Micropropagation of Asparagus by in vitro shoot culture.

PubMed

Stajner, Nataša

2013-01-01

Asparagus officinalis is most extensively studied species within the genus Asparagus, which is well known as garden asparagus. This species is dioecious with unisexual flowers, which means that generative propagation gives roughly equal number of male and female plants. Male plants are high yielders and preferred commercially over female plants. Tissue culture techniques could efficiently promote vegetative propagation of male plants and pave the way for efficient plant breeding.This chapter describes an efficient micropropagation protocol for developing rapid growing in vitro Asparagus shoot cultures. The source of explants, inoculation, and shoot proliferation, followed by shoot propagation, rooting, and acclimatization is described. The optimal medium for Asparagus micropropagation described in this chapter is composed of MS macro- and microelements and a combination of auxins and cytokinins. Plant growth regulators NAA, kinetin, and BA were used in various concentrations. Three different media representing the whole micropropagation protocol of Asparagus are described; medium for shoot initiation, medium for shoot multiplication, and medium for root formation. By in vitro propagation of Asparagus, root initiation is difficult, but can be promoted by adding growth retardant ancymidol which also greatly promotes shoot development and suppresses callus formation.

Preparation and Fluorescent Analysis of Plant Metaphase Chromosomes.

PubMed

Schwarzacher, Trude

2016-01-01

Good preparations are essential for informative analysis of both somatic and meiotic chromosomes, cytogenetics, and cell divisions. Fluorescent chromosome staining allows even small chromosomes to be visualized and counted, showing their morphology. Aneuploidies and polyploidies can be established for species, populations, or individuals while changes occurring in breeding lines during hybridization or tissue culture and transformation protocols can be assessed. The process of division can be followed during mitosis and meiosis including pairing and chiasma distribution, as well as DNA organization and structure during the evolution of chromosomes can be studied. This chapter presents protocols for pretreatment and fixation of material, including tips of how to grow plants to get good and healthy meristem with many divisions. The chromosome preparation technique is described using proteolytic enzymes, but acids can be used instead. Chromosome slide preparations are suitable for fluorochrome staining for fast screening (described in the chapter) or fluorescent in situ hybridization (see Schwarzacher and Heslop-Harrison, In situ hybridization. BIOS Scientific Publishers, Oxford, 2000).

Study and development of label-free optical biosensors for biomedical applications

NASA Astrophysics Data System (ADS)

Choi, Charles J.

For the majority of assays currently performed, fluorescent or colorimetric chemical labels are commonly attached to the molecules under study so that they may be readily visualized. The methods of using labels to track biomolecular binding events are very sensitive and effective, and are employed as standardized assay protocol across research labs worldwide. However, using labels induces experimental uncertainties due to the effect of the label on molecular conformation, active binding sites, or inability to find an appropriate label that functions equivalently for all molecules in an experiment. Therefore, the ability to perform highly sensitive biochemical detection without the use of fluorescent labels would further simplify assay protocols and would provide quantitative kinetic data, while removing experimental artifacts from fluorescent quenching, shelf-life, and background fluorescence phenomena. In view of the advantages mentioned above, the study and development of optical label-free sensor technologies have been undertaken here. In general, label-free photonic crystal (PC) biosensors and metal nanodome array surface-enhanced Raman scattering (SERS) substrates, both of which are fabricated by nanoreplica molding process, have been used as the method to attack the problem. Chapter 1 shows the work on PC label-free biosensor incorporated microfluidic network for bioassay performance enhancement and kinetic reaction rate constant determination. Chapter 2 describes the work on theoretical and experimental comparison of label-free biosensing in microplate, microfluidic, and spot-based affinity capture assays. Chapter 3 shows the work on integration of PC biosensor with actuate-to-open valve microfluidic chip for pL-volume combinatorial mixing and screening application. In Chapter 4, the development and characterization of SERS nanodome array is shown. Lastly, Chapter 5 describes SERS nanodome sensor incorporated tubing for point-of-care monitoring of intravenous drugs and metabolites.

Modeling Ebola Virus Genome Replication and Transcription with Minigenome Systems.

PubMed

Cressey, Tessa; Brauburger, Kristina; Mühlberger, Elke

2017-01-01

In this chapter, we describe the minigenome system for Ebola virus (EBOV), which reconstitutes EBOV polymerase activity in cells and can be used to model viral genome replication and transcription. This protocol comprises all steps including cell culture, plasmid preparation, transfection, and luciferase reporter assay readout.

Equipment and Protocols for Quasi-Static and Dynamic Tests of Very-High-Strength Concrete (VHSC) and High-Strength High-Ductility Concrete (HSHDC)

DTIC Science & Technology

2016-08-01

quasi -static mechanical properties, deformation behavior, and damage mechanisms in HSHDC and compare the behavior with VHSC. 2. Develop experimental ...using the experimental setup described in Chapter 6. The quasi -static strain rate was approximately 10-4/s. All panels tested have nominal dimensions...ER D C TR -1 6- 13 Force Protection Basing; TeCD 1a Equipment and Protocols for Quasi -Static and Dynamic Tests of Very-High-Strength

Anther Culture in Pepper (Capsicum annuum L.).

PubMed

Parra-Vega, Verónica; Seguí-Simarro, Jose M

2016-01-01

Anther culture is the most popular of the techniques used to induce microspore embryogenesis. This technique is well set up in a wide range of crops, including pepper. In this chapter, a protocol for anther culture in pepper is described. The protocol presented hereby includes the steps from the selection of buds from donor plants to the regeneration and acclimatization of doubled haploid plants derived from the embryos, as well as a description of how to analyze the ploidy level of the regenerated plants.

Chapter A5. Processing of Water Samples

USGS Publications Warehouse

Wilde, Franceska D.; Radtke, Dean B.; Gibs, Jacob; Iwatsubo, Rick T.

1999-01-01

The National Field Manual for the Collection of Water-Quality Data (National Field Manual) describes protocols and provides guidelines for U.S. Geological Survey (USGS) personnel who collect data used to assess the quality of the Nation's surface-water and ground-water resources. This chapter addresses methods to be used in processing water samples to be analyzed for inorganic and organic chemical substances, including the bottling of composite, pumped, and bailed samples and subsamples; sample filtration; solid-phase extraction for pesticide analyses; sample preservation; and sample handling and shipping. Each chapter of the National Field Manual is published separately and revised periodically. Newly published and revised chapters will be announced on the USGS Home Page on the World Wide Web under 'New Publications of the U.S. Geological Survey.' The URL for this page is http:/ /water.usgs.gov/lookup/get?newpubs.

Analysis of the Cellular Stress Response During Ebola Virus Infection by Immunofluorescence.

PubMed

Nelson, Emily V; Schmidt, Kristina M

2017-01-01

In this chapter, the use of immunofluorescence analysis as a tool to examine stress granule (SG) formation in Ebola virus (EBOV)-infected cells is described. The following protocol focuses on the process of inducing and analyzing the cellular stress response, including treatment of cells with inducers and inhibitors of the SG formation, and also describes EBOV infection, DNA transfection, and the usage of different cell lines.

A Teacher's Project Guide to the Internet.

ERIC Educational Resources Information Center

Crotchett, Kevin R.

This book is a step-by-step guide to the Internet, suggesting creative K-12 classroom projects ranging from one of the most basic Internet functions, e-mail, to one of the more difficult, writing World Wide Web homepages. Intervening chapters describe projects using Usenet Newsgroups, File Transfer Protocols (FTP), Gopher, Veronica, the World Wide…

106-17 Telemetry Management Resources Chapter 25

DTIC Science & Technology

2017-07-01

aspects of the TmNS system . There are two primary protocols for accessing the management resources: Simple Network Management Protocol (SNMP) and... management resources as well as a basic HTTP clients and servers for a more RESTful approach to system management . Both tools are available from the...Telemetry Standards, RCC Standard 106-17 Chapter 25, July 2017 i CHAPTER 25 Management Resources Acronyms

Preparation of gold nanocluster bioconjugates for electron microscopy.

PubMed

Heinecke, Christine L; Ackerson, Christopher J

2013-01-01

In this chapter, we describe types of gold nanoparticle-biomolecule conjugates and their use in electron microscopy. Included are two detailed protocols for labeling an IgG antibody with gold monolayer protected clusters. The first approach is a direct bonding approach that utilizes the ligand place exchange reaction. The second approach describes NHS-EDC coupling of Au(144)(pMBA)(60) with IgG. Also included are various characterization techniques for determining labeling efficiency.

Chapter A9. Safety in Field Activities

USGS Publications Warehouse

Lane, Susan L.; Ray, Ronald G.

1998-01-01

The National Field Manual for the Collection of Water-Quality Data (National Field Manual) describes protocols (requirements and recommendations) and provides guidelines for U.S. Geological Survey (USGS) personnel who collect data used to assess the quality of the Nation's surface-water and ground-water resources. This chapter of the manual addresses topics related to personal safety to be used in the collection of water-quality data, including: policies and general regulations on field safety; transportation of people and equipment; implementation of surface-water and ground-water activities; procedures for handling chemicals; and information on potentially hazardous environmental conditions, animals, and plants. Each chapter of the National Field Manual is published separately and revised periodically. Newly published and revised chapters will be announced on the USGS Home Page on the World Wide Web under 'New Publications of the U.S. Geological Survey.' The URL for this page is http://pubs.usgs.gov/publications/ index.html.

Protein blotting protocol for beginners.

PubMed

Petrasovits, Lars A

2014-01-01

The transfer and immobilization of biological macromolecules onto solid nitrocellulose or nylon (polyvinylidene difluoride (PVDF)) membranes subsequently followed by specific detection is referred to as blotting. DNA blots are called Southerns after the inventor of the technique, Edwin Southern. By analogy, RNA blots are referred to as northerns and protein blots as westerns (Burnette, Anal Biochem 112:195-203, 1981). With few exceptions, western blotting involves five steps, namely, sample collection, preparation, separation, immobilization, and detection. In this chapter, protocols for the entire process from sample collection to detection are described.

Tools for Model Building and Optimization into Near-Atomic Resolution Electron Cryo-Microscopy Density Maps.

PubMed

DiMaio, F; Chiu, W

2016-01-01

Electron cryo-microscopy (cryoEM) has advanced dramatically to become a viable tool for high-resolution structural biology research. The ultimate outcome of a cryoEM study is an atomic model of a macromolecule or its complex with interacting partners. This chapter describes a variety of algorithms and software to build a de novo model based on the cryoEM 3D density map, to optimize the model with the best stereochemistry restraints and finally to validate the model with proper protocols. The full process of atomic structure determination from a cryoEM map is described. The tools outlined in this chapter should prove extremely valuable in revealing atomic interactions guided by cryoEM data. © 2016 Elsevier Inc. All rights reserved.

Finger millet [Eleusine coracana (L.) Gaertn].

PubMed

Ceasar, Stanislaus Antony; Ignacimuthu, Savarimuthu

2015-01-01

Millets are the primary food source for millions of people in tropical regions of the world supplying mineral nutrition and protein. In this chapter, we describe an optimized protocol for the Agrobacterium-mediated transformation of finger millet variety GPU 45. Agrobacterium strain LBA4404 harboring plasmid pCAMBIA1301 which contains hygromycin phosphotransferase (hph) as selectable marker gene and β-glucuronidase (GUS) as reporter gene has been used. This protocol utilizes the shoot apex explants for the somatic embryogenesis and regeneration of finger millet after the transformation by Agrobacterium. Desiccation of explants during cocultivation helps for the better recovery of transgenic plants. This protocol is very useful for the efficient production of transgenic plants in finger millet through Agrobacterium-mediated transformation.

Agents Based e-Commerce and Securing Exchanged Information

NASA Astrophysics Data System (ADS)

Al-Jaljouli, Raja; Abawajy, Jemal

Mobile agents have been implemented in e-Commerce to search and filter information of interest from electronic markets. When the information is very sensitive and critical, it is important to develop a novel security protocol that can efficiently protect the information from malicious tampering as well as unauthorized disclosure or at least detect any malicious act of intruders. In this chapter, we describe robust security techniques that ensure a sound security of information gathered throughout agent’s itinerary against various security attacks, as well as truncation attacks. A sound security protocol is described, which implements the various security techniques that would jointly prevent or at least detect any malicious act of intruders. We reason about the soundness of the protocol usingSymbolic Trace Analyzer (STA), a formal verification tool that is based on symbolic techniques. We analyze the protocol in key configurations and show that it is free of flaws. We also show that the protocol fulfils the various security requirements of exchanged information in MAS, including data-integrity, data-confidentiality, data-authenticity, origin confidentiality and data non-repudiability.

Methods to Assess Mitochondrial Morphology in Mammalian Cells Mounting Autophagic or Mitophagic Responses.

PubMed

Marchi, S; Bonora, M; Patergnani, S; Giorgi, C; Pinton, P

2017-01-01

It is widely acknowledged that mitochondria are highly active structures that rapidly respond to cellular and environmental perturbations by changing their shape, number, and distribution. Mitochondrial remodeling is a key component of diverse biological processes, ranging from cell cycle progression to autophagy. In this chapter, we describe different methodologies for the morphological study of the mitochondrial network. Instructions are given for the preparation of samples for fluorescent microscopy, based on genetically encoded strategies or the employment of synthetic fluorescent dyes. We also propose detailed protocols to analyze mitochondrial morphometric parameters from both three-dimensional and bidimensional datasets. Finally, we describe a protocol for the visualization and quantification of mitochondrial structures through electron microscopy. © 2017 Elsevier Inc. All rights reserved.

Potato (Solanum tuberosum L.).

PubMed

Chetty, Venkateswari J; Narváez-Vásquez, Javier; Orozco-Cárdenas, Martha L

2015-01-01

Agrobacterium-mediated transformation is the most common method for the incorporation of foreign genes into the genome of potato as well as many other species in the Solanaceae family. This chapter describes protocols for the genetic transformation of three species of potato: Solanum tuberosum subsp. tuberosum (Desiréé), S. tuberosum subsp. andigenum (Blue potato), and S. tuberosum subsp. andigena using internodal segments as explants.

Large-Scale Interaction Profiling of Protein Domains Through Proteomic Peptide-Phage Display Using Custom Peptidomes.

PubMed

Seo, Moon-Hyeong; Nim, Satra; Jeon, Jouhyun; Kim, Philip M

2017-01-01

Protein-protein interactions are essential to cellular functions and signaling pathways. We recently combined bioinformatics and custom oligonucleotide arrays to construct custom-made peptide-phage libraries for screening peptide-protein interactions, an approach we call proteomic peptide-phage display (ProP-PD). In this chapter, we describe protocols for phage display for the identification of natural peptide binders for a given protein. We finally describe deep sequencing for the analysis of the proteomic peptide-phage display.

Measuring Helicase Inhibition of the DEAD-box Protein Dbp2 by Yra1

PubMed Central

Ma, Wai Kit; Tran, Elizabeth J.

2016-01-01

Despite the highly conserved helicase core, individual DEAD-box proteins are specialized in diverse RNA metabolic processes. One mechanism that determines DEAD-box protein specificity is enzymatic regulation by other protein cofactors. In this chapter, we describe a protocol for purifying the Saccharomyces cerevisiae DEAD-box RNA helicase Dbp2 and RNA-binding protein Yra1 and subsequent analysis of helicase regulation. The experiments described here can be adapted to RNA helicase and purified co-factor. PMID:25579587

Chapter 10: Peak Demand and Time-Differentiated Energy Savings Cross-Cutting Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Stern, Frank; Spencer, Justin

Savings from electric energy efficiency measures and programs are often expressed in terms of annual energy and presented as kilowatt-hours per year (kWh/year). However, for a full assessment of the value of these savings, it is usually necessary to consider the measure or program's impact on peak demand as well as time-differentiated energy savings. This cross-cutting protocol describes methods for estimating the peak demand and time-differentiated energy impacts of measures implemented through energy efficiency programs.

106-17 Telemetry Standards Chapter 7 Packet Telemetry Downlink

DTIC Science & Technology

2017-07-31

Acronyms IP Internet Protocol IPv4 Internet Protocol, Version 4 IPv6 Internet Protocol, Version 6 LLP low-latency PTDP MAC media access control...o 4’b0101: PT Internet Protocol (IP) Packet o 4’b0110: PT Chapter 24 TmNSMessage Packet o 4’b0111 – 4’b1111: Reserved • Fragment (bits 17 – 16...packet is defined as a free -running 12-bit counter. The PT test counter packet shall consist of one 12-bit word and shall be encoded as one 24-bit

Micropatterning on micropost arrays.

PubMed

Sniadecki, Nathan J; Han, Sangyoon J; Ting, Lucas H; Feghhi, Shirin

2014-01-01

Micropatterning of cells can be used in combination with microposts to control cell shape or cell-to-cell interaction while measuring cellular forces. The protocols in this chapter describe how to make SU8 masters for stamps and microposts, how to use soft lithography to replicate these structures in polydimethylsiloxane, and how to functionalize the surface of the microposts for cell attachment. Copyright © 2014 Elsevier Inc. All rights reserved.

Detection of Drug-Resistant Mycobacterium tuberculosis.

PubMed

Engström, Anna; Juréen, Pontus

2015-01-01

Tuberculosis (TB) remains a global health problem. The increasing prevalence of drug-resistant Mycobacterium tuberculosis, the causative agent of TB, demands new measures to combat the situation. Rapid and accurate diagnosis of the pathogen and its drug susceptibility pattern is essential for timely initiation of optimal treatment, and, ultimately, control of the disease. We have developed a molecular method for detection of first- and second-line drug resistance in M. tuberculosis by Pyrosequencing(®). The method consists of seven Pyrosequencing assays for the detection of mutations in the genes or promoter regions, which are most commonly responsible for resistance to the drugs rifampicin, isoniazid, ethambutol, amikacin, kanamycin, capreomycin, and fluoroquinolones. The method was validated on clinical isolates and it was shown that the sensitivity and specificity of the method were comparable to those of Sanger sequencing. In the protocol in this chapter we describe the steps necessary for setting up and performing Pyrosequencing for M. tuberculosis. The first part of the protocol describes the assay development and the second part of the protocol describes utilization of the method.

Chapter 9: Metering Cross-Cutting Protocol. The Uniform Methods Project: Methods for Determining Energy-Efficiency Savings for Specific Measures

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

Mort, Dan

Estimated energy savings are calculated as the difference between the energy use during the baseline period and the energy use during the post installation period of the EEM. This chapter describes the physical properties measured in the process of evaluating EEMs and the specific metering methods for several types of measurements. Skill-level requirements and other operating considerations are discussed, including where, when, and how often measurements should be made. The subsequent section identifies metering equipment types and their respective measurement accuracies. This is followed by sections containing suggestions regarding proper data handling procedures and the categorization and definition of severalmore » load types.« less

Chapter 8: Whole-Building Retrofit with Consumption Data Analysis Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W.; Agnew, Ken; Goldberg, Mimi

Whole-building retrofits involve the installation of multiple measures. Whole-building retrofit programs take many forms. With a focus on overall building performance, these programs usually begin with an energy audit to identify cost-effective energy efficiency measures for the home. Measures are then installed, either at no cost to the homeowner or partially paid for by rebates and/or financing. The methods described here may also be applied to evaluation of single-measure retrofit programs. Related methods exist for replace-on-failure programs and for new construction, but are not the subject of this chapter.

Bioreactor Steroid Production and Analysis of Date Palm Embryogenic Callus.

PubMed

El-Sharabasy, Sherif; El-Dawayati, Maiada

2017-01-01

Several compounds and families of compounds of date palm secondary metabolites have been investigated. The analysis of date palm tissue has shown the abundance of secondary metabolites including phytosterols, e.g., steroids, an important group of pharmaceutical compounds. Biotechnology offers the opportunity to utilize cells, tissues, and organs grown in vitro and manipulated to obtain desired compounds. This chapter presents a protocol for the production, determination, and identification of steroids in date palm callus tissue. The addition of 0.01 mg/L pyruvic acid as a precursor to MS liquid culture medium enhances steroid production. In addition, the chapter describes the sterol analytical techniques based on gas-liquid chromatography and gas chromatography-mass spectrometry.

GMP-Compliant Expansion of Clinical-Grade Human Mesenchymal Stromal/Stem Cells Using a Closed Hollow Fiber Bioreactor.

PubMed

Barckhausen, Christina; Rice, Brent; Baila, Stefano; Sensebé, Luc; Schrezenmeier, Hubert; Nold, Philipp; Hackstein, Holger; Rojewski, Markus Thomas

2016-01-01

This chapter describes a method for GMP-compliant expansion of human mesenchymal stromal/stem cells (hMSC) from bone marrow aspirates, using the Quantum(®) Cell Expansion System from Terumo BCT. The Quantum system is a functionally closed, automated hollow fiber bioreactor system designed to reproducibly grow cells in either GMP or research laboratory environments. The chapter includes protocols for preparation of media, setup of the Quantum system, coating of the hollow fiber bioreactor, as well as loading, feeding, and harvesting of cells. We suggest a panel of quality controls for the starting material, the interim product, as well as the final product.

Modeling abundance using multinomial N-mixture models

USGS Publications Warehouse

Royle, Andy

2016-01-01

Multinomial N-mixture models are a generalization of the binomial N-mixture models described in Chapter 6 to allow for more complex and informative sampling protocols beyond simple counts. Many commonly used protocols such as multiple observer sampling, removal sampling, and capture-recapture produce a multivariate count frequency that has a multinomial distribution and for which multinomial N-mixture models can be developed. Such protocols typically result in more precise estimates than binomial mixture models because they provide direct information about parameters of the observation process. We demonstrate the analysis of these models in BUGS using several distinct formulations that afford great flexibility in the types of models that can be developed, and we demonstrate likelihood analysis using the unmarked package. Spatially stratified capture-recapture models are one class of models that fall into the multinomial N-mixture framework, and we discuss analysis of stratified versions of classical models such as model Mb, Mh and other classes of models that are only possible to describe within the multinomial N-mixture framework.

Chapter A6. Section 6.1. Temperature

USGS Publications Warehouse

Revised by Wilde, Franceska D.

2006-01-01

Accurate temperature measurements are required for accurate determinations of important environmental parameters such as pH, specific electrical conductance, and dissolved oxygen, and to the determination of chemical reaction rates and equilibria, biological activity, and physical fluid properties. This section of the National Field Manual (NFM) describes U.S. Geological Survey (USGS) guidance and protocols for measurement of temperature in air, ground water, and surface water and calibration of the equipment used.

Maize (Zea mays L.).

PubMed

Frame, Bronwyn; Warnberg, Katey; Main, Marcy; Wang, Kan

2015-01-01

Agrobacterium tumefaciens-mediated transformation is an effective method for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype Hi II. Our starting plant material is immature embryos cocultivated with an Agrobacterium strain carrying a standard binary vector. In addition to step-by-step laboratory transformation procedures, we include extensive details in growing donor plants and caring for transgenic plants in the greenhouse.

Agrobacterium- and Biolistic-Mediated Transformation of Maize B104 Inbred.

PubMed

Raji, Jennifer A; Frame, Bronwyn; Little, Daniel; Santoso, Tri Joko; Wang, Kan

2018-01-01

Genetic transformation of maize inbred genotypes remains non-routine for many laboratories due to variations in cell competency to induce embryogenic callus, as well as the cell's ability to receive and incorporate transgenes into the genome. This chapter describes two transformation protocols using Agrobacterium- and biolistic-mediated methods for gene delivery. Immature zygotic embryos of maize inbred B104, excised from ears harvested 10-14 days post pollination, are used as starting explant material. Disarmed Agrobacterium strains harboring standard binary vectors and the biolistic gun system Bio-Rad PDS-1000/He are used as gene delivery systems. The herbicide resistant bar gene and selection agent bialaphos are used for identifying putative transgenic type I callus events. Using the step-by-step protocols described here, average transformation frequencies (number of bialaphos resistant T 0 callus events per 100 explants infected or bombarded) of 4% and 8% can be achieved using the Agrobacterium- and biolistic-mediated methods, respectively. An estimated duration of 16-21 weeks is needed using either protocol from the start of transformation experiments to obtaining putative transgenic plantlets with established roots. In addition to laboratory in vitro procedures, detailed greenhouse protocols for producing immature ears as transformation starting material and caring for transgenic plants for seed production are also described.

Chapter 19: HVAC Controls (DDC/EMS/BAS) Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W.; Romberger, Jeff

The HVAC Controls Evaluation Protocol is designed to address evaluation issues for direct digital controls/energy management systems/building automation systems (DDC/EMS/BAS) that are installed to control heating, ventilation, and air-conditioning (HVAC) equipment in commercial and institutional buildings. (This chapter refers to the DDC/EMS/BAS measure as HVAC controls.) This protocol may also be applicable to industrial facilities such as clean rooms and labs, which have either significant HVAC equipment or spaces requiring special environmental conditions.

Micropropagation and organogenesis of Anthurium andreanum Lind cv Rubrun.

PubMed

Maira, Oropeza; Alexander, Mejías; Vargas, Teresa Edith

2010-01-01

Tissue culture techniques are routinely used for mass propagation and the establishment of disease free stock material. Virtually all pot type Anthuriums available in the market today are produced by tissue culture. In this chapter, we describe an efficient protocol to obtain Anthurium andreanum cv Rubrun vitro plants through micropropagation and organogenesis. Seeds from plant spadixes were germinated on MS medium supplemented with 0.5 mg/L BA. Micro-cuttings from in vitro germinated seedlings were subcultured on MS medium containing 2 mg/L BA and 0.5 mg/L NAA. Four-week-old in vitro plants obtained from microcuttings, showed callus proliferation at the stem base. The development of shoots and plantlets was observed from callus tissue. We also describe a detailed method for the histological analysis of callus tissue and a vitro plants acclimatization protocol.

Nanoparticle-mediated gene delivery.

PubMed

Jin, Sha; Leach, John C; Ye, Kaiming

2009-01-01

Nonviral gene delivery has been gaining considerable attention recently. Although the efficacy of DNA transfection, which is a major concern, is low in nonviral vector-mediated gene transfer compared with viral ones, nonviral vectors are relatively easy to prepare, less immunogenic and oncogenic, and have no potential of virus recombination and no limitation on the size of a transferred gene. The ability to incorporate genetic materials such as plasmid DNA, RNA, and siRNA into functionalized nanoparticles with little toxicity demonstrates a new era in pharmacotherapy for delivering genes selectively to tissues and cells. In this chapter, we highlight the basic concepts and applications of nonviral gene delivery using super paramagnetic iron oxide nanoparticles and functionalized silica nanoparticles. The experimental protocols related to these topics are described in the chapter.

Chapter A6. Section 6.4. pH

USGS Publications Warehouse

Wilde, Franceska D.; Busenberg, Eurybiades; Radtke, Dean B.

2006-01-01

Measurement of pH is critical to the understanding of the viability and vulnerability of environmental waters and is considered a master variable in determining the aqueous geochemistry of an aqueous system. pH is a measure that represents the hydrogen-ion concentration (activity) of a solution. This section of the National Field Manual (NFM) describes U.S. Geological Survey (USGS) guidance and protocols for measurement of pH in ground and surface waters.

The relationship of high school students' attitudes towards creation and evolution with the students' worldview philosophy

NASA Astrophysics Data System (ADS)

Ray, Donald David, Jr.

This prospectus examines the relationship of high school students' attitudes toward creation and evolution with the student's worldview philosophy. Chapter 1 presents the purpose of the research, which is to discover if there is a positive correlation in the relationship of high school students' attitudes toward creation and evolution and other aspects of worldview such as politics, economics, education, religion, and social issues. Also, the study will examine the frequency and comparative magnitude of the worldviews of homeschool, public school and Christian school students. Chapter 2 surveys precedents in the literature that are appropriate to the study. The significant literature to the study begins with the analysis of the relevant theological presuppositions pertaining to the topics of creation, evolution, and worldview. The analysis then moves to the literature on the relevant educational assumptions, which include theories, philosophies and practices related to high school students' attitudes toward creation and evolution with worldviews. Chapter 3 describes the methodological design for the study. Included in the chapter are the selection and profile of the sample, the selection and development of the instruments, namely the Creationist Worldview Test developed by Steve Deckard, and the PEERS Test---a worldview opinion survey developed by the Nehemiah Institute. Finally, the step-by-step protocol used in gathering the data completes the section. Chapter 4 presents the analysis of the data developed from the responses to the Creationist Worldview Test and the PEERS Test provided by the Nehemiah Institute and by the researcher. Profiles of each of the subgroups will be provided along with comparative analysis of the profiles. Concluding the chapter are summaries of profiles and comparative analysis. Chapter 5 presents a summary of the data describes the relationship of high school students' attitudes toward creation and evolution with the student's worldview philosophy. In addition, the major findings and general conclusions from the data are provided. The chapter concludes with recommendations for further research in this correlational analysis of origins.

Protocols for Biotechnological Interventions in Improvement of Vanilla (Vanilla planifolia Andrews.).

PubMed

Divakaran, Minoo; Babu, K Nirmal; Peter, K V

2016-01-01

Vanilla (Vanilla planifolia Andrews (syn. V. fragrans Salisb.), a native of Central America, is the primary source of natural vanillin and plays a major role in the global economy. The gene pool of vanilla is threatened by deforestation and overcollection that has resulted in disappearance of natural habitats and wild species. Continuous vegetative propagation and lack of natural seed set and sufficient variations in the gene pool hamper crop improvement programs. In vitro techniques, one of the key tools of plant biotechnology, can be employed for overcoming specific problems, viz. production of disease-free clones, inducing somaclonal variations, developing hybrids, gene pool conservation, incorporating desired traits by distant hybridization, genetic engineering, etc. However, realization of these objectives necessitates standardization of protocols. This chapter describes the various protocols optimized for crop improvement in Vanilla species.

Cultivar-Dependent Direct Organogenesis of Date Palm from Shoot Tip Explants.

PubMed

Abahmane, Larbi

2017-01-01

A number of public and private laboratories are working on date palm micropropagation to meet the increasing worldwide demand for date palm planting material. A standardized direct organogenesis protocol exists for the production of date palm plantlets to maintain the genetic fidelity of regenerated plants. Organogenesis has the advantage of using low concentrations of plant growth regulators and avoiding the callus phase. In addition, direct regeneration of vegetative buds minimizes the risk of somaclonal variation among plant regenerants. However, in vitro multiplication cycles should be limited in duration by frequent renewal of plant material. This chapter describes a simple and routine organogenesis protocol for date palm multiplication using shoot tip explants.

ChIP-seq and RNA-seq methods to study circadian control of transcription in mammals

PubMed Central

Takahashi, Joseph S.; Kumar, Vivek; Nakashe, Prachi; Koike, Nobuya; Huang, Hung-Chung; Green, Carla B.; Kim, Tae-Kyung

2015-01-01

Genome-wide analyses have revolutionized our ability to study the transcriptional regulation of circadian rhythms. The advent of next-generation sequencing methods has facilitated the use of two such technologies, ChIP-seq and RNA-seq. In this chapter, we describe detailed methods and protocols for these two techniques, with emphasis on their usage in circadian rhythm experiments in the mouse liver, a major target organ of the circadian clock system. Critical factors for these methods are highlighted and issues arising with time series samples for ChIP-seq and RNA-seq are discussed. Finally detailed protocols for library preparation suitable for Illumina sequencing platforms are presented. PMID:25662462

Detection of human Dicer and Argonaute 2 catalytic activity

PubMed Central

Perron, Marjorie P.; Landry, Patricia; Plante, Isabelle; Provost, Patrick

2013-01-01

The microRNA (miRNA)-guided RNA silencing pathway is a central and well-defined cellular process involved in messenger RNA (mRNA) translational control. This complex regulatory process is achieved by a well orchestrated machinery composed of a relatively few protein components, among which the ribonuclease III (RNase III) Dicer and Argonaute 2 (Ago2) play a central role. These two proteins are essential and it is of particular interest to measure and detect their catalytic activity under various situations and/or conditions. In this chapter, we describe different protocols that aim to study and determine the catalytic activity of Dicer and Ago2 in cell extracts, immune complexes and size-fractionated cell extracts. Another protocol aimed at assessing miRNA binding to Ago2 is also described. These experimental approaches are likely to be useful to researchers investigating the main steps of miRNA biogenesis and function in human health and diseases. PMID:21528451

An Action-Based Fine-Grained Access Control Mechanism for Structured Documents and Its Application

PubMed Central

Su, Mang; Li, Fenghua; Tang, Zhi; Yu, Yinyan; Zhou, Bo

2014-01-01

This paper presents an action-based fine-grained access control mechanism for structured documents. Firstly, we define a describing model for structured documents and analyze the application scenarios. The describing model could support the permission management on chapters, pages, sections, words, and pictures of structured documents. Secondly, based on the action-based access control (ABAC) model, we propose a fine-grained control protocol for structured documents by introducing temporal state and environmental state. The protocol covering different stages from document creation, to permission specification and usage control are given by using the Z-notation. Finally, we give the implementation of our mechanism and make the comparisons between the existing methods and our mechanism. The result shows that our mechanism could provide the better solution of fine-grained access control for structured documents in complicated networks. Moreover, it is more flexible and practical. PMID:25136651

An action-based fine-grained access control mechanism for structured documents and its application.

PubMed

Su, Mang; Li, Fenghua; Tang, Zhi; Yu, Yinyan; Zhou, Bo

2014-01-01

This paper presents an action-based fine-grained access control mechanism for structured documents. Firstly, we define a describing model for structured documents and analyze the application scenarios. The describing model could support the permission management on chapters, pages, sections, words, and pictures of structured documents. Secondly, based on the action-based access control (ABAC) model, we propose a fine-grained control protocol for structured documents by introducing temporal state and environmental state. The protocol covering different stages from document creation, to permission specification and usage control are given by using the Z-notation. Finally, we give the implementation of our mechanism and make the comparisons between the existing methods and our mechanism. The result shows that our mechanism could provide the better solution of fine-grained access control for structured documents in complicated networks. Moreover, it is more flexible and practical.

Engineering Poly(ethylene glycol) Materials to Promote Cardiogenesis

NASA Astrophysics Data System (ADS)

Smith, Amanda Walker

Heart failure is one of the leading causes of death worldwide, and the current costs of treatment put a significant economic burden on our societies. After an infarction, fibrotic tissue begins to form as part of the heart failure cascade. Current options to slow this process include a wide range of pharmaceutical agents, and ultimately the patient may require a heart transplant. Innovative treatment approaches are needed to bring down costs and improve quality of life. The possibility of regenerating or replacing damaged tissue with healthy cardiomyocytes is generating considerable excitement, but there are still many obstacles to overcome. First, while cell injections into the myocardium have demonstrated slight improvements in cardiac function, the actual engraftment of transplanted cells is very low. It is anticipated that improving engraftment will boost outcomes. Second, cellular differentiation and reprogramming protocols have not yet produced cells that are identical to adult cardiomyocytes, and immunogenicity continues to be a problem despite the advent of autologously derived induced pluripotent stem cells. This dissertation will explore biomaterials approaches to addressing these two obstacles. Tissue engineering scaffolds may improve cell engraftment by providing bioactive factors, preventing cell anoikis, and reducing cell washout by blood flow. Poly(ethylene glycol) (PEG) is often used as a coating to reduce implant rejection because it is highly resistant to protein adsorption. Because fibrosis of a material in contact with the myocardium could cause arrhythmias, PEG materials are highly relevant for cardiac tissue engineering applications. In Chapter 2, we describe a novel method for crosslinking PEG microspheres around cells to form a scaffold for tissue engineering. We then demonstrate that HL-1 cardiomyocyte viability and phenotype are retained throughout the fabrication process and during the first 7 weeks of culture. In the third chapter of the dissertation, we demonstrate that the use of PEG cell culture substrates can improve efficiency of direct reprogramming from fibroblasts to cardiomyocytes for cell transplantation. Standard tissue culture plastic adsorbs proteins from the cell media, increasing experimental variability via non-specific signaling. Because of its protein resistant properties, PEG provides cells with highly specific signals. In addition to improving the efficiency, we found that presentation of RGD peptides stimulated proliferation during reprogramming. Combined, the improvements enabled us to approximately double the number of cardiomyocytes produced by the protocol. In Chapter 4, we explore the effects of 3D culture on the direct reprogramming protocol described in Chapter 3. We demonstrate that the variables involved in 3D culture, including scaffold material, diffusion, cellular remodeling, and scaffold topography, have significant effects on reprogramming efficiency. This chapter provides the groundwork for future studies developing 3D microenvironments for efficient and scalable reprogramming to cardiomyocytes.

Computational Design of DNA-Binding Proteins.

PubMed

Thyme, Summer; Song, Yifan

2016-01-01

Predicting the outcome of engineered and naturally occurring sequence perturbations to protein-DNA interfaces requires accurate computational modeling technologies. It has been well established that computational design to accommodate small numbers of DNA target site substitutions is possible. This chapter details the basic method of design used in the Rosetta macromolecular modeling program that has been successfully used to modulate the specificity of DNA-binding proteins. More recently, combining computational design and directed evolution has become a common approach for increasing the success rate of protein engineering projects. The power of such high-throughput screening depends on computational methods producing multiple potential solutions. Therefore, this chapter describes several protocols for increasing the diversity of designed output. Lastly, we describe an approach for building comparative models of protein-DNA complexes in order to utilize information from homologous sequences. These models can be used to explore how nature modulates specificity of protein-DNA interfaces and potentially can even be used as starting templates for further engineering.

Live-cell imaging of mammalian RNAs with Spinach2.

PubMed

Strack, Rita L; Jaffrey, Samie R

2015-01-01

The ability to monitor RNAs of interest in living cells is crucial to understanding the function, dynamics, and regulation of this important class of molecules. In recent years, numerous strategies have been developed with the goal of imaging individual RNAs of interest in living cells, each with their own advantages and limitations. This chapter provides an overview of current methods of live-cell RNA imaging, including a detailed discussion of genetically encoded strategies for labeling RNAs in mammalian cells. This chapter then focuses on the development and use of "RNA mimics of GFP" or Spinach technology for tagging mammalian RNAs and includes a detailed protocol for imaging 5S and CGG60 RNA with the recently described Spinach2 tag. © 2015 Elsevier Inc. All rights reserved.

Application of SEM and EDX in studying biomineralization in plant tissues.

PubMed

He, Honghua; Kirilak, Yaowanuj

2014-01-01

This chapter describes protocols using formalin-acetic acid-alcohol (FAA) to fix plant tissues for studying biomineralization by means of scanning electron microscopy (SEM) and qualitative energy-dispersive X-ray microanalysis (EDX). Specimen preparation protocols for SEM and EDX mainly include fixation, dehydration, critical point drying (CPD), mounting, and coating. Gold-coated specimens are used for SEM imaging, while gold- and carbon-coated specimens are prepared for qualitative X-ray microanalyses separately to obtain complementary information on the elemental compositions of biominerals. During the specimen preparation procedure for SEM, some biominerals may be dislodged or scattered, making it difficult to determine their accurate locations, and light microscopy is used to complement SEM studies. Specimen preparation protocols for light microscopy generally include fixation, dehydration, infiltration and embedding with resin, microtome sectioning, and staining. In addition, microwave processing methods are adopted here to speed up the specimen preparation process for both SEM and light microscopy.

The Isolation of Pure Populations of Neurons by Laser Capture Microdissection: Methods and Application in Neuroscience.

PubMed

Morris, Renée; Mehta, Prachi

2018-01-01

In mammals, the central nervous system (CNS) is constituted of various cellular elements, posing a challenge to isolating specific cell types to investigate their expression profile. As a result, tissue homogenization is not amenable to analyses of motor neurons profiling as these represent less than 10% of the total spinal cord cell population. One way to tackle the problem of tissue heterogeneity and obtain meaningful genomic, proteomic, and transcriptomic profiling is to use laser capture microdissection technology (LCM). In this chapter, we describe protocols for the capture of isolated populations of motor neurons from spinal cord tissue sections and for downstream transcriptomic analysis of motor neurons with RT-PCR. We have also included a protocol for the immunological confirmation that the captured neurons are indeed motor neurons. Although focused on spinal cord motor neurons, these protocols can be easily optimized for the isolation of any CNS neurons.

Model Based Optimal Control, Estimation, and Validation of Lithium-Ion Batteries

NASA Astrophysics Data System (ADS)

Perez, Hector Eduardo

This dissertation focuses on developing and experimentally validating model based control techniques to enhance the operation of lithium ion batteries, safely. An overview of the contributions to address the challenges that arise are provided below. Chapter 1: This chapter provides an introduction to battery fundamentals, models, and control and estimation techniques. Additionally, it provides motivation for the contributions of this dissertation. Chapter 2: This chapter examines reference governor (RG) methods for satisfying state constraints in Li-ion batteries. Mathematically, these constraints are formulated from a first principles electrochemical model. Consequently, the constraints explicitly model specific degradation mechanisms, such as lithium plating, lithium depletion, and overheating. This contrasts with the present paradigm of limiting measured voltage, current, and/or temperature. The critical challenges, however, are that (i) the electrochemical states evolve according to a system of nonlinear partial differential equations, and (ii) the states are not physically measurable. Assuming available state and parameter estimates, this chapter develops RGs for electrochemical battery models. The results demonstrate how electrochemical model state information can be utilized to ensure safe operation, while simultaneously enhancing energy capacity, power, and charge speeds in Li-ion batteries. Chapter 3: Complex multi-partial differential equation (PDE) electrochemical battery models are characterized by parameters that are often difficult to measure or identify. This parametric uncertainty influences the state estimates of electrochemical model-based observers for applications such as state-of-charge (SOC) estimation. This chapter develops two sensitivity-based interval observers that map bounded parameter uncertainty to state estimation intervals, within the context of electrochemical PDE models and SOC estimation. Theoretically, this chapter extends the notion of interval observers to PDE models using a sensitivity-based approach. Practically, this chapter quantifies the sensitivity of battery state estimates to parameter variations, enabling robust battery management schemes. The effectiveness of the proposed sensitivity-based interval observers is verified via a numerical study for the range of uncertain parameters. Chapter 4: This chapter seeks to derive insight on battery charging control using electrochemistry models. Directly using full order complex multi-partial differential equation (PDE) electrochemical battery models is difficult and sometimes impossible to implement. This chapter develops an approach for obtaining optimal charge control schemes, while ensuring safety through constraint satisfaction. An optimal charge control problem is mathematically formulated via a coupled reduced order electrochemical-thermal model which conserves key electrochemical and thermal state information. The Legendre-Gauss-Radau (LGR) pseudo-spectral method with adaptive multi-mesh-interval collocation is employed to solve the resulting nonlinear multi-state optimal control problem. Minimum time charge protocols are analyzed in detail subject to solid and electrolyte phase concentration constraints, as well as temperature constraints. The optimization scheme is examined using different input current bounds, and an insight on battery design for fast charging is provided. Experimental results are provided to compare the tradeoffs between an electrochemical-thermal model based optimal charge protocol and a traditional charge protocol. Chapter 5: Fast and safe charging protocols are crucial for enhancing the practicality of batteries, especially for mobile applications such as smartphones and electric vehicles. This chapter proposes an innovative approach to devising optimally health-conscious fast-safe charge protocols. A multi-objective optimal control problem is mathematically formulated via a coupled electro-thermal-aging battery model, where electrical and aging sub-models depend upon the core temperature captured by a two-state thermal sub-model. The Legendre-Gauss-Radau (LGR) pseudo-spectral method with adaptive multi-mesh-interval collocation is employed to solve the resulting highly nonlinear six-state optimal control problem. Charge time and health degradation are therefore optimally traded off, subject to both electrical and thermal constraints. Minimum-time, minimum-aging, and balanced charge scenarios are examined in detail. Sensitivities to the upper voltage bound, ambient temperature, and cooling convection resistance are investigated as well. Experimental results are provided to compare the tradeoffs between a balanced and traditional charge protocol. Chapter 6: This chapter provides concluding remarks on the findings of this dissertation and a discussion of future work.

Blueberry (Vaccinium corymbosum L.).

PubMed

Song, Guo-Qing; Sink, Kenneth C

2006-01-01

Recent advances in plant biotechnology have led to a reliable and reproductive method for genetic transformation of blueberry. These efforts built on previous attempts at transient and stable transformation of blueberry that demonstrated the potential of Agrobacterium tumefaciens-mediated transformation, and as well, the difficulties of selecting and regenerating transgenic plants. As a prerequisite for successful stable transformation, efficient regeneration systems were required despite many reports on factors controlling shoot regeneration from leaf explants. The A. tumefaciens-mediated transformation protocol described in this chapter is based on combining efficient regeneration methods and the results of A. tumefaciens-mediated transient transformation studies to optimize selected parameters for gene transfer. The protocol has led to successful regeneration of transgenic plants of four commercially important highbush blueberry cultivars.

Cloning Endangered Felids by Interspecies Somatic Cell Nuclear Transfer.

PubMed

Gómez, Martha C; Pope, C Earle

2015-01-01

In 2003, the first wild felid was produced by interspecies somatic cell nuclear transfer. Since then other wild felid clone offspring have been produced by using the same technique with minor modifications. This chapter describes detailed protocols used in our laboratory for (1) the isolation, culture, and preparation of fibroblast cells as donor nucleus, and (2) embryo reconstruction with domestic cat enucleated oocytes to produce cloned embryos that develop to the blastocyst stage in vitro and, after transfer into synchronized recipients, establish successful pregnancies.

Formation of Stomach Tissue by Organoid Culture Using Mouse Embryonic Stem Cells.

PubMed

Noguchi, Taka-Aki K; Kurisaki, Akira

2017-01-01

In this chapter, we describe a method for the induction of stomach organoids from mouse embryonic stem (ES) cells. We used an embryoid body-based differentiation method to induce gastric primordial epithelium covered with mesenchyme and further differentiate it in Matrigel by 3D culture. The differentiated organoid contains both corpus- and antrum-specific mature gastric tissue cells. This protocol may be useful for a variety of studies in developmental biology and disease modeling of the stomach.

Telemetry Standards, IRIG Standard 106-17, Chapter 22, Network Based Protocol Suite

DTIC Science & Technology

2017-07-01

ER Fiber media connections using 10 Gigabit Ethernet shall comply with IEEE 802.3-2012, Section 5, Clause 52. 22.2.1.1.4 Auto- Negotiation ...22.2.1.1.4.1 Copper Auto- Negotiation Copper media connections, as described in the preceding sections, shall support auto- negotiation of speed, duplex, and...flow control in the manner specified in IEEE 802.3-2012, Section 2, Clause 28. 22.2.1.1.4.2 Fiber Auto- Negotiation Gigabit and 10 Gigabit fiber

Detection of Trypanosoma cruzi by Polymerase Chain Reaction.

PubMed

Márquez, María Elizabeth; Concepción, Juan Luis; González-Marcano, Eglys; Mondolfi, Alberto Paniz

2016-01-01

American Trypanosomiasis (Chagas disease) is an infectious disease caused by the hemoflagellate parasite Trypanosoma cruzi which is transmitted by reduviid bugs. T. cruzi infection occurs in a broad spectrum of reservoir animals throughout North, Central, and South America and usually evolves into an asymptomatic chronic clinical stage of the disease in which diagnosis is often challenging. This chapter describes the application of polymerase chain reaction (PCR) for the detection of Trypanosoma cruzi DNA including protocols for sample preparation, DNA extraction, and target amplification methods.

In vitro propagation of carnation (Dianthus caryophyllus L.).

PubMed

Casas, Jose L; Olmos, Enrique; Piqueras, Abel

2010-01-01

Carnation (Dianthus caryophyllus L.) is one of the most popular ornamental plants worldwide and also among the most studied ones, mainly in cut flower postharvest physiology. Several protocols for the in vitro propagation of this species including nodal segment culture, somatic embryogenesis, and adventitious shoot induction are described in this chapter. The presence of hyperhydricity as an abnormality during micropropagation of carnation plants has also been the object of research for many years and different strategies to overcome this problem are also included in this study.

Chapter 4: Small Commercial and Residential Unitary and Split System HVAC Heating and Cooling Equipment-Efficiency Upgrade Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Jacobson, David; Metoyer, Jarred

The specific measure described here involves improving the overall efficiency in air-conditioning systems as a whole (compressor, evaporator, condenser, and supply fan). The efficiency rating is expressed as the energy efficiency ratio (EER), seasonal energy efficiency ratio (SEER), and integrated energy efficiency ratio (IEER). The higher the EER, SEER or IEER, the more efficient the unit is.

Using Virtual Observatory Services in Sky View

NASA Technical Reports Server (NTRS)

McGlynn, Thomas A.

2007-01-01

For over a decade Skyview has provided astronomers and the public with easy access to survey and imaging data from all wavelength regimes. SkyView has pioneered many of the concepts that underlie the Virtual Observatory. Recently SkyView has been released as a distributable package which uses VO protocols to access image and catalog services. This chapter describes how to use the Skyview as a local service and how to customize it to access additional VO services and local data.

Chapter A7. Section 7.0. Five-Day Biochemical Oxygen Demand

USGS Publications Warehouse

Delzer, Gregory C.; McKenzie, Stuart W.

1999-01-01

The presence of a sufficient concentration of dissolved oxygen is critical to maintaining the aquatic life and aesthetic quality of streams and lakes. Determinng how organic matter affects the concentration of dissolved oxygen (DO) in a stream or lake is integral to water-quality management. The decay of organic matter in water is measured as biochemical or chemical oxygen demand. This report describes the field protocols used by U.S. Geological Survey (USGS) personnel to determine the five-day test for biochemical oxygen demand.

Using Environmental DNA for Invasive Species Surveillance and Monitoring.

PubMed

Mahon, Andrew R; Jerde, Christopher L

2016-01-01

The method employed for environmental DNA (eDNA) surveillance for detection and monitoring of rare species in aquatic systems has evolved dramatically since its first large-scale applications. Both active (targeted) and passive (total diversity) surveillance methods provide helpful information for management groups, but each has a suite of techniques that necessitate proper equipment training and use. The protocols described in this chapter represent some of the latest iterations in eDNA surveillance being applied in aquatic and marine systems.

Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.

PubMed

Asai, Akira; Takakuma, Kazuyuki

2017-01-01

When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.

Assessing the detection, reporting and investigation of adverse events in clinical trial protocols implemented in Cameroon: a documentary review of clinical trial protocols.

PubMed

Ebile, Akoh Walter; Ateudjieu, Jerome; Yakum, Martin Ndinakie; Djuidje, Marceline Ngounoue; Watcho, Pierre

2015-09-29

International guidelines recommend ethical and scientific quality standards for managing and reporting adverse events occurring during clinical trials to competent research ethics committees and regulatory authorities. The purpose of this study was to determine whether clinical trial protocols in Cameroon are developed in line with national requirements and international guidelines as far as detecting, reporting and investigating of adverse events is concerned. It was a documentary review of all approved clinical trial protocols that were submitted at the Cameroon National Ethics Committee for evaluation from 1997 through 2012. Data were extracted using a preconceived and validated grid. Protocol review process targeted the title, abstract, objectives, methodology, resources, and the chapter on safety. In total, 106 (4.9 %) clinical trial protocols were identified from 2173 protocols seen in the archive and 104 (4.8 %) included for review. Seventy six (73.1 %) trials did not include the surveillance of adverse events as part of their objective. A total of 91 (87.5 %) protocols did not budget for adverse event surveillance, 76 (73.1 %) did not have a data safety management board (DSMB), 11(10.6 %) included insurance for participants, 47 (45.2 %) did not include a case definition for serious adverse events, 33 (31.7 %) described procedures to detect adverse events, 33 (31.7 %) described procedure for reporting and 22 (21.2 %) described procedure for investigating adverse events. Most clinical trial protocols in Cameroon are developed to focus on benefits and pay little attention to harms. The development of national guidelines can improve the surveillance of adverse events in clinical trial research conducted in Cameroon. Adverse events surveillance tools and a budget are critical for an adequate planning for adverse event surveillance when developing trial protocols. Clinical trial protocols submitted in the Cameroon National Ethics Committee do not adequately plan to assess adverse events in clinical trial protocols. In order to improve on the safety of participants and marketed drug, there is a need to develop national guidelines for clinical trials by the government, and to improve evaluation procedures and monitoring of ongoing trials by the ethics committee.

Chapter 9. Benefits of International Collaboration | Science ...

EPA Pesticide Factsheets

In this chapter, we share what we have learned from working with our Brazilian colleagues on a multi university, multiyear, and multi basin ecological assessment and how those experiences were transmitted more broadly. These lessons (each of which is described in subsequent paragraphs) included 1) learning about markedly different ecosystems; 2) values to the U.S. Environmental Protection Agency (USEPA) of testing monitoring protocols in those ecosystems; 3) applying lessons from the CEMIG (Companhia Energética de Minas Gerais) project to research on other continents and elsewhere in Brazil; 4) advantages of academic team research; 5) benefits of corporate-sponsored research and federal student scholarships; 6) communicating with the general public; 7) the research web that has developed out of our work in Brazil; and 8) experiencing Brazilian culture. The USEPA’s NARS survey designs and field methods are being applied in large basin stream surveys in countries outside of the U.S. These applications not only provide valuable tests of the NARS approaches, but enhance International cooperation and generate new understandings of natural and anthropogenic controls on biota and physical habitat in streams. These understandings not only aid interpretation of the condition of streams in the regions surveyed, but also refine approaches for interpreting aquatic resource surveys elsewhere. In this book chapter, Robert Hughes and Philip Kaufmann describe th

Micropropagation systems of Feijoa (Acca sellowiana (O. Berg) Burret).

PubMed

Guerra, Miguel Pedro; Cangahuala-Inocente, Gabriela Claudia; Vesco, Lirio Luiz Dal; Pescador, Rosete; Caprestano, Clarissa Alves

2013-01-01

Acca sellowiana (O. Berg) Burret sin. Feijoa sellowiana (Myrtaceae) is a semiwoody fruit species native to South Brazil, Uruguay, and Argentina; edible fruits are tasty. The naturally occurring populations in Santa Catarina State show high variability in fruit size, color, and other features. A breeding program launched in 1990 resulted in the release of four Brazilian commercial varieties. The conventional clonal propagation methods of this species, such as cutting and grafting, have shown low efficiency. Therefore, tissue culture techniques were developed for mass propagation. This chapter describes several protocols based on organogenesis and somatic embryogenesis. Additional techniques including synthetic seed technology and temporary immersion system are also described.

Solid-State NMR Studies of Amyloid Materials: A Protocol to Define an Atomic Model of Aβ(1-42) in Amyloid Fibrils.

PubMed

Xiao, Yiling; McElheny, Dan; Hoshi, Minako; Ishii, Yoshitaka

2018-01-01

Intense efforts have been made to understand the molecular structures of misfolded amyloid β (Aβ) in order to gain insight into the pathological mechanism of Alzheimer's disease. Solid-state NMR spectroscopy (SSNMR) is considered a primary tool for elucidating the structures of insoluble and noncrystalline amyloid fibrils and other amyloid assemblies. In this chapter, we describe a detailed protocol to obtain the first atomic model of the 42-residue human Aβ peptide Aβ(1-42) in structurally homogeneous amyloid fibrils from our recent SSNMR study (Nat Struct Mol Biol 22:499-505, 2015). Despite great biological and clinical interest in Aβ(1-42) fibrils, their structural details have been long-elusive until this study. The protocol is divided into four sections. First, the solid-phase peptide synthesis (SPPS) and purification of monomeric Aβ(1-42) is described. We illustrate a controlled incubation method to prompt misfolding of Aβ(1-42) into homogeneous amyloid fibrils in an aqueous solution with fragmented Aβ(1-42) fibrils as seeds. Next, we detail analysis of Aβ(1-42) fibrils by SSNMR to obtain structural restraints. Finally, we describe methods to construct atomic models of Aβ(1-42) fibrils based on SSNMR results through two-stage molecular dynamics calculations.

Recommended protocols for sampling macrofungi

Treesearch

Gregory M. Mueller; John Paul Schmit; Sabine M. Hubndorf Leif Ryvarden; Thomas E. O' Dell; D. Jean Lodge; Patrick R. Leacock; Milagro Mata; Loengrin Umania; Qiuxin (Florence) Wu; Daniel L. Czederpiltz

2004-01-01

This chapter discusses several issues regarding reommended protocols for sampling macrofungi: Opportunistic sampling of macrofungi, sampling conspicuous macrofungi using fixed-size, sampling small Ascomycetes using microplots, and sampling a fixed number of downed logs.

Human Cells as Platform to Produce Gamma-Carboxylated Proteins.

PubMed

de Sousa Bomfim, Aline; de Freitas, Marcela Cristina Corrêa; Covas, Dimas Tadeu; de Sousa Russo, Elisa Maria

2018-01-01

The gamma-carboxylated proteins belong to a family of proteins that depend on vitamin K for normal biosynthesis. The major representative gamma-carboxylated proteins are the coagulation system proteins, for example, factor VII, factor IX, factor X, prothrombin, and proteins C, S, and Z. These molecules have harbored posttranslational modifications, such as glycosylation and gamma-carboxylation, and for this reason they need to be produced in mammalian cell lines. Human cells lines have emerged as the most promising alternative to the production of gamma-carboxylated proteins. In this chapter, the methods to generate human cells as a platform to produce gamma-carboxylated proteins, for example the coagulation factors VII and IX, are presented. From the cell line modification up to the vitamin K adaptation of the produced cells is described in the protocols presented in this chapter.

Refractive index measurements of single, spherical cells using digital holographic microscopy.

PubMed

Schürmann, Mirjam; Scholze, Jana; Müller, Paul; Chan, Chii J; Ekpenyong, Andrew E; Chalut, Kevin J; Guck, Jochen

2015-01-01

In this chapter, we introduce digital holographic microscopy (DHM) as a marker-free method to determine the refractive index of single, spherical cells in suspension. The refractive index is a conclusive measure in a biological context. Cell conditions, such as differentiation or infection, are known to yield significant changes in the refractive index. Furthermore, the refractive index of biological tissue determines the way it interacts with light. Besides the biological relevance of this interaction in the retina, a lot of methods used in biology, including microscopy, rely on light-tissue or light-cell interactions. Hence, determining the refractive index of cells using DHM is valuable in many biological applications. This chapter covers the main topics that are important for the implementation of DHM: setup, sample preparation, and analysis. First, the optical setup is described in detail including notes and suggestions for the implementation. Following that, a protocol for the sample and measurement preparation is explained. In the analysis section, an algorithm for the determination of quantitative phase maps is described. Subsequently, all intermediate steps for the calculation of the refractive index of suspended cells are presented, exploiting their spherical shape. In the last section, a discussion of possible extensions to the setup, further measurement configurations, and additional analysis methods are given. Throughout this chapter, we describe a simple, robust, and thus easily reproducible implementation of DHM. The different possibilities for extensions show the diverse fields of application for this technique. Copyright © 2015 Elsevier Inc. All rights reserved.

Lipids and Fatty Acids in Algae: Extraction, Fractionation into Lipid Classes, and Analysis by Gas Chromatography Coupled with Flame Ionization Detector (GC-FID).

PubMed

Guihéneuf, Freddy; Schmid, Matthias; Stengel, Dagmar B

2015-01-01

Despite the number of biochemical studies exploring algal lipids and fatty acid biosynthesis pathways and profiles, analytical methods used by phycologists for this purpose are often diverse and incompletely described. Potential confusion and potential variability of the results between studies can therefore occur due to change of protocols for lipid extraction and fractionation, as well as fatty acid methyl esters (FAME) preparation before gas chromatography (GC) analyses. Here, we describe a step-by-step procedure for the profiling of neutral and polar lipids using techniques such as solid-liquid extraction (SLE), thin-layer chromatography (TLC), and gas chromatography coupled with flame ionization detector (GC-FID). As an example, in this protocol chapter, analyses of neutral and polar lipids from the marine microalga Pavlova lutheri (an EPA/DHA-rich haptophyte) will be outlined to describe the distribution of fatty acid residues within its major lipid classes. This method has been proven to be a reliable technique to assess changes in lipid and fatty acid profiles in several other microalgal species and seaweeds.

Optimal molecular profiling of tissue and tissue components: defining the best processing and microdissection methods for biomedical applications.

PubMed

Rodriguez-Canales, Jaime; Hanson, Jeffrey C; Hipp, Jason D; Balis, Ulysses J; Tangrea, Michael A; Emmert-Buck, Michael R; Bova, G Steven

2013-01-01

Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of any tissue-based biological phenomenon. This updated chapter reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genotype. The quality of samples isolated from tissue and used for molecular analysis is often glossed over or omitted from publications, making interpretation and replication of data difficult or impossible. Fortunately, recently developed techniques allow life scientists to better document and control the quality of samples used for a given assay, creating a foundation for improvement in this area. Tissue processing for molecular studies usually involves some or all of the following steps: tissue collection, gross dissection/identification, fixation, processing/embedding, storage/archiving, sectioning, staining, microdissection/annotation, and pure analyte labeling/identification and quantification. We provide a detailed comparison of some current tissue microdissection technologies and provide detailed example protocols for tissue component handling upstream and downstream from microdissection. We also discuss some of the physical and chemical issues related to optimal tissue processing and include methods specific to cytology specimens. We encourage each laboratory to use these as a starting point for optimization of their overall process of moving from collected tissue to high-quality, appropriately anatomically tagged scientific results. Improvement in this area will significantly increase life science quality and productivity. The chapter is divided into introduction, materials, protocols, and notes subheadings. Because many protocols are covered in each of these sections, information relating to a single protocol is not contiguous. To get the greatest benefit from this chapter, readers are advised to read through the entire chapter first, identify protocols appropriate to their laboratory for each step in their workflow, and then reread entries in each section pertaining to each of these single protocols.

Sun and Sky Radiance Measurements and Data Analysis Protocols. Chapter 5

NASA Technical Reports Server (NTRS)

Frouin, Robert; Holben, Brent; Miller, Mark; Pietras, Christophe; Porter, John; Voss, Ken

2001-01-01

This chapter is concerned with two types of radiometric measurements essential to verify atmospheric correction algorithms and to calibrate vicariously satellite ocean color sensors. The first type is a photometric measurement of the direct solar beam to determine the optical thickness of the atmosphere. The intensity of the solar beam can be measured directly, or obtained indirectly from measurements of diffuse global upper hemispheric irradiance. The second type is a measurement of the solar aureole and sky radiance distribution using a CCD camera, or a scanning radiometer viewing in and perpendicular to the solar principal plane. From the two types of measurements, the optical properties of aerosols, highly variable in space and time, can be derived. Because of the high variability, the aerosol properties should be known at the time of satellite overpass. Atmospheric optics measurements, however, are not easy to perform at sea, from a ship or any platform. This complicates the measurement protocols and data analysis. Some instrumentation cannot be deployed at sea, and is limited to island and coastal sites. In the following, measurement protocols are described for radiometers commonly used to measure direct atmospheric transmittance and sky radiance, namely standard sun photometers, fast-rotating shadow-band radiometers, automated sky scanning systems, and CCD cameras. Methods and procedures to analyze and quality control the data are discussed, as well as proper measurement strategies for evaluation of atmospheric correction algorithms and satellite-derived ocean color.

Microcalli Induction in Protoplasts Isolated from Embryogenic Callus of Date Palm.

PubMed

Titouh, Khayreddine; Boufis, Nazim; Khelifi, Lakhdar

2017-01-01

Date palm (Phoenix dactylifera L.) production is severely hampered due to several pests and diseases. Biotechnological tools such as protoplast fusion appear as an alternative to ensure rapid genetic improvement and multiplication of this species. However, establishment of an effective system of plant regeneration from protoplasts culture is a prerequisite for date palm somatic hybridization. In this chapter, we describe an effective protocol to induce microcalli in protoplasts isolated from nodular callus of important Algerian date palm cultivars. In this protocol, the main factors influencing the isolation (i.e., enzymatic solution, mannitol concentration, duration, and mode of maceration) of protoplasts from the calli of Algerian date palm cultivars were optimized. Purified protoplasts were cultured on a semisolid medium supplemented with a hormonal balance of auxin and cytokinin to obtain microcalli formation.

Plasmid fermentation process for DNA immunization applications.

PubMed

Carnes, Aaron E; Williams, James A

2014-01-01

Plasmid DNA for immunization applications must be of the highest purity and quality. The ability of downstream purification to efficiently produce a pure final product is directly influenced by the performance of the upstream fermentation process. While several clinical manufacturing facilities already have validated fermentation processes in place to manufacture plasmid DNA for use in humans, a simple and inexpensive laboratory-scale fermentation process can be valuable for in-house production of plasmid DNA for use in animal efficacy studies. This chapter describes a simple fed-batch fermentation process for producing bacterial cell paste enriched with high-quality plasmid DNA. A constant feeding strategy results in a medium cell density culture with continuously increasing plasmid amplification towards the end of the process. Cell banking and seed culture preparation protocols, which can dramatically influence final product yield and quality, are also described. These protocols are suitable for production of research-grade plasmid DNA at the 100 mg-to-1.5 g scale from a typical 10 L laboratory benchtop fermentor.

Orchids (Cymbidium spp., Oncidium, and Phalaenopsis).

PubMed

Chan, Ming-Tsair; Chan, Yuan-Li; Sanjaya

2006-01-01

Recent advances in genetic engineering have made the transformation and regeneration of plants into a powerful tool for orchid improvement. This chapter presents a simple and reproducible Agrobacterium tumefaciens-mediated transformation protocol and molecular screening technique of transgenics for two orchid species, Oncidium and Phalaenopsis. The target tissues for gene transfer were protocorm-like bodies (PLBs) derived from protocorms, into which constructed foreign genes were successfully introduced. To establish stable transformants, two stages of selection were applied on the PLBs co-cultivated with A. tumefaciens. About 10% transformation efficiency was achieved in Oncidium orchid, as 108 antibiotic resistant independent PLBs were proliferated from 1000 infected PLBs. In Phalaenopsis orchid about 11 to 12% of transformation efficiency was achieved by using the present protocol. Different molecular methods and GUS-staining used to screen putative transgenic plants to confirm the integration of foreign DNA into the orchid genome were also described in detail. The methods described would also be useful for transformation of desired genes into other orchid species.

Cyclotide isolation and characterization.

PubMed

Craik, David J; Henriques, Sonia Troeira; Mylne, Joshua S; Wang, Conan K

2012-01-01

Cyclotides are disulfide-rich cyclic peptides produced by plants with the presumed natural function of defense agents against insect pests. They are present in a wide range of plant tissues, being ribosomally synthesized via precursor proteins that are posttranslationally processed to produce mature peptides with a characteristic cyclic backbone and cystine knot motif associated with their six conserved cysteine residues. Their processing is not fully understood but involves asparaginyl endoproteinase activity. In addition to interest in their defense roles and their unique topologies, cyclotides have attracted attention as potential templates in peptide-based drug design applications. This chapter provides protocols for the isolation of cyclotides from plants, their detection and sequencing by mass spectrometry, and their structural analysis by NMR, as well as describing methods for the isolation of nucleic acid sequences that encode their precursor proteins. Assays to assess their membrane-binding interactions are also described. These protocols provide a "starter kit" for researchers entering the cyclotide field. Copyright © 2012 Elsevier Inc. All rights reserved.

Imaging of Lipids and Metabolites Using Nanospray Desorption Electrospray Ionization Mass Spectrometry

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

Lanekoff, Ingela; Laskin, Julia

In recent years, mass spectroscopy imaging (MSI) has emerged as a foundational technique in metabolomics and drug screening providing deeper understanding of complex mechanistic pathways within biochemical systems and biological organisms. We have been invited to contribute a chapter to a new Springer series volume, entitled “Mass Spectrometry Imaging of Small Molecules”. The volume is planned for the highly successful lab protocol series Methods in Molecular Biology, published by Humana Press, USA. The volume is aimed to equip readers with step-by-step mass spectrometric imaging protocols and bring rapidly maturing methods of MS imaging to life science researchers. The chapter willmore » provide a detailed protocol of ambient MSI by use of nanospray desorption electrospray ionization.« less

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Revised

NASA Technical Reports Server (NTRS)

Fargion, Giulietta S.; Mueller, James L.

2000-01-01

The document stipulates protocols for measuring bio-optical and radiometric data for the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project activities and algorithm development. This document supersedes the earlier version (Mueller and Austin 1995) published as Volume 25 in the SeaWiFS Technical Report Series. This document marks a significant departure from, and improvement on, theformat and content of Mueller and Austin (1995). The authorship of the protocols has been greatly broadened to include experts specializing in some key areas. New chapters have been added to provide detailed and comprehensive protocols for stability monitoring of radiometers using portable sources, abovewater measurements of remote-sensing reflectance, spectral absorption measurements for discrete water samples, HPLC pigment analysis and fluorometric pigment analysis. Protocols were included in Mueller and Austin (1995) for each of these areas, but the new treatment makes significant advances in each topic area. There are also new chapters prescribing protocols for calibration of sun photometers and sky radiance sensors, sun photometer and sky radiance measurements and analysis, and data archival. These topic areas were barely mentioned in Mueller and Austin (1995).

Extraction and Estimation of Secondary Metabolites from Date Palm Cell Suspension Cultures.

PubMed

Naik, Poornananda M; Al-Khayri, Jameel M

2017-01-01

The health benefits of dates arise from their content of phytochemicals, known for having pharmacological properties, including flavonoids, carotenoids, phenolic acids, sterols, procyanidins, and anthocyanins. In vitro cell culture technology has become an attractive means for the production of biomass and bioactive compounds. This chapter describes step-by-step procedures for the induction and proliferation of callus from date palm offshoots on Murashige and Skoog (MS) medium supplemented with plant growth regulators. Subsequently cell suspension cultures are established for optimum biomass accumulation, based on the growth curve developed by packed cell volume as well as fresh and dry weights. The highest production of biomass occurs at the 11th week after culturing. Moreover, this chapter describes methodologies for the extraction and analysis of secondary metabolites of date palm cell suspension cultures using high-performance liquid chromatography (HPLC). The optimum level of catechin, caffeic acid, apigenin, and kaempferol from the cell suspension cultures establishes after the 11th and 12th weeks of culture. This protocol is useful for scale-up production of secondary metabolites from date palm cell suspension cultures.

Protein 3-Nitrotyrosine in Complex Biological Samples: Quantification by High-Pressure Liquid Chromatography/Electrochemical Detection and Emergence of Proteomic Approaches for Unbiased Identification of Modification Sites

PubMed Central

Nuriel, Tal; Deeb, Ruba S.; Hajjar, David P.; Gross, Steven S.

2008-01-01

Nitration of tyrosine residues by nitric oxide (NO)-derived species results in the accumulation of 3-nitrotyrosine in proteins, a hallmark of nitrosative stress in cells and tissues. Tyrosine nitration is recognized as one of the multiple signaling modalities used by NO-derived species for the regulation of protein structure and function in health and disease. Various methods have been described for the quantification of protein 3-nitrotyrosine residues, and several strategies have been presented toward the goal of proteome-wide identification of protein tyrosine modification sites. This chapter details a useful protocol for the quantification of 3-nitrotyrosine in cells and tissues using high-pressure liquid chromatography with electrochemical detection. Additionally, this chapter describes a novel biotin-tagging strategy for specific enrichment of 3-nitrotyrosine-containing peptides. Application of this strategy, in conjunction with high-throughput MS/MS-based peptide sequencing, is anticipated to fuel efforts in developing comprehensive inventories of nitrosative stress-induced protein-tyrosine modification sites in cells and tissues. PMID:18554526

Method for somatic cell nuclear transfer in zebrafish.

PubMed

Siripattarapravat, Kannika; Cibelli, Jose B

2011-01-01

Somatic cell nuclear transfer (SCNT) has been a well-known technique for decades and widely applied to generate identical animals, including ones with genetic alterations. The system has been demonstrated successfully in zebrafish. The elaborated requirements of SCNT, however, limit reproducibility of the established model to a few groups in zebrafish research community. In this chapter, we meticulously outline each step of the published protocol as well as preparations of equipments and reagents used in zebrafish SCNT. All describable detailed-tips are elaborated in texts and figures. Copyright © 2011 Elsevier Inc. All rights reserved.

Chikungunya Virus Infection of Aedes Mosquitoes.

PubMed

Wong, Hui Vern; Chan, Yoke Fun; Sam, I-Ching; Sulaiman, Wan Yusof Wan; Vythilingam, Indra

2016-01-01

In vivo infection of mosquitoes is an important method to study and characterize arthropod-borne viruses. Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that is transmitted primarily by Aedes mosquitoes. In this chapter, we describe a protocol for infection of CHIKV in two species of Aedes mosquitoes, Aedes aegypti and Aedes albopictus, together with the isolation of CHIKV in different parts of the infected mosquito such as midgut, legs, wings, salivary gland, head, and saliva. This allows the study of viral infection, replication and dissemination within the mosquito vector.

Fabrication and Handling of 3D Scaffolds Based on Polymers and Decellularized Tissues.

PubMed

Shpichka, Anastasia; Koroleva, Anastasia; Kuznetsova, Daria; Dmitriev, Ruslan I; Timashev, Peter

2017-01-01

Polymeric, ceramic and hybrid material-based three-dimensional (3D) scaffold or matrix structures are important for successful tissue engineering. While the number of approaches utilizing the use of cell-based scaffold and matrix structures is constantly growing, it is essential to provide a framework of their typical preparation and evaluation for tissue engineering. This chapter describes the fabrication of 3D scaffolds using two-photon polymerization, decellularization and cell encapsulation methods and easy-to-use protocols allowing assessing the cell morphology, cytotoxicity and viability in these scaffolds.

Micropropagation of Lavandula spp.

PubMed

Gonçalves, Sandra; Romano, Anabela

2013-01-01

Lavandula species are some of the most popular ornamental and medicinal plants with great economic values. These species are vegetative propagated by stem cuttings. However, the poor rooting ability and vulnerability of plantlets to contamination are major limiting factors for propagation. In vitro culture methods are suitable to overcome these limitations. This chapter describes protocols for in vitro propagation of Lavandula viridis L'Hér and Lavandula vera DC. Nodal shoot proliferation of L. viridis and plant regeneration from leaf-derived callus of L. vera by an "open culture system" are highlighted.

Design and Fabrication of N-Alkyl-Polyethylenimine-Stabilized Iron Oxide Nanoclusters for Gene Delivery

PubMed Central

Liu, Gang; Wang, Zhiyong; Lee, Seulki; Ai, Hua; Chen, Xiaoyuan

2013-01-01

With the rapid development of nanotechnology, inorganic magnetic nanoparticles, especially iron oxide nanoparticles (IOs), have emerged as great vehicles for biomedical diagnostic and therapeutic applications. In order to rationally design IO-based gene delivery nanovectors, surface modification is essential and determines the loading and release of the gene of interest. Here we highlight the basic concepts and applications of nonviral gene delivery vehicles based on low molecular weight N-alkyl polyethylenimine-stabilized IOs. The experimental protocols related to these topics are described in this chapter. PMID:22568910

Generation of Stable Knockout Mammalian Cells by TALEN-Mediated Locus-Specific Gene Editing.

PubMed

Mahata, Barun; Biswas, Kaushik

2017-01-01

Precise and targeted genome editing using Transcription Activator-Like Effector Endonucleases (TALENs) has been widely used and proven to be an extremely effective and specific knockout strategy in both cultured cells and animal models. The current chapter describes a protocol for the construction and generation of TALENs using serial and hierarchical digestion and ligation steps, and using the synthesized TALEN pairs to achieve locus-specific targeted gene editing in mammalian cell lines using a modified clonal selection strategy in an easy and cost-efficient manner.

Analysis of the regulation of viral transcription.

PubMed

Gloss, Bernd; Kalantari, Mina; Bernard, Hans-Ulrich

2005-01-01

Despite the small genomes and number of genes of papillomaviruses, regulation of their transcription is very complex and governed by numerous transcription factors, cis-responsive elements, and epigenetic phenomena. This chapter describes the strategies of how one can approach a systematic analysis of these factors, elements, and mechanisms. From the numerous different techniques useful for studying transcription, we describe in detail three selected protocols of approaches that have been relevant in shaping our knowledge of human papillomavirus transcription. These are DNAse I protection ("footprinting") for location of transcription-factor binding sites, electrophoretic mobility shifts ("gelshifts") for analysis of bound transcription factors, and bisulfite sequencing for analysis of DNA methylation as a prerequisite for epigenetic transcriptional regulation.

Visualization of DNA and Protein-DNA Complexes with Atomic Force Microscopy

PubMed Central

Lyubchenko, Yuri L.; Gall, Alexander A.; Shlyakhtenko, Luda S.

2014-01-01

This article describes sample preparation techniques for AFM imaging of DNA and protein–DNA complexes. The approach is based on chemical functionalization of the mica surface with aminopropyl silatrane (APS) to yield an APS-mica surface. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations, and in a broad range of pH. The chapter describes the methodologies for the preparation of APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and purifi cation of APS is also provided. The AFM applications are illustrated with examples of images of DNA and protein–DNA complexes. PMID:24357372

Isolation and Assessment of Stability of Six Formulations of Entomopathogenic Beauveria bassiana.

PubMed

Mwamburi, Lizzy A

2016-01-01

Beauveria bassiana is the most widely studied and exploited entomopathogen. The development of a suitable formulation for B. bassiana is a critical component in aiding the entomopathogen germinate and infect the host. In addition to being economical to produce, having high residual activity, it is also important that the formulation is easy to handle, stable during storage, and convenient to mix and apply and be consistently effective in controlling the target pest.In this chapter we describe preparation of experimental formulations of conidia of B. bassiana. The formulations are prepared with barley, rice, wheat bran, clay, kaolin, and peat. The protocol for assessing the stability of the formulations of B. bassiana is also described.

Use of Humanized RS-ATL8 Reporter System for Detection of Allergen-Specific IgE Sensitization in Human Food Allergy.

PubMed

Ali, Eman Ali; Nakamura, Ryosuke; Falcone, Franco H

2017-01-01

Allergen-specific Immunoglobulin E (IgE) determination lies at the heart of diagnosis of sensitization to food and other allergens. In the past few years, reporter systems capable of detecting the presence of allergen-specific IgE have been developed by several labs. These rely on humanized rat basophil leukemia cell lines stably transfected with reporter genes such as firefly luciferase. In this chapter, we describe protocols for the use of the RS-ATL8 cell line (IgE cross-linking-induced luciferase expression; EXiLE) in 96-well and 384-well formats. We also describe optional treatment steps for enveloped virus and complement inactivation.

Real-time imaging of specific genomic loci in eukaryotic cells using the ANCHOR DNA labelling system.

PubMed

Germier, Thomas; Sylvain, Audibert; Silvia, Kocanova; David, Lane; Kerstin, Bystricky

2018-06-01

Spatio-temporal organization of the cell nucleus adapts to and regulates genomic processes. Microscopy approaches that enable direct monitoring of specific chromatin sites in single cells and in real time are needed to better understand the dynamics involved. In this chapter, we describe the principle and development of ANCHOR, a novel tool for DNA labelling in eukaryotic cells. Protocols for use of ANCHOR to visualize a single genomic locus in eukaryotic cells are presented. We describe an approach for live cell imaging of a DNA locus during the entire cell cycle in human breast cancer cells. Copyright © 2018 Elsevier Inc. All rights reserved.

Micropropagation of African violet (Saintpaulia ionantha Wendl.).

PubMed

Shukla, Mukund; Sullivan, J Alan; Jain, Shri Mohan; Murch, Susan J; Saxena, Praveen K

2013-01-01

Micropropagation is an important tool for rapid multiplication and the creation of genetic variability in African violets (Saintpaulia ionantha Wendl.). Successful in vitro propagation depends on the specific requirements and precise manipulation of various factors such as the type of explants used, physiological state of the mother plant, plant growth regulators in the culture medium, and growth conditions. Development of cost-effective protocols with a high rate of multiplication is a crucial requirement for commercial application of micropropagation. The current chapter describes an optimized protocol for micropropagation of African violets using leaf explants obtained from in vitro grown plants. In this process, plant regeneration occurs via both somatic embryogenesis and shoot organogenesis simultaneously in the explants induced with the growth regulator thidiazuron (TDZ; N-phenyl-N'-1,2,3-thidiazol-5-ylurea). The protocol is simple, rapid, and efficient for large-scale propagation of African violet and the dual routes of regeneration allow for multiple applications of the technology from simple clonal propagation to induction or selection of variants to the production of synthetic seeds.

In Vitro Mass Propagation of Cymbopogon citratus Stapf., a Medicinal Gramineae.

PubMed

Quiala, Elisa; Barbón, Raúl; Capote, Alina; Pérez, Naivy; Jiménez, Elio

2016-01-01

Cymbopogon citratus (D.C.) Stapf. is a medicinal plant source of lemon grass oils with multiple uses in the pharmaceutical and food industry. Conventional propagation in semisolid culture medium has become a fast tool for mass propagation of lemon grass, but the production cost must be lower. A solution could be the application of in vitro propagation methods based on liquid culture advantages and automation. This chapter provides two efficient protocols for in vitro propagation via organogenesis and somatic embryogenesis of this medicinal plant. Firstly, we report the production of shoots using a temporary immersion system (TIS). Secondly, a protocol for somatic embryogenesis using semisolid culture for callus formation and multiplication, and liquid culture in a rotatory shaker and conventional bioreactors for the maintenance of embryogenic culture, is described. Well-developed plants can be achieved from both protocols. Here we provide a fast and efficient technology for mass propagation of this medicinal plant taking the advantage of liquid culture and automation.

Chapter 6: Residential Lighting Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Dimetrosky, Scott; Parkinson, Katie

Given new regulations, increased complexity in the market, and the general shift from CFLs to LEDs, this evaluation protocol was updated in 2017 to shift the focus of the protocols toward LEDs and away from CFLs and to resolve evaluation uncertainties affecting residential lighting incentive programs.

USDA-ARS GRACEnet Project Protocols, Chapter 3. Chamber-based trace gas flux measurements4

USDA-ARS?s Scientific Manuscript database

This protocol addresses N2O, CO2 and CH4 flux measurement by soil chamber methodology. The reactivities of other gasses of interest such as NOx O3, CO, and NH3 will require different chambers and associated instrumentation. Carbon dioxide is included as an analyte with this protocol; however, when p...

Chapter 15: Preparations of entomopathogens and diseased specimens for more detailed study using microscopy

USDA-ARS?s Scientific Manuscript database

The science of Insect Pathology encompasses a diverse assemblage of pathogens from a large and varied group of hosts. Microscopy techniques and protocols for these organisms are complex and varied and often require modifications and adaptations of standard procedures. The objective of this chapter...

Methods to study the biogenesis of membrane proteins in yeast mitochondria.

PubMed

Weckbecker, Daniel; Herrmann, Johannes M

2013-01-01

The biogenesis of mitochondrial membrane proteins is an intricate process that relies on the import and submitochondrial sorting of nuclear-encoded preproteins and on the synthesis of mitochondrial translation products in the matrix. Subsequently, these polypeptides need to be inserted into the outer and the inner membranes of the organelle where many of them assemble into multisubunit complexes. In this chapter we provide established protocols to study these different processes experimentally using mitochondria of budding yeast. In particular, methods are described in detail to purify mitochondria, to study mitochondrial protein synthesis, to follow the import of radiolabeled preproteins into isolated mitochondria, and to assess membrane association and the aggregation of mitochondrial proteins by fractionation. These protocols and a list of dos and don'ts shall enable beginners and experienced scientists to address the targeting and assembly of mitochondrial membrane proteins.

An introduction to planar chromatography and its application to natural products isolation.

PubMed

Gibbons, Simon

2012-01-01

Thin-layer chromatography (TLC) is an easy, inexpensive, rapid, and the most widely used method for the analysis and isolation of small organic natural and synthetic products. It also has use in the biological evaluation of organic compounds, particularly in the areas of antimicrobial and antioxidant metabolites and for the evaluation of acetylcholinesterase inhibitors which have utility in the treatment of Alzheimer's disease. The ease and inexpensiveness of use of this technique, coupled with the ability to rapidly develop separation and bioassay protocols will ensure that TLC will be used for some considerable time alongside conventional instrumental methods. This chapter deals with the basic principles of TLC and describes methods for the analysis and isolation of natural products. Examples of methods for isolation of several classes of natural product are detailed and protocols for TLC bioassays are given.

High-pressure freezing for scanning transmission electron tomography analysis of cellular organelles.

PubMed

Walther, Paul; Schmid, Eberhard; Höhn, Katharina

2013-01-01

Using an electron microscope's scanning transmission mode (STEM) for collection of tomographic datasets is advantageous compared to bright field transmission electron microscopic (TEM). For image formation, inelastic scattering does not cause chromatic aberration, since in STEM mode no image forming lenses are used after the beam has passed the sample, in contrast to regular TEM. Therefore, thicker samples can be imaged. It has been experimentally demonstrated that STEM is superior to TEM and energy filtered TEM for tomography of samples as thick as 1 μm. Even when using the best electron microscope, adequate sample preparation is the key for interpretable results. We adapted protocols for high-pressure freezing of cultivated cells from a physiological state. In this chapter, we describe optimized high-pressure freezing and freeze substitution protocols for STEM tomography in order to obtain high membrane contrast.

Ion-Exchange Chromatography: Basic Principles and Application.

PubMed

Cummins, Philip M; Rochfort, Keith D; O'Connor, Brendan F

2017-01-01

Ion-Exchange Chromatography (IEC) allows for the separation of ionizable molecules on the basis of differences in charge properties. Its large sample-handling capacity, broad applicability (particularly to proteins and enzymes), moderate cost, powerful resolving ability, and ease of scale-up and automation have led to it becoming one of the most versatile and widely used of all liquid chromatography (LC) techniques. In this chapter, we review the basic principles of IEC, as well as the broader criteria for selecting IEC conditions. By way of further illustration, we outline basic laboratory protocols to partially purify a soluble serine peptidase from bovine whole brain tissue, covering crude tissue extract preparation through to partial purification of the target enzyme using anion-exchange chromatography. Protocols for assaying total protein and enzyme activity in both pre- and post-IEC fractions are also described.

Biolistic- and Agrobacterium-mediated transformation protocols for wheat.

PubMed

Tamás-Nyitrai, Cecília; Jones, Huw D; Tamás, László

2012-01-01

After rice, wheat is considered to be the most important world food crop, and the demand for high-quality wheat flour is increasing. Although there are no GM varieties currently grown, wheat is an important target for biotechnology, and we anticipate that GM wheat will be commercially available in 10-15 years. In this chapter, we summarize the main features and challenges of wheat transformation and then describe detailed protocols for the production of transgenic wheat plants both by biolistic and Agrobacterium-mediated DNA-delivery. Although these methods are used mainly for bread wheat (Triticum aestivum L.), they can also be successfully applied, with slight modifications, to tetraploid durum wheat (T. turgidum L. var. durum). The appropriate size and developmental stage of explants (immature embryo-derived scutella), the conditions to produce embryogenic callus tissues, and the methods to regenerate transgenic plants under increasing selection pressure are provided in the protocol. To illustrate the application of herbicide selection system, we have chosen to describe the use of the plasmid pAHC25 for biolistic transformation, while for Agrobacterium-mediated transformation the binary vector pAL156 (incorporating both the bar gene and the uidA gene) has been chosen. Beside the step-by-step methodology for obtaining stably transformed and normal fertile plants, procedures for screening and testing transgenic wheat plants are also discussed.

Chapter 20: Data Center IT Efficiency Measures Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Huang, Robert; Masanet, Eric

This chapter focuses on IT measures in the data center and examines the techniques and analysis methods used to verify savings that result from improving the efficiency of two specific pieces of IT equipment: servers and data storage.

Vocational Training in Portugal.

ERIC Educational Resources Information Center

Mota, Artur; And Others

This report on vocational training in Portugal contains 8 chapters, a 26-item bibliography, and 2 appendices. Chapter 1 describes the geography, political system, population, and employment situation of Portugal. Chapter 2 describes the economic framework of Portugal. Chapter 3 describes the education system, the apprenticeship system, other…

Dissecting Transcriptional Heterogeneity in Pluripotency: Single Cell Analysis of Mouse Embryonic Stem Cells.

PubMed

Guedes, Ana M V; Henrique, Domingos; Abranches, Elsa

2016-01-01

Mouse Embryonic Stem cells (mESCs) show heterogeneous and dynamic expression of important pluripotency regulatory factors. Single-cell analysis has revealed the existence of cell-to-cell variability in the expression of individual genes in mESCs. Understanding how these heterogeneities are regulated and what their functional consequences are is crucial to obtain a more comprehensive view of the pluripotent state.In this chapter we describe how to analyze transcriptional heterogeneity by monitoring gene expression of Nanog, Oct4, and Sox2, using single-molecule RNA FISH in single mESCs grown in different cell culture medium. We describe in detail all the steps involved in the protocol, from RNA detection to image acquisition and processing, as well as exploratory data analysis.

Algorithm Updates for the Fourth SeaWiFS Data Reprocessing

NASA Technical Reports Server (NTRS)

Hooker, Stanford, B. (Editor); Firestone, Elaine R. (Editor); Patt, Frederick S.; Barnes, Robert A.; Eplee, Robert E., Jr.; Franz, Bryan A.; Robinson, Wayne D.; Feldman, Gene Carl; Bailey, Sean W.

2003-01-01

The efforts to improve the data quality for the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) data products have continued, following the third reprocessing of the global data set in May 2000. Analyses have been ongoing to address all aspects of the processing algorithms, particularly the calibration methodologies, atmospheric correction, and data flagging and masking. All proposed changes were subjected to rigorous testing, evaluation and validation. The results of these activities culminated in the fourth reprocessing, which was completed in July 2002. The algorithm changes, which were implemented for this reprocessing, are described in the chapters of this volume. Chapter 1 presents an overview of the activities leading up to the fourth reprocessing, and summarizes the effects of the changes. Chapter 2 describes the modifications to the on-orbit calibration, specifically the focal plane temperature correction and the temporal dependence. Chapter 3 describes the changes to the vicarious calibration, including the stray light correction to the Marine Optical Buoy (MOBY) data and improved data screening procedures. Chapter 4 describes improvements to the near-infrared (NIR) band correction algorithm. Chapter 5 describes changes to the atmospheric correction and the oceanic property retrieval algorithms, including out-of-band corrections, NIR noise reduction, and handling of unusual conditions. Chapter 6 describes various changes to the flags and masks, to increase the number of valid retrievals, improve the detection of the flag conditions, and add new flags. Chapter 7 describes modifications to the level-la and level-3 algorithms, to improve the navigation accuracy, correct certain types of spacecraft time anomalies, and correct a binning logic error. Chapter 8 describes the algorithm used to generate the SeaWiFS photosynthetically available radiation (PAR) product. Chapter 9 describes a coupled ocean-atmosphere model, which is used in one of the changes described in Chapter 4. Finally, Chapter 10 describes a comparison of results from the third and fourth reprocessings along the US. Northeast coast.

Alfalfa (Medicago sativa L.).

PubMed

Fu, Chunxiang; Hernandez, Timothy; Zhou, Chuanen; Wang, Zeng-Yu

2015-01-01

Alfalfa (Medicago sativa L.) is a high-quality forage crop widely grown throughout the world. This chapter describes an efficient protocol that allows for the generation of large number of transgenic alfalfa plants by sonication-assisted Agrobacterium-mediated transformation. Binary vectors carrying different selectable marker genes that confer resistance to phosphinothricin (bar), kanamycin (npt II), or hygromycin (hph) were used to generate transgenic alfalfa plants. Intact trifoliates collected from clonally propagated plants in the greenhouse were sterilized with bleach and then inoculated with Agrobacterium strain EHA105. More than 80 % of infected leaf pieces could produce rooted transgenic plants in 4-5 months after Agrobacterium-mediated transformation.

Generation and Functional Evaluation of Designer Monoterpene Synthases.

PubMed

Srividya, N; Lange, I; Lange, B M

2016-01-01

Monoterpene synthases are highly versatile enzymes that catalyze the first committed step in the pathways toward terpenoids, the structurally most diverse class of plant natural products. Recent advancements in our understanding of the reaction mechanism have enabled engineering approaches to develop mutant monoterpene synthases that produce specific monoterpenes. In this chapter, we are describing protocols to introduce targeted mutations, express mutant enzyme catalysts in heterologous hosts, and assess their catalytic properties. Mutant monoterpene synthases have the potential to contribute significantly to synthetic biology efforts aimed at producing larger amounts of commercially attractive monoterpenes. © 2016 Elsevier Inc. All rights reserved.

Chapter A6. Section 6.7. Turbidity

USGS Publications Warehouse

Anderson, Chauncey W.

2005-01-01

Turbidity is one of the indicators used to assess the environmental health of water bodies. Turbidity is caused by the presence of suspended and dissolved matter, such as clay, silt, finely divided organic matter, plankton and other microscopic organisms, organic acids, and dyes. This section of the National Field Manual (NFM) describes the USGS protocols for determining turbidity in surface and ground waters, including extensive guidance for equipment selection and data reporting. It includes the revised approach to turbidity measurement and reporting that was implemented by the U.S. Geological Survey (USGS) in October 2004 to account for technological advances and consequent measurement complexities.

Design and fabrication of N-alkyl-polyethylenimine-stabilized iron oxide nanoclusters for gene delivery.

PubMed

Liu, Gang; Wang, Zhiyong; Lee, Seulki; Ai, Hua; Chen, Xiaoyuan

2012-01-01

With the rapid development of nanotechnology, inorganic magnetic nanoparticles, especially iron oxide nanoparticles (IOs), have emerged as great vehicles for biomedical diagnostic and therapeutic applications. In order to rationally design IO-based gene delivery nanovectors, surface modification is essential and determines the loading and release of the gene of interest. Here we highlight the basic concepts and applications of nonviral gene delivery vehicles based on low molecular weight N-alkyl polyethylenimine-stabilized IOs. The experimental protocols related to these topics are described in this chapter. Copyright © 2012 Elsevier Inc. All rights reserved.

Genetic Transformation of Switchgrass

NASA Astrophysics Data System (ADS)

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Genetic transformation of switchgrass.

PubMed

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

2009-01-01

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Methods to Study the Role of Progranulin in the Tumor Microenvironment.

PubMed

Elkabets, Moshe; Brook, Samuel

2018-01-01

Accurate measurement of progranulin (PGRN) in the circulation and in the tumor microenvironment is essential for understanding its role in cancer progression and metastasis. This chapter describes a number of approaches to measure the transcription level of the GRN gene and to detect and analyze PGRN expression in cancer cells and in the local environment of the tumor, in mouse and human samples. These validated protocols are utilized to investigate the functional role of PGRN in cancer. Finally, we discuss strategies to investigate the functions of PGRN in tumors using genetically modified mouse models and gene silencing techniques.

Exploring the Cytoskeleton During Intracytoplasmic Sperm Injection in Humans

NASA Astrophysics Data System (ADS)

Rawe, Vanesa Y.; Chemes, Héctor

Understanding the cellular events during fertilization in mammals is a major challenge that can contribute to the improvement of future infertility treatments in humans and reproductive performance in farm animals. Of special interest is the role of the oocyte and sperm cytoskeleton during the initial interaction between gametes. The aim of this chapter is to describe methods for studying cytoskeletal features during in vitro fertilization after intracytoplasmic sperm injection (ICSI) in humans. The following protocols will provide a detailed description of how to perform immunodetection and imaging of human eggs, zygotes, and sperm by fluorescence (confocal and epifluorescence) and electron microscopy.

Evaluation of the Insecticidal Efficacy of Wild Type and Recombinant Baculoviruses.

PubMed

Popham, Holly J R; Ellersieck, Mark R; Li, Huarong; Bonning, Bryony C

2016-01-01

A considerable amount of work has been undertaken to genetically enhance the efficacy of baculovirus insecticides. Following construction of a genetically altered baculovirus, laboratory bioassays are used to quantify various parameters of insecticidal activity such as the median lethal concentration (or dose) required to kill 50 % of infected larvae (LC50 or LD50), median survival of larvae infected (ST50), and feeding damage incurred by infected larvae. In this chapter, protocols are described for a variety of bioassays and the corresponding data analyses for assessment of the insecticidal activity of baculovirus insecticides.

3D Culture for Self-Formation of the Cerebellum from Human Pluripotent Stem Cells Through Induction of the Isthmic Organizer.

PubMed

Muguruma, Keiko

2017-01-01

Pluripotent stem cells (PSCs) possess self-organizing abilities in 3D culture. This property has been demonstrated in recent studies, including the generation of various neuroectodermal and endodermal tissues. For example, PSCs are able to differentiate into specific type of neural tissues, such as the neocortex and the optic cup, in response to local positional information brought about by signals during embryogenesis. In contrast, the generation of cerebellar tissue from PSCs requires a secondary induction by a signaling center, called the isthmic organizer, which first appears in the cell aggregate in 3D culture. Such developmental complexity of cerebellum has hampered establishment of effective differentiation culture system from PSCs, thus far.We recently reported that cerebellar neurons are generated from human PSCs (hPSCs). In this chapter, we describe an efficient protocol for differentiation of 3D cerebellar neuroepithelium from hPSCs. We also describe the protocols for further differentiation into specific neurons in the cerebellar cortex, such as Purkinje cells and the granule cells.

Analysis of the conductivity of plasmodesmata by microinjection.

PubMed

Kragler, Friedrich

2015-01-01

Pressure microinjection can be used to introduce fluorescent dyes and labeled macromolecules into single cells. The method allows measuring transport activity of macromolecules such as proteins and RNA molecules within and between cells. Routinely, plant mesophyll cells are injected with fluorescent dextran molecules of specific sizes to measure an increase of the size exclusion limit of plasmodesmata in the presence of a co-injected or expressed protein. The mobility of a macromolecule can also be addressed directly by injecting a recombinant protein that itself is labeled with fluorescent dye and following its transport to neighboring cells. This chapter describes a pressure microinjection protocol successfully applied to Nicotiana leaves. This protocol requires basic skills and experience in handling a microscope equipped with an imaging system, a micromanipulator, and a microinjection system attached to an upright microscope. Using this equipment, a trained person can inject approximately 10-20 mesophyll cells per hour.

Protocol for the use of a rapid real-time PCR method for the detection of HIV-1 proviral DNA using double-stranded primer.

PubMed

Pau, Chou-Pong; Wells, Susan K; Granade, Timothy C

2012-01-01

This chapter describes a real-time PCR method for the detection of HIV-1 proviral DNA in whole blood samples using a novel double-stranded primer system. The assay utilizes a simple commercially available DNA extraction method and a rapid and easy-to-perform real-time PCR protocol to consistently detect a minimum of four copies of HIV-1 group M proviral DNA in as little as 90 min after sample (whole blood) collection. Co-amplification of the human RNase P gene serves as an internal control to monitor the efficiency of both the DNA extraction and amplification. Once the assay is validated properly, it may be suitable as an alternative confirmation test for HIV-1 infections in a variety of HIV testing venues including the mother-to-child transmission testing sites, clinics, and diagnostic testing centers.

Blueberry (Vaccinium corymbosum L.).

PubMed

Song, Guo-Qing

2015-01-01

Vaccinium consists of approximately 450 species, of which highbush blueberry (Vaccinium corymbosum) is one of the three major Vaccinium fruit crops (i.e., blueberry, cranberry, and lingonberry) domesticated in the twentieth century. In blueberry the adventitious shoot regeneration using leaf explants has been the most desirable regeneration system to date; Agrobacterium tumefaciens-mediated transformation is the major gene delivery method and effective selection has been reported using either the neomycin phosphotransferase II gene (nptII) or the bialaphos resistance (bar) gene as selectable markers. The A. tumefaciens-mediated transformation protocol described in this chapter is based on combining the optimal conditions for efficient plant regeneration, reliable gene delivery, and effective selection. The protocol has led to successful regeneration of transgenic plants from leaf explants of four commercially important highbush blueberry cultivars for multiple purposes, providing a powerful approach to supplement conventional breeding methods for blueberry by introducing genes of interest.

Protocol for the Production of a Vaccine Against Argentinian Hemorrhagic Fever.

PubMed

Ambrosio, Ana María; Mariani, Mauricio Andrés; Maiza, Andrea Soledad; Gamboa, Graciela Susana; Fossa, Sebastián Edgardo; Bottale, Alejando Javier

2018-01-01

Argentinian hemorrhagic Fever (AHF) is a febrile, acute disease caused by Junín virus (JUNV), a member of the Arenaviridae. Different approaches to obtain an effective antigen to prevent AHF using complete live or inactivated virus, as well as molecular constructs, have reached diverse development stages. This chapter refers to JUNV live attenuated vaccine strain Candid #1, currently used in Argentina to prevent AHF. A general standardized protocol used at Instituto Nacional de Enfermedades Virales Humanas (Pergamino, Pcia. Buenos Aires, Argentina) to manufacture the tissue culture derived Candid #1 vaccine is described. Intermediate stages like viral seeds and cell culture bank management, bulk vaccine manufacture, and finished product processing are also separately presented in terms of Production and Quality Control/Quality Assurance requirements, under the Adminitracion Nacional de Medicamentos, Alimentos y Tecnología Medica (ANMAT), the Argentine national regulatory authority.

Encapsulation of Date Palm Somatic Embryos: Synthetic Seeds.

PubMed

Bekheet, Shawky A

2017-01-01

Synthetic seed or encapsulated somatic embryos may be used for propagation, storage, and exchange of plant germplasm and have many diverse applications in date palm cultivation. They have advantages over conventional use of offshoot material for germplasm propagation, maintenance, exchange, and transportation. This chapter describes a protocol for date palm synthetic seed production by encapsulation of somatic embryos with sodium alginate. Among three concentrations used, 3% sodium alginate followed by dropping into 2.5% calcium chloride (CaCl 2 ) solution shows the best concentration of gel matrix for both maintenance and recovery. In addition, storage of the encapsulated date palm somatic embryos at 5 °C improves the survival and conversion into plantlets; otherwise, 20 g/L sucrose in the culture medium enhances conversion of the recovered somatic embryos to plantlets. This protocol is promising for in vitro conservation and international exchange of date palm germplasm.

Etiquette and Protocol: A Guide for Campus Events.

ERIC Educational Resources Information Center

Harris, April L.

Intended for special events planners on college campuses, this book offers advice on matters of etiquette and protocol for campus events. Chapters cover the following topics: (1) invitations (e.g., the precedence of extending invitations, invitation components, formal invitations, types of invitations); (2) forms of address (with examples of…

Methods in Molecular Biology Mouse Genetics: Methods and Protocols | Center for Cancer Research

Cancer.gov

Mouse Genetics: Methods and Protocols provides selected mouse genetic techniques and their application in modeling varieties of human diseases. The chapters are mainly focused on the generation of different transgenic mice to accomplish the manipulation of genes of interest, tracing cell lineages, and modeling human diseases.

EPICS Input Output Controller (IOC) Record Reference Manual

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

Anderson, J.B.; Kraimer, M.R.

1994-12-01

This manual describes all supported EPICS record types. The first chapter gives introduction and describes the field summary table. The second chapter describes the fields in database common, i.e. the fields that are present in every record type. The third chapter describes the input and output field that are common to many record types and have the same usage wherever they are used. Following the third chapter is a separate chapter for each record type containing a description of all the fields for that record type except those in database common.

Characterization of rock samples and mineralogical controls on leachates

USGS Publications Warehouse

Hammarstrom, Jane M.; Cravotta, Charles A.; Galeone, Daniel G.; Jackson, John C.; Dulong, Frank T.; Hornberger, Roger J.; Brady, Keith B.C.

2009-01-01

Rocks associated with coal beds typically include shale, sandstone, and (or) limestone. In addition to common rock-forming minerals, all of these rock types may contain sulfide and sulfate minerals, various carbonate minerals, and organic material. These different minerals have inherently different solubility characteristics, as well as different acid-generating or acid-neutralizing potentials. The abundance and composition of sulfur- and carbonate-bearing minerals are of particular interest in interpreting the leaching column data because (1) pyrite and carbonate minerals are the primary controls on the acid-base account of a sample, (2) these minerals incorporate trace metals that can be released during weathering, and (3) these minerals readily react during weathering due to mineral dissolution and oxidation of iron.Rock samples were collected by the Pennsylvania Department of Environmental Protection (PaDEP) from five different sites to assess the draft standardized leaching column method (ADTI-WP2) for the prediction of weathering rates and water quality at coal mines. Samples were sent to USGS laboratories for mineralogical characterization and to ActLabs for chemical analysis. The samples represent a variety of rock types (shales, sandstones, and coal refuse) that are typical of coal overburden in the eastern United States. These particular samples were chosen for testing the weathering protocols because they represent a range of geochemical and lithologic characteristics, sulfur contents, and acid-base accounting characteristics (Hornberger et al., 2003). The rocks contain variable amounts of pyrite and carbonate minerals and vary in texture.This chapter includes bulk rock chemical data and detailed mineralogical and textural data for unweathered starting materials used in the interlaboratory validation study, and for two samples used in the early phases of leaching column tests (Wadesville Sandstone, Leechburg Coal Refuse). We also characterize some of the post-weathering rock samples, report trace-element content in leachate, and discuss mineralogical controls on leachate quality based on data from one of the participating laboratories. Table 5.1 lists the samples described in this chapter, the sample numbers, and comments on the characteristics of each lithology. Sample locations are plotted in Figure 5.1. Chapters 2 and 3 describe the sample locations, sample preparation protocols, ABA characteristics, and rationale for selection of rock samples for testing. Microprobe data for pyrite and carbonate minerals are tabulated in Appendix 5.1. Leachate data, along with a series of graphs showing concentration and cumulative transport trends, for the laboratory data discussed in this chapter are included as Excel spreadsheets in Appendices 5.2 and 5.3. Leach column data for the interlaboratory study are evaluated and interpreted in Chapters 7 -11.

A mass spectrometry primer for mass spectrometry imaging

PubMed Central

Rubakhin, Stanislav S.; Sweedler, Jonathan V.

2011-01-01

Mass spectrometry imaging (MSI), a rapidly growing subfield of chemical imaging, employs mass spectrometry (MS) technologies to create single- and multi-dimensional localization maps for a variety of atoms and molecules. Complimentary to other imaging approaches, MSI provides high chemical specificity and broad analyte coverage. This powerful analytical toolset is capable of measuring the distribution of many classes of inorganics, metabolites, proteins and pharmaceuticals in chemically and structurally complex biological specimens in vivo, in vitro, and in situ. The MSI approaches highlighted in this Methods in Molecular Biology volume provide flexibility of detection, characterization, and identification of multiple known and unknown analytes. The goal of this chapter is to introduce investigators who may be unfamiliar with MS to the basic principles of the mass spectrometric approaches as used in MSI. In addition to guidelines for choosing the most suitable MSI method for specific investigations, cross-references are provided to the chapters in this volume that describe the appropriate experimental protocols. PMID:20680583

Multiparameter Cell Cycle Analysis.

PubMed

Jacobberger, James W; Sramkoski, R Michael; Stefan, Tammy; Woost, Philip G

2018-01-01

Cell cycle cytometry and analysis are essential tools for studying cells of model organisms and natural populations (e.g., bone marrow). Methods have not changed much for many years. The simplest and most common protocol is DNA content analysis, which is extensively published and reviewed. The next most common protocol, 5-bromo-2-deoxyuridine S phase labeling detected by specific antibodies, is also well published and reviewed. More recently, S phase labeling using 5'-ethynyl-2'-deoxyuridine incorporation and a chemical reaction to label substituted DNA has been established as a basic, reliable protocol. Multiple antibody labeling to detect epitopes on cell cycle regulated proteins, which is what this chapter is about, is the most complex of these cytometric cell cycle assays, requiring knowledge of the chemistry of fixation, the biochemistry of antibody-antigen reactions, and spectral compensation. However, because this knowledge is relatively well presented methodologically in many papers and reviews, this chapter will present a minimal Methods section for one mammalian cell type and an extended Notes section, focusing on aspects that are problematic or not well described in the literature. Most of the presented work involves how to segment the data to produce a complete, progressive, and compartmentalized cell cycle analysis from early G1 to late mitosis (telophase). A more recent development, using fluorescent proteins fused with proteins or peptides that are degraded by ubiquitination during specific periods of the cell cycle, termed "Fucci" (fluorescent, ubiquitination-based cell cycle indicators) provide an analysis similar in concept to multiple antibody labeling, except in this case cells can be analyzed while living and transgenic organisms can be created to perform cell cycle analysis ex or in vivo (Sakaue-Sawano et al., Cell 132:487-498, 2007). This technology will not be discussed.

Overcoming Hurdles Implementing Multi-skilling Policies

DTIC Science & Technology

2015-03-26

skilled workforce? Chapter II will communicate important concepts found in the literature on skill proficiency topics. These topics include skill...training methods that might improve learning and retention during the acquisition phase. 10 The active interlock modeling (AIM) protocol is a dyadic ...retention, as found in 43 Chapter 2. These techniques include dyadic training methods, overlearning, feedback, peer support, and managerial support

Insect cells-baculovirus system for the production of difficult to express proteins.

PubMed

Osz-Papai, Judit; Radu, Laura; Abdulrahman, Wassim; Kolb-Cheynel, Isabelle; Troffer-Charlier, Nathalie; Birck, Catherine; Poterszman, Arnaud

2015-01-01

The production of sufficient quantities of homogenous protein not only is an essential prelude for structural investigations but also represents a rate-limiting step for many human functional studies. Although technologies for expression of recombinant proteins and complexes have been improved tremendously, in many cases, protein production remains a challenge and can be associated with considerable investment. This chapter describes simple and efficient protocols for expression screening and optimization of protein production in insect cells using the baculovirus expression system. We describe the procedure, starting from the cloning of a gene of interest into an expression transfer baculovirus vector, followed by generation of the recombinant virus by homologous recombination, evaluation of protein expression, and scale-up. Handling of insect cell cultures and preparation of bacmid for co-transfection are also detailed.

Experimental Induction of Genome Chaos.

PubMed

Ye, Christine J; Liu, Guo; Heng, Henry H

2018-01-01

Genome chaos, or karyotype chaos, represents a powerful survival strategy for somatic cells under high levels of stress/selection. Since the genome context, not the gene content, encodes the genomic blueprint of the cell, stress-induced rapid and massive reorganization of genome topology functions as a very important mechanism for genome (karyotype) evolution. In recent years, the phenomenon of genome chaos has been confirmed by various sequencing efforts, and many different terms have been coined to describe different subtypes of the chaotic genome including "chromothripsis," "chromoplexy," and "structural mutations." To advance this exciting field, we need an effective experimental system to induce and characterize the karyotype reorganization process. In this chapter, an experimental protocol to induce chaotic genomes is described, following a brief discussion of the mechanism and implication of genome chaos in cancer evolution.

Constructing Tissue Microarrays: Protocols and Methods Considering Potential Advantages and Disadvantages for Downstream Use.

PubMed

Bingle, Lynne; Fonseca, Felipe P; Farthing, Paula M

2017-01-01

Tissue microarrays were first constructed in the 1980s but were used by only a limited number of researchers for a considerable period of time. In the last 10 years there has been a dramatic increase in the number of publications describing the successful use of tissue microarrays in studies aimed at discovering and validating biomarkers. This, along with the increased availability of both manual and automated microarray builders on the market, has encouraged even greater use of this novel and powerful tool. This chapter describes the basic techniques required to build a tissue microarray using a manual method in order that the theory behind the practical steps can be fully explained. Guidance is given to ensure potential disadvantages of the technique are fully considered.

47 CFR 12.3 - 911 and E911 analyses and reports.

Code of Federal Regulations, 2013 CFR

2013-10-01

... wireless 911 rules set forth in § 20.18 of this chapter; and interconnected Voice over Internet Protocol... Public Safety and Homeland Security Bureau (PSHSB) has the delegated authority to implement and activate... generation Internet Protocol-based E911 platform. (b) These reports are due 120 days from the date that the...

21 CFR 660.36 - Samples and protocols.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Samples and protocols. 660.36 Section 660.36 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS... Research Sample Custodian (ATTN: HFM-672) (see mailing addresses in § 600.2 of this chapter), within 30...

Pull-down Assay to Characterize Ca2+/Calmodulin Binding to Plant Receptor Kinases.

PubMed

Kaufmann, Christine; Sauter, Margret

2017-01-01

Plant receptor-like kinases (RLKs) are regulated by posttranscriptional modification and by interaction with regulatory proteins. A common modification of RLKs is (auto)phosphorylation, and a common regulatory protein is the calcium sensor calmodulin (CaM). We have developed protocols to detect the interaction of an RLK with CaM. The interaction with CaM was shown by bimolecular fluorescence complementation (BiFC) (see Chapter 14) and pull-down assay (this chapter). Both methods offer unique advantages. BiFC is useful in showing interaction of soluble as well as of membrane-bound proteins in planta. Pull-down assays are restricted to soluble proteins and provide in vitro data. The pull-down assay provides the advantage that proteins can be modified prior to binding and that experimental conditions such as the concentration of Ca 2+ or other divalent cations can be controlled. This chapter provides a pull-down protocol to study RLK-CaM interaction with optional steps to investigate the impact of RLK phosphorylation or of Ca 2+ .

Public opinion about energy: a literature review

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

Farhar, B.C.; Weis, P.; Unseld, C.T.

1979-06-01

A review and analysis of 115 surveys of the general population between 1973 and 1978 (before the Three Mile Island accident, sudden gasoline price increases, and spot gasoline shortages) are presented. The survey data are organized and analyzed according to a set of categories described in Chapter 2. The public's definition of the energy situation is the focus of Chapter 3. Chapter 4 describes public opinion concerning energy conservation pertaining to transportation and domestic energy use -- whether it is efficacious, how knowledgeable people are about conservation, whether they favor it, and whether they engage in it. Findings on themore » solar energy alternative are described in Chapter 5. Public opinion concerning more conventional sources of energy -- oil and natural gas, coal, and nuclear energy -- are discussed in Chapter 6. In Chapter 7, a summary of analytical findings by 10 major sociodemographic characteristics is presented. Chapter 8 describes analytical findings using various social-psychological variables. A summary is presented in Chapter 9.« less

Live-Cell Imaging of Mitochondria and the Actin Cytoskeleton in Budding Yeast.

PubMed

Higuchi-Sanabria, Ryo; Swayne, Theresa C; Boldogh, Istvan R; Pon, Liza A

2016-01-01

Maintenance and regulation of proper mitochondrial dynamics and functions are necessary for cellular homeostasis. Numerous diseases, including neurodegeneration and muscle myopathies, and overall cellular aging are marked by declining mitochondrial function and subsequent loss of multiple other cellular functions. For these reasons, optimized protocols are needed for visualization and quantification of mitochondria and their function and fitness. In budding yeast, mitochondria are intimately associated with the actin cytoskeleton and utilize actin for their movement and inheritance. This chapter describes optimal approaches for labeling mitochondria and the actin cytoskeleton in living budding yeast cells, for imaging the labeled cells, and for analyzing the resulting images.

Magnetic Characterization of Iron Oxide Nanoparticles for Biomedical Applications.

PubMed

Maldonado-Camargo, Lorena; Unni, Mythreyi; Rinaldi, Carlos

2017-01-01

Iron oxide nanoparticles are of interest in a wide range of biomedical applications due to their response to applied magnetic fields and their unique magnetic properties. Magnetization measurements in constant and time-varying magnetic field are often carried out to quantify key properties of iron oxide nanoparticles. This chapter describes the importance of thorough magnetic characterization of iron oxide nanoparticles intended for use in biomedical applications. A basic introduction to relevant magnetic properties of iron oxide nanoparticles is given, followed by protocols and conditions used for measurement of magnetic properties, along with examples of data obtained from each measurement, and methods of data analysis.

The production of multiprotein complexes in insect cells using the baculovirus expression system.

PubMed

Abdulrahman, Wassim; Radu, Laura; Garzoni, Frederic; Kolesnikova, Olga; Gupta, Kapil; Osz-Papai, Judit; Berger, Imre; Poterszman, Arnaud

2015-01-01

The production of a homogeneous protein sample in sufficient quantities is an essential prerequisite not only for structural investigations but represents also a rate-limiting step for many functional studies. In the cell, a large fraction of eukaryotic proteins exists as large multicomponent assemblies with many subunits, which act in concert to catalyze specific activities. Many of these complexes cannot be obtained from endogenous source material, so recombinant expression and reconstitution are then required to overcome this bottleneck. This chapter describes current strategies and protocols for the efficient production of multiprotein complexes in large quantities and of high quality, using the baculovirus/insect cell expression system.

High-throughput synchronization of mammalian cell cultures by spiral microfluidics.

PubMed

Lee, Wong Cheng; Bhagat, Ali Asgar S; Lim, Chwee Teck

2014-01-01

The development of mammalian cell cycle synchronization techniques has greatly advanced our understanding of many cellular regulatory events and mechanisms specific to different phases of the cell cycle. In this chapter, we describe a high-throughput microfluidic-based approach for cell cycle synchronization. By exploiting the relationship between cell size and its phase in the cell cycle, large numbers of synchronized cells can be obtained by size fractionation in a spiral microfluidic channel. Protocols for the synchronization of primary cells such as mesenchymal stem cells, and immortal cell lines such as Chinese hamster ovarian cells (CHO-CD36) and HeLa cells are provided as examples.

Metatranscriptomics of Soil Eukaryotic Communities.

PubMed

Yadav, Rajiv K; Bragalini, Claudia; Fraissinet-Tachet, Laurence; Marmeisse, Roland; Luis, Patricia

2016-01-01

Functions expressed by eukaryotic organisms in soil can be specifically studied by analyzing the pool of eukaryotic-specific polyadenylated mRNA directly extracted from environmental samples. In this chapter, we describe two alternative protocols for the extraction of high-quality RNA from soil samples. Total soil RNA or mRNA can be converted to cDNA for direct high-throughput sequencing. Polyadenylated mRNA-derived full-length cDNAs can also be cloned in expression plasmid vectors to constitute soil cDNA libraries, which can be subsequently screened for functional gene categories. Alternatively, the diversity of specific gene families can also be explored following cDNA sequence capture using exploratory oligonucleotide probes.

Sweet Potato [Ipomoea batatas (L.) Lam].

PubMed

Song, Guo-qing; Yamaguchi, Ken-ichi

2006-01-01

Among the available transformation methods reported on sweet potato, Agrobacterium tumefaciens-mediated transformation is more successful and desirable. Stem explants have shown to be ideal for the transformation of sweet potato because of their ready availability as explants, the simple transformation process, and high-frequency-regeneration via somatic embryogenesis. Under the two-step kanamycin-hygromycin selection method and using the appropriate explants type (stem explants), the efficiency of transformation can be considerably improved in cv. Beniazuma. The high efficiency in the transformation of stem explants suggests that the transformation protocol described in this chapter warrants testing for routine stable transformation of diverse varieties of sweet potato.

Software-Defined Architectures for Spectrally Efficient Cognitive Networking in Extreme Environments

NASA Astrophysics Data System (ADS)

Sklivanitis, Georgios

The objective of this dissertation is the design, development, and experimental evaluation of novel algorithms and reconfigurable radio architectures for spectrally efficient cognitive networking in terrestrial, airborne, and underwater environments. Next-generation wireless communication architectures and networking protocols that maximize spectrum utilization efficiency in congested/contested or low-spectral availability (extreme) communication environments can enable a rich body of applications with unprecedented societal impact. In recent years, underwater wireless networks have attracted significant attention for military and commercial applications including oceanographic data collection, disaster prevention, tactical surveillance, offshore exploration, and pollution monitoring. Unmanned aerial systems that are autonomously networked and fully mobile can assist humans in extreme or difficult-to-reach environments and provide cost-effective wireless connectivity for devices without infrastructure coverage. Cognitive radio (CR) has emerged as a promising technology to maximize spectral efficiency in dynamically changing communication environments by adaptively reconfiguring radio communication parameters. At the same time, the fast developing technology of software-defined radio (SDR) platforms has enabled hardware realization of cognitive radio algorithms for opportunistic spectrum access. However, existing algorithmic designs and protocols for shared spectrum access do not effectively capture the interdependencies between radio parameters at the physical (PHY), medium-access control (MAC), and network (NET) layers of the network protocol stack. In addition, existing off-the-shelf radio platforms and SDR programmable architectures are far from fulfilling runtime adaptation and reconfiguration across PHY, MAC, and NET layers. Spectrum allocation in cognitive networks with multi-hop communication requirements depends on the location, network traffic load, and interference profile at each network node. As a result, the development and implementation of algorithms and cross-layer reconfigurable radio platforms that can jointly treat space, time, and frequency as a unified resource to be dynamically optimized according to inter- and intra-network interference constraints is of fundamental importance. In the next chapters, we present novel algorithmic and software/hardware implementation developments toward the deployment of spectrally efficient terrestrial, airborne, and underwater wireless networks. In Chapter 1 we review the state-of-art in commercially available SDR platforms, describe their software and hardware capabilities, and classify them based on their ability to enable rapid prototyping and advance experimental research in wireless networks. Chapter 2 discusses system design and implementation details toward real-time evaluation of a software-radio platform for all-spectrum cognitive channelization in the presence of narrowband or wideband primary stations. All-spectrum channelization is achieved by designing maximum signal-to-interference-plus-noise ratio (SINR) waveforms that span the whole continuum of the device-accessible spectrum, while satisfying peak power and interference temperature (IT) constraints for the secondary and primary users, respectively. In Chapter 3, we introduce the concept of all-spectrum channelization based on max-SINR optimized sparse-binary waveforms, we propose optimal and suboptimal waveform design algorithms, and evaluate their SINR and bit-error-rate (BER) performance in an SDR testbed. Chapter 4 considers the problem of channel estimation with minimal pilot signaling in multi-cell multi-user multi-input multi-output (MIMO) systems with very large antenna arrays at the base station, and proposes a least-squares (LS)-type algorithm that iteratively extracts channel and data estimates from a short record of data measurements. Our algorithmic developments toward spectrally-efficient cognitive networking through joint optimization of channel access code-waveforms and routes in a multi-hop network are described in Chapter 5. Algorithmic designs are software optimized on heterogeneous multi-core general-purpose processor (GPP)-based SDR architectures by leveraging a novel software-radio framework that offers self-optimization and real-time adaptation capabilities at the PHY, MAC, and NET layers of the network protocol stack. Our system design approach is experimentally validated under realistic conditions in a large-scale hybrid ground-air testbed deployment. Chapter 6 reviews the state-of-art in software and hardware platforms for underwater wireless networking and proposes a software-defined acoustic modem prototype that enables (i) cognitive reconfiguration of PHY/MAC parameters, and (ii) cross-technology communication adaptation. The proposed modem design is evaluated in terms of effective communication data rate in both water tank and lake testbed setups. In Chapter 7, we present a novel receiver configuration for code-waveform-based multiple-access underwater communications. The proposed receiver is fully reconfigurable and executes (i) all-spectrum cognitive channelization, and (ii) combined synchronization, channel estimation, and demodulation. Experimental evaluation in terms of SINR and BER show that all-spectrum channelization is a powerful proposition for underwater communications. At the same time, the proposed receiver design can significantly enhance bandwidth utilization. Finally, in Chapter 8, we focus on challenging practical issues that arise in underwater acoustic sensor network setups where co-located multi-antenna sensor deployment is not feasible due to power, computation, and hardware limitations, and design, implement, and evaluate an underwater receiver structure that accounts for multiple carrier frequency and timing offsets in virtual (distributed) MIMO underwater systems.

Chapter 23: Combined Heat and Power Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W.; Simons, George; Barsun, Stephan

The main focus of most evaluations is to determine the energy-savings impacts of the installed measure. This protocol defines a combined heat and power (CHP) measure as a system that sequentially generates both electrical energy and useful thermal energy from one fuel source at a host customer's facility or residence. This protocol is aimed primarily at regulators and administrators of ratepayer-funded CHP programs; however, project developers may find the protocol useful to understand how CHP projects are evaluated.

[Conditioning mechanisms and psychoneuroimmunology].

PubMed

Stockhorst, Ursula; Klosterhalfen, Sibylle

2005-01-01

This chapter deals with the role of conditioning principles in psychoneuroimmunology (PNI). We will first describe the paradigms of classical and instrumental conditioning and classify immune parameters that are subject to conditioning (chapter 1). So far, PNI research mainly uses classical (or Pavlovian) conditioning. We will summarize some of the paradigmatic studies, mainly animal studies (chapter 2) and also describe studies that support the clinical relevance of classical conditioning, i. e., in the pharmacological treatment of autoimmune diseases, transplantation and tumor chemotherapy (chapter 3). A study of our group on anticipatory immunomodulation in pediatric cancer patients is reported. Mechanisms mediating conditioned immunomodulation are summarized (chapter 4). We also describe studies that analyze the impact of instrumental conditioning contingencies on immune functioning (chapter 5). Finally, research perspectives are summarized (chapter 6).

Friendly Neighborhood Computer Project. Extension of the IBM NJE network to DEC VAX computers

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

Raffenetti, R.C.; Bertoncini, P.J.; Engert, D.E.

1984-07-01

This manual is divided into six chapters. The first is an overview of the VAX NJE emulator system and describes what can be done with the VAX NJE emulator software. The second chapter describes the commands that users of the VAX systems will use. Each command description includes the format of the command, a list of valid options and parameters and their meanings, and several short examples of command use. The third chapter describes the commands and capabilities for sending general, sequential files from and to VAX VMS nodes. The fourth chapter describes how to transmit data to a VAXmore » from other computer systems on the network. The fifth chapter explains how to exchange electronic mail with IBM CMS users and with users of other VAX VMS systems connected by NJE communications. The sixth chapter describes operator procedures and the additional commands operators may use.« less

An Analysis of the Department of Defense’s Use of Other Transaction Authority (10 U.S.C. 2371)

DTIC Science & Technology

2016-06-01

and protocols and will also review the advantages / benefits , disadvantages /risks, and lessons learned with respect to OT authority. Finally, Chapter V...examined the various advantages / benefits and disadvantages /risks of OT authority. Finally, this Chapter provided a few examples of technologies that have... ADVANTAGES AND BENEFITS .........................................................44 1. Flexible Terms and Conditions

Vocational Education and Training in Norway.

ERIC Educational Resources Information Center

Farstad, Halfdan

This volume on Norway describes the country's initial and continuing vocational education and training (VET) system. Chapter 1 provides background information on political and administrative structures; population; and economy and labor force. Chapter 2 describes briefly the Norwegian education and training system. Chapter 3 describes the VET…

The Soviet System of Education. A PIER World Education Series Special Report.

ERIC Educational Resources Information Center

Popovych, Erika; Levin-Stankevich, Brian

This volume endeavors to provide comprehensive factual information on the Soviet system of education. Chapter 1 offers basic information on the Soviet Republics. Chapter 2 describes the foundations of Soviet Education. Chapter 3 describes preschool through upper secondary education including academic calendars and curriculum. Chapter 4,…

Progress in coherent lithography using table-top extreme ultraviolet lasers

NASA Astrophysics Data System (ADS)

Li, Wei

Nanotechnology has drawn a wide variety of attention as interesting phenomena occurs when the dimension of the structures is in the nanometer scale. The particular characteristics of nanoscale structures had enabled new applications in different fields in science and technology. Our capability to fabricate these nanostructures routinely for sure will impact the advancement of nanoscience. Apart from the high volume manufacturing in semiconductor industry, a small-scale but reliable nanofabrication tool can dramatically help the research in the field of nanotechnology. This dissertation describes alternative extreme ultraviolet (EUV) lithography techniques which combine table-top EUV laser and various cost-effective imaging strategies. For each technique, numerical simulations, system design, experiment result and its analysis will be presented. In chapter II, a brief review of the main characteristics of table-top EUV lasers will be addressed concentrating on its high power and large coherence radius that enable the lithography application described herein. The development of a Talbot EUV lithography system which is capable of printing 50nm half pitch nanopatterns will be illustrated in chapter III. A detailed discussion of its resolution limit will be presented followed by the development of X-Y-Z positioning stage, the fabrication protocol for diffractive EUV mask, and the pattern transfer using self- developed ion beam etching, and the dose control unit. In addition, this dissertation demonstrated the capability to fabricate functional periodic nanostructures using Talbot EUV lithography. After that, resolution enhancement techniques like multiple exposure, displacement Talbot EUV lithography, fractional Talbot EUV lithography, and Talbot lithography using 18.9nm amplified spontaneous emission laser will be demonstrated. Chapter IV will describe a hybrid EUV lithography which combines the Talbot imaging and interference lithography rendering a high resolution interference pattern whose lattice is modified by a custom designed Talbot mask. In other words, this method enables filling the arbitrary Talbot cell with ultra-fine interference nanofeatures. Detailed optics modeling, system design and experiment results using He-Ne laser and table top EUV laser are included. The last part of chapter IV will analyze its exclusive advantages over traditional Talbot or interference lithography.

Generation of Neural Progenitor Spheres from Human Pluripotent Stem Cells in a Suspension Bioreactor.

PubMed

Yan, Yuanwei; Song, Liqing; Tsai, Ang-Chen; Ma, Teng; Li, Yan

2016-01-01

Conventional two-dimensional (2-D) culture systems cannot provide large numbers of human pluripotent stem cells (hPSCs) and their derivatives that are demanded for commercial and clinical applications in in vitro drug screening, disease modeling, and potentially cell therapy. The technologies that support three-dimensional (3-D) suspension culture, such as a stirred bioreactor, are generally considered as promising approaches to produce the required cells. Recently, suspension bioreactors have also been used to generate mini-brain-like structure from hPSCs for disease modeling, showing the important role of bioreactor in stem cell culture. This chapter describes a detailed culture protocol for neural commitment of hPSCs into neural progenitor cell (NPC) spheres using a spinner bioreactor. The basic steps to prepare hPSCs for bioreactor inoculation are illustrated from cell thawing to cell propagation. The method for generating NPCs from hPSCs in the spinner bioreactor along with the static control is then described. The protocol in this study can be applied to the generation of NPCs from hPSCs for further neural subtype specification, 3-D neural tissue development, or potential preclinical studies or clinical applications in neurological diseases.

Faster experimental validation of microRNA targets using cold fusion cloning and a dual firefly-Renilla luciferase reporter assay.

PubMed

Alvarez, M Lucrecia

2014-01-01

Different target prediction algorithms have been developed to provide a list of candidate target genes for a given animal microRNAs (miRNAs). However, these computational approaches provide both false-positive and false-negative predictions. Therefore, the target genes of a specific miRNA identified in silico should be experimentally validated. In this chapter, we describe a step-by-step protocol for the experimental validation of a direct miRNA target using a faster Dual Firefly-Renilla Luciferase Reporter Assay. We describe how to construct reporter plasmids using the simple, fast, and highly efficient cold fusion cloning technology, which does not require ligase, phosphatase, or restriction enzymes. In addition, we provide a protocol for co-transfection of reporter plasmids with either miRNA mimics or miRNA inhibitors in human embryonic kidney 293 (HEK293) cells, as well as a description on how to measure Firefly and Renilla luciferase activity using the Dual-Glo Luciferase Assay kit. As an example of the use of this technology, we will validate glucose-6-phosphate dehydrogenase (G6PD) as a direct target of miR-1207-5p.

Chapter 12: Survey Design and Implementation for Estimating Gross Savings Cross-Cutting Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Baumgartner, Robert

This chapter presents an overview of best practices for designing and executing survey research to estimate gross energy savings in energy efficiency evaluations. A detailed description of the specific techniques and strategies for designing questions, implementing a survey, and analyzing and reporting the survey procedures and results is beyond the scope of this chapter. So for each topic covered below, readers are encouraged to consult articles and books cited in References, as well as other sources that cover the specific topics in greater depth. This chapter focuses on the use of survey methods to collect data for estimating gross savingsmore » from energy efficiency programs.« less

Composite material impregnation unit

NASA Technical Reports Server (NTRS)

Wilkinson, S. P.; Marchello, J. M.; Johnston, N. J.

1993-01-01

This memorandum presents an introduction to the NASA multi-purpose prepregging unit which is now installed and fully operational at the Langley Research Center in the Polymeric Materials Branch. A description of the various impregnation methods that are available to the prepregger are presented. Machine operating details and protocol are provided for its various modes of operation. These include, where appropriate, the related equations for predicting the desired prepreg specifications. Also, as the prepregger is modular in its construction, each individual section is described and discussed. Safety concerns are an important factor and a chapter has been included that highlights the major safety features. Initial experiences and observations for fiber impregnation are described. These first observations have given great insight into the areas of future work that need to be addressed. Future memorandums will focus on these individual processes and their related problems.

Chapter 14: Chiller Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Tiessen, Alex

This protocol defines a chiller measure as a project that directly impacts equipment within the boundary of a chiller plant. A chiller plant encompasses a chiller - or multiple chillers - and associated auxiliary equipment. This protocol primarily covers electric-driven chillers and chiller plants. It does not include thermal energy storage and absorption chillers fired by natural gas or steam, although a similar methodology may be applicable to these chilled water system components.

Production and Electrical Characterization Tests of the ISL Detector and a Trigger Design for Higgs Boson Searches at CDF

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

Munar Ara, Antoni

2002-01-01

This thesis is structured as follows: Chapter 1. gives a brief review of the Higgs mechanism in the Standard Model and the electroweak symmetry breaking. The Standard Model Higgs boson phenomenology at Tevatron energies is reviewed. Chapter 2. describes the upgraded Fermilab laboratory accelerator complex, and the upgraded CDF detector. Chapter 3. gives a brief overview of the more relevant aspects of the silicon detectors, and the ISL is described in detail. Chapter 4. describes the construction of the ISL ladders, the full custom testing setup (functionality tests, laser test, burn-in test andmore » $$\\beta$$-source measurements), and the problems encountered during the ISL ladders construction. The procedures for ladder grading are also discussed. Chapter 5. describes the multilevel trigger system of the CDF detector, and the trigger primitives available at each level. The most relevant offine event observables are briefly discussed. In Chapter 6 the procedures to estimate the trigger rate and trigger effciency calculation are described. The particularities of triggering in $$p\\bar{p}$$ collisions at high luminosities are discussed. Chapter 7. and Chapter 8. are dedicated to study an effcient trigger strategy for the $$H + W/Z \\to b\\bar{b}jj$$ channel and the $$H + Z \\to b\\bar{b} \

Molecular Dynamics Simulation and Analysis of the Antimicrobial Peptide-Lipid Bilayer Interactions.

PubMed

Arasteh, Shima; Bagheri, Mojtaba

2017-01-01

A great deal of research has been undertaken in order to discover antimicrobial peptides (AMPs) with unexploited mechanisms of action to counteract the health-threatening issues associated with bacterial resistance. The intrinsic effectiveness of AMPs is strongly influenced by their initial interactions with the bacterial cell membrane. Understanding these interactions in the atomistic details is important for the design of the less prone bacteria-resistant peptides. However, these studies always require labor-intensive and difficult steps. With this regard, modeling studies of the AMPs binding to simple lipid membrane systems, e.g., lipid bilayers, is of great advantage. In this chapter, we present an applicable step-by-step protocol to run the molecular dynamics (MD) simulation of the interaction between cyclo-RRWFWR (c-WFW) (a small cyclic AMP) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid bilayer using the Groningen machine for chemical simulations (GROMACS) package. The protocol as described here may simply be optimized for other peptide-lipid systems of interest.

All-in-One CRISPR-Cas9/FokI-dCas9 Vector-Mediated Multiplex Genome Engineering in Cultured Cells.

PubMed

Sakuma, Tetsushi; Sakamoto, Takuya; Yamamoto, Takashi

2017-01-01

CRISPR-Cas9 enables highly convenient multiplex genome engineering in cultured cells, because it utilizes generic Cas9 nuclease and an easily customizable single-guide RNA (sgRNA) for site-specific DNA double-strand break induction. We previously established a multiplex CRISPR-Cas9 assembly system for constructing an all-in-one vector simultaneously expressing multiple sgRNAs and Cas9 nuclease or other Cas9 variants including FokI-dCas9, which supersedes the wild-type Cas9 with regard to high specificity. In this chapter, we describe a streamlined protocol to design and construct multiplex CRISPR-Cas9 or FokI-dCas9 vectors, to introduce them into cultured cells by lipofection or electroporation, to enrich the genomically edited cells with a transient puromycin selection, to validate the mutation efficiency by Surveyor nuclease assay, and to perform off-target analyses. We show that our protocol enables highly efficient multiplex genome engineering even in hard-to-transfect HepG2 cells.

Cultivation, detection, and ecophysiology of anaerobic ammonium-oxidizing bacteria.

PubMed

Kartal, Boran; Geerts, Wim; Jetten, Mike S M

2011-01-01

Anaerobic ammonium-oxidizing (anammox) bacteria oxidize ammonium with nitrite under anoxic conditions. The anammox process is currently used to remove ammonium from wastewater and contributes significantly to the loss of fixed nitrogen from the oceans. In this chapter, we focus on the ecophysiology of anammox bacteria and describe new methodologies to grow these microorganisms. Now, it is possible to enrich anammox bacteria up to 95% with a membrane bioreactor that removes forces of selection for fast settling aggregates and facilitates the growth of planktonic cells. The biomass from this system has a high anaerobic ammonium oxidation rate (50 fmol NH(4)(+) · cell(-1) day(-1)) and is suitable for many ecophysiological and molecular experiments. A high throughput Percoll density gradient centrifugation protocol may be applied on this biomass for further enrichment (>99.5%) of anammox bacteria. Furthermore, we provide an up-to-date list of commonly used primers and introduce protocols for quantification and detection of functional genes of anammox bacteria in their natural environment. Copyright © 2011 Elsevier Inc. All rights reserved.

Bermudagrass (Cynodon spp.).

PubMed

Ge, Yaxin; Wang, Zeng-Yu

2006-01-01

Bermudagrass is an important warm-season forage and turf species widely grown in the southern United States. This chapter describes a rapid and efficient protocol that allows for the generation of a large number of transgenic bermudagrass plants, bypassing the callus formation phase. Stolon nodes are infected and co-cultivated with Agrobacterium tumefaciens harboring pCAMBIA binary vectors. Hygromycin phosphotransferase gene (hph) is used as the selectable marker and hygromycin is used as the selection agent. Green shoots are directly produced from infected stolon nodes 4 to 5 wk after hygromycin selection. Without callus formation and with minimum tissue culture, this procedure allowed us to obtain well-rooted transgenic plantlets in only 7 wk and greenhouse-grown plants in only 9 wk.

Proteomic Profiling of the Pituitary Gland in Studies of Psychiatric Disorders.

PubMed

Krishnamurthy, Divya; Rahmoune, Hassan; Guest, Paul C

2017-01-01

Psychiatric disorders have been associated with perturbations of the hypothalamic-pituitary-adrenal axis. Therefore, proteomic studies of the pituitary gland have the potential to provide new insights into the underlying pathways affected in these conditions as well as identify new biomarkers or targets for use in developing improved medications. This chapter describes a protocol for preparation of pituitary protein extracts followed by characterization of the pituitary proteome by label-free liquid chromatography-tandem mass spectrometry in expression mode (LC-MS E ). The main focus was on establishing a method for identifying the major pituitary hormones and accessory proteins as many of these have already been implicated in psychiatric diseases.

Micropropagation and in vitro conservation of vanilla (Vanilla planifolia Andrews).

PubMed

Divakaran, Minoo; Babu, K Nirmal

2009-01-01

Vanilla (Vanilla planifolia Andrews (syn. V. fragrans Salisb.), a source of natural vanillin, plays a major positive role in the economy of several countries. A native to the Central America, its primary gene pool is threatened by deforestation and over collection that has resulted in disappearance of natural habitats and wild species. Therefore, multiplication and conservation of vanilla diversity is of paramount importance because of its narrow genetic base. It plays an important role in the production of disease free planting material for commercial cultivation. Simple protocols for micropropagation, in vitro conservation and synthetic seed production are described in this chapter which could further be applied to other related vanilla species as well.

Petunia (Petunia hybrida).

PubMed

Lutke, W Kevin

2006-01-01

Petunia hybrida genetic transformation continues to be a valuable tool for genetic research into biochemical pathways and gene expression, as well as generating commercial products with varying floral colors. In this chapter, we describe a simple and reproducible genetic transformation protocol for generating transgenic petunia plants harboring a gene of interest and selectable marker. The system utilizes Agrobacterium tumefaciens for transgene integration with plant recovery via shoot organogenesis from leaf explant material. Selection for transgenic plants is achieved using the bar gene conferring resistance to glufosinate or nptII gene for resistance to kanamycin. Transformation efficiencies of around 10% are achievable with shoots being recovered about 8 wk after transgene insertion and rooted plants transferred to the greenhouse about twelve weeks after inoculation.

Analysis of ChIP-seq Data in R/Bioconductor.

PubMed

de Santiago, Ines; Carroll, Thomas

2018-01-01

The development of novel high-throughput sequencing methods for ChIP (chromatin immunoprecipitation) has provided a very powerful tool to study gene regulation in multiple conditions at unprecedented resolution and scale. Proactive quality-control and appropriate data analysis techniques are of critical importance to extract the most meaningful results from the data. Over the last years, an array of R/Bioconductor tools has been developed allowing researchers to process and analyze ChIP-seq data. This chapter provides an overview of the methods available to analyze ChIP-seq data based primarily on software packages from the open-source Bioconductor project. Protocols described in this chapter cover basic steps including data alignment, peak calling, quality control and data visualization, as well as more complex methods such as the identification of differentially bound regions and functional analyses to annotate regulatory regions. The steps in the data analysis process were demonstrated on publicly available data sets and will serve as a demonstration of the computational procedures routinely used for the analysis of ChIP-seq data in R/Bioconductor, from which readers can construct their own analysis pipelines.

Chapter 13: Assessing Persistence and Other Evaluation Issues Cross-Cutting Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Violette, Daniel M.

Addressing other evaluation issues that have been raised in the context of energy efficiency programs, this chapter focuses on methods used to address the persistence of energy savings, which is an important input to the benefit/cost analysis of energy efficiency programs and portfolios. In addition to discussing 'persistence' (which refers to the stream of benefits over time from an energy efficiency measure or program), this chapter provides a summary treatment of these issues -Synergies across programs -Rebound -Dual baselines -Errors in variables (the measurement and/or accuracy of input variables to the evaluation).

Ecological Census Techniques - 2nd Edition

NASA Astrophysics Data System (ADS)

Sutherland, Edited By William J.

2006-08-01

This is an updated version of the best selling first edition, Ecological Census Techniques, with updating, some new chapters and authors. Almost all ecological and conservation work involves carrying out a census or survey. This practically focussed book describes how to plan a census, the practical details and shows with worked examples how to analyse the results. The first three chapters describe planning, sampling and the basic theory necessary for carrying out a census. In the subsequent chapters international experts describe the appropriate methods for counting plants, insects, fish, amphibians, reptiles, mammals and birds. As many censuses also relate the results to environmental variability, there is a chapter explaining the main methods. Finally, there is a list of the most common mistakes encountered when carrying out a census. Gives worked examples and describes practical details The chapter on research planning provides an approach for planning any research, not just those relating to census techniques Latest edition of a very highly-regarded book. Includes new authors, each chapter has been updated, and additional chapters on sampling and designing research programmes have been added

Expression and analysis of exogenous proteins in epidermal cells.

PubMed

Dagnino, Lina; Ho, Ernest; Chang, Wing Y

2010-01-01

In this chapter we review protocols for transient transfection of primary keratinocytes. The ability to transfect primary epidermal cells regardless of their differentiation status allows the biochemical and molecular characterization of multiple proteins. We review methods to analyze exogenous protein abundance in transfected keratinocytes by immunoblot and immunoprecipitation. We also present protocols to determine the subcellular distribution of these proteins by indirect immunofluorescence microscopy approaches.

Telemetry Standards, RCC Standard 106-17. Chapter 26. TmNSDataMessage Transfer Protocol

DTIC Science & Technology

2017-07-01

Channel (RTSPDataChannel) ............................................ 26-13 26.4.3 Reliability Critical (RC) Delivery Protocol...error status code specified in RFC 2326 for "Request-URI Too Large" is 虮". 26.4.1.5 Request Types RTSPDataSources shall return valid ...to the following requirements. • Valid TmNSDataMessages shall be delivered containing the original Packages matching the requested

The Training/Development of In-Company Trainers/Supervisors of Young People--The Case of Greece.

ERIC Educational Resources Information Center

Constantinides, Xenophon; And Others

This document describes the training of the in-company trainer of young people in Greece. Chapter 1 describes the context of training. Chapter 2 describes the methodology used in the study including questionnaires, interviews, and a forum. Chapter 3 reports the results of the study in the following categories: training manager, in-company trainer,…

Explanatory chapter: how plasmid preparation kits work.

PubMed

Koontz, Laura

2013-01-01

To isolate plasmid DNA from bacteria using a commercial plasmid miniprep kit (if interested, compare this protocol with Isolation of plasmid DNA from bacteria). Copyright © 2013 Elsevier Inc. All rights reserved.

BAC sequencing using pooled methods.

PubMed

Saski, Christopher A; Feltus, F Alex; Parida, Laxmi; Haiminen, Niina

2015-01-01

Shotgun sequencing and assembly of a large, complex genome can be both expensive and challenging to accurately reconstruct the true genome sequence. Repetitive DNA arrays, paralogous sequences, polyploidy, and heterozygosity are main factors that plague de novo genome sequencing projects that typically result in highly fragmented assemblies and are difficult to extract biological meaning. Targeted, sub-genomic sequencing offers complexity reduction by removing distal segments of the genome and a systematic mechanism for exploring prioritized genomic content through BAC sequencing. If one isolates and sequences the genome fraction that encodes the relevant biological information, then it is possible to reduce overall sequencing costs and efforts that target a genomic segment. This chapter describes the sub-genome assembly protocol for an organism based upon a BAC tiling path derived from a genome-scale physical map or from fine mapping using BACs to target sub-genomic regions. Methods that are described include BAC isolation and mapping, DNA sequencing, and sequence assembly.

Acute and Chronic Exercise in Animal Models.

PubMed

Thu, Vu Thi; Kim, Hyoung Kyu; Han, Jin

2017-01-01

Numerous animal cardiac exercise models using animal subjects have been established to uncover the cardiovascular physiological mechanism of exercise or to determine the effects of exercise on cardiovascular health and disease. In most cases, animal-based cardiovascular exercise modalities include treadmill running, swimming, and voluntary wheel running with a series of intensities, times, and durations. Those used animals include small rodents (e.g., mice and rats) and large animals (e.g., rabbits, dogs, goats, sheep, pigs, and horses). Depending on the research goal, each experimental protocol should also describe whether its respective exercise treatment can produce the anticipated acute or chronic cardiovascular adaptive response. In this chapter, we will briefly describe the most common kinds of animal models of acute and chronic cardiovascular exercises that are currently being conducted and are likely to be chosen in the near future. Strengths and weakness of animal-based cardiac exercise modalities are also discussed.

In Vitro and Cryopreservation Techniques for Conservation of Snow Mountain Garlic.

PubMed

Mahajan, Ritu

2016-01-01

Garlic is an important medicinal herb of culinary value by imparting its flavors and odors to the food. Allicin, a notable flavonoid in garlic, is a powerful antibiotic and antifungal compound. Due to poor bioavailability, garlic is of limited use for oral human consumption. Being sexually sterile, propagation of garlic is done by individual cloves from a bulb which increases the chances of transfer of viral diseases. In this chapter, an efficient and improved regeneration protocol for explant establishment and shoot multiplication under in vitro conditions is described. A high rate of shoot multiplication is obtained on MS medium supplemented with 0.5 mg/l BAP, 1.0 mg/l KN, and 2.0 mg/l GA3. Addition of 1.0 mg/l NAA to MS medium resulted in rooting at the shoot bases. A detailed method for encapsulation of explant in sodium alginate beads and their cryopreservation using encapsulation-dehydration is also described.

Characterization of a Protein Interactome by Co-Immunoprecipitation and Shotgun Mass Spectrometry.

PubMed

Maccarrone, Giuseppina; Bonfiglio, Juan Jose; Silberstein, Susana; Turck, Christoph W; Martins-de-Souza, Daniel

2017-01-01

Identifying the partners of a given protein (the interactome) may provide leads about the protein's function and the molecular mechanisms in which it is involved. One of the alternative strategies used to characterize protein interactomes consists of co-immunoprecipitation (co-IP) followed by shotgun mass spectrometry. This enables the isolation and identification of a protein target in its native state and its interactome from cells or tissue lysates under physiological conditions. In this chapter, we describe a co-IP protocol for interactome studies that uses an antibody against a protein of interest bound to protein A/G plus agarose beads to isolate a protein complex. The interacting proteins may be further fractionated by SDS-PAGE, followed by in-gel tryptic digestion and nano liquid chromatography high-resolution tandem mass spectrometry (nLC ESI-MS/MS) for identification purposes. The computational tools, strategy for protein identification, and use of interactome databases also will be described.

UNDERSTANDING AND ACCOUNTING FOR METHOD VARIABILITY IN WHOLE EFFLUENT TOXICITY APPLICATIONS UNDER THE NPDES PROGRAM

EPA Science Inventory

This chapter provides a brief introduction to whole effluent toxicity (WET) testing and describes the regulatory background and context of WET testing. This chapter also describes the purpose of this document and outlines the issues addressed in each chapter.

Stanford Synchrotron Radiation Laboratory activity report for 1986

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

Cantwell, K.

1987-12-31

1986 was another year of major advances for SSRL as the ultimate capabilities of PEP as a synchrotron radiation source became more apparent and a second PEP beam line was initiated, while effective development and utilization of SPEAR proceeded. Given these various PEP developments, SSRL abandoned its plans for a separate diffraction limited ring, as they abandoned their plans for a 6--7 GeV ring of the APS type last year. It has become increasingly apparent that SSRL should concentrate on developing SPEAR and PEP as synchrotron radiation sources. Consequently, initial planning for a 3 GeV booster synchrotron injector for SPEARmore » was performed in 1986, with a proposal to the Department of Energy resulting. As described in Chapter 2, the New Rings Group and the Machine Physics Group were combined into one Accelerator Physics Group. This group is focusing mainly on the improvement of SPEAR`s operating conditions and on planning for the conversion of PEP into a fourth generation x-ray source. Considerable emphasis is also being given to the training of accelerator physics graduate students. At the same time, several improvements of SSRL`s existing facilities were made. These are described in Chapter 3. Chapter 4 describes new SSRL beam lines being commissioned. Chapter 5 discusses SSRL`s present construction projects. Chapter 6 discusses a number of projects presently underway in the engineering division. Chapter 7 describes SSRL`s advisory panels while Chapter 8 discusses SSRL`s overall organization. Chapter 9 describes the experimental progress reports.« less

A measurement of the mass of the top quark using the ideogram technique

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

Houben, Pieter Willem Huib

2009-06-03

This thesis describes a measurement of the mass of the top quark on data collected with the D0 detector at the Tevatron collider in the period from 2002 until 2006. The first chapter describes the Standard Model and the prominent role of the top quark mass. The second chapter gives a description of the D0 detector which is used for this measurement. After the pmore » $$\\bar{p}$$ collisions have been recorded, reconstruction of physics objects is required, which is described in Chapter 3. Chapter 4 describes how the interesting collisions in which top quarks are produced are separated from the `uninteresting' ones with a set of selection criteria. The method to extract the top quark mass from the sample of selected collisions (also called events), which is based on the ideogram technique, is explained in Chapter 5, followed in Chapter 6 by the description of the calibration of the method using simulation of our most precise knowledge of nature. Chapter 7 shows the result of the measurement together with some cross checks and an estimation of the uncertainty on this measurement. This thesis concludes with a constraint on the Higgs boson mass.« less

The Monitoring and Affinity Purification of Proteins Using Dual Tags with Tetracysteine Motifs

NASA Astrophysics Data System (ADS)

Giannone, Richard J.; Liu, Yie; Wang, Yisong

Identification and characterization of protein-protein interaction networks is essential for the elucidation of biochemical mechanisms and cellular function. Affinity purification in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a very powerful tactic for the identification of specific protein-protein interactions. In this chapter, we describe a comprehensive methodology that uses our recently developed dual-tag affinity purification system for the enrichment and identification of mammalian protein complexes. The protocol covers a series of separate but sequentially related techniques focused on the facile monitoring and purification of a dual-tagged protein of interest and its interacting partners via a system built with tetracysteine motifs and various combinations of affinity tags. Using human telomeric repeat binding factor 2 (TRF2) as an example, we demonstrate the power of the system in terms of bait protein recovery after dual-tag affinity purification, detection of bait protein subcellular localization and expression, and successful identification of known and potentially novel TRF2 interacting proteins. Although the protocol described here has been optimized for the identification and characterization of TRF2-associated proteins, it is, in principle, applicable to the study of any other mammalian protein complexes that may be of interest to the research community.

Modeling habitat and environmental factors affecting mosquito abundance in Chesapeake, Virginia

NASA Astrophysics Data System (ADS)

Bellows, Alan Scott

The models I present in this dissertation were designed to enable mosquito control agencies in the mid-Atlantic region that oversee large jurisdictions to rapidly track the spatial and temporal distributions of mosquito species, especially those species known to be vectors of eastern equine encephalitis and West Nile virus. I was able to keep these models streamlined, user-friendly, and not cost-prohibitive using empirically based digital data to analyze mosquito-abundance patterns in real landscapes. This research is presented in three major chapters: (II) a series of semi-static habitat suitability indices (HSI) grounded on well-documented associations between mosquito abundance and environmental variables, (III) a dynamic model for predicting both spatial and temporal mosquito abundance based on a topographic soil moisture index and recent weather patterns, and (IV) a set of protocols laid out to aid mosquito control agencies for the use of these models. The HSIs (Chapter II) were based on relationships of mosquitoes to digital surrogates of soil moisture and vegetation characteristics. These models grouped mosquitoes species derived from similarities in habitat requirements, life-cycle type, and vector competence. Quantification of relationships was determined using multiple linear regression models. As in Chapter II, relationships between mosquito abundance and environmental factors in Chapter III were quantified using regression models. However, because this model was, in part, a function of changes in weather patterns, it enables the prediction of both 'where' and 'when' mosquito outbreaks are likely to occur. This model is distinctive among similar studies in the literature because of my use of NOAA's NEXRAD Doppler radar (3-hr precipitation accumulation data) to quantify the spatial and temporal distributions in precipitation accumulation. \\ Chapter IV is unique among the chapters in this dissertation because in lieu of presenting new research, it summarizes the preprocessing steps and analyses used in the HSIs and the dynamic, weather-based, model generated in Chapters II and III. The purpose of this chapter is to provide the reader and potential users with the necessary protocols for modeling the spatial and temporal abundances and distributions of mosquitoes, with emphasis on Culiseta melanura, in a real-world landscape of the mid-Atlantic region. This chapter also provides enhancements that could easily be incorporated into an environmentally sensitive integrated pest management program.

Synthesis and characterization of transition metal clusters: From the isolation of ligand-stabilized solid fragments to the tuning of magnetic anisotropy and host-guest selectivity, and, Approaches to science teaching: Development of an observation instrument with a measurement model based on item response theory

NASA Astrophysics Data System (ADS)

Hee, Allan George

Part I. The work presented herein describes efforts to develop general techniques for the synthesis of transition metal clusters and the manipulation of their properties. In Chapter 2, it is demonstrated that a modified metal atom reactor allows for the vaporization, passivation, and isolation of metal-chalcogenide clusters from their parent binary solids. Among the clusters produced by this method were Cr6S8(PEt3)6, Fe4S 4(PEt3)4, Co6S8(PEt 3)6, Cu6S4(PEt3)6, Cu12S6(PEt3)8, and Cu26Se 13(PEt3)14. To create single-molecule magnets with higher demagnetization barriers, we are developing metal-cyanide systems which exhibit highly adjustable magnetic behavior. Chapter 3 reports an attempt to introduce magnetic anisotropy into a MnCr6 cluster. Replacement of CrIII with Mo III resulted in the assembly of K[(Me3tacn)6MnMo 6(CN)18](ClO4)3 (Me3tacn = N,N',N″ -trimethyl-1,4,7-triazacyclononane)---the first well-documented example of a cyano-bridged single-molecule magnet. Recently, it was demonstrated that replacing Me3tacn with the less sterically hindering tach (tach = cis,cis-1,3,5-triaminocyclohexane) in the face-centered cubic cluster [(tach)8Cr8Ni 6(CN)24]Br12 provides greater access to the cluster cavity. Chapter 4 describes my efforts to probe the selectivity of this cluster toward inclusion of various guests. Part II. Successful implementation of student-centered curricula reforms requires the creation of a measurement instrument for monitoring whether the curricula are being used as intended. The creation and development of an observation instrument would greatly contribute to this effort. To develop a theoretically sound construct map, it is necessary to review the literature and conduct our own investigations of approaches to science teaching. Chapter 2 presents the findings of these investigations and their contributions to our understanding of the construct. Using these findings, the Science Teaching Observation Protocol (STOP) was created and designed to measure two subconstructs: intentions and strategies. Chapter 3 details the first pilot test of STOP and analysis of the collected data. In Chapter 4, the theoretical shortcomings of the instrument are analyzed and discussed. Modified versions of the intention and strategy subconstruct maps are presented.

Conclusion: From describing to prescribing--transitioning to place-based conservation [Chapter 18

Treesearch

William P. Stewart; Daniel R. Williams; Linda E. Kruger

2013-01-01

The chapters of this book describe various perspectives from the social sciences of place-based conservation. The prescriptive implications are often close to the surface and become entangled with them. This chapter highlights four overlapping approaches to the practice of place-based conservation and acknowledges the difficulty of separating descriptions from...

Chapter 6. Landscape Analysis for Habitat Monitoring

Treesearch

Samuel A. Cushman; Kevin McGarigal; Kevin S. McKelvey; Christina D. Vojta; Claudia M. Regan

2013-01-01

The primary objective of this chapter is to describe standardized methods for measur¬ing and monitoring attributes of landscape pattern in support of habitat monitoring. This chapter describes the process of monitoring categorical landscape maps in which either selected habitat attributes or different classes of habitat quality are represented as different patch types...

The Adult Basic Education Profession and Competence: Promoting Best Practice. Final Report 1993.

ERIC Educational Resources Information Center

Scheeres, Hermine; And Others

This report describes the adult basic education profession in Australia across state boundaries and across a wide range of contexts. Chapter 1 discusses competencies and professions and why competency-based teacher education has been unsuccessful. Chapter 2 discusses the context of Adult Basic Education Competencies. Chapter 3 describes the…

Chapter 08: Comments on, and additional information for, wood identification

Treesearch

Alex C. Wiedenhoeft

2011-01-01

This manual has described the theory of identification (Chapter 1), the botanical basis of wood structure (Chapter 2), the use of a hand lens (Chapter 3), how to use cutting tools to prepare wood for observation with a lens (Chapter 4), and the characters used in hand lens wood identification (Chapter 5) before leading you through an identification key (Chapter 6) and...

In vivo phosphorylation of WRKY transcription factor by MAPK.

PubMed

Ishihama, Nobuaki; Adachi, Hiroaki; Yoshioka, Miki; Yoshioka, Hirofumi

2014-01-01

Plants activate signaling networks in response to diverse pathogen-derived signals, facilitating transcriptional reprogramming through mitogen-activated protein kinase (MAPK) cascades. Identification of phosphorylation targets of MAPK and in vivo detection of the phosphorylated substrates are important processes to elucidate the signaling pathway in plant immune responses. We have identified a WRKY transcription factor, which is phosphorylated by defense-related MAPKs, SIPK and WIPK. Recent evidence demonstrated that some group I WRKY transcription factors, which contain a conserved motif in the N-terminal region, are activated by MAPK-dependent phosphorylation. In this chapter, we describe protocols for preparation of anti-phosphopeptide antibodies, detection of activated MAPKs using anti-phospho-MAPK antibody, and activated WRKY using anti-phospho-WRKY antibody, respectively.

The Controlled Cortical Impact Model of Experimental Brain Trauma: Overview, Research Applications, and Protocol

PubMed Central

Osier, Nicole; Dixon, C. Edward

2017-01-01

Controlled cortical impact (CCI) is a commonly used and highly regarded model of brain trauma that uses a pneumatically or electromagnetically controlled piston to induce reproducible and well-controlled injury. The CCI model was originally used in ferrets and it has since been scaled for use in many other species. This chapter will describe the historical development of the CCI model, compare and contrast the pneumatic and electromagnetic models, and summarize key short- and long-term consequences of TBI that have been gleaned using this model. In accordance with the recent efforts to promote high-quality evidence through the reporting of common data elements (CDEs), relevant study details—that should be reported in CCI studies—will be noted. PMID:27604719

Design and construction of functional AAV vectors.

PubMed

Gray, John T; Zolotukhin, Serge

2011-01-01

Using the basic principles of molecular biology and laboratory techniques presented in this chapter, researchers should be able to create a wide variety of AAV vectors for both clinical and basic research applications. Basic vector design concepts are covered for both protein coding gene expression and small non-coding RNA gene expression cassettes. AAV plasmid vector backbones (available via AddGene) are described, along with critical sequence details for a variety of modular expression components that can be inserted as needed for specific applications. Protocols are provided for assembling the various DNA components into AAV vector plasmids in Escherichia coli, as well as for transferring these vector sequences into baculovirus genomes for large-scale production of AAV in the insect cell production system.

Genome Sequencing of Steroid Producing Bacteria Using Ion Torrent Technology and a Reference Genome.

PubMed

Sola-Landa, Alberto; Rodríguez-García, Antonio; Barreiro, Carlos; Pérez-Redondo, Rosario

2017-01-01

The Next-Generation Sequencing technology has enormously eased the bacterial genome sequencing and several tens of thousands of genomes have been sequenced during the last 10 years. Most of the genome projects are published as draft version, however, for certain applications the complete genome sequence is required.In this chapter, we describe the strategy that allowed the complete genome sequencing of Mycobacterium neoaurum NRRL B-3805, an industrial strain exploited for steroid production, using Ion Torrent sequencing reads and the genome of a close strain as the reference. This protocol can be applied to analyze the genetic variations between closely related strains; for example, to elucidate the point mutations between a parental strain and a random mutagenesis-derived mutant.

Size analysis of polyglutamine protein aggregates using fluorescence detection in an analytical ultracentrifuge.

PubMed

Polling, Saskia; Hatters, Danny M; Mok, Yee-Foong

2013-01-01

Defining the aggregation process of proteins formed by poly-amino acid repeats in cells remains a challenging task due to a lack of robust techniques for their isolation and quantitation. Sedimentation velocity methodology using fluorescence detected analytical ultracentrifugation is one approach that can offer significant insight into aggregation formation and kinetics. While this technique has traditionally been used with purified proteins, it is now possible for substantial information to be collected with studies using cell lysates expressing a GFP-tagged protein of interest. In this chapter, we describe protocols for sample preparation and setting up the fluorescence detection system in an analytical ultracentrifuge to perform sedimentation velocity experiments on cell lysates containing aggregates formed by poly-amino acid repeat proteins.

Monitoring Phosphatidic Acid Signaling in Breast Cancer Cells Using Genetically Encoded Biosensors.

PubMed

Lu, Maryia; Tay, Li Wei Rachel; He, Jingquan; Du, Guangwei

2016-01-01

Phospholipids are important signaling molecules that regulate cell proliferation, death, migration, and metabolism. Many phospholipid signaling cascades are altered in breast cancer. To understand the functions of phospholipid signaling molecules, genetically encoded phospholipid biosensors have been developed to monitor their spatiotemporal dynamics. Compared to other phospholipids, much less is known about the subcellular production and cellular functions of phosphatidic acid (PA), partially due to the lack of a specific and sensitive PA biosensor in the past. This chapter describes the use of a newly developed PA biosensor, PASS, in two applications: regular fluorescent microscopy and fluorescence lifetime imaging microscopy-Förster/fluorescence resonance energy transfer (FLIM-FRET). These protocols can be also used with other phospholipid biosensors.

Murine macrophages: a technical approach.

PubMed

Martinez-Pomares, Luisa; Gordon, Siamon

2008-01-01

In this chapter, we describe current protocols used for the characterization of macrophages (MPhi) in mouse tissues and in cell suspensions from spleen and lymph nodes. Also, we include a brief description of a complementary approach: culture of primary MPhi. Although culture MPhi are extremely useful for analysing the basic biology of MPhi and their receptors, it should not be forgotten that the term MPhi encompasses a wide range of different types of cells with phenotypic characteristics dependent on their activation state and tissue of origin. In our view, there is no perfect MPhi marker and analysis of the expression profile of several markers, and functional studies are required to make an informed guess of the cellular characteristics and function of the MPhi population of interest.

Chapter A6. Section 6.6. Alkalinity and Acid Neutralizing Capacity

USGS Publications Warehouse

Rounds, Stewart A.; Wilde, Franceska D.

2002-01-01

Alkalinity (determined on a filtered sample) and Acid Neutralizing Capacity (ANC) (determined on a whole-water sample) are measures of the ability of a water sample to neutralize strong acid. Alkalinity and ANC provide information on the suitability of water for uses such as irrigation, determining the efficiency of wastewater processes, determining the presence of contamination by anthropogenic wastes, and maintaining ecosystem health. In addition, alkalinity is used to gain insights on the chemical evolution of an aqueous system. This section of the National Field Manual (NFM) describes the USGS field protocols for alkalinity/ANC determination using either the inflection-point or Gran function plot methods, including calculation of carbonate species, and provides guidance on equipment selection.

Isolation and Differentiation of Murine Macrophages.

PubMed

Rios, Francisco J; Touyz, Rhian M; Montezano, Augusto C

2017-01-01

Macrophages play a major role in inflammation, wound healing, and tissue repair. Infiltrated monocytes differentiate into different macrophage subtypes with protective or pathogenic activities in vascular lesions. In the heart and vascular tissues, pathological activation promotes cardiovascular inflammation and remodeling and there is increasing evidence that macrophages play important mechanisms in this environment. Primary murine macrophages can be obtained from: bone marrow by different treatments (granulocyte-macrophage colony-stimulating factor-GM-CSF, macrophage colony-stimulating factor-M-CSF or supernatant of murine fibroblast L929), peritoneal cavity (resident or thioglycolate elicit macrophages), from the lung (alveolar macrophages) or from adipose tissue. In this chapter we describe some protocols to obtain primary murine macrophages and how to identify a pure macrophage population or activation phenotypes using different markers.

Study of ATM Phosphorylation by Cdk5 in Neuronal Cells.

PubMed

She, Hua; Mao, Zixu

2017-01-01

The phosphatidylinositol-3-kinase-like kinase ATM (ataxia-telangiectasia mutated) plays a central role in coordinating the DNA damage responses including cell cycle checkpoint control, DNA repair, and apoptosis. Mutations of ATM cause a spectrum of defects ranging from neurodegeneration to cancer predisposition. We previously showed that Cdk5 (cyclin-dependent kinase 5) is activated by DNA damage and directly phosphorylates ATM at serine 794 in postmitotic neurons. Phosphorylation at serine 794 precedes and is required for ATM autophosphorylation at serine 1981, and activates ATM kinase activity. Cdk5-ATM pathway plays a crucial role in DNA damage-induced neuronal injury. This chapter describes protocols used in analyzing ATM phosphorylation by Cdk5 in CGNs (cerebellar granule neurons) and its effects on neuronal survival.

The Controlled Cortical Impact Model of Experimental Brain Trauma: Overview, Research Applications, and Protocol.

PubMed

Osier, Nicole; Dixon, C Edward

2016-01-01

Controlled cortical impact (CCI) is a commonly used and highly regarded model of brain trauma that uses a pneumatically or electromagnetically controlled piston to induce reproducible and well-controlled injury. The CCI model was originally used in ferrets and it has since been scaled for use in many other species. This chapter will describe the historical development of the CCI model, compare and contrast the pneumatic and electromagnetic models, and summarize key short- and long-term consequences of TBI that have been gleaned using this model. In accordance with the recent efforts to promote high-quality evidence through the reporting of common data elements (CDEs), relevant study details-that should be reported in CCI studies-will be noted.

Chapter 18: Variable Frequency Drive Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Romberger, Jeff

An adjustable-speed drive (ASD) includes all devices that vary the speed of a rotating load, including those that vary the motor speed and linkage devices that allow constant motor speed while varying the load speed. The Variable Frequency Drive Evaluation Protocol presented here addresses evaluation issues for variable-frequency drives (VFDs) installed on commercial and industrial motor-driven centrifugal fans and pumps for which torque varies with speed. Constant torque load applications, such as those for positive displacement pumps, are not covered by this protocol.

A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis.

PubMed

Phan, Trang; Huynh, Phuong; Truong, Tuom; Nguyen, Hoang

2017-01-01

Bacillus subtilis (B. subtilis) is a potential and attractive host for the production of recombinant proteins. Different expression systems for B. subtilis have been developed recently, and various target proteins have been recombinantly synthesized and purified using this host. In this chapter, we introduce a generic protocol to express a recombinant protein in B. subtilis. It includes protocols for (1) using our typical expression vector (plasmid pHT254) to introduce a target gene, (2) transformation of the target vector into B. subtilis, and (3) evaluation of the actual expression of a recombinant protein.

New York University's Stern School of Business: A Centennial Retrospective.

ERIC Educational Resources Information Center

Gitlow, Abraham L.

This volume traces the history of the first hundred years of the Stern School of Business at New York University. Chapter 1 describes the school's original mission and founding. Chapter 2 describes how the school changed and developed as it responded to trends from 1900 to 1950. Chapter 3 explores the school's dramatic decline between 1955 and…

Somatic Cell Nuclear Transfer in the Mouse

NASA Astrophysics Data System (ADS)

Kishigami, Satoshi; Wakayama, Teruhiko

Somatic cell nuclear transfer (SCNT) has become a unique and powerful tool for epigenetic reprogramming research and gene manipulation in animals since “Dolly,” the first animal cloned from an adult cell was reported in 1997. Although the success rates of somatic cloning have been inefficient and the mechanism of reprogramming is still largely unknown, this technique has been proven to work in more than 10 mammalian species. Among them, the mouse provides the best model for both basic and applied research of somatic cloning because of its abounding genetic resources, rapid sexual maturity and propagation, minimal requirements for housing, etc. This chapter describes a basic protocol for mouse cloning using cumulus cells, the most popular cell type for NT, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. In particular, we focus on a new, more efficient mouse cloning protocol using trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, which increases both in vitro and in vivo developmental rates from twofold to fivefold. This new method including TSA will be helpful to establish mouse cloning in many laboratories.

Magnetic-Activated Cell Sorting for the Fast and Efficient Separation of Human and Rodent Schwann Cells from Mixed Cell Populations.

PubMed

Ravelo, Kristine M; Andersen, Natalia D; Monje, Paula V

2018-01-01

To date, magnetic-activated cell sorting (MACS) remains a powerful method to isolate distinct cell populations based on differential cell surface labeling. Optimized direct and indirect MACS protocols for cell immunolabeling are presented here as methods to divest Schwann cell (SC) cultures of contaminating cells (specifically, fibroblast cells) and isolate SC populations at different stages of differentiation. This chapter describes (1) the preparation of single-cell suspensions from established human and rat SC cultures, (2) the design and application of cell selection strategies using SC-specific (p75 NGFR , O4, and O1) and fibroblast-specific (Thy-1) markers, and (3) the characterization of both the pre- and post-sorting cell populations. A simple protocol for the growth of hybridoma cell cultures as a source of monoclonal antibodies for cell surface immunolabeling of SCs and fibroblasts is provided as a cost-effective alternative for commercially available products. These steps allow for the timely and efficient recovery of purified SC populations without compromising the viability and biological activity of the cells.

In Vitro Cryopreservation of Date Palm Caulogenic Meristems.

PubMed

Fki, Lotfi; Chkir, Olfa; Kriaa, Walid; Nasri, Ameni; Baklouti, Emna; Masmoudi, Raja B; Rival, Alain; Drira, Noureddine; Panis, Bart

2017-01-01

Cryopreservation is the technology of choice not only for plant genetic resource preservation but also for virus eradication and for the efficient management of large-scale micropropagation. In this chapter, we describe three cryopreservation protocols (standard vitrification, droplet vitrification, and encapsulation vitrification) for date palm highly proliferating meristems that are initiated from vitro-cultures using plant growth regulator-free MS medium. The positive impact of sucrose preculture and cold hardening treatments on survival rates is significant. Regeneration rates obtained with standard vitrification, encapsulation-vitrification, and droplet-vitrification protocols can reach 30, 40, and 70%, respectively. All regenerated plants from non-cryopreserved or cryopreserved explants don't show morphological variation by maintaining genetic integrity without adverse effect of cryogenic treatment. Cryopreservation of date palm vitro-cultures enables commercial tissue culture laboratories to move to large-scale propagation from cryopreserved cell lines producing true-to-type plants after clonal field-testing trials. When comparing the cost of cryostorage and in-field conservation of date palm cultivars, tissue cryopreservation is the most cost-effective. Moreover, many of the risks linked to field conservation like erosion due to climatic, edaphic, and phytopathologic constraints are circumvented.

Molecular Typing and Differentiation

EPA Science Inventory

In this chapter, general background and bench protocols are provided for a number of molecular typing techniques in common use today. Methods for the molecular typing and differentiation of microorganisms began to be widely adopted following the development of the polymerase chai...

Recent advances in high-pressure freezing: equipment- and specimen-loading methods.

PubMed

McDonald, Kent L; Morphew, Mary; Verkade, Paul; Müller-Reichert, Thomas

2007-01-01

This chapter is an update of material first published by McDonald in the first volume of this book. Here, we discuss the improvements in the technology and the methodology of high-pressure freezing (HPF) since that article was published. First, we cover the latest innovation in HPF, the Leica EM PACT2. This machine differs significantly from the BAL-TEC HPM 010 high-pressure freezer, which was the main subject of the former chapter. The EM PACT2 is a smaller, portable machine and has an optional attachment, the Rapid Transfer System (RTS). This RTS permits easy and reproducible loading of the sample and allows one to do correlative light and electron microscopy with high time resolution. We also place more emphasis in this article on the details of specimen loading for HPF, which is considered the most critical phase of the whole process. Detailed procedures are described for how to high-pressure freeze cells in suspension, cells attached to substrates, tissue samples, or whole organisms smaller than 300 microm, and tissues or organisms greater than 300 microm in size. We finish the article with a brief discussion of freeze substitution and recommend some sample protocols for this procedure.

Presentation of nursing diagnosis content in fundamentals of nursing textbooks.

PubMed

Mahon, S M; Spies, M A; Aukamp, V; Barrett, J T; Figgins, M J; Meyer, G A; Young, V K

1997-01-01

The technique and rationale for the use of nursing diagnosis generally are introduced early in the undergraduate curriculum. The three purposes of this descriptive study were to describe the general characteristics and presentation of content on nursing diagnosis in fundamentals of nursing textbooks; describe how the content from the theoretical chapter(s) in nursing diagnosis is carried through in the clinical chapters; and describe how content on diagnostic errors is presented. Although most of the textbooks presented content on nursing diagnosis in a similar fashion, the clinical chapters of the books did not follow the same pattern. Content on diagnostic errors was inconsistent. Educators may find this an effective methodology for reviewing textbooks.

22 CFR Appendix B to Chapter Xiv - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2012 CFR

2012-04-01

...-Management Relations Statute B Appendix B to Chapter XIV Foreign Relations FOREIGN SERVICE LABOR RELATIONS... THE FOREIGN SERVICE IMPASSE DISPUTES PANEL Ch. XIV, App. B Appendix B to Chapter XIV—Memorandum... under this chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other...

22 CFR Appendix B to Chapter Xiv - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2011 CFR

2011-04-01

...-Management Relations Statute B Appendix B to Chapter XIV Foreign Relations FOREIGN SERVICE LABOR RELATIONS... THE FOREIGN SERVICE IMPASSE DISPUTES PANEL Ch. XIV, App. B Appendix B to Chapter XIV—Memorandum... under this chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other...

22 CFR Appendix B to Chapter Xiv - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2013 CFR

2013-04-01

...-Management Relations Statute B Appendix B to Chapter XIV Foreign Relations FOREIGN SERVICE LABOR RELATIONS... THE FOREIGN SERVICE IMPASSE DISPUTES PANEL Ch. XIV, App. B Appendix B to Chapter XIV—Memorandum... under this chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other...

22 CFR Appendix B to Chapter Xiv - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2010 CFR

2010-04-01

...-Management Relations Statute B Appendix B to Chapter XIV Foreign Relations FOREIGN SERVICE LABOR RELATIONS... THE FOREIGN SERVICE IMPASSE DISPUTES PANEL Ch. XIV, App. B Appendix B to Chapter XIV—Memorandum... under this chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other...

The life stories of adults with amnesia: Insights into the contribution of the medial temporal lobes to the organization of autobiographical memory.

PubMed

Grilli, Matthew D; Wank, Aubrey A; Verfaellie, Mieke

2018-02-01

Autobiographical memories are not stored in isolation but rather are organized into life chapters, higher-order knowledge structures that represent major themes conveying the arc of one's life. Neuropsychological studies have revealed that both episodic memory and some aspects of personal semantic memory are impaired in adults with medial temporal lobe (MTL) damage. However, whether such impairment compromises the retrieval and formation of life chapters is unknown. Therefore, we had 10 adults with MTL amnesia and 20 control participants narrate their life stories, and we extracted life chapters from these narratives using a novel scoring protocol. For the retrograde and anterograde time period separately, we evaluated the number of life chapters and assessed their quality, as indexed by measures of complexity and richness. Additionally, to investigate the idea that formation of life chapters occurs on a protracted time scale, we separated the amnesic participants into an early-life and a later-life onset subgroup. Results revealed that early-onset, but not later-onset, amnesic participants generated fewer retrograde life chapters than controls. The higher-order temporal relation among retrograde chapters, but not their thematic relation or the richness of individual life chapters, was impaired in both amnesic subgroups. The amnesic participants also generated fewer anterograde life chapters than controls, and the richness of their anterograde chapters was reduced in terms of content, but not self-reflection. Findings suggest that the organization of autobiographical content into life chapters is a protracted process that depends on the MTL, as does retrieval of higher order temporal relations among life chapters. Copyright © 2017 Elsevier Ltd. All rights reserved.

Status of volcanism studies for the Yucca Mountain Site Characterization Project

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

Crowe, B.; Perry, F.; Murrell, M.

1995-02-01

Chapter 1 introduces the volcanism issue for the Yucca Mountain site and provides the reader with an overview of the organization, content, and significant conclusions of this report. The risk of future basaltic volcanism is the primary topic of concern including both events that intersect a potential repository and events that occur near or within the waste isolation system of a repository. Chapter 2 describes the volcanic history of the Yucca Mountain region (YMR) and emphasizes the Pliocene and Quaternary volcanic record, the interval of primary concern for volcanic risk assessment. The Lathrop Wells volcanic center is described in detailmore » because it is the youngest basalt center in the YMR. Chapter 3 describes the tectonic setting of the YMR and presents and assesses the significance of multiple alternative tectonic models. Geophysical data are described for the YMR and are used as an aid to understand the distribution of basaltic volcanic centers. Chapter 4 discusses the petrologic and geochemical features of basaltic volcanism in the YMR, the southern Great Basin and the Basin and Range province. The long time of activity and characteristic small volume of the Postcaldera basalt of the YMR result in one of the lowest eruptive rates in a volcanic field in the southwest United States. Chapter 5 summarizes current concepts of the segregation, ascent, and eruption of basalt magma. Chapter 6 summarizes the history of volcanism studies (1979 through early 1994), including work for the Yucca Mountain Site Characterization Project and overview studies by the state of Nevada and the Nuclear Regulatory Commission. Chapter 7 summarizes probabilistic volcanic hazard assessment using a three-part conditional probability model. Chapter 8 describes remaining volcanism work judged to be needed to complete characterization studies for the YMR. Chapter 9 summarizes the conclusions of this volcanism status report.« less

HABITAT ASSESSMENT METHODS

EPA Science Inventory

This chapter summarizes and evaluated the habitat assessment protocols of five agencies, USEPA/EMAP/SW, USGS/NAWQA, USEPA/RBP, Ohio EPA, and MDNR/MBSS. It begins with a description of the origin of the habitat indices most widely used by these agencies. Then the habitat assessmen...

Protocols for In Vitro Mass Multiplication and Analysis of Medicinally Important Phenolics of a Salep Orchid, Satyrium nepalense D.Don ("Salam Mishri").

PubMed

Babbar, Shashi B; Singh, Deepak K

2016-01-01

Satyrium nepalense is a rare and threatened medicinal orchid, populations of which in its native habitats are dwindling because of indiscriminate collections and habitat destruction, thus necessitating the development of methods for its in situ and ex situ conservation. Because of non-endospermous nature of the seeds and the immature embryos at seed dispersal stage, orchids cannot be seed-propagated as other plants. Micropropagation, using plant tissue culture techniques, offers an effective method for the multiplication of orchids. In this chapter, a five-step efficient reproducible protocol for large-scale in vitro multiplication of Satyrium nepalense is described. The first step involves asymbiotic germination of seeds isolated from immature green pods and cultured on Mitra's medium (M) gelled with 0.8 % agar and supplemented with 2 % sucrose and 1 % peptone (hereafter referred to as basal medium, BM). On this medium, seeds start germinating after a week of culture. Protocorms developed from the seeds are sub-cultured on BM fortified with 4 μM kinetin (Kn) after 8 weeks, for shoot differentiation and multiplication. The shoots developed on Kn-supplemented medium are transferred to BM alone for their elongation for the same period. The elongated shoots are transferred to the rooting medium, comprising BM supplemented with 0.5 or 1.0 μM indole-3-butyric acid, for further 8 weeks. The regenerated plantlets are transferred to a potting mix of sand and vermiculite (1:1) for acclimatization. The tubers and leaves excised from both in vitro-developed plants and those from their native habitats are analyzed and compared for the contents and concentration of medicinally important phenolics using high-performance liquid chromatography (HPLC), details of which are provided in this chapter.

Gossip-Based Broadcast

NASA Astrophysics Data System (ADS)

Leitão, João; Pereira, José; Rodrigues, Luís

Gossip, or epidemic, protocols have emerged as a powerful strategy to implement highly scalable and resilient reliable broadcast primitives on large scale peer-to-peer networks. Epidemic protocols are scalable because they distribute the load among all nodes in the system and resilient because they have an intrinsic level of redundancy that masks node and network failures. This chapter provides an introduction to gossip-based broadcast on large-scale unstructured peer-to-peer overlay networks: it surveys the main results in the field, discusses techniques to build and maintain the overlays that support efficient dissemination strategies, and provides an in-depth discussion and experimental evaluation of two concrete protocols, named HyParView and Plumtree.

Chapter 4- Fertility preservation in women with breast cancer

PubMed Central

Rodriguez-Wallberg, Kenny A.; Oktay, Kutluk

2010-01-01

Fertility preservation is an important issue for young women diagnosed with breast cancer. The most well-established options for fertility preservation in cancer patients, embryo and oocyte cryopreservation, have not been traditionally offered to breast cancer patients as estradiol rise during standard stimulation protocols may not be safe for those patients. Potentially safer stimulation protocols using tamoxifen and aromatase inhibitors induce lower levels of estradiol while similar results in terms of number of oocyte and embryo obtained to standard protocols. Cryopreservation of immature oocytes and ovarian cortical tissue, both still experimental methods, are also fertility preservation options for breast cancer patients. PMID:21048442

Analytical chemistry at the interface between materials science and biology

NASA Astrophysics Data System (ADS)

O'Brien, Janese Christine

This work describes several research efforts that lie at the new interfaces between analytical chemistry and other disciplines, namely materials science and biology. In the materials science realm, the search for new materials that may have useful or unique chromatographic properties motivated the synthesis and characterization of electrically conductive sol-gels. In the biology realm, the search for new surface fabrication schemes that would permit or even improve the detection of specific biological reactions motivated the design of miniaturized biological arrays. Collectively, this work represents some of analytical chemistry's newest forays into these disciplines. This dissertation is divided into six chapters. Chapter 1 is an introductory chapter that provides background information pertinent to several key aspects of the work contained in this dissertation. Chapter 2 describes the synthesis and characterization of electrically conductive sol-gels derived from the acid-catalyzed hydrolysis of a vanadium alkoxide. Specifically, this chapter describes our attempts to increase the conductivity of vanadium sol-gels by optimizing the acidic and drying conditions used during synthesis. Chapter 3 reports the construction of novel antigenic immunosensing platforms of increased epitope density using Fab'-SH antibody fragments on gold. Here, X-ray photoelectron spectroscopy (XPS), thin-layer cell (TLC) and confocal fluorescence spectroscopies, and scanning force microscopy (SFM) are employed to characterize the fragment-substrate interaction, to quantify epitope density, and to demonstrate fragment viability and specificity. Chapter 4 presents a novel method for creating and interrogating double-stranded DNA (dsDNA) microarrays suitable for screening protein:dsDNA interactions. Using the restriction enzyme ECoR1, we demonstrate the ability of the atomic force microscope (AFM) to detect changes in topography that result from the enzymatic cleavage of dsDNA microarrays containing the correct recognition sequence. Chapter 5 explores more fully the microarray fabrication process described in Chapter 4. Specifically, experiments characterizing the effect of deposition conditions on oligonucleotide topography and as well as those that describe array density optimization are presented. Chapter 6 presents general conclusions from the work recorded in this dissertation and speculates on its extension.

22 CFR Appendix B to Chapter Xiv - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2014 CFR

2014-04-01

... Service Labor-Management Relations Statute B Appendix B to Chapter XIV Foreign Relations FOREIGN SERVICE... AUTHORITY; AND THE FOREIGN SERVICE IMPASSE DISPUTES PANEL Ch. XIV, App. B Appendix B to Chapter XIV... under this chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other...

Grapevine (Vitis vinifera L.).

PubMed

Torregrosa, Laurent; Vialet, Sandrine; Adivèze, Angélique; Iocco-Corena, Pat; Thomas, Mark R

2015-01-01

Grapevine (Vitis) is considered to be one of the major fruit crops in the world based on hectares cultivated and economic value. Grapes are used not only for wine but also for fresh fruit, dried fruit, and juice production. Wine is by far the major product of grapes, and the focus of this chapter is on wine grape cultivars. Grapevine cultivars of Vitis vinifera L. have a reputation for producing premium quality wines. These premium quality wines are produced from a small number of cultivars that enjoy a high level of consumer acceptance and are firmly entrenched in the market place because of varietal name branding and the association of certain wine styles and regions with specific cultivars. In light of this situation, grapevine improvement by a transgenic approach is attractive when compared to a classical breeding approach. The transfer of individual traits as single genes with a minimum disruption to the original genome would leave the traditional characteristics of the cultivar intact. However, a reliable transformation system is required for a successful transgenic approach to grapevine improvement. There are three criteria for achieving an efficient Agrobacterium-mediated transformation system: (1) the production of highly regenerative transformable tissue, (2) optimal cocultivation conditions for both grapevine tissue and Agrobacterium, and (3) an efficient selection regime for transgenic plant regeneration. In this chapter, we describe a grapevine transformation system that meets these criteria. We also describe a protocol for the production of transformed roots suitable for functional gene studies and for the production of semi-transgenic grafted plants.

Training for Development 1970. The United States Participant Training Program.

ERIC Educational Resources Information Center

Agency for International Development (Dept. of State), Washington, DC. Office of International Training.

The report describes the program of training for foreign students conducted by the Office of International Training of the Agency for International Development (AID). The report's first chapter describes the history, nature, and importance of participant training and diagrams the administrative structure of the program. Chapter 2 describes the…

Characterizing millisecond intermediates in hemoproteins using rapid-freeze-quench resonance Raman spectroscopy.

PubMed

Matsumura, Hirotoshi; Moënne-Loccoz, Pierre

2014-01-01

The combination of rapid freeze quenching (RFQ) with resonance Raman (RR) spectroscopy represents a unique tool with which to investigate the nature of short-lived intermediates formed during the enzymatic reactions of metalloproteins. Commercially available equipment allows trapping of intermediates within a millisecond to second time scale for low-temperature RR analysis resulting in the direct detection of metal-ligand vibrations and porphyrin skeletal vibrations in hemoproteins. This chapter briefly discusses RFQ-RR studies carried out previously in our laboratory and presents, as a practical example, protocols for the preparation of RFQ samples of the reaction of metmyoglobin with nitric oxide (NO) under anaerobic conditions. Also described are important controls and practical procedures for the analysis of these samples by low-temperature RR spectroscopy.

Simplified methods for the construction of RNA and DNA virus infectious clones.

PubMed

Nagata, Tatsuya; Inoue-Nagata, Alice Kazuko

2015-01-01

Infectious virus clones are one of the most powerful tools in plant pathology, molecular biology, and biotechnology. The construction of infectious clones of RNA and DNA viruses, however, usually requires laborious cloning and subcloning steps. In addition, instability of the RNA virus genome is frequently reported after its introduction into the vector and transference to Escherichia coli. These difficulties hamper the cloning procedures, making it tedious and cumbersome. This chapter describes two protocols for a simple construction of infectious viruses, an RNA virus, the tobamovirus Pepper mild mottle virus, and a DNA virus, a bipartite begomovirus. For this purpose, the strategy of overlap-extension PCR was used for the construction of infectious tobamovirus clone and of rolling circle amplification (RCA) for the construction of a dimeric form of the begomovirus clone.

Analysis of nuclear export using photoactivatable GFP fusion proteins and interspecies heterokaryons.

PubMed

Nakrieko, Kerry-Ann; Ivanova, Iordanka A; Dagnino, Lina

2010-01-01

In this chapter, we review protocols for the analysis of nucleocytoplasmic shuttling of transcription factors and nuclear proteins, using two different approaches. The first involves the use of photoactivatable forms of the protein of interest by fusion to photoactivatable green fluorescent protein to follow its movement out of the nucleus by live-cell confocal microscopy. This methodology allows for the kinetic characterization of protein movements as well as measurement of steady-state levels. In a second procedure to assess the ability of a nuclear protein to move into and out of the nucleus, we describe the use of interspecies heterokaryon assays, which provide a measurement of steady-state distribution. These technologies are directly applicable to the analysis of nucleocytoplasmic movements not only of transcription factors, but also other nuclear proteins.

Combining Phage and Yeast Cell Surface Antibody Display to Identify Novel Cell Type-Selective Internalizing Human Monoclonal Antibodies.

PubMed

Bidlingmaier, Scott; Su, Yang; Liu, Bin

2015-01-01

Using phage antibody display, large libraries can be generated and screened to identify monoclonal antibodies with affinity for target antigens. However, while library size and diversity is an advantage of the phage display method, there is limited ability to quantitatively enrich for specific binding properties such as affinity. One way of overcoming this limitation is to combine the scale of phage display selections with the flexibility and quantitativeness of FACS-based yeast surface display selections. In this chapter we describe protocols for generating yeast surface antibody display libraries using phage antibody display selection outputs as starting material and FACS-based enrichment of target antigen-binding clones from these libraries. These methods should be widely applicable for the identification of monoclonal antibodies with specific binding properties.

Single Cell Gene Expression Profiling of Skeletal Muscle-Derived Cells.

PubMed

Gatto, Sole; Puri, Pier Lorenzo; Malecova, Barbora

2017-01-01

Single cell gene expression profiling is a fundamental tool for studying the heterogeneity of a cell population by addressing the phenotypic and functional characteristics of each cell. Technological advances that have coupled microfluidic technologies with high-throughput quantitative RT-PCR analyses have enabled detailed analyses of single cells in various biological contexts. In this chapter, we describe the procedure for isolating the skeletal muscle interstitial cells termed Fibro-Adipogenic Progenitors (FAPs ) and their gene expression profiling at the single cell level. Moreover, we accompany our bench protocol with bioinformatics analysis designed to process raw data as well as to visualize single cell gene expression data. Single cell gene expression profiling is therefore a useful tool in the investigation of FAPs heterogeneity and their contribution to muscle homeostasis.

Studies of proteoglycan involvement in CPP-mediated delivery.

PubMed

Wittrup, Anders; Zhang, Si-He; Belting, Mattias

2011-01-01

Cell-penetrating peptides (CPPs) are widely used to deliver macromolecular cargoes to intracellular sites of action. Many CPPs have been demonstrated to rely on cell surface heparan sulfate proteoglycans (HSPGs) for efficient cellular entry and delivery. In this chapter, we describe methods for the study of PG involvement in CPP uptake. We provide descriptions of how to determine whether uptake of a CPP of interest is dependent on PGs. We also provide detailed protocols for the purification of PGs by anion-exchange chromatography as well as the characterization of the HSPG core protein composition of a cell line of interest. Finally, we present methods for modulating the expression level of specific HSPG core proteins as a means to determine the core protein specificity in the uptake of a particular CPP.

Image Analysis of DNA Fiber and Nucleus in Plants.

PubMed

Ohmido, Nobuko; Wako, Toshiyuki; Kato, Seiji; Fukui, Kiichi

2016-01-01

Advances in cytology have led to the application of a wide range of visualization methods in plant genome studies. Image analysis methods are indispensable tools where morphology, density, and color play important roles in the biological systems. Visualization and image analysis methods are useful techniques in the analyses of the detailed structure and function of extended DNA fibers (EDFs) and interphase nuclei. The EDF is the highest in the spatial resolving power to reveal genome structure and it can be used for physical mapping, especially for closely located genes and tandemly repeated sequences. One the other hand, analyzing nuclear DNA and proteins would reveal nuclear structure and functions. In this chapter, we describe the image analysis protocol for quantitatively analyzing different types of plant genome, EDFs and interphase nuclei.

Herpes simplex virus mutant generation and dual-detection methods for gaining insight into latent/lytic cycles in vivo.

PubMed

Sawtell, Nancy M; Thompson, Richard L

2014-01-01

Two important components to a useful strategy to examine viral gene regulation in vivo are (1) a highly efficient protocol to generate viral mutants that limits undesired mutation and retains full replication competency in vivo and (2) an efficient system to detect and quantify viral promoter activity in rare cells in vivo. Our strategy and protocols for generating, characterizing, and employing HSV viral promoter/reporter mutants in vivo are provided in this two-part chapter.

Production of cloned mice from somatic cells, ES cells, and frozen bodies.

PubMed

Wakayama, Sayaka; Mizutani, Eiji; Wakayama, Teruhiko

2010-01-01

Somatic cell nuclear transfer (SCNT) has become a unique and powerful tool for epigenetic reprogramming research and gene manipulation in animals. Although the success rates of somatic cloning have been inefficient and the mechanism of reprogramming is still largely unknown, therefore, the nuclear transfer (NT) method has been thought of as a "black box approach" and inadequate to determine the detail of how genomic reprogramming occurs. However, only the NT approach can reveal dynamic and global modifications in the epigenome without using genetic modification, as well as can create live animals. At present, this is the only technique available for the preservation and propagation of valuable genetic resources from mutant mice that are infertile or too old, or recovered from carcasses, without the use of germ cells. This chapter describes a basic protocol for mouse cloning and embryonic stem (ES) cell establishment from cloned embryo using a piezo-actuated micromanipulator. This technique will greatly help not only in mouse cloning but also in other forms of micromanipulation such as intracytoplasmic sperm injection (ICSI) into oocytes or ES cell injection into blastocysts. In addition, we describe a new, more efficient mouse cloning protocol using histone deacetylase inhibitor (HDACi), which increases the success rates of cloned mice or establish rate of ES cells to fivefold. Copyright (c) 2010 Elsevier Inc. All rights reserved.

Transit-oriented development compendium.

DOT National Transportation Integrated Search

2005-06-01

The compendium is organized into eight chapters listed below. Each chapter describes relevant issues, the state of the practice for TOD, and includes information from guidebooks and interviews. The eight chapters include: 1) general principles of TOD...

Geology and undiscovered resource assessment of the potash-bearing Central Asia Salt Basin, Turkmenistan, Uzbekistan, Tajikistan, and Afghanistan: Chapter AA in Global mineral resource assessment

USGS Publications Warehouse

Wynn, Jeff; Orris, Greta J.; Dunlap, Pamela; Cocker, Mark D.; Bliss, James D.

2016-03-23

Chapter 1 of this report provides an overview of the history of the CASB and summarizes evaporite potash deposition, halokinesis, and dissolution processes that have affected the current distribution of potash-bearing salt in the CASB. Chapter 2 describes the Gissar tract, an uplifted region that contains a mix of stratabound and halokinetic potash deposits and all of the discovered and exploited potash deposits of the CASB. Chapter 3 describes the Amu Darya tract, where evaporite deposits remain flat-lying and undeformed since their original deposition. Chapter 4 describes the highly deformed and compressed Afghan-Tajik tract and what is known of the deeply-buried Jurassic salt. Chapter 5 describes the spatial databases included with this report, which contain a collection of CASB potash information. Appendixes A and B summarize descriptive models for stratabound and halokinetic potash-bearing salt deposits, respectively. Appendix C summarizes the AGE method used to evaluate the Gissar and Amu Darya tracts. Appendixes D and E contain grade and thickness data for the Gissar and Amu Darya tracts. Appendix F provides the SYSTAT script used to estimate undiscovered K2 O in a CASB tract. Appendix G provides a potash glossary, and appendix H provides biographies of assessment participants.

Histological and morphometric analyses for rat carotid balloon injury model.

PubMed

Tulis, David A

2007-01-01

Experiments aimed at analyzing the response of blood vessels to mechanical injury and ensuing remodeling responses often employ the highly characterized carotid artery balloon injury model in laboratory rats. This approach utilizes luminal insertion of a balloon embolectomy catheter into the common carotid artery with inflation and withdrawal resulting in an injury characterized by vascular endothelial cell (EC) denudation and medial wall distension. The adaptive response to this injury is typified by robust vascular smooth muscle cell (SMC) replication and migration, SMC apoptosis and necrosis, enhanced synthesis and deposition of extracellular matrix (ECM) components, partial vascular EC regeneration from the border zones, luminal narrowing, and establishment of a neointima in time-dependent fashion. Evaluation of these adaptive responses to blood vessel injury can include acute and longer term qualitative and quantitative measures including expression analyses, activity assays, immunostaining for a plethora of factors and signals, and morphometry of neointima formation and gross mural remodeling. This chapter presents a logical continuation of Chapter 1 that offers details for performing the rat carotid artery balloon injury model in a standard laboratory setting by providing commonly used protocols for performing histological and morphometric analyses in such studies. Moreover, procedures, caveats, and considerations included in this chapter are highly relevant for alternative animal vascular physiology/pathophysiology studies and in particular those related to mechanisms of vascular injury and repair. Included in this chapter are specifics for in situ perfusion-fixation, tissue harvesting and processing for both snap-frozen and paraffin-embedded protocols, specimen embedding and sectioning, slide preparation, several standard histological staining steps, and routine morphological assessment.

Peptide-membrane Interactions by Spin-labeling EPR

PubMed Central

Smirnova, Tatyana I.; Smirnov, Alex I.

2016-01-01

Site-directed spin labeling (SDSL) in combination with Electron Paramagnetic Resonance (EPR) spectroscopy is a well-established method that has recently grown in popularity as an experimental technique, with multiple applications in protein and peptide science. The growth is driven by development of labeling strategies, as well as by considerable technical advances in the field, that are paralleled by an increased availability of EPR instrumentation. While the method requires an introduction of a paramagnetic probe at a well-defined position in a peptide sequence, it has been shown to be minimally destructive to the peptide structure and energetics of the peptide-membrane interactions. In this chapter, we describe basic approaches for using SDSL EPR spectroscopy to study interactions between small peptides and biological membranes or membrane mimetic systems. We focus on experimental approaches to quantify peptide-membrane binding, topology of bound peptides, and characterize peptide aggregation. Sample preparation protocols including spin-labeling methods and preparation of membrane mimetic systems are also described. PMID:26477253

Measuring suspended sediment: Chapter 10

USGS Publications Warehouse

Gray, J.R.; Landers, M.N.

2013-01-01

Suspended sediment in streams and rivers can be measured using traditional instruments and techniques and (or) surrogate technologies. The former, as described herein, consists primarily of both manually deployed isokinetic samplers and their deployment protocols developed by the Federal Interagency Sedimentation Project. They are used on all continents other than Antarctica. The reliability of the typically spatially rich but temporally sparse data produced by traditional means is supported by a broad base of scientific literature since 1940. However, the suspended sediment surrogate technologies described herein – based on hydroacoustic, nephelometric, laser, and pressure difference principles – tend to produce temporally rich but in some cases spatially sparse datasets. The value of temporally rich data in the accuracy of continuous sediment-discharge records is hard to overstate, in part because such data can often overcome the shortcomings of poor spatial coverage. Coupled with calibration data produced by traditional means, surrogate technologies show considerable promise toward providing the fluvial sediment data needed to increase and bring more consistency to sediment-discharge measurements worldwide.

The application of spaceborne GPS to atmospheric limb sounding and global change monitoring

NASA Technical Reports Server (NTRS)

Melbourne, W. G.; Davis, E. S.; Duncan, C. B.; Hajj, G. A.; Hardy, K. R.; Kursinski, E. R.; Meehan, T. K.; Young, L. E.; Yunck, T. P.

1994-01-01

This monograph is intended for readers with minimal background in radio science who seek a relatively comprehensive treatment of the mission and technical aspects of an Earth-orbiting radio occultation satellite. Part 1 (chapters 1-6) describes mission concepts and programmatic information; Part 2 (chapters 7-12) deals with the theoretical aspects of analyzing and interpreting radio occultation measurements. In this mission concept the navigation signals from a Global Positioning System (GPS) satellite that is being occulted by the Earth's limb are observed by a GPS flight receiver on board a low Earth orbiter (LEO) satellite. This technique can be used to recover profiles of the Earth's atmospheric refractivity, pressure, and temperature using small, dedicated, and relatively low-cost space systems. Chapter 2 summarizes the basic space system concepts of the limb-sounding technique and describes a low-cost strawman demonstration mission. Chapter 3 discusses some of the scientific benefits of using radio occultation on a suite of small satellites. Chapter 4 provides a more detailed discussion of several system elements in a radio occultation mission, including the launch system for small payloads, the LEO microsat, the GPS constellation, the GPS flight receiver payload, the mission operations ground control and data receiving system, the ground-based GPS global tracking network for precision orbit determination, and the central data processing and archive system. Chapter 5 addresses the various technology readiness questions that invariably arise. Chapter 6 discusses the overall costs of a demonstration mission such as GPS/MET (meteorological) proposed by the University Navstar Consortium (UNAVCO). Chapter 7 describes a geometrical optics approach to coplanar atmospheric occultation. Chapter 8 addresses major questions regarding accuracy of the occultation techniques. Chapter 9 describes some simulations that have been performed to evaluate the sensitivity of the recovered profiles of atmospheric parameters to different error sources, such as departure from spherical symmetry, water vapor, etc. Chapter 10 discusses horizontal and vertical resolution associated with limb sounders in general. Chapter 11 treats selected Fresnel diffraction techniques that can be used in radio occultation measurements to sharpen resolution. Chapter 12 provides brief discussions on selected special topics, such as strategies for handling interference and multipath processes that may arise for rays traveling in the lower troposphere.

Radiation Protection Studies of International Space Station Extravehicular Activity Space Suits

NASA Technical Reports Server (NTRS)

Cucinotta, Francis A. (Editor); Shavers, Mark R. (Editor); Saganti, Premkumar B. (Editor); Miller, Jack (Editor)

2003-01-01

This publication describes recent investigations that evaluate radiation shielding characteristics of NASA's and the Russian Space Agency's space suits. The introduction describes the suits and presents goals of several experiments performed with them. The first chapter provides background information about the dynamic radiation environment experienced at ISS and summarized radiation health and protection requirements for activities in low Earth orbit. Supporting studies report the development and application of a computer model of the EMU space suit and the difficulty of shielding EVA crewmembers from high-energy reentrant electrons, a previously unevaluated component of the space radiation environment. Chapters 2 through 6 describe experiments that evaluate the space suits' radiation shielding characteristics. Chapter 7 describes a study of the potential radiological health impact on EVA crewmembers of two virtually unexamined environmental sources of high-energy electrons-reentrant trapped electrons and atmospheric albedo or "splash" electrons. The radiological consequences of those sources have not been evaluated previously and, under closer scrutiny. A detailed computational model of the shielding distribution provided by components of the NASA astronauts' EMU is being developed for exposure evaluation studies. The model is introduced in Chapters 8 and 9 and used in Chapter 10 to investigate how trapped particle anisotropy impacts female organ doses during EVA. Chapter 11 presents a review of issues related to estimating skin cancer risk form space radiation. The final chapter contains conclusions about the protective qualities of the suit brought to light form these studies, as well as recommendations for future operational radiation protection.

Development of conductometric biosensors based on alkaline phosphatases for the water quality control

NASA Astrophysics Data System (ADS)

Berezhetskyy, A.

2008-09-01

Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devoted to creation and optimization of conductometric biosensor based on alkaline phosphatase active microalgae and sol gel technology, the last chapter described application of the proposed algal biosensor for measurements of heavy metal ions toxicity of waste water, general conclusions stating the progresses achieved in the field of environmental monitoring

High-pressure freezing and freeze substitution of Arabidopsis for electron microscopy.

PubMed

Austin, Jotham R

2014-01-01

The objectives of electron microscopy ultrastructural studies are to examine cellular architecture and relate the cell's structural machinery to dynamic functional roles. This aspiration is difficult to achieve if specimens have not been adequately preserved in a "living state"; hence specimen preparation is of the utmost importance for the success of any electron micrographic study. High-pressure freezing (HPF)/freeze substitution (FS) has long been recognized as the primer technique for the preservation of ultrastructure in biological samples. In most cases a basic HPF/freeze substitution protocol is sufficient to obtain superior ultrastructural preservation and structural contrast, which allows one to use more advanced microscopy techniques such as 3D electron tomography. However, for plant tissues, which have a thick cell wall, large water-filled vacuoles, and air spaces (all of which are detrimental to cryopreservation), these basic HPF/FS protocols often yield undesirable results. In particular, ice crystal artifacts and the staining of membrane systems are often poorly or negatively stained, which make 3D segmentation of a tomogram difficult. To overcome these problems, various aspects of the HPF/FS protocol can be altered, including the cryo-filler(s) used, freeze substitution cocktail, and the resin infiltration process. This chapter will describe these modifications for the preparation of plant tissues for routine electron microscopic studies, immunocytochemistry, and 3D tomographic electron imaging.

The Quality Professor: Implementing TQM in the Classroom.

ERIC Educational Resources Information Center

Cornesky, Robert A.

This volume describes Total Quality Management (TQM) in the higher education classroom and guides college faculty in implementing TQM to improve their teaching. Chapter 1 introduces TQM and gives pointers on how to begin implementing it. Chapter 2 describes TQM approaches and principles including the Deming and Crosby approaches, describes the TQM…

National Standards for Family and Consumer Sciences Education.

ERIC Educational Resources Information Center

National Association of State Administrators for Family and Consumer Sciences.

The document presents a new set of standards for family and consumer sciences (FACS) education. Section 1 is a three-chapter overview. Chapter 1 addresses the rationale for change and the FACS vision and mission. Chapter 2 describes the approach to develop the national standards, FACS format, and components of the standards. Chapter 3 provides…

Historical Perspectives on School Health.

ERIC Educational Resources Information Center

Means, Richard K.

This book describes the evolution of philosophy and practice in school health education in the United States. It is divided into 12 chapters. Chapter 1 briefly traces the evolutionary development of the field. Chapter 2 gives a chronological review of significant major movements. Chapter 3 focuses on the two curriculum problems of (a) what to…

Chapter 6: Floral Transformation of Wheat

USDA-ARS?s Scientific Manuscript database

Hexaploid wheat is one of the world’s most important staple crops but genetic transformation is still challenging. We are developing a floral transformation protocol for wheat that does not require tissue culture. Several T-DNA transformants have been produced in the high quality, hard red germpla...

Chapter 17 Sterile Plate-Based Vitrification of Adherent Human Pluripotent Stem Cells and Their Derivatives Using the TWIST Method.

PubMed

Neubauer, Julia C; Stracke, Frank; Zimmermann, Heiko

2017-01-01

Due to their high biological complexity, e.g., their close cell-to-cell contacts, cryopreservation of human pluripotent stem cells with standard slow-rate protocols often is inefficient and can hardly be standardized. Vitrification that means ultrafast freezing already showed very good viability and recovery rates for this sensitive cell system, but is only applicable for low cell numbers, bears a high risk of contamination, and can hardly be implemented under GxP regulations. In this chapter, a sterile plate-based vitrification method for adherent pluripotent stem cells and their derivatives is presented based on a procedure and device for human embryonic stem cells developed by Beier et al. (Cryobiology 66:8-16, 2013). This protocol overcomes the limitations of conventional vitrification procedures resulting in the highly efficient preservation of ready-to-use adherent pluripotent stem cells with the possibility of vitrifying cells in multi-well formats for direct application in high-throughput screenings.

Detectors for the Atacama Cosmology Telescope

NASA Astrophysics Data System (ADS)

Marriage, Tobias Andrew

The Atacama Cosmology Telescope (ACT) will make measurements of the brightness temperature anisotropy in the Cosmic Microwave Background (CMB) on degree to arcminute angular scales. The ACT observing site is located 5200 m near the top of Cerro Toco in the Atacama Desert of northern Chile. This thesis presents research on the detectors which capture the image of the CMB formed at ACT's focal plane. In the first chapter, the primary brightness temperature fluctuations in the Cosmic Microwave Background are reviewed. In Chapter 2, a calculation shows how the CMB brightness is translated by ACT to an input power to the detectors. Chapter 3 describes the ACT detectors in detail and presents the response and sensitivity of the detectors to the input power computed in Chapter 2. Chapter 4 describes the detector fabrication at NASA Goddard Space Flight Center. Chapter 5 summarizes experiments which characterize the ACT detector performance.

Chapter 6: CPV Tracking and Trackers

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

Luque-Heredia, Ignacio; Magalhaes, Pedro; Muller, Matthew

2016-04-15

This chapter explains the functional requirements of a concentrator photovoltaic (CPV) sun tracker. It derives the design specifications of a CPV tracker. The chapter presents taxonomy of trackers describing the most common tracking architectures, based on the number of axes, their relative position, and the foundation and placing of tracking drives. It deals with the structural issues related to tracker design, mainly related to structural flexure and its impact on the system's acceptance angle. The chapter analyzes the auto-calibrated sun tracking control, by describing the state of the art and its development background. It explores the sun tracking accuracy measurementmore » with a practical example. The chapter discusses tracker manufacturing and tracker field works. It reviews survey of different types of tracker designs obtained from different manufacturers. Finally, the chapter deals with IEC62817, the technical standard developed for CPV sun trackers.« less

Development of Novel Polymeric Materials for Gene Therapy and pH-Sensitive Drug Delivery: Modeling, Synthesis, Characterization, and Analysis

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

Anderson, Brian Curtis

2002-01-01

The underlying theme of this thesis is the use of polymeric materials in bioapplications. Chapters 2-5 either develop a fundamental understanding of current materials used for bioapplications or establish protocols and procedures used in characterizing and synthesizing novel materials. In chapters 6 and 7 these principles and procedures are applied to the development of materials to be used for gene therapy and drug delivery. Chapter one is an introduction to the ideas that will be necessary to understand the subsequent chapters, as well as a literature review of these topics. Chapter two is a paper that has been published inmore » the ''Journal of Controlled Release'' that examines the mechanism of drug release from a polymer gel, as well as experimental design suggestions for the evaluation of water soluble drug delivery systems. Chapter three is a paper that has been published in the ''Journal of Pharmaceutical Sciences'' that discusses the effect ionic salts have on properties of the polymer systems examined in chapter two. Chapter four is a paper published in the Materials Research Society Fall 2000 Symposium Series dealing with the design and synthesis of a pH-sensitive polymeric drug delivery device. Chapter five is a paper that has been published in the journal ''Biomaterials'' proposing a novel polymer/metal composite for use as a biomaterial in hip arthroplasty surgery. Chapter six is a paper that will appear in an upcoming volume of the Journal ''Biomaterials'' dealing with the synthesis of a novel water soluble cationic polymer with possible applications in non-viral gene therapy. Chapter seven is a paper that has been submitted to ''Macromolecules'' discussing several novel block copolymers based on poly(ethylene glycol) and poly(diethylamino ethyl methacrylate) that possess both pH-sensitive and temperature sensitive properties. Chapter eight contains a summary of the research contained in chapters 2-7 and proposes future research for the gene therapy and drug delivery projects.« less

TOTAL CULTURABLE VIRUS QUANTAL ASSAY

EPA Science Inventory

This chapter describes a quantal method for assaying culturable human enteric viruses from water matrices. The assay differs from the plaque assay described in Chapter 10 (December 1987 Revision) in that it is based upon the direct microscopic viewing of cells for virus-induced ...

Potash: a global overview of evaporate-related potash resources, including spatial databases of deposits, occurrences, and permissive tracts: Chapter S in Global mineral resource assessment

USGS Publications Warehouse

Orris, Greta J.; Cocker, Mark D.; Dunlap, Pamela; Wynn, Jeff C.; Spanski, Gregory T.; Briggs, Deborah A.; Gass, Leila; Bliss, James D.; Bolm, Karen S.; Yang, Chao; Lipin, Bruce R.; Ludington, Stephen; Miller, Robert J.; Słowakiewicz, Mirosław

2014-01-01

This report describes a global, evaporite-related potash deposits and occurrences database and a potash tracts database. Chapter 1 summarizes potash resource history and use. Chapter 2 describes a global potash deposits and occurrences database, which contains more than 900 site records. Chapter 3 describes a potash tracts database, which contains 84 tracts with geology permissive for the presence of evaporite-hosted potash resources, including areas with active evaporite-related potash production, areas with known mineralization that has not been quantified or exploited, and areas with potential for undiscovered potash resources. Chapter 4 describes geographic information system (GIS) data files that include (1) potash deposits and occurrences data, (2) potash tract data, (3) reference databases for potash deposit and tract data, and (4) representative graphics of geologic features related to potash tracts and deposits. Summary descriptive models for stratabound potash-bearing salt and halokinetic potash-bearing salt are included in appendixes A and B, respectively. A glossary of salt- and potash-related terms is contained in appendix C and a list of database abbreviations is given in appendix D. Appendix E describes GIS data files, and appendix F is a guide to using the geodatabase.

Determination of saccharides and ethanol from biomass conversion using Raman spectroscopy: Effects of pretreatment and enzyme composition

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

Shih, Chien-Ju

2010-01-01

This dissertation focuses on the development of facile and rapid quantitative Raman spectroscopy measurements for the determination of conversion products in producing bioethanol from corn stover. Raman spectroscopy was chosen to determine glucose, xylose and ethanol in complex hydrolysis and fermentation matrices. Chapter 1 describes the motives and main goals of this work, and includes an introduction to biomass, commonly used pretreatment methods, hydrolysis and fermentation reactions. The principles of Raman spectroscopy, its advantages and applications related to biomass analysis are also illustrated. Chapter 2 and 3 comprise two published or submitted manuscripts, and the thesis concludes with an appendix.more » In Chapter 2, a Raman spectroscopic protocol is described to study the efficiency of enzymatic hydrolysis of cellulose by measuring the main product in hydrolysate, glucose. Two commonly utilized pretreatment methods were investigated in order to understand their effect on glucose measurements by Raman spectroscopy. Second, a similar method was set up to determine the concentration of ethanol in fermentation broth. Both of these measurements are challenged by the presence of complex matrices. In Chapter 3, a quantitative comparison of pretreatment protocols and the effect of enzyme composition are studied using systematic methods. A multipeak fitting algorithm was developed to analyze spectra of hydrolysate containing two analytes: glucose and xylose. Chapter 4 concludes with a future perspective of this research area. An appendix describes a convenient, rapid spectrophotometric method developed to measure cadmium in water. This method requires relatively low cost instrumentation and can be used in microgravity, such as space shuttles or the International Space Station. This work was performed under the supervision of Professor Marc Porter while at Iowa State University. Research related to producing biofuel from bio-renewable resources, especially bioethanol from biomass, has grown significantly in the past decade due to the high demand and rising costs of fossil fuels. More than 3 percent of the energy consumption in the U.S. is derived from renewable biomass, mostly through industrial heat and steam production by the pulp and paper industry, and electricity generation from municipal solid waste (MSW) and forest industry residues. The utilization of food-based biomass to make fuels has been widely criticized because it may increase food shortages throughout the world and raise the cost of food. Thus, nonfood-based and plentiful lignocellulosic feedstocks, such as corn stover, perennial grass, bagasse, sorghum, wheat/rice straw, herbaceous and woody crops, have great potential to be new bio-renewable sources for energy production. Given that many varieties of biomass are available, there is need for a rapid, simple, high-throughput method to screen the conversion of many plant varieties. The most suitable species for each geographic region must be determined, as well as the optimal stage of harvest, impacts of environmental conditions (temperature, soil, pH, etc.). Various genetically modified plants should be studied in order to establish the desired biomass in bioethanol production. The main screening challenge, however, is the complexity of plant cell wall structures that make reliable and sensitive analysis difficult. To date, one of the most popular methods to produce lignocellulosic ethanol is to perform enzymatic hydrolysis followed by fermentation of the hydrolysate with yeast. There are several vital needs related to the field of chemistry that have been suggested as primary research foci needed to effectively improve lignocellulosic ethanol production. These topics include overcoming the recalcitrance of cellulosic biomass, the pervasiveness of pretreatment, advanced biological processing and better feedstocks. In this thesis, a novel approach using Raman spectroscopy has been developed to address important issues related to bioethanol generation, which will aid the research aimed to solve the topics mentioned above.« less

A Skeptic among Scholars: August Fruge on University Publishing.

ERIC Educational Resources Information Center

Fruge, August

This memoir describes August Fruge's 32 years as head of the University of California Press. Early chapters describe Berkeley, where the press is located, in the 1940s, conflicts with the Printing Department, the role of the faculty-run editorial committee, and setting new directions for the press. Three chapters describe looking for titles to…

Chapter 2 Formula, 1989-90: Major Points. Publication No. 89.32.

ERIC Educational Resources Information Center

Baenen, Nancy R.

Programs implemented in 1989-90 by the Austin (Texas) Independent School District (AISD) using Chapter 2 Formula federal funds are described. Chapter 2 Formula provides federal funds to the states through the Elementary and Secondary Education Act of 1965 as amended in 1988. Chapter 2 funds can support programs that meet the educational needs of…

Kansas Chapter 1 1992-1993 Evaluation.

ERIC Educational Resources Information Center

Kansas State Board of Education, Topeka.

The 1992-93 analysis of the Kansas Chapter 1 evaluation results is described. In the 1992-93 school year, 100 percent (304) of Kansas districts participated in the Chapter 1 program. Approximately 7 percent of the student population participated in Chapter 1 programs, and 82 percent were in grades 1 through 6. A majority of these students were…

MPACT Theory Manual, Version 2.2.0

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

Downar, Thomas; Collins, Benjamin S.; Gehin, Jess C.

2016-06-09

This theory manual describes the three-dimensional (3-D) whole-core, pin-resolved transport calculation methodology employed in the MPACT code. To provide sub-pin level power distributions with sufficient accuracy, MPACT employs the method of characteristics (MOC) solutions in the framework of a 3-D coarse mesh finite difference (CMFD) formulation. MPACT provides a 3D MOC solution, but also a 2D/1D solution in which the 2D planar solution is provided by MOC and the axial coupling is resolved by one-dimensional (1-D) lower order (diffusion or P3) solutions. In Chapter 2 of the manual, the MOC methodology is described for calculating the regional angular and scalarmore » fluxes from the Boltzmann transport equation. In Chapter 3, the 2D/1D methodology is described, together with the description of the CMFD iteration process involving dynamic homogenization and solution of the multigroup CMFD linear system. A description of the MPACT depletion algorithm is given in Chapter 4, followed by a discussion of the subgroup and ESSM resonance processing methods in Chapter 5. The final Chapter 6 describes a simplified thermal hydraulics model in MPACT.« less

CHLORINE DIOXIDE CHEMISTRY, REACTIONS, AND DISINFECTION BY-PRODUCTS

EPA Science Inventory

This chapter contains two main sections-the first section describes the chemistry and reactions of chlorine dioxide, and the second describes the disinfection by-products (DBPs) of chlorine dioxide and their control. A short section on Research Needs completes this chapter. The...

Planning and setting objectives in field studies: Chapter 2

USGS Publications Warehouse

Fisher, Robert N.; Dodd, C. Kenneth

2016-01-01

This chapter enumerates the steps required in designing and planning field studies on the ecology and conservation of reptiles, as these involve a high level of uncertainty and risk. To this end, the chapter differentiates between goals (descriptions of what one intends to accomplish) and objectives (the measurable steps required to achieve the established goals). Thus, meeting a specific goal may require many objectives. It may not be possible to define some of them until certain experiments have been conducted; often evaluations of sampling protocols are needed to increase certainty in the biological results. And if sampling locations are fixed and sampling events are repeated over time, then both study-specific covariates and sampling-specific covariates should exist. Additionally, other critical design considerations for field study include obtaining permits, as well as researching ethics and biosecurity issues.

Study of AMPK-Regulated Metabolic Fluxes in Neurons Using the Seahorse XFe Analyzer.

PubMed

Marinangeli, Claudia; Kluza, Jérome; Marchetti, Philippe; Buée, Luc; Vingtdeux, Valérie

2018-01-01

AMP-activated protein kinase (AMPK) is the intracellular master energy sensor and metabolic regulator. AMPK is involved in cell energy homeostasis through the regulation of glycolytic flux and mitochondrial biogenesis. Interestingly, metabolic dysfunctions and AMPK deregulations are observed in many neurodegenerative diseases, including Alzheimer's. While these deregulations could play a key role in the development of these diseases, the study of metabolic fluxes has remained quite challenging and time-consuming. In this chapter, we describe the Seahorse XFe respirometry assay as a fundamental experimental tool to investigate the role of AMPK in controlling and modulating cell metabolic fluxes in living and intact differentiated primary neurons. The Seahorse XFe respirometry assay allows the real-time monitoring of glycolytic flux and mitochondrial respiration from different kind of cells, tissues, and isolated mitochondria. Here, we specify a protocol optimized for primary neuronal cells using several energy substrates such as glucose, pyruvate, lactate, glutamine, and ketone bodies. Nevertheless, this protocol can easily be adapted to monitor metabolic fluxes from other types of cells, tissues, or isolated mitochondria by taking into account the notes proposed for each key step of this assay.

Identification of Arabidopsis mutants with altered freezing tolerance.

PubMed

Perea-Resa, Carlos; Salinas, Julio

2014-01-01

Low temperature is an important determinant in the configuration of natural plant communities and defines the range of distribution and growth of important crops. Some plants, including Arabidopsis, have evolved sophisticated adaptive mechanisms to tolerate low and freezing temperatures. Central to this adaptation is the process of cold acclimation. By means of this process, many plants from temperate regions are able to develop or increase their freezing tolerance in response to low, nonfreezing temperatures. The identification and characterization of factors involved in freezing tolerance are crucial to understand the molecular mechanisms underlying the cold acclimation response and have a potential interest to improve crop tolerance to freezing temperatures. Many genes implicated in cold acclimation have been identified in numerous plant species by using molecular approaches followed by reverse genetic analysis. Remarkably, however, direct genetic analyses have not been conveniently exploited in their capacity for identifying genes with pivotal roles in that adaptive response. In this chapter, we describe a protocol for evaluating the freezing tolerance of both non-acclimated and cold-acclimated Arabidopsis plants. This protocol allows the accurate and simple screening of mutant collections for the identification of novel factors involved in freezing tolerance and cold acclimation.

In vitro culture of coconut (Cocos nucifera L.) zygotic embryos.

PubMed

Engelmann, Florent; Malaurie, Bernard; N'Nan, Oulo

2011-01-01

Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.

Improvement of In Vitro Date Palm Plantlet Acclimatization Rate with Kinetin and Hoagland Solution.

PubMed

Hassan, Mona M

2017-01-01

In vitro propagation of date palm Phoenix dactylifera L. is an ideal method to produce large numbers of healthy plants with specific characteristics and has the ability to transfer plantlets to ex vitro conditions at low cost and with a high survival rate. This chapter describes optimized acclimatization procedures for in vitro date palm plantlets. Primarily, the protocol presents the use of kinetin and Hoagland solution to enhance the growth of Barhee cv. plantlets in the greenhouse at two stages of acclimatization and the appropriate planting medium under shade and sunlight in the nursery. Foliar application of kinetin (20 mg/L) is recommended at the first stage. A combination between soil and foliar application of 50% Hoagland solution is favorable to plant growth and developmental parameters including plant height, leaf width, stem base diameter, chlorophyll A and B, carotenoids, and indoles. The optimum values of vegetative growth parameters during the adaptation stage in a shaded nursery are achieved using planting medium containing peat moss/perlite 2:1 (v/v), while in a sunlight nursery, clay/perlite/compost at equal ratio is the best. This protocol is suitable for large-scale production of micropropagated date palm plantlets.

Chapter 6: Ecotoxicology, Environmental Risk Assessment & Potential Impact on Human Health

EPA Science Inventory

This chapter examines potential risks posed by pharmaceuticals present in the aquatic environment to humans and aquatic life. We begin by describing the mechanisms by which pharmaceuticals enter the vertebrate body, produce effects and leave the body. Then we describe theoretical...

Chapter 5: Residential Furnaces and Boilers Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W.; Jacobson, David

The high-efficiency boiler and furnace measure produces gas heating savings resulting from installation of more energy-efficient heating equipment in a residence. Such equipment, which ranges in size from 60 kBtu/hr to 300 kBtu/hr, is installed primarily in single-family homes and multifamily buildings with individual heating systems for each dwelling unit. This protocol does not cover integrated heating and water heating units which can be used in lieu of space heating only equipment.

Histological and Morphometric Analyses for Rat Carotid Artery Balloon Injury Studies

PubMed Central

Tulis, David Anthony

2010-01-01

i. Summary Experiments aimed at analyzing the response of blood vessels to mechanical injury and ensuing remodeling responses often employ the highly characterized carotid artery balloon injury model in laboratory rats. This approach utilizes luminal insertion of a balloon embolectomy catheter into the common carotid artery with inflation and withdrawal resulting in an injury characterized by vascular endothelial cell (EC) denudation and medial wall distension. The adaptive response to this injury is typified by robust vascular smooth muscle cell (SMC) replication and migration, SMC apoptosis and necrosis, enhanced synthesis and deposition of extracellular matrix (ECM) components, partial vascular EC regeneration from the border zones, luminal narrowing and establishment of a neointima in time-dependent fashion. Evaluation of these adaptive responses to blood vessel injury can include acute and longer-term qualitative and quantitative measures including expression analyses, activity assays, immunostaining for a plethora of factors and signals, and morphometry of neointima formation and gross mural remodeling. This chapter presents a logical continuation of Chapter    in this series that offers details for performing the rat carotid artery balloon injury model in a standard laboratory setting by providing commonly used protocols for performing histological and morphometric analyses in such studies. Moreover, procedures, caveats, and considerations included in this chapter are highly relevant for alternative animal vascular physiology/pathophysiology studies and in particular those related to mechanisms of vascular injury and repair. Included in this chapter are specifics for in situ perfusion-fixation, tissue harvesting and processing for both snap-frozen and paraffin-embedded protocols, specimen embedding and sectioning, slide preparation, several standard histological staining steps, and routine morphological assessment. Included in Notes are important caveats and considerations for practical use of these methods. PMID:18287663

Pre-Emption and Precedent: The Significance of Iraq (1981) and Syria (2007) for an Israeli Response to an Iranian Nuclear Threat

DTIC Science & Technology

2011-06-10

x CHAPTER 1 INTRODUCTION ...79 1 CHAPTER 1 INTRODUCTION Background You don‘t want a messianic apocalyptic cult controlling atomic...Elli_Louka_Nuclear_Weapons_Proliferation.pdf (accessed 15 April 2011). 8 This study consists of five chapters. Chapter 1, ― Introduction ‖ announces the topic and describes its

Micropropagation of Citrus spp. by organogenesis and somatic embryogenesis.

PubMed

Chiancone, Benedetta; Germanà, Maria Antonietta

2013-01-01

Citrus spp., the largest fruit crops produced worldwide, are usually asexually propagated by cuttings or grafting onto seedling rootstocks. Most of Citrus genotypes are characterized by polyembryony due to the occurrence of adventive nucellar embryos, which lead to the production of true-to-type plants by seed germination. Tissue culture and micropropagation, in particular, are valuable alternatives to traditional propagation to obtain a high number of uniform and healthy plants in a short time and in a small space. Moreover, in vitro propagation provides a rapid system to multiply the progeny obtained by breeding programs, allows the use of monoembryonic and seedless genotypes as rootstocks, and it is very useful also for breeding and germplasm preservation.In this chapter, two protocols regarding organogenesis of a rootstock and somatic embryogenesis of a cultivar have been described.

Muskmelon embryo rescue techniques using in vitro embryo culture.

PubMed

Nuñez-Palenius, Hector Gordon; Ramírez-Malagón, Rafael; Ochoa-Alejo, Neftalí

2011-01-01

Among the major cucurbit vegetables, melon (Cucumis melo) has one of the greatest polymorphic fruit types and botanical varieties. Some melon fruits have excellent aroma, variety of flesh colors, deeper flavor, and more juice compared to other cucurbits. Despite numerous available melon cultivars, some of them are exceedingly susceptible to several diseases. The genetic background carrying the genes for tolerance and/or resistance for those diseases is found in wild melon landraces. Unfortunately, the commercial melon varieties are not able to produce viable hybrids when crossed with their wild melon counterparts. Plant tissue culture techniques are needed to surpass those genetic barriers. In vitro melon embryo rescue has played a main role to obtain viable hybrids originated from commercial versus wild melon crosses. In this chapter, an efficient and simple embryo rescue melon protocol is thoroughly described.

PET-radioimmunodetection of integrins: imaging acute colitis using a ⁶⁴Cu-labeled anti-β₇ integrin antibody.

PubMed

Dearling, Jason L J; Packard, Alan B

2012-01-01

Integrins are involved in a wide range of cell interactions. Imaging their distribution using high-resolution noninvasive techniques that are directly translatable to the clinic can provide new insights into disease processes and presents the opportunity to directly monitor new therapies. In this chapter, we describe a protocol to image, the in vivo distribution of the integrin β(7), expressed by lymphocytes recruited to and retained by the inflamed gut, using a radiolabeled whole antibody. The antibody is purified, conjugated with a bifunctional chelator for labeling with a radiometal, labeled with the positron-emitting radionuclide (64)Cu, and injected into mice for microPET studies. Mice with DSS-induced colitis were found to have higher uptake of the (64)Cu-labeled antibody in the gut than control groups.

Ballistic delivery of dyes for structural and functional studies of the nervous system

PubMed Central

Gan, Wen-Biao; Grutzendler, Jaime; Wong, Rachel O.; Lichtman, Jeff W.

2010-01-01

This chapter describes a detail protocol for rapid labeling of cells in a variety of preparations by means of particle-mediated ballistic (gene gun) delivery of fluorescent dyes. This method has been used for rapid labeling of cells with either lipid or water-soluble dyes in a variety of preparations. In particular, carbocyanine lipophilic dyes such as DiI have been used to obtain Golgi-like labeling of neurons and glia in fixed and live cell cultures, brain slices, as well as fixed post-mortem human brain. Water-soluble calcium indicators such as calcium green-1 dextran have been used to image calcium dynamics in living brain slices and retinal explants. This ballistic labeling technique is thus useful for studying the structure and function of neurons and glia in both living and fixed specimens. PMID:20147144

PERMutation Using Transposase Engineering (PERMUTE): A Simple Approach for Constructing Circularly Permuted Protein Libraries.

PubMed

Jones, Alicia M; Atkinson, Joshua T; Silberg, Jonathan J

2017-01-01

Rearrangements that alter the order of a protein's sequence are used in the lab to study protein folding, improve activity, and build molecular switches. One of the simplest ways to rearrange a protein sequence is through random circular permutation, where native protein termini are linked together and new termini are created elsewhere through random backbone fission. Transposase mutagenesis has emerged as a simple way to generate libraries encoding different circularly permuted variants of proteins. With this approach, a synthetic transposon (called a permuteposon) is randomly inserted throughout a circularized gene to generate vectors that express different permuted variants of a protein. In this chapter, we outline the protocol for constructing combinatorial libraries of circularly permuted proteins using transposase mutagenesis, and we describe the different permuteposons that have been developed to facilitate library construction.

Cord blood clinical processing, cryopreservation, and storage.

PubMed

Elmoazzen, Heidi; Holovati, Jelena L

2015-01-01

Allogeneic umbilical cord blood (UCB) hematopoietic stem cell transplantation has become a crucial advancement in the treatment for a variety of diseases including hematopoietic and non-hematopoietic malignancies, BM failure syndromes, hemoglobinopathies, and metabolic and immunodeficiency disorders. It has been well documented that the success of UCB engraftment is tied to UCB banking processes, and now there are established guidelines for standardization of collection, banking, processing, and cryopreservation for unrelated UCB units with purpose of achieving consistent production of high quality placental and UCB units for administration. In 2011, Canada's Ministry of Health has announced Canada's first national, publicly funded umbilical cord blood bank, which aims to provide altruistic donations for unrelated allogeneic hematopoietic stem cell transplant. In this chapter, we describe specific protocols for clinical processing, cryopreservation, and storage of UCB used by the Canadian Blood Services National Public Umbilical Cord Blood Bank.

Genetic transformation of carnation (Dianthus caryophylus L.).

PubMed

Nontaswatsri, Chalermsri; Fukai, Seiichi

2010-01-01

This chapter describes a rapid and efficient protocol for explant preparation and genetic transformation of carnation. Node explants from greenhouse-grown plants and leaf explants from in vitro plants are infected with Agrobacterium tumefaciens AGL0 harboring pKT3 plasmid, consisting of GUS and NPTII genes. Explant preparation is an important factor to obtain the transformed plants. The GUS-staining area was located only on the cut end of explants and only explants with a cut end close to the connecting area between node and leaf, produced transformed shoots. The cocultivation medium is also an important factor for the successful genetic transformation of carnation node and leaf explants. High genetic transformation efficiency of node and leaf explants cocultured with Agrobacterium tumefaciens was achieved when the explants were cocultivated on a filter paper soaked with water or water and acetosyringone mixture (AS).

Genetic manipulation and monitoring of autophagy in Drosophila.

PubMed

Neufeld, Thomas P

2008-01-01

Drosophila melanogaster provides a model system useful for many aspects of the study of autophagy in vivo. These include testing and validation of genes potentially involved in autophagy, discovery of novel genes through genetic screening for mutations that affect autophagy, and analysis of potential roles of autophagy in specific developmental or physiological processes. In recent years, a number of techniques and transgenic and mutant fly strains have been developed to facilitate autophagy analysis in this system. Here, protocols are described for activating or inhibiting autophagy in Drosophila, and for examining the progression of autophagy in vivo through imaging-based assays. The goal of this chapter is to provide a resource both for autophagy investigators with limited familiarity with fly genetics, as well as for experienced Drosophila biologists who wish to test for connections between autophagy and a given gene, pathway or process.

Assaying Mitochondrial Respiration as an Indicator of Cellular Metabolism and Fitness.

PubMed

Smolina, Natalia; Bruton, Joseph; Kostareva, Anna; Sejersen, Thomas

2017-01-01

Mitochondrial respiration is the most important generator of cellular energy under most circumstances. It is a process of energy conversion of substrates into ATP. The Seahorse equipment allows measuring oxygen consumption rate (OCR) in living cells and estimates key parameters of mitochondrial respiration in real-time mode. Through use of mitochondrial inhibitors, four key mitochondrial respiration parameters can be measured: basal, ATP production-linked, maximal, and proton leak-linked OCR. This approach requires application of mitochondrial inhibitors-oligomycin to block ATP synthase, FCCP-to make the inner mitochondrial membrane permeable for protons and allow maximum electron flux through the electron transport chain, and rotenone and antimycin A-to inhibit complexes I and III, respectively. This chapter describes the protocol of OCR assessment in the culture of primary myotubes obtained upon satellite cell fusion.

The stem cell laboratory: design, equipment, and oversight.

PubMed

Wesselschmidt, Robin L; Schwartz, Philip H

2011-01-01

This chapter describes some of the major issues to be considered when setting up a laboratory for the culture of human pluripotent stem cells (hPSCs). The process of establishing a hPSC laboratory can be divided into two equally important parts. One is completely administrative and includes developing protocols, seeking approval, and establishing reporting processes and documentation. The other part of establishing a hPSC laboratory involves the physical plant and includes design, equipment and personnel. Proper planning of laboratory operations and proper design of the physical layout of the stem cell laboratory so that meets the scope of planned operations is a major undertaking, but the time spent upfront will pay long-term returns in operational efficiency and effectiveness. A well-planned, organized, and properly equipped laboratory supports research activities by increasing efficiency and reducing lost time and wasted resources.

Traffic Pattern Detection Using the Hough Transformation for Anomaly Detection to Improve Maritime Domain Awareness

DTIC Science & Technology

2013-12-01

Programming code in the Python language used in AIS data preprocessing is contained in Appendix A. The MATLAB programming code used to apply the Hough...described in Chapter III is applied to archived AIS data in this chapter. The implementation of the method, including programming techniques used, is...is contained in the second. To provide a proof of concept for the algorithm described in Chapter III, the PYTHON programming language was used for

NASA directives: Master list and index

NASA Technical Reports Server (NTRS)

1994-01-01

This Handbook sets forth in two parts the following information for the guidance of users of the NASA Management Directives System. Chapter 1 contains introductory information material on how to use this Handbook. Chapter 2 is a complete master list of Agency-wide management directives, describing each directive by type, number, effective date, expiration date, title, and organization code of the office responsible for the directive. Chapter 3 includes a consolidated numerical list of all delegations of authority and a breakdown of such delegation by the office of Installation to which special authority is assigned. Chapter 4 sets forth a consolidated list of all NASA Handbooks (NHB's) and important footnotes covering the control and ordering of such documents. Chapter 5 is a consolidated list of NASA management directives applicable to the Jet Propulsion Laboratory. Chapter 6 is a consolidated list of NASA management directives published in the code of Federal Regulations. Complementary manuals to the NASA Management Directives System are described in Chapter 7. Part B contains an in-depth alphabetical index to all NASA management directives other than Handbooks.

A Guide for Using the Internet.

ERIC Educational Resources Information Center

Brown, Herb

This manual provides an instructional overview of Internet resources with student exercises. The work consists of nine chapters: (1) Introduction to the Internet; (2) How to Access the Internet; (3) Electronic Mail; (4) Gopher; (5) File Transfer Protocol-FTP; (6) Newsgroups & Newsreaders; (7) Telnet & TN3270; (8) World Wide Web; and (9)…

Jobs in Public Service. Job Family Series.

ERIC Educational Resources Information Center

Science Research Associates, Inc., Chicago, IL.

The booklet describes jobs in public service. The introductory chapter provides a definition and overview of public service work. Following chapters describe various levels of public service including: (1) neighborhood workers--letter carriers and postal employees, fire fighters, police officers, service repair workers, and bus drivers; (2) city…

User's guide [Chapter 3

Treesearch

Nicholas L. Crookston; Donald C. E. Robinson; Sarah J. Beukema

2003-01-01

The Fire and Fuels Extension (FFE) to the Forest Vegetation Simulator (FVS) simulates fuel dynamics and potential fire behavior over time, in the context of stand development and management. This chapter presents the model's options, provides annotated examples, describes the outputs, and describes how to use and apply the model.

CoP Sensing Framework on Web-Based Environment

NASA Astrophysics Data System (ADS)

Mustapha, S. M. F. D. Syed

The Web technologies and Web applications have shown similar high growth rate in terms of daily usages and user acceptance. The Web applications have not only penetrated in the traditional domains such as education and business but have also encroached into areas such as politics, social, lifestyle, and culture. The emergence of Web technologies has enabled Web access even to the person on the move through PDAs or mobile phones that are connected using Wi-Fi, HSDPA, or other communication protocols. These two phenomena are the inducement factors toward the need of building Web-based systems as the supporting tools in fulfilling many mundane activities. In doing this, one of the many focuses in research has been to look at the implementation challenges in building Web-based support systems in different types of environment. This chapter describes the implementation issues in building the community learning framework that can be supported on the Web-based platform. The Community of Practice (CoP) has been chosen as the community learning theory to be the case study and analysis as it challenges the creativity of the architectural design of the Web system in order to capture the presence of learning activities. The details of this chapter describe the characteristics of the CoP to understand the inherent intricacies in modeling in the Web-based environment, the evidences of CoP that need to be traced automatically in a slick manner such that the evidence-capturing process is unobtrusive, and the technologies needed to embrace a full adoption of Web-based support system for the community learning framework.

Chapter Four: Discursive Resources

ERIC Educational Resources Information Center

Young, Richard F.

2008-01-01

In this chapter, the focus of attention moves from the contexts described in chapter 3 to the verbal, nonverbal, and interactional resources that participants employ in discursive practices. These resources are discussed within the frame of participation status and participation framework proposed by Goffman. Verbal resources employed by…

The School District Organization Handbook.

ERIC Educational Resources Information Center

California State Dept. of Education, Sacramento.

This handbook describes procedures for school-district reorganization in California. Following the introductory chapter, chapter 2 offers a historical overview of school-district reorganization in California. Chapters 3 and 4 outline the organization and responsibilities of the county committee and the role and responsibilities of the State Board…

BOOK REVIEW: The Illustrated Wavelet Transform Handbook: Introductory Theory and Applications in Science, Engineering, Medicine and Finance

NASA Astrophysics Data System (ADS)

Ng, J.; Kingsbury, N. G.

2004-02-01

This book provides an overview of the theory and practice of continuous and discrete wavelet transforms. Divided into seven chapters, the first three chapters of the book are introductory, describing the various forms of the wavelet transform and their computation, while the remaining chapters are devoted to applications in fluids, engineering, medicine and miscellaneous areas. Each chapter is well introduced, with suitable examples to demonstrate key concepts. Illustrations are included where appropriate, thus adding a visual dimension to the text. A noteworthy feature is the inclusion, at the end of each chapter, of a list of further resources from the academic literature which the interested reader can consult. The first chapter is purely an introduction to the text. The treatment of wavelet transforms begins in the second chapter, with the definition of what a wavelet is. The chapter continues by defining the continuous wavelet transform and its inverse and a description of how it may be used to interrogate signals. The continuous wavelet transform is then compared to the short-time Fourier transform. Energy and power spectra with respect to scale are also discussed and linked to their frequency counterparts. Towards the end of the chapter, the two-dimensional continuous wavelet transform is introduced. Examples of how the continuous wavelet transform is computed using the Mexican hat and Morlet wavelets are provided throughout. The third chapter introduces the discrete wavelet transform, with its distinction from the discretized continuous wavelet transform having been made clear at the end of the second chapter. In the first half of the chapter, the logarithmic discretization of the wavelet function is described, leading to a discussion of dyadic grid scaling, frames, orthogonal and orthonormal bases, scaling functions and multiresolution representation. The fast wavelet transform is introduced and its computation is illustrated with an example using the Haar wavelet. The second half of the chapter groups together miscellaneous points about the discrete wavelet transform, including coefficient manipulation for signal denoising and smoothing, a description of Daubechies’ wavelets, the properties of translation invariance and biorthogonality, the two-dimensional discrete wavelet transforms and wavelet packets. The fourth chapter is dedicated to wavelet transform methods in the author’s own specialty, fluid mechanics. Beginning with a definition of wavelet-based statistical measures for turbulence, the text proceeds to describe wavelet thresholding in the analysis of fluid flows. The remainder of the chapter describes wavelet analysis of engineering flows, in particular jets, wakes, turbulence and coherent structures, and geophysical flows, including atmospheric and oceanic processes. The fifth chapter describes the application of wavelet methods in various branches of engineering, including machining, materials, dynamics and information engineering. Unlike previous chapters, this (and subsequent) chapters are styled more as literature reviews that describe the findings of other authors. The areas addressed in this chapter include: the monitoring of machining processes, the monitoring of rotating machinery, dynamical systems, chaotic systems, non-destructive testing, surface characterization and data compression. The sixth chapter continues in this vein with the attention now turned to wavelets in the analysis of medical signals. Most of the chapter is devoted to the analysis of one-dimensional signals (electrocardiogram, neural waveforms, acoustic signals etc.), although there is a small section on the analysis of two-dimensional medical images. The seventh and final chapter of the book focuses on the application of wavelets in three seemingly unrelated application areas: fractals, finance and geophysics. The treatment on wavelet methods in fractals focuses on stochastic fractals with a short section on multifractals. The treatment on finance touches on the use of wavelets by other authors in studying stock prices, commodity behaviour, market dynamics and foreign exchange rates. The treatment on geophysics covers what was omitted from the fourth chapter, namely, seismology, well logging, topographic feature analysis and the analysis of climatic data. The text concludes with an assortment of other application areas which could only be mentioned in passing. Unlike most other publications in the subject, this book does not treat wavelet transforms in a mathematically rigorous manner but rather aims to explain the mechanics of the wavelet transform in a way that is easy to understand. Consequently, it serves as an excellent overview of the subject rather than as a reference text. Keeping the mathematics to a minimum and omitting cumbersome and detailed proofs from the text, the book is best-suited to those who are new to wavelets or who want an intuitive understanding of the subject. Such an audience may include graduate students in engineering and professionals and researchers in engineering and the applied sciences.

Study of Orbitally Excited $$B_{(s)}$$ Mesons and Evidence for a New $$B\\pi$$ Resonance with the CDF II Detector

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

Kambeitz, Manuel

This thesis presents an analysis of excited states of B0, B+ and B0 s mesons, decaying to B mesons while emitting a pion or kaon. They are reconstructed from their decay products and a selection is performed to discard wrongly reconstructed B(s) mesons with the multivariate analysis software NeuroBayes, as described in chapter 5. In the training process, the sPlot method and measured and simulated data are used. Chapter 6 describes how the properties of excited B(s) are determined by an unbinned maximum likelihood t to their mass spectra. The systematic uncertainties determined in this analysis are described in chaptermore » 7. The results of this thesis are presented in chapter 8 and a conclusion is given in chapter 9. The results shown in this thesis have been published before in [1].« less

Total Quality Management in Education.

ERIC Educational Resources Information Center

Johnson, James H.

1993-01-01

Ways to apply the concepts and processes of Total Quality Management (TQM) to education are discussed in this document. Following the introduction and the preface, chapter 1 provides a historical overview and describes the four cornerstones of TQM--an understanding of systems, psychology, knowledge, and statistics. Chapter 2 describes some of the…

Soft Paths: How To Enjoy the Wilderness without Harming It.

ERIC Educational Resources Information Center

Hampton, Bruce; Cole, David

This outdoor-education book describes methods of hiking and camping that minimize the human impact upon the natural environment. Each chapter offers the rationale behind recommended practices, based on the best scientific research on recreational impact. The first chapter, "The Case for Minimum Impact," describes increasing tourist use…

Food Rights Handbook.

ERIC Educational Resources Information Center

Children's Foundation, Washington, DC.

This booklet, which describes federal food assistance programs, is designed to help large families, families on small budgets, and elderly people on fixed incomes get more food for less money. The book is divided into four chapters: Kids, Women and Children, Families, and Senior Citizens. Each chapter describes in detail the food assistance…

Pre-School Education and Care.

ERIC Educational Resources Information Center

Haystead, J.; And Others

Carried out in Scotland, the project described in this book examined what Scottish parents want from preschool education and why. The first chapter describes the present situation of preschool education and how it serves different kinds of parents. Chapter II discusses the kinds of preschool provision desired by parents living in three different…

New and Emerging Occupations Project. Final Report.

ERIC Educational Resources Information Center

Southeast Community Coll., Lincoln, NE.

This document reports a project to define, identify, and describe new and emerging occupations in Nebraska and disseminate that information to vocational education program and curriculum planners. Chapter 1 describes the background, problem, and purpose of the project. Chapter 2 sets forth the objectives and discusses procedures for developing a…

To Teach, To Love.

ERIC Educational Resources Information Center

Stuart, Jesse

In this book, the Kentucky novelist, poet, and teacher Jesse Stuart describes his experiences as student and teacher. Chapter 1 recounts the influence of his father, who instilled the principle of learning through play and the value of imagination and memory, and catalogs young Jesse's teachers at Plum Grove School. Chapter 2 describes his high…

Towards Universalization of Primary Education in Asia and the Pacific: Country Studies--Nepal.

ERIC Educational Resources Information Center

United Nations Educational, Scientific, and Cultural Organization, Bangkok (Thailand). Regional Office for Education in Asia and the Pacific.

The four major chapters of this study describe education in Nepal, analyze the progress of primary education in that country, describe national policy and planning for primary education, and report significant developments and programs. Chapter One concerns the historical background, present structure of education, primary education…

Effects of climate change on ecological disturbances [Chapter 8

Treesearch

Danielle M. Malesky; Barbara J. Bentz; Gary R. Brown; Andrea R. Brunelle; John M. Buffington; Linda M. Chappell; R. Justin DeRose; John C. Guyon; Carl L. Jorgensen; Rachel A. Loehman; Laura L. Lowrey; Ann M. Lynch; Marek Matyjasik; Joel D. McMillin; Javier E. Mercado; Jesse L. Morris; Jose F. Negron; Wayne G. Padgett; Robert A. Progar; Carol B. Randall

2018-01-01

This chapter describes disturbance regimes in the Intermountain Adaptation Partnership (IAP) region, and potential shifts in these regimes as a consequence of observed and projected climate change. The term "disturbance regime" describes the general temporal and spatial characteristics of a disturbance agent (e.g., insects, disease, fire, weather, human...

The Use of Technology in Adult Literacy Programs.

ERIC Educational Resources Information Center

Anderson, Jonathan; And Others

This document describes the use of educational technology (including radio, television, computers, telephones, satellites, and optical laser discs) in adult literacy programs in Australia. Chapter 1 describes the scope of the study that resulted in the document and defines both literacy and educational technology. Chapter 2 contains a generalized…

Multisectoral Strategies for Advancing Girls' Education: Principles and Practice. SAGE Technical Report.

ERIC Educational Resources Information Center

Williams, Howard

This paper describes issues, experiences, and strategies used in developing successful multisectoral partnerships to advance girls' education, using Guinea and Morocco as examples. Chapter 1 introduces the issue, discussing barriers to girls' education and describing the multisectoral response to interrelated barriers. Chapter 2 defines the…

Minneapolis Multi-Ethnic Curriculum Project--Migration Unit.

ERIC Educational Resources Information Center

Minneapolis Public Schools, Minn. Dept. of Intergroup Education.

The student booklet presents short chapters illustrating the migration unit of the Minneapolis Multi-Ethnic Curriculum Project for secondary schools. Sixteen brief chapters describe migration, immigration, and emigration in the United States. The first six chapters offer first person accounts of immigrants from Norway, Korea, Egypt, Hitler's…

Energy: Options and Issues.

ERIC Educational Resources Information Center

Walker, Harry O.

This book is intended to provide basic information about energy. The first three chapters describe energy supply and demand, uses and sources, and common energy terms. The next two chapters explain environmental and biological effects of energy systems. Twelve chapters that follow outline past history and technological knowledge of the following…

What Research Tells the Coach About Sprinting.

ERIC Educational Resources Information Center

Dintiman, George B.

This booklet on sprinting is divided into four chapters. Chapter 1 "Introduction," provides an analysis of the 100-meter dash, summarizes world records, and discusses the reliability of timing the sprint race. Chapter 2, "Describing the Sprinter," discusses the following topics: anatomical characteristics, flexibility, reaction, strength/power,…

Transport Protocols for Wireless Mesh Networks

NASA Astrophysics Data System (ADS)

Eddie Law, K. L.

Transmission control protocol (TCP) provides reliable connection-oriented services between any two end systems on the Internet. With TCP congestion control algorithm, multiple TCP connections can share network and link resources simultaneously. These TCP congestion control mechanisms have been operating effectively in wired networks. However, performance of TCP connections degrades rapidly in wireless and lossy networks. To sustain the throughput performance of TCP connections in wireless networks, design modifications may be required accordingly in the TCP flow control algorithm, and potentially, in association with other protocols in other layers for proper adaptations. In this chapter, we explain the limitations of the latest TCP congestion control algorithm, and then review some popular designs for TCP connections to operate effectively in wireless mesh network infrastructure.

In Vitro Analysis of the Role of Schwann Cells on Axonal Degeneration and Regeneration Using Sensory Neurons from Dorsal Root Ganglia.

PubMed

López-Leal, Rodrigo; Diaz, Paula; Court, Felipe A

2018-01-01

Sensory neurons from dorsal root ganglion efficiently regenerate after peripheral nerve injuries. These neurons are widely used as a model system to study degenerative mechanisms of the soma and axons, as well as regenerative axonal growth in the peripheral nervous system. This chapter describes techniques associated to the study of axonal degeneration and regeneration using explant cultures of dorsal root ganglion sensory neurons in vitro in the presence or absence of Schwann cells. Schwann cells are extremely important due to their involvement in tissue clearance during axonal degeneration as well as their known pro-regenerative effect during regeneration in the peripheral nervous system. We describe methods to induce and study axonal degeneration triggered by axotomy (mechanical separation of the axon from its soma) and treatment with vinblastine (which blocks axonal transport), which constitute clinically relevant mechanical and toxic models of axonal degeneration. In addition, we describe three different methods to evaluate axonal regeneration using quantitative methods. These protocols constitute a valuable tool to analyze in vitro mechanisms associated to axonal degeneration and regeneration of sensory neurons and the role of Schwann cells in these processes.

Measurements of CP asymmetries and branching fractions of two-body charmless decays of B 0 and B$$0\\atop{s}$$ mesons

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

Morello, Michael Joseph

2007-12-19

The thesis is organized as follows: Chapter 1 describes the theoretical framework of non-leptonic Bmore » $$0\\atop{s}$$ → H +h' - decays, with a simple overview of the CP violation mechanism within the Standard Model and of the most used phenomenological approaches in the evaluation of strong interaction contributions. The chapter contains also a review of the theoretical expectations and the current experimental measurements along with a discussion about the importance of studying such decays. Chapter 2 contains a general description of the Tevatron collider and of the CDF II detector. Chapter 3 is devoted to the description of the data sample used for the measurement and the method used in extracting the signal from the background. Particular attention is dedicated to the on-line trigger selection, which is crucial to collect a sample enriched in B$$0\\atop{s}$$ → h +h' - decays. Chapter 4 shows how the information from kinematics and particle identification was used to achieve a statistical discrimination amongst modes to extract individual measurements. The available resolutions in mass or in particle identification are separately insufficient for an event-by-event separation of B$$0\\atop{s}$$ → h +h' - modes. The choice of observables and the technique used to combine them is an important and innovative aspect of the analysis described in this thesis. Chapter 5 is devoted to the accurate determination of the invariant mass lineshape. This is a crucial ingredient for resolving overlapping mass peaks. This chapter details all resolution effects with particular attention at the tails due to the emission of low-energy photons from charged kaons and pions in the final state (FSR). For the first time the effect of FSR has been accurately accounted for in a CDF analysis. Chapter 6 describes how kinematic and PID information, discussed in chap. 4 and chap. 5 were combined in a maximum Likelihood fit to statistically determine the composition of the B$$0\\atop{s}$$ → h +h' - sample. This kinematics-PID combined fit has been developed and performed for the first time at CDF in the analysis presented in this thesis and this methodology was later inherited by several other analyses. Chapter 7 is devoted to the study of the isolation variable, which is a crucial handle to enhance the signal-to-background ratio in the off-line selection. It exploits the property that the b-hadrons tend to carry a larger fraction of the transverse momentum of the particles produced in the fragmentation, with respect to lighter hadrons. Since the simulators do not accurately reproduce the fragmentation processes, this chapter is devoted to the study of the control data sample of B$$0\\atop{s}$$ → J/ΨX decays to probe the characteristics of this variable. Chapter 8 describes an innovative procedure used to optimize the selection to minimize the statistical uncertainty on the quantities one wishes to measure. The procedure is based on the fit of composition described in chap. 6. Chapter 9 reports the results of the fit of composition described in chap. 6 and the cross-checks performed to verify the goodness of the fit of composition. In order to translate the parameters returned from the fit into physics measurements the relative efficiency corrections between the various decay modes need to be applied. Chapter 10 is devoted to the description of these corrections. Chapter 11 describes the measurement of the detector-induced charge asymmetry between positively and negatively charged kaons and pions, due to their different probability of strong interaction in the tracker material using the real data. This allows to extract the acceptance correction factor for the CP asymmetries measurement without any external inputs from the simulation, and to perform a powerful check of whole analysis. Chapter 12 describes the main sources of systematic uncertainties and the method used to evaluate the significance of the results on rare modes. The final results of the measurements and their interpretation are discussed in chap. 13.« less

Novel chemistries and materials for grid-scale energy storage: Quinones and halogen catalysis

NASA Astrophysics Data System (ADS)

Huskinson, Brian Thomas

In this work I describe various approaches to electrochemical energy storage at the grid-scale. Chapter 1 provides an introduction to energy storage and an overview of the history and development of flow batteries. Chapter 2 describes work on the hydrogen-chlorine regenerative fuel cell, detailing its development and the record-breaking performance of the device. Chapter 3 dives into catalyst materials for such a fuel cell, focusing on ruthenium oxide based alloys to be used as chlorine redox catalysts. Chapter 4 introduces and details the development of a performance model for a hydrogen-bromine cell. Chapter 5 delves into the more recent work I have done, switching to applications of quinone chemistries in flow batteries. It focuses on the pairing of one particular quinone (2,7-anthraquinone disulfonic acid) with bromine, and highlights the promising performance characteristics of a device based on this type of chemistry.

Where Have All the Sparrows Gone?: Rethinking Chapter I Services.

ERIC Educational Resources Information Center

Standerford, N. Suzanne

1993-01-01

Describes a restructured Chapter 1 program that provided supplemental support within the students' classrooms to raise expectations for Chapter 1 students' performance. Notes that expectations of teachers and students increased; portfolios of student work showed steady improvement in reading comprehension and writing ability; and standardized test…

Increasing Student Learning through Multimedia Projects.

ERIC Educational Resources Information Center

Simkins, Michael; Cole, Karen; Tavalin, Fern; Means, Barbara

This book discusses enhancing student achievement through project-based learning with multimedia. Chapter 1 describes project-based multimedia learning. Chapter 2 presents a multimedia primer, including the five basic types of media objects (i.e., images, text, sound, motion, and interactivity). Chapter 3 addresses making a real-world connection,…

Jobs in Manufacturing. Job Family Series.

ERIC Educational Resources Information Center

Science Research Associates, Inc., Chicago, IL.

The instructional booklet explores various occupations in the occupational cluster of manufacturing. In the first chapter, the student is briefly introduced to the occupational cluster concept and to the general area of manufacturing. Chapter 2 describes jobs involved in the production of materials and processing of goods. Chapter 3 discusses…

Ecology, Ecosystem Management and Biology Teaching. Biology and Human Welfare.

ERIC Educational Resources Information Center

Spellerberg, Ian F.; Pritchard, Alan J.

This six-chapter document (part of a series on biology and human welfare) focuses on ecology, ecosystem management, and biology teaching. Chapter 1 discusses the basic elements of ecology (considering organisms and their environment, populations, and communities and ecosystems). Chapter 2 describes several aspects of human ecology and resources…

The Homeless in Contemporary Society.

ERIC Educational Resources Information Center

Bingham, Richard D.; And Others

This book consists of 15 chapters on understanding and helping the homeless. The first seven chapters present the "new" homeless in historical context and describe this population and its situation. The remaining eight chapters discuss policy and program options of the government and other organizations in attempting to alleviate the problems of…

State of the Environment: An Assessment at Mid-Decade.

ERIC Educational Resources Information Center

Conservation Foundation, Washington, DC.

This report is divided into two parts. The three chapters in part 1 describe environmental conditions and trends. Chapter 1 deals with underlying trends, primarily population growth and economic factors. Chapter 2 covers environmental contaminants, including toxic substances, hazardous waste, air and water pollutants, and overall waste production.…

NASA directives master list and index

NASA Technical Reports Server (NTRS)

1995-01-01

This handbook sets forth in two parts, Master List of Management Directives and Index to NASA Management Directives, the following information for the guidance of users of the NASA Management Directives System. Chapter 1 contains introductory information material on how to use this handbook. Chapter 2 is a complete master list of agencywide management directives, describing each directive by type, number, effective date, expiration date, title, and organization code of the office responsible for the directive. Chapter 3 includes a consolidated numerical list of all delegations of authority and a breakdown of such delegation by the office or center to which special authority is assigned. Chapter 4 sets forth a consolidated list of all NASA handbooks (NHB's) and important footnotes covering the control and ordering of such documents. Chapter 5 is a consolidated list of NASA management directives applicable to the Jet Propulsion Laboratory. Chapter 6 is a consolidated list of NASA regulations published in the Code of Federal Regulations. Chapter 7 is a consolidated list of NASA regulations published in Title 14 of the Code of Federal Regulations. Complementary manuals to the NASA Management Directives System are described in Chapter 8. The second part contains an in depth alphabetical index to all NASA management directives other than handbooks, most of which are indexed by titles only.

Procedures and Standards Handbook. Version 3.0. What Works Clearinghouse

ERIC Educational Resources Information Center

What Works Clearinghouse, 2014

2014-01-01

This "What Works Clearinghouse Procedures and Standards Handbook (Version 3.0)" provides a detailed description of the standards and procedures of the What Works Clearinghouse (WWC). The remaining chapters of this Handbook are organized to take the reader through the basic steps that the WWC uses to develop a review protocol, identify…

Planning crops and developing propagation protocols [Chapter 3

Treesearch

Douglass F. Jacobs; Kim M. Wilkinson

2009-01-01

Crop planning is an important but often neglected aspect of successful nursery management. Crop planning enables proper scheduling of the necessary time, materials, labor, and space to produce crops. Many painstaking details, such as the careful design of nursery facilities; working with clients; collecting and propagating seeds and cuttings; and making improvements in...

By-Laws | Center for Cancer Research

Cancer.gov

The NIH Fellows Editorial Board (FEB) offers a free and confidential editorial service for NIH fellows. The Board members edit manuscripts, abstracts, book chapters, clinical protocols, and grant submissions and offer suggestions for grammar, structure, and style, but not for scientific content. The Board provides valuable training and editorial experience to its members for

Still Teaching Ourselves.

ERIC Educational Resources Information Center

Leistico, Agnes

This book is the sequel to "I Learn Better By Teaching Myself," which argued that parents need to follow their personal parenting instincts and to trust that their students know what is best for themselves. Chapter 1 describes how the author's family became involved in home schooling. The second chapter describes how they set standards in the form…

The changing southwest

USGS Publications Warehouse

David M. Theobald,; William Travis,; Drummond, Mark A.; Eric Gordon,; Michelle Betsill,

2013-01-01

This chapter describes important geographical and socio-economic characteristics and trends in the Southwest—such as population and economic growth and changes in land ownership, land use, and land cover—that provide the context for how climate change will likely affect the Southwest. The chapter also describes key laws and institutions relevant to adaptive management of resources.

Solar Physics in the Space Age.

ERIC Educational Resources Information Center

Dittmer, Phil D.; And Others

This amply illustrated booklet provides a physical description of the sun as well as present and future tasks for solar physics study. The first chapter, an introduction, describes the history of solar study, solar study in space, and the relevance of solar study. The second chapter describes the five heliographic domains including the interior,…

Can Ethics Be Taught? Perspectives, Challenges, and Approaches at Harvard Business School.

ERIC Educational Resources Information Center

Piper, Thomas R.; And Others

This book describes in five chapters how the Harvard Business School has redeveloped its curriculum to place leadership, ethics, and corporate responsibility at the center of its mission. Chapter 1, "Rediscovery of Purpose: The Genesis of the Leadership, Ethics, and Corporate Responsibility Initiative," (Thomas R. Piper) describes the…

Instructional Technology: A Systematic Approach to Remedying Everyday Educational/Classroom Challenge.

ERIC Educational Resources Information Center

Williams, W. Wes; Hasazi, Susan E.

The authors describe program organization and teaching/learning procedures which can be employed in the remediation of mildly to severely handicapped students. Following an introductory chapter, Chapter 2 describes ways to manage educational resources such as time, aides, support staff, and student teachers to maximize learning. A brief third…

A Forest Products Technology Program for Washington Community Colleges. Final Report.

ERIC Educational Resources Information Center

Grotefend, Robert T.

This report describes a project to develop a forest product technology curriculum that prepares students at the technical level, junior management, to work in the manufacturing of wood and bark into lumber, plywood, particle board, laminates, and pulp chips. Chapter 1 describes project objectives and procedures followed. In chapter 2 the history…

Chapter 22: Compressed Air Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W; Benton, Nathanael; Burns, Patrick

Compressed-air systems are used widely throughout industry for many operations, including pneumatic tools, packaging and automation equipment, conveyors, and other industrial process operations. Compressed-air systems are defined as a group of subsystems composed of air compressors, air treatment equipment, controls, piping, pneumatic tools, pneumatically powered machinery, and process applications using compressed air. A compressed-air system has three primary functional subsystems: supply, distribution, and demand. Air compressors are the primary energy consumers in a compressed-air system and are the primary focus of this protocol. The two compressed-air energy efficiency measures specifically addressed in this protocol are: High-efficiency/variable speed drive (VSD) compressormore » replacing modulating, load/unload, or constant-speed compressor; and Compressed-air leak survey and repairs. This protocol provides direction on how to reliably verify savings from these two measures using a consistent approach for each.« less

SeaWiFS Postlaunch Calibration and Validation Analyses

NASA Technical Reports Server (NTRS)

Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor); McClain, Charles R.; Ainsworth, Ewa J.; Barnes, Robert A.; Eplee, Robert E., Jr.; Patt, Frederick S.; Robinson, Wayne D.; Wang, Menghua; Bailey, Sean W.

2000-01-01

The effort to resolve data quality issues and improve on the initial data evaluation methodologies of the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Project was an extensive one. These evaluations have resulted, to date, in three major reprocessings of the entire data set where each reprocessing addressed the data quality issues that could be identified up to the time of each reprocessing. The number of chapters (21) needed to document this extensive work in the SeaWiFS Postlaunch Technical Report Series requires three volumes. The chapters in Volumes 9, 10, and 11 are in a logical order sequencing through sensor calibration, atmospheric correction, masks and flags, product evaluations, and bio-optical algorithms. The first chapter of Volume 9 is an overview of the calibration and validation program, including a table of activities from the inception of the SeaWiFS Project. Chapter 2 describes the fine adjustments of sensor detector knee radiances, i.e., radiance levels where three of the four detectors in each SeaWiFS band saturate. Chapters 3 and 4 describe the analyses of the lunar and solar calibration time series, respectively, which are used to track the temporal changes in radiometric sensitivity in each band. Chapter 5 outlines the procedure used to adjust band 7 relative to band 8 to derive reasonable aerosol radiances in band 7 as compared to those in band 8 in the vicinity of Lanai, Hawaii, the vicarious calibration site. Chapter 6 presents the procedure used to estimate the vicarious calibration gain adjustment factors for bands 1-6 using the waterleaving radiances from the Marine Optical Buoy (MOBY) offshore of Lanai. Chapter 7 provides the adjustments to the coccolithophore flag algorithm which were required for improved performance over the prelaunch version. Chapter 8 is an overview of the numerous modifications to the atmospheric correction algorithm that have been implemented. Chapter 9 describes the methodology used to remove artifacts of sun glint contamination for portions of the imagery outside the sun glint mask. Finally, Chapter 10 explains a modification to the ozone interpolation method to account for actual time differences between the SeaWiFS and Total Ozone Mapping Spectrometer (TOMS) orbits.

Prototype Engineered Barrier System Field Test (PEBSFT); Final report

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

Ramirez, A.L.; Buscheck, T.; Carlson, R.

1991-08-01

This final report represents a summary of data and interpretations obtained from the Prototype Engineered Barrier System Field Test (PEBSFT) performed in G-Tunnel within the Nevada Test Site. The PEBSFT was conducted to evaluate the applicability of measurement techniques, numerical models, and procedures developed for future field tests that will be conducted in the Exploratory Studies Facilities (ESF) at Yucca Mountain. The primary objective of the test was to provide a basis for determining whether tests planned for the ESF have the potential to be successful. Chapter 1 on high frequency electromagnetic tomography discusses the rock mass electromagnetic permittivity andmore » attenuation rate changes that were measured to characterize the water distribution in the near field of a simulated waste container. The data are used to obtain quantitative estimates of how the moisture content in the rock mass changes during heating and to infer properties of the spatial variability of water distribution, leading to conclusions about the role of fractures in the system. Chapter 2 discusses the changes in rock moisture content detected by the neutron logging probe. Chapter 3 permeability tests discusses the characterization of the in-situ permeability of the fractured tuff around the borehole. The air permeability testing apparatus, the testing procedures, and the data analysis are presented. Chapter 4 describes the moisture collection system installed in the heater borehole to trap and measure the moisture volumes. Chapter 5 describes relative humidity measurements made with the thermocouple psychrometer and capacitance sensors. Chapter 6 discusses gas pressure measurements in the G-Tunnel, addressing the calibration and installation of piezoresistive-gaged transducers. Chapter 7 describes the calibration and installation of thermocouples for temperature measurements. Chapter 8 discusses the results of the PEBSFT.« less

Isolation and culture of human multipotent stromal cells from the pancreas.

PubMed

Seeberger, Karen L; Eshpeter, Alana; Korbutt, Gregory S

2011-01-01

Mesenchymal stem cells, also termed multipotent mesenchymal stromal cells (MSCs), can be isolated from most adult tissues. Although the exact origin of MSCs expanded from the human pancreas has not been resolved, we have developed protocols to isolate and expand MSCs from human pancreatic tissue that remains after islet procurement. Similar to techniques used to isolate MSCs from bone marrow, pancreatic MSCs are isolated based on their cell adherence, expression of several cell surface antigens, and multilineage differentiation. The protocols for isolating, characterizing, and differentiating MSCs from the pancreas are presented in this chapter.

Forces of Commonly Used Chiropractic Techniques for Children: A Review of the Literature.

PubMed

Todd, Angela J; Carroll, Matthew T; Mitchell, Eleanor K L

2016-01-01

The purpose of this study is to review the available literature that describes forces of the most commonly used chiropractic techniques for children. Review of the English-language literature using search terms Chiropract* and technique, protocol, or approach in databases PubMed, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine, and Index to Chiropractic Literature and direct contact with authors of articles and book chapters. Eleven articles that discussed the 7 most commonly used pediatric chiropractic techniques and the forces applied were identified. Chiropractic techniques reviewed described forces that were modified based on the age of the patient. Force data for mechanically assisted devices were varied, with the minimum force settings for some devices outside the age-specific safe range recommended in the literature when not modified in some way. This review found that technique selection and application by chiropractors treating infants and young children are typically modified in force and speed to suit the age and development of the child. Copyright © 2016. Published by Elsevier Inc.

A Recessive Pollination Control System for Wheat Based on Intein-Mediated Protein Splicing.

PubMed

Gils, Mario

2017-01-01

A transgene-expression system for wheat that relies on the complementation of inactive precursor protein fragments through a split-intein system is described. The N- and C-terminal fragments of a barnase gene from Bacillus amyloliquifaciens were fused to intein sequences from Synechocystis sp. and transformed into wheat plants. Upon translation, both barnase fragments are assembled by an autocatalytic intein-mediated trans-splicing reaction, thus forming a cytotoxic enzyme. This chapter focuses on the use of introns and flexible polypeptide linkers to foster the expression of a split-barnase expression system in plants. The methods and protocols that were employed with the objective to test the effects of such genetic elements on transgene expression and to find the optimal design of expression vectors for use in wheat are provided. Split-inteins can be used to form an agriculturally important trait (male sterility) in wheat plants. The use of this principle for the production of hybrid wheat seed is described. The suggested toolbox will hopefully be a valuable contribution to future optimization strategies in this commercially important crop.

Genotyping and Molecular Identification of Date Palm Cultivars Using Inter-Simple Sequence Repeat (ISSR) Markers.

PubMed

Ayesh, Basim M

2017-01-01

Molecular markers are credible for the discrimination of genotypes and estimation of the extent of genetic diversity and relatedness in a set of genotypes. Inter-simple sequence repeat (ISSR) markers rapidly reveal high polymorphic fingerprints and have been used frequently to determine the genetic diversity among date palm cultivars. This chapter describes the application of ISSR markers for genotyping of date palm cultivars. The application involves extraction of genomic DNA from the target cultivars with reliable quality and quantity. Subsequently the extracted DNA serves as a template for amplification of genomic regions flanked by inverted simple sequence repeats using a single primer. The similarity of each pair of samples is measured by calculating the number of mono- and polymorphic bands revealed by gel electrophoresis. Matrices constructed for similarity and genetic distance are used to build a phylogenetic tree and cluster analysis, to determine the molecular relatedness of cultivars. The protocol describes 3 out of 9 tested primers consistently amplified 31 loci in 6 date palm cultivars, with 28 polymorphic loci.

3D-Printing Composite Polycaprolactone-Decellularized Bone Matrix Scaffolds for Bone Tissue Engineering Applications.

PubMed

Rindone, Alexandra N; Nyberg, Ethan; Grayson, Warren L

2017-05-11

Millions of patients worldwide require bone grafts for treatment of large, critically sized bone defects from conditions such as trauma, cancer, and congenital defects. Tissue engineered (TE) bone grafts have the potential to provide a more effective treatment than current bone grafts since they would restore fully functional bone tissue in large defects. Most bone TE approaches involve a combination of stem cells with porous, biodegradable scaffolds that provide mechanical support and degrade gradually as bone tissue is regenerated by stem cells. 3D-printing is a key technique in bone TE that can be used to fabricate functionalized scaffolds with patient-specific geometry. Using 3D-printing, composite polycaprolactone (PCL) and decellularized bone matrix (DCB) scaffolds can be produced to have the desired mechanical properties, geometry, and osteoinductivity needed for a TE bone graft. This book chapter will describe the protocols for fabricating and characterizing 3D-printed PCL:DCB scaffolds. Moreover, procedures for culturing adipose-derived stem cells (ASCs) in these scaffolds in vitro will be described to demonstrate the osteoinductivity of the scaffolds.

Considerations and Some Practical Solutions to Overcome Nanoparticle Interference with LAL Assays and to Avoid Endotoxin Contamination in Nanoformulations.

PubMed

Neun, Barry W; Dobrovolskaia, Marina A

2018-01-01

Monitoring endotoxin contamination in drugs and medical devices is required to avoid pyrogenic response and septic shock in patients receiving these products. Endotoxin contamination of engineered nanomaterials and nanotechnology-based medical products represents a significant translational hurdle. Nanoparticles often interfere with an in vitro Limulus Amebocyte Lysate (LAL) assay commonly used in the pharmaceutical industry for the detection and quantification of endotoxin. Such interference challenges the preclinical development of nanotechnology-formulated drugs and medical devices containing engineered nanomaterials. Protocols for analysis of nanoparticles using LAL assays have been reported before. Here, we discuss considerations for selecting an LAL format and describe a few experimental approaches for overcoming nanoparticle interference with the LAL assays to obtain more accurate estimation of endotoxin contamination in nanotechnology-based products. The discussed approaches do not solve all types of nanoparticle interference with the LAL assays but could be used as a starting point to address the problem. This chapter also describes approaches to prevent endotoxin contamination in nanotechnology-formulated products.

DNA Free Energy Landscapes and RNA Nano-Self-Assembly Using Atomic Force Microscopy

NASA Astrophysics Data System (ADS)

Frey, Eric William

There is an important conceptual lesson which has long been appreciated by those who work in biophysics and related interdisciplinary fields. While the extraordinary behavior of biological matter is governed by its detailed atomic structure and random fluctuations, and is therefore difficult to predict, it can nevertheless be understood within simplified frameworks. Such frameworks model the system as consisting of only one or a few components, and model the behavior of the system as the occupation of a single state out of a small number of states available. The emerging widespread application of nanotechnology, such as atomic force microscopy (AFM), has expanded this understanding in eye-opening new levels of detail by enabling nano-scale control, measurement, and visualization of biological molecules. This thesis describes two independent projects, both of which illuminate this understanding using AFM, but which do so from very different perspectives. The organization of this thesis is as follows. Chapter 1 begins with an experimental background and introduction to AFM, and then describes our setup in both single-molecule manipulation and imaging modes. In Chapter 2, we describe the first project, the motivation for which is to extend methods for the experimental determination of the free energy landscape of a molecule. This chapter relies on the analysis of single-molecule manipulation data. Chapter 3 describes the second project, the motivation for which is to create RNA-based nano-structures suitable for future applications in living mammalian cells. This chapter relies mainly on imaging. Chapters 2 and 3 can thus be read and understood separately.

Chapter 1: Creating Opportunities for Success through Replacement Services. Chapter 1 Replacement Class Collaborative Teaming and Intensive Teaching Time. A Model for Student Success in Newark City Schools.

ERIC Educational Resources Information Center

Stout, Kathleen

This collection of documents describes the Chapter 1 programs of the Newark (Ohio) City Schools and presents a model for programs to increase student success. Newark is a midsized city district with 1 high school, 3 middle schools, and 12 elementary schools, 7 of which receive Chapter 1 services. Collaborative efforts include replacement classes.…

Stimulus-Response Compatibility in Spatial Precuing and Symbolic Identification: Effects of Coding Practice, Retention and Transfer

DTIC Science & Technology

1989-05-31

for the effects. Most of the pqychophysiological research has examined event-related potentials (ERPs). Bashore (Chapter 7 ), Ragot (Chapter 8), and...flanking noise letters I signaled a response that was incongruent with the resTcnse i:.dicated b., a target letter. Bashore (Chapter 7 ) describes a...forms. First, compatibility effects have been used as estimates of interhemispheric transmission time (Bashore, I Chapter 7 ). The logic, articulated

Hanford Site National Environmental Policy Act (NEPA) Characterization

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

Neitzel, Duane A.; Antonio, Ernest J.; Eschbach, Tara O.

2001-09-01

This document describes the U.S. Department of Energy's (DOE) Hanford Site environment. It is updated each year and is intended to provide a consistent description of the Hanford Site environment for the many National Environmental Policy Act (NEPA) documents being prepared by DOE contractors. No statements of significance or environmental consequences are provided. This year's report is the thirteenth revision of the original document published in 1988 and is (until replaced by the fourteenth revision) the only version that is relevant for use in the preparation of Hanford NEPA, State Environmental Policy Act (SEPA), and Comprehensive Environmental Response, Compensation, andmore » Liability Act (CERCLA) documents. The two chapters included in this document (Chapters 4 and 6) are numbered to correspond to the chapters where such information is typically presented in environmental impact statements (Weiss) and other Hanford Site NEPA or CERCLA documentation. Chapter 4.0 (Affected Environment) describes Hanford Site climate and meteorology, geology, hydrology, ecology, cultural, archaeological, and historical resources, socioeconomics, occupational safety, and noise. Chapter 6.0 (Statutory and Regulatory Requirements) describes federal and state laws and regulations, DOE directives and permits, and presidential executive orders that are applicable to the NEPA documents prepared for Hanford Site activities.« less

Hanford Site National Environmental Policy Act (NEPA) Characterization

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

Neitzel, Duane A.; Bunn, Amoret L.; Duncan, Joanne P.

2002-09-01

This document describes the U.S. Department of Energy's (DOE) Hanford Site environment. It is updated each year and is intended to provide a consistent description of the Hanford Site environment for the many National Environmental Policy Act (NEPA) documents being prepared by DOE contractors. No statements of significance or environmental consequences are provided. This year's report is the thirteenth revision of the original document published in 1988 and is (until replaced by the fourteenth revision) the only version that is relevant for use in the preparation of Hanford NEPA, State Environmental Policy Act (SEPA), and Comprehensive Environmental Response, Compensation, andmore » Liability Act (CERCLA) documents. The two chapters included in this document (Chapters 4 and 6) are numbered to correspond to the chapters where such information is typically presented in environmental impact statements (Weiss) and other Hanford Site NEPA or CERCLA documentation. Chapter 4.0 (Affected Environment) describes Hanford Site climate and meteorology, geology, hydrology, ecology, cultural, archaeological, and historical resources, socioeconomics, occupational safety, and noise. Chapter 6.0 (Statutory and Regulatory Requirements) describes federal and state laws and regulations, DOE directives and permits, and presidential executive orders that are applicable to the NEPA documents prepared for Hanford Site activities.« less

Hanford Site National Environmental Policy Act (NEPA) Characterization, Revision 15

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

Neitzel, Duane A.; Bunn, Amoret L.; Burk, Kenneth W.

2003-09-01

This document describes the U.S. Department of Energy's (DOE) Hanford Site environment. It is updated each year and is intended to provide a consistent description of the Hanford Site environment for the many National Environmental Policy Act (NEPA) documents being prepared by DOE contractors. No statements of significance or environmental consequences are provided. This year's report is the thirteenth revision of the original document published in 1988 and is (until replaced by the fourteenth revision) the only version that is relevant for use in the preparation of Hanford NEPA, State Environmental Policy Act (SEPA), and Comprehensive Environmental Response, Compensation, andmore » Liability Act (CERCLA) documents. The two chapters included in this document (Chapters 4 and 6) are numbered to correspond to the chapters where such information is typically presented in environmental impact statements (Weiss) and other Hanford Site NEPA or CERCLA documentation. Chapter 4.0 (Affected Environment) describes Hanford Site climate and meteorology, geology, hydrology, ecology, cultural, archaeological, and historical resources, socioeconomics, occupational safety, and noise. Chapter 6.0 (Statutory and Regulatory Requirements) describes federal and state laws and regulations, DOE directives and permits, and presidential executive orders that are applicable to the NEPA documents prepared for Hanford Site activities.« less

Excited State Structural Dynamics of Carotenoids and ChargeTransfer Systems

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

Van Tassle, Aaron Justin

This dissertation describes the development andimplementation of a visible/near infrared pump/mid-infrared probeapparatus. Chapter 1 describes the background and motivation ofinvestigating optically induced structural dynamics, paying specificattention to solvation and the excitation selection rules of highlysymmetric molecules such as carotenoids. Chapter 2 describes thedevelopment and construction of the experimental apparatus usedthroughout the remainder of this dissertation. Chapter 3 will discuss theinvestigation of DCM, a laser dye with a fluorescence signal resultingfrom a charge transfer state. By studying the dynamics of DCM and of itsmethyl deuterated isotopomer (an otherwise identical molecule), we areable to investigate the origins of the charge transfer statemore » and provideevidence that it is of the controversial twisted intramolecular (TICT)type. Chapter 4 introduces the use of two-photon excitation to the S1state, combined with one-photon excitation to the S2 state of thecarotenoid beta-apo-8'-carotenal. These 2 investigations show evidencefor the formation of solitons, previously unobserved in molecular systemsand found only in conducting polymers Chapter 5 presents an investigationof the excited state dynamics of peridinin, the carotenoid responsiblefor the light harvesting of dinoflagellates. This investigation allowsfor a more detailed understanding of the importance of structuraldynamics of carotenoids in light harvesting.« less

Implications of fire management on cultural resources [Chapter 9

Treesearch

Rebecca S. Timmons; Leonard deBano; Kevin C. Ryan

2012-01-01

Previous chapters in this synthesis have identified the important fuel, weather, and fire relationships associated with damage to cultural resources (CR). They have also identified the types of effects commonly encountered in various fire situations and provided some guidance on how to recognize damages and minimize their occurrence. This chapter describes planning...

The Chief Student Affairs Officer: Leader, Manager, Mediator, Educator.

ERIC Educational Resources Information Center

Sandeen, Arthur

This book examines the leadership role of higher education chief student affairs officers (CSAO). Organized in three parts, Part One consists of four chapters that survey the CSAO's responsibilities as a campus leader. The first chapter describes the various roles and functions of the position and discusses its origins and evolution. Chapter Two…

Money and Education: A Guide to Illinois School Finance.

ERIC Educational Resources Information Center

McMaster, Donald; Sinkin, Judy G.

Illinois' education finance plan is described in the first of this report's two chapters, and the second chapter considers the finance plan's equity. Chapter 1 covers the state's Resource Equalizer Aid Program and the tax revenue it guarantees districts; the calculation of maximum tax guarantees and local shares; the apportionment of state aid;…

Federal Standards for Child Abuse and Neglect Prevention and Treatment Programs and Projects.

ERIC Educational Resources Information Center

Higgs, A. Catherine

Presented are federal standards designed to synthesize and describe the knowledge available on the prevention and treatment of child abuse and negect. A summary chapter (Chapter I) covers background information, organization and content of the standards, and utilization of the standards. Chapter II discusses the relationships among children,…

Pension Policy for a Mobile Labor Force.

ERIC Educational Resources Information Center

Turner, John A.; And Others

This book analyzes what happens to the pension benefits of workers who quit or are laid off jobs. The first chapter reviews the connection between job mobility and pension portability. Chapter 2 portrays a labor market undergoing changes that often result in reductions in retirement benefits. Chapter 3 describes job change further by examining…

Leading a Friends Helping Friends Peer Program.

ERIC Educational Resources Information Center

Painter, Carol

This manual is a guide for the adult learner who is developing and maintaining a peer counselor program. The first chapter presents an overview of peer counseling. The second chapter describes a model for a high school peer counseling program. Training, placements and programs, and a typical week's schedule are included. The third chapter presents…

Consumer Advertising: Its Role in Bringing a Product to Market. Revised Edition.

ERIC Educational Resources Information Center

Procter and Gamble Educational Services, Cincinnati, OH.

This kit, designed for high school classes, considers advertising from both consumers' and manufacturers' perspectives. The role of advertising in relation to free enterprise principles is discussed in chapter 1, while chapter 2 provides a history of U.S. advertising processes and development. Chapter 3 describes advertising's role in bringing a…

Historical Beginnings...The Federal Reserve.

ERIC Educational Resources Information Center

Johnson, Roger T.

The booklet presents an historical narrative of banks and banking events leading to the establishment of the Federal Reserve System in 1914. The document is divided into three chapters. Chapter I discusses attempts to establish a bank before the 20th century, describing the creation of two central banks whose chapters were allowed to lapse, the…

Developmental Education: Preparing Successful College Students. Monograph Series #24.

ERIC Educational Resources Information Center

Higbee, Jeanne L., Ed.; Dwinell, Patricia L., Ed.

The 16 chapters of this volume describe a wide variety of developmental programs intended to promote skill development and enhance academic performance for high-risk students at all levels of higher education. Following an introductory chapter by the editors, the chapters are: (1) "The Origin, Scope, and Outcomes of Developmental Education in the…

A Story About You.

ERIC Educational Resources Information Center

Southard, Helen

This pamphlet describes the process of human growth from conception to adolescence and is designed for use in grades four through six. Chapter One discusses growth and Chapter Two discusses the beginning of life. Chapter Three is concerned with growth from conception on to birth. The birth of the babies, both human and other mammals, and helping…

Micropropagation of Iris sp.

PubMed

Jevremović, Slađana; Jeknić, Zoran; Subotić, Angelina

2013-01-01

Irises are perennial plants widely used as ornamental garden plants or cut flowers. Some species accumulate secondary metabolites, making them highly valuable to the pharmaceutical and perfume industries. Micropropagation of irises has successfully been accomplished by culturing zygotic embryos, different flower parts, and leaf base tissues as starting explants. Plantlets are regenerated via somatic embryogenesis, organogenesis, or both processes at the same time depending on media composition and plant species. A large number of uniform plants are produced by somatic embryogenesis, however, some species have decreased morphogenetic potential overtime. Shoot cultures obtained by organogenesis can be multiplied for many years. Somatic embryogenic tissue can be reestablished from leaf bases of in vitro-grown shoots. The highest number of plants can be obtained by cell suspension cultures. This chapter describes effective in vitro plant regeneration protocols for Iris species from different types of explants by somatic embryogenesis and/or organogenesis suitable for the mass propagation of ornamental and pharmaceutical irises.

Analysis of DNA methylation in FFPE tissues using the MethyLight technology.

PubMed

Dallol, Ashraf; Al-Ali, Waleed; Al-Shaibani, Amina; Al-Mulla, Fahd

2011-01-01

Novel biomarkers are sought after by mining DNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues. Such tissues offer the great advantage of often having complete clinical data (including survival), as well as the tissues are amenable for laser microdissection targeting specific tissue areas. Downstream analysis of such DNA includes mutational screens and methylation profiling. Screening for mutations by sequencing requires a significant amount of DNA for PCR and cycle sequencing. This is self-inhibitory if the gene screened has a large number of exons. Profiling DNA methylation using the MethyLight technology circumvents this problem and allows for the mining of several biomarkers from DNA extracted from a single microscope slide of the tissue of interest. We describe in this chapter a detailed protocol for MethyLight and its use in the determination of CpG Island Methylator Phenotype status in FFPE colorectal cancer samples.

Measuring mitochondrial uncoupling protein-2 level and activity in insulinoma cells.

PubMed

Barlow, Jonathan; Hirschberg, Verena; Brand, Martin D; Affourtit, Charles

2013-01-01

Mitochondrial uncoupling protein-2 (UCP2) regulates glucose-stimulated insulin secretion (GSIS) by pancreatic beta cells-the physiological role of the beta cell UCP2 remains a subject of debate. Experimental studies informing this debate benefit from reliable measurements of UCP2 protein level and activity. In this chapter, we describe how UCP2 protein can be detected in INS-1 insulinoma cells and how it can be knocked down by RNA interference. We demonstrate briefly that UCP2 knockdown lowers glucose-induced rises in mitochondrial respiratory activity, coupling efficiency of oxidative phosphorylation, levels of mitochondrial reactive oxygen species, and insulin secretion. We provide protocols for the detection of the respective UCP2 phenotypes, which are indirect, but invaluable measures of UCP2 activity. We also introduce a convenient method to normalize cellular respiration to cell density allowing measurement of UCP2 effects on specific mitochondrial oxygen consumption. Copyright © 2013 Elsevier Inc. All rights reserved.

Plastid transformation in cabbage (Brassica oleracea L. var. capitata L.) by the biolistic process.

PubMed

Tseng, Menq-Jiau; Yang, Ming-Te; Chu, Wan-Ru; Liu, Cheng-Wei

2014-01-01

Cabbage (Brassica oleracea L. var. capitata L.) is one of the most important vegetable crops grown worldwide. Scientists are using biotechnology in addition to traditional breeding methods to develop new cabbage varieties with desirable traits. Recent biotechnological advances in chloroplast transformation technology have opened new avenues for crop improvement. In 2007, we developed a stable plastid transformation system for cabbage and reported the successful transformation of the cry1Ab gene into the cabbage chloroplast genome. This chapter describes the methods for cabbage transformation using biolistic procedures. The following sections are included in this protocol: preparation of donor materials, coating gold particles with DNA, biolistic bombardment, as well as the regeneration and selection of transplastomic cabbage plants. The establishment of a plastid transformation system for cabbage offers new possibilities for introducing new agronomic and horticultural traits into Brassica crops.

Screening and Evaluation of Medications for Treating Cannabis Use Disorder

PubMed Central

Panlilio, Leigh V.; Justinova, Zuzana; Trigo, Jose M.; Le Foll, Bernard

2016-01-01

Cannabis use has been increasingly accepted legally and in public opinion. However, cannabis has the potential to produce adverse physical and mental health effects and can result in cannabis use disorder (CUD) in a substantial percentage of both occasional and daily cannabis users. Many people have difficulty discontinuing use. Therefore, it would be beneficial to develop safe and effective medications for treating CUD. To achieve this, methods have been developed for screening and evaluating potential medications using animal models and controlled experimental protocols in human volunteers. In this chapter we describe: 1) animal models available for assessing the effect of potential medications on specific aspects of CUD; 2) the main findings obtained so far with these animal models; 3) the approaches used to assess potential medications in humans in laboratory experiments and clinical trials; and 4) the effectiveness of several potential pharmacotherapies on the particular aspects of CUD modeled in these human studies. PMID:27055612

The Stem Cell Laboratory: Design, Equipment, and Oversight

PubMed Central

Wesselschmidt, Robin L.; Schwartz, Philip H.

2013-01-01

This chapter describes some of the major issues to be considered when setting up a laboratory for the culture of human pluripotent stem cells (hPSCs). The process of establishing a hPSC laboratory can be divided into two equally important parts. One is completely administrative and includes developing protocols, seeking approval, and establishing reporting processes and documentation. The other part of establishing a hPSC laboratory involves the physical plant and includes design, equipment and personnel. Proper planning of laboratory operations and proper design of the physical layout of the stem cell laboratory so that meets the scope of planned operations is a major undertaking, but the time spent upfront will pay long-term returns in operational efficiency and effectiveness. A well-planned, organized, and properly equipped laboratory supports research activities by increasing efficiency and reducing lost time and wasted resources. PMID:21822863

RNA-Seq-Based Transcript Structure Analysis with TrBorderExt.

PubMed

Wang, Yejun; Sun, Ming-An; White, Aaron P

2018-01-01

RNA-Seq has become a routine strategy for genome-wide gene expression comparisons in bacteria. Despite lower resolution in transcript border parsing compared with dRNA-Seq, TSS-EMOTE, Cappable-seq, Term-seq, and others, directional RNA-Seq still illustrates its advantages: low cost, quantification and transcript border analysis with a medium resolution (±10-20 nt). To facilitate mining of directional RNA-Seq datasets especially with respect to transcript structure analysis, we developed a tool, TrBorderExt, which can parse transcript start sites and termination sites accurately in bacteria. A detailed protocol is described in this chapter for how to use the software package step by step to identify bacterial transcript borders from raw RNA-Seq data. The package was developed with Perl and R programming languages, and is accessible freely through the website: http://www.szu-bioinf.org/TrBorderExt .

Construction of a filamentous phage display peptide library.

PubMed

Fagerlund, Annette; Myrset, Astrid Hilde; Kulseth, Mari Ann

2014-01-01

The concept of phage display is based on insertion of random oligonucleotides at an appropriate location within a structural gene of a bacteriophage. The resulting phage will constitute a library of random peptides displayed on the surface of the bacteriophages, with the encoding genotype packaged within each phage particle. Using a phagemid/helper phage system, the random peptides are interspersed between wild-type coat proteins. Libraries of phage-expressed peptides may be used to search for novel peptide ligands to target proteins. The success of finding a peptide with a desired property in a given library is highly dependent on the diversity and quality of the library. The protocols in this chapter describe the construction of a high-diversity library of phagemid vector encoding fusions of the phage coat protein pVIII with random peptides, from which a phage library displaying random peptides can be prepared.

Synchronization of Somatic Embryogenesis in Date Palm Suspension Culture Using Abscisic Acid.

PubMed

Alwael, Hussain A; Naik, Poornananda M; Al-Khayri, Jameel M

2017-01-01

Somatic embryogenesis is considered the most effective method for commercial propagation of date palm. However, the limitation of obtaining synchronized development of somatic embryos remains an impediment. The synchronization of somatic embryo development is ideal for the applications to produce artificial seeds. Abscisic acid (ABA) is associated with stress response and influences in vitro growth and development. This chapter describes an effective method to achieve synchronized development of somatic embryos in date palm cell suspension culture. Among the ABA concentrations tested (0, 1, 10, 50, 100 μM), the best synchronized growth was obtained in response to 50-100 μM. Here we provide a comprehensive protocol for in vitro plant regeneration of date palm starting with shoot-tip explant, callus initiation and growth, cell suspension establishment, embryogenesis synchronization with ABA treatment, somatic embryo germination, and rooting as well as acclimatized plantlet establishment.

Forest management practices and silviculture. Chapter 12.

Treesearch

Donald A. Perala; Elon S. Verry

2011-01-01

This chapter is an overview of forest management and silviculture practices, and lessons learned, on the Marcell Experimental Forest (MEF). The forests there are a mosaic of natural regeneration and conifer plantations. Verry (1969) described forest-plant communities in detail for the study watersheds (Sl through S6) on the MEF. The remaining area is described in...

Technical Report and Data File User's Manual for the 1992 National Adult Literacy Survey.

ERIC Educational Resources Information Center

Kirsch, Irwin; Yamamoto, Kentaro; Norris, Norma; Rock, Donald; Jungeblut, Ann; O'Reilly, Patricia; Berlin, Martha; Mohadjer, Leyla; Waksberg, Joseph; Goksel, Huseyin; Burke, John; Rieger, Susan; Green, James; Klein, Merle; Campbell, Anne; Jenkins, Lynn; Kolstad, Andrew; Mosenthal, Peter; Baldi, Stephane

Chapter 1 of this report and user's manual describes design and implementation of the 1992 National Adult Literacy Survey (NALS). Chapter 2 reviews stages of sampling for national and state survey components; presents weighted and unweighted response rates for the household component; and describes non-incentive and prison sample designs. Chapter…

Drawing from the Well. Oral History and Folk Arts in the Classroom and Community.

ERIC Educational Resources Information Center

Silnutzer, Randi, Ed.; Watrous, Beth Eildin, Ed.

Each chapter of this document describes a different project and approach for introducing students (elementary to high school) to oral history and folk arts. All chapters use a standard format in which a general overview of the project, describing themes, philosophies, and methods are followed by sample lesson plans, teacher guidelines, and student…

The Family Transition Program: Implementation and Three-Year Impacts of Florida's Initial Time-Limited Welfare Program.

ERIC Educational Resources Information Center

Bloom, Dan; Farrell, Mary; Kemple, James J.; Verma, Nandita

This report focuses on implementation, impacts, and time limit in Florida's Family Transition Program (FTP), a welfare reform initiative. Chapter 1 describes FTP and evaluation, target population, and data. Chapter 2 describes implementation of FTP in Escambia County: staffing and organizational structure of FTP and traditional Aid to Families…

National Job Corps Study: Report on the Process Analysis. Research and Evaluation Report Series.

ERIC Educational Resources Information Center

Johnson, Terry; Gritz, Mark; Jackson, Russell; Burghardt, John; Boussy, Carol; Leonard, Jan; Orians, Carlyn

This report presents results of a process analysis that describes and documents Job Corps services and operations. Chapter one provides overviews of Job Corps, the national Job Corps study, and the process analysis. Chapter two describes the administrative structure of Job Corps and presents data on the geographic distribution and characteristics…

In First Person Plural: Growing Up with a Disadvantaged Community.

ERIC Educational Resources Information Center

Salach, Simcha

From the perspective of the project leader, this book traces the growth and development of the Early Childhood and Family Education Project in Morasha, Israel, during the 8 years of its existence (1982-1990). Chapter 1 describes Morasha, a poor, immigrant neighborhood in Ramat HaSharon, and the author's childhood there. Chapter 2 describes the…

An Evaluation of the Self-Service Approach in One-Stop Career Centers. Final Report.

ERIC Educational Resources Information Center

D'Amico, Ronald; Fedrau, Ruth; Kimball, Mary; Midling, Michael; Soukamneuth, Sengsouvanh

This report presents findings from a study of self-service systems in eight One-Stop Career Centers that were identified as being particularly efficacious or noteworthy for completeness of resources, ease of use, or outreach to special populations. Chapter I describes the study design. Chapter II describes physical facilities. It focuses on the…

Establishing a Group Counseling Program for Elementary School Children Who Have Experienced Parental Divorce.

ERIC Educational Resources Information Center

Goldstein, Alan

This practicum report describes a counseling program established in the school setting for young children who have experienced parental divorce. Chapter I describes the Merrytown Energyville Regional School System, New Jersey, in which the program was established and notes the author's role and responsibilities. Chapter II presents the results of…

A Volcano Rekindled: The Renewed Eruption of Mount St. Helens, 2004-2006

USGS Publications Warehouse

Sherrod, David R.; Scott, William E.; Stauffer, Peter H.

2008-01-01

Mount St. Helens began a dome-building eruption in September 2004 after nearly two decades of quiescence. Dome growth was initially robust, became more sluggish with time, and ceased completely in late January 2008. The volcano has been quiet again since January 2008. Professional Paper 1750 describes the first 1 1/2 years of this eruptive activity, chiefly from September 2004 until December 2005. Its 37 chapters contain contributions of 87 authors from 23 institutions, including the U.S. Geological Survey, Forest Service, many universities, and local and State emergency management agencies. Chapter topics range widely - from seismology, geology, geodesy, gas geochemistry, and petrology to the human endeavor required for managing the public volcanic lands and distributing information during the hectic early days of a renewed eruption. In PDF format, the book may be downloaded in its entirety or by its topical sections, each section including a few prefatory paragraphs that describe the general findings, recurrent themes, and, in some cases, the unanswered questions that arise repeatedly. Those readers who prefer downloading the smaller files of only a chapter or two have this option available as well. Readers are directed to chapter 1 for a general overview of the eruption and the manner in which different chapters build our knowledge of events. More detailed summaries for specific topics can be found in chapter 2 (seismology), chapter 9 (geology), chapter 14 (deformation), chapter 26 (gas geochemistry), and chapter 30 (petrology). The printed version of the book may be purchased as a hardback weighty tome (856 printed pages) that includes a DVD replete with the complete online version, including all chapters and several additional appendixes not in the printed book.

Preliminary Understanding of Surface Plasmon-Enhanced Circular Dichroism Spectroscopy by Single Particle Imaging

NASA Astrophysics Data System (ADS)

Zhan, Kangshu

Monitoring chiral optical signals of biomolecules as their conformation changes is an important means to study their structures, properties, and functions. Most measurements, however, are ensemble measurements because chiral optical signals from a single biomolecule is often too weak to be detected. In this dissertation, I present my early attempts to study conformational changes of adsorbed proteins by taking advantage of the enhanced electromagnetic (EM) field around a well-designed plasmonic nanofeature. In particular, I discuss the detection of protein adsorption and denaturation on metallic nanoparticles using single particle scattering and CD spectroscopic imaging. Particles of two distinctively different sizes were compared and two different sample protein molecules were studied. A combination of experimental and computational tools was used to simulate and interpret the collected scattering and CD results. The first chapter provides a brief overview of the state-of-art research in CD spectroscopic studies at the single particle level. Three different means to make particles capable of chiral detection are discussed. Various applications beyond single particle imaging are presented to showcase the potential of the described research project, beyond our immediate goals. The second chapter describes my initial characterization of large, metallic, anisotropic nanorods and the establishment of experimental procedures used later for spectrum reconstruction, data visualization and analysis. The physical shape and structure of the particles were imaged by scanning electron microscopy (SEM), the chemical composition by energy dispersive X-ray Spectroscopy (EDS), and the optical properties by darkfield microscopy. An experimental protocol was developed to connect information collected from separate techniques for the same particle, with the aims of discovering any possible structural-property correlation. The reproducibility of the single particle imaging method was evaluated. Full spectrum reconstruction using a set of selected optical filters was carried out and data visualization using a Matlab based 3D mapping method was demonstrated. The third chapter describes the introduction of biomolecules in chiral particle studies. By measuring the circular dichroism spectrum and image of nanorods during lysozyme adsorption and denaturation, I was able to monitor the conformation change of proteins on large gapped nanorods. Experiment results suggested that the conformational change of absorbed protein could lead to the change of chiral signal of nanoparticles, suggesting the potentials of detecting biomolecular structural changes at the single nanoparticle level, though much uncertainty still present. The inherent high background of large, gapped nanoparticles when they interact with biomolecules led to the research described in the 4th chapter where I studied small palladium-silver coreshell nanoparticle properties and its interaction with proteins. SEM was used to characterize particles structures; UV-Vis and darkfield microscopy was used to capture particles' optical responses; and the finite-difference time-domain (FDTD) method was used to simulate resulting spectra and to compare with experimental outcomes. Lysozyme and bovine serum albumin (BSA) were used as the model molecules to study their conformational changes after being adsorbed onto particles. Last but the least, the 5th chapter is dedicated to FDTD simulation of a pair of perfectly shaped triangle nanoprisms to illustrate possible CD responses to be expected from extreme particles with sharp corners and much concentrated local EM field. Different coupling modes of triangle nanoprism were analyzed. It is found that many factors, such as particle orientation, spacing, and their relative position, could lead to significantly different coupling efficient, for both homodimers and heterodimers. The modeling data suggested interesting potentials of nanoparticles of extreme geometric features for high sensitivity surface plasmon-enhanced CD imaging at the signal particle level.

By-Laws | Center for Cancer Research

Cancer.gov

The NIH Fellows Editorial Board (FEB) offers a free and confidential editorial service for NIH fellows. The Board members edit manuscripts, abstracts, book chapters, clinical protocols, and grant submissions and offer suggestions for grammar, structure, and style, but not for scientific content. The Board provides valuable training and editorial experience to its members for academic as well as nonacademic scientific careers.

Preliminary Phytochemical Studies.

PubMed

2016-01-01

Plants are the natural producers of medicinal agents like alkaloids, flavonoids, tannins, and phenolics. These phytocompounds alone or in combination act as a therapeutic agent in various disease complications. Various chemical reagents are used to determine the major phytochemicals present in plant parts. Protocols involved in screening of alkaloids, carbohydrates, glycosides, saponins, phytosterols, fixed oils, and fats are shown in this chapter.

Postsuicide Intervention as a Prevention Tool: Developing a Comprehensive Campus Response to Suicide and Related Risk

ERIC Educational Resources Information Center

Cimini, M. Dolores; Rivero, Estela M.

2013-01-01

This chapter explores the critical role of crisis intervention and other support after a suicide has occurred as part of a comprehensive suicide prevention response within college and university campuses. The important components of postsuicide intervention campus crisis response and protocols and the identification of key stakeholders to…

When Children Are Abused: An Educator's Guide to Intervention.

ERIC Educational Resources Information Center

Crosson-Tower, Cynthia

This book presents guidance for educators on recognizing and responding to different forms of child maltreatment as well as ideas on the formation of a school reporting protocol and a Child Protection Team. The 11 chapters are: (1) "Why Are Educators So Important in the Lives of Abused and Neglected Children?"; (2) "How Can We…

Metamaterial Resonant Absorbers for Terahertz Sensing

DTIC Science & Technology

2015-12-01

Equation Chapter 1 Section 1 NAVAL POSTGRADUATE SCHOOL MONTEREY, CALIFORNIA THESIS Approved for public release; distribution...Co-Advisor Fabio Alves THIS PAGE INTENTIONALLY LEFT BLANK i REPORT DOCUMENTATION PAGE Form Approved OMB No. 0704–0188 Public reporting...Department of Defense or the U.S. government. IRB Protocol number ____N/A____. 12a. DISTRIBUTION / AVAILABILITY STATEMENT Approved for public

Living Together in Space: The Design and Operation of the Life Support Systems on the International Space Station. Volume 1

NASA Technical Reports Server (NTRS)

Wieland, P. O.

1998-01-01

The International Space Station (ISS) incorporates elements designed and developed by an international consortium led by the United States (U.S.), and by Russia. For this cooperative effort to succeed, it is crucial that the designs and methods of design of the other partners are understood sufficiently to ensure compatibility. Environmental Control and Life Support (ECLS) is one system in which functions are performed independently on the Russian Segment (RS) and on the U.S./international segments. This document describes, in two volumes, the design and operation of the ECLS Systems (ECLSS) on board the ISS. This current volume, Volume 1, is divided into three chapters. Chapter 1 is a general overview of the ISS, describing the configuration, general requirements, and distribution of systems as related to the ECLSS, and includes discussion of the design philosophies of the partners and methods of verification of equipment. Chapter 2 describes the U.S. ECLSS and technologies in greater detail. Chapter 3 describes the ECLSS in the European Attached Pressurized Module (APM), Japanese Experiment Module (JEM), and Italian Mini-Pressurized Logistics Module (MPLM). Volume II describes the Russian ECLSS and technologies in greater detail. These documents present thorough, yet concise, descriptions of the ISS ECLSS.

The Art of Investing School District Funds: Rules of the Game.

ERIC Educational Resources Information Center

Flynn, Jeffrey

The "art" of investment and the relationships between the cash manager and the outside professionals who implement the steps that entail cash management are the subjects of the 11 chapters of this guide to investing school district funds. Chapter 1 describes the role and responsibility of the small school district cash manager. Chapter 2 discusses…

Managing Asbestos in Place: A Building Owner's Guide to Operations and Maintenance Programs for Asbestos-Containing Materials.

ERIC Educational Resources Information Center

Environmental Protection Agency, Washington, DC.

Instructions for building owners on the selection and application of appropriate asbestos control and abatement actions are presented in this guidebook. Chapter 1 offers background information on the asbestos problem. Chapter 2 describes the purpose and scope of an operations and maintenance (O&M) program. The third chapter discusses planning…

Calculus, Part 3, Teacher's Commentary, Unit No. 71. Revised Edition.

ERIC Educational Resources Information Center

Beck, A.; And Others

This is part three of a three-part manual for teachers using SMSG high school text materials. The overall purpose for each of the chapters is described and the mathematical development detailed. Background information for key concepts and answers for all exercises in each chapter are provided. Chapter topics include: (1) vectors and curves; (2)…

Planning a Successful Conference. Survival Skills for Scholars, Volume 13.

ERIC Educational Resources Information Center

Winter, Cynthia

This book is designed to guide a university faculty member through each stage of conference planning. Each chapter in the book describes a step in the meeting planning process. The first chapter is devoted to learning about professional conferences and meetings from an attendee's perspective. Other chapters address: designing a meeting (who, what,…

Adlerian Counseling: A Practical Approach for a New Decade. Third Edition.

ERIC Educational Resources Information Center

Sweeney, Thomas J.

This book describes the practical applications of the psychology of Alfred Adler. The first chapter provides a short introduction to Adler and his work. Chapter 2 contains a compendium of the latest research and practices on the characteristics of healthy persons as they relate to individual psychology. Chapters 3 and 4 explain dimensions and uses…

Guide to Non-Sexist Early Childhood Education.

ERIC Educational Resources Information Center

Sprung, Barbara

This guide describes the development of a nonsexist early childhood curriculum designed to make teachers and parents more aware of the sexism existing in society and reflected in our schools. The guide is divided into six chapters: Chapter 1 presents a brief history of the Non-Sexist Child Development Project; Chapter 2 examines some of the basic…

Ultracool Dwarfs and their companions

NASA Astrophysics Data System (ADS)

Blake, Cullen H.

This thesis explores new techniques for making precise measurements of low-mass stars and brown dwarfs, collectively known as Ultracool Dwarfs (UCDs). These new techniques are directly applicable to the search for extrasolar planets and efforts to test theoretical models of stellar structure and evolution at the bottom of the main sequence. The first three chapters of this thesis describe the development and application of a new technique for making radial velocity measurements of UCDs at near infrared (NIR) wavelengths. The first chapter describes a pilot study that demonstrates a significant improvement over previous work on Doppler measurements in the NIR. Using this technique we have carried out a Doppler survey of 65 L dwarfs. The second chapter describes the discovery of a new spectroscopic binary that may be one of the most important for constraining theoretical models of UCDs. The third chapter describes the Doppler survey in detail and presents measurements of a new spectroscopic binary system that is an excellent candidate for a giant planetary companion to a mid-L dwarf. This chapter also includes a discussion of the of the rotation, space motions, and binarity of the L dwarfs in the survey sample. The fourth chapter describes efforts to obtain precise photometric measurements of UCDs with the Peters Automated Infrared Imaging Telescope (PAIRITEL). Using software scheduling and data reduction systems designed in part by the author, PAIRITEL gathered more than 10 6 seconds of observations of a sample of 20 UCDs. We investigate the limitations to ground-based infrared photometry and characterize the ability of a system like PAIRITEL to detect transits of UCDs by Earth-like planets. The fifth chapter explores the potential impact of future synoptic surveys on studies of UCDs. Surveys like Pan-STARRS and LSST will obtain a small number of high-quality observations of a large number of UCDs. Using data from the Sloan Digital Sky Survey, we demonstrate that such data can be used to reliably detect low-mass eclipsing binary stars. We present the discovery of a double- lined eclipsing binary system that allows us to directly measure the masses and radii of two M dwarfs.

Regulated Expression of Adenoviral Vectors-Based Gene Therapies

PubMed Central

Curtin, James F.; Candolfi, Marianela; Puntel, Mariana; Xiong, Weidong; Muhammad, A. K. M.; Kroeger, Kurt; Mondkar, Sonali; Liu, Chunyan; Bondale, Niyati; Lowenstein, Pedro R.; Castro, Maria G.

2008-01-01

Summary Regulatable promoter systems allow gene expression to be tightly controlled in vivo. This is highly desirable for the development of safe, efficacious adenoviral vectors that can be used to treat human diseases in the clinic. Ideally, regulatable cassettes should have minimal gene expression in the “OFF” state, and expression should quickly reach therapeutic levels in the “ON” state. In addition, the components of regulatable cassettes should be non-toxic at physiological concentrations and should not be immunogenic, especially when treating chronic illness that requires long-lasting gene expression. In this chapter, we will describe in detail protocols to develop and validate first generation (Ad) and high-capacity adenoviral (HC-Ad) vectors that express therapeutic genes under the control of the TetON regulatable system. Our laboratory has successfully used these protocols to regulate the expression of marker genes, immune stimulatory genes, and toxins for cancer gene therapeutics, i.e., glioma that is a deadly form of brain cancer. We have shown that this third generation TetON regulatable system, incorporating a doxycycline (DOX)-sensitive rtTA2S-M2 inducer and tTSKid silencer, is non-toxic, relatively non-immunogenic, and can tightly regulate reporter transgene expression downstream of a TRE promoter from adenoviral vectors in vitro and also in vivo. PMID:18470649

Ion-exchange chromatography: basic principles and application to the partial purification of soluble mammalian prolyl oligopeptidase.

PubMed

Cummins, Philip M; Dowling, Oonagh; O'Connor, Brendan F

2011-01-01

Ion-exchange chromatography (IEC) allows for the separation of ionizable molecules on the basis of differences in charge properties. Its large sample-handling capacity, broad applicability (particularly to proteins and enzymes), moderate cost, powerful resolving ability, and ease of scale-up and automation have led to it becoming one of the most versatile and widely used of all liquid chromatography (LC) techniques. In this chapter, we review the basic principles of IEC, as well as the broader criteria for selecting IEC conditions. By way of further illustration, we outline protocols necessary to partially purify a serine peptidase from bovine whole brain cytosolic fraction, covering crude tissue extract preparation through to partial purification of the target enzyme using anion-exchange chromatography. Protocols for assaying total protein and enzyme activity in both pre- and post-IEC fractions are also described. The target serine peptidase, prolyl oligopeptidase (POP, EC3.4.21.26), is an 80-kDa enzyme with endopeptidase activity towards peptide substrates of ≤30 amino acids. POP is a ubiquitous post-proline cleaving enzyme with particularly high expression levels in the mammalian brain, where it participates in the metabolism of neuroactive peptides and peptide-like hormones (e.g. thyroliberin, gonadotropin-releasing hormone).

Investigation of Lung Structure-Function Relationships Using Hyperpolarized Noble Gases

NASA Astrophysics Data System (ADS)

Thomen, Robert P.

Magnetic Resonance Imaging (MRI) is an application of the nuclear magnetic resonance (NMR) phenomenon to non-invasively generate 3D tomographic images. MRI is an emerging modality for the lung, but it suffers from low sensitivity due to inherent low tissue density and short T(*/2) . Hyperpolarization is a process by which the nuclear contribution to NMR signal is greatly enhanced to more than 100,000 times that of samples in thermal equilibrium. The noble gases 3He and 129Xe are most often hyperpolarized by transfer of light angular momentum through the electron of a vaporized alkali metal to the noble gas nucleus (called Spin Exchange Optical Pumping). The enhancement in NMR signal is so great that the gas itself can be imaged via MRI, and because noble gases are chemically inert, they can be safely inhaled by a subject, and the gas distribution within the interior of the lung can be imaged. The mechanics of respiration is an elegant physical process by which air is is brought into the distal airspaces of the lungs for oxygen/carbon dioxide gas exchange with blood. Therefore proper description of lung function is intricately related to its physical structure , and the basic mechanical operation of healthy lungs -- from pressure driven airflow, to alveolar airspace gas kinetics, to gas exchange by blood/gas concentration gradients, to elastic contraction of parenchymal tissue -- is a process decidedly governed by the laws of physics. This dissertation will describe experiments investigating the relationship of lung structure and function using hyperpolarized (HP) noble gas MRI. In particular HP gases will be applied to the study of several pulmonary diseases each of which demonstrates unique structure-function abnormalities: asthma, cystic fibrosis, and chronic obstructive pulmonary disease. Successful implementation of an HP gas acquisition protocol for pulmonary studies is an involved and stratified undertaking which requires a solid theoretical foundation in NMR and hyperpolarization theory, construction of dedicated hardware, development of dedicated software, and appropriate image analysis techniques for all acquired data. The author has been actively involved in each of these and has dedicated specific chapters of this dissertation to their description. First, a brief description of lung structure-function investigations and pulmonary imaging will be given (chapter 1). Brief discussions of basic NMR, MRI, and hyperpolarization theory will be given (chapters 2 and 3) followed by their particular methods of implementation in this work (chapters 4 and 5). Analysis of acquired HP gas images will be discussed (chapter 6), and the investigational procedures and results for each lung disease examined will be detailed (chapter 7). Finally, a quick digression on the strengths and limitations of HP gas MRI will be provided (chapter 8).

Comment on Cross, Fine, Jones, and Walsh (2012): Good Therapeutic Services--Therapeutic Advocacy and Forensic Neutrality

ERIC Educational Resources Information Center

Connell, Mary

2012-01-01

Cross, Fine, Jones, and Walsh (2012) provided a thoughtful review and critique of a book chapter describing the interview process at Child Advocacy Centers. They observed some of the ways that concerns raised in that chapter are being addressed and described revised guidelines that further clarify issues. Ongoing research and examination of the…

Reactivity of lignin and lignans: Correlation with molecular orbital calculations

Treesearch

Thomas Elder

2010-01-01

To date, and as can be seen from the other chapters of this text, the structure and chemistry of lignin have been described in terms of results from a wide range of chemical or spectroscopic methods to construct a mosaic picture of the polymer. The current chapter continues this process by describing past, present and potential applications of electronic structure...

Regional geology and tectonics

USGS Publications Warehouse

Nokleberg, Warren J.; Bundtzen, Thomas K.; Nokleberg, Warren J.; Price, Raymond A.; Scholl, David W.; Stone, David B.

2017-01-01

This chapter describes the regional geology and tectonic origins of the major geologic units for the Northern Cordillera. The goals of the chapter are to: (1) provide a summary and regional overview of this vast region that contains a complicated geologic history; and (2) describe the major geologic units and tectonic events that cover a broad geologic time span from the Proterozoic to the Holocene (Recent).

Experimental Course Development in Introductory Economics at Indiana University. The Journal of Economic Education, Special Issue No. 4, Fall 1975.

ERIC Educational Resources Information Center

Saunders, Phillip

A two part experimental introductory college economics course is described. Data on the combination macroeconomics and microeconomics course have been collected over eight consecutive terms and are presented in nine chapters. Chapter I describes course goals as stimulation of student interest, teaching a few basic economic principles, helping…

Post-16 Provision in CTCs: Bridging the Divide. Number 6 in a Series of CTC Trust Publications.

ERIC Educational Resources Information Center

Jones, Ruth

This document is intended to describe the approach of the City Technology Colleges (CTCs) in Great Britain to provide young people older than 16 with a vocationally oriented curriculum while still ensuring their intellectual and cultural development. Following a summary, the first chapter is an introduction. The second chapter describes the old…

Facilities Information Management: A Guide for State and Local Education Agencies. NCES 2003-400

ERIC Educational Resources Information Center

National Center for Education Statistics (ED), Washington, DC.

This Guide has been developed to provide a framework for decision makers, school facility managers, and the public to identify a basic set of school facilities data elements, including definitions that will meet their information needs. Chapter 1 describes the purpose, scope, and intended audience for this guide. Chapter 2 describes how to use the…

Landsat and Thermal Infrared Imaging

NASA Technical Reports Server (NTRS)

Arvidson, Terry; Barsi, Julia; Jhabvala, Murzy; Reuter, Dennis

2012-01-01

The purpose of this chapter is to describe the collection of thermal images by Landsat sensors already on orbit and to introduce the new thermal sensor to be launched in 2013. The chapter describes the thematic mapper (TM) and enhanced thematic mapper plus (ETM+) sensors, the calibration of their thermal bands, and the design and prelaunch calibration of the new thermal infrared sensor (TIRS).

U 3Si 2 Fabrication and Testing for Implementation into the BISON Fuel Performance Code

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

Knight, Travis W.

A creep test stand was designed and constructed for compressive creep testing of U 3Si 2 pellets. This is described in Chapter 3. Creep testing of U 3Si 2 pellets was completed. In total, 13 compressive creep tests of U 3Si 2 pellets was successfully completed. This is reported in Chapter 3. Secondary creep model of U 3Si 2 was developed and implemented in BISON. This is described in Chapter 4. Properties of U 3Si 2 were implemented in BISON. This is described in Chapter 4. A resonant frequency and damping analyzer (RFDA) using impulse excitation technique (IET) was setup,more » tested, and used to analyze U 3Si 2 samples to measure Young’s and Shear Moduli which were then used to calculate the Poisson ratio for U 3Si 2. This is described in Chapter 5. Characterization of U 3Si 2 samples was completed. Samples were prepared and analyzed by XRD, SEM, and optical microscopy. Grain size analysis was conducted on images. SEM with EDS was used to analyze second phase precipitates. Impulse excitation technique was used to determine the Young’s and Shear Moduli of a tile specimen which allowed for the determination of the Poisson ratio. Helium pycnometry and mercury intrusion porosimetry was performed and used with image analysis to determine porosity size distribution. Vickers microindentation characterization method was used to evaluate the mechanical properties of U 3Si 2 including toughness, hardness, and Vickers hardness. Electrical resistivity measurement was done using the four-point probe method. This is reported in Chapter 5.« less

Fabrication and application of advanced functional materials from lignincellulosic biomass

NASA Astrophysics Data System (ADS)

Hu, Sixiao

This dissertation explored the conversion of lignocellulosic biomass into advanced functional materials and their potential applications. Lignocellulosic biomass represents an as-of-yet underutilized renewable source for not only biofuel production but also functional materials fabrication. This renewable source is a great alternative for fossil fuel based chemicals, which could be one of the solutions to energy crisis. In this work, it was demonstrated a variety of advanced materials including functional carbons, metal and silica nanoparticles could be derived from lignocellulosic biomass. Chapter 1 provided overall reviewed of the lignin structures, productions and its utilizations as plastics, absorbents and carbons, as well as the preparation of nano-structured silver, silica and silicon carbide/nitride from biomass. Chapter 2, 3 and 4 discussed the fabrication of highly porous carbons from isolated lignin, and their applications as electric supercapacitors for energy storage. In chapter 2, ultrafine porous carbon fibers were prepared via electrospinning followed by simultaneous carbonization and activation. Chapter 3 covered the fabrication of supercapacitor based on the porous carbon fibers and the investigation of their electrochemical performances. In chapter 4, porous carbon particulates with layered carbon nano plates structures were produced by simple oven-drying followed by simultaneous carbonization and activation. The effects of heat processing parameters on the resulting carbon structures and their electrochemical properties were discussed in details. Chapter 5 and 6 addressed the preparation of silver nanoparticles using lignin. Chapter 5 reported the synthesis, underlying kinetics and mechanism of monodispersed silver nanospheres with diameter less than 25 nm in aqueous solutions using lignin as dual reducing and capping agents. Chapter 6 covered the preparation of silver nanoparticles on electrospun celluloses ultrafine fibers using lignin as both binding and reducing agents. The efficiency of this synthetic protocol and the properties of resulting particles were examined. Chapter 7 reported the streamlined extraction of lignin/hemicelluloses and silica from rice straw and their subsequent conversion to activated carbon and monodispersed silica particles.

Types of Faculty Scholars in Community Colleges

ERIC Educational Resources Information Center

Park, Toby J.; Braxton, John M.; Lyken-Segosebe, Dawn

2015-01-01

This chapter describes three empirically derived types of faculty scholars in community colleges: Immersed Scholars, Scholars of Dissemination, and Scholars of Pedagogical Knowledge. This chapter discusses these types and offers a recommendation.

FDA Bacteriological Analytical Manual, Chapter 10, 2003: Listeria monocytogenes

EPA Pesticide Factsheets

FDA Bacteriological Analytical Manual, Chapter 10 describes procedures for analysis of food samples and may be adapted for assessment of solid, particulate, aerosol, liquid and water samples containing Listeria monocytogenes.

Multidisciplinary Design Optimization for Aeropropulsion Engines and Solid Modeling/Animation via the Integrated Forced Methods

NASA Technical Reports Server (NTRS)

2004-01-01

The grant closure report is organized in the following four chapters: Chapter describes the two research areas Design optimization and Solid mechanics. Ten journal publications are listed in the second chapter. Five highlights is the subject matter of chapter three. CHAPTER 1. The Design Optimization Test Bed CometBoards. CHAPTER 2. Solid Mechanics: Integrated Force Method of Analysis. CHAPTER 3. Five Highlights: Neural Network and Regression Methods Demonstrated in the Design Optimization of a Subsonic Aircraft. Neural Network and Regression Soft Model Extended for PX-300 Aircraft Engine. Engine with Regression and Neural Network Approximators Designed. Cascade Optimization Strategy with Neural network and Regression Approximations Demonstrated on a Preliminary Aircraft Engine Design. Neural Network and Regression Approximations Used in Aircraft Design.

Gate-controlled-diodes in silicon-on-sapphire: A computer simulation

NASA Technical Reports Server (NTRS)

Gassaway, J. D.

1974-01-01

The computer simulation of the electrical behavior of a Gate-Controlled Diode (GCD) fabricated in Silicon-On-Sapphire (SOS) was described. A procedure for determining lifetime profiles from capacitance and reverse current measurements on the GCD was established. Chapter 1 discusses the SOS structure and points out the need of lifetime profiles to assist in device design for GCD's and bipolar transistors. Chapter 2 presents the one-dimensional analytical formula for electrostatic analysis of the SOS-GCD which are useful for data interpretation and setting boundary conditions on a simplified two-dimensional analysis. Chapter 3 gives the results of a two-dimensional analysis which treats the field as one-dimensional until the silicon film is depleted and the field penetrates the sapphire substrate. Chapter 4 describes a more complete two-dimensional model and gives results of programs implementing the model.

Hanford Site National Environmental Policy Act (NEPA) Characterization Report

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

Neitzel, Duane A.; Bunn, Amoret L.; Cannon, Sandra D.

2004-09-22

This document describes the U.S. Department of Energy's (DOE) Hanford Site environment. It is updated each year and is intended to provide a consistent description of the Hanford Site environment for the many National Environmental Policy Act (NEPA) documents being prepared by DOE contractors. No statements of significance or environmental consequences are provided. This year's report is the sixteenth revision of the original document published in 1988 and is (until replaced by the seventeenth revision) the only version that is relevant for use in the preparation of Hanford NEPA, State Environmental Policy Act (SEPA), and Comprehensive Environmental Response, Compensation, andmore » Liability Act (CERCLA) documents. The two chapters included in this document (Chapters 4 and 6) are numbered to correspond to the chapters where such information is typically presented in environmental impact statements (Weiss) and other Hanford Site NEPA or CERCLA documentation. Chapter 4.0 (Affected Environment) describes Hanford Site climate and meteorology, geology, hydrology, ecology, cultural, archaeological, and historical resources, socioeconomics, occupational safety and health, and noise. Chapter 6.0 (Statutory and Regulatory Requirements) describes federal and state laws and regulations, DOE directives and permits, and presidential executive orders that are applicable to the NEPA documents prepared for Hanford Site activities.« less

A Process Evaluation of Project Developmental Continuity. Interim Report II, Part B: Recommendations for Measuring Program Impact.

ERIC Educational Resources Information Center

Love, John M.; And Others

This report presents recommendations for measures to be used in assessing the impact of Project Developmental Continuity (PDC). Chapter I reviews the purpose of the impact study and presents the basic considerations guiding the selection of measures. Chapter II describes the review process that led to the final recommendations. Chapter III…

Do Pretests Increase Student Achievement as Measured by Posttests?

ERIC Educational Resources Information Center

Bancroft, Roger J.

This report describes a study of the effects of using pretests in science classes on chapter test achievement results. The targeted population consisted of eighth grade science students at a junior high school from 1992 to 2001. Whether giving a pretest followed by a posttest at the end of the chapter, or giving only the test at chapter end…

Developing an On-Line Textbook: Question-led Teaching and the World Wide Web.

ERIC Educational Resources Information Center

McCain, Roger A.

1999-01-01

Describes a demonstration project of a textbook chapter created with a vision for an active and interactive question-led textbook commenting on the content of the demonstration chapter, using HTML and Javascript in creating it, and aspects of the chapter as it is actually experienced. Includes reactions by Michael K. Salemi and Kailash Khandke.…

Research and Development Funding in the Proposed Fiscal Year 1985 Budget. Special Study.

ERIC Educational Resources Information Center

Schorsch, Louis; Dailey, Theresa

This report discusses in detail the research and development (R&D) elements of the Reagan Administration's budget for Fiscal Year (FY) 1985. Following an introduction (chapter I), chapter II presents an overview of R&D funding in the President's FY 1985 budget. The chapter begins by describing overall R&D spending in terms of the major catagories…

Educational Issues in Utah: Governance, Legislation, Technology, and Finance. 1994-95 Conditions of Education in Utah Yearbook.

ERIC Educational Resources Information Center

Galvin, Patrick F., Ed.; Johnson, Bob L., Jr., Ed.

This document is the third edition of "Conditions of Education in Utah," covering the 1994-95 academic year. The first three chapters analyze issues relative to distance education and the Internet. Chapters 1 and 2 examine the pros and cons of distance education, and chapter 3 describes the construction, maintenance, and staffing costs…

Chapter 10. Developing a habitat monitoring program: three examples from national forest planning

Treesearch

Michael I. Goldstein; Lowell H. Suring; Christina D. Vojta; Mary M. Rowland; Clinton. McCarthy

2013-01-01

This chapter reviews the process steps of wildlife habitat monitoring described in chapters 2 through 9 and provides three case examples that illustrate how the process steps apply to specific situations. It provides the reader an opportunity to synthesize the material while also revealing the potential knowledge gaps and pitfalls that may complicate completion of a...

Family Needs and Family Quality of Life for Taiwanese Families of Children with Intellectual Disability and Developmental Delay

ERIC Educational Resources Information Center

Chiu, Chun-Yu

2013-01-01

This dissertation consists of four related chapters including an introductory overview of all four chapters, a report on family needs, a report on family quality of life, and a summary of implications for the conceptual framework. Chapter 1, the introductory overview, presents background information of Taiwan and describes the family quality of…

Equality of Opportunity and Vocational Training Five Years on...Vocational Training Measures for Women in the European Community. Synthesis Report and Recommendations.

ERIC Educational Resources Information Center

Oels, Monika; Seeland, Suzanne

This document includes four chapters, a bibliography, notes, and two appendices. Chapter 1 briefly indicates the purpose of the report, to look retrospectively at innovative vocational training programs initiated for women in the European Community (EC). Chapter 2 describes general developments since 1978, including the legal background, the…

Toward Ubiquitous BCIs

NASA Astrophysics Data System (ADS)

Allison, Brendan Z.

The preceding chapters in this book described modern BCI systems. This concluding chapter instead discusses future directions. While there are some specific predictions, I mainly analyze key factors and trends relating to practical mainstream BCI development. While I note some disruptive technologies that could dramatically change BCIs, this chapter focuses mainly on realistic, incremental progress and how progress could affect user groups and ethical issues.

State School Finance Reform in the 1970's (Excerpted from School Finance Reform: A Legislators' Handbook).

ERIC Educational Resources Information Center

National Conference of State Legislatures, Washington, DC.

This report describes and evaluates the record of reform in several states that have revised their school finance systems since 1971. The introductory chapter provides an overview of reform's recent successes, shortcomings, and prospects. Six subsequent chapters offer a close look at its main features. Chapters 2 and 3 analyze new school aid…

Investigations of Galaxy Clusters Using Gravitational Lensing

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

Wiesner, Matthew P.

2014-08-01

In this dissertation, we discuss the properties of galaxy clusters that have been determined using strong and weak gravitational lensing. A galaxy cluster is a collection of galaxies that are bound together by the force of gravity, while gravitational lensing is the bending of light by gravity. Strong lensing is the formation of arcs or rings of light surrounding clusters and weak lensing is a change in the apparent shapes of many galaxies. In this work we examine the properties of several samples of galaxy clusters using gravitational lensing. In Chapter 1 we introduce astrophysical theory of galaxy clusters andmore » gravitational lensing. In Chapter 2 we examine evidence from our data that galaxy clusters are more concentrated than cosmology would predict. In Chapter 3 we investigate whether our assumptions about the number of galaxies in our clusters was valid by examining new data. In Chapter 4 we describe a determination of a relationship between mass and number of galaxies in a cluster at higher redshift than has been found before. In Chapter 5 we describe a model of the mass distribution in one of the ten lensing systems discovered by our group at Fermilab. Finally in Chapter 6 we summarize our conclusions.« less

SeaWiFS technical report series. Volume 28: SeaWiFS algorithms, part 1

NASA Technical Reports Server (NTRS)

Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor); Acker, James G. (Editor); Mcclain, Charles R.; Arrigo, Kevin; Esaias, Wayne E.; Darzi, Michael; Patt, Frederick S.; Evans, Robert H.; Brown, James W.

1995-01-01

This document provides five brief reports that address several algorithm investigations sponsored by the Calibration and Validation Team (CVT) within the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Project. This volume, therefore, has been designated as the first in a series of algorithm volumes. Chapter 1 describes the initial suite of masks, used to prevent further processing of contaminated radiometric data, and flags, which are employed to mark data whose quality (due to a variety of factors) may be suspect. In addition to providing the mask and flag algorithms, this chapter also describes the initial strategy for their implementation. Chapter 2 evaluates various strategies for the detection of clouds and ice in high latitude (polar and sub-polar regions) using Coastal Zone Color Scanner (CZCS) data. Chapter 3 presents an algorithm designed for detecting and masking coccolithosphore blooms in the open ocean. Chapter 4 outlines a proposed scheme for correcting the out-of-band response when SeaWiFS is in orbit. Chapter 5 gives a detailed description of the algorithm designed to apply sensor calibration data during the processing of level-1b data.

Computational Elucidation of Selectivities and Mechanisms Performed by Organometallic and Bioinorganic Catalysts

NASA Astrophysics Data System (ADS)

Grandner, Jessica Marie

Computational methods were used to determine the mechanisms and selectivities of organometallic-catalyzed reactions. The first half of the dissertation focuses on the study of metathesis catalysts in collaboration with the Grubbs group at CalTech. Chapter 1 describes the studies of the decomposition modes of several ruthenium-based metathesis catalysts. These studies were performed to better understand the decomposition of such catalysts in order to prevent decomposition (Chapter 1.2) or utilize decomposed catalysts for alternative reactions (Chapter 1.1). Chapter 2.1 describes the computational investigation of the origins of stereoretentive metathesis with ruthenium-based metathesis catalysts. These findings were then used to computationally design E-selective metathesis catalysts (Chapter 2.2). While the first half of the dissertation was centered around ruthenium catalysts, the second half of the dissertation pertains to iron-catalyzed reaction, in particular, iron-catalyzed reactions by P450 enzymes. The elements of Chapter 3 concentrate on the stereo- and chemo-selectivity of P450-catalyzed C-H hydroxylations. By combining multiple computational methods, the inherent activity of the iron-oxo catalyst and the influence of the active site on such reactions were illuminated. These discoveries allow for the engineering of new substrates and mutant enzymes for tailored C-H hydroxylation. While the mechanism of C-H hydroxylations catalyzed by P450 enzymes has been well studied, there are several P450-catalyzed transformations for which the mechanism is unknown. The components of Chapter 4 describe the use of computations to determine the mechanisms of complex, multi-step reactions catalyzed by P450s. The determination of these mechanisms elucidates how these enzymes react with various functional groups and substrate architectures and allows for a better understanding of how drug-like compounds may be broken down by human P450s.

Chapter 16: Retrocommissioning Evaluation Protocol. The Uniform Methods Project: Methods for Determining Energy Efficiency Savings for Specific Measures

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

Kurnik, Charles W.; Tiessen, Alex

Retrocommissioning (RCx) is a systematic process for optimizing energy performance in existing buildings. It specifically focuses on improving the control of energy-using equipment (e.g., heating, ventilation, and air conditioning [HVAC] equipment and lighting) and typically does not involve equipment replacement. Field results have shown proper RCx can achieve energy savings ranging from 5 percent to 20 percent, with a typical payback of two years or less (Thorne 2003). The method presented in this protocol provides direction regarding: (1) how to account for each measure's specific characteristics and (2) how to choose the most appropriate savings verification approach.

Mouse Models Applied to the Research of Pharmacological Treatments in Asthma.

PubMed

Marqués-García, Fernando; Marcos-Vadillo, Elena

2016-01-01

Models developed for the study of asthma mechanisms can be used to investigate new compounds with pharmacological activity against this disease. The increasing number of compounds requires a preclinical evaluation before starting the application in humans. Preclinical evaluation in animal models reduces the number of clinical trials positively impacting in the cost and in safety. In this chapter, three protocols for the study of drugs are shown: a model to investigate corticoids as a classical treatment of asthma; a protocol to test the effects of retinoic acid (RA) on asthma; and a mouse model to test new therapies in asthma as monoclonal antibodies.

High-throughput and reliable protocols for animal microRNA library cloning.

PubMed

Xiao, Caide

2011-01-01

MicroRNAs are short single-stranded RNA molecules (18-25 nucleotides). Because of their ability to silence gene expressions, they can be used to diagnose and treat tumors. Experimental construction of microRNA libraries was the most important step to identify microRNAs from animal tissues. Although there are many commercial kits with special protocols to construct microRNA libraries, this chapter provides the most reliable, high-throughput, and affordable protocols for microRNA library construction. The high-throughput capability of our protocols came from a double concentration (3 and 15%, thickness 1.5 mm) polyacrylamide gel electrophoresis (PAGE), which could directly extract microRNA-size RNAs from up to 400 μg total RNA (enough for two microRNA libraries). The reliability of our protocols was assured by a third PAGE, which selected PCR products of microRNA-size RNAs ligated with 5' and 3' linkers by a miRCat™ kit. Also, a MathCAD program was provided to automatically search short RNAs inserted between 5' and 3' linkers from thousands of sequencing text files.

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Volume 1; Revised

NASA Technical Reports Server (NTRS)

Mueller, James L. (Editor); Fargion, Giulietta (Editor); Mueller, J. L.; Trees, C.; Austin, R. W.; Pietras, C.; Hooker, S.; Holben, B.; McClain, Charles R.; Clark, D. K.;

Ocean Optics Protocols for Satellite Ocean Color Sensor Validation. Volume 2; Revised

NASA Technical Reports Server (NTRS)

Mueller, James L. (Editor); Fargion, Giulietta S. (Editor); Trees, C.; Austin, R. W.; Pietras, C. (Editor); Hooker, S.; Holben, B.; McClain, Charles R.; Clark, D. K.; Yuen, M.

2002-01-01

This document stipulates protocols for measuring bio-optical and radiometric data for the SIMBIOS Project. It supersedes the earlier version, and is organized into four parts: Introductory Background, Instrument Characteristics, Field Measurements and Data Analysis, Data Reporting and Archival. Changes in this revision include the addition of three new chapters: (1) Fundamental Definitions, Relationships and Conventions; (2) MOBY, A Radiometric Buoy for Performance Monitoring and Vicarious Calibration of Satellite Ocean Color Sensors: Measurement and Data Analysis Protocols; and (3) Normalized Water-Leaving Radiance and Remote Sensing Reflectance: Bidirectional Reflectance and Other Factors. Although the present document represents another significant, incremental improvement in the ocean optics protocols, there are several protocols that have either been overtaken by recent technological progress, or have been otherwise identified as inadequate. Revision 4 is scheduled for completion sometime in 2003. This technical report is not meant as a substitute for scientific literature. Instead, it will provide a ready and responsive vehicle for the multitude of technical reports issued by an operational Project. The contributions are published as submitted, after only minor editing to correct obvious grammatical or clerical errors.

Chapter 1: Reliably Measuring the Performance of Emerging Photovoltaic Solar Cells

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

Rumbles, Garry; Reese, Matthew O; Marshall, Ashley

Determining the power conversion efficiency of photovoltaic solar cells, especially those from new, emerging areas of technology, is important if advances in performance are to be made. However, although precise measurements are important, it is the accuracy of these types of measurements that can cause issues. Accurate measurements not only promote the development of new technology platforms, but they also enable comparisons with established technologies and allow assessments of advancements within the same field. This chapter provides insights into how measurements can be made with reasonable accuracy using both the components of the measuring system and a good protocol tomore » acquire good data. The chapter discusses how to measure a calibrated lamp spectrum, determine a spectral mismatch factor, identify the correct reference cell and filter, define the illuminated active area, measure J-V curves to avoid any hysteresis effects, take note of sample degradation issues and avoid the temptation to artificially enhance efficiency data.« less

Dynamics of an island agroecosystem: Where to now?

Treesearch

Marjorie V. C. Falanruw;  Francis Ruegorong

2015-01-01

This chapter describes past interaction of human populations with habitats on mainland Yap and the dynamics of the agroecosystem that developed. Recent changes in this system on mainland Yap are described, followed by a description of the nature and threats to agriculture in the Outer Islands. The chapter concludes by asking “Where to now?” and considers a past...

Wood-based composite materials : panel products, glued-laminated timber, structural composite lumber, and wood-nonwood composite materials

Treesearch

Nicole M. Stark; Zhiyong Cai; Charles Carll

2010-01-01

This chapter gives an overview of the general types and composition of wood-based composite products and the materials and processes used to manufacture them. It describes conventional wood-based composite panels and structural composite materials intended for general construction, interior use, or both. This chapter also describes wood–nonwood composites. Mechanical...

Child Support Enforcement (9th Annual Report to Congress for the Period Ending September 30, 1984).

ERIC Educational Resources Information Center

Office of Child Support Enforcement (DHHS), Washington, DC.

Described in this report are fiscal year 1984 activities of the Child Support Enforcement (CSE) program, a program established in 1975 as part D of title IV of the Social Security Act. Following an executive summary, chapter I describes the mission and organization of the CSE. Chapter II reviews the child support enforcement amendments of 1984,…

Higher Education and Employment in Japan: Trends and Issues. RIHE International Publication Series No. 5.

ERIC Educational Resources Information Center

Kaneko, Motohisa

This book describes and examines the crisis within higher education in Japan as it applies to what is taught as compared to the labor market needs of the country. The book is divided into three chapters. Chapter 1 describes the higher education system in Japan and its enrollment, the distribution of enrollment by field of study, and the flow of…

Finding the Higgs boson of the standard model in the channel ZH → e +e -b$$\\bar{b}$$ with the D0 detector at the Tevatron; Recherche du boson de Higgs du nideke standard dans le canal ZH → e +e -b$$\\bar{b}$$ avec le detecteur DØ aupres du Tevatron (in French)

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

Calpas, Betty Constante

2010-06-11

The organization of this thesis consists of three main ideas: the first presents the theoretical framework and experimental, as well as objects used in the analysis and the second relates to the various work tasks of service that I performed on the calorimeter, and the third is the search for the Higgs boson in the channel ZH → e +e -bmore » $$\\bar{b}$$. Thus, this thesis has the following structure: Chapter 1 is an introduction to the standard model of particle physics and the Higgs mechanism; Chapter 2 is an overview of the complex and the acceleration of the Tevatron at Fermilab DØ detector; Chapter 3 is an introduction to physical objects used in this thesis; Chapter 4 presents the study made on correcting the energy measured in the calorimeter; Chapter 5 describes the study of certification of electrons in the calorimeter; Chapter 6 describes the study of certification of electrons in the intercryostat region of calorimeter; Chapter 7 Detailed analysis on the search for Higgs production in the channel ZH → e +e -b$$\\bar{b}$$; and Chapter 8 presents the final results of the calculations of upper limits to the production cross section of the Higgs boson on a range of low masses.« less

Engineering of ribozyme-based aminoglycoside switches of gene expression by in vivo genetic selection in Saccharomyces cerevisiae.

PubMed

Klauser, Benedikt; Rehm, Charlotte; Summerer, Daniel; Hartig, Jörg S

2015-01-01

Synthetic RNA-based switches are a growing class of genetic controllers applied in synthetic biology to engineer cellular functions. In this chapter, we detail a protocol for the selection of posttranscriptional controllers of gene expression in yeast using the Schistosoma mansoni hammerhead ribozyme as a central catalytic unit. Incorporation of a small molecule-sensing aptamer domain into the ribozyme renders its activity ligand-dependent. Aptazymes display numerous advantages over conventional protein-based transcriptional controllers, namely, the use of little genomic space for encryption, their modular architecture allowing for easy reprogramming to new inputs, the physical linkage to the message to be controlled, and the ability to function without protein cofactors. Herein, we describe the method to select ribozyme-based switches of gene expression in Saccharomyces cerevisiae that we successfully implemented to engineer neomycin- and theophylline-responsive switches. We also highlight how to adapt the protocol to screen for switches responsive to other ligands. Reprogramming of the sensor unit and incorporation into any RNA of interest enables the fulfillment of a variety of regulatory functions. However, proper functioning of the aptazyme is largely dependent on optimal connection between the aptamer and the catalytic core. We obtained functional switches from a pool of variants carrying randomized connection sequences by an in vivo selection in MaV203 yeast cells that allows screening of a large sequence space of up to 1×10(9) variants. The protocol given explains how to construct aptazyme libraries, carry out the in vivo selection and characterize novel ON- and OFF-switches. © 2015 Elsevier Inc. All rights reserved.

PLGA microspheres encapsulating siRNA.

PubMed

De Rosa, Giuseppe; Salzano, Giuseppina

2015-01-01

The therapeutic use of small interfering RNA (siRNA) represents a new and powerful approach to suppress the expression of pathologically genes. However, biopharmaceutical drawbacks, such as short half-life, poor cellular uptake, and unspecific distribution into the body, hamper the development of siRNA-based therapeutics. Poly(lactide-co-glycolide), (PLGA) microspheres can be a useful tool to overcome these issues. siRNA can be encapsulated into the PLGA microspheres, which protects the loaded nucleic acid against the enzymatic degradation. Moreover, PLGA microspheres can be injected directly into the action site, where the siRNA can be released in controlled manner, thus avoiding the need of frequent invasive administrations. The complete biodegradability of PLGA to monomers easily metabolized by the body, and its approval by FDA and EMA for parenteral administration, assure the safety of this copolymer and do not require the removal of the device after the complete drug release. In chapter, a basic protocol for the preparation of PLGA microspheres encapsulating siRNA is described. This protocol is based on a double emulsion/solvent evaporation technique, a well known and easy to reproduce method. This specific protocol has been developed to encapsulate a siRNA anti-TNFα in PLGA microspheres, and it has been designed and optimized to achieve high siRNA encapsulation efficiency and slow siRNA release in vitro. However, it can be extended also to other siRNA as well as other RNA or DNA-based oligonucleotides (miRNA, antisense, decoy, etc.). Depending on the applications, chemical modifications of the backbone and site-specific modification within the siRNA sequences could be required.

Conceptual Chemical Process Design for Sustainability.

EPA Science Inventory

This chapter examines the sustainable design of chemical processes, with a focus on conceptual design, hierarchical and short-cut methods, and analyses of process sustainability for alternatives. The chapter describes a methodology for incorporating process sustainability analyse...

Pesticide Registration Manual: Chapter 12 - Applying for an Experimental Use Permit

EPA Pesticide Factsheets

This chapter of the pesticide registration manual describes the all requirements for Experimental Use Permits (EUP), and includes how to apply for an EUP, labeling requirements, application format, and data reporting requirements.

The vegetative communities associated with mammals of the South. Chapter 2

Treesearch

Beverly Collins; Philip E. Hyatt; Margaret K. Trani

2007-01-01

This chapter describes the ecoregions and vegetation types associated with mammals of the South. The distribution of mammals in the South reflects historic biogeographical processes as well as physiography and vegetation.

Hanford Waste Vitrification Plant technical manual

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

Larson, D.E.; Watrous, R.A.; Kruger, O.L.

1996-03-01

A key element of the Hanford waste management strategy is the construction of a new facility, the Hanford Waste Vitrification Plant (HWVP), to vitrify existing and future liquid high-level waste produced by defense activities at the Hanford Site. The HWVP mission is to vitrify pretreated waste in borosilicate glass, cast the glass into stainless steel canisters, and store the canisters at the Hanford Site until they are shipped to a federal geological repository. The HWVP Technical Manual (Manual) documents the technical bases of the current HWVP process and provides a physical description of the related equipment and the plant. Themore » immediate purpose of the document is to provide the technical bases for preparation of project baseline documents that will be used to direct the Title 1 and Title 2 design by the A/E, Fluor. The content of the Manual is organized in the following manner. Chapter 1.0 contains the background and context within which the HWVP was designed. Chapter 2.0 describes the site, plant, equipment and supporting services and provides the context for application of the process information in the Manual. Chapter 3.0 provides plant feed and product requirements, which are primary process bases for plant operation. Chapter 4.0 summarizes the technology for each plant process. Chapter 5.0 describes the engineering principles for designing major types of HWVP equipment. Chapter 6.0 describes the general safety aspects of the plant and process to assist in safe and prudent facility operation. Chapter 7.0 includes a description of the waste form qualification program and data. Chapter 8.0 indicates the current status of quality assurance requirements for the Manual. The Appendices provide data that are too extensive to be placed in the main text, such as extensive tables and sets of figures. The Manual is a revision of the 1987 version.« less

The NOvA Technical Design Report

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

Ayres, D.S.; Drake, G.R.; Goodman, M.C.

Technical Design Report (TDR) describes the preliminary design of the NOvA accelerator upgrades, NOvA detectors, detector halls and detector sites. Compared to the March 2006 and November 2006 NOvA Conceptual Design Reports (CDR), critical value engineering studies have been completed and the alternatives still active in the CDR have been narrowed to achieve a preliminary technical design ready for a Critical Decision 2 review. Many aspects of NOvA described this TDR are complete to a level far beyond a preliminary design. In particular, the access road to the NOvA Far Detector site in Minnesota has an advanced technical design atmore » a level appropriate for a Critical Decision 3a review. Several components of the accelerator upgrade and new neutrino detectors also have advanced technical designs appropriate for a Critical Decision 3a review. Chapter 1 is an Executive Summary with a short description of the NOvA project. Chapter 2 describes how the Fermilab NuMI beam will provide a narrow band beam of neutrinos for NOvA. Chapter 3 gives an updated overview of the scientific basis for the NOvA experiment, focusing on the primary goal to extend the search for {nu}{sub {mu}} {yields} {nu}{sub e} oscillations and measure the sin{sup 2}(2{theta}{sub 13}) parameter. This parameter has not been measured in any previous experiment and NOvA would extend the search by about an order of magnitude beyond the current limit. A secondary goal is to measure the dominant mode oscillation parameters, sin{sup 2}(2{theta}{sub 23}) and {Delta}m{sub 32}{sup 2} to a more precise level than previous experiments. Additional physics goals for NOvA are also discussed. Chapter 4 describes the Scientific Design Criteria which the Fermilab accelerator complex, NOvA detectors and NOvA detector sites must satisfy to meet the physics goals discussed in Chapter 3. Chapter 5 is an overview of the NOvA project. The changes in the design relative to the NOvA CDR are discussed. Chapter 6 summarizes the NOvA design performance relative to the Design Criteria set out in Chapter 4. Chapter 7 presents the Work Breakdown Structure dictionary at Level 3 and the Milestone dictionary. Chapters 8 through 17 then take each Level 2 WBS element of the NOvA project and present each part of the design in more detail than the overview given in Chapter 5. Specific technical design criteria are delineated for each part of the project in addition to the scientific design criteria outlined in Chapter 4. Changes in the design since the NOvA CDR are discussed in detail. The work remaining to bring each part of this preliminary design to a final design is outlined. Appendix A is a guide to other NOvA Project documentation with links to those documents.« less

Proper Plugin Protocols

DTIC Science & Technology

2011-12-28

specify collaboration constraints that occur in Java and XML frameworks and that the collaboration constraints from these frameworks matter in practice. (a...programming language boundaries, and Chapter 6 and Appendix A demonstrate that Fusion can specify constraints across both Java and XML in practice. (c...designed JUnit, Josh Bloch designed Java Collec- tions, and Krzysztof Cwalina designed the .NET Framework APIs. While all of these frameworks are very

Quasi-one dimensional (Q1D) nanostructures: Synthesis, integration and device application

NASA Astrophysics Data System (ADS)

Chien, Chung-Jen

Quasi-one-dimensional (Q1D) nanostructures such as nanotubes and nanowires have been widely regarded as the potential building blocks for nanoscale electronic, optoelectronic and sensing devices. In this work, the content can be divided into three categories: Nano-material synthesis and characterizations, alignment and integration, physical properties and application. The dissertation consists of seven chapters as following. Chapter 1 will give an introduction to low dimensional nano-materials. Chapter 2 explains the mechanism how Q1D nanostructure grows. Chapter 3 describes the methods how we horizontally and vertically align the Q1D nanostructure. Chapter 4 and 5 are the electrical and optical device characterization respectively. Chapter 6 demonstrates the integration of Q1D nanostructures and the device application. The last chapter will discuss the future work and conclusion of the thesis.

The northern pike, a prized native but disastrous invasive: Chapter 14

USGS Publications Warehouse

Rutz, David; Massengill, Robert L.; Sepulveda, Adam; Dunker, Kristine J.

2018-01-01

As the chapters in this book describe, the northern pike Esox lucius Linneaus, 1758 is a fascinating fish that plays an important ecological role in structuring aquatic communities (chapter 8), has the capacity to aid lake restoration efforts (chapter 11), and contributes substantially to local economies, both as a highlysought after sport fish (chapter 12) and as a commercial fishing resource (chapter 13). However, despite the magnificent attributes of this fish, there is another side to its story. Specifically, what happens when northern pike, a highly efficient predator, becomes established outside its natural range? To explore this question, this chapter will investigate observed consequences from many locations where northern pike (hereafter referred to as “pike”) have been introduced and discuss potential reasons why pike, under the right circumstances, can be considered an invasive species.

Born-Oppenheimer ab initio QM/MM Molecular Dynamics Simulations of Enzyme Reactions

PubMed Central

Zhou, Yanzi; Wang, Shenglong; Li, Yongle; Zhang, Yingkai

2016-01-01

There are two key requirements for reliably simulating enzyme reactions: one is a reasonably accurate potential energy surface to describe the bond forming/breaking process as well as to adequately model the heterogeneous enzyme environment; the other is to perform extensive sampling since an enzyme system consists of at least thousands of atoms and its energy landscape is very complex. One attractive approach to meet both daunting tasks is Born-Oppenheimer ab initio QM/MM molecular dynamics simulation (aiQM/MM-MD) with umbrella sampling. In this chapter, we describe our recently developed pseudobond Q-Chem–Amber interface, which employs a combined electrostatic-mechanical embedding scheme with periodic boundary condition and the particle mesh Ewald method for long-range electrostatics interactions. In our implementation, Q-Chem and the sander module of Amber are combined at the source code level without using system calls, and all necessary data communications between QM and MM calculations are achieved via computer memory. We demonstrate the applicability of this pseudobond Q-Chem–Amber interface by presenting two examples, one reaction in aqueous solution and one enzyme reaction. Finally, we describe our established aiQM/MM-MD enzyme simulation protocol, which has been successfully applied to study more than a dozen enzymes. PMID:27498636

Protocols for Molecular Dynamics Simulations of RNA Nanostructures.

PubMed

Kim, Taejin; Kasprzak, Wojciech K; Shapiro, Bruce A

2017-01-01

Molecular dynamics (MD) simulations have been used as one of the main research tools to study a wide range of biological systems and bridge the gap between X-ray crystallography or NMR structures and biological mechanism. In the field of RNA nanostructures, MD simulations have been used to fix steric clashes in computationally designed RNA nanostructures, characterize the dynamics, and investigate the interaction between RNA and other biomolecules such as delivery agents and membranes.In this chapter we present examples of computational protocols for molecular dynamics simulations in explicit and implicit solvent using the Amber Molecular Dynamics Package. We also show examples of post-simulation analysis steps and briefly mention selected tools beyond the Amber package. Limitations of the methods, tools, and protocols are also discussed. Most of the examples are illustrated for a small RNA duplex (helix), but the protocols are applicable to any nucleic acid structure, subject only to the computational speed and memory limitations of the hardware available to the user.

Analysis of Pervasive Mobile Ad Hoc Routing Protocols

NASA Astrophysics Data System (ADS)

Qadri, Nadia N.; Liotta, Antonio

Mobile ad hoc networks (MANETs) are a fundamental element of pervasive networks and therefore, of pervasive systems that truly support pervasive computing, where user can communicate anywhere, anytime and on-the-fly. In fact, future advances in pervasive computing rely on advancements in mobile communication, which includes both infrastructure-based wireless networks and non-infrastructure-based MANETs. MANETs introduce a new communication paradigm, which does not require a fixed infrastructure - they rely on wireless terminals for routing and transport services. Due to highly dynamic topology, absence of established infrastructure for centralized administration, bandwidth constrained wireless links, and limited resources in MANETs, it is challenging to design an efficient and reliable routing protocol. This chapter reviews the key studies carried out so far on the performance of mobile ad hoc routing protocols. We discuss performance issues and metrics required for the evaluation of ad hoc routing protocols. This leads to a survey of existing work, which captures the performance of ad hoc routing algorithms and their behaviour from different perspectives and highlights avenues for future research.

5 CFR Appendix B to 5 Cfr Chapter... - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2013 CFR

2013-01-01

... Assigned Responsibilities of the General Counsel of the Federal Labor Relations Authority B Appendix B to 5... FEDERAL LABOR RELATIONS AUTHORITY AND FEDERAL SERVICE IMPASSES PANEL Ch. XIV, App. B Appendix B to 5 CFR... chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other powers of the...

5 CFR Appendix B to 5 Cfr Chapter... - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2014 CFR

2014-01-01

... Assigned Responsibilities of the General Counsel of the Federal Labor Relations Authority B Appendix B to 5... FEDERAL LABOR RELATIONS AUTHORITY AND FEDERAL SERVICE IMPASSES PANEL Ch. XIV, App. B Appendix B to 5 CFR... chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other powers of the...

5 CFR Appendix B to 5 Cfr Chapter... - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2012 CFR

2012-01-01

... Assigned Responsibilities of the General Counsel of the Federal Labor Relations Authority B Appendix B to 5... FEDERAL LABOR RELATIONS AUTHORITY AND FEDERAL SERVICE IMPASSES PANEL Ch. XIV, App. B Appendix B to 5 CFR... chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other powers of the...

5 CFR Appendix B to 5 Cfr Chapter... - Memorandum Describing the Authority and Assigned Responsibilities of the General Counsel of the...

Code of Federal Regulations, 2011 CFR

2011-01-01

... Assigned Responsibilities of the General Counsel of the Federal Labor Relations Authority B Appendix B to 5... FEDERAL LABOR RELATIONS AUTHORITY AND FEDERAL SERVICE IMPASSES PANEL Ch. XIV, App. B Appendix B to 5 CFR... chapter, (B) file and prosecute complaints under this chapter, and (C) exercise such other powers of the...

The Education of Adults in Britain: A Study of Organisation, Finance and Policy. Newland Papers Number Ten.

ERIC Educational Resources Information Center

Jennings, Bernard

This report updates a 1981 paper on adult education in Britain that was part of a series published by the European Centre for Leisure and Education. It includes five chapters. Chapter 1 provides statistics on the population and labor force of Britain and describes its education and government systems. Chapter 2 provides a brief description of the…

Chinese Biogas Digester. A Potential Model for Small-Scale, Rural Applications. (A Manual for Construction and Operation). Reprint No. R-51.

ERIC Educational Resources Information Center

Nakagawa, Charles H.; Honquilada, Q. L.

This book provides the basic knowledge and guides for the construction and operation of a small-scale, family-size biogas unit. The first chapter discusses the benefits of biogas production and the Chinese biogas model. The second chapter shows the components, design formulas, and sizing units of the biogas model. Chapter 3 describes actual…

Pupil Exchange in the European Community Venice Colloquium (October 24-28, 1977). Collection Studies. Education Series No. 5.

ERIC Educational Resources Information Center

Commission des Communautes Europeennes (Luxembourg).

This publication contains the papers of the Venice Colloquium held in October, 1977 by educators from European countries to discuss the topic of visits and exchanges for pupils. There are seven chapters. Chapter one discusses the context and objectives of the colloquium. Chapter two describes pupil visits and exchanges in the European community.…

SeaWiFS Technical Report Series. Volume 41; Case Studies for SeaWiFS Calibration and Validation

NASA Technical Reports Server (NTRS)

Yeh, Eueng-nan; Barnes, Robert A.; Darzi, Michael; Kumar, Lakshmi; Early, Edward A.; Johnson, B. Carol; Mueller, James L.; Trees, Charles C.

1997-01-01

This document provides brief reports, or case studies, on a number of investigations sponsored by the Calibration and Validation Team (CVT) within the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Project. Chapter I describes the calibration and characterization of the GSFC sphere, which was used in the recent recalibration of the SeaWiFS instrument. Chapter 2 presents a revision of the diffuse attenuation coefficient, K(490), algorithm based on the SeaWiFS wavelengths. Chapter 3 provides an implementation scheme for an algorithm to remove out-of-band radiance when using a sensor calibration based on a finite width (truncated) spectral response function, e.g., between the 1% transmission points. Chapter 4 describes the implementation schemes for the stray light quality flag (local area coverage [LAC] and global area coverage [GAC]) and the LAC stray light correction.

Reynolds stress closure in jet flows using wave models

NASA Technical Reports Server (NTRS)

Morris, Philip J.

1990-01-01

A collection of papers is presented. The outline of this report is as follows. Chapter three contains a description of a weakly nonlinear turbulence model that was developed. An essential part of the application of such a closure scheme to general geometry jets is the solution of the local hydrodynamic stability equation for a given jet cross-section. Chapter four describes the conformal mapping schemes used to map such geometries onto a simple computational domain. Chapter five describes a solution of a stability problem for circular, elliptic, and rectangular geometries. In chapter six linear models for the shock shell structure in non-circular jets is given. The appendices contain reprints of papers also published during this study including the following topics: (1) instability of elliptic jets; (2) a technique for predicting the shock cell structure in non-circular jets using a vortex sheet model; and (3) the resonant interaction between twin supersonic jets.

Final Report for Dynamic Models for Causal Analysis of Panel Data. Methodological Overview. Part II, Chapter 1.

ERIC Educational Resources Information Center

Hannan, Michael T.

This technical document, part of a series of chapters described in SO 011 759, describes a basic model of panel analysis used in a study of the causes of institutional and structural change in nations. Panel analysis is defined as a record of state occupancy of a sample of units at two or more points in time; for example, voters disclose voting…

Plume Measurement System (PLUMES) Calibration Experiment

DTIC Science & Technology

1994-08-01

calibration chamber was lished and documented. To apply acoustic designed and built Particles were suspended technology to monitoring suspended sedi- in the...procedures are described in Chap- ter 2. Repeatability of the experiments and validity of the results are de - scribed in Chapter 3. In Chapter 4, the range...went into their design . The first two subsections give an overview of the calibration chamber and its characteristics. The remaining subsections describe

Introduction: The SERENITY vision

NASA Astrophysics Data System (ADS)

Maña, Antonio; Spanoudakis, George; Kokolakis, Spyros

In this chapter we present an overview of the SERENITY approach. We describe the SERENITY model of secure and dependable applications and show how it addresses the challenge of developing, integrating and dynamically maintaining security and dependability mechanisms in open, dynamic, distributed and heterogeneous computing systems and in particular Ambient Intelligence scenarios. The chapter describes the basic concepts used in the approach and introduces the different processes supported by SERENITY, along with the tools provided.

A Performance Comparison of Tree and Ring Topologies in Distributed System

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

Huang, Min

A distributed system is a collection of computers that are connected via a communication network. Distributed systems have become commonplace due to the wide availability of low-cost, high performance computers and network devices. However, the management infrastructure often does not scale well when distributed systems get very large. Some of the considerations in building a distributed system are the choice of the network topology and the method used to construct the distributed system so as to optimize the scalability and reliability of the system, lower the cost of linking nodes together and minimize the message delay in transmission, and simplifymore » system resource management. We have developed a new distributed management system that is able to handle the dynamic increase of system size, detect and recover the unexpected failure of system services, and manage system resources. The topologies used in the system are the tree-structured network and the ring-structured network. This thesis presents the research background, system components, design, implementation, experiment results and the conclusions of our work. The thesis is organized as follows: the research background is presented in chapter 1. Chapter 2 describes the system components, including the different node types and different connection types used in the system. In chapter 3, we describe the message types and message formats in the system. We discuss the system design and implementation in chapter 4. In chapter 5, we present the test environment and results, Finally, we conclude with a summary and describe our future work in chapter 6.« less

SeaWiFS technical report series. Volume 5: Ocean optics protocols for SeaWiFS validation

NASA Technical Reports Server (NTRS)

Mueller, James L.; Austin, Roswell W.; Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor)

1992-01-01

Protocols are presented for measuring optical properties, and other environmental variables, to validate the radiometric performance of the Sea-viewing Wide Field-of-view Sensor (SeaWiFS), and to develop and validate bio-optical algorithms for use with SeaWiFS data. The protocols are intended to establish foundations for a measurement strategy to verify the challenging SeaWiFS accuracy goals of 5 percent in water-leaving radiances and 35 percent in chlorophyll alpha concentration. The protocols first specify the variables which must be measured, and briefly review rationale. Subsequent chapters cover detailed protocols for instrument performance specifications, characterizing and calibration instruments, methods of making measurements in the field, and methods of data analysis. These protocols were developed at a workshop sponsored by the SeaWiFS Project Office (SPO) and held at the Naval Postgraduate School in Monterey, California (9-12 April, 1991). This report is the proceedings of that workshop, as interpreted and expanded by the authors and reviewed by workshop participants and other members of the bio-optical research community. The protocols are a first prescription to approach unprecedented measurement accuracies implied by the SeaWiFS goals, and research and development are needed to improve the state-of-the-art in specific areas. The protocols should be periodically revised to reflect technical advances during the SeaWiFS Project cycle.

The ABCs of an evolutionary education science: The academic, behavioral, and cultural implications of an evolutionary approach to education theory and practice

NASA Astrophysics Data System (ADS)

Kauffman, Rick, Jr.

Calls for improving research-informed policy in education are everywhere. Yet, while there is an increasing trend towards science-based practice, there remains little agreement over which of the sciences to consult and how to organize a collective effort between them. What Education lacks is a general theoretical framework through which policies can be constructed, implemented, and assessed. This dissertation submits that evolutionary theory can provide a suitable framework for coordinating educational policies and practice, and can provide the entire field of education with a clearer sense of how to better manage the learning environment. This dissertation explores two broad paths that outline the conceptual foundations for an Evolutionary Education Science: "Teaching Evolution" and "Using Evolution to Teach." Chapter 1 introduces both of these themes. After describing why evolutionary science is best suited for organizing education research and practice, Chapter 1 proceeds to "teach" an overview of the "evolutionary toolkit"---the mechanisms and principles that underlie the modern evolutionary perspective. The chapter then employs the "toolkit" in examining education from an evolutionary perspective, outlining the evolutionary precepts that can guide theorizing and research in education, describing how educators can "use evolution to teach.". Chapters 2-4 expand on this second theme. Chapters 2 and 3 describe an education program for at-risk 9th and 10th grade students, the Regents Academy, designed entirely with evolutionary principles in mind. The program was rigorously assessed in a randomized control design and has demonstrated success at improving students' academic performance (Chapter 2) and social & behavioral development (Chapter 3). Chapter 4 examines current teaching strategies that underlie effective curriculum-instruction-assessment practices and proposes a framework for organizing successful, evidence-based strategies for neural-/cognitive-focused learning goals. Chapter 5 explores the cognitive effects that "teaching evolution" has on the learner. This chapter examines the effects that a course on evolutionary theory has on university undergraduate students in understanding and applying evolution and how learning the evolutionary toolkit affects critical thinking skills and domain transfer of knowledge. The results demonstrate that a single course on evolutionary theory increases students' acceptance and understanding of evolution and science, and, in effect, increases critical thinking performance.

Strategic Integration of Open Educational Resources in Higher Education

NASA Astrophysics Data System (ADS)

Schaffert, Sandra

Open Educational Resources (OERs) can be seen as social movement but are also implemented as strategic measures in higher education institutions (HEIs). This chapter describes the current aims and experiences of OERs in HEIs. Starting with definitions and milestones in respect of the current status, this chapter gives an overview of projects and implementation objectives and it describes two concrete case studies, i.e., the Massachusetts Institute of Technology Open CourseWare project and the OpenLearn project at the Open University in the United Kingdom. The aim of this chapter is to give a comprehensive overview to decision makers and policy drivers within higher education organizations, and thus it develops a blueprint of an implementation model.

Realising the Potential of Serenity in Emerging AmI Ecosystems:Implications and Challenges

NASA Astrophysics Data System (ADS)

Armenteros, Álvaro; García, Laura; Muñoz, Antonio; Maña, Antonio

In this chapter we describe the potential of SERENITY in Ambient Intelligence (AmI) Ecosystems. As a proof of concept, we describe the implementation of a prototype based on the application of the SERENITY model (including processes, artefacts and tools) to an industrial AmI scenario. A complete description of this prototype, along with all Security and Dependability (S&D) artefacts used is provided in this chapter. Besides, the chapter also provides a complete description of one of the S&D Patterns used in the prototype, in order both, to give a global view of SERENITY and to provide useful details about the realization of this scenario.

Fourth - generation languages. Volume 2. Representative 4GLs

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

Martin, J.

1986-01-01

The chapters in Part II each describe one representative product marketed by a vendor other than IBM. (Volume III of this work covers IBM languages.) The chapters in Part II cover the following 4GLs: ADS/ONLINE, APPLICATION FACTORY, DATATRIEVE, FOCUS, IDEAL, INTELLECT, MANTIS, MIMER, NATURAL, NOMAD2, RAMIS II, SYSTEM W, and USE-IT. The Perspective section of Part II presents a general overview of the product and describes its role in the marketplace. The Tutorial section describes how a user employs the language in application development.

One Billion Illiterates. One Billion Reasons for Action. Report and Extracts from Papers of an International Seminar on Co-operating for Literacy (Berlin, West Germany, October 1983).

ERIC Educational Resources Information Center

Fordham, Paul, Ed.

This document contains the report and edited papers from an international seminar emphasizing the sharing of ideas and resources to eliminate illiteracy. Chapter I of the seven-chapter report offers background information and seminar objectives. Chapter II provides a world overview, which describes the general development context of literacy. In…

A Guide to IRUS-II Application Development

DTIC Science & Technology

1989-09-01

Stallard (editors). Research and Develo; nent in Natural Language b’nderstan,;ng as Part of t/i Strategic Computing Program . chapter 3, pages 27-34...Development in Natural Language Processing in the Strategic Computing Program . Compi-nrional Linguistics 12(2):132-136. April-June, 1986. [24] Sidner. C.L...assist developers interested in adapting IRUS-11 to new application domains Chapter 2 provides a general introduction and overviev ,. Chapter 3 describes

20 CFR 416.925 - Listing of Impairments in appendix 1 of subpart P of part 404 of this chapter.

Code of Federal Regulations, 2011 CFR

2011-04-01

... subpart P of part 404 of this chapter. 416.925 Section 416.925 Employees' Benefits SOCIAL SECURITY... Blindness Medical Considerations § 416.925 Listing of Impairments in appendix 1 of subpart P of part 404 of... listings) is in appendix 1 of subpart P of part 404 of this chapter. For adults, it describes for each of...

20 CFR 416.925 - Listing of Impairments in appendix 1 of subpart P of part 404 of this chapter.

Code of Federal Regulations, 2012 CFR

2012-04-01

... subpart P of part 404 of this chapter. 416.925 Section 416.925 Employees' Benefits SOCIAL SECURITY... Blindness Medical Considerations § 416.925 Listing of Impairments in appendix 1 of subpart P of part 404 of... listings) is in appendix 1 of subpart P of part 404 of this chapter. For adults, it describes for each of...

Direct observation and control of hydrogen-bond dynamics using low-temperature scanning tunneling microscopy

NASA Astrophysics Data System (ADS)

Kumagai, Takashi

2015-08-01

Hydrogen(H)-bond dynamics are involved in many elementary processes in chemistry and biology. Because of its fundamental importance, a variety of experimental and theoretical approaches have been employed to study the dynamics in gas, liquid, solid phases, and their interfaces. This review describes the recent progress of direct observation and control of H-bond dynamics in several model systems on a metal surface by using low-temperature scanning tunneling microscopy (STM). General aspects of H-bond dynamics and the experimental methods are briefly described in chapter 1 and 2. In the subsequent four chapters, I present direct observation of an H-bond exchange reaction within a single water dimer (chapter 3), a symmetric H bond (chapter 4) and H-atom relay reactions (chapter 5) within water-hydroxyl complexes, and an intramolecular H-atom transfer reaction (tautomerization) within a single porphycene molecule (chapter 6). These results provide novel microscopic insights into H-bond dynamics at the single-molecule level, and highlight significant impact on the process from quantum effects, namely tunneling and zero-point vibration, resulting from the small mass of H atom. Additionally, local environmental effect on H-bond dynamics is also examined by using atom/molecule manipulation with the STM.

Regional demand forecasting and simulation model: user's manual. Task 4, final report

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

Parhizgari, A M

1978-09-25

The Department of Energy's Regional Demand Forecasting Model (RDFOR) is an econometric and simulation system designed to estimate annual fuel-sector-region specific consumption of energy for the US. Its purposes are to (1) provide the demand side of the Project Independence Evaluation System (PIES), (2) enhance our empirical insights into the structure of US energy demand, and (3) assist policymakers in their decisions on and formulations of various energy policies and/or scenarios. This report provides a self-contained user's manual for interpreting, utilizing, and implementing RDFOR simulation software packages. Chapters I and II present the theoretical structure and the simulation of RDFOR,more » respectively. Chapter III describes several potential scenarios which are (or have been) utilized in the RDFOR simulations. Chapter IV presents an overview of the complete software package utilized in simulation. Chapter V provides the detailed explanation and documentation of this package. The last chapter describes step-by-step implementation of the simulation package using the two scenarios detailed in Chapter III. The RDFOR model contains 14 fuels: gasoline, electricity, natural gas, distillate and residual fuels, liquid gases, jet fuel, coal, oil, petroleum products, asphalt, petroleum coke, metallurgical coal, and total fuels, spread over residential, commercial, industrial, and transportation sectors.« less

Effects of Heat Treatments and Compositional Modification on Carbide Network and Matrix Microstructure in Ultrahigh Carbon Steels

NASA Astrophysics Data System (ADS)

Hecht, Matthew David

This dissertation investigates microstructure/property relations in ultrahigh carbon steel (UHCS) with the aim of improving toughness while retaining high hardness. Due to high C contents (ranging from 1 to 2 wt%), UHCS exhibit high strength, hardness, and wear resistance. Despite this, applications for UHCS are currently limited because they typically contain a continuous network of proeutectoid cementite that greatly reduces ductility and toughness. In previous research, thermomechanic processing had seen considerable success in breaking up the network. However, the processing is difficult and has thus far seen very limited industrial application. Chemical modification of the steel composition has also seen some success in network break-up, but is still not well understood. There have been relatively few fundamental studies of microstructure evolution in UHCS; studies in the literature typically focused on lower C steels (up to 1 wt% C) or on cast irons (>2.1 wt% C). Thus, this work was undertaken to gain a better understanding of microstructural changes that occur during heat treatment and/or chemical modification of UHCS with a focus on the distribution of proeutectoid cementite within the microstructure. This dissertation is composed of eight chapters. The first chapter presents an introduction to phases found in UHCS, descriptions of research materials used in each chapter, and the hypotheses and objectives guiding the work. The second chapter describes a study of the microstructure found in a 2C-4Cr UHCS before and after an industrial-scale austenitizating heat treatment that increased hardness and toughness and also produced discrete carbide particles in the matrix. The third chapter establishes and demonstrates a metric for measuring connectivity in carbide networks. The fourth chapter describes a series of heat treatments designed to investigate kinetics of spheroidization and coarsening of carbide particles and denuded zones near cementite network branches in 2C-4Cr UHCS. The fifth chapter describes an additional series of heat treatments comparing coarsening kinetics in 2C-1Cr and 2C-4Cr UHCS. Lowering the Cr content caused clustering of cementite particles near grain boundaries, in contrast to the denuded zones observed in the higher Cr UHCS. The fifth chapter details four in situ confocal laser scanning microscopy heat treatments of 2C-4Cr UHCS. The seventh chapter investigates the effects of a 2wt% Nb addition on 2C-4Cr UHCS. The eighth and final chapter summarizes the findings of all the experiments of the previous chapters and revisits the objectives and conclusions.

The actin cytoskeleton in whole mount preparations and sections.

PubMed

Resch, Guenter P; Urban, Edit; Jacob, Sonja

2010-01-01

In non-muscle cells, the actin cytoskeleton plays a key role by providing a scaffold contributing to the definition of cell shape, force for driving cell motility, cytokinesis, endocytosis, and propulsion of pathogens, as well as tracks for intracellular transport. A thorough understanding of these processes requires insight into the spatial and temporal organisation of actin filaments into diverse higher-order structures, such as networks, parallel bundles, and contractile arrays. Transmission and scanning electron microscopy can be used to visualise the actin cytoskeleton, but due to the delicate nature of actin filaments, they are easily affected by standard preparation protocols, yielding variable degrees of ultrastructural preservation. In this chapter, we describe different conventional and cryo-approaches to visualise the actin cytoskeleton using transmission electron microscopy and discuss their specific advantages and drawbacks. In the first part, we present three different whole mount techniques, which allow visualisation of actin in the peripheral, thinly spread parts of cells grown in monolayers. In the second part, we describe specific issues concerning the visualisation of actin in thin sections. Techniques for three-dimensional visualisation of actin, protein localisation, and correlative light and electron microscopy are also included. Copyright © 2010 Elsevier Inc. All rights reserved.

A Simple XML Producer-Consumer Protocol

NASA Technical Reports Server (NTRS)

Smith, Warren; Gunter, Dan; Quesnel, Darcy

2000-01-01

This document describes a simple XML-based protocol that can be used for producers of events to communicate with consumers of events. The protocol described here is not meant to be the most efficient protocol, the most logical protocol, or the best protocol in any way. This protocol was defined quickly and it's intent is to give us a reasonable protocol that we can implement relatively easily and then use to gain experience in distributed event services. This experience will help us evaluate proposals for event representations, XML-based encoding of information, and communication protocols. The next section of this document describes how we represent events in this protocol and then defines the two events that we choose to use for our initial experiments. These definitions are made by example so that they are informal and easy to understand. The following section then proceeds to define the producer-consumer protocol we have agreed upon for our initial experiments.

Software Assurance Curriculum Project Volume 1: Master of Software Assurance Reference Curriculum

DTIC Science & Technology

2010-08-01

activity by providing a check on the relevance and currency of the process used to develop the MSwA2010 curriculum content. Figure 2 is an expansion of...random oracle model, symmetric crypto primitives, modes of operations, asymmetric crypto primitives (Chapter 5) [16] Detailed design...encryption, public key encryption, digital signatures, message authentication codes, crypto protocols, cryptanalysis, and further detailed crypto

Dynamic UNITY

DTIC Science & Technology

2002-01-01

UNITY program that implements exactly the same algorithm as Specification 1.1. The correctness of this program is proven in amanner sim- 4 program...chapter, we introduce the Dynamic UNITY formalism, which allows us to reason about algorithms and protocols in which the sets of participating processes...implements Euclid’s algorithm for calculating the greatest common divisor (GCD) of two integers; it repeat- edly reads an integer message from each of its

Chapter A7. Section 7.2. Fecal Indicator Viruses

USGS Publications Warehouse

Bushon, Rebecca N.

2003-01-01

More than 100 types of human pathogenic viruses may be present in fecal-contaminated waters. Coliphages are used as indicators of virus-related fecal contamination and of the microbiological quality of waters. This report provides information on the equipment, sampling protocols, and laboratory methods that are in standard use by U.S. Geological Survey (USGS) personnel for the collection of data on fecal indicator viruses.

It is Time the United States Air Force Changes the way it Feeds its Airmen

DTIC Science & Technology

2008-03-01

narrative , phenomenology , ethnography , case study , and grounded theory . In purpose, these strategies are...methodology) the research will be analyzed. Methodology A qualitative research methodology and specifically a case study strategy for the...well as theory building in chapter five . Finally, in regards to reliability, Yin’s (2003) case study protocol guidance was used as a means to

The Realm of Physical Geography.

ERIC Educational Resources Information Center

Rea, Patrick S.

This secondary education student geography book contains chapters on climate, landforms, oceans, world vegetation, water resources, and population. Each chapter provides an introduction that describes the unit's topics, descriptive and instructional materials, learning activities, and questions. A glossary of geography-related terms and an…

Overview of Decoding across the Disciplines

ERIC Educational Resources Information Center

Boman, Jennifer; Currie, Genevieve; MacDonald, Ron; Miller-Young, Janice; Yeo, Michelle; Zettel, Stephanie

2017-01-01

In this chapter we describe the Decoding the Disciplines Faculty Learning Community at Mount Royal University and how Decoding has been used in new and multidisciplinary ways in the various teaching, curriculum, and research projects that are presented in detail in subsequent chapters.

Reproducible Bioinformatics Research for Biologists

USDA-ARS?s Scientific Manuscript database

This book chapter describes the current Big Data problem in Bioinformatics and the resulting issues with performing reproducible computational research. The core of the chapter provides guidelines and summaries of current tools/techniques that a noncomputational researcher would need to learn to pe...

Chapter 9. Benefits of International Collaboration

EPA Science Inventory

In this chapter, we share what we have learned from working with our Brazilian colleagues on a multi university, multiyear, and multi basin ecological assessment and how those experiences were transmitted more broadly. These lessons (each of which is described in subsequent parag...

MONITORING OF INCINERATOR EMISSIONS

EPA Science Inventory

Monitoring of Incinerator Emissions is a chapter to be included in a book entitled Hazardous Waste Incineration, edited by A. Sarofim and D. Pershing, and published by John Wiley and Sons. he chapter describes stack sampling and analysis procedures in use on hazardous waste incin...

Chapter 10:Hardwoods for timber bridges

Treesearch

James P. Wacker; Ed T. Cesa

2005-01-01

This chapter describes the joint efforts of the Forest Service and the FHWA to administer national programs including research, demonstration bridges, and technology transfer components. Summary information on a number of Forest Service-WIT demonstration bridges constructed with hardwoods is also provided.

Laboratory Testing Protocols for Heparin-Induced Thrombocytopenia (HIT) Testing.

PubMed

Lau, Kun Kan Edwin; Mohammed, Soma; Pasalic, Leonardo; Favaloro, Emmanuel J

2017-01-01

Heparin-induced thrombocytopenia (HIT) represents a significant high morbidity complication of heparin therapy. The clinicopathological diagnosis of HIT remains challenging for many reasons; thus, laboratory testing represents an important component of an accurate diagnosis. Although there are many assays available to assess HIT, these essentially fall into two categories-(a) immunological assays, and (b) functional assays. The current chapter presents protocols for several HIT assays, being those that are most commonly performed in laboratory practice and have the widest geographic distribution. These comprise a manual lateral flow-based system (STiC), a fully automated latex immunoturbidimetric assay, a fully automated chemiluminescent assay (CLIA), light transmission aggregation (LTA), and whole blood aggregation (Multiplate).

Preparation of Stable Amyloid-β Oligomers Without Perturbative Methods.

PubMed

Kotler, Samuel A; Ramamoorthy, Ayyalusamy

2018-01-01

Soluble amyloid-β (Aβ) oligomers have become a focal point in the study of Alzheimer's disease due to their ability to elicit cytotoxicity. A number of recent studies have concentrated on the structural characterization of soluble Aβ oligomers to gain insight into their mechanism of toxicity. Consequently, providing reproducible protocols for the preparation of such oligomers is of utmost importance. The method presented in this chapter details a protocol for preparing an Aβ oligomer, with a primarily disordered secondary structure, without the need for chemical modification or amino acid substitution. Due to the stability of these disordered Aβ oligomers and the reproducibility with which they form, they are amenable for biophysical and high-resolution structural characterization.

In Vitro Culturing and Live Imaging of Drosophila Egg Chambers: A History and Adaptable Method.

PubMed

Peters, Nathaniel C; Berg, Celeste A

2016-01-01

The development of the Drosophila egg chamber encompasses a myriad of diverse germline and somatic events, and as such, the egg chamber has become a widely used and influential developmental model. Advantages of this system include physical accessibility, genetic tractability, and amenability to microscopy and live culturing, the last of which is the focus of this chapter. To provide adequate context, we summarize the structure of the Drosophila ovary and egg chamber, the morphogenetic events of oogenesis, the history of egg-chamber live culturing, and many of the important discoveries that this culturing has afforded. Subsequently, we discuss various culturing methods that have facilitated analyses of different stages of egg-chamber development and different types of cells within the egg chamber, and we present an optimized protocol for live culturing Drosophila egg chambers.We designed this protocol for culturing late-stage Drosophila egg chambers and live imaging epithelial tube morphogenesis, but with appropriate modifications, it can be used to culture egg chambers of any stage. The protocol employs a liquid-permeable, weighted "blanket" to gently hold egg chambers against the coverslip in a glass-bottomed culture dish so the egg chambers can be imaged on an inverted microscope. This setup provides a more buffered, stable, culturing environment than previously published methods by using a larger volume of culture media, but the setup is also compatible with small volumes. This chapter should aid researchers in their efforts to culture and live-image Drosophila egg chambers, further augmenting the impressive power of this model system.

In vitro culturing and live imaging of Drosophila egg chambers: A history and adaptable method

PubMed Central

Peters, Nathaniel C.; Berg, Celeste A.

2017-01-01

Summary/Abstract The development of the Drosophila egg chamber encompasses a myriad of diverse germline and somatic events, and as such, the egg chamber has become a widely used and influential developmental model. Advantages of this system include physical accessibility, genetic tractability, and amenability to microscopy and live culturing, the last of which is the focus of this chapter. To provide adequate context, we summarize the structure of the Drosophila ovary and egg chamber, the morphogenetic events of oogenesis, the history of egg-chamber live culturing, and many of the important discoveries that this culturing has afforded. Subsequently, we discuss various culturing methods that have facilitated analyses of different stages of egg-chamber development and different types of cells within the egg chamber, and we present an optimized protocol for live culturing Drosophila egg chambers. We designed this protocol for culturing late-stage Drosophila egg chambers and live imaging epithelial tube morphogenesis, but with appropriate modifications it can be used to culture egg chambers of any stage. The protocol employs a liquid-permeable, weighted, “blanket” to gently hold egg chambers against the coverslip in a glass-bottomed culture dish so the egg chambers can be imaged on an inverted microscope. This setup provides a more buffered, stable culturing environment than previously published methods by using a larger volume of culture media, but the setup is also compatible with small volumes. This chapter should aid researchers in their efforts to culture and live image Drosophila egg chambers, further augmenting the impressive power of this model system. PMID:27557572

Employee Relations. A Guide and Reference Book for Those Involved or Training to Be Involved in Employee Relations in the Hotel and Catering Industry. Seventh Edition.

ERIC Educational Resources Information Center

Martin, Rodney; Hayter, Roy, Ed.

This guide and reference book is designed to help those involved or training to be involved in employee relations in the hotel and catering industry. Chapter 1 attempts to define employee relations. Chapter 2 describes the institutions and parties involved in employee relations in the hotel and catering industry. The focus of chapter 3 is on…

Motivating Behavioral Change. Revised.

ERIC Educational Resources Information Center

Bricker, William A.; Campbell, Philippa H.

The manual describes approaches the teacher or therapist can use to motivate handicapped students. Chapter 1 considers motivation in terms of the consequences of an action, contingency awareness, and types of motivating conditions. A second chapter offers guidelines for identifying potential reinforcers and discusses examples of motivating…

The science of cheese

USDA-ARS?s Scientific Manuscript database

The book describes the science of cheese in everyday language. The first chapters cover milk, mammals, and principles of cheesemaking and aging, along with lactose intolerance and raw milk cheese. Succeeding chapters deal with a category of cheese along with a class of compounds associated with it...

Solid Lubrication Fundamentals and Applications. Chapter 1; Introduction and Background

NASA Technical Reports Server (NTRS)

Miyoshi, Kazuhisa

1996-01-01

This chapter presents an introduction and historical background to the field of tribology, especially solid lubrication and lubricants and sets them in the perspective of techniques and materials in lubrication. Also, solid and liquid lubrication films are defined and described.

Evaluation Methods Sourcebook.

ERIC Educational Resources Information Center

Love, Arnold J., Ed.

The chapters commissioned for this book describe key aspects of evaluation methodology as they are practiced in a Canadian context, providing representative illustrations of recent developments in evaluation methodology as it is currently applied. The following chapters are included: (1) "Program Evaluation with Limited Fiscal and Human…

Sex-Role Psychology.

ERIC Educational Resources Information Center

Wesley, Frank; Wesley, Claire

The book explores the psychological aspects of sex-role development and investigates various approaches which have been suggested to provide greater equality between females and males. Chapter one describes the psychology of women and identifies possible approaches toward integration of male and female characteristics. Chapter two discusses…

Baculoviridae

USDA-ARS?s Scientific Manuscript database

This book chapter describes the nomenclature and organization of the insect virus family Baculoviridae, and is an update of the same chapter from the previously published 8th Report of the International Committee on the Taxonomy of Viruses. Since the publication of the 8th report, the genomes of ba...

In vitro propagation of olive (Olea europaea L.) by nodal segmentation of elongated shoots.

PubMed

Lambardi, Maurizio; Ozudogru, Elif Aylin; Roncasaglia, Romano

2013-01-01

Olive (Olea europaea L.), long-living, ever-green fruit tree of the Old World, has been part of a traditional landscape in the Mediterranean area for centuries. Both the fruits consumed after processing and the oil extracted from the fruits are among the main components of the Mediterranean diet, widely used for salads and cooking, as well as for preserving other food. Documentations show that the ancient use of this beautiful tree also includes lamp fuel production, wool treatment, soap production, medicine, and cosmetics. However, unlike the majority of the fruit species, olive propagation is still a laborious practice. As regards traditional propagation, rooting of cuttings and grafting stem segments onto rootstocks are possible, former being achieved only when the cuttings are collected in specific periods (spring or beginning of autumn), and latter only when skilled grafters are available. In both the cases, performance of the cultivars varies considerably. The regeneration of whole plants from ovules, on the other hand, is used only occasionally. Micropropagation of olive is not easy mainly due to explant oxidation, difficulties in explant disinfection, and labor-oriented establishment of in vitro shoot cultures. However today, the progress in micropropagation technology has made available the complete protocols for several Mediterranean cultivars. This chapter describes a micropropagation protocol based on the segmentation of nodal segments obtained from elongated shoots.

Surface modification and multiple exciton generation studies of lead(II) sulfide nanoparticles

NASA Astrophysics Data System (ADS)

Zemke, Jennifer M.

2011-12-01

Solar energy is a green alternative to fossil fuels but solar technologies to date have been plagued by low conversion efficiencies and high input costs making solar power inaccessible to much of the developing world. Semiconductor nanoparticles (NPs) may provide a route to efficient, economical solar devices through a phenomenon called multiple exciton generation (MEG). Through MEG, semiconductor NPs use a high-energy input photon to create more than one exciton (electron-hole pair) per photon absorbed, thereby exhibiting large photoconversion efficiencies. While MEG has been studied in many NP systems, and we understand some of the factors that affect MEG, a rigorous analysis of the NP-ligand interface with respect to MEG is missing. This dissertation describes how the NP ligand shell directly affects MEG and subsequent charge carrier recombination. Chapter I describes the motivation for studying MEG with respect to NP surface chemistry. Chapter II provides an in-depth overview of the transient absorption experiment used to measure MEG in the NP samples. Chapter III highlights the effect of oleic acid and sodium 2, 3-dimercaptopropane sulfonate on MEG in PbS NPs. The differences in carrier recombination were accounted for by two differences between these ligands: the coordinating atom and/or the secondary structure of the ligand. Because of these hypotheses, experiments were designed to elucidate the origin of these effects by controlling the NP ligand shell. Chapter IV details a viable synthetic route to thiol and amine-capped PbS NPs using sodium 3-mercaptopropane sulfonate as an intermediate ligand. With the versatile ligand exchange described in Chapter IV, the MEG yield and carrier recombination was investigated for ligands with varying headgroups but the same secondary structure. The correlation of ligand donor atom to MEG is outlined in Chapter V. Finally, Chapter VI discusses the conclusions and future outlook of the research reported in this dissertation. This dissertation includes previously published and unpublished co-authored material.

Modeling and Design of GaN High Electron Mobility Transistors and Hot Electron Transistors through Monte Carlo Particle-based Device Simulations

NASA Astrophysics Data System (ADS)

Soligo, Riccardo

In this work, the insight provided by our sophisticated Full Band Monte Carlo simulator is used to analyze the behavior of state-of-art devices like GaN High Electron Mobility Transistors and Hot Electron Transistors. Chapter 1 is dedicated to the description of the simulation tool used to obtain the results shown in this work. Moreover, a separate section is dedicated the set up of a procedure to validate to the tunneling algorithm recently implemented in the simulator. Chapter 2 introduces High Electron Mobility Transistors (HEMTs), state-of-art devices characterized by highly non linear transport phenomena that require the use of advanced simulation methods. The techniques for device modeling are described applied to a recent GaN-HEMT, and they are validated with experimental measurements. The main techniques characterization techniques are also described, including the original contribution provided by this work. Chapter 3 focuses on a popular technique to enhance HEMTs performance: the down-scaling of the device dimensions. In particular, this chapter is dedicated to lateral scaling and the calculation of a limiting cutoff frequency for a device of vanishing length. Finally, Chapter 4 and Chapter 5 describe the modeling of Hot Electron Transistors (HETs). The simulation approach is validated by matching the current characteristics with the experimental one before variations of the layouts are proposed to increase the current gain to values suitable for amplification. The frequency response of these layouts is calculated, and modeled by a small signal circuit. For this purpose, a method to directly calculate the capacitance is developed which provides a graphical picture of the capacitative phenomena that limit the frequency response in devices. In Chapter 5 the properties of the hot electrons are investigated for different injection energies, which are obtained by changing the layout of the emitter barrier. Moreover, the large signal characterization of the HET is shown for different layouts, where the collector barrier was scaled.

Databases and Associated Tools for Glycomics and Glycoproteomics.

PubMed

Lisacek, Frederique; Mariethoz, Julien; Alocci, Davide; Rudd, Pauline M; Abrahams, Jodie L; Campbell, Matthew P; Packer, Nicolle H; Ståhle, Jonas; Widmalm, Göran; Mullen, Elaine; Adamczyk, Barbara; Rojas-Macias, Miguel A; Jin, Chunsheng; Karlsson, Niclas G

2017-01-01

The access to biodatabases for glycomics and glycoproteomics has proven to be essential for current glycobiological research. This chapter presents available databases that are devoted to different aspects of glycobioinformatics. This includes oligosaccharide sequence databases, experimental databases, 3D structure databases (of both glycans and glycorelated proteins) and association of glycans with tissue, disease, and proteins. Specific search protocols are also provided using tools associated with experimental databases for converting primary glycoanalytical data to glycan structural information. In particular, researchers using glycoanalysis methods by U/HPLC (GlycoBase), MS (GlycoWorkbench, UniCarb-DB, GlycoDigest), and NMR (CASPER) will benefit from this chapter. In addition we also include information on how to utilize glycan structural information to query databases that associate glycans with proteins (UniCarbKB) and with interactions with pathogens (SugarBind).

Chapter 51: How to Build a Simple Cone Search Service Using a Local Database

NASA Astrophysics Data System (ADS)

Kent, B. R.; Greene, G. R.

The cone search service protocol will be examined from the server side in this chapter. A simple cone search service will be setup and configured locally using MySQL. Data will be read into a table, and the Java JDBC will be used to connect to the database. Readers will understand the VO cone search specification and how to use it to query a database on their local systems and return an XML/VOTable file based on an input of RA/DEC coordinates and a search radius. The cone search in this example will be deployed as a Java servlet. The resulting cone search can be tested with a verification service. This basic setup can be used with other languages and relational databases.

Chapter 1 Historical Background on Gamete and Embryo Cryopreservation.

PubMed

Ali, Jaffar; AlHarbi, Naif H; Ali, Nafisa

2017-01-01

This chapter describes the development of the science of cryopreservation of gametes and embryos of various species including human. It attempts to record in brief the main contributions of workers in their attempts to cryopreserve gametes and embryos. The initial difficulties faced and subsequent developments and triumphs leading to present-day state of the art are given in a concise manner. The main players and their contributions are mentioned and the authors' aim is to do justice to them. This work also attempts to ensure that credit is correctly attributed for significant advances in gamete and embryo cryopreservation. In general this chapter has tried to describe the historical development of the science of cryopreservation of gametes and embryos as accurately as possible without bias or partiality.

A visual analytical approach to rock art panel condition assessment

NASA Astrophysics Data System (ADS)

Vogt, Brandon J.

Rock art is a term for pecked, scratched, or painted symbols found on rock surfaces, most typically joint faces called rock art panels. Because rock art exists on rock at the atmosphere interface, it is highly susceptible to the destructive processes of weathering. Thus, rock weathering scientists, including those that study both natural and cultural surfaces, play a key role towards understanding rock art longevity. The mapping of weathering forms on rock art panels serves as a basis from which to assess overall panel instability. This work examines fissures, case hardened surfaces, crumbly disintegration, and lichen. Knowledge of instability, as measured through these and other weathering forms, provides integral information to land managers and archaeological conservators required to prioritize panels for remedial action. The work is divided into five chapters, three of which are going to be submitted as a peer-reviewed journal manuscript. The second chapter, written as a manuscript for International Newsletter on Rock Art, describes a specific set of criteria that lead to the development of a mapping tool for weathering forms, called 'mapping weathering forms in three dimensions' (MapWeF). The third chapter, written as a manuscript for Remote Sensing of Environment, presents the methodology used to develop MapWeF. The chapter incorporates terrestrial laser scanning, a geographic information system (GIS), geovisualization, image analysis, and exploratory spatial data analysis (ESDA) to identify, map, and quantify weathering features known to cause instability on rock art panels. The methodology implemented in the third chapter satisfies the criteria described in Chapter Two. In the fourth chapter, prepared as a manuscript for Geomorphology, MapWeF is applied to a site management case study, focusing on a region---southeastern Colorado---with notoriously weak and endangered sandstone rock art panels. The final conclusions chapter describes contributions of the work to GIScience and rock weathering, and discusses how MapWeF, as a diagnostic tool, fits into a larger national vision by linking existing rock art stability characterizations to cultural resource management-related conservation action.

Patient-Specific Induced Pluripotent Stem Cell Models: Generation and Characterization of Cardiac Cells.

PubMed

Zanella, Fabian; Sheikh, Farah

2016-01-01

The generation of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes has been of utmost interest for the study of cardiac development, cardiac disease modeling, and evaluation of cardiotoxic effects of novel candidate drugs. Several protocols have been developed to guide human stem cells toward the cardiogenic path. Pioneering work used serum to promote cardiogenesis; however, low cardiogenic throughputs, lack of chemical definition, and batch-to-batch variability of serum lots constituted a considerable impediment to the implementation of those protocols to large-scale cell biology. Further work focused on the manipulation of pathways that mouse genetics indicated to be fundamental in cardiac development to promote cardiac differentiation in stem cells. Although extremely elegant, those serum-free protocols involved the use of human recombinant cytokines that tend to be quite costly and which can also be variable between lots. The latest generation of cardiogenic protocols aimed for a more cost-effective and reproducible definition of the conditions driving cardiac differentiation, using small molecules to manipulate cardiogenic pathways overriding the need for cytokines. This chapter details methods based on currently available cardiac differentiation protocols for the generation and characterization of robust numbers of hiPSC-derived cardiomyocytes under chemically defined conditions.

Cloning mice and ES cells by nuclear transfer from somatic stem cells and fully differentiated cells.

PubMed

Wang, Zhongde

2011-01-01

Cloning animals by nuclear transfer (NT) has been successful in several mammalian species. In addition to cloning live animals (reproductive cloning), this technique has also been used in several species to establish cloned embryonic stem (ntES) cell lines from somatic cells. It is the latter application of this technique that has been heralded as being the potential means to produce isogenic embryonic stem cells from patients for cell therapy (therapeutic cloning). These two types of cloning differ only in the steps after cloned embryos are produced: for reproductive cloning the cloned embryos are transferred to surrogate mothers to allow them to develop to full term and for therapeutic cloning the cloned embryos are used to derive ntES cells. In this chapter, a detailed NT protocol in mouse by using somatic stem cells (neuron and skin stem cells) and fully differentiated somatic cells (cumulus cells and fibroblast cells) as nuclear donors is described.

In vitro propagation of peanut (Arachis hypogaea L.) by shoot tip culture.

PubMed

Ozudogru, Elif Aylin; Kaya, Ergun; Lambardi, Maurizio

2013-01-01

Peanut (Arachis hypogaea L.), also known as groundnut, is the most important species of Arachis genus, originating from Brazil and Peru. Peanut seeds contain high seed oil, proteins, amino acids, and vitamin E, and are consumed worldwide as edible nut, peanut butter, or candy, and peanut oil extracted from the seeds. The meal remaining after oil extraction is also used for animal feed. However, its narrow germplasm base, together with susceptibility to diseases, pathogens, and weeds, decreases yield and seed quality and causes great economic losses annually. Hence, the optimization of efficient in vitro propagation procedures would be highly effective for peanut propagation, as it would raise yield and improve seed quality and flavor. Earlier reports on traditional micropropagation methods, based on axillary bud proliferation which guarantees the multiplication of true-to-type plants, are still limited. This chapter describes a micropropagation protocol to improve multiple shoot formation from shoot-tip explants by using AgNO(3) in combination with plant growth regulators.