in to inherent biological activity of chemicals and possible mechanisms of toxicity. Several case studies including identification of modulators of estrogen receptor and aromatic hydrocarbon receptor pathways with effects in primary human cell systems will be described. In addition, existing in vivo data from a subset of the chemicals was used to anchor predictive models using in vitro data for a number of adverse endpoints including reproductive and developmental toxicities. The strengths and weaknesses of this approach will be described. This work does not necessarily reflect official Agency policy. Pres

Formulation and process factors influencing product quality and in vitro performance of ophthalmic ointments.

Science.gov (United States)

Xu, Xiaoming; Al-Ghabeish, Manar; Rahman, Ziyaur; Krishnaiah, Yellela S R; Yerlikaya, Firat; Yang, Yang; Manda, Prashanth; Hunt, Robert L; Khan, Mansoor A

2015-09-30

Owing to its unique anatomical and physiological functions, ocular surface presents special challenges for both design and performance evaluation of the ophthalmic ointment drug products formulated with a variety of bases. The current investigation was carried out to understand and identify the appropriate in vitro methods suitable for quality and performance evaluation of ophthalmic ointment, and to study the effect of formulation and process variables on its critical quality attributes (CQA). The evaluated critical formulation variables include API initial size, drug percentage, and mineral oil percentage while the critical process parameters include mixing rate, temperature, time and cooling rate. The investigated quality and performance attributes include drug assay, content uniformity, API particle size in ointment, rheological characteristics, in vitro drug release and in vitro transcorneal drug permeation. Using design of experiments (DoE) as well as a novel principle component analysis approach, five of the quality and performance attributes (API particle size, storage modulus of ointment, high shear viscosity of ointment, in vitro drug release constant and in vitro transcorneal drug permeation rate constant) were found to be highly influenced by the formulation, in particular the strength of API, and to a lesser degree by processing variables. Correlating the ocular physiology with the physicochemical characteristics of acyclovir ophthalmic ointment suggested that in vitro quality metrics could be a valuable predictor of its in vivo performance. Published by Elsevier B.V.

Implementation challenges for designing Integrated In Vitro Testing Strategies (ITS) aiming at reducing and replacing animal experimentation

NARCIS (Netherlands)

Wever, B.de; Fuchs, H.W.; Gaca, M.; Krul, C.A.M.; Mikulowski, S.; Poth, A.; Roggen, E.L.; Vilà, M.R.

2012-01-01

At the IVTIP (In Vitro Testing Industrial Platform) meeting of November 26th 2009 entitled 'Toxicology in the 21st century ('21C') - working our way towards a visionary reality' all delegates endorsed the emerging concept of the '21C' vision as the way forward to enable a thorough, reliable and

Development of asthmatic inflammation in mice following early-life exposure to ambient environmental particulates and chronic allergen challenge

Directory of Open Access Journals (Sweden)

Cristan Herbert

2013-03-01

Childhood exposure to environmental particulates increases the risk of development of asthma. The underlying mechanisms might include oxidant injury to airway epithelial cells (AEC. We investigated the ability of ambient environmental particulates to contribute to sensitization via the airways, and thus to the pathogenesis of childhood asthma. To do so, we devised a novel model in which weanling BALB/c mice were exposed to both ambient particulate pollutants and ovalbumin for sensitization via the respiratory tract, followed by chronic inhalational challenge with a low mass concentration of the antigen. We also examined whether these particulates caused oxidant injury and activation of AEC in vitro. Furthermore, we assessed the potential benefit of minimizing oxidative stress to AEC through the period of sensitization and challenge by dietary intervention. We found that characteristic features of asthmatic inflammation developed only in animals that received particulates at the same time as respiratory sensitization, and were then chronically challenged with allergen. However, these animals did not develop airway hyper-responsiveness. Ambient particulates induced epithelial injury in vitro, with evidence of oxidative stress and production of both pro-inflammatory cytokines and Th2-promoting cytokines such as IL-33. Treatment of AEC with an antioxidant in vitro inhibited the pro-inflammatory cytokine response to these particulates. Ambient particulates also induced pro-inflammatory cytokine expression following administration to weanling mice. However, early-life dietary supplementation with antioxidants did not prevent the development of an asthmatic inflammatory response in animals that were exposed to particulates, sensitized and challenged. We conclude that injury to airway epithelium by ambient environmental particulates in early life is capable of promoting the development of an asthmatic inflammatory response in sensitized and antigen-challenged mice. These

Review of microfluidic cell culture devices for the control of gaseous microenvironments in vitro

Science.gov (United States)

Wu, H.-M.; Lee, T.-A.; Ko, P.-L.; Chiang, H.-J.; Peng, C.-C.; Tung, Y.-C.

2018-04-01

Gaseous microenvironments play important roles in various biological activities in vivo. However, it is challenging to precisely control gaseous microenvironments in vitro for cell culture due to the high diffusivity nature of gases. In recent years, microfluidics has paved the way for the development of new types of cell culture devices capable of manipulating cellular microenvironments, and provides a powerful tool for in vitro cell studies. This paper reviews recent developments of microfluidic cell culture devices for the control of gaseous microenvironments, and discusses the advantages and limitations of current devices. We conclude with suggestions for the future development of microfluidic cell culture devices for the control of gaseous microenvironments.

Molecular Biosensors for Electrochemical Detection of Infectious Pathogens in Liquid Biopsies: Current Trends and Challenges.

Science.gov (United States)

Campuzano, Susana; Yáñez-Sedeño, Paloma; Pingarrón, José Manuel

2017-11-03

Rapid and reliable diagnosis of infectious diseases caused by pathogens, and timely initiation of appropriate treatment are critical determinants to promote optimal clinical outcomes and general public health. Conventional in vitro diagnostics for infectious diseases are time-consuming and require centralized laboratories, experienced personnel and bulky equipment. Recent advances in electrochemical affinity biosensors have demonstrated to surpass conventional standards in regards to time, simplicity, accuracy and cost in this field. The tremendous potential offered by electrochemical affinity biosensors to detect on-site infectious pathogens at clinically relevant levels in scarcely treated body fluids is clearly stated in this review. The development and application of selected examples using different specific receptors, assay formats and electrochemical approaches focusing on the determination of specific circulating biomarkers of different molecular (genetic, regulatory and functional) levels associated with bacterial and viral pathogens are critically discussed. Existing challenges still to be addressed and future directions in this rapidly advancing and highly interesting field are also briefly pointed out.

Challenges of analysing suspected over exposed subjects using biological dosimetry at Sri Ramachandra University

International Nuclear Information System (INIS)

Vijayalakshimi, J.; Venkatachalam, P.; Solomon, F.D. Paul

2016-01-01

Biological dosimetry based on the analysis of dicentric chromosomes has become a routine component of the radiological protection programmes and has a valuable role to contribute in suspected over exposed subjects who perform diagnostic and therapeutic procedures. The Department of Human Genetics, Sri Ramachandra University, Porur, Chennai, has been involved in the standardization of chromosomal aberration analysis as a biological dosimeter for investigating accidental ionising radiation exposure since 1998. Our laboratory has been accredited since 2007 by Atomic Energy Regulatory Board. The initial process was to establish the in vitro dose response curve for various type of low LET ionizing radiation. Since accreditation, a total of 61 subjects have been referred to Sri Ramachandra University from SRRC, Kalpakkam. Brief social/medical history and informed consent are being obtained prior to blood samplings. The dose estimates expressed in sievert (Sv) measured by Thermoluminescence badges was in the range of 0.05-2779.05 mSv. Chromosomal aberration assay was used for analysis which allows direct detection of aberration in peripheral blood lymphocytes. The test was performed as per the standard operating protocol on peripheral blood lymphocyte. Currently the dose response curve for the automated scoring process in under way and we hope to improve upon quality and turnaround time using the automation available. Future challenge would be to establish an in vitro dose response curve with automated scoring technique and developing inter-laboratory comparison of dose response generated using automation

Direct recovery of infectious Pestivirus from a full-length RT-PCR amplicon

DEFF Research Database (Denmark)

Rasmussen, Thomas Bruun; Reimann, Ilona; Hoffmann, Bernd

2008-01-01

This study describes the use of a novel and rapid long reverse transcription (RT)-PCR for the generation of infectious full-length cDNA of pestiviruses. To produce rescued viruses, full-length RT-PCR amplicons of 12.3 kb, including a T7-promotor, were transcribed directly in vitro, and the result......This study describes the use of a novel and rapid long reverse transcription (RT)-PCR for the generation of infectious full-length cDNA of pestiviruses. To produce rescued viruses, full-length RT-PCR amplicons of 12.3 kb, including a T7-promotor, were transcribed directly in vitro......, and the resulting RNA transcripts were electroporated into ovine cells. Infectious virus was obtained after one cell culture passage. The rescued viruses had a phenotype similar to the parental Border Disease virus strain. Therefore, direct generation of infectious pestiviruses from full-length RT-PCR cDNA products...

In vitro root induction of faba bean (Vicia faba L.).

Science.gov (United States)

Ismail, Roba M; Elazab, Heba E M; Hussein, Gihan M H; Metry, Emad A

2011-01-01

A major challenge for regeneration of faba bean (Vicia faba L.) plants is the difficulty of in vitro root induction. In the present study, in vitro rooting and its architecture have been studied. Adventitious root formation was successfully induced from regenerated faba bean shoots of four Egyptian cultivars, i.e., Giza 461, Giza 40, Giza 834 and Giza 716 on hormone free MS medium supplemented with 5 mg/l silver nitrate. Among the four cultivars, Giza 461 and Giza 40 were recorded as the highest root formation response (75 % and 65) followed by cultivars Giza716 and Giza843 (20%, and 10%). Anatomical study proved that the produced roots are initiated as the adventitious lateral root (LR) with tri-arch xylem strands as compared with the penta-arch of the primary roots of the intact faba bean seedling. The obtained results overcome the root induction problem in faba bean.

In Vitro Analysis of Metabolite Transport Proteins.

Science.gov (United States)

Roell, Marc-Sven; Kuhnert, Franziska; Zamani-Nour, Shirin; Weber, Andreas P M

2017-01-01

The photorespiratory cycle is distributed over four cellular compartments, the chloroplast, peroxisomes, cytoplasm, and mitochondria. Shuttling of photorespiratory intermediates between these compartments is essential to maintain the function of photorespiration. Specific transport proteins mediate the transport across biological membranes and represent important components of the cellular metabolism. Although significant progress was made in the last years on identifying and characterizing new transport proteins, the overall picture of intracellular metabolite transporters is still rather incomplete. The photorespiratory cycle requires at least 25 transmembrane transport steps; however to date only plastidic glycolate/glycerate transporter and the accessory 2-oxoglutarate/malate and glutamate/malate transporters as well as the mitochondrial transporter BOU1 have been identified. The characterization of transport proteins and defining their substrates and kinetics are still major challenges.Here we present a detailed set of protocols for the in vitro characterization of transport proteins. We provide protocols for the isolation of recombinant transport protein expressed in E. coli or Saccharomyces cerevisiae and the extraction of total leaf membrane protein for in vitro analysis of transporter proteins. Further we explain the process of reconstituting transport proteins in artificial lipid vesicles and elucidate the details of transport assays.

Design, Results and Plans for Power Beaming Competitive Challenge

International Nuclear Information System (INIS)

Shelef, Ben

2008-01-01

In our context, Power Beaming refers to the extraction of useable electrical power from a directed electromagnetic beam. In order to promote interest in this technology, the Spaceward Foundation proposed and is managing a technology prize challenge based on a Space Elevator design scenario. The challenge has a prize purse of $2M, provided by NASA's Centennial Challenges office. This paper covers the considerations that went into the design of the challenge, a brief chronology of past results, and plans for the future

In vitro selection of shape-changing DNA nanostructures capable of binding-induced cargo release.

Science.gov (United States)

Oh, Seung Soo; Plakos, Kory; Xiao, Yi; Eisenstein, Michael; Soh, H Tom

2013-11-26

Many biological systems employ allosteric regulatory mechanisms, which offer a powerful means of directly linking a specific binding event to a wide spectrum of molecular functionalities. There is considerable interest in generating synthetic allosteric regulators that can perform useful molecular functions for applications in diagnostics, imaging and targeted therapies, but generating such molecules through either rational design or directed evolution has proven exceptionally challenging. To address this need, we present an in vitro selection strategy for generating conformation-switching DNA nanostructures that selectively release a small-molecule payload in response to binding of a specific trigger molecule. As an exemplar, we have generated a DNA nanostructure that hybridizes with a separate 'cargo strand' containing an abasic site. This abasic site stably sequesters a fluorescent cargo molecule in an inactive state until the DNA nanostructure encounters an ATP trigger molecule. This ATP trigger causes the nanostructure to release the cargo strand, thereby liberating the fluorescent payload and generating a detectable fluorescent readout. Our DNA nanostructure is highly sensitive, with an EC50 of 30 μM, and highly specific, releasing its payload in response to ATP but not to other chemically similar nucleotide triphosphates. We believe that this selection approach could be generalized to generate synthetic nanostructures capable of selective and controlled release of other small-molecule cargos in response to a variety of triggers, for both research and clinical applications.

Glycosaminoglycan-based hydrogels to modulate heterocellular communication in in vitro angiogenesis models

Science.gov (United States)

Chwalek, Karolina; Tsurkan, Mikhail V.; Freudenberg, Uwe; Werner, Carsten

2014-03-01

Angiogenesis, the outgrowth of blood vessels, is crucial in development, disease and regeneration. Studying angiogenesis in vitro remains challenging because the capillary morphogenesis of endothelial cells (ECs) is controlled by multiple exogenous signals. Therefore, a set of in situ-forming starPEG-heparin hydrogels was used to identify matrix parameters and cellular interactions that best support EC morphogenesis. We showed that a particular type of soft, matrix metalloproteinase-degradable hydrogel containing covalently bound integrin ligands and reversibly conjugated pro-angiogenic growth factors could boost the development of highly branched, interconnected, and lumenized endothelial capillary networks. Using these effective matrix conditions, 3D heterocellular interactions of ECs with different mural cells were demonstrated that enabled EC network modulation and maintenance of stable vascular capillaries over periods of about one month in vitro. The approach was also shown to permit in vitro tumor vascularization experiments with unprecedented levels of control over both ECs and tumor cells. In total, the introduced 3D hydrogel co-culture system could offer unique options for dissecting and adjusting biochemical, biophysical, and cell-cell triggers in tissue-related vascularization models.

Nutraceuticals, A New Challenge for Medicinal Chemistry.

Science.gov (United States)

Sut, Stefania; Baldan, Valeria; Faggian, Marta; Peron, Gregorio; Dall Acqua, Stefano

2016-01-01

"Nutraceuticals" are food-derived products largely used for their presumed healthpromoting or disease-preventing effects. In the recent years, many efforts have been aimed at assessing nutraceutical efficacy and safety, but these factors are difficult to address because of the complex chemical compositions and multiple mode of actions. Thus, the study of nutraceutical ingredients poses several challenges for the medicinal chemistry field, some of which are related to extraction and chemical characterization, some to in vitro and in vivo bioactivity evaluation, and some to the bioavailability and interaction of these natural mixtures with organs and microbiota. Furthermore, because of their nature as medicinal and food products, these nutraceuticals can also be considered as a valuable source of new "lead compounds", creating the opportunity to discover new classes of therapeutic agents. This review provides information on these themes, showing the new challenges that comprehensive medicinal chemistry research is called to answer in the field of nutraceuticals.

New directions: Air pollution challenges for developing megacities like Delhi

Science.gov (United States)

Kumar, Prashant; Khare, Mukesh; Harrison, Roy M.; Bloss, William J.; Lewis, Alastair C.; Coe, Hugh; Morawska, Lidia

2015-12-01

Most major cities around the world experience periods of elevated air pollution levels, which exceed international health-based air quality standards (Kumar et al., 2013). Although it is a global problem, some of the highest air pollution levels are found in rapidly expanding cities in India and China. The sources, emissions, transformations and broad effects of meteorology on air pollution are reasonably well accounted in air quality control strategies in many developed cities; however these key factors remain poorly constrained in the growing cities of countries with emerging economies. We focus here on Delhi, one of the largest global population centres, which faces particular air pollution challenges, now and in the future.

Gaining acceptance for the use of in vitro toxicity assays and QIVIVE in regulatory risk assessment.

Science.gov (United States)

Meek, M E Bette; Lipscomb, John C

2015-06-05

Testing strategies are anticipated to increasingly rely on in vitro data as a basis to characterize early steps or key events in toxicity at relevant dose levels in human tissues. Such strategies require quantitative in vitro to in vivo extrapolation to characterize dose-response as a basis for comparison with exposure to estimate risk. Current experience in the incorporation of mechanistic and in vitro data in risk assessment is considered here in the context of identified principles to increase the potential for timely acceptance of more progressive and tailored testing strategies by the regulatory community. These principles are outlined as transitioning in a familiar context, tiering to acquire experience and increase confidence, contextual knowledge transfer to facilitate interpretation and communication, coordination and development of expertise and continuing challenge. A proposed pragmatic tiered data driven framework which includes increasing reliance on in vitro data and quantitative in vitro to in vivo extrapolation is considered in the context of these principles. Based on this analysis, possible additional steps that might facilitate timely evolution and potentially, uptake are identified. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

ENRAIZAMENTO DE CRAVO (Dianthus caryophyllus L. IN VITRO E EX VITRO

Directory of Open Access Journals (Sweden)

G.R.F CUZZUOL

1996-01-01

Full Text Available Plântulas de cravo (Dianthus caryophyllus micropropagadas durante várias gerações pelo período de um ano, foram enraizadas in vitro com AIA, ANA e AIB nas concentrações de 0,0; 0,25; 0,5 e 1,0 mg/l, em fatorial do tipo 3 x 4, com todos os tratamentos promovendo a formação de raízes, mas não diferindo do controle. Foi confrontado em condição autotrófica, o desempenho entre plântulas enraizadas in vitro na presença e ausência do regulador AIA 0,5 mg/l e plântulas enraizadas ex vitro, sem nenhuma diferença quanto ao comprimento da parte aérea. Para a variável produção de massa de matéria seca os melhores resultados foram proporcionados pelas plantas que passaram pela fase de enraizamento in vitro, tendo o sistema radicular efeito sinergístico no crescimento da parte aérea.Plantlets of carnation (Dianthus caryophyllus L. micropropagated through several generations during one year, were observed with respect to rooting in vitro, in the presence of IAA , NAA and IBA, at the following concentrations: 0,0; 0,25; 0,5 and 1,0 mg/l. All treatments promoted root formation, however no differences were detected in comparison to control. As far as the lenght of the aerial part is concerned no difference was observed between in vitro rooting. in the presence or absence of IAA 0,5 mg/l, and ex vitro rooting. Plantlets which were rooted in vitro conditions showed higher production of fresh matter then those rooted ex vitro. The root system had a synergistic effect on the growth of the aerial part.

Characterisation of neutron fields: challenges in assessing the directional distribution

International Nuclear Information System (INIS)

Cauwels, Vanessa; Vanhavere, Filip; Reginatto, Marcel

2014-01-01

The SCK.CEN has carried out neutron field characterisation campaigns at several nuclear reactors. The main goal of these measurement campaigns was to evaluate the performance of different neutron personal dosemeters. To be able to evaluate the performance of neutron personal dosemeters in terms of H p (10), knowledge of the directional distribution is indispensable. This distribution was estimated by placing several personal dosemeters on all six sides of a slab phantom. The interpretation and conversion of this information into a reliable value for H p (10) requires great care. The data were analysed using three methods. In the first approach, a linear interpolation was performed on three perpendicular axes. In the other two approaches, an icosahedron was used to model the angle of incidence of the neutrons and a linear interpolation or a Bayesian analysis was performed. This study describes the limitations and advantages of each of these methods and provides recommendations for their use to estimate the personal dose equivalent H p (10) for neutron dosimetry. Neutron personal dosimetry is complicated by the fact that the neutron dose quantity H p (10) is strongly energy and angular dependent. Instead of simply assuming that the fluence is unidirectional or that the fluence is isotropic, an attempt was made to estimate the directional distribution of the neutron field using a relatively simple measurement procedure. A number of active and passive personal dosemeters were placed on the six faces of a slab phantom and the results were analysed via different algorithms to obtain partial fluences in several directions of incidence. The results from all calculations in this study show the importance of introducing information about the directional distribution of the neutron fluence for the estimation of the personal dose equivalent H p (10). The difference between H p (10) dose estimates carried out using a unidirectional or an isotropic distribution can be of up

In vitro irradiation treatment - an effective method of breeding for chrysanthemum

International Nuclear Information System (INIS)

Dao Thanh Bang; Nguyen Hong Nhung; Nguyen Phuong Doai; Le Thi Lieu; Nguyen Pham Hung

2011-01-01

Recently, mutation induction is one of the an effective tool for crop breeding improvement. However, challenge for breeders is how to find suitable method for vegetative crop. Strong point of in vitro mutation breeding is reduce breeding process, easy to remove chimeric mutant after several cycle multiplication tissue culture. Based on optimal media of callus formation, regeneration and root media in chrysanthemum.The experiment of of irradiation treatment was carried out at the range doses 10, 20, 30, 50 and 70 Gy for callus of bud. From generation of M1V4, promising mutants was selected mostly in the dose of 30 Gy and bring back to in vitro for fixation and multiplication of mutant lines. Selection and evaluation of mutant lines was carried at M1V8. From CN43, crystal yellow and Taiwan purple origin varieties we received three mutants VCM1, VCM2 and VCM3 respectively. All mutant varieties have been certified as regional varieties according to decision of Ministry of Agriculture and Rural Development. The result of research show that combination between in vitro irradiation and tissue culture is effective solution for chrysanthemum breeding in particular and vegetative crop in general. (author)

Direct PCR amplification of forensic touch and other challenging DNA samples: A review.

Science.gov (United States)

Cavanaugh, Sarah E; Bathrick, Abigail S

2018-01-01

DNA evidence sample processing typically involves DNA extraction, quantification, and STR amplification; however, DNA loss can occur at both the DNA extraction and quantification steps, which is not ideal for forensic evidence containing low levels of DNA. Direct PCR amplification of forensic unknown samples has been suggested as a means to circumvent extraction and quantification, thereby retaining the DNA typically lost during those procedures. Direct PCR amplification is a method in which a sample is added directly to an amplification reaction without being subjected to prior DNA extraction, purification, or quantification. It allows for maximum quantities of DNA to be targeted, minimizes opportunities for error and contamination, and reduces the time and monetary resources required to process samples, although data analysis may take longer as the increased DNA detection sensitivity of direct PCR may lead to more instances of complex mixtures. ISO 17025 accredited laboratories have successfully implemented direct PCR for limited purposes (e.g., high-throughput databanking analysis), and recent studies indicate that direct PCR can be an effective method for processing low-yield evidence samples. Despite its benefits, direct PCR has yet to be widely implemented across laboratories for the processing of evidentiary items. While forensic DNA laboratories are always interested in new methods that will maximize the quantity and quality of genetic information obtained from evidentiary items, there is often a lag between the advent of useful methodologies and their integration into laboratories. Delayed implementation of direct PCR of evidentiary items can be attributed to a variety of factors, including regulatory guidelines that prevent laboratories from omitting the quantification step when processing forensic unknown samples, as is the case in the United States, and, more broadly, a reluctance to validate a technique that is not widely used for evidence samples. The

Sustained Release of a Water-Soluble Drug from Directly ...

African Journals Online (AJOL)

Okra gum was evaluated as a controlled-release agent in modified release matrices in comparison with sodium carboxymethylcellulose (NaCMC) using aspirin as the model drug. Tablets were produced by direct compression and the in vitro drug release was assessed under conditions similar to those in the gastrointestinal ...

In vitro biocompatibility of Ti-Mg alloys fabricated by direct current magnetron sputtering.

Science.gov (United States)

Hieda, Junko; Niinomi, Mitsuo; Nakai, Masaaki; Cho, Ken

2015-09-01

Ti-xMg (x=17, 33, and 55 mass%) alloy films, which cannot be prepared by conventional melting processes owing to the absence of a solid-solution phase in the phase diagram, were prepared by direct current magnetron sputtering in order to investigate their biocompatibility. Ti and Mg films were also prepared by the same process for comparison. The crystal structures were examined by X-ray diffraction (XRD) analysis and the surfaces were analyzed by X-ray photoelectron spectroscopy. The Ti, Ti-xMg alloy, and Mg films were immersed in a 0.9% NaCl solution at 310 K for 7d to evaluate the dissolution amounts of Ti and Mg. In addition, to evaluate the formation ability of calcium phosphate in vitro, the Ti, Ti-xMg alloy, and Mg films were immersed in Hanks' solution at 310 K for 30 d. Ti and Mg form solid-solution alloys because the peaks attributed to pure Ti and Mg do not appear in the XRD patterns of any of the Ti-xMg alloy films. The surfaces of the Ti-17 Mg alloy and Ti-33 Mg alloy films contain Ti oxides and MgO, whereas MgO is the main component of the surface oxide of the Ti-55 Mg alloy and Mg films. The dissolution amounts of Ti from all films are below or near the detection limit of inductively coupled plasma-optical emission spectroscopy. On the other hand, the Ti-17 Mg alloy, Ti-33 Mg alloy, Ti-55 Mg alloy, and Mg films exhibit Mg dissolution amounts of approximately 2.5, 1.4, 21, and 41 μg/cm(2), respectively. The diffraction peaks attributed to calcium phosphate are present in the XRD patterns of the Ti-33 Mg alloy, Ti-55 Mg alloy, and Mg films after the immersion in Hanks' solution. Spherical calcium phosphate particles precipitate on the surface of the Ti-33 Mg film. However, many cracks are observed in the Ti-55 Mg film, and delamination of the film occurs after the immersion in Hanks' solution. The Mg film is dissolved in Hanks' solution and calcium phosphate particles precipitate on the glass substrate. Consequently, it is revealed that the Ti-33 Mg

Acute in vitro and in vivo toxicity of a commercial grade boron nitride nanotube mixture.

Science.gov (United States)

Kodali, Vamsi K; Roberts, Jenny R; Shoeb, Mohammad; Wolfarth, Michael G; Bishop, Lindsey; Eye, Tracy; Barger, Mark; Roach, Katherine A; Friend, Sherri; Schwegler-Berry, Diane; Chen, Bean T; Stefaniak, Aleksandr; Jordan, Kevin C; Whitney, Roy R; Porter, Dale W; Erdely, Aaron D

2017-10-01

Boron nitride nanotubes (BNNTs) are an emerging engineered nanomaterial attracting significant attention due to superior electrical, chemical and thermal properties. Currently, the toxicity profile of this material is largely unknown. Commercial grade BNNTs are composed of a mixture (BNNT-M) of ∼50-60% BNNTs, and ∼40-50% impurities of boron and hexagonal boron nitride. We performed acute in vitro and in vivo studies with commercial grade BNNT-M, dispersed by sonication in vehicle, in comparison to the extensively studied multiwalled carbon nanotube-7 (MWCNT-7). THP-1 wild-type and NLRP3-deficient human monocytic cells were exposed to 0-100 µg/ml and C57BL/6 J male mice were treated with 40 µg of BNNT-M for in vitro and in vivo studies, respectively. In vitro, BNNT-M induced a dose-dependent increase in cytotoxicity and oxidative stress. This was confirmed in vivo following acute exposure increase in bronchoalveolar lavage levels of lactate dehydrogenase, pulmonary polymorphonuclear cell influx, loss in mitochondrial membrane potential and augmented levels of 4-hydroxynonenal. Uptake of this material caused lysosomal destabilization, pyroptosis and inflammasome activation, corroborated by an increase in cathepsin B, caspase 1, increased protein levels of IL-1β and IL-18 both in vitro and in vivo. Attenuation of these effects in NLRP3-deficient THP-1 cells confirmed NLRP3-dependent inflammasome activation by BNNT-M. BNNT-M induced a similar profile of inflammatory pulmonary protein production when compared to MWCNT-7. Functionally, pretreatment with BNNT-M caused suppression in bacterial uptake by THP-1 cells, an effect that was mirrored in challenged alveolar macrophages collected from exposed mice and attenuated with NLRP3 deficiency. Analysis of cytokines secreted by LPS-challenged alveolar macrophages collected after in vivo exposure to dispersions of BNNT-M showed a differential macrophage response. The observed results demonstrated acute

Stigma and Schizophrenia: Directions in Student Training

Science.gov (United States)

Mason, Susan E.; Miller, Rachel

2006-01-01

The persistence of stigma related to schizophrenia is addressed as a continuing challenge for social work students working in mental health. Student education is best grounded in direct clinical work with clients in conjunction with field supervision and classroom education. Through direct practice in individual and group sessions, students learn…

Simulation of lung alveolar epithelial wound healing in vitro.

Science.gov (United States)

Kim, Sean H J; Matthay, Michael A; Mostov, Keith; Hunt, C Anthony

2010-08-06

The mechanisms that enable and regulate alveolar type II (AT II) epithelial cell wound healing in vitro and in vivo remain largely unknown and need further elucidation. We used an in silico AT II cell-mimetic analogue to explore and better understand plausible wound healing mechanisms for two conditions: cyst repair in three-dimensional cultures and monolayer wound healing. Starting with the analogue that validated for key features of AT II cystogenesis in vitro, we devised an additional cell rearrangement action enabling cyst repair. Monolayer repair was enabled by providing 'cells' a control mechanism to switch automatically to a repair mode in the presence of a distress signal. In cyst wound simulations, the revised analogue closed wounds by adhering to essentially the same axioms available for alveolar-like cystogenesis. In silico cell proliferation was not needed. The analogue recovered within a few simulation cycles but required a longer recovery time for larger or multiple wounds. In simulated monolayer wound repair, diffusive factor-mediated 'cell' migration led to repair patterns comparable to those of in vitro cultures exposed to different growth factors. Simulations predicted directional cell locomotion to be critical for successful in vitro wound repair. We anticipate that with further use and refinement, the methods used will develop as a rigorous, extensible means of unravelling mechanisms of lung alveolar repair and regeneration.

Low immunogenicity of allogeneic human umbilical cord blood-derived mesenchymal stem cells in vitro and in vivo

International Nuclear Information System (INIS)

Lee, Miyoung; Jeong, Sang Young; Ha, Jueun; Kim, Miyeon; Jin, Hye Jin; Kwon, Soon-Jae; Chang, Jong Wook; Choi, Soo Jin; Oh, Wonil; Yang, Yoon Sun; Kim, Jae-Sung; Jeon, Hong Bae

2014-01-01

Highlights: • hUCB-MSCs maintained low immunogenicity even after immune challenge in vitro. • Humanized NSG mice were established using human UCB CD34+ cells. • Repeated intravenous hUCB-MSC injection into mice did not lead to immune responses and adverse events. • Allogeneic hUCB-MSCs maintained low immunogenicity in vitro and in vivo. - Abstract: Evaluation of the immunogenicity of human mesenchymal stem cells (MSCs) in an allogeneic setting during therapy has been hampered by lack of suitable models due to technical and ethical limitations. Here, we show that allogeneic human umbilical cord blood derived-MSCs (hUCB-MSCs) maintained low immunogenicity even after immune challenge in vitro. To confirm these properties in vivo, a humanized mouse model was established by injecting isolated hUCB-derived CD34+ cells intravenously into immunocompromised NOD/SCID IL2γnull (NSG) mice. After repeated intravenous injection of human peripheral blood mononuclear cells (hPBMCs) or MRC5 cells into these mice, immunological alterations including T cell proliferation and increased IFN-γ, TNF-α, and human IgG levels, were observed. In contrast, hUCB-MSC injection did not elicit these responses. While lymphocyte infiltration in the lung and small intestine and reduced survival rates were observed after hPBMC or MRC5 transplantation, no adverse events were observed following hUCB-MSC introduction. In conclusion, our data suggest that allogeneic hUCB-MSCs have low immunogenicity in vitro and in vivo, and are therefore “immunologically safe” for use in allogeneic clinical applications

Low immunogenicity of allogeneic human umbilical cord blood-derived mesenchymal stem cells in vitro and in vivo

Energy Technology Data Exchange (ETDEWEB)

Lee, Miyoung; Jeong, Sang Young; Ha, Jueun; Kim, Miyeon; Jin, Hye Jin; Kwon, Soon-Jae [Biomedical Research Institute, MEDIPOST Co., Ltd, Seoul 137-874 (Korea, Republic of); Chang, Jong Wook [Research Institute for Future Medicine Stem Cell and Regenerative Medicine Center, Samsung Medical Center, Seoul 137-710 (Korea, Republic of); Choi, Soo Jin; Oh, Wonil; Yang, Yoon Sun [Biomedical Research Institute, MEDIPOST Co., Ltd, Seoul 137-874 (Korea, Republic of); Kim, Jae-Sung [Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, Seoul 139-709 (Korea, Republic of); Jeon, Hong Bae, E-mail: jhb@medi-post.co.kr [Biomedical Research Institute, MEDIPOST Co., Ltd, Seoul 137-874 (Korea, Republic of)

2014-04-18

Highlights: • hUCB-MSCs maintained low immunogenicity even after immune challenge in vitro. • Humanized NSG mice were established using human UCB CD34+ cells. • Repeated intravenous hUCB-MSC injection into mice did not lead to immune responses and adverse events. • Allogeneic hUCB-MSCs maintained low immunogenicity in vitro and in vivo. - Abstract: Evaluation of the immunogenicity of human mesenchymal stem cells (MSCs) in an allogeneic setting during therapy has been hampered by lack of suitable models due to technical and ethical limitations. Here, we show that allogeneic human umbilical cord blood derived-MSCs (hUCB-MSCs) maintained low immunogenicity even after immune challenge in vitro. To confirm these properties in vivo, a humanized mouse model was established by injecting isolated hUCB-derived CD34+ cells intravenously into immunocompromised NOD/SCID IL2γnull (NSG) mice. After repeated intravenous injection of human peripheral blood mononuclear cells (hPBMCs) or MRC5 cells into these mice, immunological alterations including T cell proliferation and increased IFN-γ, TNF-α, and human IgG levels, were observed. In contrast, hUCB-MSC injection did not elicit these responses. While lymphocyte infiltration in the lung and small intestine and reduced survival rates were observed after hPBMC or MRC5 transplantation, no adverse events were observed following hUCB-MSC introduction. In conclusion, our data suggest that allogeneic hUCB-MSCs have low immunogenicity in vitro and in vivo, and are therefore “immunologically safe” for use in allogeneic clinical applications.

Anatomia comparada das folhas e raízes de Cymbidium Hort. (Orchidaceae cultivadas ex vitro e in vitro Comparative leaf and root anatomy of ex vitro and in vitro cultured Cymbidium Hort. plants

Directory of Open Access Journals (Sweden)

Juliana Lischka Sampaio Mayer

2008-06-01

Full Text Available Na fase de cultivo in vitro, as plantas são mantidas em ambiente com alta umidade relativa do ar, baixa luminosidade e trocas gasosas restritas, o que resulta em taxa de transpiração reduzida. Portanto, quando essas mudas são expostas ao meio ex vitro, sofrem estresse que pode causar a morte. O objetivo desse trabalho foi comparar a estrutura anatômica das mudas de Cymbidium 'Joy Polis' cultivadas ex vitro (planta matriz e aclimatizada e in vitro e verificar se a estrutura anatômica das plantas in vitro influencia no processo de aclimatização. As plantas ex vitro foram mantidas em casa-de-vegetação, em vasos individuais com o substrato fibra de coco em pó combinada com fibra de coco, e as plantas in vitro foram mantidas em meio de cultura MS. Para a análise anatômica qualitativa foram coletadas amostras de folhas e raízes de plantas ex vitro e in vitro. As plantas aclimatizadas apresentaram estrutura morfoanatômica semelhante à da planta matriz. A estrutura anatômica das plantas in vitro não influenciou a sobrevivência das mudas durante a aclimatização devido à plasticidade fenotipica desse cultivar. As plantas de Cymbidium 'Joy Polis' possuem grande capacidade de aclimatização ao ambiente, sendo provavelmente este um dos fatores responsáveis pela sobrevivência de 100% das mudas.During in vitro culture plants are kept in an atmosphere with high relative humidity, low light intensity and reduced gas exchange, resulting in low transpiration rates. Therefore, when these plants are exposed to ex vitro conditions, they suffer stress, which can induce mortality. The purpose of this study was to compare the anatomical structure of Cymbidium 'Joy Polis' plants from ex vitro (mother plant and acclimatized plants and in vitro cultures and to verify if the anatomical structure of in vitro cultured plants affects acclimatization. The ex vitro plants were kept in a greenhouse in pots containing a mixture of coconut-fiber powder and

Influence of chlorhexidine and/or ethanol treatment on bond strength of an etch-and-rinse adhesive to dentin: an in vitro and in situ study.

Science.gov (United States)

Simões, D M S; Basting, R T; Amaral, F L B; Turssi, C P; França, F M G

2014-01-01

The aim of this study was to evaluate the effect of a chlorhexidine and/or ethanol application on the bond strength of an etch-and-rinse, hydrophobic adhesive system either under in vitro aging or in situ cariogenic challenge. The dentin surface of 36 human third molars were flattened and allocated into four groups to be treated with chlorhexidine, ethanol, or chlorhexidine + ethanol or left unexposed to any solution (control) (n=9). Then, a resin composite restoration was made on the dentin surface and longitudinal sticks were obtained. Sticks from each tooth were assigned to three test conditions: stored in water in vitro for 24 hours, stored in water in vitro for 6 months, or worn in situ for 14 days. During in situ wear time, a high-cariogenic challenge condition was simulated. Specimens were tested for microtensile bond strength (μTBS). Multivariate analysis of variance and Tukey's test showed that chlorhexidine, ethanol, or chlorhexidine + ethanol did not affect the μTBS. The in vitro μTBS values were significantly lower for the specimens stored for 6 months than for those stored for 24 hours. Intermediate μTBS values were shown by the specimens worn in situ. Thus, use of chlorhexidine and/or ethanol was incapable of containing the degradation at the bond interface in the in vitro model. The in situ model was capable of reducing bond strength similarly to the in vitro/6 months model. Despite this, the in situ bond strength was still similar to that of the in vitro/24-hour model.

The secretory products of Trichomonas vaginalis decrease fertilizing capacity of mice sperm in vitro

Science.gov (United States)

Roh, Jaesook; Lim, Young-Su; Seo, Min-Young; Choi, Yuri; Ryu, Jae-Sook

2015-01-01

Trichomonas vaginalis infection is one of the most prevalent sexually transmitted infections in humans and is now recognized as an important cause of infertility in men. There is little information about the effect of extracellular polymeric substances (EPS) from T. vaginalis on sperm, but previous reports do not provide a conclusive description of the functional integrity of the sperm. To investigate the impact of EPS on the fertilizing capacity of sperm, we assessed sperm motility, acrosomal status, hypo-osmotic swelling, and in vitro fertilization rate after incubating the sperm with EPS in vitro using mice. The incubation of sperm with EPS significantly decreased sperm motility, viability, and functional integrity in a concentration and time-dependent manner. These effects on sperm quality also resulted in a decreased fertilization rate in vitro. This is the first report that demonstrates the direct negative impact of the EPS of T. vaginalis on the fertilization rate of sperm in vitro. However, further study should be performed using human sperm to determine if EPS has similar negative impact on human sperm fertilizing capacity in vitro. PMID:25578937

Special report: workshop on 4D-treatment planning in actively scanned particle therapy--recommendations, technical challenges, and future research directions.

Science.gov (United States)

Knopf, Antje; Bert, Christoph; Heath, Emily; Nill, Simeon; Kraus, Kim; Richter, Daniel; Hug, Eugen; Pedroni, Eros; Safai, Sairos; Albertini, Francesca; Zenklusen, Silvan; Boye, Dirk; Söhn, Matthias; Soukup, Martin; Sobotta, Benjamin; Lomax, Antony

2010-09-01

This article reports on a 4D-treatment planning workshop (4DTPW), held on 7-8 December 2009 at the Paul Scherrer Institut (PSI) in Villigen, Switzerland. The participants were all members of institutions actively involved in particle therapy delivery and research. The purpose of the 4DTPW was to discuss current approaches, challenges, and future research directions in 4D-treatment planning in the context of actively scanned particle radiotherapy. Key aspects were addressed in plenary sessions, in which leaders of the field summarized the state-of-the-art. Each plenary session was followed by an extensive discussion. As a result, this article presents a summary of recommendations for the treatment of mobile targets (intrafractional changes) with actively scanned particles and a list of requirements to elaborate and apply these guidelines clinically.

Focus on the therapeutic efficacy of 3BNC117 against HIV-1: In vitro studies, in vivo studies, clinical trials and challenges.

Science.gov (United States)

Liu, Zhi-Jun; Bai, Jing; Liu, Feng-Li; Zhang, Xiang-Yang; Wang, Jing-Zhang

2017-11-01

3BNC117, which was discovered in 2011, is a broadly neutralizing antibody (bNAb) and specifically neutralizes the human immunodeficiency virus type-1 (HIV-1) by targeting the CD4-binding site. This is the first comprehensive review that focuses on the role of 3BNC117 in the prevention of HIV-1 and acquired immune deficiency syndrome (AIDS). Briefly, 3BNC117 neutralizes many HIV/SHIV strains in vitro, blocks HIV-1 acquisition in animal models via a pre-exposure prophylaxis, alleviates HIV-1-associated viremia via a post-exposure therapeutic effect, prevents the establishment of latent HIV-1 reservoirs, and induces both humoral and cellular anti-HIV immune responses in vivo. The outcomes of Phase I and Phase IIa clinical trials in 2015 and 2016 showed the safety, tolerability, and therapeutic efficacy of 3BNC117 in HIV-1-infected human individuals. Nevertheless, anti-3BNC117 antibodies and HIV-1 strains resistant to 3BNC117 pose clinical challenges to immunotherapy with 3BNC117, so potential strategies for optimizing the potency of 3BNC117 are suggested here. Predictably, HIV-1 prevention and AIDS treatment will benefit from combinational immunotherapies with 3BNC117 and other pharmaceuticals (bNAbs, antiretroviral medicines, viral inducers, etc.) in the near future. Copyright © 2017 Elsevier B.V. All rights reserved.

Macromolecular crowding directs extracellular matrix organization and mesenchymal stem cell behavior.

Directory of Open Access Journals (Sweden)

Adam S Zeiger

Full Text Available Microenvironments of biological cells are dominated in vivo by macromolecular crowding and resultant excluded volume effects. This feature is absent in dilute in vitro cell culture. Here, we induced macromolecular crowding in vitro by using synthetic macromolecular globules of nm-scale radius at physiological levels of fractional volume occupancy. We quantified the impact of induced crowding on the extracellular and intracellular protein organization of human mesenchymal stem cells (MSCs via immunocytochemistry, atomic force microscopy (AFM, and AFM-enabled nanoindentation. Macromolecular crowding in extracellular culture media directly induced supramolecular assembly and alignment of extracellular matrix proteins deposited by cells, which in turn increased alignment of the intracellular actin cytoskeleton. The resulting cell-matrix reciprocity further affected adhesion, proliferation, and migration behavior of MSCs. Macromolecular crowding can thus aid the design of more physiologically relevant in vitro studies and devices for MSCs and other cells, by increasing the fidelity between materials synthesized by cells in vivo and in vitro.

Macromolecular crowding directs extracellular matrix organization and mesenchymal stem cell behavior.

Science.gov (United States)

Zeiger, Adam S; Loe, Felicia C; Li, Ran; Raghunath, Michael; Van Vliet, Krystyn J

2012-01-01

Microenvironments of biological cells are dominated in vivo by macromolecular crowding and resultant excluded volume effects. This feature is absent in dilute in vitro cell culture. Here, we induced macromolecular crowding in vitro by using synthetic macromolecular globules of nm-scale radius at physiological levels of fractional volume occupancy. We quantified the impact of induced crowding on the extracellular and intracellular protein organization of human mesenchymal stem cells (MSCs) via immunocytochemistry, atomic force microscopy (AFM), and AFM-enabled nanoindentation. Macromolecular crowding in extracellular culture media directly induced supramolecular assembly and alignment of extracellular matrix proteins deposited by cells, which in turn increased alignment of the intracellular actin cytoskeleton. The resulting cell-matrix reciprocity further affected adhesion, proliferation, and migration behavior of MSCs. Macromolecular crowding can thus aid the design of more physiologically relevant in vitro studies and devices for MSCs and other cells, by increasing the fidelity between materials synthesized by cells in vivo and in vitro.

Privacy Challenges of Genomic Big Data.

Science.gov (United States)

Shen, Hong; Ma, Jian

2017-01-01

With the rapid advancement of high-throughput DNA sequencing technologies, genomics has become a big data discipline where large-scale genetic information of human individuals can be obtained efficiently with low cost. However, such massive amount of personal genomic data creates tremendous challenge for privacy, especially given the emergence of direct-to-consumer (DTC) industry that provides genetic testing services. Here we review the recent development in genomic big data and its implications on privacy. We also discuss the current dilemmas and future challenges of genomic privacy.

MicroRNA-494 inhibits cell proliferation and invasion of chondrosarcoma cells in vivo and in vitro by directly targeting SOX9.

Science.gov (United States)

Li, Jingyuan; Wang, Lijuan; Liu, Zongzhi; Zu, Chao; Xing, Fanfan; Yang, Pei; Yang, Yongkang; Dang, Xiaoqian; Wang, Kunzheng

2015-09-22

Accumulating evidence indicates that dysregulation of miRNAs could contribute to tumor growth and metastasis of chondrosarcoma by infuencing cell proliferation and invasion. In the current study, we are interested to examine the role of miRNAs in the carcinogenesis and progression of chondrosarcoma. Here, using comparative miRNA profiling of tissues and cells of chondrosarcoma and cartilage, we identified miR-494 as a commonly downregulated miRNA in the tissues of patients with chondrosarcoma and chondrosarcoma cancer cell line, and upregulation of miR-494 could inhibit proliferation and invasion of chondrosarcoma cancer cells in vivo and in vitro. Moreover, our data demonstrated that SOX9, the essential regulator of the process of cartilage differentiation, was the direct target and functional mediator of miR-494 in chondrosarcoma cells. And downregulation of SOX9 could also inhibit migration and invasion of chondrosarcoma cells. In the last, we identified low expression of miR-494 was significantly correlated with poor overall survival and prognosis of chondrosarcoma patients. Thus, miR-494 may be a new common therapeutic target and prognosis biomarker for chondrosarcoma.

Emotion regulation and mental health: recent findings, current challenges, and future directions.

Science.gov (United States)

Berking, Matthias; Wupperman, Peggilee

2012-03-01

In recent years, deficits in emotion regulation have been studied as a putative maintaining factor and promising treatment target in a broad range of mental disorders. This article aims to provide an integrative review of the latest theoretical and empirical developments in this rapidly growing field of research. Deficits in emotion regulation appear to be relevant to the development, maintenance, and treatment of various forms of psychopathology. Increasing evidence demonstrates that deficits in the ability to adaptively cope with challenging emotions are related to depression, borderline personality disorder, substance-use disorders, eating disorders, somatoform disorders, and a variety of other psychopathological symptoms. Unfortunately, studies differ with regard to the conceptualization and assessment of emotion regulation, thus limiting the ability to compare findings across studies. Future research should systematically work to use comparable methods in order to clarify the following: which individuals have; what kinds of emotion regulation difficulties with; which types of emotions; and what interventions are most effective in alleviating these difficulties. Despite some yet to be resolved challenges, the concept of emotion regulation has a broad and significant heuristic value for research in mental health.

In vivo and in vitro effects of selected antioxidants on rabbit meat microbiota.

Science.gov (United States)

Albonetti, Sabrina; Minardi, Paola; Trombetti, Fabiana; Savigni, Fabiana; Mordenti, Attilio Luigi; Baranzoni, Gian Marco; Trivisano, Carlo; Greco, Fedele Pasquale; Badiani, Anna

2017-01-01

The purpose of this study was to investigate the effect of dietary vitamin E or EconomasE™ supplementation on the growth of several background/pathogenic bacteria on rabbit carcasses and hamburgers during refrigerated storage. For 51days, 270 New Zealand rabbits received either a basal diet, or experimental diets enriched with 100 or 200mg/kg of vitamin E or EconomasE™. The bacteria studied were Salmonella, Listeria monocytogenes, Pseudomonas, Enterobacteriaceae, Escherichia coli, coagulase-positive staphylococci, plus both mesophilic and psychrotrophic aerobes. The growth of Listeria monocytogenes on contaminated patties was evaluated through a challenge test. The potential protective or antimicrobial effect of vitamin E or EconomasE™ on Listeria monocytogenes or Pseudomonas aeruginosa was assessed in vitro. Diet did not influence the concentrations of bacteria found on rabbit carcasses and developing on hamburgers. Vitamin E (in vivo and in vitro) and EconomasE™ in vivo had a protective antioxidant role, while EconomasE™ in vitro had strong antibacterial activity against Listeria monocytogenes, but not against Pseudomonas aeruginosa. Copyright © 2016 Elsevier Ltd. All rights reserved.

Challenges and Problems in the Kosovo Reality Related to Foreign Direct Investment

Directory of Open Access Journals (Sweden)

MSc. Nexhat Shkodra

2015-06-01

Full Text Available Economic development is a goal aspired by many countries of the world, Kosovo included. In attaining such goals, many countries face numerous difficulties. Amongst the most often taken paths by various countries is the attraction of foreign direct investments to the country. The term investment includes a wide range of human activities in engaging financial means into one of the areas: immoveable property, bonds and shares, manufacturing and service projects, scientific research, technological development, personnel education, etc. Different from internal investment which is engaged by domestic investors in their own territories, Foreign Direct Investment, the topic of our study, is a form of investment which generates revenues by a company in the country and an affiliate branch outside the investor’s seat. Foreign Direct Investments generate relations through the local company and its branches outside the country. Foreign Direct Investments (FDIs are considered to be a strength giving life to economic development of a country, and especially the developing countries. They have an important role to play in a long-term development of a country, and not only as a capital source, but also in increasing competitive abilities of the domestic economy, by technological transfers, strengthening infrastructure, increased productivity and generation of new employment opportunities.

CT of pulmonary emphysema - current status, challenges, and future directions

International Nuclear Information System (INIS)

Litmanovich, Diana; Boiselle, Phillip M.; Bankier, Alexander A.

2009-01-01

Pulmonary emphysema is characterized by irreversible destruction of lung parenchyma. Emphysema is a major contributor to chronic obstructive pulmonary disease (COPD), which by itself is a major cause of morbidity and mortality in the western world. Computed tomography (CT) is an established method for the in-vivo analysis of emphysema. This review first details the pathological basis of emphysema and shows how the subtypes of emphysema can be characterized by CT. The review then shows how CT is used to quantify emphysema, and describes the requirements and foundations for quantification to be accurate. Finally, the review discusses new challenges and their potential solution, notably focused on multi-detector-row CT, and emphasizes the open questions that future research on CT of pulmonary emphysema will have to address. (orig.)

Organizational Culture and Scale Development: Methodological Challenges and Future Directions

Directory of Open Access Journals (Sweden)

Bavik Ali

2014-12-01

Full Text Available Defining and measuring organizational culture (OC is of paramount importance to organizations because a strong culture could potentially increase service quality and yield sustainable competitive advantages. However, such process could be challenging to managers because the scope of OC has been defined differently across disciplines and industries, which has led to the development of various scales for measuring OC. In addition, previously developed OC scales may also not be fully applicable in the hospitality and tourism context. Therefore, by highlighting the key factors affecting the business environment and the unique characteristics of hospitality industry, this paper aims to align the scope of OC closely with the industry and to put forth the need for a new OC scale that accurately responds to the context of the hospitality industry.

The proteomic dataset for bone marrow derived human mesenchymal stromal cells: Effect of in vitro passaging

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Samuel T. Mindaye

2015-12-01

Full Text Available Bone-marrow derived mesenchymal stromal cells (BMSCs have been in clinical trials for therapy. One major bottleneck in the advancement of BMSC-based products is the challenge associated with cell isolation, characterization, and ensuring cell fitness over the course of in vitro cell propagation steps. The data in this report is part of publications that explored the proteomic changes following in vitro passaging of BMSCs [4] and the molecular heterogeneity in cultures obtained from different human donors [5,6].The methodological details involving cell manufacturing, proteome harvesting, protein identification and quantification as well as the bioinformatic analyses were described to ensure reproducibility of the results.

Scutellarin promotes in vitro angiogenesis in human umbilical vein endothelial cells

International Nuclear Information System (INIS)

Gao, Zhong-Xiu-Zi; Huang, Da-Yong; Li, Hai-Xia; Zhang, Li-Na; Lv, Yan-Hong; Cui, Hai-Dong; Zheng, Jin-Hua

2010-01-01

Research highlights: → It has been shown that scutellarin exhibits a variety of pharmacological actions, including anti-oxidative, anti-inflammatory, vasodilator as well as cardiovascular and cerebrovascular ischemia protective effects, indicating beneficial vascular effects of scutellarin. Therefore, it is speculated that scutellarin may be able to stimulate angiogenesis, which could be beneficial in the treatment of ischemic disease, wound healing and tissue regeneration. → The purpose of the present study was to elucidate the direct angiogenic actions of scutellarin on human umbilical vein endothelial cells (HUVECs) in vitro. → Our results showed that scutellarin to directly induce in vitro angiogenesis, which is closely correlated with upregulated MMP-2 expression, suggesting a potential for increasing angiogenesis. -- Abstract: Angiogenesis is critical to a wide range of physiological and pathological processes. Scutellarin, a major flavonoid of a Chinese herbal medicine Erigeron breviscapus (Vant.) Hand. Mazz. has been shown to offer beneficial effects on cardiovascular and cerebrovascular functions. However, scutellarin's effects on angiogenesis and underlying mechanisms are not fully elucidated. Here, we studied angiogenic effects of scutellarin on human umbilical vein endothelial cells (HUVECs) in vitro. Scutellarin was found by MTT assay to induce proliferation of HUVECs. In scutellarin-treated HUVECs, a dramatic increase in migration was measured by wound healing assay; Transwell chamber assay found significantly more invading cells in scutellarin-treated groups. Scutellarin also promoted capillary-like tube formation in HUVECs on Matrigel, and significantly upregulated platelet endothelial cell adhesion molecule-1 at both mRNA and protein levels. Scutellarin's angiogenic mechanism was investigated in vitro by measuring expression of angiogenic factors associated with cell migration and invasion. Scutellarin strongly induced MMP-2 activation and m

Scutellarin promotes in vitro angiogenesis in human umbilical vein endothelial cells

Energy Technology Data Exchange (ETDEWEB)

Gao, Zhong-Xiu-Zi [Department of Anatomy, Basic Medical Science College, Harbin Medical University, Harbin (China); Huang, Da-Yong [Department of Oncology, The Second Clinical Hospital, Harbin Medical University, Harbin (China); Li, Hai-Xia; Zhang, Li-Na; Lv, Yan-Hong; Cui, Hai-Dong [Department of Anatomy, Basic Medical Science College, Harbin Medical University, Harbin (China); Zheng, Jin-Hua, E-mail: jhzhenghrbmu@yahoo.cn [Department of Anatomy, Basic Medical Science College, Harbin Medical University, Harbin (China)

2010-09-10

Research highlights: {yields} It has been shown that scutellarin exhibits a variety of pharmacological actions, including anti-oxidative, anti-inflammatory, vasodilator as well as cardiovascular and cerebrovascular ischemia protective effects, indicating beneficial vascular effects of scutellarin. Therefore, it is speculated that scutellarin may be able to stimulate angiogenesis, which could be beneficial in the treatment of ischemic disease, wound healing and tissue regeneration. {yields} The purpose of the present study was to elucidate the direct angiogenic actions of scutellarin on human umbilical vein endothelial cells (HUVECs) in vitro. {yields} Our results showed that scutellarin to directly induce in vitro angiogenesis, which is closely correlated with upregulated MMP-2 expression, suggesting a potential for increasing angiogenesis. -- Abstract: Angiogenesis is critical to a wide range of physiological and pathological processes. Scutellarin, a major flavonoid of a Chinese herbal medicine Erigeron breviscapus (Vant.) Hand. Mazz. has been shown to offer beneficial effects on cardiovascular and cerebrovascular functions. However, scutellarin's effects on angiogenesis and underlying mechanisms are not fully elucidated. Here, we studied angiogenic effects of scutellarin on human umbilical vein endothelial cells (HUVECs) in vitro. Scutellarin was found by MTT assay to induce proliferation of HUVECs. In scutellarin-treated HUVECs, a dramatic increase in migration was measured by wound healing assay; Transwell chamber assay found significantly more invading cells in scutellarin-treated groups. Scutellarin also promoted capillary-like tube formation in HUVECs on Matrigel, and significantly upregulated platelet endothelial cell adhesion molecule-1 at both mRNA and protein levels. Scutellarin's angiogenic mechanism was investigated in vitro by measuring expression of angiogenic factors associated with cell migration and invasion. Scutellarin strongly

Direct Marketing Communication at SEAL Communication Company

OpenAIRE

Wachtler, Kamilla Dóra

2010-01-01

The appearance of the Internet and new, rapidly changing emerging online technologies have led to some fundamental changes in marketing -- especially direct marketing. Besides the growing popularity of the Internet, the appearance of databases and one-to-one marketing are challenging traditional marketing methods. This thesis attempts to illustrate how direct marketing techniques can be implemented in an IT company. It starts with an overview of the concepts of direct marketing and its techni...

Histamine and tryptase in nasal lavage fluid after allergen challenge

DEFF Research Database (Denmark)

Jacobi, H H; Skov, P S; Poulsen, L K

1999-01-01

BACKGROUND: Antihistamines (H1-receptor antagonists) act by competitive antagonism of histamine at H1-receptors. In addition, high concentrations of some antihistamines inhibit allergen-induced histamine release from mast cells in vitro. OBJECTIVE: The purpose of this study was to determine...... the effect of intranasal azelastine or systemic cetirizine (both potent antihistamines) on the allergen-induced release of mast-cell mediators from the human nasal mucosa in vivo. METHODS: Patients allergic to birch pollen (n = 11) and control subjects not allergic to birch pollen (n = 5) were included......, nasal allergen challenges were performed, and the number of sneezes were counted. In addition, nasal lavage fluid was collected, and the levels of mast-cell mediators (histamine and tryptase) were measured. RESULTS: The allergen challenge of patients allergic to pollen produced sneezing...

In-vitro engineering of novel bioactivity in the natural enzymes

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Vishvanath Tiwari

2016-10-01

Full Text Available Enzymes catalyze various biochemical functions with high efficiency and specificity. In-vitro design of the enzyme leads to novel bioactivity in this natural biomolecule that give answers of some vital questions like crucial residues in binding with substrate, molecular evolution, cofactor specificity etc. Enzyme engineering technology involves directed evolution, rational designing, semi-rational designing and structure-based designing using chemical modifications. Similarly, combined computational and in-vitro evolution approaches together help in artificial designing of novel bioactivity in the natural enzyme. DNA shuffling, error prone PCR and staggered extension process are used to artificially redesign active site of enzyme, which can alter its efficiency and specificity. Modifications of the enzyme can lead to the discovery of new path of molecular evolution, designing of efficient enzymes, locating active sites and crucial residues, shift in substrate and cofactor specificity. The methods and thermodynamics of in-vitro designing of the enzyme are also discussed. Similarly, engineered thermophilic and psychrophilic enzymes attain substrate specificity and activity of mesophilic enzymes that may also be beneficial for industry and therapeutics.

The use of benthic indicators in Europe: from the Water Framework Directive to the Marine Strategy Framework Directive.

Science.gov (United States)

Van Hoey, Gert; Borja, Angel; Birchenough, Silvana; Buhl-Mortensen, Lene; Degraer, Steven; Fleischer, Dirk; Kerckhof, Francis; Magni, Paolo; Muxika, Iñigo; Reiss, Henning; Schröder, Alexander; Zettler, Michael L

2010-12-01

The Water Framework Directive (WFD) and the Marine Strategy Framework Directive (MSFD) are the European umbrella regulations for water systems. It is a challenge for the scientific community to translate the principles of these directives into realistic and accurate approaches. The aim of this paper, conducted by the Benthos Ecology Working Group of ICES, is to describe how the principles have been translated, which were the challenges and best way forward. We have tackled the following principles: the ecosystem-based approach, the development of benthic indicators, the definition of 'pristine' or sustainable conditions, the detection of pressures and the development of monitoring programs. We concluded that testing and integrating the different approaches was facilitated during the WFD process, which led to further insights and improvements, which the MSFD can rely upon. Expert involvement in the entire implementation process proved to be of vital importance. Copyright © 2010 Elsevier Ltd. All rights reserved.

Optimized in vitro procedure for assessing the cytocompatibility of magnesium-based biomaterials.

Science.gov (United States)

Jung, Ole; Smeets, Ralf; Porchetta, Dario; Kopp, Alexander; Ptock, Christoph; Müller, Ute; Heiland, Max; Schwade, Max; Behr, Björn; Kröger, Nadja; Kluwe, Lan; Hanken, Henning; Hartjen, Philip

2015-09-01

Magnesium (Mg) is a promising biomaterial for degradable implant applications that has been extensively studied in vitro and in vivo in recent years. In this study, we developed a procedure that allows an optimized and uniform in vitro assessment of the cytocompatibility of Mg-based materials while respecting the standard protocol DIN EN ISO 10993-5:2009. The mouse fibroblast line L-929 was chosen as the preferred assay cell line and MEM supplemented with 10% FCS, penicillin/streptomycin and 4mM l-glutamine as the favored assay medium. The procedure consists of (1) an indirect assessment of effects of soluble Mg corrosion products in material extracts and (2) a direct assessment of the surface compatibility in terms of cell attachment and cytotoxicity originating from active corrosion processes. The indirect assessment allows the quantification of cell-proliferation (BrdU-assay), viability (XTT-assay) as well as cytotoxicity (LDH-assay) of the mouse fibroblasts incubated with material extracts. Direct assessment visualizes cells attached to the test materials by means of live-dead staining. The colorimetric assays and the visual evaluation complement each other and the combination of both provides an optimized and simple procedure for assessing the cytocompatibility of Mg-based biomaterials in vitro. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Toward the establishment of standardized in vitro tests for lipid-based formulations, part 4

DEFF Research Database (Denmark)

Williams, Hywel D; Sassene, Philip; Kleberg, Karen

2014-01-01

The Lipid Formulation Classification System Consortium looks to develop standardized in vitro tests and to generate much-needed performance criteria for lipid-based formulations (LBFs). This article highlights the value of performing a second, more stressful digestion test to identify LBFs near...... a performance threshold and to facilitate lead formulation selection in instances where several LBF prototypes perform adequately under standard digestion conditions (but where further discrimination is necessary). Stressed digestion tests can be designed based on an understanding of the factors that affect LBF...... development, and facilitate dialogue with the regulatory authorities. This classification system is based on the concept that performance evaluations across three in vitro tests, designed to subject a LBF to progressively more challenging conditions, will enable effective LBF discrimination and performance...

In vitro micropropagation of Dracaena sanderiana Sander ex Mast: An important indoor ornamental plant.

Science.gov (United States)

Aslam, Junaid; Mujib, Abdul; Sharma, Maheshwar Prasad

2013-01-01

A protocol has been developed for in vitro plant regeneration from a nodal explant of Dracaena sanderiana Sander ex Mast. Nodal explant showed high callus induction potentiality on MS medium supplemented with 6.78 μM 2,4-dichlorophenoxyacetic acid (2,4-D) followed by 46.5 μM chlorophenoxy acetic acid (CPA). The highest frequency of shoot regeneration (85%) and number of shoots per explant (5.6) were obtained on medium supplemented with 7.84 μM N(6)-benzylaminopurine (BA). Rooting was high on MS solid compared to liquid medium when added with 7.38 μM indole-3-butyric acid (IBA). Fifty percent of the roots were also directly rooted as microcuttings on soil rite, sand and peat mixture (1:1:1). In vitro and ex vitro raised plantlets were used for acclimatization. More than 90% of the plantlets was successfully acclimatized and established in plastic pots. Ex vitro transferred plantlets were normal without any phenotypic aberrations.

Hemozoin (malarial pigment directly promotes apoptosis of erythroid precursors.

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Abigail A Lamikanra

2009-12-01

Full Text Available Severe malarial anemia is the most common syndrome of severe malaria in endemic areas. The pathophysiology of chronic malaria is characterised by a striking degree of abnormal development of erythroid precursors (dyserythropoiesis and an inadequate erythropoietic response in spite of elevated levels of erythropoietin. The cause of dyserythropoiesis is unclear although it has been suggested that bone-marrow macrophages release cytokines, chemokines or lipo-peroxides after exposure to hemozoin, a crystalloid form of undigested heme moieties from malarial infected erythrocytes, and so inhibit erythropoiesis. However, we have previously shown that hemozoin may directly inhibit erythroid development in vitro and the levels of hemozoin in plasma from patients with malarial anemia and hemozoin within the bone marrow was associated with reduced reticulocyte response. We hypothesized that macrophages may reduce, not enhance, the inhibitory effect of hemozoin on erythropoiesis. In an in vitro model of erythropoiesis, we now show that inhibition of erythroid cell development by hemozoin isolated from P. falciparum is characterised by delayed expression of the erythroid markers and increased apoptosis of progenitor cells. Crucially, macrophages appear to protect erythroid cells from hemozoin, consistent with a direct contribution of hemozoin to the depression of reticulocyte output from the bone marrow in children with malarial anemia. Moreover, hemozoin isolated from P. falciparum in vitro inhibits erythroid development independently of inflammatory mediators by inducing apoptotic pathways that not only involve activation of caspase 8 and cleavage of caspase 3 but also loss of mitochondrial potential. Taken together these data are consistent with a direct effect of hemozoin in inducing apoptosis in developing erythroid cells in malarial anemia. Accumulation of hemozoin in the bone marrow could therefore result in inadequate reticulocytosis in children that

Female reproductive tract pain: targets, challenges, and outcomes

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Phillip eJobling

2014-02-01

Full Text Available Pain from the female reproductive tract (FRT is a significant clinical problem for which there are few effective therapies. The complex neuroantomy of pelvic organs not only makes diagnosis of pelvic pain disorders difficult but represents a challenge to development of targeted therapies. A number of potential therapeutic targets have been identified on sensory neurons supplying the FRT but our knowledge on the basic neurophysiology of these neurons is limited compared with other viscera. Until this is addressed we can only guess if the new experimental therapies proposed for somatic, gastrointestinal or bladder pain will translate to the FRT. Once suitable therapeutic targets become clear, the next challenge is drug delivery. The FRT represents a promising system for topical drug delivery that could be tailored to act locally or systemically depending on formulation. Development of these therapies and their delivery systems will need to be done in concert with more robust in vivo and in vitro models of FRT pain.

Marginal and internal fit of cobalt-chromium copings fabricated using the conventional and the direct metal laser sintering techniques: A comparative in vitro study.

Science.gov (United States)

Ullattuthodi, Sujana; Cherian, Kandathil Phillip; Anandkumar, R; Nambiar, M Sreedevi

2017-01-01

This in vitro study seeks to evaluate and compare the marginal and internal fit of cobalt-chromium copings fabricated using the conventional and direct metal laser sintering (DMLS) techniques. A master model of a prepared molar tooth was made using cobalt-chromium alloy. Silicone impression of the master model was made and thirty standardized working models were then produced; twenty working models for conventional lost-wax technique and ten working models for DMLS technique. A total of twenty metal copings were fabricated using two different production techniques: conventional lost-wax method and DMLS; ten samples in each group. The conventional and DMLS copings were cemented to the working models using glass ionomer cement. Marginal gap of the copings were measured at predetermined four points. The die with the cemented copings are standardized-sectioned with a heavy duty lathe. Then, each sectioned samples were analyzed for the internal gap between the die and the metal coping using a metallurgical microscope. Digital photographs were taken at ×50 magnification and analyzed using measurement software. Statistical analysis was done by unpaired t -test and analysis of variance (ANOVA). The results of this study reveal that no significant difference was present in the marginal gap of conventional and DMLS copings ( P > 0.05) by means of ANOVA. The mean values of internal gap of DMLS copings were significantly greater than that of conventional copings ( P < 0.05). Within the limitations of this in vitro study, it was concluded that the internal fit of conventional copings was superior to that of the DMLS copings. Marginal fit of the copings fabricated by two different techniques had no significant difference.

Regulation of Ghrelin Receptor by Periodontal Bacteria In Vitro and In Vivo

OpenAIRE

Nokhbehsaim, Marjan; Damanaki, Anna; Nogueira, Andressa Vilas Boas; Eick, Sigrun; Memmert, Svenja; Zhou, Xiaoyan; Nanayakkara, Shanika; Götz, Werner; Cirelli, Joni Augusto; Jäger, Andreas; Deschner, James

2017-01-01

Ghrelin plays a major role in obesity-related diseases which have been shown to be associated with periodontitis. This study sought to analyze the expression of the functional receptor for ghrelin (GHS-R1a) in periodontal cells and tissues under microbial conditions in vitro and in vivo. The GHS-R1a expression in human periodontal cells challenged with the periodontopathogen Fusobacterium nucleatum, in gingival biopsies from periodontally healthy and diseased individuals, and from rats with a...

Consumer-directed health care and the courts: let the buyer (and seller) beware.

Science.gov (United States)

Jacobson, Peter D; Tunick, Michael R

2007-01-01

In consumer-directed health care, patients will be expected to exert greater control over their spending decisions than before. As consumer-directed care gains market acceptance, courts will inevitably be involved in resolving challenges to the new arrangements. We anticipate that courts will be generally favorable toward consumer-directed care, but the new legal doctrine will not uniformly favor medical professionals and insurers. The information demands inherent in consumer-directed care will present particular legal challenges to physicians and insurers. Even as courts provide flexibility to reflect the new market realities, they will closely monitor how consumer-directed care is implemented.

Epithelial cells as active player in fibrosis: findings from an in vitro model.

Directory of Open Access Journals (Sweden)

Solange Moll

Full Text Available Kidney fibrosis, a scarring of the tubulo-interstitial space, is due to activation of interstitial myofibroblasts recruited locally or systemically with consecutive extracellular matrix deposition. Newly published clinical studies correlating acute kidney injury (AKI to chronic kidney disease (CKD challenge this pathological concept putting tubular epithelial cells into the spotlight. In this work we investigated the role of epithelial cells in fibrosis using a simple controlled in vitro system. An epithelial/mesenchymal 3D cell culture model composed of human proximal renal tubular cells and fibroblasts was challenged with toxic doses of Cisplatin, thus injuring epithelial cells. RT-PCR for classical fibrotic markers was performed on fibroblasts to assess their modulation toward an activated myofibroblast phenotype in presence or absence of that stimulus. Epithelial cell lesion triggered a phenotypical modulation of fibroblasts toward activated myofibroblasts as assessed by main fibrotic marker analysis. Uninjured 3D cell culture as well as fibroblasts alone treated with toxic stimulus in the absence of epithelial cells were used as control. Our results, with the caveats due to the limited, but highly controllable and reproducible in vitro approach, suggest that epithelial cells can control and regulate fibroblast phenotype. Therefore they emerge as relevant target cells for the development of new preventive anti-fibrotic therapeutic approaches.

In vitro cerebrovascular modeling in the 21st century: current and prospective technologies.

Science.gov (United States)

Palmiotti, Christopher A; Prasad, Shikha; Naik, Pooja; Abul, Kaisar M D; Sajja, Ravi K; Achyuta, Anilkumar H; Cucullo, Luca

2014-12-01

The blood-brain barrier (BBB) maintains the brain homeostasis and dynamically responds to events associated with systemic and/or rheological impairments (e.g., inflammation, ischemia) including the exposure to harmful xenobiotics. Thus, understanding the BBB physiology is crucial for the resolution of major central nervous system CNS) disorders challenging both health care providers and the pharmaceutical industry. These challenges include drug delivery to the brain, neurological disorders, toxicological studies, and biodefense. Studies aimed at advancing our understanding of CNS diseases and promoting the development of more effective therapeutics are primarily performed in laboratory animals. However, there are major hindering factors inherent to in vivo studies such as cost, limited throughput and translational significance to humans. These factors promoted the development of alternative in vitro strategies for studying the physiology and pathophysiology of the BBB in relation to brain disorders as well as screening tools to aid in the development of novel CNS drugs. Herein, we provide a detailed review including pros and cons of current and prospective technologies for modelling the BBB in vitro including ex situ, cell based and computational (in silico) models. A special section is dedicated to microfluidic systems including micro-BBB, BBB-on-a-chip, Neurovascular Unit-on-a-Chip and Synthetic Microvasculature Blood-brain Barrier.

An Initial Evaluation of Direct Care Staff Resilience Workshops in Intellectual Disabilities Services

Science.gov (United States)

Ingham, Barry; Riley, Jenny; Nevin, Helen; Evans, Gemma; Gair, Elodie

2013-01-01

The emotional responses to challenging behaviour of direct care staff who support people with intellectual disabilities is thought to be an important mediating factor within the stress experienced by staff and a potential maintaining factor in challenging behaviour. A brief workshop to improve direct care staff resilience was developed and…

Challenges in implementing electronic hand hygiene monitoring systems.

Science.gov (United States)

Conway, Laurie J

2016-05-02

Electronic hand hygiene (HH) monitoring systems offer the exciting prospect of a more precise, less biased measure of HH performance than direct observation. However, electronic systems are challenging to implement. Selecting a system that minimizes disruption to the physical infrastructure and to clinician workflow, and that fits with the organization's culture and budget, is challenging. Getting front-line workers' buy-in and addressing concerns about the accuracy of the system and how the data will be used are also difficult challenges. Finally, ensuring information from the system reaches front-line workers and is used by them to improve HH practice is a complex challenge. We describe these challenges in detail and suggests ways to overcome them. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Absorbable magnesium-based stent: physiological factors to consider for in vitro degradation assessments.

Science.gov (United States)

Wang, Juan; Smith, Christopher E; Sankar, Jagannathan; Yun, Yeoheung; Huang, Nan

2015-03-01

Absorbable metals have been widely tested in various in vitro settings using cells to evaluate their possible suitability as an implant material. However, there exists a gap between in vivo and in vitro test results for absorbable materials. A lot of traditional in vitro assessments for permanent materials are no longer applicable to absorbable metallic implants. A key step is to identify and test the relevant microenvironment and parameters in test systems, which should be adapted according to the specific application. New test methods are necessary to reduce the difference between in vivo and in vitro test results and provide more accurate information to better understand absorbable metallic implants. In this investigative review, we strive to summarize the latest test methods for characterizing absorbable magnesium-based stent for bioabsorption/biodegradation behavior in the mimicking vascular environments. Also, this article comprehensively discusses the direction of test standardization for absorbable stents to paint a more accurate picture of the in vivo condition around implants to determine the most important parameters and their dynamic interactions.

Development in Assay Methods for in Vitro Antimalarial Drug Efficacy Testing: A Systematic Review

Directory of Open Access Journals (Sweden)

Shweta Sinha

2017-10-01

Full Text Available The emergence and spread of drug resistance are the major challenges in malaria eradication mission. Besides various strategies laid down by World Health Organization, such as vector management, source reduction, early case detection, prompt treatment, and development of new diagnostics and vaccines, nevertheless the need for new and efficacious drugs against malaria has become a critical priority on the global malaria research agenda. At several screening stages, millions of compounds are screened (1,000–2,000,000 compounds per screening campaign, before pre-clinical trials to select optimum lead. Carrying out in vitro screening of antimalarials is very difficult as different assay methods are subject to numerous sources of variability across different laboratories around the globe. Despite this, in vitro screening is an essential part of antimalarial drug development as it enables to resource various confounding factors such as host immune response and drug–drug interaction. Therefore, in this article, we try to illustrate the basic necessity behind in vitro study and how new methods are developed and subsequently adopted for high-throughput antimalarial drug screening and its application in achieving the next level of in vitro screening based on the current approaches (such as stem cells.

Hepatocyte-based in vitro model for assessment of drug-induced cholestasis

Energy Technology Data Exchange (ETDEWEB)

Chatterjee, Sagnik, E-mail: Sagnik.Chatterjee@pharm.kuleuven.be [Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O and N2, Herestraat 49 — bus 921, 3000 Leuven (Belgium); Richert, Lysiane, E-mail: l.richert@kaly-cell.com [KaLy-Cell, 20A rue du Général Leclerc, 67115 Plobsheim (France); Augustijns, Patrick, E-mail: Patrick.Augustijns@pharm.kuleuven.be [Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O and N2, Herestraat 49 — bus 921, 3000 Leuven (Belgium); Annaert, Pieter, E-mail: Pieter.Annaert@pharm.kuleuven.be [Drug Delivery and Disposition, KU Leuven Department of Pharmaceutical and Pharmacological Sciences, O and N2, Herestraat 49 — bus 921, 3000 Leuven (Belgium)

2014-01-01

Early detection of drug-induced cholestasis remains a challenge during drug development. We have developed and validated a biorelevant sandwich-cultured hepatocytes- (SCH) based model that can identify compounds causing cholestasis by altering bile acid disposition. Human and rat SCH were exposed (24–48 h) to known cholestatic and/or hepatotoxic compounds, in the presence or in the absence of a concentrated mixture of bile acids (BAs). Urea assay was used to assess (compromised) hepatocyte functionality at the end of the incubations. The cholestatic potential of the compounds was expressed by calculating a drug-induced cholestasis index (DICI), reflecting the relative residual urea formation by hepatocytes co-incubated with BAs and test compound as compared to hepatocytes treated with test compound alone. Compounds with clinical reports of cholestasis, including cyclosporin A, troglitazone, chlorpromazine, bosentan, ticlopidine, ritonavir, and midecamycin showed enhanced toxicity in the presence of BAs (DICI ≤ 0.8) for at least one of the tested concentrations. In contrast, the in vitro toxicity of compounds causing hepatotoxicity by other mechanisms (including diclofenac, valproic acid, amiodarone and acetaminophen), remained unchanged in the presence of BAs. A safety margin (SM) for drug-induced cholestasis was calculated as the ratio of lowest in vitro concentration for which was DICI ≤ 0.8, to the reported mean peak therapeutic plasma concentration. SM values obtained in human SCH correlated well with reported % incidence of clinical drug-induced cholestasis, while no correlation was observed in rat SCH. This in vitro model enables early identification of drug candidates causing cholestasis by disturbed BA handling. - Highlights: • Novel in vitro assay to detect drug-induced cholestasis • Rat and human sandwich-cultured hepatocytes (SCH) as in vitro models • Cholestatic compounds sensitize SCH to toxic effects of accumulating bile acids • Drug

Hepatocyte-based in vitro model for assessment of drug-induced cholestasis

International Nuclear Information System (INIS)

Chatterjee, Sagnik; Richert, Lysiane; Augustijns, Patrick; Annaert, Pieter

2014-01-01

Early detection of drug-induced cholestasis remains a challenge during drug development. We have developed and validated a biorelevant sandwich-cultured hepatocytes- (SCH) based model that can identify compounds causing cholestasis by altering bile acid disposition. Human and rat SCH were exposed (24–48 h) to known cholestatic and/or hepatotoxic compounds, in the presence or in the absence of a concentrated mixture of bile acids (BAs). Urea assay was used to assess (compromised) hepatocyte functionality at the end of the incubations. The cholestatic potential of the compounds was expressed by calculating a drug-induced cholestasis index (DICI), reflecting the relative residual urea formation by hepatocytes co-incubated with BAs and test compound as compared to hepatocytes treated with test compound alone. Compounds with clinical reports of cholestasis, including cyclosporin A, troglitazone, chlorpromazine, bosentan, ticlopidine, ritonavir, and midecamycin showed enhanced toxicity in the presence of BAs (DICI ≤ 0.8) for at least one of the tested concentrations. In contrast, the in vitro toxicity of compounds causing hepatotoxicity by other mechanisms (including diclofenac, valproic acid, amiodarone and acetaminophen), remained unchanged in the presence of BAs. A safety margin (SM) for drug-induced cholestasis was calculated as the ratio of lowest in vitro concentration for which was DICI ≤ 0.8, to the reported mean peak therapeutic plasma concentration. SM values obtained in human SCH correlated well with reported % incidence of clinical drug-induced cholestasis, while no correlation was observed in rat SCH. This in vitro model enables early identification of drug candidates causing cholestasis by disturbed BA handling. - Highlights: • Novel in vitro assay to detect drug-induced cholestasis • Rat and human sandwich-cultured hepatocytes (SCH) as in vitro models • Cholestatic compounds sensitize SCH to toxic effects of accumulating bile acids • Drug

A Tox21 Approach to Altered Epigenetic Landscapes: Assessing Epigenetic Toxicity Pathways Leading to Altered Gene Expression and Oncogenic Transformation In Vitro

Directory of Open Access Journals (Sweden)

Craig L. Parfett

2017-06-01

Full Text Available An emerging vision for toxicity testing in the 21st century foresees in vitro assays assuming the leading role in testing for chemical hazards, including testing for carcinogenicity. Toxicity will be determined by monitoring key steps in functionally validated molecular pathways, using tests designed to reveal chemically-induced perturbations that lead to adverse phenotypic endpoints in cultured human cells. Risk assessments would subsequently be derived from the causal in vitro endpoints and concentration vs. effect data extrapolated to human in vivo concentrations. Much direct experimental evidence now shows that disruption of epigenetic processes by chemicals is a carcinogenic mode of action that leads to altered gene functions playing causal roles in cancer initiation and progression. In assessing chemical safety, it would therefore be advantageous to consider an emerging class of carcinogens, the epigenotoxicants, with the ability to change chromatin and/or DNA marks by direct or indirect effects on the activities of enzymes (writers, erasers/editors, remodelers and readers that convey the epigenetic information. Evidence is reviewed supporting a strategy for in vitro hazard identification of carcinogens that induce toxicity through disturbance of functional epigenetic pathways in human somatic cells, leading to inactivated tumour suppressor genes and carcinogenesis. In the context of human cell transformation models, these in vitro pathway measurements ensure high biological relevance to the apical endpoint of cancer. Four causal mechanisms participating in pathways to persistent epigenetic gene silencing were considered: covalent histone modification, nucleosome remodeling, non-coding RNA interaction and DNA methylation. Within these four interacting mechanisms, 25 epigenetic toxicity pathway components (SET1, MLL1, KDM5, G9A, SUV39H1, SETDB1, EZH2, JMJD3, CBX7, CBX8, BMI, SUZ12, HP1, MPP8, DNMT1, DNMT3A, DNMT3B, TET1, MeCP2, SETDB2, BAZ2

Foreign Direct Investments – Challenges and Perspectives for Romania

Directory of Open Access Journals (Sweden)

Cristina Batusaru

2013-05-01

Full Text Available In the context of globalization of markets, foreign direct investments have an important role in terms of supporting endogenous growth factors, on the one hand and the circuit of financial flows between countries, on the other hand. If we refer to the effects of the economic crisis on economies, ISD may represent capital infusion instruments for affected economic sectors, contributing to faster recovery of economic gaps that occurred. By studying this topic of FDI we consider the great impact and benefits that they can bring, being essential element in the development of a country, as in the case of Romania. This paper presents the main trends of international financial flows for the period 2008-2012, whereas in the context of economic globalization requires an overall analysis of country-specific FDI performance as it helps in improving and optimizing strategies adopted by foreign transnational companies. In order to underline their importance and necessity, we study the situation of Romania in this field by analyzing the performance of countries in attracting direct foreign investments. Following the study conducted it has been made a number of conclusions and recommendations on how to improve this process in Romania. Academics, researchers, administrators of the university all have a great responsibility on how they support to attract FDI in Romania, even if we refer to work force that they form, the ideas they can provide in supporting and developing this process or by sharing the “know-how” related to the many fields that FDI can have an impact on. This paper aims to bring on the loop the main strengths and weaknesses that Romania has in the field of FDS and invites the readers interested on the topic to involve by providing feedback in order to improve this process in Romania.

The revision of the European blood directives: A major challenge for transfusion medicine.

Science.gov (United States)

Folléa, G; Aranko, K

2015-08-01

Using both patient-focused and donor-focused perspectives, to review the current EU blood directives, in order to derive proposals, in principle, for what should evolve during the revision process of these directives. Review of the EU blood directives in the light of scientific literature, related reports from the Directorate General Health and Consumers (DG SANTÉ), and from the Council of Europe (CoE). The analyses led us to present the main following proposals: developing voluntary unpaid donations: the directives should consider taking into consideration ethically acceptable forms of compensation consistent with altruistic donation (including plasma donations for fractionation); current expertise: more extensive utilization of the expertise of blood establishments and their consultants should be considered; donor selection: an evidence-based approach for basing donor deferral criteria on sound scientific evidence should be promoted; donor reactions: measures to prevent donor reactions and to make donations safer for the donors should also be included; quality control: The quality control requirements should relate to the Council of Europe Blood Guide specifications: these should become minimum standards (as is the case with monographs of the European Pharmacopeia), facilitating regular update of blood component lists and related specifications and compliance with the specifications; haemovigilance: because of reporting difficulties (e.g. lack of number of blood products transfused), the effectiveness of haemovigilance has so far been limited. This should lead appropriate bodies to investigate alternative or complementary ways to help improve patient safety, taking into consideration, in principle, patient blood management and the appropriate use of blood products. Furthermore, donor vigilance, which is still absent from the current directive should be included in a revised directive. These proposals for revising the current EU blood directives (if taken into

Looking at the end-of-life directive and challenges in recycling

DEFF Research Database (Denmark)

Smink, Carla

2004-01-01

The article deals with end-of-life vechile regulations. Focus is on the European end-of-life vechile directive (2000/53/EC) and how Denmark and the Netherlands have dealt with the regulations in the past 30 years....

Current ethical and legal issues in health-related direct-to-consumer genetic testing.

Science.gov (United States)

Niemiec, Emilia; Kalokairinou, Louiza; Howard, Heidi Carmen

2017-09-01

A variety of health-related genetic testing is currently advertized directly to consumers. This article provides a timely overview of direct-to-consumer genetic testing (DTC GT) and salient ethical issues, as well as an analysis of the impact of the recently adopted regulation on in vitro diagnostic medical devices on DTC GT. DTC GT companies currently employ new testing approaches, report on a wide spectrum of conditions and target new groups of consumers. Such activities raise ethical issues including the questionable analytic and clinical validity of tests, the adequacy of informed consent, potentially misleading advertizing, testing in children, research uses and commercialization of genomic data. The recently adopted regulation on in vitro diagnostic medical devices may limit the offers of predisposition DTC GT in the EU market.

Challenges in effect-directed analysis with a focus on biological samples

NARCIS (Netherlands)

Simon, E.; Lamoree, M.H.; Hamers, T.; de Boer, J.

2015-01-01

Effect-directed analysis (EDA), the combined use of bioassay-guided fractionation and analytical chemical techniques, enables detection of chemicals by their effects, facilitates identification of non-target compounds and transformation products with a certain toxic mode of action and assists in

A Unifying Approach to Goal-Directed Evaluation

DEFF Research Database (Denmark)

Danvy, Olivier; Grobauer, Bernd; Rhiger, Morten

2001-01-01

semantics coincides with Gudeman’s continuation semantics of Icon. We then compile Icon programs by specializing their interpreter (i.e., by using the first Futamura projection), using type-directed partial evaluation. Through various back ends, including a run-time code generator, we generate ML code, C......Goal-directed evaluation, as embodied in Icon and Snobol, is built on the notions of backtracking and of generating successive results, and therefore it has always been something of a challenge to specify and implement. In this article, we address this challenge using computational monads...... code, and OCaml byte code. Binding-time analysis and partial evaluation of the continuation-based interpreter automatically give rise to C programs that coincide with the result of Proebsting’s optimized compiler....

Nano- and microstructured materials for in vitro studies of the physiology of vascular cells

OpenAIRE

Alexandra M. Greiner; Adria Sales; Hao Chen; Sarah A. Biela; Dieter Kaufmann; Ralf Kemkemer

2016-01-01

The extracellular environment of vascular cells in vivo is complex in its chemical composition, physical properties, and architecture. Consequently, it has been a great challenge to study vascular cell responses in vitro, either to understand their interaction with their native environment or to investigate their interaction with artificial structures such as implant surfaces. New procedures and techniques from materials science to fabricate bio-scaffolds and surfaces have enabled novel studi...

Leadership challenges in multinational medical peacekeeping operations: Lessons from UNIFIL Hospital.

Science.gov (United States)

Datta, Rakesh; Khanna, Sangeeta

2017-10-01

Commanding a military multinational and multilingual healthcare facility can be a formidable task with very little margin for error. The authors were in leadership positions of UNIFIL Hospital, unique in its diversity of both staff and clientele. Experience about the challenges faced and methods adopted to overcome them will be shared. Troops from diverse backgrounds differ in their competency, and also in their attitudinal approach to situations. It is imperative for the medical commanders to identify these differences, and work towards harnessing individual strengths to form a cohesive unit. Frequent rotation of team members and thereby difficulty in adapting to new environment makes the tasks more challenging. Challenges can be broadly categorized in those dealing with functional roles (providing medical support) and command and control issues. Linguistic challenges especially in situations where professionals have to work as a coordinated unit remains a major challenge. The threat of medical errors arising out of misunderstandings is very real. Gender sensitization is essential to avoid potential unpleasant situations. Interpersonal conflict can easily go out of hand. The leadership has to be more direct and deliberate relying less on hierarchy and more on direct communication. A strict enforcement of UN standards for equipment and competence, frequent joint medical drills help to overcome interoperability issues and develop mutual confidence. Leadership in multinational UN hospitals is a demanding task with its peculiar set of challenges. A systematic and deliberate approach focused on mutual respect, flexibility and direct leadership can help medical commanders in such situations.

Directed Differentiation of Human Embryonic Stem Cells into Prostate Organoids In Vitro and its Perturbation by Low-Dose Bisphenol A Exposure.

Directory of Open Access Journals (Sweden)

Esther L Calderon-Gierszal

Full Text Available Studies using rodent and adult human prostate stem-progenitor cell models suggest that developmental exposure to the endocrine disruptor Bisphenol-A (BPA can predispose to prostate carcinogenesis with aging. Unknown at present is whether the embryonic human prostate is equally susceptible to BPA during its natural developmental window. To address this unmet need, we herein report the construction of a pioneer in vitro human prostate developmental model to study the effects of BPA. The directed differentiation of human embryonic stem cells (hESC into prostatic organoids in a spatial system was accomplished with precise temporal control of growth factors and steroids. Activin-induced definitive endoderm was driven to prostate specification by combined exposure to WNT10B and FGF10. Matrigel culture for 20-30 days in medium containing R-Spondin-1, Noggin, EGF, retinoic acid and testosterone was sufficient for mature prostate organoid development. Immunofluorescence and gene expression analysis confirmed that organoids exhibited cytodifferentiation and functional properties of the human prostate. Exposure to 1 nM or 10 nM BPA throughout differentiation culture disturbed early morphogenesis in a dose-dependent manner with 1 nM BPA increasing and 10 nM BPA reducing the number of branched structures formed. While differentiation of branched structures to mature organoids seemed largely unaffected by BPA exposure, the stem-like cell population increased, appearing as focal stem cell nests that have not properly entered lineage commitment rather than the rare isolated stem cells found in normally differentiated structures. These findings provide the first direct evidence that low-dose BPA exposure targets hESC and perturbs morphogenesis as the embryonic cells differentiate towards human prostate organoids, suggesting that the developing human prostate may be susceptible to disruption by in utero BPA exposures.

Immunomodulatory Effects of Domoic Acid Differ Between In vivo and In vitro Exposure in Mice

Directory of Open Access Journals (Sweden)

Milton Levin

2008-12-01

Full Text Available The immunotoxic potential of domoic acid (DA, a well-characterized neurotoxin, has not been fully investigated. Phagocytosis and lymphocyte proliferation were evaluated following in vitro and in vivo exposure to assay direct vs indirect effects. Mice were injected intraperitoneally with a single dose of DA (2.5 µg/g b.w. and sampled after 12, 24, or 48 hr. In a separate experiment, leukocytes and splenocytes were exposed in vitro to 0, 1, 10, or 100 µM DA. In vivo exposure resulted in a significant increase in monocyte phagocytosis (12-hr, a significant decrease in neutrophil phagocytosis (24-hr, a significant decrease in monocyte phagocytosis (48-hr, and a significant reduction in T-cell mitogen-induced lymphocyte proliferation (24-hr. In vitro exposure significantly reduced neutrophil and monocyte phagocytosis at 1 µM. B- and T-cell mitogen-induced lymphocyte proliferation were both significantly increased at 1 and 10 µM, and significantly decreased at 100 µM. Differences between in vitro and in vivo results suggest that DA may exert its immunotoxic effects both directly and indirectly. Modulation of cytosolic calcium suggests that DA exerts its effects through ionotropic glutamate subtype surface receptors at least on monocytes. This study is the first to identify DA as an immunotoxic chemical in a mammalian species.

Response to challenging dose of x-rays as a predictive assay for molecular epidemiology

International Nuclear Information System (INIS)

Cebulska-Wasilewska, A.

2003-01-01

Human biomonitoring, as a tool to identify or quantify the potential risk from genotoxic exposures, has gained increasing interest especially in the areas of cancer risk assessment and diseases treatment. Chromosome aberrations resulting from direct DNA breakage or from inhibition of DNA repair or synthesis, measured in peripheral blood lymphocytes have been used in occupational health surveillance programs in order to assess risks from exposures. Many results in our human monitoring studies have shown influence of the environmental or occupational exposure on the cytogenetic damage detected in lymphocytes, confirming both, the association with adverse health outcome and the influence of life style related confounding factors. Susceptibility to the environmental agent actions was also evaluated in lymphocytes in the studies of variation between responses to the challenging dose of UV or X-rays followed by the evaluation of the repair capacity of the DNA damage induced by a challenging dose. The induced and residual DNA damage was analyzed with the use of SCGE assay. Susceptibility and repair capacities of healthy donors and cancer patients were compared. Studies have shown a good correlation between DNA damage induced in vivo or in vitro and cytogenetic measures. Results from studies on susceptibilities and repair competence performed in occupationally exposed and unexposed 475 healthy donors and patients with diagnosed cancer are discussed. Significantly lower efficiency of repair process was observed in cancer patients. The possible effects on repair competency of various occupational exposures and influence of the diet and other confounding factors is shown. Although in our preliminary studies comet assay failed to detect DNA damage repair disorders in a teratoma immature infant, though, prospective use of a challenging dose of radiation combined with the comet assay as a predictive assay is suggested and limitation discussed

Failure to protect calves against Taenia saginata using antigens prepared from in vitro cultivation of the larval stage.

Science.gov (United States)

Mitchell, G B; Armour, J

1980-11-01

Calves were vaccinated intramuscularly against the tapeworm Taenia saginata using excretory/secretory (ES) antigens from short and long term periods of in vitro cultivation of the larval stage of the parasite, four weeks before challenge with 5000 T saginata onchospheres. Neither immunisation regime employed afforded significant protection against challenge. It was considered that this may have been due to a reduction in concentration of, or detrimental effects to, potential immunogens during vaccine production. Elucidation of the nature of the protective ES antigens necessary for standardization of the technique has yet to be achieved in helminths.

Directional Total Generalized Variation Regularization for Impulse Noise Removal

DEFF Research Database (Denmark)

Kongskov, Rasmus Dalgas; Dong, Yiqiu

2017-01-01

this regularizer for directional images is highly advantageous. In order to estimate directions in impulse noise corrupted images, which is much more challenging compared to Gaussian noise corrupted images, we introduce a new Fourier transform-based method. Numerical experiments show that this method is more...

Direct Differentiation of Human Pluripotent Stem Cells into Haploid Spermatogenic Cells

Directory of Open Access Journals (Sweden)

Charles A. Easley, IV

2012-09-01

Full Text Available Human embryonic stem cells (hESCs and induced pluripotent stem cells (hiPSCs have been shown to differentiate into primordial germ cells (PGCs but not into spermatogonia, haploid spermatocytes, or spermatids. Here, we show that hESCs and hiPSCs differentiate directly into advanced male germ cell lineages, including postmeiotic, spermatid-like cells, in vitro without genetic manipulation. Furthermore, our procedure mirrors spermatogenesis in vivo by differentiating PSCs into UTF1-, PLZF-, and CDH1-positive spermatogonia-like cells; HIWI- and HILI-positive spermatocyte-like cells; and haploid cells expressing acrosin, transition protein 1, and protamine 1 (proteins that are uniquely found in spermatids and/or sperm. These spermatids show uniparental genomic imprints similar to those of human sperm on two loci: H19 and IGF2. These results demonstrate that male PSCs have the ability to differentiate directly into advanced germ cell lineages and may represent a novel strategy for studying spermatogenesis in vitro.

Construction of a cDNA library from human retinal pigment epithelial cells challenged with rod outer segments.

Science.gov (United States)

Cavaney, D M; Rakoczy, P E; Constable, I J

1995-05-01

To study genes expressed by retinal pigment epithelial (RPE) cells during phagocytosis and digestion of rod outer segments (ROS), a complementary (c)DNA library was produced using an in-vitro model. The cDNA library can be used to study molecular changes which contribute to the development of diseases due to a failure in outer segment phagocytosis and digestion by RPE cells. Here we demonstrate a way to study genes and their functions using a molecular biological approach and describing the first step involved in this process, the construction of a cDNA library. Human RPE cells obtained from the eyes of a seven-year-old donor were cultured and challenged with bovine ROS. The culture was harvested and total RNA was extracted. Complementary DNA was transcribed from the messenger (m)RNA and was directionally cloned into the LambdaGEM-4 bacteriophage vector successfully. Some clones were picked and the DNA extracted, to determine the size of the inserts as a measure of the quality of the library. Molecular biology and cell culture are important tools to be used in eye research, especially in areas where tissue is limiting and animal models are not available. We now have a ROS challenged RPE cDNA library which will be used to identify genes responsible for degrading phagocytosed debris within the retinal pigment epithelium.

In vitro evaluation of nutrients that selectively confer a competitive advantage to lactobacilli.

Science.gov (United States)

Vongsa, R A; Minerath, R A; Busch, M A; Tan, J; Koenig, D W

2016-01-01

An assay was developed that tested the ability of Lactobacillus acidophilus to outcompete a challenge of Escherichia coli in a mixed culture containing different test nutrients. Using this assay, addition of fructo-oligosaccharide to the media allowed L. acidophilus to outcompete a challenge of E. coli, whereas in a mixed culture without the prebiotic the trend was reversed. Growth curves generated for E. coli in a single culture showed that fructo-oligosaccharide did not affect growth, indicating that the carbohydrate was not toxic to E. coli. This indicates that fructo-oligosaccharides may increase the ability of beneficial microbes to outcompete a pathogenic challenge. These results were confirmed using a skin simulant model that incorporates growth of the organisms at an air-surface interface to mimic the vulvar environment. It is possible to use a co-culture assay as an in vitro screening tool to define nutrients that confer a competitive advantage to beneficial flora specific to the female urogenital tract.

Neutrophil-induced human bronchial hyperresponsiveness in vitro--pharmacological modulation.

Science.gov (United States)

Hughes, J M; McKay, K O; Johnson, P R; Tragoulias, S; Black, J L; Armour, C L

1993-04-01

Although it has been postulated that inflammatory cells cause the bronchial hyperresponsiveness which is diagnostic of asthma, until recently there has been little direct evidence of such a link. We have recently shown that calcium ionophore-activated human neutrophils and eosinophils can induce a state of human airway hyperresponsiveness in vitro. In this study we have shown that the anti-inflammatory agent nedocromil sodium, 10(-7) M, inhibited the hyperresponsiveness induced by products released from ionophore activated neutrophils but did not inhibit the release of leukotriene B4 from the same cells. Neutrophil-induced bronchial hyperresponsiveness was also inhibited by pre-treatment of the bronchial tissues with a thromboxane A2 and prostaglandin receptor antagonist, GR32191, 10(-7) M. These findings indicate that cyclooxygenase products are involved in bronchial hyperresponsiveness induced by inflammatory cell products in vitro and that their release can be inhibited by nedocromil sodium.

[Synergistic effect of cell kinetics-directed chemo-endocrine therapy on experimental mammary tumors].

Science.gov (United States)

Ueki, H

1987-11-01

We tried to demonstrate that the cell kinetics-directed chemoendocrine therapy is more effective on hormone dependent breast cancer than empirical combination of the endocrine therapy and chemotherapy. Cell kinetics of each tumor was measured by flow cytometric analysis. Estrogen dependent human breast cancer cell line MCF-7 was used in vitro. In vivo, androgen dependent SC-115 carcinoma was transplanted to DDS mice. In vitro, tamoxifen was administered as the endocrine therapy. In vivo, we carried out testectomy on DDS mice. Effect of the endocrine therapy on the cell kinetics of the tumor was thought to be G1-S depression. High density 5FU was administered as the chemotherapeutic agents, whose content was 1 microgram/ml in vitro and 40 mg/kg in vivo. 5FU brought temporary decrease of cells in S phase. Only anteceding 5FU administration had synergistic effect in combination of 5FU and the endocrine therapy. 5FU was convinced to act more effectively on cells in S phase, so it was shown that cell kinetics-directed schedule was superior to the empirical treatment schedule in chemoendocrine therapy.

Effects of sex steroids, sex, and sexual maturity on cortisol production: an in vitro comparison of chinook salmon and rainbow trout interrenals.

Science.gov (United States)

McQuillan, H James; Lokman, P Mark; Young, Graham

2003-08-01

Sex steroids appear to be responsible for hyperactivation of the hypothalamus-pituitary-interrenal (HPI) axis that occurs in mature semelparous Pacific salmon as a prelude to post-spawning (programmed) death. This study was undertaken to examine the direct effects of sex steroids on interrenal activity of semelparous (chinook salmon) and iteroparous (rainbow trout) salmonids using an in vitro incubation system. In addition, phenotypic sex differences in cortisol production by interrenals of sexually mature (spawning) rainbow trout and chinook salmon were investigated. Interrenal tissue from juvenile and sexually mature chinook salmon and rainbow trout was incubated for 48 h in culture medium containing either no steroid (controls), 1 microM estradiol (E2) or 1 microM 11-ketotestosterone (11-KT). This tissue was then challenged for 3h with either pregnenolone, dibutyryladenosine 3('):5(')-cyclic monophosphate (dbcAMP) or forskolin, or synthetic human adrenocorticotropic hormone (ACTH(1-24)). Sex differences in in vitro interrenal cortisol production were assessed using separate tissue pools challenged with the same agents. Cortisol in media was measured by radioimmunoassay. E2 suppressed the ability of juvenile chinook salmon interrenals to utilize pregnenolone as substrate for cortisol synthesis. In mature female chinook salmon the suppressive effect of E2 was less pronounced, but was observed as a reduced response of interrenals to both pregnenolone and dbcAMP. E2 did not affect ACTH(1-24) stimulated cortisol production. Immature and mature rainbow trout interrenals were both relatively insensitive to E2. 11-KT did not affect cortisol production by juvenile chinook salmon and juvenile or mature rainbow trout, and had only minor effects in male and female spawning chinook salmon. In mature chinook salmon and rainbow trout, the interrenals of females were more responsive to ACTH stimulation and showed a greater utilization of pregnenolone as a substrate than

Integrated deterministic and probabilistic safety assessment: Concepts, challenges, research directions

International Nuclear Information System (INIS)

Zio, Enrico

2014-01-01

Highlights: • IDPSA contributes to robust risk-informed decision making in nuclear safety. • IDPSA considers time-dependent interactions among component failures and system process. • Also, IDPSA considers time-dependent interactions among control and operator actions. • Computational efficiency by advanced Monte Carlo and meta-modelling simulations. • Efficient post-processing of IDPSA output by clustering and data mining. - Abstract: Integrated deterministic and probabilistic safety assessment (IDPSA) is conceived as a way to analyze the evolution of accident scenarios in complex dynamic systems, like nuclear, aerospace and process ones, accounting for the mutual interactions between the failure and recovery of system components, the evolving physical processes, the control and operator actions, the software and firmware. In spite of the potential offered by IDPSA, several challenges need to be effectively addressed for its development and practical deployment. In this paper, we give an overview of these and discuss the related implications in terms of research perspectives

Integrated deterministic and probabilistic safety assessment: Concepts, challenges, research directions

Energy Technology Data Exchange (ETDEWEB)

Zio, Enrico, E-mail: enrico.zio@ecp.fr [Ecole Centrale Paris and Supelec, Chair on System Science and the Energetic Challenge, European Foundation for New Energy – Electricite de France (EDF), Grande Voie des Vignes, 92295 Chatenay-Malabry Cedex (France); Dipartimento di Energia, Politecnico di Milano, Via Ponzio 34/3, 20133 Milano (Italy)

2014-12-15

Highlights: • IDPSA contributes to robust risk-informed decision making in nuclear safety. • IDPSA considers time-dependent interactions among component failures and system process. • Also, IDPSA considers time-dependent interactions among control and operator actions. • Computational efficiency by advanced Monte Carlo and meta-modelling simulations. • Efficient post-processing of IDPSA output by clustering and data mining. - Abstract: Integrated deterministic and probabilistic safety assessment (IDPSA) is conceived as a way to analyze the evolution of accident scenarios in complex dynamic systems, like nuclear, aerospace and process ones, accounting for the mutual interactions between the failure and recovery of system components, the evolving physical processes, the control and operator actions, the software and firmware. In spite of the potential offered by IDPSA, several challenges need to be effectively addressed for its development and practical deployment. In this paper, we give an overview of these and discuss the related implications in terms of research perspectives.

Optimization of jenipapo in vitro seed germination process

Directory of Open Access Journals (Sweden)

Rafaela Ribeiro de Souza

Full Text Available ABSTRACT The in vitro seed germination is an effective alternative for quickly obtaining explants with sanitary quality. However, jenipapo seeds present slow and uneven germination. Therefore, internal and external factors to seed which directly interfere in the process, they must be identified, in order to adapt better techniques to obtain seedlings. In this sense, this work aimed to optimize the in vitro germination of Genipa americana L. seeds by evaluating different factors (light quality, GA3 treatment, pre-soaking in distilled water, growing media and stratification in the dark. It was found that the seed germination of G. americana was indifferent to light, however, the best results were obtained under conditions of continuous darkness; There was no effect of the application of exogenous GA3; The pre-soaking in distilled water for 48 h contributes to obtaining better germination rates; And the reduction in MS medium salts, and laminating the pretreatment in the dark maximizes the germination potential of seeds.Therefore, the optimal conditions for in vitro germination of G. americana L. seeds requires pre-soaking in distilled water for 48 hours and inoculation into culture media consisting of 1/2 MS + 15 g L-1 sucrose, with stratification in the dark for 16 days, followed by the transfer to growth chambers with lighting provided by white fluorescent lamps.

Presidential Green Chemistry Challenge: 2007 Greener Reaction Conditions Award

Science.gov (United States)

Presidential Green Chemistry Challenge 2007 award winner, Headwaters Technology Innovation, developed a metal nanocatalyst to synthesize hydrogen peroxide directly from hydrogen and oxygen, eliminating hazardous chemicals.

Challenges and Opportunities in Teaching Mathematics

Science.gov (United States)

Goos, Merrilyn

2016-01-01

At the special conference celebrating the 50th anniversary of the Australian Association of Mathematics Teachers (AAMT), the author was asked to speak about the challenges and opportunities in teaching mathematics as a stimulus for discussion of AAMT's "future directions". With an open invitation to be a little provocative, the author…

Impact of host cell variation on the neutralization of HIV-1 in vitro.

Science.gov (United States)

Polonis, Victoria R; Schuitemaker, Hanneke; Bunnik, Evelien M; Brown, Bruce K; Scarlatti, Gabriella

2009-09-01

In this review we present current advances in our understanding of HIV-1 neutralization assays that employ primary cell types, as compared with those that utilize cell lines and the newer, more standardized pseudovirus assays. A commentary on the challenges of standardizing in-vitro neutralization assays using primary cells is included. The data from reporter cell line neutralization assays may agree with results observed in primary cells; however, exceptions have recently been reported. Multiple variables exist in primary cell assays using peripheral blood mononuclear cells from HIV-seronegative donors; in-vitro neutralization titers can vary significantly based on the donor cells used for assay targets and for virus propagation. Thus, more research is required to achieve validated primary cell neutralization assays. HIV-vaccine-induced antibody performance in the current neutralization assays may function as a 'gatekeeper' for HIV-1 subunit vaccine advancement. Development of standardized platforms for reproducible measurement of in-vitro neutralization is therefore a high priority. Given the considerable variation in results obtained from some widely applied HIV neutralization platforms, parallel evaluation of new antibodies using different host cells for assay targets, as well as virus propagation, is recommended until immune correlates of protection are identified.

A DNA-based pattern classifier with in vitro learning and associative recall for genomic characterization and biosensing without explicit sequence knowledge.

Science.gov (United States)

Lee, Ju Seok; Chen, Junghuei; Deaton, Russell; Kim, Jin-Woo

2014-01-01

Genetic material extracted from in situ microbial communities has high promise as an indicator of biological system status. However, the challenge is to access genomic information from all organisms at the population or community scale to monitor the biosystem's state. Hence, there is a need for a better diagnostic tool that provides a holistic view of a biosystem's genomic status. Here, we introduce an in vitro methodology for genomic pattern classification of biological samples that taps large amounts of genetic information from all genes present and uses that information to detect changes in genomic patterns and classify them. We developed a biosensing protocol, termed Biological Memory, that has in vitro computational capabilities to "learn" and "store" genomic sequence information directly from genomic samples without knowledge of their explicit sequences, and that discovers differences in vitro between previously unknown inputs and learned memory molecules. The Memory protocol was designed and optimized based upon (1) common in vitro recombinant DNA operations using 20-base random probes, including polymerization, nuclease digestion, and magnetic bead separation, to capture a snapshot of the genomic state of a biological sample as a DNA memory and (2) the thermal stability of DNA duplexes between new input and the memory to detect similarities and differences. For efficient read out, a microarray was used as an output method. When the microarray-based Memory protocol was implemented to test its capability and sensitivity using genomic DNA from two model bacterial strains, i.e., Escherichia coli K12 and Bacillus subtilis, results indicate that the Memory protocol can "learn" input DNA, "recall" similar DNA, differentiate between dissimilar DNA, and detect relatively small concentration differences in samples. This study demonstrated not only the in vitro information processing capabilities of DNA, but also its promise as a genomic pattern classifier that could

Urban growth and water access in sub-Saharan Africa: Progress, challenges, and emerging research directions.

Science.gov (United States)

Dos Santos, S; Adams, E A; Neville, G; Wada, Y; de Sherbinin, A; Mullin Bernhardt, E; Adamo, S B

2017-12-31

For the next decade, the global water crisis remains the risk of highest concern, and ranks ahead of climate change, extreme weather events, food crises and social instability. Across the globe, nearly one in ten people is without access to an improved drinking water source. Least Developed Countries (LDCs) especially in sub-Saharan Africa (SSA) are the most affected, having disproportionately more of the global population without access to clean water than other major regions. Population growth, changing lifestyles, increasing pollution and accelerating urbanization will continue to widen the gap between the demand for water and available supply especially in urban areas, and disproportionately affect informal settlements, where the majority of SSA's urban population resides. Distribution and allocation of water will be affected by climate-induced water stresses, poor institutions, ineffective governance, and weak political will to address scarcity and mediate uncertainties in future supply. While attempts have been made by many scientists to examine different dimensions of water scarcity and urban population dynamics, there are few comprehensive reviews, especially focused on the particular situation in Sub-Saharan Africa. This paper contributes to interdisciplinary understanding of urban water supply by distilling and integrating relevant empirical knowledge on urban dynamics and water issues in SSA, focusing on progress made and associated challenges. It then points out future research directions including the need to understand how alternatives to centralized water policies may help deliver sustainable water supply to cities and informal settlements in the region. Copyright © 2017 Elsevier B.V. All rights reserved.

Direct monitoring of dopamine and 5-HT release in substantia nigra and ventral tegmental area in vitro

DEFF Research Database (Denmark)

Rice, M E; Richards, C D; Nedergaard, S

1994-01-01

Fast-scan cyclic voltammetry with carbon fibre microelectrodes was used to detect endogenous dopamine (DA) and 5-hydroxytryptamine (5-HT) release from three distinct regions of guinea-pig mid-brain in vitro: rostral and caudal substantia nigra (SN) and the ventral tegmental area (VTA). Previous...... these regions with in situ electrodes and demonstrates the utility of fast-scan cyclic voltammetry to investigate the mechanisms and possible non-classical functions of somato-dendritic DA release....

Family-Directed Transition Planning Guide.

Science.gov (United States)

Colorado State Dept. of Education, Denver. Div. of Special Education Services.

This guide to family-directed transition planning is intended to help parents and students with disabilities take leading roles in the process of transition from school to post-school activities. First, a letter to families examines the challenge of change and the transition process. Section 2 examines regulations that affect transition planning,…

Advanced multiphoton methods for in vitro and in vivo functional imaging of mouse retinal neurons (Conference Presentation)

Science.gov (United States)

Cohen, Noam; Schejter, Adi; Farah, Nairouz; Shoham, Shy

2016-03-01

Studying the responses of retinal ganglion cell (RGC) populations has major significance in vision research. Multiphoton imaging of optogenetic probes has recently become the leading approach for visualizing neural populations and has specific advantages for imaging retinal activity during visual stimulation, because it leads to reduced direct photoreceptor excitation. However, multiphoton retinal activity imaging is not straightforward: point-by-point scanning leads to repeated neural excitation while optical access through the rodent eye in vivo has proven highly challenging. Here, we present two enabling optical designs for multiphoton imaging of responses to visual stimuli in mouse retinas expressing calcium indicators. First, we present an imaging solution based on Scanning Line Temporal Focusing (SLITE) for rapidly imaging neuronal activity in vitro. In this design, we scan a temporally focused line rather than a point, increasing the scan speed and reducing the impact of repeated excitation, while maintaining high optical sectioning. Second, we present the first in vivo demonstration of two-photon imaging of RGC activity in the mouse retina. To obtain these cellular resolution recordings we integrated an illumination path into a correction-free imaging system designed using an optical model of the mouse eye. This system can image at multiple depths using an electronically tunable lens integrated into its optical path. The new optical designs presented here overcome a number of outstanding obstacles, allowing the study of rapid calcium- and potentially even voltage-indicator signals both in vitro and in vivo, thereby bringing us a step closer toward distributed monitoring of action potentials.

Presidential Green Chemistry Challenge: 2000 Small Business Award

Science.gov (United States)

Presidential Green Chemistry Challenge 2000 award winner, RevTech, developed a process to print top-quality labels directly on glass. Their Envirogluv inks have no heavy metals, have little to no VOCs, and are biodegradable.

Ligand-directed tosyl chemistry for in situ native protein labeling and engineering in living systems: from basic properties to applications.

Science.gov (United States)

Tsukiji, Shinya; Hamachi, Itaru

2014-08-01

The ability to introduce any chemical probe to any endogenous target protein in its native environment, that is in cells and in vivo, is anticipated to provide various new exciting tools for biological and biomedical research. Although still at the prototype stage, the ligand-directed tosyl (LDT) chemistry is a novel type of affinity labeling technique that we developed for such a dream. This chemistry allows for modifying native proteins by various chemical probes with high specificity in various biological settings ranging from in vitro (in test tubes) to in living cells and in vivo. Since the first report, the list of proteins that are successfully labeled by the LDT chemistry has been increasing. A growing number of studies have demonstrated its utility to create semisynthetic proteins directly in cellular contexts. The in situ generated semisynthetic proteins are applicable for various types of analysis and imaging of intracellular biological processes. In this review, we summarize the basic properties of the LDT chemistry and its applications toward in situ engineering and analysis of native proteins in living systems. Current limitations and future challenges of this area are also described. Copyright © 2014 Elsevier Ltd. All rights reserved.

Antiviral activities of Radix Isatidis polysaccharide against type II herpes simplex virus in vitro

Directory of Open Access Journals (Sweden)

Chunmei WANG

2018-03-01

Full Text Available Abstract This study investigated the antiviral activities of Radix Isatidis polysaccharide (RIP against type II herpes simplex virus (HSV-2 in vitro. RIP was prepared from the Radix Isatidis root. The toxicity of RIP on Vero cells was detected. The direct killing effect of RIP on HSV-2, inhibitory effect of RIP on HSV-2 replication and inhibitory effect of RIP on HSV-2 adsorption were determined. Results showed that, RIP in concentration range of 25-800 mg/L had no toxic effect on Vero cells. RIP with different concentrations could not directly inactivate the HSV-2. The effective rates on inhibition of HSV-2 replication and adsorption in 800 mg/L RIP group were 71.57% and 48.37%, respectively, which were the highest among different groups. In conclusion, RIP has the antiviral effect against HSV-2 in vitro. This effect mainly occurs in inhibiting the virus duplication and adsorption.

Challenges in Request Management

DEFF Research Database (Denmark)

Sommer, Anita Friis

2014-01-01

and its customers. The study provides an insight into a new area of supply chain management, including the process activity flow and challenges involved across the process. Furthermore, the method is dyadic including the customer in the case study, which is rare in related research....... profitability. This research study seeks to investigate the challenges of RQM in practice. Existing demand chain management literature is used as a basis for developing a RQM framework. RQM is investigated through an explorative research design in a dyadic B2B case study including a global industrial company......Request management (RQM) is a new term used for managing customer requests for new products. It is the counterpart to typical product development processes, which has no direct customer involvement. It is essential to manage customer requests in a structured and efficient way to obtain...

Study in vitro of origin radioprotective food the radioprotective effect in vitro of food borne; Estudio in vitro de radioprotectores de origen alimentario

Energy Technology Data Exchange (ETDEWEB)

Soraino, J. M.; Sebastia, N.; Almonacid, M.; Alonso, O.; Cervera, J.; Such, E.; Silla, M. A.; Villaescusa, J. I.; Montoro, A.

2012-07-01

Study in vitro of origin radioprotective food the radioprotective effect in vitro of food borne substances studied is a first step in developing effective radioprotectors that can prevent radiation damage to healthy tissue., cannot forget that these studies must be accompanied by in vitro studies of toxicity and bioavailability to profile designing radioprotective substance.

Approaches and Challenges of Engineering Implantable Microelectromechanical Systems (MEMS Drug Delivery Systems for in Vitro and in Vivo Applications

Directory of Open Access Journals (Sweden)

Ken-Tye Yong

2012-11-01

Full Text Available Despite the advancements made in drug delivery systems over the years, many challenges remain in drug delivery systems for treating chronic diseases at the personalized medicine level. The current urgent need is to develop novel strategies for targeted therapy of chronic diseases. Due to their unique properties, microelectromechanical systems (MEMS technology has been recently engineered as implantable drug delivery systems for disease therapy. This review examines the challenges faced in implementing implantable MEMS drug delivery systems in vivo and the solutions available to overcome these challenges.

The cyber threat landscape: Challenges and future research directions

Science.gov (United States)

Gil, Santiago; Kott, Alexander; Barabási, Albert-László

2014-07-01

While much attention has been paid to the vulnerability of computer networks to node and link failure, there is limited systematic understanding of the factors that determine the likelihood that a node (computer) is compromised. We therefore collect threat log data in a university network to study the patterns of threat activity for individual hosts. We relate this information to the properties of each host as observed through network-wide scans, establishing associations between the network services a host is running and the kinds of threats to which it is susceptible. We propose a methodology to associate services to threats inspired by the tools used in genetics to identify statistical associations between mutations and diseases. The proposed approach allows us to determine probabilities of infection directly from observation, offering an automated high-throughput strategy to develop comprehensive metrics for cyber-security.

3D bioprinting: improving in vitro models of metastasis with heterogeneous tumor microenvironments

Directory of Open Access Journals (Sweden)

Jacob L. Albritton

2017-01-01

Full Text Available Even with many advances in treatment over the past decades, cancer still remains a leading cause of death worldwide. Despite the recognized relationship between metastasis and increased mortality rate, surprisingly little is known about the exact mechanism of metastatic progression. Currently available in vitro models cannot replicate the three-dimensionality and heterogeneity of the tumor microenvironment sufficiently to recapitulate many of the known characteristics of tumors in vivo. Our understanding of metastatic progression would thus be boosted by the development of in vitro models that could more completely capture the salient features of cancer biology. Bioengineering groups have been working for over two decades to create in vitro microenvironments for application in regenerative medicine and tissue engineering. Over this time, advances in 3D printing technology and biomaterials research have jointly led to the creation of 3D bioprinting, which has improved our ability to develop in vitro models with complexity approaching that of the in vivo tumor microenvironment. In this Review, we give an overview of 3D bioprinting methods developed for tissue engineering, which can be directly applied to constructing in vitro models of heterogeneous tumor microenvironments. We discuss considerations and limitations associated with 3D printing and highlight how these advances could be harnessed to better model metastasis and potentially guide the development of anti-cancer strategies.

3D bioprinting: improving in vitro models of metastasis with heterogeneous tumor microenvironments.

Science.gov (United States)

Albritton, Jacob L; Miller, Jordan S

2017-01-01

Even with many advances in treatment over the past decades, cancer still remains a leading cause of death worldwide. Despite the recognized relationship between metastasis and increased mortality rate, surprisingly little is known about the exact mechanism of metastatic progression. Currently available in vitro models cannot replicate the three-dimensionality and heterogeneity of the tumor microenvironment sufficiently to recapitulate many of the known characteristics of tumors in vivo Our understanding of metastatic progression would thus be boosted by the development of in vitro models that could more completely capture the salient features of cancer biology. Bioengineering groups have been working for over two decades to create in vitro microenvironments for application in regenerative medicine and tissue engineering. Over this time, advances in 3D printing technology and biomaterials research have jointly led to the creation of 3D bioprinting, which has improved our ability to develop in vitro models with complexity approaching that of the in vivo tumor microenvironment. In this Review, we give an overview of 3D bioprinting methods developed for tissue engineering, which can be directly applied to constructing in vitro models of heterogeneous tumor microenvironments. We discuss considerations and limitations associated with 3D printing and highlight how these advances could be harnessed to better model metastasis and potentially guide the development of anti-cancer strategies. © 2017. Published by The Company of Biologists Ltd.

Assessment of carbon sources on in vitro shoot regeneration in tomato

International Nuclear Information System (INIS)

Shah, S.H.; Jan, S.A.

2014-01-01

An innovative approach for in vitro shoot regeneration by both direct and indirect means was developed in three tomato genotypes culturing hypocotyls and leaf discs explants on MS and N6 basal media fortified with various concentrations of carbon sources (sucrose and sorbitol) individually, accumulatively and also in amalgamation with various plant growth regulators. No response of in vitro shoot regeneration was recorded in all the genotypes by the individual application of carbon sources in both MS and N6 basal media. On the other hand, their accumulative effect rapidly enhanced the in vitro shoot regeneration frequency in all the genotypes. The highest shoot organogenesis frequency (100, 99.00 and 97.69%) was recorded in Rio Grande, Roma and Moneymaker, respectively on MS medium fortified with carbon sources (30: 30 g/l) culturing hypocotyls. Supplementation of sucrose: sorbitol (30: 30 g/l) in N6 medium along with different PGRs (0.1 mg/l IAA, 1.0 mg/l ZEA and 2.0 mg/l BAP) produced the highest shoot regeneration frequency (96.33, 92.69 and 88.74%) in Roma, Rio Grande and Moneymaker culturing leaf discs. Our findings suggest an alternative approach as hormone-free protocol for in vitro shoot regeneration in tomato that would save the resources with regard to hormonal costs and time. (author)

China’s Foreign Direct Investments:Challenges of Due Diligence and Organizational Integration

Institute of Scientific and Technical Information of China (English)

ABDOL; S.SOOFI

2015-01-01

This paper critically reviews Chinese companies’ foreign direct investment practices of recent years. Using case studies involving overseas Greenfield as well as merger and acquisition(M&A;) of Chinese enterprises, we aim to draw lessons from these experiences. However, because of increasing importance of outbound acquisitions by Chinese companies, this paper focuses on Chinese M&A; activities. After presenting the theoretical discussions of post-acquisition organizational integration, this paper identifies factors that have contributed to less than expected performances of Chinese foreign investments. Three main factors are identified as the plausible causes of the less than satisfactory outcomes: inadequate due diligence, not considering political and country risks, and cultural differences. In all cases, inexperience of Chinese enterprises in foreign direct investment, either in Greenfield form or M&A;, has attributed to the problems. Therefore, summing the experiences of the Chinese enterprises that have foreign direct investment is essential for those Chinese investors that intend to invest overseas. Conduct of meaningful, in-depth due diligence before serious negotiations for investment or acquisition, inclusion of risk premium for political risk in cash flow analysis, and early post-merger integration planning are essential for avoidances of bitter outcomes many Chinese investors experienced overseas.

Root damage induced by intraosseous anesthesia?An in vitro investigation

OpenAIRE

Graetz, Christian; Fawzy-El-Sayed, Karim M.; Graetz, Nicole; D?rfer, Christof-Edmund

2012-01-01

Objectives: The principle of the intraosseous anesthesia (IOA) relies on the perforation of the cortical plate of the bone for direct application of the local anesthetic solution into the underlying cancellous structures. During this procedure, IOA needles might accidentally come in contact with the tooth roots. The aim of the current in vitro study was to examine the consequences of this ?worst case scenario? comparing five commercially available IOA systems. Material and Methods: Extracted ...

Software challenges in extreme scale systems

International Nuclear Information System (INIS)

Sarkar, Vivek; Harrod, William; Snavely, Allan E

2009-01-01

Computer systems anticipated in the 2015 - 2020 timeframe are referred to as Extreme Scale because they will be built using massive multi-core processors with 100's of cores per chip. The largest capability Extreme Scale system is expected to deliver Exascale performance of the order of 10 18 operations per second. These systems pose new critical challenges for software in the areas of concurrency, energy efficiency and resiliency. In this paper, we discuss the implications of the concurrency and energy efficiency challenges on future software for Extreme Scale Systems. From an application viewpoint, the concurrency and energy challenges boil down to the ability to express and manage parallelism and locality by exploring a range of strong scaling and new-era weak scaling techniques. For expressing parallelism and locality, the key challenges are the ability to expose all of the intrinsic parallelism and locality in a programming model, while ensuring that this expression of parallelism and locality is portable across a range of systems. For managing parallelism and locality, the OS-related challenges include parallel scalability, spatial partitioning of OS and application functionality, direct hardware access for inter-processor communication, and asynchronous rather than interrupt-driven events, which are accompanied by runtime system challenges for scheduling, synchronization, memory management, communication, performance monitoring, and power management. We conclude by discussing the importance of software-hardware co-design in addressing the fundamental challenges for application enablement on Extreme Scale systems.

Grand Challenges and Great Potential in Foreign Language Teaching and Learning

Science.gov (United States)

Hlas, Anne Cummings

2018-01-01

This article argues for the field of foreign languages to begin to identify and define our Grand Challenges, which are difficult yet solvable problems facing our field. Seeking answers to these challenges can provide new opportunities for collaboration and can spur new directions and innovation within language learning and teaching. Researchable…

Challenges of the banking consolidation in Nigeria | Oyefeso ...

African Journals Online (AJOL)

CBN) and the new mega banks may face as a consequence of the banking consolidation reforms in Nigeria. Surmounting these challenges will require CBN's commitment to implementing the Basel II directives and instituting adequate Act of ...

In Vitro Evaluation of Biofilm Dispersal as a Therapeutic Strategy To Restore Antimicrobial Efficacy

DEFF Research Database (Denmark)

Roizman, Dan; Vidaillac, Celine; Givskov, Michael

2017-01-01

As a proof-of-concept study, the direct impact of biofilm dispersal on the in vitro efficacy of imipenem and tobramycin was evaluated against 3-day-old biofilms of Pseudomonas aeruginosa. Arabinose induction of biofilm dispersal via activation of the phosphodiesterase YhjH in the P. aeruginosa en...

Engineered skeletal muscle tissue for soft robotics: fabrication strategies, current applications, and future challenges.

Science.gov (United States)

Duffy, Rebecca M; Feinberg, Adam W

2014-01-01

Skeletal muscle is a scalable actuator system used throughout nature from the millimeter to meter length scales and over a wide range of frequencies and force regimes. This adaptability has spurred interest in using engineered skeletal muscle to power soft robotics devices and in biotechnology and medical applications. However, the challenges to doing this are similar to those facing the tissue engineering and regenerative medicine fields; specifically, how do we translate our understanding of myogenesis in vivo to the engineering of muscle constructs in vitro to achieve functional integration with devices. To do this researchers are developing a number of ways to engineer the cellular microenvironment to guide skeletal muscle tissue formation. This includes understanding the role of substrate stiffness and the mechanical environment, engineering the spatial organization of biochemical and physical cues to guide muscle alignment, and developing bioreactors for mechanical and electrical conditioning. Examples of engineered skeletal muscle that can potentially be used in soft robotics include 2D cantilever-based skeletal muscle actuators and 3D skeletal muscle tissues engineered using scaffolds or directed self-organization. Integration into devices has led to basic muscle-powered devices such as grippers and pumps as well as more sophisticated muscle-powered soft robots that walk and swim. Looking forward, current, and future challenges include identifying the best source of muscle precursor cells to expand and differentiate into myotubes, replacing cardiomyocytes with skeletal muscle tissue as the bio-actuator of choice for soft robots, and vascularization and innervation to enable control and nourishment of larger muscle tissue constructs. © 2013 Wiley Periodicals, Inc.

In vivo and in vitro toxicological effects of titanium dioxide nanoparticles on small intestine

Science.gov (United States)

Tassinari, Roberta; La Rocca, Cinzia; Stecca, Laura; Tait, Sabrina; De Berardis, Barbara; Ammendolia, Maria Grazia; Iosi, Francesca; Di Virgilio, Antonio; Martinelli, Andrea; Maranghi, Francesca

2015-06-01

In European Union, titanium dioxide (TiO2) as bulk material is a food additive (E171) and - as nanoparticle (NP) - is used as a white pigment in several products (e.g. food, cosmetics, drugs). E171 contains approximately 36% of particles less than 100 nm in at least one dimension and TiO2 NP exposure is estimated fairly below 2.5 mg/person/day. The gastrointestinal tract is a route of entry for NPs, thus representing a potential target of effects. In in vivo study, the effects of TiO2 NP in adult rat small intestine have been evaluated by oral administration of 0 (CTRL), 1 and 2 mg/kg body weight per day - relevant to human dietary intake. Detailed quali/quantitative histopathological analyses were performed on CTRL and treated rat samples. Scanning electron microscopy (SEM) analysis was performed on small intestine. An in vitro study on Caco-2 cells was also used in order to evaluate the potential cytotoxic effects directly on enterocytes through the lactate dehydrogenase (LDH) assay. Suspensions of TiO2 NPs for in vitro and in vivo study were characterized by EM. Histomorphometrical data showed treatment-related changes of villus height and widths in male rats. Significantly different from CTRL decreased LDH levels in the medium were detected in vitro at 24h with 2.5, 5, 10 and 20 µg/cm2 levels of TiO2 NPs. SEM analysis showed no damaged areas. Overall the results showed that enterocytes may represent a target of TiO2 NP toxicity by direct exposure both in vivo and in vitro models.

In vivo and in vitro toxicological effects of titanium dioxide nanoparticles on small intestine

International Nuclear Information System (INIS)

Tassinari, Roberta; La Rocca, Cinzia; Tait, Sabrina; De Berardis, Barbara; Ammendolia, Maria Grazia; Iosi, Francesca; Di Virgilio, Antonio; Martinelli, Andrea; Maranghi, Francesca; Stecca, Laura

2014-01-01

In European Union, titanium dioxide (TiO 2 ) as bulk material is a food additive (E171) and - as nanoparticle (NP) - is used as a white pigment in several products (e.g. food, cosmetics, drugs). E171 contains approximately 36% of particles less than 100 nm in at least one dimension and TiO 2 NP exposure is estimated fairly below 2.5 mg/person/day. The gastrointestinal tract is a route of entry for NPs, thus representing a potential target of effects. In in vivo study, the effects of TiO 2 NP in adult rat small intestine have been evaluated by oral administration of 0 (CTRL), 1 and 2 mg/kg body weight per day - relevant to human dietary intake. Detailed quali/quantitative histopathological analyses were performed on CTRL and treated rat samples. Scanning electron microscopy (SEM) analysis was performed on small intestine. An in vitro study on Caco-2 cells was also used in order to evaluate the potential cytotoxic effects directly on enterocytes through the lactate dehydrogenase (LDH) assay. Suspensions of TiO 2 NPs for in vitro and in vivo study were characterized by EM. Histomorphometrical data showed treatment-related changes of villus height and widths in male rats. Significantly different from CTRL decreased LDH levels in the medium were detected in vitro at 24h with 2.5, 5, 10 and 20 µg/cm 2 levels of TiO 2 NPs. SEM analysis showed no damaged areas. Overall the results showed that enterocytes may represent a target of TiO 2 NP toxicity by direct exposure both in vivo and in vitro models

Topology Control in Aerial Multi-Beam Directional Networks

Science.gov (United States)

2017-04-24

Topology Control in Aerial Multi-Beam Directional Networks Brian Proulx, Nathaniel M. Jones, Jennifer Madiedo, Greg Kuperman {brian.proulx, njones...significant interference. Topology control (i.e., selecting a subset of neighbors to communicate with) is vital to reduce the interference. Good topology ...underlying challenges to topology control in multi-beam direction networks. Two topology control algorithms are developed: a centralized algorithm

Presidential Green Chemistry Challenge: 2006 Greener Reaction Conditions Award

Science.gov (United States)

Presidential Green Chemistry Challenge 2006 award winner, Codexis, directed the evolution of three designer enzymes to produce the key chiral building block for atorvastatin, the active ingredient in the drug Lipitor.

Combination of siRNA-directed Kras oncogene silencing and arsenic-induced apoptosis using a nanomedicine strategy for the effective treatment of pancreatic cancer.

Science.gov (United States)

Zeng, Linjuan; Li, Jingguo; Wang, Yong; Qian, Chenchen; Chen, Yinting; Zhang, Qiubo; Wu, Wei; Lin, Zhong; Liang, Jianzhong; Shuai, Xintao; Huang, Kaihong

2014-02-01

The synergetic inhibitory effects on human pancreatic cancer by nanoparticle-mediated siRNA and arsenic therapy were investigated both in vitro and in vivo. Poly(ethylene glycol)-block-poly(L-lysine) were prepared to form siRNA-complexed polyplex and poly(ethylene glycol)-block-poly(DL-lactide) were prepared to form arsenic-encapsulated vesicle, respectively. Down-regulation of the mutant Kras gene by siRNA caused defective abilities of proliferation, clonal formation, migration, and invasion of pancreatic cancer cells, as well as cell cycle arrest at the G0/G1 phase, which substantially enhanced the apoptosis-inducing effect of arsenic administration. Consequently, co-administration of the two nanomedicines encapsulating siRNA or arsenic showed ideal tumor growth inhibition both in vitro and in vivo as a result of synergistic effect of the siRNA-directed Kras oncogene silencing and arsenic-induced cell apoptosis. These results suggest that the combination of mutant Kras gene silencing and arsenic therapy using nanoparticle-mediated delivery strategy is promising for pancreatic cancer treatment. Treatment of pancreatic cancer remains a major challenge. These authors demonstrate a method that combines a siRNA-based Kras silencing with arsenic delivery to pancreatic cancer cells using nanoparticles, resulting in enhanced apoptosis induction in the treated cells. © 2013.

Potential theory for directed networks.

Directory of Open Access Journals (Sweden)

Qian-Ming Zhang

Full Text Available Uncovering factors underlying the network formation is a long-standing challenge for data mining and network analysis. In particular, the microscopic organizing principles of directed networks are less understood than those of undirected networks. This article proposes a hypothesis named potential theory, which assumes that every directed link corresponds to a decrease of a unit potential and subgraphs with definable potential values for all nodes are preferred. Combining the potential theory with the clustering and homophily mechanisms, it is deduced that the Bi-fan structure consisting of 4 nodes and 4 directed links is the most favored local structure in directed networks. Our hypothesis receives strongly positive supports from extensive experiments on 15 directed networks drawn from disparate fields, as indicated by the most accurate and robust performance of Bi-fan predictor within the link prediction framework. In summary, our main contribution is twofold: (i We propose a new mechanism for the local organization of directed networks; (ii We design the corresponding link prediction algorithm, which can not only testify our hypothesis, but also find out direct applications in missing link prediction and friendship recommendation.

Potential Theory for Directed Networks

Science.gov (United States)

Zhang, Qian-Ming; Lü, Linyuan; Wang, Wen-Qiang; Zhou, Tao

2013-01-01

Uncovering factors underlying the network formation is a long-standing challenge for data mining and network analysis. In particular, the microscopic organizing principles of directed networks are less understood than those of undirected networks. This article proposes a hypothesis named potential theory, which assumes that every directed link corresponds to a decrease of a unit potential and subgraphs with definable potential values for all nodes are preferred. Combining the potential theory with the clustering and homophily mechanisms, it is deduced that the Bi-fan structure consisting of 4 nodes and 4 directed links is the most favored local structure in directed networks. Our hypothesis receives strongly positive supports from extensive experiments on 15 directed networks drawn from disparate fields, as indicated by the most accurate and robust performance of Bi-fan predictor within the link prediction framework. In summary, our main contribution is twofold: (i) We propose a new mechanism for the local organization of directed networks; (ii) We design the corresponding link prediction algorithm, which can not only testify our hypothesis, but also find out direct applications in missing link prediction and friendship recommendation. PMID:23408979

Nanoscale Science, Engineering and Technology Research Directions

Energy Technology Data Exchange (ETDEWEB)

Lowndes, D. H.; Alivisatos, A. P.; Alper, M.; Averback, R. S.; Jacob Barhen, J.; Eastman, J. A.; Imre, D.; Lowndes, D. H.; McNulty, I.; Michalske, T. A.; Ho, K-M; Nozik, A. J.; Russell, T. P.; Valentin, R. A.; Welch, D. O.; Barhen, J.; Agnew, S. R.; Bellon, P.; Blair, J.; Boatner, L. A.; Braiman, Y.; Budai, J. D.; Crabtree, G. W.; Feldman, L. C.; Flynn, C. P.; Geohegan, D. B.; George, E. P.; Greenbaum, E.; Grigoropoulos, C.; Haynes, T. E.; Heberlein, J.; Hichman, J.; Holland, O. W.; Honda, S.; Horton, J. A.; Hu, M. Z.-C.; Jesson, D. E.; Joy, D. C.; Krauss, A.; Kwok, W.-K.; Larson, B. C.; Larson, D. J.; Likharev, K.; Liu, C. T.; Majumdar, A.; Maziasz, P. J.; Meldrum, A.; Miller, J. C.; Modine, F. A.; Pennycook, S. J.; Pharr, G. M.; Phillpot, S.; Price, D. L.; Protopopescu, V.; Poker, D. B.; Pui, D.; Ramsey, J. M.; Rao, N.; Reichl, L.; Roberto, J.; Saboungi, M-L; Simpson, M.; Strieffer, S.; Thundat, T.; Wambsganss, M.; Wendleken, J.; White, C. W.; Wilemski, G.; Withrow, S. P.; Wolf, D.; Zhu, J. H.; Zuhr, R. A.; Zunger, A.; Lowe, S.

1999-01-01

This report describes important future research directions in nanoscale science, engineering and technology. It was prepared in connection with an anticipated national research initiative on nanotechnology for the twenty-first century. The research directions described are not expected to be inclusive but illustrate the wide range of research opportunities and challenges that could be undertaken through the national laboratories and their major national scientific user facilities with the support of universities and industry.

Encapsulation of hydrophobic allergens into nanoparticles improves the in vitro immunological diagnosis of allergic contact dermatitis.

Science.gov (United States)

Cortial, Angèle; Nosbaum, Audrey; Rozières, Aurore; Baeck, Marie; de Montjoye, Laurence; Grande, Sophie; Briançon, Stéphanie; Nicolas, Jean-François; Vocanson, Marc

2015-05-01

The diagnosis of allergic contact dermatitis (ACD) relies on in vivo patch testing. In vitro immunological assays based on the characterization of circulating allergen-specific memory T cells represent a promising alternative to patch testing. However, their development is hampered by the technical challenge of assessing hydrophobic allergens in serum-based assays. In this study, we show that the encapsulation of fragrance mix 1 (FMI, a mixture of 8 hydrophobic allergens) into poly-ε-caprolactone nanoparticle (NP) vectors: (1) dramatically increases the solubilization of allergens in conventional cell culture media and (2) allows for a robust in vitro reactivation of allergen-specific T cells in large numbers of fragrance allergic patients. Therefore, the encapsulation of hydrophobic allergens into NP vectors opens new avenues to improve the in vitro immunobiological diagnosis of ACD. Allergic Contact Dermatitis (ACD) is a delayed-type hypersensivity reaction prevalent in many individuals. Currently, skin patch testing has been the mainstay for diagnosis clinically. In this study, the authors described an improvement to in vitro immunological assays measuring circulating allergen-specific memory T cells, using nanoparticle vectors. The positive data might provide an exciting alternative to current practice of patch-testing. Copyright © 2015 Elsevier Inc. All rights reserved.

Induction of miR-137 by Isorhapontigenin (ISO) Directly Targets Sp1 Protein Translation and Mediates Its Anticancer Activity Both In Vitro and In Vivo.

Science.gov (United States)

Zeng, Xingruo; Xu, Zhou; Gu, Jiayan; Huang, Haishan; Gao, Guangxun; Zhang, Xiaoru; Li, Jingxia; Jin, Honglei; Jiang, Guosong; Sun, Hong; Huang, Chuanshu

2016-03-01

Our recent studies found that isorhapontigenin (ISO) showed a significant inhibitory effect on human bladder cancer cell growth, accompanied with cell-cycle G0-G1 arrest as well as downregulation of Cyclin D1 expression at transcriptional level via inhibition of Sp1 transactivation in bladder cancer cells. In the current study, the potential ISO inhibition of bladder tumor formation has been explored in a xenograft nude mouse model, and the molecular mechanisms underlying ISO inhibition of Sp1 expression and anticancer activities have been elucidated both in vitro and in vivo. Moreover, the studies demonstrated that ISO treatment induced the expression of miR-137, which in turn suppressed Sp1 protein translation by directly targeting Sp1 mRNA 3'-untranslated region (UTR). Similar to ISO treatment, ectopic expression of miR-137 alone led to G0-G1 cell growth arrest and inhibition of anchorage-independent growth in human bladder cancer cells, which could be completely reversed by overexpression of GFP-Sp1. The inhibition of miR-137 expression attenuated ISO-induced inhibition of Sp1/Cyclin D1 expression, induction of G0-G1 cell growth arrest, and suppression of cell anchorage-independent growth. Taken together, our studies have demonstrated that miR-137 induction by ISO targets Sp1 mRNA 3'-UTR and inhibits Sp1 protein translation, which consequently results in reduction of Cyclin D1 expression, induction of G0-G1 growth arrest, and inhibition of anchorage-independent growth in vitro and in vivo. Our results have provided novel insights into understanding the anticancer activity of ISO in the therapy of human bladder cancer. ©2016 American Association for Cancer Research.

MiR-34a-3p alters proliferation and apoptosis of meningioma cells in vitro and is directly targeting SMAD4, FRAT1 and BCL2

Science.gov (United States)

Werner, Tamara V.; Hart, Martin; Nickels, Ruth; Kim, Yoo-Jin; Menger, Michael D.; Bohle, Rainer M.; Keller, Andreas; Ludwig, Nicole; Meese, Eckart

2017-01-01

Micro (mi)RNAs are short, noncoding RNAs and deregulation of miRNAs and their targets are implicated in tumor generation and progression in many cancers. Meningiomas are mostly benign, slow growing tumors of the central nervous system with a small percentage showing a malignant phenotype. Following in silico prediction of potential targets of miR-34a-3p, SMAD4, FRAT1, and BCL2 have been confirmed as targets by dual luciferase assays with co-expression of miR-34a-3p and reporter gene constructs containing the respective 3'UTRs. Disruption of the miR-34a-3p binding sites in the 3'UTRs resulted in loss of responsiveness to miR-34a-3p overexpression. In meningioma cells, overexpression of miR-34a-3p resulted in decreased protein levels of SMAD4, FRAT1 and BCL2, while inhibition of miR-34a-3p led to increased levels of these proteins as confirmed by Western blotting. Furthermore, deregulation of miR-34a-3p altered cell proliferation and apoptosis of meningioma cells in vitro. We show that SMAD4, FRAT1 and BCL2 are direct targets of miR-34a-3p and that deregulation of miR-34a-3p alters proliferation and apoptosis of meningioma cells in vitro. As part of their respective signaling pathways, which are known to play a role in meningioma genesis and progression, deregulation of SMAD4, FRAT1 and BCL2 might contribute to the aberrant activation of these signaling pathways leading to increased proliferation and inhibition of apoptosis in meningiomas. PMID:28340489

Laser-assisted cartilage reshaping: in vitro and in vivo animal studies

Science.gov (United States)

Wang, Zhi; Pankratov, Michail M.; Perrault, Donald F., Jr.; Shapshay, Stanley M.

1995-05-01

Correction of cartilaginous defects in the head and neck area remains a challenge for the surgeon. This study investigated a new technique for laser-assisted cartilage reshaping. The pulsed 1.44 micrometers Nd:YAG laser was used in vitro and in vivo experiments to irradiate cartilage to change it's shape without carbonization or vaporization of tissue. Two watts of average power in non contact manner was used to irradiate and reshape the cartilage. The extracted reshaped cartilage specimens underwent testing of elastic force with a computer assisted measurement system that recorded the changes in elastic force in the specimens from 1 hr to 11 days post-irradiation. An animal model of defective tracheal cartilage (collapsed tracheal wall) was created, allowed to heal for 6 weeks and then corrected endoscopically with the laser-assisted technique. The results of the in vitro and in vivo investigations demonstrated that it was possible to alter the cartilage and that cartilage would retain its new shape. The clinical significance of the technique is evident and warrants further animal studies and clinical trials.

Evaluación de la eficacia in vitro e in vivo del tratamiento antimicrobiano combinado por el tablero de damas Effectiveness of a combined antimicrobial treatment by means of the draughtboard. In vitro and in vivo assessment

Directory of Open Access Journals (Sweden)

María Espino Hernández

2009-08-01

Full Text Available El presente trabajo tuvo como objetivo, evaluar la eficacia clínica del tratamiento antimicrobiano combinado mediante pruebas de sinergismo in vitro y determinar la eficacia del procedimiento empleado para orientar la aplicación de estas formas de tratamiento en el paciente con sepsis por bacterias con resistencia variada. Se evaluó el tratamiento aplicado a 163 recién nacidos con sepsis corroborada por hemocultivo positivo en el Hospital Ginecobstétrico Universitario "América Arias", durante el periodo de enero de 1993 a diciembre de 2000, por el método tablero de damas (checkerboard en placas de microtitulación. Las combinaciones de fármacos sinérgicas in vitro presentaron una alta probabilidad de eficacia clínica, independiente del patrón de susceptibilidad del microorganismo aun frente a cepas resistentes a los 2 antimicrobianos utilizados en el tratamiento. Todos los resultados antagónicos in vitro se correspondieron con un fallo clínico terapéutico. El procedimiento utilizado puede constituir una valiosa herramienta para orientar la terapéutica en pacientes con sepsis por microorganismos resistentes.The aim of present paper was to evaluate clinical effectiveness of combined antimicrobial treatment by means of in vitro synergism tests and to determine effectiveness of used procedure to direct applications of these treatment strategies in patient presenting with sepsis from varied resistance bacteria. Treatment applied was assessed in 163 newborn patients presenting with sepsis from positive hemoculture in "America Arias" University Gynecology-Obstetrics Hospital from January 1993 to December 2000, by checkerboard method in microtritate plates. In vitro synergic drug combinations showed a high probability of clinical effectiveness, independently of susceptibility pattern of microorganism event versus resistant strains to both antimicrobials used in treatment. All in vitro antagonistic results correspond both with a

Opportunities and Challenges with Digital Open Badges

Science.gov (United States)

Farmer, Tadd; West, Richard E.

2016-01-01

With increasing interest in competency and outcome-based education, and the blending of formal and informal learning, there is increasing need for credentials to match these learning paradigms. In this article, the authors discuss the benefits, challenges, and potential future directions for open digital badges--one potential alternative…

Acclimatization of in Vitro-derived Dendrobium

Directory of Open Access Journals (Sweden)

Jaime A. Teixeira da Silva

2017-05-01

Full Text Available The successful ex vitro establishment of Dendrobium plantlets raised in vitro determines the quality of the end product (cut flowers or potted plants in commercial production for economic gain. When in vitro Dendrobium plantlets are transplanted from the culture room to greenhouse conditions, they may desiccate or wilt rapidly and can die as a result of changes in the environment, unless substantial precautions are taken to adapt plantlets to a new environment. The acclimatization of in vitro-grown Dendrobium plantlets to an ex vitro environment by gradually weaning them towards ambient relative humidity and light levels facilitates better survival of young and physiologically sensitive plantlets. Dendrobium plantlets raised in vitro must thus undergo a period of acclimatization or transitional development to correct anatomical abnormalities and to enhance their physiological performance to ensure survival under ex vitro conditions. The most common approach to improve the survival of Dendrobium plantlets upon transfer to an ex vitro environment is their gradual adaptation to that environment. Under such conditions, plants convert rapidly from a heterotrophic or photomixotrophic state to an autotrophic growth, develop a fully functional root system, and better control their stomatal and cuticular transpiration. Gradual adaptation is carried out in a greenhouse by decreasing relative humidity using fog or mist chambers and by increasing light intensity using shading techniques. This review details the acclimatization and ex vitro survival of Dendrobium plants produced in vitro. This advice is also useful for other orchids.

Research Challenges in Financial Data Modeling and Analysis.

Science.gov (United States)

Alexander, Lewis; Das, Sanjiv R; Ives, Zachary; Jagadish, H V; Monteleoni, Claire

2017-09-01

Significant research challenges must be addressed in the cleaning, transformation, integration, modeling, and analytics of Big Data sources for finance. This article surveys the progress made so far in this direction and obstacles yet to be overcome. These are issues that are of interest to data-driven financial institutions in both corporate finance and consumer finance. These challenges are also of interest to the legal profession as well as to regulators. The discussion is relevant to technology firms that support the growing field of FinTech.

In vitro large-scale experimental and theoretical studies for the realization of bi-directional brain-prostheses.

Science.gov (United States)

Bonifazi, Paolo; Difato, Francesco; Massobrio, Paolo; Breschi, Gian L; Pasquale, Valentina; Levi, Timothée; Goldin, Miri; Bornat, Yannick; Tedesco, Mariateresa; Bisio, Marta; Kanner, Sivan; Galron, Ronit; Tessadori, Jacopo; Taverna, Stefano; Chiappalone, Michela

2013-01-01

Brain-machine interfaces (BMI) were born to control "actions from thoughts" in order to recover motor capability of patients with impaired functional connectivity between the central and peripheral nervous system. The final goal of our studies is the development of a new proof-of-concept BMI-a neuromorphic chip for brain repair-to reproduce the functional organization of a damaged part of the central nervous system. To reach this ambitious goal, we implemented a multidisciplinary "bottom-up" approach in which in vitro networks are the paradigm for the development of an in silico model to be incorporated into a neuromorphic device. In this paper we present the overall strategy and focus on the different building blocks of our studies: (i) the experimental characterization and modeling of "finite size networks" which represent the smallest and most general self-organized circuits capable of generating spontaneous collective dynamics; (ii) the induction of lesions in neuronal networks and the whole brain preparation with special attention on the impact on the functional organization of the circuits; (iii) the first production of a neuromorphic chip able to implement a real-time model of neuronal networks. A dynamical characterization of the finite size circuits with single cell resolution is provided. A neural network model based on Izhikevich neurons was able to replicate the experimental observations. Changes in the dynamics of the neuronal circuits induced by optical and ischemic lesions are presented respectively for in vitro neuronal networks and for a whole brain preparation. Finally the implementation of a neuromorphic chip reproducing the network dynamics in quasi-real time (10 ns precision) is presented.

In vitro large-scale experimental and theoretical studies for the realization of bi-directional brain-prostheses

Directory of Open Access Journals (Sweden)

Paolo eBonifazi

2013-03-01

Full Text Available Brain-machine interfaces (BMI were born to control ‘actions from thoughts’ in order to recover motor capability of patients with impaired functional connectivity between the central and peripheral nervous system. The final goal of our studies is the development of a new proof-of-concept BMI - a neuromorphic chip for brain repair - to reproduce the functional organization of a damaged part of the central nervous system. To reach this ambitious goal, we implemented a multidisciplinary ‘bottom-up’ approach in which in vitro networks are the paradigm for the development of an in silico model to be incorporated into a neuromorphic device. In this paper we present the overall strategy and focus on the different building blocks of our studies: (i the experimental characterization and modeling of ‘finite size networks’ which represent the smallest and most general self-organized circuits capable of generating spontaneous collective dynamics; (ii the induction of lesions in neuronal networks and the whole brain preparation with special attention on the impact on the functional organization of the circuits; (iii the first production of a neuromorphic chip able to implement a real-time model of neuronal networks. A dynamical characterization of the finite size circuits with single cell resolution is provided. A neural network model based on Izhikevich neurons was able to replicate the experimental observations. Changes in the dynamics of the neuronal circuits induced by optical and ischemic lesions are presented respectively for in vitro neuronal networks and for a whole brain preparation. Finally the implementation of a neuromorphic chip reproducing the network dynamics in quasi-real time (10 ns precision is presented.

Home telemonitoring of vital signs--technical challenges and future directions.

Science.gov (United States)

Celler, Branko G; Sparks, Ross S

2015-01-01

The telemonitoring of vital signs from the home is an essential element of telehealth services for the management of patients with chronic conditions, such as congestive heart failure (CHF), chronic obstructive pulmonary disease (COPD), diabetes, or poorly controlled hypertension. Telehealth is now being deployed widely in both rural and urban settings, and in this paper, we discuss the contribution made by biomedical instrumentation, user interfaces, and automated risk stratification algorithms in developing a clinical diagnostic quality longitudinal health record at home. We identify technical challenges in the acquisition of high-quality biometric signals from unsupervised patients at home, identify new technical solutions and user interfaces, and propose new measurement modalities and signal processing techniques for increasing the quality and value of vital signs monitoring at home. We also discuss use of vital signs data for the automated risk stratification of patients, so that clinical resources can be targeted to those most at risk of unscheduled admission to hospital. New research is also proposed to integrate primary care, hospital, personal genomic, and telehealth electronic health records, and apply predictive analytics and data mining for enhancing clinical decision support.

Mental health literacy as theory: current challenges and future directions.

Science.gov (United States)

Spiker, Douglas A; Hammer, Joseph H

2018-02-13

Mental health literacy (MHL) is one increasingly researched factor thought to influence mental health behaviors. Researchers have argued for expanding the definition of MHL to include additional constructs, but no consensus has yet been reached on what constructs should be included as part of MHL. The purpose of this paper is to (i) elucidate how the expansion of the MHL construct has impeded the growth of MHL research and (ii) through the lens of construct and theory development, highlight how these challenges might be remedied. An inclusive search of the literature was undertaken to identify MHL studies. The principles of construct and theory development guided a critical analysis of MHL. The review of the literature found that MHL violates many principles of what constitutes an acceptable construct definition. To address these concerns, we proposed conceptualizing MHL as a theory and recommended principles of theory development that should be taken into consideration. A theory of MHL can guide future researchers to clearly delineate important constructs and their interrelationships. For practitioners, a theory of MHL can help inform how to improve MHL at both the individual and community level.

Prophylactic Nipple-Sparing Mastectomy and Direct-to-Implant Reconstruction of the Large and Ptotic Breast: Is Preshaping of the Challenging Breast a Key to Success?

Science.gov (United States)

Gunnarsson, Gudjon L; Bille, Camilla; Reitsma, Laurens C; Wamberg, Peter; Thomsen, Jørn Bo

2017-09-01

Nipple-sparing mastectomy with simultaneous hammock technique direct-to-implant reconstruction is increasingly offered to patients opting for risk-reducing mastectomy. Despite this promising method, patients with macromastia and ptotic breasts remain a challenging group to treat satisfactorily and more often end up undergoing a difficult corrective procedure and experience an unacceptably high rate of failed reconstruction. The authors examined whether targeted preshaping mastopexy/reduction could prepare these patients for a successful nipple-sparing mastectomy/direct-to-implant reconstruction. Patients seeking risk-reducing nipple-sparing mastectomy/direct-to-implant reconstruction at the authors' institutions deemed unfit for a one-stage procedure based on their previous experience were offered a targeted two-stage, risk-reducing mastopexy/reduction followed by a delayed secondary nipple-sparing mastectomy and direct-to-implant reconstruction. Patients were followed up at 3 weeks and 6 or 12 months. Forty-four reconstructions were performed in 22 patients aged 43 years (range, 26 to 57 years). All 44 procedures were completed successfully without any failure or nipple-areola complex losses. Patients' median body mass index was 30 kg/m (range, 22 to 44 kg/m). Six patients were smokers and one had hypertension. Two patients underwent reoperation because of hematoma and fat necrosis. The authors' results demonstrate that a targeted preshaping mastopexy/reduction followed by nipple-sparing mastectomy/direct-to-implant reconstruction can be safely planned in women who opt for a risk-reducing mastectomy and can be performed successfully with a 3- to 4-month time span between operations. On the basis of these results and the superior cosmetic outcome, the two-stage approach has become the authors' standard of care in all such settings. Therapeutic, IV.

Rules of Engagement: Toward an Analysis of Staff Responses to Challenging Behavior.

Science.gov (United States)

Hastings, Richard P.; Remington, Bob

1994-01-01

This article reviews literature on the responses of direct care staff to challenging behaviors of individuals with mental retardation. The paper constructs a behavior analytic description of the functions of care staff behavior in relation to their clients' challenging behaviors, draws a distinction between contingency-shaped and rule-governed…

Employing in vitro directed molecular evolution for the selection of α-amylase variant inhibitors with activity toward cotton boll weevil enzyme.

Science.gov (United States)

da Silva, Maria Cristina Mattar; Del Sarto, Rafael Perseghini; Lucena, Wagner Alexandre; Rigden, Daniel John; Teixeira, Fabíola Rodrigues; Bezerra, Caroline de Andrade; Albuquerque, Erika Valéria Saliba; Grossi-de-Sa, Maria Fatima

2013-09-20

Numerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of α-amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, α-amylase inhibitors (α-AIs) represent a powerful tool to apply in the control of insect pests. Here, we applied DNA shuffling and phage display techniques and obtained a combinatorial library containing 10⁸ α-AI variant forms. From this library, variants were selected exhibiting in vitro affinity for cotton boll weevil α-amylases. Twenty-six variant sequences were cloned into plant expression vectors and expressed in Arabidopsis thaliana. Transformed plant extracts were assayed in vitro to select specific and potent α-amylase inhibitors against boll weevil amylases. While the wild type inhibitors, used to create the shuffled library, did not inhibit the A. grandis α-amylases, three α-AI mutants, named α-AIC3, α-AIA11 and α-AIG4 revealed high inhibitory activities against A. grandis α-amylases in an in vitro assay. In summary, data reported here shown the potential biotechnology of new α-AI variant genes for cotton boll weevil control. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

In vitro remineralization of in vivo and in vitro formed enamel lesions

NARCIS (Netherlands)

Iijima, Y; Takagi, O; Ruben, J; Arends, J

1999-01-01

Thin sections of natural white spot enamel lesions (WS) and of artificial in vitro lesions (VL) were remineralized simultaneously in vitro. The sections, clamped in a PMMA holder, were microradiographed at baseline and after remineralization in a calcium- and phosphate-containing solution (pH = 7.0;

Indigenous traditional medicine: in vitro anti-giardial activity of plants used in the treatment of diarrhea.

Science.gov (United States)

Brandelli, Clara Lia Costa; Giordani, Raquel Brandt; De Carli, Geraldo Attilio; Tasca, Tiana

2009-06-01

The ethnopharmacology for treatment of parasitic infections facilitates and directs the search for new chemical entities. In this direction, this study evaluated the cytotoxicity in vitro, against trophozoites of Giardia lamblia, of aqueous extracts of leaves Achyrocline satureioides (Lam.) DC., barks of Eugenia uniflora L., aerial parts of Foeniculum vulgare Miller, and barks of Psidium guajava L. These plants are traditionally used for the treatment of diarrhea by the indigenous population Mbyá-Guarani, located at the Lomba do Pinheiro, Porto Alegre, Rio Grande do Sul, Brazil. The in vitro susceptibility qualitative analysis of G. lamblia to aqueous extracts was performed in serial dilutions from 2.5 to 0.02 mg/mL. Results revealed the minimal inhibitory concentrations: 0.313 mg/mL for A. satureioides and E. uniflora, 0.02 mg/mL for P. guajava, and F. vulgare did not present any cytotoxic effect. Quantitative assays of viable trophozoites, showed that A. satureioides presented the highest cytotoxic effect (93.5%), followed by P. guajava (82.2%), and E. uniflora (67.3%). Indigenous Guarani use mainly A. satureioides for the treatment of diarrhea, revealing the conformity with results obtained in vitro. Bioguided assays are necessary to identify the compounds responsible for the activity of the aqueous extract of A. satureioides.

Double pharmacological challenge on repolarization opens new avenues for drug safety research

DEFF Research Database (Denmark)

Thomsen, Morten Bækgaard

2007-01-01

pointes (TdP) arrhythmia. Both the pharmaceutical industry and the regulatory bodies are neglecting the available proarrhythmia models. In vitro studies have suggested that combined pharmacological hits on repolarization will produce a superior substrate for in vivo proarrhythmia, compared to the single......-drug assessment. By using consecutive pharmacological challenges, a simple model is proposed, in which combinatorial pharmacology is employed to provoke TdP in the conscious dog. The pharmaceutical industry interested in evaluating the proarrhythmic potential of their present and future drugs now has a simple...

Facebook marketing for a direct selling company

OpenAIRE

Egeberg, Chenette

2015-01-01

The purpose of the thesis was to provide recommendations for how the studied company could improve their Facebook marketing activities. The company was the Danish subsidiary of an multinational direct selling company of health and beauty products. The following research questions were devised: 1) What are the constraints and challenges for LR Denmark’s facebook marketing as a subsidiary of a Multinational Corporation and as a Direct Selling Company? 2) How can LR Denmark improve the...

Semiconductor product analysis challenges based on the 1999 ITRS

International Nuclear Information System (INIS)

Joseph, Thomas W.; Anderson, Richard E.; Gilfeather, Glen; LeClaire, Carole; Yim, Daniel

2001-01-01

One of the most significant challenges for technology characterization and failure analysis is to keep instrumentation and techniques in step with the development of technology itself. Not only are dimensions shrinking and new materials being employed, but the rate of change is increasing. According to the 1999 International Technology Roadmap for Semiconductors, 'The number and difficulty of the technical challenges continue to increase as technology moves forward'. It could be argued that technology cannot be developed without appropriate analytical techniques; nevertheless while much effort is being directed at materials and processes, only a small proportion is being directed at analysis. Whereas previous versions of the Semiconductor Industry Association roadmap contained a small number of implicit references to characterization and analysis, the 1999 ITRS contains many explicit references. It is clear that characterization is now woven through the roadmap, and technology developers in all areas appreciate the fact that new instrumentation and techniques will be required to sustain the rate of development the semiconductor industry has seen in recent years. Late in 1999, a subcommittee of the Sematech Product Analysis Forum (PAF) reviewed the ITRS and identified a 'top-ten' list of challenges which the failure analysis community will face as present technologies are extended and future technologies are developed. This paper discusses the PAF top-ten list of challenges, which is based primarily on the Difficult Challenges tables from each ITRS working group. Eight of the top-ten are challenges of significant technical magnitude; only two could be considered non-technical in nature. Most of these challenges cut across several working group areas and could be considered common threads in the roadmap, ranging from fault simulation and modeling to imaging small features, from electrical defect isolation to deprocessing. While evolutionary changes can be anticipated

African Languages in a Digital Age: Challenges and Opportunities ...

International Development Research Centre (IDRC) Digital Library (Canada)

2010-05-01

May 1, 2010 ... African Languages in a Digital Age: Challenges and Opportunities for Indigenous Language Computing. Book cover African ... Sign up now for IDRC news and views sent directly to your inbox each month. Subscribe.

In vitro production of small ruminant embryos: late improvements and further research.

Science.gov (United States)

de Souza-Fabjan, Joanna Maria Gonçalves; Panneau, Barbara; Duffard, Nicolas; Locatelli, Yann; de Figueiredo, José Ricardo; Freitas, Vicente José de Figueirêdo; Mermillod, Pascal

2014-06-01

Beyond the potential use of in vitro production of embryos (IVP) in breeding schemes, embryos are also required for the establishment of new biotechnologies such as cloning and transgenesis. Additionally, the knowledge of oocyte and embryo physiology acquired through IVP techniques may stimulate the further development of other techniques such as marker assisted and genomic selection of preimplantation embryos, and also benefit assisted procreation in human beings. Efficient in vitro embryo production is currently a major objective for livestock industries, including small ruminants. The heterogeneity of oocytes collected from growing follicles by laparoscopic ovum pick up or in ovaries of slaughtered females, remains an enormous challenge for IVM success, and still limits the rate of embryo development. In addition, the lower quality of the IVP embryos, compared with their in vivo-derived counterparts, translates into poor cryosurvival, which restricts the wider use of this promising technology. Therefore, many studies have been reported in an attempt to determine the most suitable conditions for IVM, IVF, and in vitro development to maximize embryo production rate and quality. This review aims to present the current panorama of IVP production in small ruminants, describing important steps for its success, reporting the recent advances and also the main obstacles identified for its improvement and dissemination. Copyright © 2014 Elsevier Inc. All rights reserved.

Recommended Research Directions for Improving the Validation of Complex Systems Models.

Energy Technology Data Exchange (ETDEWEB)

Vugrin, Eric D. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Trucano, Timothy G. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Swiler, Laura Painton [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Finley, Patrick D. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Flanagan, Tatiana Paz [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Naugle, Asmeret Bier [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Tsao, Jeffrey Y. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Verzi, Stephen Joseph [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2017-02-01

Improved validation for models of complex systems has been a primary focus over the past year for the Resilience in Complex Systems Research Challenge. This document describes a set of research directions that are the result of distilling those ideas into three categories of research -- epistemic uncertainty, strong tests, and value of information. The content of this document can be used to transmit valuable information to future research activities, update the Resilience in Complex Systems Research Challenge's roadmap, inform the upcoming FY18 Laboratory Directed Research and Development (LDRD) call and research proposals, and facilitate collaborations between Sandia and external organizations. The recommended research directions can provide topics for collaborative research, development of proposals, workshops, and other opportunities.

Synthesis of milligram quantities of proteins using a reconstituted in vitro protein synthesis system.

Science.gov (United States)

Kazuta, Yasuaki; Matsuura, Tomoaki; Ichihashi, Norikazu; Yomo, Tetsuya

2014-11-01

In this study, the amount of protein synthesized using an in vitro protein synthesis system composed of only highly purified components (the PURE system) was optimized. By varying the concentrations of each system component, we determined the component concentrations that result in the synthesis of 0.38 mg/mL green fluorescent protein (GFP) in batch mode and 3.8 mg/mL GFP in dialysis mode. In dialysis mode, protein concentrations of 4.3 and 4.4 mg/mL were synthesized for dihydrofolate reductase and β-galactosidase, respectively. Using the optimized system, the synthesized protein represented 30% (w/w) of the total protein, which is comparable to the level of overexpressed protein in Escherichia coli cells. This optimized reconstituted in vitro protein synthesis system may potentially be useful for various applications, including in vitro directed evolution of proteins, artificial cell assembly, and protein structural studies. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Sex in a test tube: testing the benefits of in vitro recombination.

Science.gov (United States)

Pesce, Diego; Lehman, Niles; de Visser, J Arjan G M

2016-10-19

The origin and evolution of sex, and the associated role of recombination, present a major problem in biology. Sex typically involves recombination of closely related DNA or RNA sequences, which is fundamentally a random process that creates but also breaks up beneficial allele combinations. Directed evolution experiments, which combine in vitro mutation and recombination protocols with in vitro or in vivo selection, have proved to be an effective approach for improving functionality of nucleic acids and enzymes. As this approach allows extreme control over evolutionary conditions and parameters, it also facilitates the detection of small or position-specific recombination benefits and benefits associated with recombination between highly divergent genotypes. Yet, in vitro approaches have been largely exploratory and motivated by obtaining improved end products rather than testing hypotheses of recombination benefits. Here, we review the various experimental systems and approaches used by in vitro studies of recombination, discuss what they say about the evolutionary role of recombination, and sketch their potential for addressing extant questions about the evolutionary role of sex and recombination, in particular on complex fitness landscapes. We also review recent insights into the role of 'extracellular recombination' during the origin of life.This article is part of the themed issue 'Weird sex: the underappreciated diversity of sexual reproduction'. © 2016 The Author(s).

In vitro and in vivo approaches to study osteocyte biology.

Science.gov (United States)

Kalajzic, Ivo; Matthews, Brya G; Torreggiani, Elena; Harris, Marie A; Divieti Pajevic, Paola; Harris, Stephen E

2013-06-01

Osteocytes, the most abundant cell population of the bone lineage, have been a major focus in the bone research field in recent years. This population of cells that resides within mineralized matrix is now thought to be the mechanosensory cell in bone and plays major roles in the regulation of bone formation and resorption. Studies of osteocytes had been impaired by their location, resulting in numerous attempts to isolate primary osteocytes and to generate cell lines representative of the osteocytic phenotype. Progress has been achieved in recent years by utilizing in vivo genetic technology and generation of osteocyte directed transgenic and gene deficiency mouse models. We will provide an overview of the current in vitro and in vivo models utilized to study osteocyte biology. We discuss generation of osteocyte-like cell lines and isolation of primary osteocytes and summarize studies that have utilized these cellular models to understand the functional role of osteocytes. Approaches that attempt to selectively identify and isolate osteocytes using fluorescent protein reporters driven by regulatory elements of genes that are highly expressed in osteocytes will be discussed. In addition, recent in vivo studies utilizing overexpression or conditional deletion of various genes using dentin matrix protein (Dmp1) directed Cre recombinase are outlined. In conclusion, evaluation of the benefits and deficiencies of currently used cell lines/genetic models in understanding osteocyte biology underlines the current progress in this field. The future efforts will be directed towards developing novel in vitro and in vivo models that would additionally facilitate in understanding the multiple roles of osteocytes. Copyright © 2012 Elsevier Inc. All rights reserved.

Storage conditions of skin affect tissue structure and in vitro percutaneus penetration

DEFF Research Database (Denmark)

Nielsen, Jesper Bo; Bagatolli, Luis

2016-01-01

skin at -20oC causes structural changes in the upper Stratum Corneum observable with image techniques such as multiphoton excitation fluorescence microscopy. The presently available literature does, however, not support that the observed structural damage to the integrity is sufficient to cause...... structural changes in upper as well as deeper parts of Stratum Corneum. These more severe changes corresponds to significantly increased percutaneous penetration of chemicals applied to skin specimens stored at very low temperatures. Storage of human skin for later use in in vitro studies on percutaneous......For logistic and practical reasons it is difficult to perform in vitro studies on percutaneous penetration on fresh human skin obtained directly from surgery. Skin samples are therefore often kept frozen until use. The present chapter present the available literature on the topic. Storage of human...

TiF4 varnish protects the retention of brackets to enamel after in vitro mild erosive challenge.

Science.gov (United States)

Medeiros, Maria Isabel Dantas de; Carlo, Hugo Lemes; Santos, Rogério Lacerda Dos; Sousa, Frederico Barbosa; Castro, Ricardo Dias de; França, Renata Cristina Sobreira; Carvalho, Fabíola Galbiatti de

2018-05-14

The effect of fluoride agents on the retention of orthodontic brackets to enamel under erosive challenge is little investigated. The aim of this study was to evaluate the effect of titanium tetrafluoride (TiF4) and sodium fluoride (NaF) agents on the shear bond strength of brackets to enamel and on the enamel microhardness around brackets under erosive challenge. Brackets were bonded to bovine incisors. Five groups were formed according to fluoride application (n=10): TiF4 varnish, TiF4 solution, NaF varnish, NaF solution and control (without application). The specimens were submitted to erosive challenge (90 s cola drink/2h artificial saliva, 4x per day for 7 days). Solutions were applied before each erosive cycle and varnishes were applied once. Vickers Microhardness (VHN) was obtained before and after all cycles of erosion and the percentage of microhardness loss was calculated. Shear bond strength, adhesive remnant index and polarized light microscopy were conducted after erosion. The data were analyzed by ANOVA, Tukey, Kruskal-Wallis and Mann-Whitney U tests (α=0.05). The %VHN had no statistically significant differences among the experimental groups. However, considering the comparisons of all groups with the control group, TiF4 varnish showed the highest protection from enamel demineralization (effect size of 2.94, while the effect size for the other groups was >2.4). The TiF4 varnish group had significantly higher shear bond strength compared to other groups. There was no difference among groups for adhesive remnant index. Polarized light microscopy showed higher demineralization depth for the control group. Application of NaF and TiF4 agents during mild erosive challenge minimized the enamel mineral loss around brackets, however only the experimental TiF4 varnish was able to prevent the reduction of shear bond strength of brackets to enamel.

Short communication: measurements of methane emissions from feed samples in filter bags or dispersed in the medium in an in vitro gas production system.

Science.gov (United States)

Ramin, M; Krizsan, S J; Jančík, F; Huhtanen, P

2013-07-01

The objective of this study was to compare methane (CH4) emissions from different feeds when incubated within filter bags for in vitro analysis or directly dispersed in the medium in an automated gas in vitro system. Four different concentrates and 4 forages were used in this study. Two lactating Swedish Red cows were used for the collection of rumen fluid. Feed samples were milled to pass a 1.0-mm screen. Aliquots (0.5 g) of samples were weighed directly in the bottles or within the F 0285 filter bags that were placed in the bottles. Gas samples were taken during 24 and 48 h of incubation, and CH4 concentration was determined. The data were analyzed using a general linear model. Feeds differed significantly in CH4 emission both at 24 and at 48 h of incubation. The interaction between feed and method on methane emission in vitro was significant, indicating that the ranking of feeds was not consistent between the methods. Generally, greater amounts of CH4 were emitted from samples directly dispersed in the medium compared with those incubated within the filter bags, which could be a result of lower microbial activity within the filter bags. The ratio of CH4 to total gas was greater when the feeds were incubated within bags compared with samples directly dispersed in the medium. Incubating samples in filter bags during 48 h of incubation cannot be recommended for determination of CH4 emission of feeds in vitro. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

In vivo and in vitro toxicological effects of titanium dioxide nanoparticles on small intestine

Energy Technology Data Exchange (ETDEWEB)

Tassinari, Roberta; La Rocca, Cinzia; Tait, Sabrina; De Berardis, Barbara; Ammendolia, Maria Grazia; Iosi, Francesca; Di Virgilio, Antonio; Martinelli, Andrea; Maranghi, Francesca, E-mail: francesca.maranghi@iss.it [Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome (Italy); Stecca, Laura [European Commission-Joint Research Centre, Institute for Health and Consumer Protection Chemical assessment and Testing Unit, Via E. Fermi, 2749I-21027 Ispra (Italy)

2015-06-23

In European Union, titanium dioxide (TiO{sub 2}) as bulk material is a food additive (E171) and - as nanoparticle (NP) - is used as a white pigment in several products (e.g. food, cosmetics, drugs). E171 contains approximately 36% of particles less than 100 nm in at least one dimension and TiO{sub 2} NP exposure is estimated fairly below 2.5 mg/person/day. The gastrointestinal tract is a route of entry for NPs, thus representing a potential target of effects. In in vivo study, the effects of TiO{sub 2} NP in adult rat small intestine have been evaluated by oral administration of 0 (CTRL), 1 and 2 mg/kg body weight per day - relevant to human dietary intake. Detailed quali/quantitative histopathological analyses were performed on CTRL and treated rat samples. Scanning electron microscopy (SEM) analysis was performed on small intestine. An in vitro study on Caco-2 cells was also used in order to evaluate the potential cytotoxic effects directly on enterocytes through the lactate dehydrogenase (LDH) assay. Suspensions of TiO{sub 2} NPs for in vitro and in vivo study were characterized by EM. Histomorphometrical data showed treatment-related changes of villus height and widths in male rats. Significantly different from CTRL decreased LDH levels in the medium were detected in vitro at 24h with 2.5, 5, 10 and 20 µg/cm{sup 2} levels of TiO{sub 2} NPs. SEM analysis showed no damaged areas. Overall the results showed that enterocytes may represent a target of TiO{sub 2} NP toxicity by direct exposure both in vivo and in vitro models.

iNKT cell cytotoxic responses control T-lymphoma growth in vitro and in vivo

Science.gov (United States)

Bassiri, Hamid; Das, Rupali; Guan, Peng; Barrett, David M.; Brennan, Patrick J.; Banerjee, Pinaki P.; Wiener, Susan J.; Orange, Jordan S.; Brenner, Michael B.; Grupp, Stephan A.; Nichols, Kim E.

2013-01-01

Invariant natural killer T (iNKT) cells comprise a lineage of CD1d-restricted glycolipid-reactive T lymphocytes with important roles in host immunity to cancer. iNKT cells indirectly participate in antitumor responses by inducing dendritic cell maturation and producing cytokines that promote tumor clearance by CD8+ T and NK cells. Although iNKT cells thereby act as potent cellular adjuvants, it is less clear whether they directly control the growth of tumors. To gain insights into the direct contribution of iNKT cells to tumor immune surveillance, we developed in vitro and in vivo systems to selectively examine the antitumor activity of iNKT cells in the absence of other cytolytic effectors. Using the EL4 T-lymphoma cell line as a model, we find that iNKT cells exert robust and specific lysis of tumor cells in vitro in a manner that is differentially-induced by iNKT cell agonists of varying TCR affinities, such as OCH, α-galactosyl ceramide and PBS44. In vitro blockade of CD1d-mediated lipid antigen presentation, disruption of T cell receptor (TCR) signaling, or loss of perforin expression significantly reduce iNKT cell killing. Consistent with these findings, iNKT cell reconstitution of T, B, and NK cell-deficient mice slows EL4 growth in vivo via TCR-CD1d and perforin-dependent mechanisms. Together, these observations establish that iNKT cells are sufficient to control the growth of T-lymphoma in vitro and in vivo. They also suggest that the induction of iNKT cell cytotoxic responses in situ might serve as a more effective strategy to prevent and/or treat CD1d+ cancers, such as T-lymphoma. PMID:24563871

iNKT cell cytotoxic responses control T-lymphoma growth in vitro and in vivo .

Science.gov (United States)

Bassiri, Hamid; Das, Rupali; Guan, Peng; Barrett, David M; Brennan, Patrick J; Banerjee, Pinaki P; Wiener, Susan J; Orange, Jordan S; Brenner, Michael B; Grupp, Stephan A; Nichols, Kim E

2014-01-01

Invariant natural killer T (iNKT) cells comprise a lineage of CD1d-restricted glycolipid-reactive T lymphocytes with important roles in host immunity to cancer. iNKT cells indirectly participate in antitumor responses by inducing dendritic cell maturation and producing cytokines that promote tumor clearance by CD8+ T and NK cells. Although iNKT cells thereby act as potent cellular adjuvants, it is less clear whether they directly control the growth of tumors. To gain insights into the direct contribution of iNKT cells to tumor immune surveillance, we developed in vitro and in vivo systems to selectively examine the antitumor activity of iNKT cells in the absence of other cytolytic effectors. Using the EL4 T-lymphoma cell line as a model, we found that iNKT cells exert robust and specific lysis of tumor cells in vitro in a manner that is differentially induced by iNKT cell agonists of varying T-cell receptor (TCR) affinities, such as OCH, α-galactosyl ceramide, and PBS44. In vitro blockade of CD1d-mediated lipid antigen presentation, disruption of TCR signaling, or loss of perforin expression significantly reduce iNKT cell killing. Consistent with these findings, iNKT cell reconstitution of T, B, and NK cell–deficient mice slows EL4 growth in vivo via TCR-CD1d and perforin-dependent mechanisms. Together, these observations establish that iNKT cells are sufficient to control the growth of T lymphoma in vitro and in vivo. They also suggest that the induction of iNKT cell cytotoxic responses in situ might serve as a more effective strategy to prevent and/or treat CD1d+ cancers, such as T lymphoma. ©2013 AACR.

Clinical data management: Current status, challenges, and future directions from industry perspectives

Directory of Open Access Journals (Sweden)

Zhengwu Lu

2010-06-01

Full Text Available Zhengwu Lu1, Jing Su21Smith Hanley Consulting, Houston, Texas; 2Department of Chemical Engineering, University of Massachusetts, Amherst, MA, USAAbstract: To maintain a competitive position, the biopharmaceutical industry has been facing the challenge of increasing productivity both internally and externally. As the product of the clinical development process, clinical data are recognized to be the key corporate asset and provide critical evidence of a medicine’s efficacy and safety and of its potential economic value to the market. It is also well recognized that using effective technology-enabled methods to manage clinical data can enhance the speed with which the drug is developed and commercialized, hence enhancing the competitive advantage. The effective use of data-capture tools may ensure that high-quality data are available for early review and rapid decision-making. A well-designed, protocol-driven, standardized, site workflow-oriented and documented database, populated via efficient data feed mechanisms, will ensure regulatory and commercial questions receive rapid responses. When information from a sponsor’s clinical database or data warehouse develops into corporate knowledge, the value of the medicine can be realized. Moreover, regulators, payer groups, patients, activist groups, patient advocacy groups, and employers are becoming more educated consumers of medicine, requiring monetary value and quality, and seeking out up-todate medical information supplied by biopharmaceutical companies. All these developments in the current biopharmaceutical arena demand that clinical data management (CDM is at the forefront, leading change, influencing direction, and providing objective evidence. Sustaining an integrated database or data repository for initial product registration and subsequent postmarketing uses is a long-term process to maximize return on investment for organizations. CDM should be the owner of driving clinical data

Blood-filled cerebrospinal fluid-enhanced pericyte microvasculature contraction in rat retina: A novel in vitro study of subarachnoid hemorrhage

Science.gov (United States)

Liu, Zhi; Li, Qiang; Cui, Gaoyu; Zhu, Gang; Tang, Weihua; Zhao, Hengli; Zhang, John H.; Chen, Yujie; Feng, Hua

2016-01-01

Previously, it was widely accepted that the delayed ischemic injury and poor clinical outcome following subarachnoid hemorrhage (SAH) was caused by cerebral vasospasm. This classical theory was challenged by a clazosentan clinical trial, which failed to improve patient outcome, despite reversing angiographic vasospasm. One possible explanation for the results of this trial is the changes in microcirculation following SAH, particularly in pericytes, which are the primary cell type controlling microcirculation in the brain parenchyma. However, as a result of technical limitations and the lack of suitable models, there was no direct evidence of microvessel dysfunction following SAH. In the present study, whole-mount retinal microvasculature has been introduced to study microcirculation in the brain following experimental SAH in vitro. Artificial blood-filled cerebrospinal fluid (BSCF) was applied to the retinal microvasculature to test the hypothesis that the presence of subarachnoid blood affects the contractile properties of the pericytes containing cerebral microcirculation during the early phase of SAH. It was observed that BCSF induced retina microvessel contraction and that this contraction could be resolved by BCSF wash-out. Furthermore, BCSF application accelerated pericyte-populated collagen gel contraction and increased the expression of α-smooth muscle actin. In addition, BCSF induced an influx of calcium in cultured retinal pericytes. In conclusion, the present study demonstrates increased contractility of retinal microvessels and pericytes in the presence of BCSF in vitro. These findings suggest that pericyte contraction and microvascular dysfunction is induced following SAH, which could lead to greater susceptibility to SAH-induced ischemia. PMID:27698742

Testing in Vitro of an Apifitoterapeutic Formula Against Nosema spp.

OpenAIRE

Vasilica Savu; Ion Rădoi; Maria Magdici; Silvia Patruica; Nicoleta Ion; Agripina Sapcaliu

2015-01-01

Nosema, a parasitic disease that affects adult honey bees, has a directly correlation with the losses of bee colonies, until to depopulation. The target of our study was to determine the antinosema action of an apifitoterapeutic formula that was obtained in an earlier phase of our researches. In the present study, we have had two experiences (F and N) formed by clinically healthy bees. The experimental bees have received, in vitro, naturally infested honey (7 spores by Nosema spp / field). Th...

Parenting in Direct Provision: Parents' Perspectives Regarding Stresses and Supports

Science.gov (United States)

Ogbu, Helen Uchechukwu; Brady, Bernadine; Kinlen, Louise

2014-01-01

The Irish direct provision system for asylum seekers is acknowledged as providing a very challenging and exclusionary living environment for adults and children. To date, there has been little research focused specifically on the ways in which the direct provision environment impacts on the parenting role. This qualitative study explores the…

Comparison of Clobetasol Propionate Generics Using Simplified in Vitro Bioequivalence Method for Topical Drug Products.

Science.gov (United States)

Soares, Kelen Carine Costa; de Souza, Weidson Carlos; de Souza Texeira, Leonardo; da Cunha-Filho, Marcilio Sergio Soares; Gelfuso, Guilherme Martins; Gratieri, Tais

2017-11-20

The aim of this paper is to propose a simple in vitro skin penetration experiment in which the drug is extracted from the whole skin piece as a test valid for formulation screening and optimization during development process, equivalence assessment during quality control or post-approval after changes to the product. Twelve clobetasol propionate (CP) formulations (six creams and six ointments) from the local market were used as a model to challenge the proposed methodology in comparison to in vitro skin penetration following tape-stripping for drug extraction. To support the results, physicochemical tests for pH, viscosity, density and assay, as well as in vitro release were performed. Both protocols, extracting the drug from the skin using the tape-stripping technique or extracting from the full skin were capable of differentiating CP formulations. Only one formulation did not present statistical difference from the reference drug product in penetration tests and only other two oitments presented equivalent release to the reference. The proposed protocol is straightforward and reproducible. Results suggest the bioinequavalence of tested CP formulations reinforcing the necessity of such evaluations. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Ozone acts alone and synergistically with ionizing radiation to induce in vitro neoplastic transformation

Energy Technology Data Exchange (ETDEWEB)

Borek, C; Zaider, M; Ong, A; Mason, H; Witz, G

1986-09-01

Ozone, a major chemical oxidant in the atmosphere, is an environmental air pollutant whose ability to act as a direct carcinogen is unclear. Using in vitro transformation, a technique which permits the study of oncogenesis in the absence of host specific effects, it is reported for the first time that ozone (5 p.p.m. for 5 min) induces neoplastic transformation in vitro in both primary hamster embryo cells and mouse fibroblast cultures (C3H/10-1/2). Exposure of the hamster and mouse cells to ozone also results in enhanced levels of free radical-mediated lipid peroxidation products. The carcinogenic interaction between ozone and ionizing radiation is also reported. Exposure of the cells to 3 or 4 Gy of ..gamma..-rays, 2 h prior to O/sub 3/ treatment, results in markedly enhanced rates of transformation, statistically consistent with a synergistic interaction between the agents. The results demonstrate that O/sub 3/ acts as a direct carcinogen and co-carcinogen on susceptible cells, therefore having important consequences for public health.

The fitness landscape of HIV-1 gag: advanced modeling approaches and validation of model predictions by in vitro testing.

Directory of Open Access Journals (Sweden)

Jaclyn K Mann

2014-08-01

Full Text Available Viral immune evasion by sequence variation is a major hindrance to HIV-1 vaccine design. To address this challenge, our group has developed a computational model, rooted in physics, that aims to predict the fitness landscape of HIV-1 proteins in order to design vaccine immunogens that lead to impaired viral fitness, thus blocking viable escape routes. Here, we advance the computational models to address previous limitations, and directly test model predictions against in vitro fitness measurements of HIV-1 strains containing multiple Gag mutations. We incorporated regularization into the model fitting procedure to address finite sampling. Further, we developed a model that accounts for the specific identity of mutant amino acids (Potts model, generalizing our previous approach (Ising model that is unable to distinguish between different mutant amino acids. Gag mutation combinations (17 pairs, 1 triple and 25 single mutations within these predicted to be either harmful to HIV-1 viability or fitness-neutral were introduced into HIV-1 NL4-3 by site-directed mutagenesis and replication capacities of these mutants were assayed in vitro. The predicted and measured fitness of the corresponding mutants for the original Ising model (r = -0.74, p = 3.6×10-6 are strongly correlated, and this was further strengthened in the regularized Ising model (r = -0.83, p = 3.7×10-12. Performance of the Potts model (r = -0.73, p = 9.7×10-9 was similar to that of the Ising model, indicating that the binary approximation is sufficient for capturing fitness effects of common mutants at sites of low amino acid diversity. However, we show that the Potts model is expected to improve predictive power for more variable proteins. Overall, our results support the ability of the computational models to robustly predict the relative fitness of mutant viral strains, and indicate the potential value of this approach for understanding viral immune evasion

Protecting me from my Directive: Ensuring Appropriate Safeguards for Advance Directives in Dementia.

Science.gov (United States)

Auckland, Cressida

2018-02-01

With one in six people over 80 now suffering from dementia, advance directives provide an important means of empowerment. Upholding directives in the context of dementia, however, raises extra challenges, given the potential for the directive to conflict with an assessment of what is in the person's current best interests. Given the profound harm that tying a person with dementia to their previous wishes can do, it is essential that we have sufficient safeguards in place to ensure that we only uphold such directives where we can be sure they are truly autonomous and are intended to apply to the situation at hand-safeguards which are at present, severely lacking. This article will consider various mechanisms by which safeguards can be built into the legal regime to ensure that the original decision is autonomous, including making it mandatory for the person to undergo a consultation with a healthcare professional, which would involve a contemporaneous capacity assessment. Clinicians must also be confident that the directive applies to the situation at hand. Introducing formalities, including a standardised (though not mandatory) proforma, may help to enhance specificity about when the directive is triggered, and to what treatments it relates, to enable clinicians to better assess the directive's applicability. A national registry for advance directives might also be beneficial. It will be argued that health care professionals will have to play a much greater role in the drafting and registering of advance directives, if we are to feel comfortable in upholding them.

Sulfuric acid and hot water treatments enhance ex vitro and in vitro ...

African Journals Online (AJOL)

Seeds of Hibiscus dasycalyx S. F. Blake and Shiller, a federally listed candidate endangered species and native to North America and two variants of Hibiscus acetosella Welw. ex. Hiern were scarified using sulfuric acid and hot water. The effects of the scarification methods on in vitro and ex vitro germination in both ...

Recommendations for In Vitro and In Vivo Testing of Magnetic Nanoparticle Hyperthermia Combined with Radiation Therapy

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Spiridon V. Spirou

2018-05-01

Full Text Available Magnetic nanoparticle (MNP-mediated hyperthermia (MH coupled with radiation therapy (RT is a novel approach that has the potential to overcome various practical difficulties encountered in cancer treatment. In this work, we present recommendations for the in vitro and in vivo testing and application of the two treatment techniques. These recommendations were developed by the members of Working Group 3 of COST Action TD 1402: Multifunctional Nanoparticles for Magnetic Hyperthermia and Indirect Radiation Therapy (“Radiomag”. The purpose of the recommendations is not to provide definitive answers and directions but, rather, to outline those tests and considerations that a researcher must address in order to perform in vitro and in vivo studies. The recommendations are divided into 5 parts: (a in vitro evaluation of MNPs; (b in vitro evaluation of MNP-cell interactions; (c in vivo evaluation of the MNPs; (d MH combined with RT; and (e pharmacokinetic studies of MNPs. Synthesis and characterization of the MNPs, as well as RT protocols, are beyond the scope of this work.

Anti-Listeria starters: in vitro selection and production plant evaluation.

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Raimondi, Stefano; Popovic, Mina; Amaretti, Alberto; Di Gioia, Diana; Rossi, Maddalena

2014-05-01

Anti-Listeria bacterial starters are highly demanded by the meat industry. Novel bioprotective anti-Listeria starters were searched among Lactobacillus species strains isolated from artisanal sausages. The screening confirmed that anti-Listeria activity is a strain-specific property and yielded only 1 strain (of 36) exhibiting a satisfactory level of inhibition, L. delbrueckii WC0286. This strain was compared with two commercial bioprotective starters, SafePro B-SF-43 and SafePro B-LC-20, in a model simulating in vitro the first step of the fermentation process. The presence of the bioprotective starters did not modify the pH in such a way that could affect the safety or organoleptic properties of the product. Both SafePro B-SF-43 and SafePro B-LC-20 effected an important reduction of Listeria counts (0.56 and 0.72 log CFU g(-1), respectively, in 72 h), while the anti-Listeria effect of L. delbrueckii WC0286 was minor (0.15 log CFU g(-1)). These results discouraged the utilization of L. delbrueckii WC0286 for a challenge test in a pilot salami production, in favor of the best-performing bioprotective starter, SafePro B-LC-20. The test confirmed that SafePro B-LC-20 did not alter the acidification trend of sausages and was capable of inhibiting Listeria, which decreased by 1.21 log CFU g(-1). This information is relevant to address research activity toward the development of new bioprotective starters. The data herein presented demonstrate that the efficacy in Listeria control of potentially bioprotective bacterial starters requires further validation in real meat matrixes, possibly by using in vitro meat fermentation experiments to narrow down the list of candidates before pilot scale challenge tests.

Effectiveness of gamma-ray chronic irradiation on in vitro mutagenesis in crops

International Nuclear Information System (INIS)

Shigeki Nagatomi

2002-01-01

Effects of chronic or acute irradiations were compared using in vitro culture on inducing the mutation in model crops. In chrysanthemum, combined method with irradiation and in vitro culture can solve the problem of chimera formation in induced mutants, and provided 10 times greater mutation frequency than usual plant irradiation. The chronic culture method showed the widest color spectrum, whereas, the acute culture indicated a relatively low mutation rate and a very limited flower color spectrum in chrysanthemum. Flower color mutation of the regenerators could be induced more from petals and buds than from leaves. These facts are supposed that the gene loci fully expressed on floral organs may be unstable for mutation by mutagenesis or culture. It may be likely to control a direction of desired mutation on using explants with specific gene loci activated. In sugarcane, the chronic culture method extended quantitative characteristics of regenerated clonal lines toward not only the negative but positive direction. On the other hand, the acute culture method showed lower quantitative mutation as the irradiation dose rose. In chronic irradiation, regenerated mutant lines in sugarcane indicate generally little decrease in chromosome number and wider variations with relatively less damage. In acute irradiation, regenerated mutant lines show remarkable decrease of chromosome numbers in sugarcane mutant lines as the irradiation dose rose. There is close positive correlation between chromosome number and biomass of each mutant line. The chromosome number estimation is a proper indicator to monitor damage of adopted irradiation methods. Possible reason why the chronic culture methods indicate higher frequency and wider spectrum on mutation is demonstrated. . Problems solved and prospect of chronic irradiation and in vitro techniques are discussed. (Author)

Early death after feline infectious peritonitis virus challenge due to recombinant vaccinia virus immunization.

Science.gov (United States)

Vennema, H; de Groot, R J; Harbour, D A; Dalderup, M; Gruffydd-Jones, T; Horzinek, M C; Spaan, W J

1990-01-01

The gene encoding the fusogenic spike protein of the coronavirus causing feline infectious peritonitis was recombined into the genome of vaccinia virus. The recombinant induced spike-protein-specific, in vitro neutralizing antibodies in mice. When kittens were immunized with the recombinant, low titers of neutralizing antibodies were obtained. After challenge with feline infectious peritonitis virus, these animals succumbed earlier than did the control group immunized with wild-type vaccinia virus (early death syndrome). Images PMID:2154621

Brain Aggregates: An Effective In Vitro Cell Culture System Modeling Neurodegenerative Diseases.

Science.gov (United States)

Ahn, Misol; Kalume, Franck; Pitstick, Rose; Oehler, Abby; Carlson, George; DeArmond, Stephen J

2016-03-01

Drug discovery for neurodegenerative diseases is particularly challenging because of the discrepancies in drug effects between in vitro and in vivo studies. These discrepancies occur in part because current cell culture systems used for drug screening have many limitations. First, few cell culture systems accurately model human aging or neurodegenerative diseases. Second, drug efficacy may differ between dividing and stationary cells, the latter resembling nondividing neurons in the CNS. Brain aggregates (BrnAggs) derived from embryonic day 15 gestation mouse embryos may represent neuropathogenic processes in prion disease and reflect in vivo drug efficacy. Here, we report a new method for the production of BrnAggs suitable for drug screening and suggest that BrnAggs can model additional neurological diseases such as tauopathies. We also report a functional assay with BrnAggs by measuring electrophysiological activities. Our data suggest that BrnAggs could serve as an effective in vitro cell culture system for drug discovery for neurodegenerative diseases. © 2016 American Association of Neuropathologists, Inc. All rights reserved.

The challenges and possibilities of public access defibrillation

NARCIS (Netherlands)

Ringh, Mattias; Hollenberg, Jacob; Palsgaard-Moeller, Thea; Svensson, Leif; Rosenqvist, Mårten; Lippert, Freddy K.; Wissenberg, Mads; Malta Hansen, Carolina; Claesson, Andreas; Viereck, Soren; Zijlstra, Jolande A.; Koster, Rudolph W.; Herlitz, Johan; Blom, Marieke T.; Kramer-Johansen, Jo; Tan, Hanno L.; Beesems, Steffie; Hulleman, Michiel; Olasveengen, Theresa M.; Folke, Fredrik

2018-01-01

Out-of-hospital cardiac arrest (OHCA) is a major health problem that affects approximately four hundred and thousand patients annually in the United States alone. It is a major challenge for the emergency medical system as decreased survival rates are directly proportional to the time delay from

Bridging the Gap: Self-Directed Staff Technology Training

Directory of Open Access Journals (Sweden)

Kayla L. Quinney

2010-12-01

Full Text Available Undergraduates, as members of the Millennial Generation, are proficient in Web 2.0 technology and expect to apply these technologies to their coursework—including scholarly research. To remain relevant, academic libraries need to provide the technology that student patrons expect, and academic librarians need to learn and use these technologies themselves. Because leaders at the Harold B. Lee Library of Brigham Young University (HBLL perceived a gap in technology use between students and their staff and faculty, they developed and implemented the Technology Challenge, a self-directed technology training program that rewarded employees for exploring technology daily. The purpose of this paper is to examine the Technology Challenge through an analysis of results of surveys given to participants before and after the Technology Challenge was implemented. The program will also be evaluated in terms of the adult learning theories of andragogy and selfdirected learning. HBLL found that a self-directed approach fosters technology skills that librarians need to best serve students. In addition, it promotes lifelong learning habits to keep abreast of emerging technologies. This paper offers some insights and methods that could be applied in other libraries, the most valuable of which is the use of self-directed and andragogical training methods to help academic libraries better integrate modern technologies.

rAAV Vectors as Safe and Efficient Tools for the Stable Delivery of Genes to Primary Human Chondrosarcoma Cells In Vitro and In Situ

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Henning Madry

2012-01-01

Full Text Available Treatment of chondrosarcoma remains a major challenge in orthopaedic oncology. Gene transfer strategies based on recombinant adenoassociated viral (rAAV vectors may provide powerful tools to develop new, efficient therapeutic options against these tumors. In the present study, we tested the hypothesis that rAAV is adapted for a stable and safe delivery of foreign sequences in human chondrosarcoma tissue by transducing primary human chondrosarcoma cells in vitro and in situ with different reporter genes (E. coli lacZ, firefly luc, Discosoma sp. RFP. The effects of rAAV administration upon cell survival and metabolic activities were also evaluated to monitor possibly detrimental effects of the gene transfer method. Remarkably, we provide evidence that efficient and prolonged expression of transgene sequences via rAAV can be safely achieved in all the systems investigated, demonstrating the potential of the approach of direct application of therapeutic gene vectors as a means to treat chondrosarcoma.

Direct strip casting of steel - historical perspective and future direction

International Nuclear Information System (INIS)

Manohar, P.A.; Hunter, A.; Ferry, M.

2000-01-01

The commercialisation of direct strip casting (DSC) of steel represents the realisation of a dream cherished by engineers for over one hundred and fifty years. The story of the global competition for DSC of steel, that has ingredients of romanticism of chasing of a dream, adventure and intrigue, is being played out across continents over dozens of decades with an interplay of setbacks and successes. At this stage, DSC is set to make a profound impact on the steelmaking landscape. This paper reviews the important milestones in this compelling story, presents the current status and then gazes into the crystal ball in an attempt to predict which turn the story may take in the near future. The constraints and critical challenges for the successful commercialisation of DSC are highlighted. Recent results are discussed relating the production process to quality control and properties of unalloyed, low carbon strip-cast steels. future metallurgical challenges include a better understanding of solidification mechanism during high-speed casting and secondary processing variables affecting the final microstructure of austenitic grains

In Vitro Antidiabetic Effects and Antioxidant Potential of Cassia nemophila Pods

Directory of Open Access Journals (Sweden)

Gauhar Rehman

2018-01-01

Full Text Available The antidiabetic and antioxidant potential of ethanolic extract of Cassia nemophila pod (EECNP was evaluated by three in vitro assays, including yeast glucose uptake assay, glucose adsorption assay, and DPPH radical scavenging activity. The result revealed that the extracts have enhanced the uptake of glucose through the plasma membrane of yeast cells. A linear increase in glucose uptake by yeast cells was noticed with gradual increase in the concentration of the test samples. Moreover, the adsorption capacity of the EECNP was directly proportional to the molar concentration of glucose. Also, the DPPH radical scavenging capacity of the extract was increased to a maximum value of 43.3% at 80 μg/ml, which was then decreased to 41.9% at 100 μg/ml. From the results, it was concluded that EECNP possess good antidiabetic and antioxidant properties as shown by in vitro assays.

In vitro and in vivo antimicrobial activities of seeds of Caesalpinia bonduc (Lin.) Roxb.

Science.gov (United States)

Arif, Tasleem; Mandal, T K; Kumar, Naresh; Bhosale, J D; Hole, Archana; Sharma, G L; Padhi, M M; Lavekar, G S; Dabur, Rajesh

2009-05-04

Caesalpinia bonduc (Lin.) Roxb. is a known drug in Ayurveda to treat various diseases specifically tumors, cysts and cystic fibrosis (CF). The aim of this study was to assess in vitro as well as in vivo antimicrobial activity of Caesalpinia bonduc seeds. The in vitro antimicrobial activities of seed coat and seed kernel extracts were investigated by microbroth dilution assay. In vivo activities of hydro-alcoholic extracts were investigated in rat models of chronic Pseudomonas aeruginosa pneumonia mimicking that in patients with cystic fibrosis. Various extracts of plant seeds exhibited in vitro antimicrobial activities in a range of 22-350 microg/ml. The extracts also showed activity against methicillin resistant (MR) Staphylococcus aureus and ampicillin resistant (AR) Pseudomonas aeruginosa as in the sensitive strains. In rat model of chronic Pseudomonas aeruginosa pneumonia, hydro-alcoholic extracts of Caesalpinia bonduc seed kernel (CBSK) and Caesalpinia bonduc seed coat (CBSC) were injected subcutaneously in the test groups of animals. The control groups were treated with cortisone and saline. Two weeks after challenge with Pseudomonas aeruginosa, the CBSK treated animals showed a significant bacterial clearance from the lungs (PCaesalpinia bonduc may have the potential to be promising natural medicine, with other forms of treatments, for CF patients with chronic Pseudomonas aeruginosa lung infections.

Material properties and in vitro biocompatibility of a newly developed bone cement

Directory of Open Access Journals (Sweden)

Elke Mitzner

2009-01-01

Full Text Available In this study mechanical properties and biocompatibility (In Vitro of a new bone cement were investigated. A new platform technology named COOL is a variable composite of dissolved, chemically modified PMMA and different bioceramics. COOL cures at body temperature via a classical cementation reaction. Compressive strengths ranging from 3.6 ± 0.8 to 62.8 ± 1.3 MPa and bending strengths ranging from 9.9 ± 2.4 to 26.4 ± 3.0 MPa were achieved with different COOL formulations. Porosity varied between 31 and 43%. Varying the components of each formulation mechanical properties and porosity could be adjusted. In Vitro biocompatibility studies with primary human osteoblasts (pHOB in direct contact with different COOL formulations, did not reveal any signs of toxicity. In contrast to Refobacin® R, cells incubated with COOL showed similar density, viability and ALP activity compared to control, if specimen were added immediately to the cell monolayer after preparation. In conclusion, COOL has promising mechanical properties in combination with high biocompatibility In Vitro and combines different advantages of both CPCs and PMMA cements by avoiding some of the respective shortcomings.

Three-dimensional printing of Hela cells for cervical tumor model in vitro

International Nuclear Information System (INIS)

Zhao, Yu; Yao, Rui; Ouyang, Liliang; Ding, Hongxu; Zhang, Ting; Sun, Wei; Zhang, Kaitai; Cheng, Shujun

2014-01-01

Advances in three-dimensional (3D) printing have enabled the direct assembly of cells and extracellular matrix materials to form in vitro cellular models for 3D biology, the study of disease pathogenesis and new drug discovery. In this study, we report a method of 3D printing for Hela cells and gelatin/alginate/fibrinogen hydrogels to construct in vitro cervical tumor models. Cell proliferation, matrix metalloproteinase (MMP) protein expression and chemoresistance were measured in the printed 3D cervical tumor models and compared with conventional 2D planar culture models. Over 90% cell viability was observed using the defined printing process. Comparisons of 3D and 2D results revealed that Hela cells showed a higher proliferation rate in the printed 3D environment and tended to form cellular spheroids, but formed monolayer cell sheets in 2D culture. Hela cells in 3D printed models also showed higher MMP protein expression and higher chemoresistance than those in 2D culture. These new biological characteristics from the printed 3D tumor models in vitro as well as the novel 3D cell printing technology may help the evolution of 3D cancer study. (paper)

Boosters and barriers for direct cardiac reprogramming.

Science.gov (United States)

Talkhabi, Mahmood; Zonooz, Elmira Rezaei; Baharvand, Hossein

2017-06-01

Heart disease is currently the most significant cause of morbidity and mortality worldwide, which accounts for approximately 33% of all deaths. Recently, a promising and alchemy-like strategy has been developed called direct cardiac reprogramming, which directly converts somatic cells such as fibroblasts to cardiac lineage cells such as cardiomyocytes (CMs), termed induced CMs or iCMs. The first in vitro cardiac reprogramming study, mediated by cardiac transcription factors (TFs)-Gata4, Tbx5 and Mef2C-, was not enough efficient to produce an adequate number of fully reprogrammed, functional iCMs. As a result, numerous combinations of cardiac TFs exist for direct cardiac reprogramming of mouse and human fibroblasts. However, the efficiency of direct cardiac reprogramming remains low. Recently, a number of cellular and molecular mechanisms have been identified to increase the efficiency of direct cardiac reprogramming and the quality of iCMs. For example, microgrooved substrate, cardiogenic growth factors [VEGF, FGF, BMP4 and Activin A], and an appropriate stoichiometry of TFs boost the direct cardiac reprogramming. On the other hand, serum, TGFβ signaling, activators of epithelial to mesenchymal transition, and some epigenetic factors (Bmi1 and Ezh2) are barriers for direct cardiac reprogramming. Manipulating these mechanisms by the application of boosters and removing barriers can increase the efficiency of direct cardiac reprogramming and possibly make iCMs reliable for cell-based therapy or other potential applications. In this review, we summarize the latest trends in cardiac TF- or miRNA-based direct cardiac reprogramming and comprehensively discuses all molecular and cellular boosters and barriers affecting direct cardiac reprogramming. Copyright © 2017 Elsevier Inc. All rights reserved.

A new in vitro lipid digestion - in vivo absorption model to evaluate the mechanisms of drug absorption from lipid-based formulations.

Science.gov (United States)

Crum, Matthew F; Trevaskis, Natalie L; Williams, Hywel D; Pouton, Colin W; Porter, Christopher J H

2016-04-01

In vitro lipid digestion models are commonly used to screen lipid-based formulations (LBF), but in vitro-in vivo correlations are in some cases unsuccessful. Here we enhance the scope of the lipid digestion test by incorporating an absorption 'sink' into the experimental model. An in vitro model of lipid digestion was coupled directly to a single pass in situ intestinal perfusion experiment in an anaesthetised rat. The model allowed simultaneous real-time analysis of the digestion and absorption of LBFs of fenofibrate and was employed to evaluate the influence of formulation digestion, supersaturation and precipitation on drug absorption. Formulations containing higher quantities of co-solvent and surfactant resulted in higher supersaturation and more rapid drug precipitation in vitro when compared to those containing higher quantities of lipid. In contrast, when the same formulations were examined using the coupled in vitro lipid digestion - in vivo absorption model, drug flux into the mesenteric vein was similar regardless of in vitro formulation performance. For some drugs, simple in vitro lipid digestion models may underestimate the potential for absorption from LBFs. Consistent with recent in vivo studies, drug absorption for rapidly absorbed drugs such as fenofibrate may occur even when drug precipitation is apparent during in vitro digestion.

Biomedical engineering at UCT - challenges and opportunities.

Science.gov (United States)

Douglas, Tania S

2012-03-02

The biomedical engineering programme at the University of Cape Town has the potential to address some of South Africa's unique public health challenges and to contribute to growth of the local medical device industry, directly and indirectly, through research activities and postgraduate education. Full realisation of this potential requires engagement with the clinical practice environment and with industry.

An in vitro approach for comparative interspecies metabolism of agrochemicals.

Science.gov (United States)

Whalley, Paul M; Bartels, Michael; Bentley, Karin S; Corvaro, Marco; Funk, Dorothee; Himmelstein, Matthew W; Neumann, Birgit; Strupp, Christian; Zhang, Fagen; Mehta, Jyotigna

2017-08-01

The metabolism and elimination of a xenobiotic has a direct bearing on its potential to cause toxicity in an organism. The confidence with which data from safety studies can be extrapolated to humans depends, among other factors, upon knowing whether humans are systemically exposed to the same chemical entities (i.e. a parent compound and its metabolites) as the laboratory animals used to study toxicity. Ideally, to understand a metabolite in terms of safety, both the chemical structure and the systemic exposure would need to be determined. However, as systemic exposure data (i.e. blood concentration/time data of test material or metabolites) in humans will not be available for agrochemicals, an in vitro approach must be taken. This paper outlines an in vitro experimental approach for evaluating interspecies metabolic comparisons between humans and animal species used in safety studies. The aim is to ensure, where possible, that all potential human metabolites are also present in the species used in the safety studies. If a metabolite is only observed in human in vitro samples and is not present in a metabolic pathway defined in the toxicological species already, the toxicological relevance of this metabolite must be evaluated. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Activity-based in vitro selection of T4 DNA ligase

International Nuclear Information System (INIS)

Takahashi, Fumio; Funabashi, Hisakage; Mie, Masayasu; Endo, Yaeta; Sawasaki, Tatsuya; Aizawa, Masuo; Kobatake, Eiry

2005-01-01

Recent in vitro methodologies for selection and directed evolution of proteins have concentrated not only on proteins with affinity such as single-chain antibody but also on enzymes. We developed a display technology for selection of T4 DNA ligase on ribosome because an in vitro selection method for DNA ligase had never been developed. The 3' end of mRNA encoding the gene of active or inactive T4 DNA ligase-spacer peptide fusion protein was hybridized to dsDNA fragments with cohesive ends, the substrate of T4 DNA ligase. After in vitro translation of the mRNA-dsDNA complex in a rabbit reticulocyte system, a mRNA-dsDNA-ribosome-ligase complex was produced. T4 DNA ligase enzyme displayed on a ribosome, through addition of a spacer peptide, is able to react with dsDNA in the complex. The complex expressing active ligase was biotinylated by ligation with another biotinylated dsDNA probe and selected with streptavidin-coated magnetic beads. We effectively selected active T4 DNA ligase from a small amount of protein. The gene of the active T4 DNA ligase was enriched 40 times from a mixture of active and inactive genes using this selection strategy. This ribosomal display strategy may have high potential to be useful for selection of other enzymes associated with DNA

Irisin Enhances Osteoblast Differentiation In Vitro

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Graziana Colaianni

2014-01-01

Full Text Available It has been recently demonstrated that exercise activity increases the expression of the myokine Irisin in skeletal muscle, which is able to drive the transition of white to brown adipocytes, likely following a phenomenon of transdifferentiation. This new evidence supports the idea that muscle can be considered an endocrine organ, given its ability to target adipose tissue by promoting energy expenditure. In accordance with these new findings, we hypothesized that Irisin is directly involved in bone metabolism, demonstrating its ability to increase the differentiation of bone marrow stromal cells into mature osteoblasts. Firstly, we confirmed that myoblasts from mice subjected to 3 weeks of free wheel running increased Irisin expression compared to nonexercised state. The conditioned media (CM collected from myoblasts of exercised mice induced osteoblast differentiation in vitro to a greater extent than those of mice housed in resting conditions. Furthermore, the differentiated osteoblasts increased alkaline phosphatase and collagen I expression by an Irisin-dependent mechanism. Our results show, for the first time, that Irisin directly targets osteoblasts, enhancing their differentiation. This finding advances notable perspectives in future studies which could satisfy the ongoing research of exercise-mimetic therapies with anabolic action on the skeleton.

Non-viral delivery systems for CRISPR/Cas9-based genome editing: Challenges and opportunities.

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Li, Ling; Hu, Shuo; Chen, Xiaoyuan

2018-07-01

In recent years, CRISPR (clustered regularly interspaced short palindromic repeat)/Cas (CRISPR-associated) genome editing systems have become one of the most robust platforms in basic biomedical research and therapeutic applications. To date, efficient in vivo delivery of the CRISPR/Cas9 system to the targeted cells remains a challenge. Although viral vectors have been widely used in the delivery of the CRISPR/Cas9 system in vitro and in vivo, their fundamental shortcomings, such as the risk of carcinogenesis, limited insertion size, immune responses and difficulty in large-scale production, severely limit their further applications. Alternative non-viral delivery systems for CRISPR/Cas9 are urgently needed. With the rapid development of non-viral vectors, lipid- or polymer-based nanocarriers have shown great potential for CRISPR/Cas9 delivery. In this review, we analyze the pros and cons of delivering CRISPR/Cas9 systems in the form of plasmid, mRNA, or protein and then discuss the limitations and challenges of CRISPR/Cas9-based genome editing. Furthermore, current non-viral vectors that have been applied for CRISPR/Cas9 delivery in vitro and in vivo are outlined in details. Finally, critical obstacles for non-viral delivery of CRISPR/Cas9 system are highlighted and promising strategies to overcome these barriers are proposed. Published by Elsevier Ltd.

An in vitro model for screening estrogen activity of environmental samples after metabolism

Energy Technology Data Exchange (ETDEWEB)

Chahbane, N.; Schramm, K.W. [GSF - Forschungszentrum fuer Umwelt und Gesundheit Neuherberg GmbH, Oberschleissheim (Germany). Inst. fuer Oekologische Chemie; Kettrup, A. [Technische Univ. Muenchen, Freising (Germany). Lehrstuhl fuer Oekologische Chemie

2004-09-15

For a few years, yeast estrogen assay (YES) was accepted as a reliable and economic model for screening of environmental estrogens. Though the chemicals directly act with estrogen receptor (ER) can be filtered out by this model, there are still chemicals act with ER only after metabolism and some chemicals eliminate their estrogen activities after metabolism. That is to say, their metabolites exert or have stronger estrogen activities than themselves, which can be called bio-activation. In this case, for the lack of the metabolism enzyme system as human and other animals, only the assay with recombinant yeast cells is insufficient. So, it is necessary to combine the YES with metabolism procedure to evaluate the estrogen activities of these chemicals. The most common method used currently for in vitro metabolic activation in mutagenicity testing and also be applied to the estrogen screening field is S-9 mixture. Also, there is an attempt to develop a chemical model for cytochrome P450 as a bio-mimetic metabolic activation system. All these methods can be used as in vitro models for metabolism. Compare with these models, using whole H4II E cells for metabolism is an alternative and with superiorities. It has the excellence of short experiment period as all other in vitro models, but is much more close to the real surroundings as in vivo. Furthermore, the activity of 7-ethoxyresorufin-O-deethylase (EROD) can be easily measured during the whole incubation period for us to discuss the metabolic activities in a quantitative foundation, not only in qualitative. Methoxychlor is one of the chemicals with bio-activation ability. When directly used in the YES, it shows weak estrogen activity. But a main metabolite of methoxychlor, 2,2-bis (p-hydroxyphenyl) - 1,1,1-trichloroethane (HPTE) is a known estrogen mimic. For the long time using methoxychlor as a pesticide and its clear background, it is an ideal chemical to establish this in vitro system.

Evaluating Foreign Direct Investment and Africa's Development ...

African Journals Online (AJOL)

The question of Africa's development has continued to occupy the front burner from the social and economic discussions by scholars of various divides. But Africa's development through foreign direct investment has become a recent challenge to the African continent. African social critics and commentators as well as ...

Through the smoke: Use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility

International Nuclear Information System (INIS)

Camlin, Nicole J.; McLaughlin, Eileen A.; Holt, Janet E.

2014-01-01

A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents’ in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes. - Highlights: • In vivo exposure to individual cigarette smoke chemicals alters female fertility. • The use of in vitro models in determining molecular mechanisms • Whole cigarette smoke inhalation animal models negatively affect ovarian function

Through the smoke: Use of in vivo and in vitro cigarette smoking models to elucidate its effect on female fertility

Energy Technology Data Exchange (ETDEWEB)

Camlin, Nicole J. [School of Environment and Life Sciences, University of Newcastle, Callaghan, NSW 2308 (Australia); McLaughlin, Eileen A., E-mail: eileen.mclaughlin@newcastle.edu.au [School of Environment and Life Sciences, University of Newcastle, Callaghan, NSW 2308 (Australia); Holt, Janet E. [School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW 2308 (Australia)

2014-12-15

A finite number of oocytes are established within the mammalian ovary prior to birth to form a precious ovarian reserve. Damage to this limited pool of gametes by environmental factors such as cigarette smoke and its constituents therefore represents a significant risk to a woman's reproductive capacity. Although evidence from human studies to date implicates a detrimental effect of cigarette smoking on female fertility, these retrospective studies are limited and present conflicting results. In an effort to more clearly understand the effect of cigarette smoke, and its chemical constituents, on female fertility, a variety of in vivo and in vitro animal models have been developed. This article represents a systematic review of the literature regarding four of experimental model types: 1) direct exposure of ovarian cells and follicles to smoking constituents’ in vitro, 2) direct exposure of whole ovarian tissue with smoking constituents in vitro, 3) whole body exposure of animals to smoking constituents and 4) whole body exposure of animals to cigarette smoke. We summarise key findings and highlight the strengths and weaknesses of each model system, and link these to the molecular mechanisms identified in smoke-induced fertility changes. - Highlights: • In vivo exposure to individual cigarette smoke chemicals alters female fertility. • The use of in vitro models in determining molecular mechanisms • Whole cigarette smoke inhalation animal models negatively affect ovarian function.

Benfotiamine exhibits direct antioxidative capacity and prevents induction of DNA damage in vitro.

Science.gov (United States)

Schmid, Ursula; Stopper, Helga; Heidland, August; Schupp, Nicole

2008-01-01

Complications in diabetes mellitus are partially mediated by enhanced formation of reactive oxygen species. Among the factors involved in reactive oxygen species formation, advanced glycation end products play a key role. Owing to a reduced activity of the enzyme transketolase, which requires diphosphorylated thiamine (vitamin B(1)) as cofactor, an accumulation of those deleterious glucose metabolites especially in diabetic patients can be observed. Benfotiamine, a lipophilic thiamine diphosphate prodrug, prevented early renal and retinal changes in animal studies, and reduced neuropathic pain in clinical studies. Several mechanisms for these activities have been described. We investigated for the first time direct antioxidant abilities of benfotiamine. Additionally, a potential DNA protective effect of benfotiamine was analysed. Oxidative stress was detected by flow cytometry, antioxidative capacity was measured with the ferric reducing ability of plasma (FRAP) assay, two endpoints for genomic damage were assessed: the comet assay and the micronucleus test, and the expression and activity of transketolase was quantified. Benfotiamine prevented oxidative stress induced by the mutagen 4-nitroquinoline-1-oxide (NQO), the uremic toxin indoxyl sulfate, and the peptide hormone angiotensin II in three different kidney cell lines. Cell-free experiments showed a direct antioxidant effect of benfotiamine, which might account for the protective effect. Oxidative DNA damage, induced by angiotensin II, was completely prevented by benfotiamine. Incubation with benfotiamine increased transketolase expression and activity in the cells. Benfotiamine shows a direct antioxidant action. This effect of benfotiamine may be involved in the improvement of diabetic late complications, including peripheral neuropathy.

From Chemotherapy to Combined Targeted Therapeutics: In Vitro and in Vivo Models to Decipher Intra-tumor Heterogeneity

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Guido Gambara

2018-02-01

Full Text Available Recent advances in next-generation sequencing and other omics technologies capable to map cell fate provide increasing evidence on the crucial role of intra-tumor heterogeneity (ITH for cancer progression. The different facets of ITH, from genomic to microenvironmental heterogeneity and the hierarchical cellular architecture originating from the cancer stem cell compartment, contribute to the range of tumor phenotypes. Decoding these complex data resulting from the analysis of tumor tissue complexity poses a challenge for developing novel therapeutic strategies that can counteract tumor evolution and cellular plasticity. To achieve this aim, the development of in vitro and in vivo cancer models that resemble the complexity of ITH is crucial in understanding the interplay of cells and their (microenvironment and, consequently, in testing the efficacy of new targeted treatments and novel strategies of tailoring combinations of treatments to the individual composition of the tumor. This challenging approach may be an important cornerstone in overcoming the development of pharmaco-resistances during multiple lines of treatment. In this paper, we report the latest advances in patient-derived 3D (PD3D cell cultures and patient-derived tumor xenografts (PDX as in vitro and in vivo models that can retain the genetic and phenotypic heterogeneity of the tumor tissue.

Outils d’évaluation in vitro de la capacité antioxydante

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Laguerre Mickaël

2007-09-01

Full Text Available Oxidative degradation of lipids leads to quality deterioration of foods and cosmetics and has received increasing attention as a causative or associative factor in a variety of pathological conditions. A current very promising way to counteract lipid oxidation is to use vegetable antioxidants for nutritional, therapeutic or food quality preservation purpose. A major challenge is to develop analytical tools to assess the real antioxidant power of these molecules. In this way, many in vitro tests are now available, which monitor generally the inhibitory effect of antioxidant on the oxidative degradation of lipid or lipid-like substrate under natural or artificial oxidizing conditions. Overall, this measurement necessitates focusing either on the oxidizable substrate loss, or on the formation of oxidation products. To perform the first strategy, fluorophores such as cis-parinaric acid, BODIPY, and phospholipidic derivatives of fluorescein are often used, whereas the second employs usually linoleic acid, its methyl ester or natural lipids. After a brief recall of antioxidation mechanisms, this review presents a panorama of the available in vitro methodologies and of the last advances in this field. A comparative study between methods will be also discussed in terms of advantages and drawbacks.

Five Decades: From Challenge to Acclaim.

Science.gov (United States)

Melina, Vesanto

2016-09-01

What can make your work as a dietitian so meaningful that you begin each day with enthusiasm, and if you so choose, retain that joy in your work for 5 decades or more? Three themes are: (i) doing work that profoundly makes sense to you, (ii) inspiring others (and yourself) to make healthful choices, and (iii) moving through challenges to success. Initially it can be challenging to make a living through work that is most deeply meaningful or closest to your heart. Yet it is well worth finding the balance between practicality and movement in the desired direction. Other challenges faced by dietitians involve helping others to adopt new, more healthful lifestyle choices. As health professionals, our attitudes towards plant-based diets have changed dramatically during these past decades. This article examines our evolving perspectives of plant-based diets, and uses this as an example of movement through challenges to success and acclaim. Vegetarian and vegan diets that were considered entirely inappropriate for many stages of the life cycle in the 1970s are now seen to confer health benefits. This applies to well-designed plant-based diets, thus offering a significant role for dietitians as creative leaders in this field.

Spontaneous transformation of adult mesenchymal stem cells from cynomolgus macaques in vitro

International Nuclear Information System (INIS)

Ren, Zhenhua; Wang, Jiayin; Zhu, Wanwan; Guan, Yunqian; Zou, Chunlin; Chen, Zhiguo; Zhang, Y. Alex

2011-01-01

Mesenchymal stem cells (MSCs) have shown potential clinical utility in cell therapy and tissue engineering, due to their ability to proliferate as well as to differentiate into multiple lineages, including osteogenic, adipogenic, and chondrogenic specifications. Therefore, it is crucial to assess the safety of MSCs while extensive expansion ex vivo is a prerequisite to obtain the cell numbers for cell transplantation. Here we show that MSCs derived from adult cynomolgus monkey can undergo spontaneous transformation following in vitro culture. In comparison with MSCs, the spontaneously transformed mesenchymal cells (TMCs) display significantly different growth pattern and morphology, reminiscent of the characteristics of tumor cells. Importantly, TMCs are highly tumorigenic, causing subcutaneous tumors when injected into NOD/SCID mice. Moreover, no multiple differentiation potential of TMCs is observed in vitro or in vivo, suggesting that spontaneously transformed adult stem cells may not necessarily turn into cancer stem cells. These data indicate a direct transformation of cynomolgus monkey MSCs into tumor cells following long-term expansion in vitro. The spontaneous transformation of the cultured cynomolgus monkey MSCs may have important implications for ongoing clinical trials and for models of oncogenesis, thus warranting a more strict assessment of MSCs prior to cell therapy. -- Highlights: ► Spontaneous transformation of cynomolgus monkey MSCs in vitro. ► Transformed mesenchymal cells lack multipotency. ► Transformed mesenchymal cells are highly tumorigenic. ► Transformed mesenchymal cells do not have the characteristics of cancer stem cells.

ISDD: A computational model of particle sedimentation, diffusion and target cell dosimetry for in vitro toxicity studies

Science.gov (United States)

2010-01-01

Background The difficulty of directly measuring cellular dose is a significant obstacle to application of target tissue dosimetry for nanoparticle and microparticle toxicity assessment, particularly for in vitro systems. As a consequence, the target tissue paradigm for dosimetry and hazard assessment of nanoparticles has largely been ignored in favor of using metrics of exposure (e.g. μg particle/mL culture medium, particle surface area/mL, particle number/mL). We have developed a computational model of solution particokinetics (sedimentation, diffusion) and dosimetry for non-interacting spherical particles and their agglomerates in monolayer cell culture systems. Particle transport to cells is calculated by simultaneous solution of Stokes Law (sedimentation) and the Stokes-Einstein equation (diffusion). Results The In vitro Sedimentation, Diffusion and Dosimetry model (ISDD) was tested against measured transport rates or cellular doses for multiple sizes of polystyrene spheres (20-1100 nm), 35 nm amorphous silica, and large agglomerates of 30 nm iron oxide particles. Overall, without adjusting any parameters, model predicted cellular doses were in close agreement with the experimental data, differing from as little as 5% to as much as three-fold, but in most cases approximately two-fold, within the limits of the accuracy of the measurement systems. Applying the model, we generalize the effects of particle size, particle density, agglomeration state and agglomerate characteristics on target cell dosimetry in vitro. Conclusions Our results confirm our hypothesis that for liquid-based in vitro systems, the dose-rates and target cell doses for all particles are not equal; they can vary significantly, in direct contrast to the assumption of dose-equivalency implicit in the use of mass-based media concentrations as metrics of exposure for dose-response assessment. The difference between equivalent nominal media concentration exposures on a μg/mL basis and target cell

ISDD: A computational model of particle sedimentation, diffusion and target cell dosimetry for in vitro toxicity studies

Directory of Open Access Journals (Sweden)

Chrisler William B

2010-11-01

Full Text Available Abstract Background The difficulty of directly measuring cellular dose is a significant obstacle to application of target tissue dosimetry for nanoparticle and microparticle toxicity assessment, particularly for in vitro systems. As a consequence, the target tissue paradigm for dosimetry and hazard assessment of nanoparticles has largely been ignored in favor of using metrics of exposure (e.g. μg particle/mL culture medium, particle surface area/mL, particle number/mL. We have developed a computational model of solution particokinetics (sedimentation, diffusion and dosimetry for non-interacting spherical particles and their agglomerates in monolayer cell culture systems. Particle transport to cells is calculated by simultaneous solution of Stokes Law (sedimentation and the Stokes-Einstein equation (diffusion. Results The In vitro Sedimentation, Diffusion and Dosimetry model (ISDD was tested against measured transport rates or cellular doses for multiple sizes of polystyrene spheres (20-1100 nm, 35 nm amorphous silica, and large agglomerates of 30 nm iron oxide particles. Overall, without adjusting any parameters, model predicted cellular doses were in close agreement with the experimental data, differing from as little as 5% to as much as three-fold, but in most cases approximately two-fold, within the limits of the accuracy of the measurement systems. Applying the model, we generalize the effects of particle size, particle density, agglomeration state and agglomerate characteristics on target cell dosimetry in vitro. Conclusions Our results confirm our hypothesis that for liquid-based in vitro systems, the dose-rates and target cell doses for all particles are not equal; they can vary significantly, in direct contrast to the assumption of dose-equivalency implicit in the use of mass-based media concentrations as metrics of exposure for dose-response assessment. The difference between equivalent nominal media concentration exposures on a �

Sacarose e período de cultivo in vitro na aclimatização ex vitro de ginseng brasileiro (Pfaffia glomerata Spreng. Pedersen Sucrose and duration of in vitro growth on ex vitro acclimatization of Brazilian ginseng (Pfaffia glomerata Spreng. Pedersen

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Etiane Caldeira Skrebsky

2004-10-01

Full Text Available Pfaffia glomerata (Spreng. Pedersen é uma planta extensivamente usada na medicina popular em decorrência de possuir propriedades fitoterápicas. Devido à sua baixa capacidade fotossintética, as plantas cultivadas in vitro requerem uma fonte extra de carboidratos para suprir suas necessidades metabólicas. O tempo de cultivo in vitro influencia as taxas de crescimento das raízes e da parte aérea de P. glomerata. Este trabalho teve como objetivos avaliar os efeitos da sacarose e do período de cultivo in vitro na aclimatização ex vitro de plântulas de P. glomerata. Os tratamentos consistiram de uma combinação bifatorial (5x2 entre cinco concentrações de sacarose (15, 30, 45, 60 e 75g L-1 e dois períodos de cultivo in vitro (25 e 32 dias após a inoculação. No cultivo in vitro, a parcela experimental consistiu de um tubo de ensaio contendo 10mL de meio MS e um segmento nodal, obtidos de plântulas mantidas in vitro, de 1,0cm de comprimento e sem folhas. O procedimento de aclimatização consistiu de quatro fases sucessivas de cultivo ex vitro: (i abertura dos tubos de ensaio e exposição das plântulas ao ambiente de câmara climatizada por três dias; (ii transplantio para substrato Plantmax e cultivo em condições de câmara climatizada por 21 dias; (iii transferência das mudas para ambiente natural parcialmente sombreado, com duração de 24 dias; e (iv transplantio para solo em condições de cultivo a campo. O maior crescimento das plantas obtido pelo aumento da disponibilidade de sacarose (concentrações entre 45 e 60g L-1 no cultivo in vitro contribuiu para a aclimatização. Independente do período de retirada das plantas do cultivo in vitro, as mudas obtiveram adequada aclimatização. O procedimento de aclimatização foi 100% eficiente na produção de mudas de P. glomerata.Pfaffia glomerata (Spreng. Pedersen is an extensively used plant in popular medicine due to its phytotherapic characteristics. Due to low

Physiology of in vitro culture

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Maria Jesús Cañal

2001-01-01

Full Text Available The culture procedures described up to the eighties, did not made any mention to the environmental control of in vitro plant development. However, growth rate, development and many of the physiologic-morphologic features of the in vitro grown plants are influenced by the culture vessel. The increasing knowledge about the environmental control of culture vessels under sterile conditions, is helping to change micorpropagation procedures. The in vitro environment with lower rate ventilation, brings about low flow rates of matter and energy, with minimum variations of temperature, high relative humidity and large daily changes of the concentration of CO2 inside the culture vessel. The type of culture vessel (size, shape, fabric and closing system can influence the evolution of the atmosphere along the time of culture. Although submitted to different stresses factors plant can be grown in vitro, but plants can be faulty in their anatomy, morphology and physiology. As a consequence, these plants shown a phenotype unable to survive to ex vitro conditions. Different strategies can be used to control the atmosphere along the different phases of micropropagation, in heterotrophic, mixotrophic or autotrophic cultures. The election of the best strategy will be based on different factors as species, number of transplantes required, or quality-price relationship. enviromental control, tissue culture, micropropagation Keywords: in vitro enviromental, characteristic physiology,

Induction of mucosal immune responses and protection of cattle against direct-contact challenge by intranasal delivery with foot-and-mouth disease virus antigen mediated by nanoparticles

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Pan L

2014-12-01

Full Text Available Li Pan,1,2 Zhongwang Zhang,1,2 Jianliang Lv,1,2 Peng Zhou,1,2 Wenfa Hu,1,2 Yuzhen Fang,1,2 Haotai Chen,1,2 Xinsheng Liu,1,2 Junjun Shao,1,2 Furong Zhao,1,2 Yaozhong Ding,1,2 Tong Lin,1,2 Huiyun Chang,1,2 Jie Zhang,1,2 Yongguang Zhang,1,2 Yonglu Wang1,2 1State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS, Lanzhou, Gansu, People’s Republic of China; 2Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, People’s Republic of China Abstract: The aim of this study was to enhance specific mucosal, systemic, and cell-mediated immunity and to induce earlier onset of protection against direct-contact challenge in cattle by intranasal delivery of a nanoparticle-based nasal vaccine against type A foot-and-mouth disease (FMD. In this study, two kinds of nanoparticle-based nasal vaccines against type A FMD were designed: (1 chitosan-coated poly(lactic-co-glycolic acid (PLGA loaded with plasmid DNA (Chi-PLGA-DNA and (2 chitosan-trehalose and inactivated foot-and-mouth disease virus (FMDV (Chi-Tre-Inactivated. Cattle were immunized by an intranasal route with nanoparticles and then challenged for 48 hours by direct contact with two infected donor cattle per pen. Donors were inoculated intradermally in the tongue 48 hours before challenge, with 0.2 mL cattle-passaged FMDV. Serological and mucosal antibody responses were evaluated, and virus excretion and the number of contact infections were quantified. FMDV-specific secretory immunoglobulin (IgA (sIgA antibodies in nasal washes were initially detected at 4 days postvaccination (dpv with two kinds of nanoparticles. The highest levels of sIgA expression were observed in nasal washes, at 10 dpv, from animals with Chi-PLGA-DNA nanoparticles, followed by animals immunized once by intranasal route with

Directed evolution combined with synthetic biology strategies expedite semi-rational engineering of genes and genomes.

Science.gov (United States)

Kang, Zhen; Zhang, Junli; Jin, Peng; Yang, Sen

2015-01-01

Owing to our limited understanding of the relationship between sequence and function and the interaction between intracellular pathways and regulatory systems, the rational design of enzyme-coding genes and de novo assembly of a brand-new artificial genome for a desired functionality or phenotype are difficult to achieve. As an alternative approach, directed evolution has been widely used to engineer genomes and enzyme-coding genes. In particular, significant developments toward DNA synthesis, DNA assembly (in vitro or in vivo), recombination-mediated genetic engineering, and high-throughput screening techniques in the field of synthetic biology have been matured and widely adopted, enabling rapid semi-rational genome engineering to generate variants with desired properties. In this commentary, these novel tools and their corresponding applications in the directed evolution of genomes and enzymes are discussed. Moreover, the strategies for genome engineering and rapid in vitro enzyme evolution are also proposed.

Chaos: Challenges from and to socio-spatial form and policy

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Dimitrios S. Dendrinos

1997-01-01

Full Text Available A brief assessment is given of the major accomplishments made through the mathematics of chaos to the understanding of socio-spatial dynamics to date. Certain shortfalls are also presented, mostly associated with model testing and falsifiability which transcend socio-spatial dynamics. Beyond such shortcomings, lie an array of challenges for chaotic dynamics involving specifically socio-spatial form and policy. A few directions on meeting these challenges are suggested including the case of limited chaos.

Preparation & in vitro evaluation of 90Y-DOTA-rituximab

Science.gov (United States)

Kameswaran, Mythili; Pandey, Usha; Dash, Ashutosh; Samuel, Grace; Venkatesh, Meera

2016-01-01

Background & objectives: Radioimmunotherapy is extensively being used for the treatment of non-Hodgkin's lymphoma (NHL). Use of rituximab, a chimeric anti-CD20 antibody directed against the CD20 antigen in combination with suitable beta emitters is expected to result in good treatment response by its cross-fire and bystander effects. The present work involves the conjugation of p-isothiocyanatobenzyl DOTA (p-SCN-Bn-DOTA) to rituximab, its radiolabelling with 90Y and in vitro and in vivo evaluation to determine its potential as a radioimmunotherapeutic agent. Methods: Rituximab was conjugated with p-SCN-Bn-DOTA at 1:1 antibody: DOTA molar ratio. The number of DOTA molecules linked to one molecule of rituximab was determined by radioassay and spectroscopic assay. Radiolabelling of rituximab with 90Y was carried out and its in vitro stability was evaluated. In vitro cell binding studies were carried out in Raji cells expressing CD20 antigen. Biodistribution studies were carried out in normal Swiss mice. Results: Using both radioassay and spectroscopic method, it was determined that about five molecules of DOTA were linked to rituximab. Radiolabelling of the rituximab conjugate with 90Y and subsequent purification on PD-10 column gave a product with radiochemical purity (RCP) > 98 per cent which was retained at > 90 per cent up to 72 h when stored at 37°C. In vitro cell binding experiments of 90Y-DOTA-rituximab with Raji cells exhibited specific binding of 20.7 ± 0.1 per cent with 90Y-DOTA-rituximab which reduced to 15.5 ± 0.2 per cent when incubated with cold rituximab. The equilibrium constant Kd for 90Y-DOTA-Rituximab was determined to be 3.38 nM. Radiolabelled antibody showed clearance via hepatobiliary and renal routes and activity in tibia was found to be quite low indicating in vivo stability of 90Y-DOTA-rituximab. Interpretation & conclusions: p-SCN-Bn-DOTA was conjugated with rituximab and radiolabelling with 90Y was carried out. In vitro studies carried

In vitro gastrointestinal digestion increases the translocation of polystyrene nanoparticles in an in vitro intestinal co-culture model.

Science.gov (United States)

Walczak, Agata P; Kramer, Evelien; Hendriksen, Peter J M; Helsdingen, Richard; van der Zande, Meike; Rietjens, Ivonne M C M; Bouwmeester, Hans

2015-01-01

The conditions of the gastrointestinal tract may change the physicochemical properties of nanoparticles (NPs) and therewith the bioavailability of orally taken NPs. Therefore, we assessed the impact of in vitro gastrointestinal digestion on the protein corona of polystyrene NPs (PS-NPs) and their subsequent translocation across an in vitro intestinal barrier. A co-culture of intestinal Caco-2 and HT29-MTX cells was exposed to 50 nm PS-NPs of different charges (positive and negative) in two forms: pristine and digested in an in vitro gastrointestinal digestion model. In vitro digestion significantly increased the translocation of all, except the "neutral", PS-NPs. Upon in vitro digestion, translocation was 4-fold higher for positively charged NPs and 80- and 1.7-fold higher for two types of negatively charged NPs. Digestion significantly reduced the amount of protein in the corona of three out of four types of NPs. This reduction of proteins was 4.8-fold for "neutral", 3.5-fold for positively charged and 1.8-fold for one type of negatively charged PS-NPs. In vitro digestion also affected the composition of the protein corona of PS-NPs by decreasing the presence of higher molecular weight proteins and shifting the protein content of the corona to low molecular weight proteins. These findings are the first to report that in vitro gastrointestinal digestion significantly affects the protein corona and significantly increases the in vitro translocation of differently charged PS-NPs. These findings stress the importance of including the in vitro digestion in future in vitro intestinal translocation screening studies for risk assessment of orally taken NPs.

Applying evolutionary biology to address global challenges

Science.gov (United States)

Carroll, Scott P.; Jørgensen, Peter Søgaard; Kinnison, Michael T.; Bergstrom, Carl T.; Denison, R. Ford; Gluckman, Peter; Smith, Thomas B.; Strauss, Sharon Y.; Tabashnik, Bruce E.

2014-01-01

Two categories of evolutionary challenges result from escalating human impacts on the planet. The first arises from cancers, pathogens and pests that evolve too quickly, and the second from the inability of many valued species to adapt quickly enough. Applied evolutionary biology provides a suite of strategies to address these global challenges that threaten human health, food security, and biodiversity. This review highlights both progress and gaps in genetic, developmental and environmental manipulations across the life sciences that either target the rate and direction of evolution, or reduce the mismatch between organisms and human-altered environments. Increased development and application of these underused tools will be vital in meeting current and future targets for sustainable development. PMID:25213376

A Self-Folding Hydrogel In Vitro Model for Ductal Carcinoma.

Science.gov (United States)

Kwag, Hye Rin; Serbo, Janna V; Korangath, Preethi; Sukumar, Saraswati; Romer, Lewis H; Gracias, David H

2016-04-01

A significant challenge in oncology is the need to develop in vitro models that accurately mimic the complex microenvironment within and around normal and diseased tissues. Here, we describe a self-folding approach to create curved hydrogel microstructures that more accurately mimic the geometry of ducts and acini within the mammary glands, as compared to existing three-dimensional block-like models or flat dishes. The microstructures are composed of photopatterned bilayers of poly (ethylene glycol) diacrylate (PEGDA), a hydrogel widely used in tissue engineering. The PEGDA bilayers of dissimilar molecular weights spontaneously curve when released from the underlying substrate due to differential swelling ratios. The photopatterns can be altered via AutoCAD-designed photomasks so that a variety of ductal and acinar mimetic structures can be mass-produced. In addition, by co-polymerizing methacrylated gelatin (methagel) with PEGDA, microstructures with increased cell adherence are synthesized. Biocompatibility and versatility of our approach is highlighted by culturing either SUM159 cells, which were seeded postfabrication, or MDA-MB-231 cells, which were encapsulated in hydrogels; cell viability is verified over 9 and 15 days, respectively. We believe that self-folding processes and associated tubular, curved, and folded constructs like the ones demonstrated here can facilitate the design of more accurate in vitro models for investigating ductal carcinoma.

Validation of artificial skin equivalents as in vitro testing systems

Science.gov (United States)

Schmitt, Robert; Marx, Ulrich; Walles, Heike; Schober, Lena

2011-03-01

With the increasing complexity of the chemical composition of pharmaceuticals, cosmetics and everyday substances, the awareness of potential health issues and long term damages for humanoid organs is shifting into focus. Artificial in vitro testing systems play an important role in providing reliable test conditions and replacing precarious animal testing. Especially artificial skin equivalents ASEs are used for a broad spectrum of studies like penetration, irritation and corrosion of substances. One major challenge in tissue engineering is the qualification of each individual ASE as in vitro testing system. Due to biological fluctuations, the stratum corneum hornified layer of some ASEs may not fully develop or other defects might occur. For monitoring these effects we developed an fully automated Optical Coherence Tomography device. Here, we present different methods to characterize and evaluate the quality of the ASEs based on image and data processing of OCT B-scans. By analysing the surface structure, defects, like cuts or tears, are detectable. A further indicator for the quality of the ASE is the morphology of the tissue. This allows to determine if the skin model has reached the final growth state. We found, that OCT is a well suited technology for automatically characterizing artificial skin equivalents and validating the application as testing system.

Using Narratives to Explore Other-Directed Occupational Choice and Academic Success

Science.gov (United States)

Rehfuss, Mark C.; Borges, Nicole J.

2006-01-01

The construct of "other-directed versus self-directed career choice" has existed for quite some time. The current focus of vocational psychologists and counselors has made them question the relevance of this construct for contemporary American society. Many counselors today challenge the assumption that a career choice based on others'…

Continuous "in vitro" Evolution of a Ribozyme Ligase: A Model Experiment for the Evolution of a Biomolecule

Science.gov (United States)

Ledbetter, Michael P.; Hwang, Tony W.; Stovall, Gwendolyn M.; Ellington, Andrew D.

2013-01-01

Evolution is a defining criterion of life and is central to understanding biological systems. However, the timescale of evolutionary shifts in phenotype limits most classroom evolution experiments to simple probability simulations. "In vitro" directed evolution (IVDE) frequently serves as a model system for the study of Darwinian…

Meso and micro level workforce challenges in psychiatric rehabilitation.

Science.gov (United States)

Reifels, Lennart; Pirkis, Jane

2012-01-01

Results of an exploratory study are presented which examined workforce challenges in Australia's most established psychiatric rehabilitation sector. The study had the two-fold aim of investigating workforce challenges at an organizational ("meso") level and at the level of direct-service workers' daily practice ("micro"). Data from 23 key informant interviews conducted with service managers and long-serving staff were analyzed through basic descriptive and thematic analyses. Organizations faced significant annual staff turnover (25.6%), specific staff supply shortages, and challenges in recruiting staff with adequate experience and longevity to match the complexity of client issues. Workers equally encountered challenges in this increasingly complex and rapidly changing field of work. CONCLUSIONS & IMPLICATIONS FOR PRACTICE: Workforce strategies designed to attract/retain experienced staff can improve workforce cohesiveness and sustainability, as can training and support activities aimed at equipping staff to reflect on and operate in dynamic and changing work environments.

High Uric Acid Induces Insulin Resistance in Cardiomyocytes In Vitro and In Vivo.

Directory of Open Access Journals (Sweden)

Li Zhi

Full Text Available Clinical studies have shown hyperuricemia strongly associated with insulin resistance as well as cardiovascular disease. Direct evidence of how high uric acid (HUA affects insulin resistance in cardiomyocytes, but the pathological mechanism of HUA associated with cardiovascular disease remains to be clarified. We aimed to examine the effect of HUA on insulin sensitivity in cardiomyocytes and on insulin resistance in hyperuricemic mouse model. We exposed primary cardiomyocytes and a rat cardiomyocyte cell line, H9c2 cardiomyocytes, to HUA, then quantified glucose uptake with a fluorescent glucose analog, 2-NBDG, after insulin challenge and detected reactive oxygen species (ROS production. Western blot analysis was used to examine the levels of insulin receptor (IR, phosphorylated insulin receptor substrate 1 (IRS1, Ser307 and phospho-Akt (Ser473. We monitored the impact of HUA on insulin resistance, insulin signaling and IR, phospho-IRS1 (Ser307 and phospho-Akt levels in myocardial tissue of an acute hyperuricemia mouse model established by potassium oxonate treatment. HUA inhibited insulin-induced glucose uptake in H9c2 and primary cardiomyocytes. It increased ROS production; pretreatment with N-acetyl-L-cysteine (NAC, a ROS scavenger, reversed HUA-inhibited glucose uptake induced by insulin. HUA exposure directly increased the phospho-IRS1 (Ser307 response to insulin and inhibited that of phospho-Akt in H9C2 cardiomyocytes, which was blocked by NAC. Furthermore, the acute hyperuricemic mice model showed impaired glucose tolerance and insulin tolerance accompanied by increased phospho-IRS1 (Ser307 and inhibited phospho-Akt response to insulin in myocardial tissues. HUA inhibited insulin signaling and induced insulin resistance in cardiomyocytes in vitro and in vivo, which is a novel potential mechanism of hyperuricemic-related cardiovascular disease.

Ultra-accurate collaborative information filtering via directed user similarity

Science.gov (United States)

Guo, Q.; Song, W.-J.; Liu, J.-G.

2014-07-01

A key challenge of the collaborative filtering (CF) information filtering is how to obtain the reliable and accurate results with the help of peers' recommendation. Since the similarities from small-degree users to large-degree users would be larger than the ones in opposite direction, the large-degree users' selections are recommended extensively by the traditional second-order CF algorithms. By considering the users' similarity direction and the second-order correlations to depress the influence of mainstream preferences, we present the directed second-order CF (HDCF) algorithm specifically to address the challenge of accuracy and diversity of the CF algorithm. The numerical results for two benchmark data sets, MovieLens and Netflix, show that the accuracy of the new algorithm outperforms the state-of-the-art CF algorithms. Comparing with the CF algorithm based on random walks proposed by Liu et al. (Int. J. Mod. Phys. C, 20 (2009) 285) the average ranking score could reach 0.0767 and 0.0402, which is enhanced by 27.3% and 19.1% for MovieLens and Netflix, respectively. In addition, the diversity, precision and recall are also enhanced greatly. Without relying on any context-specific information, tuning the similarity direction of CF algorithms could obtain accurate and diverse recommendations. This work suggests that the user similarity direction is an important factor to improve the personalized recommendation performance.

The secretory products of Trichomonas vaginalis decrease fertilizing capacity of mice sperm in vitro

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Jaesook Roh

2015-04-01

Full Text Available Trichomonas vaginalis infection is one of the most prevalent sexually transmitted infections in humans and is now recognized as an important cause of infertility in men. There is little information about the effect of extracellular polymeric substances (EPS from T. vaginalis on sperm, but previous reports do not provide a conclusive description of the functional integrity of the sperm. To investigate the impact of EPS on the fertilizing capacity of sperm, we assessed sperm motility, acrosomal status, hypo-osmotic swelling, and in vitrofertilization rate after incubating the sperm with EPS in vitrousing mice. The incubation of sperm with EPS significantly decreased sperm motility, viability, and functional integrity in a concentration and time-dependent manner. These effects on sperm quality also resulted in a decreased fertilization rate in vitro. This is the first report that demonstrates the direct negative impact of the EPS of T. vaginalis on the fertilization rate of sperm in vitro. However, further study should be performed using human sperm to determine if EPS has similar negative impact on human sperm fertilizing capacity in vitro.

Expressive communication of children with autism: the use of challenging behaviour.

Science.gov (United States)

Chiang, Hsu-Min

2008-11-01

There is a lack of empirical research investigating challenging behaviour in children with autism with severe speech impairments in naturalistic settings. The aim of the present study was to investigate challenging behaviour among Australian and Taiwanese children with autism who are non-verbal or have limited speech (i.e. less than five functional words) in school settings and to address the following questions: (1) What are the communicative characteristics (e.g. communicative functions, partners, activities and partner's responses) of challenging behaviour? (2) Do sample characteristics (e.g. age, severity of autism, adaptive behaviour level, the use of graphic symbols, the use of non-symbolic acts, the use of speech) associate with the amount of challenging behaviour? (3) Do cultural differences exist in communicative variables of challenging behaviour? Naturalistic observations were used to investigate challenging behaviour in children with autism. A total of 32 (17 Australian and 15 Taiwanese) children with autism who were non-verbal or had limited speech participated in the current study and each participant was videotaped for 2 h during his/her everyday school routines. Sixteen out of 32 children had challenging behaviour. Challenging behaviour was used for requesting and rejecting communicative functions. A paired-samples t-test revealed that no significant differences existed in the frequency of occurrence between request and reject. Challenging behaviour was directed to adults and peers. A paired-samples t-test revealed that challenging behaviour was directed to adults significantly more frequently than to peers. Communicative partners showed different responses (e.g. deny, no response, delivered, removed, acknowledge) to challenging behaviour. A repeated-measures analysis of variance (ANOVA) revealed a main effect for partner response. Deny was the most common response. Challenging behaviour occurred across a variety of classroom activities (e

Effect of in vitro irradiation and cell cycle-inhibitory drugs on the spontaneous human IgE synthesis in vitro

International Nuclear Information System (INIS)

Del Prete, G.F.; Vercelli, D.; Tiri, A.; Maggi, E.; Rossi, O.; Romagnani, S.; Ricci, M.

1987-01-01

The in vitro effects of radiation, diterpine forskolin (FK), and hydrocortisone (HC) on the in vitro spontaneous IgE synthesis by peripheral blood B-lymphocytes from atopic patients were investigated. Without affecting cell viability, in vitro irradiation inhibited in a dose-dependent fashion de novo IgE synthesis in vitro by B cells from all patients examined with a mean 40% reduction of in vitro IgE product after treatment with 100 rads. In contrast, the in vitro IgE production by the U266 myeloma cell line was unaffected, even by irradiation with 1600 rads. The addition to B cell cultures from atopic patients of FK consistently resulted in a dose-dependent inhibition of the spontaneous IgE production in vitro. The addition to cultures of 10(-5) and 10(-6) molar concentrations of HC was also usually inhibitory, whereas lower HC concentrations were uneffective or even enhanced the spontaneous in vitro IgE synthesis. When 10(-6) molar concentrations of both HC and FK were combined in culture, a summation inhibitory effect on the spontaneous IgE synthesis was observed. In contrast, neither FK nor HC had inhibitory effect on the in vitro spontaneous IgE synthesis by the U266 myeloma cell line. The spontaneous in vitro IgE synthesis by B cells from patients with Hodgkin's disease, demonstrating high levels of serum IgE, was strongly reduced or virtually abolished after patients underwent total nodal irradiation to prevent the spread of the disease. In addition, the in vitro spontaneous IgE synthesis by B cells from atopic patients was markedly decreased or abolished by in vivo administration of betamethasone

In vitro cytotoxic screening of selected Saudi medicinal plants.

Science.gov (United States)

Almehdar, Hussein; Abdallah, Hossam M; Osman, Abdel-Moneim M; Abdel-Sattar, Essam A

2012-04-01

Many natural products from plants have been identified to exert anticancer activity. It might be expected to be a challenge to look at the Saudi plants in order to discover new sources for new molecules which may have anticancer activity. The methanolic extracts of forty species of plants traditionally used in Saudi Arabia for the treatment of a variety of diseases were tested in vitro for their potential anticancer activity on different human cancer cell lines. The cytotoxic activity of the methanolic extracts of the tested plants were determined using three human cancer cell lines, namely, breast cancer (MCF7), hepatocellular carcinoma (HEPG2), and cervix cancer (HELA) cells. In addition, human normal melanocyte (HFB4) was used as normal nonmalignant cells. Sulforhodamine B colorimetric assay was used to evaluate the in vitro cytotoxic activity of the different extracts. The growth inhibition of 50% (IC(50)) for each extract was calculated from the optical density of treated and untreated cells. Doxorubicin, a broad-spectrum anticancer drug, was used as the positive control. Nine plant extracts were chosen for further fractionation based on their activity and availability. Interesting cytotoxic activity was observed for Hypoestes forskaolii, Withania somnifera, Solanum glabratum, Adenium obesum, Pistacia vera oleoresin, Caralluma quadrangula, Eulophia petersii, Phragmanthera austroarabica, and Asparagus officinalis. Other extracts showed poor activity.

Practical challenges related to point of care testing

Directory of Open Access Journals (Sweden)

Julie L.V. Shaw

2016-04-01

Full Text Available Point of care testing (POCT refers to laboratory testing that occurs near to the patient, often at the patient bedside. POCT can be advantageous in situations requiring rapid turnaround time of test results for clinical decision making. There are many challenges associated with POCT, mainly related to quality assurance. POCT is performed by clinical staff rather than laboratory trained individuals which can lead to errors resulting from a lack of understanding of the importance of quality control and quality assurance practices. POCT is usually more expensive than testing performed in the central laboratory and requires a significant amount of support from the laboratory to ensure the quality testing and meet accreditation requirements.Here, specific challenges related to POCT compliance with accreditation standards are discussed along with strategies that can be used to overcome these challenges. These areas include: documentation of POCT orders, charting of POCT results as well as training and certification of individuals performing POCT. Factors to consider when implementing connectivity between POCT instruments and the electronic medical record are also discussed in detail and include: uni-directional versus bidirectional communication, linking patient demographic information with POCT software, the importance of positive patient identification and considering where to chart POCT results in the electronic medical record. Keywords: Point of care Testing, Laboratory accreditation, Medical directive, Results documentation, Electronic Medical Record, Transcription error, Connectivity, Positive patient identification

Safety Evaluation of Cosmetic Ingredients: In Vitro Opportunities for the Identification of Contact Allergens

Directory of Open Access Journals (Sweden)

Emanuela Corsini

2014-03-01

Full Text Available Irritant and allergic contact dermatitis are undesired side effects in the development of drugs and cosmetics as well as after contact with environmental or industrial chemicals. Over the last decades, a great deal of progress has been made in the development of alternative In vitro test to assess these issues. Driven by the 7th Amendment to the European Cosmetic Directive, the EU policy on chemicals (the registration, evaluation, authorization and restriction of chemicals (REACH system, the update of the European legislation on the protection of animals used in research, and emerging visions and strategies for predicting toxicity, in vitro methods are likely to play a major role in the near future. On 12 December 2013, the European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM, part of the European Commission Joint Research Centre published its Recommendation on the Direct Peptide Reactivity Assay (DPRA for skin sensitization, capable of distinguishing sensitizers from non-sensitizers. Other assays (i.e., KeratinoSens™ assay will follow shortly. While a number of methods are at various stages of development and use, currently it is not possible to rank chemicals for their sensitizing potency, an issue that is important for a full safety assessment. It is expected that a predictive method to totally replace animal testing will be in the form of a test battery comprising molecular, cell-based, and/or computational methods, the so-called “Integrated Approaches to Testing and Assessment”. This review aims to discuss the state-of-the-art in the field of in vitro assessment of contact sensitizers.

An efficient in vitro shoot regeneration from leaf petiolar explants and ex vitro rooting of Bixa orellana L.- A dye yielding plant.

Science.gov (United States)

Mohammed, Arifullah; Chiruvella, Kishore K; Namsa, Nima D; Ghanta, Rama Gopal

2015-07-01

Bixa orellana L. (Bixaceae) is a multipurpose tree grown for the production of commercially important dyes. In the present study, an efficient, reproducible protocol was developed for direct plant regeneration from in vitro derived petiole explants of Bixa orellana L. Murashige and Skoog medium (MS) supplemented with 2-isopentenyl adenine (9.8 μM) and naphthalene acetic acid (10.7 μM) was found to be optimum for production of high frequency of shoot organogenesis. Subculturing of the shoots onto the fresh MS medium containing similar concentrations of 2-iP (9.8 μM) and NAA (10.7 μM) produced elongated shoots. Elongated shoots when placed onto MS medium supplemented with 1.7 μM indole-3-acetic acid and 14.7 μM 2-iP produced optimal rooting. Rooted plantlets were acclimatized and transplanted to the field successfully. Histological investigation revealed the origin of shoot primordia, from sub-epidermal cells of petiole explants. The regeneration protocol developed in this study can be useful for mass in vitro propagation and effective genetic transformation of commercially important edible dye yielding tree species.

Exploring the Challenges of Evidence-based HIV and AIDS ...

African Journals Online (AJOL)

The study surveyed the challenges associated with HIV prevention technologies highlighting both positive indicators of success and negative indicators of technology failure. Attendant health risks associated with prevention failure are discussed. Direction for future studies could focus on HIV risk reduction in clinical trials.

[Challenges of Digital Medicine].

Science.gov (United States)

Blaser, Jürg

2018-06-01

Challenges of Digital Medicine Abstract. Digitization is increasingly covering more and more sectors, including medicine. To ensure medical operation 365 × 24 hours, progressively more human and financial resources are necessary. The transformation of patient histories from paper into electronic patient records focused initially on documentation. Today, hospital information systems are increasingly used as a platform for the communication of all professionals involved in the patient process - in Switzerland, however, so far without providing patients direct access to their data. Digititizing processes intend to increase efficiency, but also to enhance clinical and administrative decision support and quality assurance. The introduction of the electronic patient record in Switzerland in 2020 is expected to provide cross-company, more complete documentation of patient care. Multimorbid patients, often treated in different institutions and by different specialists, should benefit from this in particular. Advances in artificial intelligence offer new opportunities in medicine. Challenges include ensuring reliable data protection, and better interoperability of the systems involved. Semantically structured, machine-readable data exchange is a necessity for both networked services and internationally competitive research.

Developing a Strong Direct Care Workforce.

Science.gov (United States)

Stear, Lynda A

The aging population of the United States has led to a need for more direct care workers to provide personal care to older adults and disabled people in their homes. The U.S. Bureau of Labor Statistics predicts employment of paraprofessionals in home care will grow 49% between 2012 and 2022 as the baby boomer generation ages and requires more assistance in activities of daily living. The typical direct care worker is a female aged 25 to 54 years old, a demographic that is projected to remain flat in the coming years. Direct care workers typically are poorly paid for work that is labor-intensive and often work in less than optimal working conditions. It is important that agencies hiring direct care workers understand the challenges these workers face and institute sound hiring practices and provide proper training and ongoing supervision. It is possible to have a quality direct care team. Proper training, role modeling, and supervision will improve employee satisfaction, decrease turnover, and improve care outcomes for patients.

The implementation of the EU renewable directive in Spain. Strategies and challenges

International Nuclear Information System (INIS)

Labriet, Maryse; Cabal, Helena; Lechon, Yolanda; Giannakidis, George; Kanudia, Amit

2010-01-01

Based on the European project RES2020, the analysis evaluates the energy strategies to be implemented in Spain in order to satisfy the EU Renewable Directive. The modelling framework relies on the technico-economic model TIMES-Spain, part of the Pan-European TIMES model used in the project. TIMES is a bottom-up technology rich optimisation model representing the whole energy systems of the countries. Among the results, it appears that the gap regarding the renewable deployment in Spain between the Business-as-Usual case (including the existing policies) and the EU Directive should be compensated mainly by the penetration of bioenergy in transport and industry, and by the implementation of conservation measures, which contribute to reduce the total energy demand and thus makes useless additional investments in renewable power plants compared to the Business-as-Usual case. Only higher climate mitigation ambitions result in an absolute increase in the renewable-based electricity generation compared to the Business-as-Usual case. Moreover, when allowed, Spain is offering renewable energy credits under the statistical transfer mechanism to other European countries. The cost increase of the modelled renewable and climate policies compared to the Business-as-Usual remains relatively minor.

Bidirectional interactions between neuronal and hemodynamic responses to transcranial direct current stimulation (tDCS: challenges for brain-state dependent tDCS

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Anirban eDutta

2015-08-01

Full Text Available Transcranial direct current stimulation (tDCS has been shown to modulate cortical neural activity. During neural activity, the electric currents from excitable membranes of brain tissue superimpose in the extracellular medium and generate a potential at scalp, which is referred as the electroencephalogram (EEG. Respective neural activity (energy demand has been shown to be closely related, spatially and temporally, to cerebral blood flow (CBF that supplies glucose (energy supply via neurovascular coupling. The hemodynamic response can be captured by near-infrared spectroscopy (NIRS, which enables continuous monitoring of cerebral oxygenation and blood volume. This neurovascular coupling phenomenon led to the concept of neurovascular unit (NVU that consists of the endothelium, glia, neurons, pericytes, and the basal lamina. Here, recent works suggest NVU as an integrated system working in concert using feedback mechanisms to enable proper brain homeostasis and function where the challenge remains in capturing these mostly nonlinear spatiotemporal interactions within NVU during tDCS. Therefore, we propose EEG-NIRS-based whole-head monitoring of tDCS-induced neuronal and hemodynamic alterations for brain-state dependent tDCS.

Directed and persistent movement arises from mechanochemistry of the ParA/ParB system.

Science.gov (United States)

Hu, Longhua; Vecchiarelli, Anthony G; Mizuuchi, Kiyoshi; Neuman, Keir C; Liu, Jian

2015-12-22

The segregation of DNA before cell division is essential for faithful genetic inheritance. In many bacteria, segregation of low-copy number plasmids involves an active partition system composed of a nonspecific DNA-binding ATPase, ParA, and its stimulator protein ParB. The ParA/ParB system drives directed and persistent movement of DNA cargo both in vivo and in vitro. Filament-based models akin to actin/microtubule-driven motility were proposed for plasmid segregation mediated by ParA. Recent experiments challenge this view and suggest that ParA/ParB system motility is driven by a diffusion ratchet mechanism in which ParB-coated plasmid both creates and follows a ParA gradient on the nucleoid surface. However, the detailed mechanism of ParA/ParB-mediated directed and persistent movement remains unknown. Here, we develop a theoretical model describing ParA/ParB-mediated motility. We show that the ParA/ParB system can work as a Brownian ratchet, which effectively couples the ATPase-dependent cycling of ParA-nucleoid affinity to the motion of the ParB-bound cargo. Paradoxically, this resulting processive motion relies on quenching diffusive plasmid motion through a large number of transient ParA/ParB-mediated tethers to the nucleoid surface. Our work thus sheds light on an emergent phenomenon in which nonmotor proteins work collectively via mechanochemical coupling to propel cargos-an ingenious solution shaped by evolution to cope with the lack of processive motor proteins in bacteria.

Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER.

Science.gov (United States)

Dodds, E; Dunckley, M G; Naujoks, K; Michaelis, U; Dickson, G

1998-04-01

Cationic lipid-DNA complexes (lipoplexes) have been widely used as gene transfer vectors which avoid the adverse immunogenicity and potential for viraemia of viral vectors. With the long-term aim of gene transfer into skeletal muscle in vivo, we describe a direct in vitro comparison of two commercially available cationic lipid formulations, Lipofectamine and DOSPER. Optimisation of transfection was performed in the C2C12 mouse muscle cell line, before further studies in primary mouse myoblasts and C2C12 myotubes. Reporter gene constructs expressing either E. coli beta-galactosidase or green fluorescent protein (GFP) were used in order to evaluate transfection efficiency by histochemical staining or FACS analysis, respectively. Both lipid formulations were able to promote efficient, reproducible gene transfer in C2C12 cells, and to transfect primary mouse myoblast cultures successfully. However, DOSPER exhibited the important advantage of being able to transfect cells in the presence of serum of both bovine and murine origin. This feature allowed increased cell survival during in vitro transfections, and may be advantageous for direct in vivo gene transfer efficacy.

Estimating challenge load due to disease outbreaks and other challenges using reproduction records of sows.

Science.gov (United States)

Mathur, P K; Herrero-Medrano, J M; Alexandri, P; Knol, E F; ten Napel, J; Rashidi, H; Mulder, H A

2014-12-01

A method was developed and tested to estimate challenge load due to disease outbreaks and other challenges in sows using reproduction records. The method was based on reproduction records from a farm with known disease outbreaks. It was assumed that the reduction in weekly reproductive output within a farm is proportional to the magnitude of the challenge. As the challenge increases beyond certain threshold, it is manifested as an outbreak. The reproduction records were divided into 3 datasets. The first dataset called the Training dataset consisted of 57,135 reproduction records from 10,901 sows from 1 farm in Canada with several outbreaks of porcine reproductive and respiratory syndrome (PRRS). The known disease status of sows was regressed on the traits number born alive, number of losses as a combination of still birth and mummified piglets, and number of weaned piglets. The regression coefficients from this analysis were then used as weighting factors for derivation of an index measure called challenge load indicator. These weighting factors were derived with i) a two-step approach using residuals or year-week solutions estimated from a previous step, and ii) a single-step approach using the trait values directly. Two types of models were used for each approach: a logistic regression model and a general additive model. The estimates of challenge load indicator were then compared based on their ability to detect PRRS outbreaks in a Test dataset consisting of records from 65,826 sows from 15 farms in the Netherlands. These farms differed from the Canadian farm with respect to PRRS virus strains, severity and frequency of outbreaks. The single-step approach using a general additive model was best and detected 14 out of the 15 outbreaks. This approach was then further validated using the third dataset consisting of reproduction records of 831,855 sows in 431 farms located in different countries in Europe and America. A total of 41 out of 48 outbreaks detected

The post-2015 development agenda for diabetes in sub-Saharan Africa: challenges and future directions.

Science.gov (United States)

Renzaho, Andre M N

2015-01-01

Diabetes is one of the non-communicable diseases (NCDs) which is rising significantly across sub-Saharan African (SSA) countries and posing a threat to the social, economic, and cultural fabric of the SSA population. The inclusion of NCDs into the post-2015 development agenda along with the global monitoring framework provides an opportunity to monitor progress of development programmes in developing countries. This paper examines challenges associated with dealing with diabetes within the development agenda in SSA and explores some policy options. This conceptual review draws from a range of works published in Medline and the grey literature to advance the understanding of the post-2015 development agenda and how it relates to NCDs. The paper begins with the burden of diabetes in sub-Sahara Africa and then moves on to examine challenges associated with diabetes prevention, treatment, and management in Africa. It finishes by exploring policy implications. With regards to development programmes on NCDs in the SSA sub-continent, several challenges exist: 1) poor documentation of risk factors, 2) demographic transitions (rapid urbanisation and ageing), 3) the complementary role of traditional healers, 4) tuberculosis and the treatment of the acquired immunodeficiency syndrome as risk factors for diabetes, 5) diabetes in complex emergencies, 6) diabetes as an international development priority and not a policy agenda for many SSA countries, and 7) poorly regulated food and beverage industry. For the post-2015 development agenda for NCDs to have an impact, sufficient investments will be needed to address legislative, technical, human, and fiscal resource constraints through advocacy, accountability, political leadership, and effective public-private partnership. Striking the right balance between competing demands and priorities, policies, and implementation strategies hold the key to an effective response to diabetes in SSA countries.

The post-2015 development agenda for diabetes in sub-Saharan Africa: challenges and future directions

Directory of Open Access Journals (Sweden)

Andre M. N. Renzaho

2015-05-01

Full Text Available Background: Diabetes is one of the non-communicable diseases (NCDs which is rising significantly across sub-Saharan African (SSA countries and posing a threat to the social, economic, and cultural fabric of the SSA population. The inclusion of NCDs into the post-2015 development agenda along with the global monitoring framework provides an opportunity to monitor progress of development programmes in developing countries. This paper examines challenges associated with dealing with diabetes within the development agenda in SSA and explores some policy options. Design: This conceptual review draws from a range of works published in Medline and the grey literature to advance the understanding of the post-2015 development agenda and how it relates to NCDs. The paper begins with the burden of diabetes in sub-Sahara Africa and then moves on to examine challenges associated with diabetes prevention, treatment, and management in Africa. It finishes by exploring policy implications. Results: With regards to development programmes on NCDs in the SSA sub-continent, several challenges exist: 1 poor documentation of risk factors, 2 demographic transitions (rapid urbanisation and ageing, 3 the complementary role of traditional healers, 4 tuberculosis and the treatment of the acquired immunodeficiency syndrome as risk factors for diabetes, 5 diabetes in complex emergencies, 6 diabetes as an international development priority and not a policy agenda for many SSA countries, and 7 poorly regulated food and beverage industry. Conclusion: For the post-2015 development agenda for NCDs to have an impact, sufficient investments will be needed to address legislative, technical, human, and fiscal resource constraints through advocacy, accountability, political leadership, and effective public–private partnership. Striking the right balance between competing demands and priorities, policies, and implementation strategies hold the key to an effective response to diabetes

Towards the Identification of an In Vitro Tool for Assessing the Biological Behavior of Aerosol Supplied Nanomaterials

Directory of Open Access Journals (Sweden)

Luisana Di Cristo

2018-03-01

Full Text Available Nanoparticles (NP-based inhalation systems for drug delivery can be administered in liquid form, by nebulization or using pressurized metered dose inhalers, and in solid form by means of dry powder inhalers. However, NP delivery to the lungs has many challenges including the formulation instability due to particle-particle interactions and subsequent aggregation, causing poor deposition in the small distal airways and subsequent alveolar macrophages activity, which could lead to inflammation. This work aims at providing an in vitro experimental design for investigating the correlation between the physico-chemical properties of NP, and their biological behavior, when they are used as NP-based inhalation treatments, comparing two different exposure systems. By means of an aerosol drug delivery nebulizer, human lung cells cultured at air–liquid interface (ALI were exposed to two titanium dioxide NP (NM-100 and NM-101, obtained from the JRC repository. In parallel, ALI cultures were exposed to NP suspension by direct inoculation, i.e., by adding the NP suspensions on the apical side of the cell cultures with a pipette. The formulation stability of NP, measured as hydrodynamic size distributions, the cell viability, cell monolayer integrity, cell morphology and pro-inflammatory cytokines secretion were investigated. Our results demonstrated that the formulation stability of NM-100 and NM-101 was strongly dependent on the aggregation phenomena that occur in the conditions adopted for the biological experiments. Interestingly, comparable biological data between the two exposure methods used were observed, suggesting that the conventional exposure coupled to ALI culturing conditions offers a relevant in vitro tool for assessing the correlation between the physico-chemical properties of NP and their biological behavior, when NP are used as drug delivery systems.

Evaluation of peroxidative stress of cancer cells in vitro by real-time quantification of volatile aldehydes in culture headspace.

Science.gov (United States)

Shestivska, Violetta; Rutter, Abigail V; Sulé-Suso, Josep; Smith, David; Španěl, Patrik

2017-08-30

Peroxidation of lipids in cellular membranes results in the release of volatile organic compounds (VOCs), including saturated aldehydes. The real-time quantification of trace VOCs produced by cancer cells during peroxidative stress presents a new challenge to non-invasive clinical diagnostics, which as described here, we have met with some success. A combination of selected ion flow tube mass spectrometry (SIFT-MS), a technique that allows rapid, reliable quantification of VOCs in humid air and liquid headspace, and electrochemistry to generate reactive oxygen species (ROS) in vitro has been used. Thus, VOCs present in the headspace of CALU-1 cancer cell line cultures exposed to ROS have been monitored and quantified in real time using SIFT-MS. The CALU-1 lung cancer cells were cultured in 3D collagen to mimic in vivo tissue. Real-time SIFT-MS analyses focused on the volatile aldehydes: propanal, butanal, pentanal, hexanal, heptanal and malondialdehyde (propanedial), that are expected to be products of cellular membrane peroxidation. All six aldehydes were identified in the culture headspace, each reaching peak concentrations during the time of exposure to ROS and eventually reducing as the reactants were depleted in the culture. Pentanal and hexanal were the most abundant, reaching concentrations of a few hundred parts-per-billion by volume, ppbv, in the culture headspace. The results of these experiments demonstrate that peroxidation of cancer cells in vitro can be monitored and evaluated by direct real-time analysis of the volatile aldehydes produced. The combination of adopted methodology potentially has value for the study of other types of VOCs that may be produced by cellular damage. Copyright © 2017 John Wiley & Sons, Ltd.

Directed evolution of an LBP/CD14 inhibitory peptide and its anti-endotoxin activity.

Directory of Open Access Journals (Sweden)

Li Fang

Full Text Available BACKGROUND: LPS-binding protein (LBP and its ligand CD14 are located upstream of the signaling pathway for LPS-induced inflammation. Blocking LBP and CD14 binding might prevent LPS-induced inflammation. In previous studies, we obtained a peptide analog (MP12 for the LBP/CD14 binding site and showed that this peptide analog had anti-endotoxin activity. In this study, we used in vitro directed evolution for this peptide analog to improve its in vivo and in vitro anti-endotoxin activity. METHODS: We used error-prone PCR (ep-PCR and induced mutations in the C-terminus of LBP and attached the PCR products to T7 phages to establish a mutant phage display library. The positive clones that competed with LBP for CD14 binding was obtained by screening. We used both in vivo and in vitro experiments to compare the anti-endotoxin activities of a polypeptide designated P1 contained in a positive clone and MP12. RESULTS: 11 positive clones were obtained from among target phages. Sequencing showed that 9 positive clones had a threonine (T to methionine (M mutation in amino acid 287 of LBP. Compared to polypeptide MP12, polypeptide P1 significantly inhibited LPS-induced TNF-α expression and NF-κB activity in U937 cells (P<0.05. Compared to MP12, P1 significantly improved arterial oxygen pressure, an oxygenation index, and lung pathology scores in LPS-induced ARDS rats (P<0.05. CONCLUSION: By in vitro directed evolution of peptide analogs for the LBP/CD14 binding site, we established a new polypeptide (P1 with a threonine (T-to-methionine (M mutation in amino acid 287 of LBP. This polypeptide had high anti-endotoxin activity in vitro and in vivo, which suggested that amino acid 287 in the C-terminus of LBP may play an important role in LBP binding with CD14.

In vitro effects of direct current electric fields on adipose-derived stromal cells.

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Hammerick, Kyle E; Longaker, Michael T; Prinz, Fritz B

2010-06-18

Endogenous electric fields play an important role in embryogenesis, regeneration, and wound repair and previous studies have shown that many populations of cells, leukocytes, fibroblasts, epithelial cells, and endothelial cells, exhibit directed migration in response to electric fields. As regenerative therapies continue to explore ways to control mesenchymal progenitor cells to recreate desirable tissues, it is increasingly necessary to characterize the vast nature of biological responses imposed by physical phenomena. Murine adipose-derived stromal cells (mASCs) migrated toward the cathode in direct current (DC) fields of physiologic strength and show a dose dependence of migration rate to stronger fields. Electric fields also caused mASCs to orient perpendicularly to the field vector and elicited a transient increase in cytosolic calcium. Additionally, their galvanotactic response appears to share classic chemotactic signaling pathways that are involved in the migration of other cell types. Galvanotaxis is one predominant result of electric fields on mASCs and it may be exploited to engineer adult stem cell concentrations and locations within implanted grafts or toward sites of wound repair. Copyright (c) 2010 Elsevier Inc. All rights reserved.

An extensive cocktail approach for rapid risk assessment of in vitro CYP450 direct reversible inhibition by xenobiotic exposure

International Nuclear Information System (INIS)

Spaggiari, Dany; Daali, Youssef; Rudaz, Serge

2016-01-01

Acute exposure to environmental factors strongly affects the metabolic activity of cytochrome P450 (P450). As a consequence, the risk of interaction could be increased, modifying the clinical outcomes of a medication. Because toxic agents cannot be administered to humans for ethical reasons, in vitro approaches are therefore essential to evaluate their impact on P450 activities. In this work, an extensive cocktail mixture was developed and validated for in vitro P450 inhibition studies using human liver microsomes (HLM). The cocktail comprised eleven P450-specific probe substrates to simultaneously assess the activities of the following isoforms: 1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 2J2 and subfamily 3A. The high selectivity and sensitivity of the developed UHPLC-MS/MS method were critical for the success of this methodology, whose main advantages are: (i) the use of eleven probe substrates with minimized interactions, (ii) a low HLM concentration, (iii) fast incubation (5 min) and (iv) the use of metabolic ratios as microsomal P450 activities markers. This cocktail approach was successfully validated by comparing the obtained IC 50 values for model inhibitors with those generated with the conventional single probe methods. Accordingly, reliable inhibition values could be generated 10-fold faster using a 10-fold smaller amount of HLM compared to individual assays. This approach was applied to assess the P450 inhibition potential of widespread insecticides, namely, chlorpyrifos, fenitrothion, methylparathion and profenofos. In all cases, P450 2B6 was the most affected with IC 50 values in the nanomolar range. For the first time, mixtures of these four insecticides incubated at low concentrations showed a cumulative inhibitory in vitro effect on P450 2B6. - Highlights: • Ten P450 isoforms activities assessed simultaneously with only one incubation. • P450 activity levels measured using the metabolic ratio approach. • IC 50 values generated 10-fold faster

A Novel Biodegradable Polyurethane Matrix for Auricular Cartilage Repair: An In Vitro and In Vivo Study.

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Iyer, Kartik; Dearman, Bronwyn L; Wagstaff, Marcus J D; Greenwood, John E

2016-01-01

Auricular reconstruction poses a challenge for reconstructive and burns surgeons. Techniques involving cartilage tissue engineering have shown potential in recent years. A biodegradable polyurethane matrix developed for dermal reconstruction offers an alternative to autologous, allogeneic, or xenogeneic biologicals for cartilage reconstruction. This study assesses such a polyurethane matrix for this indication in vivo and in vitro. To evaluate intrinsic cartilage repair, three pigs underwent auricular surgery to create excisional cartilage ± perichondrial defects, measuring 2 × 3 cm in each ear, into which acellular polyurethane matrices were implanted. Biopsies were taken at day 28 for histological assessment. Porcine chondrocytes ± perichondrocytes were cultured and seeded in vitro onto 1 × 1 cm polyurethane scaffolds. The total culture period was 42 days; confocal, histological, and immunohistochemical analyses of scaffold cultures were performed on days 14, 28, and 42. In vivo, the polyurethane matrices integrated with granulation tissue filling all biopsy samples. Minimal neocartilage invasion was observed marginally on some samples. Tissue composition was identical between ears whether perichondrium was left intact, or not. In vitro, the polyurethane matrix was biocompatible with chondrocytes ± perichondrocytes and supported production of extracellular matrix and Type II collagen. No difference was observed between chondrocyte culture alone and chondrocyte/perichondrocyte scaffold coculture. The polyurethane matrix successfully integrated into the auricular defect and was a suitable scaffold in vitro for cartilage tissue engineering, demonstrating its potential application in auricular reconstruction.

In vitro incorporation of the phage Phi29 connector complex

International Nuclear Information System (INIS)

Fu Chiyu; Prevelige, Peter E.

2009-01-01

The incorporation of the DNA packaging connector complex during lambdoid phage assembly in vivo is strictly controlled-one and only one of the twelve identical icosahedral vertices is differentiated by the inclusion of a portal or connector dodecamer. Proposed control mechanisms include obligate nucleation from a connector containing complex, addition of the connector as the final step during assembly, and a connector-mediated increase in the growth rate. The inability to recapitulate connector incorporation in vitro has made it difficult to obtain direct biochemical evidence in support of one model over another. Here we report the development an in vitro assembly system for the well characterized dsDNA phage Phi29 which results in the co-assembly of connector with capsid and scaffolding proteins to form procapsid-like particles (PLPs). Immuno-electron microscopy demonstrates the specific incorporation of connector vertex in PLPs. The connector protein increases both the yield and the rate of capsid assembly suggesting that the incorporation of the connector in Phi29 likely promotes nucleation of assembly.

Population dynamics and in vitro antibody pressure of porcine parvovirus indicate a decrease in variability.

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Streck, André Felipe; Homeier, Timo; Foerster, Tessa; Truyen, Uwe

2013-09-01

To estimate the impact of porcine parvovirus (PPV) vaccines on the emergence of new phenotypes, the population dynamic history of the virus was calculated using the Bayesian Markov chain Monte Carlo method with a Bayesian skyline coalescent model. Additionally, an in vitro model was performed with consecutive passages of the 'Challenge' strain (a virulent field strain) and NADL2 strain (a vaccine strain) in a PK-15 cell line supplemented with polyclonal antibodies raised against the vaccine strain. A decrease in genetic diversity was observed in the presence of antibodies in vitro or after vaccination (as estimated by the in silico model). We hypothesized that the antibodies induced a selective pressure that may reduce the incidence of neutral selection, which should play a major role in the emergence of new mutations. In this scenario, vaccine failures and non-vaccinated populations (e.g. wild boars) may have an important impact in the emergence of new phenotypes.

Radiosynthesis and in vitro evaluation of 99mTc(CO)3-labeled folic acid derivative

International Nuclear Information System (INIS)

Drishty Satpati; Archana Mukherjee; Meera Venkatesh; Sharmila Banerjee

2011-01-01

The over-expression of folate receptors in variety of neoplastic tissues makes radiolabeled folate conjugates potential agents for imaging and therapy of such cancers. With the aim of preparing an imaging agent for targeting folate receptors, folic acid has been conjugated with homocysteine for complexation with [ 99m Tc(CO) 3 (H 2 O) 3 ] + core. The radiolabeled complex of the homocysteine-folate could be obtained in >95% radiochemical yield as observed by HPLC. Stability of complex in saline was studied and challenge studies with histidine and cysteine revealed kinetic stability of the complex. Lipophilicity of the radiolabeled complex (log P) was found to be 0.45. In vitro uptake of 99m Tc(CO) 3 -labeled folic acid derivative was studied in KB cells and inhibition studies were carried out using 3 H-folic acid and cold homocysteine-folate conjugate. The in vitro studies indicated loss of binding affinity of the derivative towards folate receptors. (author)

Bed Bug Epidemic: A Challenge to Public Health

Science.gov (United States)

Ratnapradipa, Dhitinut; Ritzel, Dale O.; Haramis, Linn D.; Bliss, Kadi R.

2011-01-01

In recent years, reported cases of bed bug infestations in the U.S. and throughout the world have escalated dramatically, posing a global public health problem. Although bed bugs are not known to transmit disease to humans, they pose both direct and indirect public health challenges in terms of health effects, treatment, cost, and resource…

Renewable sources electric power: resources and challenges for the France

International Nuclear Information System (INIS)

Bouchereau, J.M.; Dormoy, C.

2001-05-01

This paper provides information (statistical data, legal framework) on the electric power produced by the renewable energy sources in France. It explains the associated local economical challenge and the french objectives in the European Union Directive. (A.L.B.)

Towards Organs on Demand: Breakthroughs and Challenges in Models of Organogenesis.

Science.gov (United States)

Francipane, Maria Giovanna; Lagasse, Eric

2016-09-01

In recent years, functional three-dimensional (3D) tissue generation in vitro has been significantly advanced by tissue-engineering methods, achieving better reproduction of complex native organs compared to conventional culture systems. This review will discuss traditional 3D cell culture techniques as well as newly developed technology platforms. These recent techniques provide new possibilities in the creation of human body parts and provide more accurate predictions of tissue response to drug and chemical challenges. Given the rapid advancement in the human induced pluripotent stem cell (iPSC) field, these platforms also hold great promise in the development of patient-specific, transplantable tissues and organs on demand.

Direct evaluation of thyroid 127I and iodine overload: in vivo study by X-ray fluorescence and in vitro by SIMS microscopy

International Nuclear Information System (INIS)

Briancon, C.; Jeusset, J.; Halpern, S.; Fragu, P.

1992-01-01

This review describes the two methods which allow direct estimation of stable iodine ( 127 I) within thyroid gland either in vivo by X-ray fluorescence or in vitro by secondary ion mass spectrometry (SIMS) microscopy on tissue section. Although the measurement of thyroid iodine content (TIC) by X-ray fluorescence has little relevance for routine explorations of thyroid function, this is a valuable method for understanding complex pathophysiological conditions such as the thyroid adaptation to iodine overload. On the other hand, SIMS microscopy which is able to characterize the functional activity of thyroid tissue by measuring 127 I concentration within the thyroid follicles, can be used to determine the extent to which exogeneous iodine affects the regulation of iodine within the thyroid follicles. Both methods were used to evaluate the quantitative changes in thyroid 127 I induced by amiodarone iodine overload. TIC measurements shows that hyperthyroidism occured only in patients who increased their iodine stores, while the patients who developed hypothyroidism has low iodine stores. The SIMS microscopy data obtained in mice demonstrated that the thyroid response to amiodarone is related to dietary iodine intake leading to an increase in local iodine concentration in iodine deficient mice and to a decrease in iodine supplemented mice. This response is specific and different from that induced by an iodine overload. These results could explain that hyperthyroidism with high thyroid iodine content occured in areas with low thyroid iodine content in areas with a supplemented iodine diet

In vitro toxicological nanoparticle studies under flow exposure

International Nuclear Information System (INIS)

Sambale, Franziska; Stahl, Frank; Bahnemann, Detlef; Scheper, Thomas

2015-01-01

The use of nanoparticles is becoming increasingly common in industry and everyday objects. Thus, extensive risk management concerning the potential health risk of nanoparticles is important. Currently, in vitro nanoparticle testing is mainly performed under static culture conditions without any shear stress. However, shear stress is an important biomechanical parameter. Therefore, in this study, a defined physiological flow to different mammalian cell lines such as A549 cells and NIH-3T3 cells has been applied. The effects of zinc oxide and titanium dioxide nanoparticles (TiO 2 -NP), respectively, were investigated under both static and dynamic conditions. Cell viability, cell morphology, and adhesion were proven and compared to the static cell culture. Flow exposure had an impact on the cellular morphology of the cells. NIH-3T3 cells were elongated in the direction of flow and A549 cells exhibited vesicles inside the cells. Zinc oxide nanoparticles reduced the cell viability in the static and in the dynamic culture; however, the dynamic cultures were more sensitive. In the static culture and in the dynamic culture, TiO 2 -NP did not affect cell viability. In conclusion, dynamic culture conditions are important for further in vitro investigations and provide more relevant results than static culture conditions

In vitro toxicological nanoparticle studies under flow exposure

Energy Technology Data Exchange (ETDEWEB)

Sambale, Franziska, E-mail: sambale@iftc.uni-hannover.de; Stahl, Frank; Bahnemann, Detlef; Scheper, Thomas [Gottfried Wilhelm Leibniz University Hanover, Institute for Technical Chemistry (Germany)

2015-07-15

The use of nanoparticles is becoming increasingly common in industry and everyday objects. Thus, extensive risk management concerning the potential health risk of nanoparticles is important. Currently, in vitro nanoparticle testing is mainly performed under static culture conditions without any shear stress. However, shear stress is an important biomechanical parameter. Therefore, in this study, a defined physiological flow to different mammalian cell lines such as A549 cells and NIH-3T3 cells has been applied. The effects of zinc oxide and titanium dioxide nanoparticles (TiO{sub 2}-NP), respectively, were investigated under both static and dynamic conditions. Cell viability, cell morphology, and adhesion were proven and compared to the static cell culture. Flow exposure had an impact on the cellular morphology of the cells. NIH-3T3 cells were elongated in the direction of flow and A549 cells exhibited vesicles inside the cells. Zinc oxide nanoparticles reduced the cell viability in the static and in the dynamic culture; however, the dynamic cultures were more sensitive. In the static culture and in the dynamic culture, TiO{sub 2}-NP did not affect cell viability. In conclusion, dynamic culture conditions are important for further in vitro investigations and provide more relevant results than static culture conditions.

Density-dependence of functional spiking networks in vitro

Energy Technology Data Exchange (ETDEWEB)

Ham, Michael I [Los Alamos National Laboratory; Gintautuas, Vadas [Los Alamos National Laboratory; Rodriguez, Marko A [Los Alamos National Laboratory; Bettencourt, Luis M A [Los Alamos National Laboratory; Bennett, Ryan [UNIV OF NORTH TEXAS; Santa Maria, Cara L [UNIV OF NORTH TEXAS

2008-01-01

During development, the mammalian brain differentiates into specialized regions with unique functional abilities. While many factors contribute to this functional specialization, we explore the effect neuronal density can have on neuronal interactions. Two types of networks, dense (50,000 neurons and glia support cells) and sparse (12,000 neurons and glia support cells), are studied. A competitive first response model is applied to construct activation graphs that represent pairwise neuronal interactions. By observing the evolution of these graphs during development in vitro we observe that dense networks form activation connections earlier than sparse networks, and that link-!llltropy analysis of the resulting dense activation graphs reveals that balanced directional connections dominate. Information theoretic measures reveal in addition that early functional information interactions (of order 3) are synergetic in both dense and sparse networks. However, during development in vitro, such interactions become redundant in dense, but not sparse networks. Large values of activation graph link-entropy correlate strongly with redundant ensembles observed in the dense networks. Results demonstrate differences between dense and sparse networks in terms of informational groups, pairwise relationships, and activation graphs. These differences suggest that variations in cell density may result in different functional specialization of nervous system tissue also in vivo.

Antifungal Activity of Colistin against Mucorales Species In Vitro and in a Murine Model of Rhizopus oryzae Pulmonary Infection▿

Science.gov (United States)

Ben-Ami, Ronen; Lewis, Russell E.; Tarrand, Jeffrey; Leventakos, Konstantinos; Kontoyiannis, Dimitrios P.

2010-01-01

In immunosuppressed hosts, mucormycosis is a life-threatening infection with few treatment options. We studied the activity of colistin (polymyxin E) against Mucorales species in vitro and in a murine model of pulmonary Rhizopus oryzae infection. Colistin exhibited fungicidal activity in vitro against Mucorales spores and mycelia. At the colistin MIC, initial R. oryzae hyphal damage was followed by rapid regrowth; however, regrowth was prevented by combining colistin with a subinhibitory concentration of amphotericin B. Using electron microscopy and FM4-64 staining, we demonstrated that colistin disrupts R. oryzae cytoplasmic and vacuolar membranes, resulting in the leakage of intracellular contents. The prophylactic intranasal treatment of immunosuppressed mice with colistimethate significantly reduced the mortality rate and pulmonary fungal burden resulting from inhalational challenge with R. oryzae spores, whereas intraperitoneal colistimethate treatment had no effect. We conclude that colistin has modest in vitro and in vivo fungicidal activity against Mucorales spp. Further studies are warranted to assess the use of this drug in the prevention and treatment of mucormycosis. PMID:19858263

Antifungal activity of colistin against mucorales species in vitro and in a murine model of Rhizopus oryzae pulmonary infection.

Science.gov (United States)

Ben-Ami, Ronen; Lewis, Russell E; Tarrand, Jeffrey; Leventakos, Konstantinos; Kontoyiannis, Dimitrios P

2010-01-01

In immunosuppressed hosts, mucormycosis is a life-threatening infection with few treatment options. We studied the activity of colistin (polymyxin E) against Mucorales species in vitro and in a murine model of pulmonary Rhizopus oryzae infection. Colistin exhibited fungicidal activity in vitro against Mucorales spores and mycelia. At the colistin MIC, initial R. oryzae hyphal damage was followed by rapid regrowth; however, regrowth was prevented by combining colistin with a subinhibitory concentration of amphotericin B. Using electron microscopy and FM4-64 staining, we demonstrated that colistin disrupts R. oryzae cytoplasmic and vacuolar membranes, resulting in the leakage of intracellular contents. The prophylactic intranasal treatment of immunosuppressed mice with colistimethate significantly reduced the mortality rate and pulmonary fungal burden resulting from inhalational challenge with R. oryzae spores, whereas intraperitoneal colistimethate treatment had no effect. We conclude that colistin has modest in vitro and in vivo fungicidal activity against Mucorales spp. Further studies are warranted to assess the use of this drug in the prevention and treatment of mucormycosis.

Evaluation of the success rate of cone beam computed tomography in determining the location and direction of screw access holes in cement-retained implant-supported prostheses: An in vitro study.

Science.gov (United States)

Neshandar Asli, Hamid; Dalili Kajan, Zahra; Gholizade, Fatemeh

2018-02-21

Cement-retained implant-supported restorations have advantages over screw-retained restorations but are difficult to retrieve. Identifying the approximate location of the screw access hole (SAH) may reduce damage to the prosthesis. The purpose of this in vitro study was to evaluate the ability of cone beam computed tomography (CBCT) imaging to determine the location and direction of SAHs in cement-retained implant prostheses. Five clear acrylic resin casts were made based on a mandibular model. Several implant osteotomies (n=30) were created on the models with surgical burs, and crowns were made using the standard laboratory method with a transfer coping and the closed tray impression technique. CBCT images from the acrylic resin casts were evaluated by a maxillofacial radiologist who was blind to the locations and angles of the osteotomies. The locations of the access holes were determined on multiplanar reconstruction images and transferred to the clinical crown surface as defined points. Based on cross-sectional images, the predicted angle of the access hole was provided to a prosthodontist who was requested to pierce the crown at the proposed location in the specified direction. If the location and/or direction of the access hole were found, the process was considered successful, as the crown could then be removed from the implant abutment through the SAH. The success rate in the detection of the location and direction of the SAH was calculated, and chi-square and Fisher exact tests were applied for data analysis (α=.05). According to the results of this study, the success rate of CBCT to define the location of SAHs was 83.3% and 80% to determine the direction. No significant differences were found among the different dental groups in determination of the location (P=.79) or the direction (P=.53) of the SAHs. Most of the failures in determining the location and direction of the access hole in the buccolingual and mesiodistal directions were in the buccal and

Rethinking the globalisation of problem-based learning: how culture challenges self-directed learning.

NARCIS (Netherlands)

Frambach, J.M.; Driessen, E.W.; Chan, L.C.; Vleuten, C.P.M. van der

2012-01-01

Medical Education 2012: 46: 738-747 Context Medical schools worldwide are increasingly switching to student-centred methods such as problem-based learning (PBL) to foster lifelong self-directed learning (SDL). The cross-cultural applicability of these methods has been questioned because of their

New European BSS - Challenges And Solutions

International Nuclear Information System (INIS)

Janzekovic, H.

2015-01-01

Today the European Basic Safety Standards (2014) is the central legal document in the Europe Union focusing on harmonization of radiation protection in Member States (MSs). It forms, together with Nuclear Safety Directive and Spent Fuel and Radioactive Waste Directive, a pillar of a comprehensive network of rules. The BSS Directive is, as the IAEA BSS Directive (2014), based on the three exposure situations recommended in the ICRP 103. This Directive is a complex and in some part very technical document taking into account development of science, regulatory practices from the past and the emerging needs of Member States. A list of its new provisions is very long. One of the main cornerstones of the text is enhanced collaboration among MSs as well as among MSs and the European Commission. The BSS Directive envisages the enhancement in three areas, i.e. in control of consumer products, recognition of experts and services as well as in activities related to emergency exposure situations. Four additional identified areas where such collaboration might be useful, pose challenges to MSs, especially to neighbouring countries with strong connections. They are related to regulatory regime including clearance regime and control of NORM, workers, building materials and lessons learned from accidents and other events related to radiation. Further guidance on these issues would be beneficial. (author).

The immunoglobulin class of anti-hapten antibody secreted during secondary responses in vitro and in vivo.

Science.gov (United States)

North, J R; Dresser, D W

1977-05-01

A comparison has been made of the in vitro and in vivo response of primed mouse spleen cells to the hapten DNP. The responses were analysed in terms of six classes (sub-classes) of humoral antibody directed against the cross-reacting hapten TNP. By comparison with the response in intact mice the adoptive secondary response is delayed by 3 days in addition to being somewhat lesser in magnitude. The timing of the response in vitro is similar to that observed in intact mice. The preponderant class in all three responses was gammaG1 with gammaA and gammaG3 secreting cells consistently comprising the smallest proportion of the total of antibody-secreting cells.

In vitro eugenics.

Science.gov (United States)

Sparrow, Robert

2014-11-01

A series of recent scientific results suggest that, in the not-too-distant future, it will be possible to create viable human gametes from human stem cells. This paper discusses the potential of this technology to make possible what I call 'in vitro eugenics': the deliberate breeding of human beings in vitro by fusing sperm and egg derived from different stem-cell lines to create an embryo and then deriving new gametes from stem cells derived from that embryo. Repeated iterations of this process would allow scientists to proceed through multiple human generations in the laboratory. In vitro eugenics might be used to study the heredity of genetic disorders and to produce cell lines of a desired character for medical applications. More controversially, it might also function as a powerful technology of 'human enhancement' by allowing researchers to use all the techniques of selective breeding to produce individuals with a desired genotype. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Facing Tomorrow's Challenges - An Overview

Science.gov (United States)

,

2008-01-01

In 2007, the U.S. Geological Survey (USGS) developed a science strategy outlining the major natural-science issues facing the Nation in the next decade. The science strategy consists of six science directions of critical importance, focusing on areas where natural science can make a substantial contribution to the well-being of the Nation and the world. This fact sheet is an overview of the science strategy and describes how USGS research can strengthen the Nation with information needed to meet the challenges of the 21st century.

Reproductive effects of two neonicotinoid insecticides on mouse sperm function and early embryonic development in vitro.

Directory of Open Access Journals (Sweden)

Yi-Hua Gu

Full Text Available Acetamiprid (ACE and imidacloprid (IMI are two major members in the family of neonicotinoid pesticides, which are synthesized with a higher selectivity to insects. The present study determined and compared in vitro effects of ACE, IMI and nicotine on mammalian reproduction by using an integrated testing strategy for reproductive toxicology, which covered sperm quality, sperm penetration into oocytes and preimplantation embryonic development. Direct chemical exposure (500 µM or 5 mM on spermatozoa during capacitation was performed, and in vitro fertilization (IVF process, zygotes and 2-cell embryos were respectively incubated with chemical-supplemented medium until blastocyst formation to evaluate the reproductive toxicity of these chemicals and monitor the stages mainly affected. Generally, treatment of 500 µM or 5 mM chemicals for 30 min did not change sperm motility and DNA integrity significantly but the fertilization ability in in vitro fertilization (IVF process, indicating that IVF process could detect and distinguish subtle effect of spermatozoa exposed to different chemicals. Culture experiment in the presence of chemicals in medium showed that fertilization process and zygotes are adversely affected by direct exposure of chemicals (PIMI>ACE, whereas developmental progression of 2-cell stage embryos was similar to controls (P>0.05. These findings unveiled the hazardous effects of neonicotinoid pesticides exposure on mammalian sperm fertilization ability as well as embryonic development, raising the concerns that neonicotinoid pesticides may pose reproductive risks on human reproductive health, especially in professional populations.

Escherichia coli Uropathogenesis In Vitro

DEFF Research Database (Denmark)

Andersen, Thomas E; Khandige, Surabhi; Madelung, Michelle

2012-01-01

-stage infection events have not been replicated in vitro. We have established an in vitro model of human bladder cell infection by the use of a flow chamber (FC)-based culture system, which allows investigation of steps subsequent to initial invasion. Short-term bacterial colonization on the FC-BEC layer led...... to rods that could invade other BECs. Hence, under growth conditions established to resemble those present in vivo, the elements of the proposed uropathogenic cascade were inducible in a human BEC model system. Here, we describe the model and show how these characteristics are reproduced in vitro....

In vitro toxicity and control of Meloidogyne incognita in soybean by rosemary extract

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Mônica Anghinoni Müller

2016-02-01

Full Text Available The control of nematodes in plants can be challenging, and there is a need for alternative, environmentally conscious methods for their management. The purpose of this study was to evaluate the effect of rosemary extract (Rosmarinus officinalis on the in vitro toxicity and control of Meloidogyne incognita in CD 206 and CD 215 soybean cultivars. Using an in vitro assay, 500 M. incognita eggs per plate were observed for 15 days after incubation with rosemary extract at concentrations of 1%, 5%, and 10%. Soybean plants were studied under greenhouse conditions, and starting at V3 stage, were sprayed weekly with the same concentration of rosemary extract for 64 days. Three days after the first treatment, each soybean plant was inoculated with 1800 eggs and 400 second-stage juveniles (J2. At the end of this essay, number of eggs and J2 in the roots and soil, number of galls, and the reproduction factor (RF were evaluated. Our results showed that in the in vitro assay, rosemary extract reduced the number of M. incognita eggs that hatched. Under greenhouse conditions, the CD 206 cultivar showed a 48% reduction in the number of galls, as well as fewer eggs in the soil and a lower RF. Similarly, in the CD 215 cultivar, the number of eggs was reduced and the RF was lower. These results indicate the potential for rosemary extract to control M. incognita in soybean crops.

Validation of an immortalized human (hBMEC) in vitro blood-brain barrier model.

Science.gov (United States)

Eigenmann, Daniela Elisabeth; Jähne, Evelyn Andrea; Smieško, Martin; Hamburger, Matthias; Oufir, Mouhssin

2016-03-01

We recently established and optimized an immortalized human in vitro blood-brain barrier (BBB) model based on the hBMEC cell line. In the present work, we validated this mono-culture 24-well model with a representative series of drug substances which are known to cross or not to cross the BBB. For each individual compound, a quantitative UHPLC-MS/MS method in Ringer HEPES buffer was developed and validated according to current regulatory guidelines, with respect to selectivity, precision, and reliability. Various biological and analytical challenges were met during method validation, highlighting the importance of careful method development. The positive controls antipyrine, caffeine, diazepam, and propranolol showed mean endothelial permeability coefficients (P e) in the range of 17-70 × 10(-6) cm/s, indicating moderate to high BBB permeability when compared to the barrier integrity marker sodium fluorescein (mean P e 3-5 × 10(-6) cm/s). The negative controls atenolol, cimetidine, and vinblastine showed mean P e values < 10 × 10(-6) cm/s, suggesting low permeability. In silico calculations were in agreement with in vitro data. With the exception of quinidine (P-glycoprotein inhibitor and substrate), BBB permeability of all control compounds was correctly predicted by this new, easy, and fast to set up human in vitro BBB model. Addition of retinoic acid and puromycin did not increase transendothelial electrical resistance (TEER) values of the BBB model.

Tuning cell adhesion by direct nanostructuring silicon into cell repulsive/adhesive patterns

International Nuclear Information System (INIS)

Premnath, Priyatha; Tavangar, Amirhossein; Tan, Bo; Venkatakrishnan, Krishnan

2015-01-01

Developing platforms that allow tuning cell functionality through incorporating physical, chemical, or mechanical cues onto the material surfaces is one of the key challenges in research in the field of biomaterials. In this respect, various approaches have been proposed and numerous structures have been developed on a variety of materials. Most of these approaches, however, demand a multistep process or post-chemical treatment. Therefore, a simple approach would be desirable to develop bio-functionalized platforms for effectively modulating cell adhesion and consequently programming cell functionality without requiring any chemical or biological surface treatment. This study introduces a versatile yet simple laser approach to structure silicon (Si) chips into cytophobic/cytophilic patterns in order to modulate cell adhesion and proliferation. These patterns are fabricated on platforms through direct laser processing of Si substrates, which renders a desired computer-generated configuration into patterns. We investigate the morphology, chemistry, and wettability of the platform surfaces. Subsequently, we study the functionality of the fabricated platforms on modulating cervical cancer cells (HeLa) behaviour. The results from in vitro studies suggest that the nanostructures efficiently repel HeLa cells and drive them to migrate onto untreated sites. The study of the morphology of the cells reveals that cells evade the cytophobic area by bending and changing direction. Additionally, cell patterning, cell directionality, cell channelling, and cell trapping are achieved by developing different platforms with specific patterns. The flexibility and controllability of this approach to effectively structure Si substrates to cell-repulsive and cell-adhesive patterns offer perceptible outlook for developing bio-functionalized platforms for a variety of biomedical devices. Moreover, this approach could pave the way for developing anti-cancer platforms that selectively repel

Tuning cell adhesion by direct nanostructuring silicon into cell repulsive/adhesive patterns

Energy Technology Data Exchange (ETDEWEB)

Premnath, Priyatha, E-mail: priyatha.premnath@ryerson.ca [Micro/Nanofabrication Laboratory, Department of Mechanical and Industrial Engineering, Ryerson University, 350 Victoria Street, Toronto, ON, Canada M5B 2K3 (Canada); Tavangar, Amirhossein, E-mail: atavanga@ryerson.ca [Micro/Nanofabrication Laboratory, Department of Mechanical and Industrial Engineering, Ryerson University, 350 Victoria Street, Toronto, ON, Canada M5B 2K3 (Canada); Tan, Bo, E-mail: tanbo@ryerson.ca [Nanocharacterization Laboratory, Department of Aerospace Engineering, Ryerson University, 350 Victoria Street, Toronto, ON, Canada M5B 2K3 (Canada); Venkatakrishnan, Krishnan, E-mail: venkat@ryerson.ca [Micro/Nanofabrication Laboratory, Department of Mechanical and Industrial Engineering, Ryerson University, 350 Victoria Street, Toronto, ON, Canada M5B 2K3 (Canada)

2015-09-10

Developing platforms that allow tuning cell functionality through incorporating physical, chemical, or mechanical cues onto the material surfaces is one of the key challenges in research in the field of biomaterials. In this respect, various approaches have been proposed and numerous structures have been developed on a variety of materials. Most of these approaches, however, demand a multistep process or post-chemical treatment. Therefore, a simple approach would be desirable to develop bio-functionalized platforms for effectively modulating cell adhesion and consequently programming cell functionality without requiring any chemical or biological surface treatment. This study introduces a versatile yet simple laser approach to structure silicon (Si) chips into cytophobic/cytophilic patterns in order to modulate cell adhesion and proliferation. These patterns are fabricated on platforms through direct laser processing of Si substrates, which renders a desired computer-generated configuration into patterns. We investigate the morphology, chemistry, and wettability of the platform surfaces. Subsequently, we study the functionality of the fabricated platforms on modulating cervical cancer cells (HeLa) behaviour. The results from in vitro studies suggest that the nanostructures efficiently repel HeLa cells and drive them to migrate onto untreated sites. The study of the morphology of the cells reveals that cells evade the cytophobic area by bending and changing direction. Additionally, cell patterning, cell directionality, cell channelling, and cell trapping are achieved by developing different platforms with specific patterns. The flexibility and controllability of this approach to effectively structure Si substrates to cell-repulsive and cell-adhesive patterns offer perceptible outlook for developing bio-functionalized platforms for a variety of biomedical devices. Moreover, this approach could pave the way for developing anti-cancer platforms that selectively repel

The Influence of Staff Training on Challenging Behaviour in Individuals with Intellectual Disability: A Review

Science.gov (United States)

Cox, Alison D.; Dube, Charmayne; Temple, Beverley

2015-01-01

Many individuals with intellectual disability engage in challenging behaviour. This can significantly limit quality of life and also negatively impact caregivers (e.g., direct care staff, family caregivers and teachers). Fortunately, efficacious staff training may alleviate some negative side effects of client challenging behaviour. Currently, a…

Directed PCR-free engineering of highly repetitive DNA sequences

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Preissler Steffen

2011-09-01

Full Text Available Abstract Background Highly repetitive nucleotide sequences are commonly found in nature e.g. in telomeres, microsatellite DNA, polyadenine (poly(A tails of eukaryotic messenger RNA as well as in several inherited human disorders linked to trinucleotide repeat expansions in the genome. Therefore, studying repetitive sequences is of biological, biotechnological and medical relevance. However, cloning of such repetitive DNA sequences is challenging because specific PCR-based amplification is hampered by the lack of unique primer binding sites resulting in unspecific products. Results For the PCR-free generation of repetitive DNA sequences we used antiparallel oligonucleotides flanked by restriction sites of Type IIS endonucleases. The arrangement of recognition sites allowed for stepwise and seamless elongation of repetitive sequences. This facilitated the assembly of repetitive DNA segments and open reading frames encoding polypeptides with periodic amino acid sequences of any desired length. By this strategy we cloned a series of polyglutamine encoding sequences as well as highly repetitive polyadenine tracts. Such repetitive sequences can be used for diverse biotechnological applications. As an example, the polyglutamine sequences were expressed as His6-SUMO fusion proteins in Escherichia coli cells to study their aggregation behavior in vitro. The His6-SUMO moiety enabled affinity purification of the polyglutamine proteins, increased their solubility, and allowed controlled induction of the aggregation process. We successfully purified the fusions proteins and provide an example for their applicability in filter retardation assays. Conclusion Our seamless cloning strategy is PCR-free and allows the directed and efficient generation of highly repetitive DNA sequences of defined lengths by simple standard cloning procedures.

The harmonized INFOGEST in vitro digestion method

NARCIS (Netherlands)

Egger, Lotti; Ménard, Olivia; Delgado-Andrade, Cristina; Alvito, Paula; Assunção, Ricardo; Balance, Simon; Barberá, Reyes; Brodkorb, Andre; Cattenoz, Thomas; Clemente, Alfonso; Comi, Irene; Dupont, Didier; Garcia-Llatas, Guadalupe; Lagarda, María Jesús; Feunteun, Le Steven; Janssen Duijghuijsen, Lonneke; Karakaya, Sibel; Lesmes, Uri; Mackie, Alan R.; Martins, Carla; Meynier, Anne; Miralles, Beatriz; Murray, B.S.; Pihlanto, Anne; Picariello, Gianluca; Santos, C.N.; Simsek, Sebnem; Recio, Isidra; Rigby, Neil; Rioux, Laurie Eve; Stoffers, Helena; Tavares, Ana; Tavares, Lucelia; Turgeon, Sylvie; Ulleberg, E.K.; Vegarud, G.E.; Vergères, Guy; Portmann, Reto

2016-01-01

Within the active field of in vitro digestion in food research, the COST Action INFOGEST aimed to harmonize in vitro protocols simulating human digestion on the basis of physiologically inferred conditions. A harmonized static in vitro digestion (IVD) method was recently published as a primary

[Ambulatory pediatrics: a challenge].

Science.gov (United States)

Ransy, V; Gevers, B; Landsberg, M

2006-01-01

Ambulatory paediatrics in University hospitals has remarkably evolved during the past decade, along with technological progress and the current need for undelayed information and attention; demand for hospital medical advice increases consequently, either directly in outpatients wards or indirectly by phone or e-mails. Specific medico-social aspects linked essentially to populations' migration, poverty, chronic stress and family splitting are regularly encountered. Hospital architecture and adequacy of medical and nursing staff must both be adjusted to these changing medical demands including medical teaching. We now face the ever-growing challenge of providing an adequate management of actual medico-psycho-social aspects and integrating up-to-date paediatrics in our daily practices.

Utility of tantalum (Ta) coating to improve surface hardness in vitro bioactivity and biocompatibility of Co–Cr

Energy Technology Data Exchange (ETDEWEB)

Pham, Vuong-Hung [WCU Hybrid Materials Program, Department of Materials Science and Engineering, Seoul National University, Seoul, 151-742 (Korea, Republic of); Advanced Institute for Science and Technology (AIST), Hanoi University of Science and Technology (HUST), No 1, Dai Co Viet Road, Ha Noi (Viet Nam); Lee, Seung-Hee; Li, Yuanlong; Kim, Hyoun-Ee [WCU Hybrid Materials Program, Department of Materials Science and Engineering, Seoul National University, Seoul, 151-742 (Korea, Republic of); Shin, Kwan-Ha [Department of Dental Laboratory Science and Engineering, Korea University, Seoul, 136-703 (Korea, Republic of); Koh, Young-Hag, E-mail: kohyh@korea.ac.kr [Department of Dental Laboratory Science and Engineering, Korea University, Seoul, 136-703 (Korea, Republic of)

2013-06-01

This study reports the utility of tantalum (Ta) coating for improving the surface hardness, in vitro bioactivity and biocompatibility of Co–Cr implants. The use of direct current sputtering allowed for the deposition of a dense and uniform Ta film onto a Co–Cr substrate, which was composed of β-phase Ta grains. This hard Ta coating significantly improved the surface hardness of the Co–Cr by a factor of > 2.3. In addition, the Ta-deposited Co–Cr substrate showed a vigorous precipitation of apatite crystals on its surface after 4 weeks of immersion in simulated body fluid, suggesting its excellent in vitro bioactivity. This bioactive Ta coating led to a considerable improvement in the in vitro biocompatibility of the Co–Cr, which was assessed in terms of the attachment, proliferation and differentiation of pre-osteoblasts (MC3T3-E1). - Highlights: • Dense and uniform Ta film was deposited onto a Co–Cr substrate using DC sputtering. • The Ta coating significantly enhanced the surface hardness of the Co–Cr. • The in vitro biocompatibility of the Co–Cr was also significantly improved.

Utility of tantalum (Ta) coating to improve surface hardness in vitro bioactivity and biocompatibility of Co–Cr

International Nuclear Information System (INIS)

Pham, Vuong-Hung; Lee, Seung-Hee; Li, Yuanlong; Kim, Hyoun-Ee; Shin, Kwan-Ha; Koh, Young-Hag

2013-01-01

This study reports the utility of tantalum (Ta) coating for improving the surface hardness, in vitro bioactivity and biocompatibility of Co–Cr implants. The use of direct current sputtering allowed for the deposition of a dense and uniform Ta film onto a Co–Cr substrate, which was composed of β-phase Ta grains. This hard Ta coating significantly improved the surface hardness of the Co–Cr by a factor of > 2.3. In addition, the Ta-deposited Co–Cr substrate showed a vigorous precipitation of apatite crystals on its surface after 4 weeks of immersion in simulated body fluid, suggesting its excellent in vitro bioactivity. This bioactive Ta coating led to a considerable improvement in the in vitro biocompatibility of the Co–Cr, which was assessed in terms of the attachment, proliferation and differentiation of pre-osteoblasts (MC3T3-E1). - Highlights: • Dense and uniform Ta film was deposited onto a Co–Cr substrate using DC sputtering. • The Ta coating significantly enhanced the surface hardness of the Co–Cr. • The in vitro biocompatibility of the Co–Cr was also significantly improved

Identification of Arsenic Direct-Binding Proteins in Acute Promyelocytic Leukaemia Cells

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Tao Zhang

2015-11-01

Full Text Available The identification of arsenic direct-binding proteins is essential for determining the mechanism by which arsenic trioxide achieves its chemotherapeutic effects. At least two cysteines close together in the amino acid sequence are crucial to the binding of arsenic and essential to the identification of arsenic-binding proteins. In the present study, arsenic binding proteins were pulled down with streptavidin and identified using a liquid chromatograph-mass spectrometer (LC-MS/MS. More than 40 arsenic-binding proteins were separated, and redox-related proteins, glutathione S-transferase P1 (GSTP1, heat shock 70 kDa protein 9 (HSPA9 and pyruvate kinase M2 (PKM2, were further studied using binding assays in vitro. Notably, PKM2 has a high affinity for arsenic. In contrast to PKM2, GSTP1and HSPA9 did not combine with arsenic directly in vitro. These observations suggest that arsenic-mediated acute promyelocytic leukaemia (APL suppressive effects involve PKM2. In summary, we identified several arsenic binding proteins in APL cells and investigated the therapeutic mechanisms of arsenic trioxide for APL. Further investigation into specific signal pathways by which PKM2 mediates APL developments may lead to a better understanding of arsenic effects on APL.