Isolation of cancer cells by "in situ" microfluidic biofunctionalization protocols

KAUST Repository

De Vitis, Stefania; Matarise, Giuseppina; Pardeo, Francesca; Catalano, Rossella; Malara, Natalia Maria; Trunzo, Valentina; Tallerico, Rossana; Gentile, Francesco T.; Candeloro, Patrizio; Coluccio, Maria Laura; Massaro, Alessandro S.; Viglietto, Giuseppe; Carbone, Ennio; Kutter, Jö rg Peter; Perozziello, Gerardo; Di Fabrizio, Enzo M.

2014-01-01

The aim of this work is the development of a microfluidic immunosensor for the immobilization of cancer cells and their separation from healthy cells by using "in situ" microfluidic biofunctionalization protocols. These protocols allow to link antibodies on microfluidic device surfaces and can be used to study the interaction between cell membrane and biomolecules. Moreover they allow to perform analysis with high processing speed, small quantity of reagents and samples, short reaction times and low production costs. In this work the developed protocols were used in microfluidic devices for the isolation of cancer cells in heterogeneous blood samples by exploiting the binding of specific antibody to an adhesion protein (EpCAM), overexpressed on the tumor cell membranes. The presented biofunctionalization protocols can be performed right before running the experiment: this allows to have a flexible platform where biomolecules of interest can be linked on the device surface according to the user's needs. © 2014 Elsevier B.V. All rights reserved.

Isolation of cancer cells by "in situ" microfluidic biofunctionalization protocols

KAUST Repository

De Vitis, Stefania

2014-07-01

The aim of this work is the development of a microfluidic immunosensor for the immobilization of cancer cells and their separation from healthy cells by using "in situ" microfluidic biofunctionalization protocols. These protocols allow to link antibodies on microfluidic device surfaces and can be used to study the interaction between cell membrane and biomolecules. Moreover they allow to perform analysis with high processing speed, small quantity of reagents and samples, short reaction times and low production costs. In this work the developed protocols were used in microfluidic devices for the isolation of cancer cells in heterogeneous blood samples by exploiting the binding of specific antibody to an adhesion protein (EpCAM), overexpressed on the tumor cell membranes. The presented biofunctionalization protocols can be performed right before running the experiment: this allows to have a flexible platform where biomolecules of interest can be linked on the device surface according to the user\\'s needs. © 2014 Elsevier B.V. All rights reserved.

Comparison of in vitro and in situ methods in evaluation of forage digestibility in ruminants.

Science.gov (United States)

Krizsan, S J; Nyholm, L; Nousiainen, J; Südekum, K-H; Huhtanen, P

2012-09-01

The objective of this study was to compare the application of different in vitro and in situ methods in empirical and mechanistic predictions of in vivo OM digestibility (OMD) and their associations to near-infrared reflectance spectroscopy spectra for a variety of forages. Apparent in vivo OMD of silages made from alfalfa (n = 2), corn (n = 9), corn stover (n = 2), grass (n = 11), whole crops of wheat and barley (n = 8) and red clover (n = 7), and fresh alfalfa (n = 1), grass hays (n = 5), and wheat straws (n = 5) had previously been determined in sheep. Concentrations of indigestible NDF (iNDF) in all forage samples were determined by a 288-h ruminal in situ incubation. Gas production of isolated forage NDF was measured by in vitro incubations for 72 h. In vitro pepsin-cellulase OM solubility (OMS) of the forages was determined by a 2-step gravimetric digestion method. Samples were also subjected to a 2-step determination of in vitro OMD based on buffered rumen fluid and pepsin. Further, rumen fluid digestible OM was determined from a single 96-h incubation at 38°C. Digestibility of OM from the in situ and the in vitro incubations was calculated according to published empirical equations, which were either forage specific or general (1 equation for all forages) within method. Indigestible NDF was also used in a mechanistic model to predict OMD. Predictions of OMD were evaluated by residual analysis using the GLM procedure in SAS. In vitro OMS in a general prediction equation of OMD did not display a significant forage-type effect on the residuals (observed - predicted OMD; P = 0.10). Predictions of OMD within forage types were consistent between iNDF and the 2-step in vitro method based on rumen fluid. Root mean square error of OMD was least (0.032) when the prediction was based on a general forage equation of OMS. However, regenerating a simple regression for iNDF by omitting alfalfa and wheat straw reduced the root mean square error of OMD to 0

An improved in situ method for determining depth distributions of gamma-ray emitting radionuclides

International Nuclear Information System (INIS)

Benke, R.R.; Kearfott, K.J.

2001-01-01

In situ gamma-ray spectrometry determines the quantities of radionuclides in some medium with a portable detector. The main limitation of in situ gamma-ray spectrometry lies in determining the depth distribution of radionuclides. This limitation is addressed by developing an improved in situ method for determining the depth distributions of gamma-ray emitting radionuclides in large area sources. This paper implements a unique collimator design with conventional radiation detection equipment. Cylindrically symmetric collimators were fabricated to allow only those gamma-rays emitted from a selected range of polar angles (measured off the detector axis) to be detected. Positioned with its axis normal to surface of the media, each collimator enables the detection of gamma-rays emitted from a different range of polar angles and preferential depths. Previous in situ methods require a priori knowledge of the depth distribution shape. However, the absolute method presented in this paper determines the depth distribution as a histogram and does not rely on such assumptions. Other advantages over previous in situ methods are that this method only requires a single gamma-ray emission, provides more detailed depth information, and offers a superior ability for characterizing complex depth distributions. Collimated spectrometer measurements of buried area sources demonstrated the ability of the method to yield accurate depth information. Based on the results of actual measurements, this method increases the potential of in situ gamma-ray spectrometry as an independent characterization tool in situations with unknown radionuclide depth distributions

Development of analysis methods for seismically isolated nuclear structures

International Nuclear Information System (INIS)

Yoo, Bong; Lee, Jae-Han; Koo, Gyeng-Hoi

2002-01-01

KAERI's contributions to the project entitled Development of Analysis Methods for Seismically Isolated Nuclear Structures under IAEA CRP of the intercomparison of analysis methods for predicting the behaviour of seismically isolated nuclear structures during 1996-1999 in effort to develop the numerical analysis methods and to compare the analysis results with the benchmark test results of seismic isolation bearings and isolated nuclear structures provided by participating countries are briefly described. Certain progress in the analysis procedures for isolation bearings and isolated nuclear structures has been made throughout the IAEA CRPs and the analysis methods developed can be improved for future nuclear facility applications. (author)

Demonstration of a collimated in situ method for determining depth distributions using gamma-ray spectrometry

CERN Document Server

Benke, R R

2002-01-01

In situ gamma-ray spectrometry uses a portable detector to quantify radionuclides in materials. The main shortcoming of in situ gamma-ray spectrometry has been its inability to determine radionuclide depth distributions. Novel collimator designs were paired with a commercial in situ gamma-ray spectrometry system to overcome this limitation for large area sources. Positioned with their axes normal to the material surface, the cylindrically symmetric collimators limited the detection of un attenuated gamma-rays from a selected range of polar angles (measured off the detector axis). Although this approach does not alleviate the need for some knowledge of the gamma-ray attenuation characteristics of the materials being measured, the collimation method presented in this paper represents an absolute method that determines the depth distribution as a histogram, while other in situ methods require a priori knowledge of the depth distribution shape. Other advantages over previous in situ methods are that this method d...

Some Orchid Species Fungi Isolated by Different Methods

Directory of Open Access Journals (Sweden)

Arzu ÇIĞ

2014-03-01

Full Text Available Due to their very small seeds that do not contain endosperm, many terrestrial orchid species require the presence of fungi in order to germinate and maintain their lives; and symbiotic culture studies are being carried out on this topic. For the purpose of determining the orchid species on which the fungus to be used as inoculants in the symbiotic culture will be effective, fungi isolated through several isolation methods are cultured with orchid species. In this study a total of four different isolation methods were applied as one on the tubers and rhizomes and three on the soil of eleven orchid species from the Anacamptis, Cephalanthera, Dactylorhiza and Orchis genera. Three different culture media were used in the methods. At the end of the study Alternaria, Aspergillus, Fusarium, Macrophomina, Rhizoctonia, Trichoderma and Verticillium fungi were isolated. In the study that was conducted with the aimed to isolate particularly Rhizoctania spp. fungi, the fungi was isolated from the tubers of Dactylorhiza umbrosa and Orchis palustris species and the soil of the Orchis simia species. Fusarium and Aspergillus species were isolated the most in all implemented methods and from all species.

Radioactive waste isolation in salt: Peer review of the Golder Associates draft test plan for in situ testing in an exploratory shaft in salt

International Nuclear Information System (INIS)

Hambley, D.F.; Mraz, D.Z.; Unterberter, R.R.

1987-01-01

This report documents the peer review conducted by Argonne National Laboratory of a document entitled ''Draft Test Plan for In Situ Testing in an Exploratory Shaft in Salt,'' prepared for Battelle Memorial Institute's Office of Nuclear Waste Isolation by Golder Associates, Inc. In general, the peer review panelists found the test plan to be technically sound, although some deficiencies were identified. Recommendations for improving the test plan are presented in this review report. A microfiche copy of the following unpublished report is attached to the inside back cover of this report: ''Draft Test Plan for In Situ Testing in an Exploratory Shaft in Salt,'' prepared by Golder Associates, Inc., for Office of Nuclear Waste Isolation, Battelle Memorial Institute, Columbus, Ohio (March 1985)

In situ methods for Li-ion battery research: A review of recent developments

Science.gov (United States)

Harks, P. P. R. M. L.; Mulder, F. M.; Notten, P. H. L.

2015-08-01

A considerable amount of research is being directed towards improving lithium-ion batteries in order to meet today's market demands. In particular in situ investigations of Li-ion batteries have proven extremely insightful, but require the electrochemical cell to be fully compatible with the conditions of the testing method and are therefore often challenging to execute. Advantageously, in the past few years significant progress has been made with new, more advanced, in situ techniques. Herein, a comprehensive overview of in situ methods for studying Li-ion batteries is given, with the emphasis on new developments and reported experimental highlights.

K. OXYTOCA BACTERIOPHAGES ISOLATION METHODS IMPROVEMENT

Directory of Open Access Journals (Sweden)

G. R. Sadrtdinova

2017-01-01

Full Text Available The article presents the results of a study related to increasing the efficiency of phage isolation of bacteria of the species K. oxytoca, by developing the optimal composition of the medium used in the work. In scientific research, in almost all methods associated with the isolation of bacteriophages, meat-peptone broth and meat-peptone agar are used as the nutrient basis. The peculiarities of growth and cultivation of microorganisms create certain difficulties for the isolation of phages active against bacteria of the species K. oxytoca. The selection of components and the creation of an environment that would ensure the optimal growth of both the bacterial culture and the reproduction of the virus makes it possible to facilitate the isolation of bacteriophages. The number of bacterial strains used in the work was 7. All strains of cultures were obtained from the Museum of the Department of Microbiology, Virology, Epizootology and Veterinary and Sanitary Expertise of the Federal State Budget Educational Institution of Higher Education “Ulyanovsk State Agrarian University named after P.A. Stolypin”. The studies included 2 main stages. The first stage consisted in isolation of bacteriophages by the method of isolation from the external environment by the method of Adelson L.I., Lyashenko E.A. The material for the studies were samples: soil, sewage sample, fecal samples (2. Only 4 samples. According to the chosen method, the sowing of the putative phagolysate was carried out on meat-peptone agar (1.5% and the agar for isolating bacteriophages (Aph (1.5%. A positive result was the presence on the environment of negative colonies, clearly visible on the matt background of deep growth of bacteria. A negative result is a continuous growth (“lawn” of bacterial culture. As a control, the culture of the microorganism studied was used for the media. In the course of the conducted studies for the first stage, 2 bacteriophages were isolated, active

Highly Dense Isolated Metal Atom Catalytic Sites

DEFF Research Database (Denmark)

Chen, Yaxin; Kasama, Takeshi; Huang, Zhiwei

2015-01-01

-ray diffraction. A combination of electron microscopy images with X-ray absorption spectra demonstrated that the silver atoms were anchored on five-fold oxygen-terminated cavities on the surface of the support to form highly dense isolated metal active sites, leading to excellent reactivity in catalytic oxidation......Atomically dispersed noble-metal catalysts with highly dense active sites are promising materials with which to maximise metal efficiency and to enhance catalytic performance; however, their fabrication remains challenging because metal atoms are prone to sintering, especially at a high metal...... loading. A dynamic process of formation of isolated metal atom catalytic sites on the surface of the support, which was achieved starting from silver nanoparticles by using a thermal surface-mediated diffusion method, was observed directly by using in situ electron microscopy and in situ synchrotron X...

Laboratory-performance criteria for in situ waste-stabilization materials

International Nuclear Information System (INIS)

Shaw, P.; Weidner, J.

1996-01-01

The Department of Energy (DOE) Landfill Stabilization Focus Area is investigating a variety of in situ placement methods, grout materials, and characterization techniques for the stabilization of buried low-level transuranic-contaminated waste at Department of Energy sites. In situ stabilization involves underground injection or placement of substances to isolate, treat, or contain buried contaminants. Performance criteria were developed to evaluate various candidate stabilization materials for both long-term stabilization and interim stabilization or retrieval. The criteria are go/no-go, ready, and preliminary. The criterion go/no-go eliminates technologies that are not applicable for in situ treatment of buried waste. The criterion ready indicates that the technology is sufficiently developed and proven to be field demonstrated full-scale. The criterion preliminary indicates the prospective technologies to be potentially applicable to in situ buried waste stabilization, but further development is needed before the technology is ready for field-scale demonstration

Comparative study of in-situ filter test methods

International Nuclear Information System (INIS)

Marshall, M.; Stevens, D.C.

1981-01-01

Available methods of testing high efficiency particulate aerosol (HEPA) filters in-situ have been reviewed. In order to understand the relationship between the results produced by different methods a selection has been compared. Various pieces of equipment for generating and detecting aerosols have been tested and their suitability assessed. Condensation-nuclei, DOP (di-octyl phthalate) and sodium-flame in-situ filter test methods have been studied, using the 500 cfm (9000 m 3 /h) filter test rig at Harwell and in the field. Both the sodium-flame and DOP methods measure the penetration through leaks and filter material. However the measured penetration through filtered leaks depends on the aerosol size distribution and the detection method. Condensation-nuclei test methods can only be used to measure unfiltered leaks since condensation nuclei have a very low penetration through filtered leaks. A combination of methods would enable filtered and unfiltered leaks to be measured. A condensation-nucleus counter using n-butyl alcohol as the working fluid has the advantage of being able to detect any particle up to 1 μm in diameter, including DOP, and so could be used for this purpose. A single-particle counter has not been satisfactory because of interference from particles leaking into systems under extract, particularly downstream of filters, and because the concentration of the input aerosol has to be severely limited. The sodium-flame method requires a skilled operator and may cause safety and corrosion problems. The DOP method using a total light scattering detector has so far been the most satisfactory. It is fairly easy to use, measures reasonably low values of penetration and gives rapid results. DOP has had no adverse effect on HEPA filters over a long series of tests

A Modified Method for Isolation of Rhein from Senna

Science.gov (United States)

Mehta, Namita; Laddha, K. S.

2009-01-01

A simple and efficient method for the isolation of rhein from Cassia angustifolia (senna) leaves is described in which the hydrolysis of the sennosides and extraction of the hydrolysis products (free anthraquinones) is carried out in one step. Further isolation of rhein is achieved from the anthraquinone mixture. This method reduces the number of steps required for isolation of rhein as compared to conventional methods. PMID:20336207

Methods to isolate extracellular vesicles for diagnosis

Science.gov (United States)

Kang, Hyejin; Kim, Jiyoon; Park, Jaesung

2017-12-01

Extracellular vesicles (EVs) are small membrane-bound bodies that are released into extracellular space by diverse cells, and are found in body fluids like blood, urine and saliva. EVs contain RNA, DNA and proteins, which can be biomarkers for diagnosis. EVs can be obtained by minimally-invasive biopsy, so they are useful in disease diagnosis. High yield and purity contribute to precise diagnosis of disease, but damaged EVs and impurities can cause confu sed results. However, EV isolation methods have different yields and purities. Furthermore, the isolation method that is most suitable to maximize EV recovery efficiency depends on the experimental conditions. This review focuses on merits and demerits of several types of EV isolation methods, and provides examples of how to diagnose disease by exploiting information obtained by analysis of EVs.

Method for in situ carbon deposition measurement for solid oxide fuel cells

Science.gov (United States)

Kuhn, J.; Kesler, O.

2014-01-01

Previous methods to measure carbon deposition in solid oxide fuel cell (SOFC) anodes do not permit simultaneous electrochemical measurements. Electrochemical measurements supplemented with carbon deposition quantities create the opportunity to further understand how carbon affects SOFC performance and electrochemical impedance spectra (EIS). In this work, a method for measuring carbon in situ, named here as the quantification of gasified carbon (QGC), was developed. TGA experiments showed that carbon with a 100 h residence time in the SOFC was >99.8% gasified. Comparison of carbon mass measurements between the TGA and QGC show good agreement. In situ measurements of carbon deposition in SOFCs at varying molar steam/carbon ratios were performed to further validate the QGC method, and suppression of carbon deposition with increasing steam concentration was observed, in agreement with previous studies. The technique can be used to investigate in situ carbon deposition and gasification behavior simultaneously with electrochemical measurements for a variety of fuels and operating conditions, such as determining conditions under which incipient carbon deposition is reversible.

Study on design method for seismically isolated FBR plants

International Nuclear Information System (INIS)

Hirata, Kazuta; Yabana, Shuichi; Ohtori, Yasuki; Ishida, Katsuhiko; Sawada, Yoshihiro; Shiojiri; Hiroo; Mazda, Taiji

1998-01-01

CRIEPI conducted 'Demonstration test on FBR seismic isolation system' from 1987 to 1996 under contract with Ministry of International Trade and Industry, Japan. In the demonstration test, base isolation technologies are prepared and demonstrated to apply to FBR and the design guidelines are proposed. In this report overall contents of the design guidelines entitled Design guidelines for seismically base isolated FBR plants' are included. The design guidelines, as a rule, are limited to apply to FBR plants where entire reactor building is isolated in the horizontal direction using laminated rubber bearings as isolators. The design guidelines and its concepts, however, will be useful for the development of similar guidelines for other isolation systems using different type of isolation methods and other nuclear facilities. The design guidelines consist of three parts and appendices. The first part is 'Policy for Safety Design of Base Isolated FBR Plants' specifying the principles and the requirements in the planning and the design for the safety of base isolated FBR plants. The second part is Policy for Seismic Design of Base Isolated FBR' describing the principles and the requirements in the seismic design and the evaluation of safety for base isolated FBR plants. The third part is 'Design Methods for Seismic Isolated FBR Plants' detailing the methods, procedures and parameters to be used in the design and the evaluation of safety fro base isolated FBR plants. In appendices examples of design procedures for base isolated reactor building and laminated rubber bearings as well as various test data on laminated rubber bearings, etc. are shown. (author)

In situ production of bio-surfactants: An alternative method for dispersing and bioremediating marine oil spills

International Nuclear Information System (INIS)

Josefsen, K.D.; Sveum, P.; Ramstad, P.; Markussen, S.; Folkvord, K.; Krigsvoll, K.; Aune, R.; Storroe, I.

1995-01-01

Some oil degrading bacteria are able to produce surfactants. These biosurfactants enhance dispersion of oil droplets into the water column. A large number of surfactant producing bacterial strains have been isolated from seawater samples collected at different sites around the world. Strains isolated from seawater samples collected in cold regions generally had better properties than strains isolated from warm seawater. Many of the isolated strains were able to disperse crude oils with a large variation of composition, as well as the water-in-emulsion (chocolate mousse) formed during weathering of crude oil in the sea. The results show that in situ application of surfactant producing bacteria can be a viable tool in future oil spill contingency, and that dispersion of oil may increase the biodegradation rate. Work is in progress to examine the use of such bacteria in the bioremediation of oil contaminated shorelines. 10 refs., 3 figs., 2 tabs

The potential of high resolution ultrasonic in-situ methods

International Nuclear Information System (INIS)

Schuster, K.

2010-01-01

source and receivers and on the applied frequencies. With the help of identified phases in the ultrasonic wave field, their travel times and appropriate amplitudes kinematic and dynamic parameters are derived routinely. The most important are: P-wave velocity (longitudinal or compressional wave velocity (vp)), Sv-wave velocity (vertical polarised transversal or shear wave velocity (vsv), absolute amplitudes of first arrival (P-wave) and Sv-wave onset phases, normalised amplitudes of first arrival (P-wave) and Sv-wave onset phases, apparent frequency of first arrival phases (P-wave) and Sv-wave onset phases, in-situ dynamic elastic Poisson's ratio, in-situ dynamic elastic Young's modulus and in-situ dynamic elastic modulus of rigidity. The main qualities which were detected and/or characterised with the help of these parameters in the past years are: - Degree and extent of EDZ/EdZ. - Long and short term development of EDZ/EdZ (application of repetition measurements). - Borehole disturbed Zones (BdZ). - Small scale rock heterogeneities. - Seismic anisotropy. - Stress orientation via rotational interval velocity measurements. - Determination of dynamic elastic in-situ parameters. Results from repetition measurements showed the high reliability of the methods. Furthermore, in several experiments the ultrasonic methods were combined with other geotechnical or geophysical methods, for example permeability and geo-electrical methods. Results from a seismic anisotropy study at the Mont Terri Rock Laboratory are shown. Two sub-horizontal 2 m long boreholes with a distance of 1 m were used. Nearly 600 different ray paths were analysed. Depending on the angles of the ray paths (60 deg. -300 deg. and 120 deg. -240 deg. ) the P-wave velocities vary between 2600 and 3100 m/s. Lower velocities are supposed to be influenced by the EDZ. (authors)

Quantitative comparison of in situ soil CO2 flux measurement methods

Science.gov (United States)

Jennifer D. Knoepp; James M. Vose

2002-01-01

Development of reliable regional or global carbon budgets requires accurate measurement of soil CO2 flux. We conducted laboratory and field studies to determine the accuracy and comparability of methods commonly used to measure in situ soil CO2 fluxes. Methods compared included CO2...

Estimating grass and grass silage degradation characteristics by in situ and in vitro gas production methods

Directory of Open Access Journals (Sweden)

Danijel Karolyi

2010-01-01

Full Text Available Fermentation characteristics of grass and grass silage at different maturities were studied using in situ and in vitro gas production methods. In situ data determined difference between grass and silage. Degradable fraction decreased as grass matured while the undegradable fraction increased. Rate of degradation (kd was slower for silage than fresh grass. Gas production method (GP data showed that fermentation of degradable fraction was different between stage of maturity in both grass and silage. Other data did not show any difference with the exception for the rate of GP of soluble and undegradable fraction. The in situ degradation characteristics were estimated from GP characteristics. The degradable and undegradable fractions could be estimated by multiple relationships. Using the three-phases model for gas production kd and fermentable organic matter could be estimated from the same parameters. The only in situ parameter that could not be estimated with GP parameters was the soluble fraction. The GP method and the three phases model provided to be an alternative to the in situ method for animal feed evaluations.

Improved recovery of DNA from polyacrylamide gels after in situ DNA footprinting

NARCIS (Netherlands)

van Keulen, G; Meijer, WG

Methods used to date for the isolation of DNA from polyacrylamide gels are elution based, time-consuming and with low yield in DNA. This paper describes an improved system employing polyacrylamide gels made of a meltable matrix. The new system was successfully applied to in situ DNA footprinting

In situ feeding rates of plantonic copepods: A comparison of four methods

DEFF Research Database (Denmark)

Kiørboe, Thomas; Møhlenberg, Flemming; Riisgård, Hans Ulrik

1985-01-01

into estimates of in situ algal grazing rates by means of independently estimated gut turnover times, and were compared with chlorophyll and particle-volume grazing rates of animals sampled simultaneously and incubated in water from the collection depth. In addition, egg-production rates of adult females were...... problems of the different methods are discussed, and it is concluded that they all approach representative (although minimum) estimates of in situ feeding rates....

A new rapid method for isolating nucleoli.

Science.gov (United States)

Li, Zhou Fang; Lam, Yun Wah

2015-01-01

The nucleolus was one of the first subcellular organelles to be isolated from the cell. The advent of modern proteomic techniques has resulted in the identification of thousands of proteins in this organelle, and live cell imaging technology has allowed the study of the dynamics of these proteins. However, the limitations of current nucleolar isolation methods hinder the further exploration of this structure. In particular, these methods require the use of a large number of cells and tedious procedures. In this chapter we describe a new and improved nucleolar isolation method for cultured adherent cells. In this method cells are snap-frozen before direct sonication and centrifugation onto a sucrose cushion. The nucleoli can be obtained within a time as short as 20 min, and the high yield allows the use of less starting material. As a result, this method can capture rapid biochemical changes in nucleoli by freezing the cells at a precise time, hence faithfully reflecting the protein composition of nucleoli at the specified time point. This protocol will be useful for proteomic studies of dynamic events in the nucleolus and for better understanding of the biology of mammalian cells.

In situ thermal properties characterization using frequential methods

Energy Technology Data Exchange (ETDEWEB)

Carpentier, O.; Defer, D.; Antczak, E.; Chauchois, A.; Duthoit, B. [Laboratoire dArtois de Mecanique Thermique Instrumentation (LAMTI), FSA Universite dArtois, Technoparc Futura, 62400 Bethune (France)

2008-07-01

In numerous fields, especially that of geothermal energy, we need to know about the thermal behaviour of the soil now that the monitoring of renewable forms of energy is an ecological, economic and scientific issue. Thus heat from the soil is widely used for air-conditioning systems in buildings both in Canada and in the Scandinavian countries, and it is spreading. The effectiveness of this technique is based on the soils calorific potential and its thermophysical properties which will define the quality of the exchanges between the soil and a heat transfer fluid. This article puts forward a method to be used for the in situ thermophysical characterisation of a soil. It is based upon measuring the heat exchanges on the surface of the soil and on measuring a temperature a few centimetres below the surface. The system is light, inexpensive, well-suited to the taking of measurements in situ without the sensors used introducing any disturbance into the heat exchanges. Whereas the majority of methods require excitation, the one presented here is passive and exploits natural signals. Based upon a few hours of recording, the natural signals allow us to identify the soils thermophysical properties continuously. The identification is based upon frequency methods the quality of which can be seen when the thermophysical properties are injected into a model with finite elements by means of a comparison of the temperatures modelled and those actually measured on site. (author)

In-situ spectrometry of 137Cs in the soil by unfolding method

International Nuclear Information System (INIS)

Fueloep, M.; Ragan, P.; Krnac, S.

1995-01-01

This contribution is aimed to the possibility of improving the in-situ gamma spectrometry to be independent on a knowledge about a depth distribution of 137 Cs in soil and sufficiently sensitive for the measurement of the post-Chernobyl 137 Cs at present, as well. The depth distribution of 137 Cs averaged over a large area of soil is obtained by unfolding of the detector responses to primary and in soil forward scattered photons. The proposed method employs detector with and without collimator. The 137 Cs distributions obtained in-situ measurements are analysed, and comparisons are made to the results obtained with soil sampling and with standard in-situ spectrometry, as well. 5 figs., 1 tab., 4 refs

A magnetic micropore chip for rapid (<1 hour) unbiased circulating tumor cell isolation and in situ RNA analysis.

Science.gov (United States)

Ko, Jina; Bhagwat, Neha; Yee, Stephanie S; Black, Taylor; Redlinger, Colleen; Romeo, Janae; O'Hara, Mark; Raj, Arjun; Carpenter, Erica L; Stanger, Ben Z; Issadore, David

2017-09-12

The use of microtechnology for the highly selective isolation and sensitive detection of circulating tumor cells has shown enormous promise. One challenge for this technology is that the small feature sizes - which are the key to this technology's performance - can result in low sample throughput and susceptibility to clogging. Additionally, conventional molecular analysis of CTCs often requires cells to be taken off-chip for sample preparation and purification before analysis, leading to the loss of rare cells. To address these challenges, we have developed a microchip platform that combines fast, magnetic micropore based negative immunomagnetic selection (>10 mL h -1 ) with rapid on-chip in situ RNA profiling (>100× faster than conventional RNA labeling). This integrated chip can isolate both rare circulating cells and cell clusters directly from whole blood and allow individual cells to be profiled for multiple RNA cancer biomarkers, achieving sample-to-answer in less than 1 hour for 10 mL of whole blood. To demonstrate the power of this approach, we applied our device to the circulating tumor cell based diagnosis of pancreatic cancer. We used a genetically engineered lineage-labeled mouse model of pancreatic cancer (KPCY) to validate the performance of our chip. We show that in a cohort of patient samples (N = 25) that this device can detect and perform in situ RNA analysis on circulating tumor cells in patients with pancreatic cancer, even in those with extremely sparse CTCs (<1 CTC mL -1 of whole blood).

ADVANTAGES/DISADVANTAGES FOR ISCO METHODS IN-SITU FENTON OXIDATION IN-SITU PERMANGANATE OXIDATION

Science.gov (United States)

The advantages and disadvantages of in-situ Fenton oxidation and in-situ permanganate oxidation will be presented. This presentation will provide a brief overview of each technology and a detailed analysis of the advantages and disadvantages of each technology. Included in the ...

Rapid and inexpensive method for isolating plasmid DNA

International Nuclear Information System (INIS)

Aljanabi, S. M.; Al-Awadi, S. J.; Al-Kazaz, A. A.; Baghdad Univ.

1997-01-01

A small-scale and economical method for isolating plasmid DNA from bacteria is described. The method provides DNA of suitable quality for most DNA manipulation techniques. This DNA can be used for restriction endonuclease digestion, southern blot hybridization, nick translation and end labeling of DNA probes, Polymerase Chain Reaction (PCR) -based techniques, transformation, DNA cycle-sequencing, and Chain-termination method for DNA sequencing. The entire procedure is adapted to 1.5 ml microfuge tubes and takes approximately 30 mins. The DNA isolated by this method has the same purity produced by CTAB and cesium chloride precipitation and purification procedures respectively. The two previous methods require many hours to obtain the final product and require the use of very expensive equipment as ultracentrifuge. This method is well suited for the isolation of plasmid DNA from a large number of bacterial samples and in a very short time and low cost in laboratories where chemicals, expensive equipment and finance are limited factors in conducting molecular research. (authors). 11refs. 11refs

Methods and systems for in-situ electroplating of electrodes

Science.gov (United States)

Zappi, Guillermo Daniel; Zarnoch, Kenneth Paul; Huntley, Christian Andrew; Swalla, Dana Ray

2015-06-02

The present techniques provide electrochemical devices having enhanced electrodes with surfaces that facilitate operation, such as by formation of a porous nickel layer on an operative surface, particularly of the cathode. The porous metal layer increases the surface area of the electrode, which may result in increasing the efficiency of the electrochemical devices. The formation of the porous metal layer is performed in situ, that is, after the assembly of the electrodes into an electrochemical device. The in situ process offers a number of advantages, including the ability to protect the porous metal layer on the electrode surface from damage during assembly of the electrochemical device. The enhanced electrode and the method for its processing may be used in any number of electrochemical devices, and is particularly well suited for electrodes in an electrolyzer useful for splitting water into hydrogen and oxygen.

Signal processing methods for in-situ creep specimen monitoring

Science.gov (United States)

Guers, Manton J.; Tittmann, Bernhard R.

2018-04-01

Previous work investigated using guided waves for monitoring creep deformation during accelerated life testing. The basic objective was to relate observed changes in the time-of-flight to changes in the environmental temperature and specimen gage length. The work presented in this paper investigated several signal processing strategies for possible application in the in-situ monitoring system. Signal processing methods for both group velocity (wave-packet envelope) and phase velocity (peak tracking) time-of-flight were considered. Although the Analytic Envelope found via the Hilbert transform is commonly applied for group velocity measurements, erratic behavior in the indicated time-of-flight was observed when this technique was applied to the in-situ data. The peak tracking strategies tested had generally linear trends, and tracking local minima in the raw waveform ultimately showed the most consistent results.

Thermally Stable TiO2 - and SiO2 -Shell-Isolated Au Nanoparticles for In Situ Plasmon-Enhanced Raman Spectroscopy of Hydrogenation Catalysts.

Science.gov (United States)

Hartman, Thomas; Weckhuysen, Bert M

2018-03-12

Raman spectroscopy is known as a powerful technique for solid catalyst characterization as it provides vibrational fingerprints of (metal) oxides, reactants, and products. It can even become a strong surface-sensitive technique by implementing shell-isolated surface-enhanced Raman spectroscopy (SHINERS). Au@TiO 2 and Au@SiO 2 shell-isolated nanoparticles (SHINs) of various sizes were therefore prepared for the purpose of studying heterogeneous catalysis and the effect of metal oxide coating. Both SiO 2 - and TiO 2 -SHINs are effective SHINERS substrates and thermally stable up to 400 °C. Nano-sized Ru and Rh hydrogenation catalysts were assembled over the SHINs by wet impregnation of aqueous RuCl 3 and RhCl 3 . The substrates were implemented to study CO adsorption and hydrogenation under in situ conditions at various temperatures to illustrate the differences between catalysts and shell materials with SHINERS. This work demonstrates the potential of SHINS for in situ characterization studies in a wide range of catalytic reactions. © 2018 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

Static inflation attenuates ischemia/reperfusion injury in an isolated rat lung in situ.

Science.gov (United States)

Kao, Shang Jyh; Wang, David; Yeh, Diana Yu-Wung; Hsu, Kang; Hsu, Yung Hsiang; Chen, Hsing I

2004-08-01

Ischemia (I)/reperfusion (R) lung injury is an important clinical issue in lung transplantation. In the present study, we observed the effects of lung static inflation, different perfusates, and ventilatory gas with nitrogen or oxygen on the I/R-induced pulmonary damage. A total of 96 male Sprague-Dawley rats were used. The lung was isolated in situ. In an isolated lung, the capillary filtration coefficient (Kfc), lung weight gain (LWG), lung weight (LW)/body weight (BW) ratio, and protein concentration in BAL fluid (PCBAL) were measured or calculated to evaluate the degree of lung injury. Histologic examinations with hematoxylin-eosin staining were performed. I/R caused lung injury, as reflected by increases in Kfc, LWG, LW/BW, and PCBAL. The histopathologic picture revealed the presence of hyaline membrane formation and the infiltration of inflammatory cells. These values were significantly attenuated by static lung inflation. The I/R lung damage appeared to be less in the lung perfused with whole blood than in the lung perfused with an isotonic solution. Therapy with ventilatory air (ie, nitrogen or oxygen) did not alter the I/R lung damage. The data suggest that lung inflation is protective to I/R injury, irrespective of the type of ventilatory air used for treatment. The preservation of the lung for transplantation is better kept at a static inflation state and perfused with whole blood instead of an isotonic physiologic solution.

Nuclear methods for 'in-situ' geophysical investigations. 2

International Nuclear Information System (INIS)

Russell, J.P.; Oliver, D.W.

1985-01-01

There has been considerable effort expended by governmental, institutional, and private agencies in the development of suitable materials and instruments for application of nuclear methods for exploration and evaluation of mineral, oil, and gas resources. As a consequence of the peculiar circumstances of in situ investigations, much of the apparatus is, of design and construction, specifically adapted to the environment of the borehole (limited lateral dimension, high temperature and pressure, deep boreholes, etc.) which differs considerably from laboratory conditions. Meeting the needs for reliability and stability under adverse conditions of operation has led to often unique and innovative instrumentation. Much of the development effort has been provided by commercial service organizations whose interests require many details be maintained as proprietary. In addition, the tailoring of services to meet varying environmental and informational constraints has led to a wide range of instrument designs. As a result, it is difficult to provide an exhaustive description of all apparatus available for in situ investigations, rather, in this instance, systems and subsystems will be generically discussed. Radiation sources, detectors, and borehole probe designs are covered. (Auth.)

How the RNA isolation method can affect microRNA microarray results

DEFF Research Database (Denmark)

Podolska, Agnieszka; Kaczkowski, Bogumil; Litman, Thomas

2011-01-01

RNA microarray analysis on porcine brain tissue. One method is a phenol-guanidine isothiocyanate-based procedure that permits isolation of total RNA. The second method, miRVana™ microRNA isolation, is column based and recovers the small RNA fraction alone. We found that microarray analyses give different results...... that depend on the RNA fraction used, in particular because some microRNAs appear very sensitive to the RNA isolation method. We conclude that precautions need to be taken when comparing microarray studies based on RNA isolated with different methods.......The quality of RNA is crucial in gene expression experiments. RNA degradation interferes in the measurement of gene expression, and in this context, microRNA quantification can lead to an incorrect estimation. In the present study, two different RNA isolation methods were used to perform micro...

In situ rheology of yeast biofilms.

Science.gov (United States)

Brugnoni, Lorena I; Tarifa, María C; Lozano, Jorge E; Genovese, Diego

2014-01-01

The aim of the present work was to investigate the in situ rheological behavior of yeast biofilms growing on stainless steel under static and turbulent flow. The species used (Rhodototula mucilaginosa, Candida krusei, Candida kefyr and Candida tropicalis) were isolated from a clarified apple juice industry. The flow conditions impacted biofilm composition over time, with a predominance of C. krusei under static and turbulent flow. Likewise, structural variations occurred, with a tighter appearance under dynamic flow. Under turbulent flow there was an increase of 112 μm in biofilm thickness at 11 weeks (p < 0.001) and cell morphology was governed by hyphal structures and rounded cells. Using the in situ growth method introduced here, yeast biofilms were determined to be viscoelastic materials with a predominantly solid-like behavior, and neither this nor the G'0 values were significantly affected by the flow conditions or the growth time, and at large deformations their weak structure collapsed beyond a critical strain of about 1.5-5%. The present work could represent a starting point for developing in situ measurements of yeast rheology and contribute to a thin body of knowledge about fungal biofilm formation.

A molecular method for typing Herpes simplex virus isolates as an alternative to immunofluorescence methods

Directory of Open Access Journals (Sweden)

Abraham A

2009-01-01

Full Text Available Background: Typing of Herpes simplex virus (HSV isolates is required to identify the virus isolated in culture. The methods available for this include antigen detection by immunofluorescence (IF assays and polymerase chain reaction (PCR. This study was undertaken to standardize a molecular method for typing of HSV and compare it with a commercial IF reagent for typing. Objectives: To compare a molecular method for typing HSV isolates with a monoclonal antibody (MAb based IF test. Study design : This cross-sectional study utilized four reference strains and 42 HSV isolates obtained from patients between September 1998 and September 2004. These were subjected to testing using an MAb-based IF test and a PCR that detects the polymerase ( pol gene of HSV isolates. Results: The observed agreement of the MAb IF assay with the pol PCR was 95.7%. Fifty four point eight percent (23/42 of isolates tested by IF typing were found to be HSV-1, 40.5% (17/42 were HSV-2, and two (4.8% were untypable using the MAb IF assay. The two untypable isolates were found to be HSV-2 using the pol PCR. In addition, the cost per PCR test for typing is estimated to be around Rs 1,300 (USD 30, whereas the cost per MAb IF test is about Rs 1,500 (USD 35 including all overheads (reagents, instruments, personnel time, and consumables. Conclusion: The pol PCR is a cheaper and more easily reproducible method for typing HSV isolates as compared to the IF test. It could replace the IF-based method for routine typing of HSV isolates as availability of PCR machines (thermal cyclers is now more widespread than fluorescence microscopes in a country like India.

Effect of starch isolation method on properties of sweet potato starch

Directory of Open Access Journals (Sweden)

A. SURENDRA BABU

2014-08-01

Full Text Available Isolation method of starch with different agents influences starch properties, which provide attention for studying the most appropriate method for isolation of starch. In the present study sweet potato starch was isolated by Sodium metabisulphate (M1, Sodium chloride (M2, and Distilled water (M3 methods and these were assessed for functional, chemical, pasting and structural properties. M3 yielded the greatest recovery of starch (10.20%. Isolation methods significantly changed swelling power and pasting properties but starches exhibited similar chemical properties. Sweet potato starches possessed C-type diffraction pattern. Small size granules of 2.90 μm were noticed in SEM of M3 starch. A high degree positive correlation was found between ash, amylose, and total starch content. The study concluded that isolation methods brought changes in yield, pasting and structural properties of sweet potato starch.

Evaluation and selection of in-situ leaching mining method using analytic hierarchy process

International Nuclear Information System (INIS)

Zhao Heyong; Tan Kaixuan; Liu Huizhen

2007-01-01

According to the complicated conditions and main influence factors of in-situ leaching min- ing, a model and processes of analytic hierarchy are established for evaluation and selection of in-situ leaching mining methods based on analytic hierarchy process. Taking a uranium mine in Xinjiang of China for example, the application of this model is presented. The results of analyses and calculation indicate that the acid leaching is the optimum project. (authors)

Method for strontium isolation from high-mineralized water

International Nuclear Information System (INIS)

Evzhanov, Kh.; Andriyasova, G.M.

1983-01-01

A method to isolate strontium from high-mineralized waters containing sodium, magnesium, calcium and strontium chlorides, which differ from the prototype method in a considerable decrease in energy consumption with the preservation of a high degree of Sr, Mg and Ca isolation selectivity, has been suggested. According to the method suggested mineralized waters are treated with alkali (NaOH) in the amount of 95-97% of stoichiometry by magnesium, then after separation of magnesium hydroxide precipitate mother liquor is treated with sodium carbonate in the amount of 50-60% of stoichiometry by calcium. After separation of calcium carbonate precipitate mother liquor is treated with NaOH in the amount of 130-135% of stoichiometry by calcium. After separation of calcium hydroxide precipitate from mother liquor by means of sodium carbonate introduction strontium carbonate is isolated. The degree of strontium extraction in the form of SrCO 3 constitutes 90.5% of its content in the initial solution. The method presented can be used for strontium separation from natural and waste waters

A method to provide rapid in situ determination of tip radius in dynamic atomic force microscopy

International Nuclear Information System (INIS)

Santos, Sergio; Guang Li; Souier, Tewfik; Gadelrab, Karim; Chiesa, Matteo; Thomson, Neil H.

2012-01-01

We provide a method to characterize the tip radius of an atomic force microscopy in situ by monitoring the dynamics of the cantilever in ambient conditions. The key concept is that the value of free amplitude for which transitions from the attractive to repulsive force regimes are observed, strongly depends on the curvature of the tip. In practice, the smaller the value of free amplitude required to observe a transition, the sharper the tip. This general behavior is remarkably independent of the properties of the sample and cantilever characteristics and shows the strong dependence of the transitions on the tip radius. The main advantage of this method is rapid in situ characterization. Rapid in situ characterization enables one to continuously monitor the tip size during experiments. Further, we show how to reproducibly shape the tip from a given initial size to any chosen larger size. This approach combined with the in situ tip size monitoring enables quantitative comparison of materials measurements between samples. These methods are set to allow quantitative data acquisition and make direct data comparison readily available in the community.

In-situ spectrometry of {sup 137}Cs in the soil by unfolding method

Energy Technology Data Exchange (ETDEWEB)

Fueloep, M; Ragan, P [Inst. of Preventive and Clinical Medicine, 833301 Bratislava (Slovakia); Krnac, S [Slovak Technical Univ., Bratislava (Slovakia)

1996-12-31

This contribution is aimed to the possibility of improving the in-situ gamma spectrometry to be independent on a knowledge about a depth distribution of {sup 137}Cs in soil and sufficiently sensitive for the measurement of the post-Chernobyl {sup 137}Cs at present, as well. The depth distribution of {sup 137}Cs averaged over a large area of soil is obtained by unfolding of the detector responses to primary and in soil forward scattered photons. The proposed method employs detector with and without collimator. The {sup 137}Cs distributions obtained in-situ measurements are analysed, and comparisons are made to the results obtained with soil sampling and with standard in-situ spectrometry, as well. 5 figs., 1 tab., 4 refs.

Rock mechanics methods and in situ heater tests for design of a nuclear waste repository in basalt

International Nuclear Information System (INIS)

Board, M.P.

1978-01-01

Methods of integrating data from the Near-Surface Test Facility into the overall Waste Isolation Program are examined. Discussions are presented dealing primarily with the application of numerical models to the design of a waste repository. The various types of models currently available are discussed with reference to design in basalt and the breakdown of the problem of repository design is summarized. It is shown that the most efficient method for analyzing repository design is to break the problem down into several problems which are based on physical scale. These include the area directly surrounding a single waste canister (the very near field), the area including many canisters and canister emplacement rooms (the near field), and the area including the entire repository and the rock mass to the free surface (the far field). The methods by which numerical models are used for design are discussed. Flow charts are used to show the basic input data required, the calculational processes used, and the preliminary criteria for judgment of suitable repository performance. It is shown that the ultimate design of the allowable gross thermal loading density, and, thus, the layout of the underground workings is highly dependent upon the rock mass properties supplied as base line input data to the numerical models. Of the many input properties required, the thermal conductivity, the thermal expansion coefficient, and elastic moduli of the rock mass have, perhaps, the greatest effect on the calculation of induced temperatures, stresses, and displacements and, thus, repository design. To ensure that the design continues with confidence, field (in situ) values of input data must be obtained. The role of the Near-Surface Test Facility in situ testing in obtaining these basic required data is discussed

Quantitative comparison of the in situ microbial communities in different biomes

Energy Technology Data Exchange (ETDEWEB)

White, D.C. [Tennessee Univ., Knoxville, TN (United States)]|[Oak Ridge National Lab., TN (United States); Ringelberg, D.B.; Palmer, R.J. [Tennessee Univ., Knoxville, TN (United States). Center for Environmental Biotechnology

1995-12-31

A system to define microbial communities in different biomes requires the application of non-traditional methodology. Classical microbiological methods have severe limitations for the analysis of environmental samples. Pure-culture isolation, biochemical testing, and/or enumeration by direct microscopic counting are not well suited for the estimation of total biomass or the assessment of community composition within environmental samples. Such methods provide little insight into the in situ phenotypic activity of the extant microbiota since these techniques are dependent on microbial growth and thus select against many environmental microorganisms which are non- culturable under a wide range of conditions. It has been repeatedly documented in the literature that viable counts or direct counts of bacteria attached to sediment grains are difficult to quantitative and may grossly underestimate the extent of the existing community. The traditional tests provide little indication of the in situ nutritional status or for evidence of toxicity within the microbial community. A more recent development (MIDI Microbial Identification System), measure free and ester-linked fatty acids from isolated microorganisms. Bacterial isolates are identified by comparing their fatty acid profiles to the MIKI database which contains over 8000 entries. The application of the MIKI system to the analysis of environmental samples however, has significant drawbacks. The MIDI system was developed to identify clinical microorganisms and requires their isolation and culture on trypticase soy agar at 27{degrees}C. Since many isolates are unable to grow at these restrictive growth conditions, the system does not lend itself to identification of some environmental organisms. A more applicable methodology for environmental microbial analysis is based on the liquid extrication and separation of microbial lipids from environmental samples, followed by quantitative analysis using gas chromatography/

A FISH-based method for assessment of HER-2 amplification status in breast cancer circulating tumor cells following CellSearch isolation

Directory of Open Access Journals (Sweden)

Frithiof H

2016-11-01

Full Text Available Henrik Frithiof,1 Kristina Aaltonen,1 Lisa Rydén2,3 1Division of Oncology and Pathology, 2Division of Surgery, Department of Clinical Sciences Lund, Lund University, Lund, 3Department of Surgery, Skåne University Hospital, Malmö, Sweden Introduction: Amplification of the HER-2/neu (HER-2 proto-oncogene occurs in 10%–15% of primary breast cancer, leading to an activated HER-2 receptor, augmenting growth of cancer cells. Tumor classification is determined in primary tumor tissue and metastatic biopsies. However, malignant cells tend to alter their phenotype during disease progression. Circulating tumor cell (CTC analysis may serve as an alternative to repeated biopsies. The Food and Drug Administration-approved CellSearch system allows determination of the HER-2 protein, but not of the HER-2 gene. The aim of this study was to optimize a fluorescence in situ hybridization (FISH-based method to quantitatively determine HER-2 amplification in breast cancer CTCs following CellSearch-based isolation and verify the method in patient samples. Methods: Using healthy donor blood spiked with human epidermal growth factor receptor 2 (HER-2-positive breast cancer cell lines, SKBr-3 and BT-474, and a corresponding negative control (the HER-2-negative MCF-7 cell line, an in vitro CTC model system was designed. Following isolation in the CellSearch system, CTC samples were further enriched and fixed on microscope slides. Immunocytochemical staining with cytokeratin and 4',6-diamidino-2'-phenylindole dihydrochloride identified CTCs under a fluorescence microscope. A FISH-based procedure was optimized by applying the HER2 IQFISH pharmDx assay for assessment of HER-2 amplification status in breast cancer CTCs. Results: A method for defining the presence of HER-2 amplification in single breast cancer CTCs after CellSearch isolation was established using cell lines as positive and negative controls. The method was validated in blood from breast cancer patients

Comparison of in situ DGT measurement with ex situ methods for predicting cadmium bioavailability in soils with combined pollution to biotas.

Science.gov (United States)

Wang, Peifang; Liu, Cui; Yao, Yu; Wang, Chao; Wang, Teng; Yuan, Ye; Hou, Jun

2017-05-01

To assess the capabilities of the different techniques in predicting Cadmium (Cd) bioavailability in Cd-contaminated soils with the addition of Zn, one in situ technique (diffusive gradients in thin films; DGT) was compared with soil solution concentration and four widely used single-step extraction methods (acetic acid, EDTA, sodium acetate and CaCl 2 ). Wheat and maize were selected as tested species. The results demonstrated that single Cd-polluted soils inhibited the growth of wheat and maize significantly compared with control plants; the shoot and root biomasses of the plants both dropped significantly (P 0.9) between Cd concentrations in two plants and Cd bioavailability indicated by each method in soils. Consequently, the results indicated that the DGT technique could be regarded as a good predictor of Cd bioavailability to plants, comparable to soil solution concentration and the four single-step extraction methods. Because the DGT technique can offer in situ data, it is expected to be widely used in more areas.

Evaluation of isolation methods for pathogenic Yersinia enterocolitica from pig intestinal content.

Science.gov (United States)

Laukkanen, R; Hakkinen, M; Lundén, J; Fredriksson-Ahomaa, M; Johansson, T; Korkeala, H

2010-03-01

The aim of this study was to evaluate the efficiency of four isolation methods for the detection of pathogenic Yersinia enterocolitica from pig intestinal content. The four methods comprised of 15 isolation steps using selective enrichments (irgasan-ticarcillin-potassium chlorate and modified Rappaport broth) and mildly selective enrichments at 4 or 25 degrees C. Salmonella-Shigella-desoxycholate-calcium chloride agar, cefsulodin-irgasan-novobiocin agar were used as plating media. The most sensitive method detected 78% (53/68) of the positive samples. Individual isolation steps using cold enrichment as the only enrichment or as a pre-enrichment step with further selective enrichment showed the highest sensitivities (55-66%). All isolation methods resulted in high numbers of suspected colonies not confirmed as pathogenic Y. enterocolitica. Cold enrichment should be used in the detection of pathogenic Y. enterocolitica from pig intestinal contents. In addition, more than one parallel isolation step is needed. The study shows that depending on the isolation method used for Y. enterocolitica, the detected prevalence of Y. enterocolitica in pig intestinal contents varies greatly. More selective and sensitive isolation methods need to be developed for pathogenic Y. enterocolitica.

In-situ gamma spectrometry method for determination of environmental gamma dose

International Nuclear Information System (INIS)

Conti, Claudio de Carvalho

1995-07-01

This work tries to establish a methodology for germanium detectors calibration, normally used for in situ gamma ray spectrometry, for determining the environmental exposure rate in function of the energy of the incident photons. For this purpose a computer code has been developed, based on the stripping method, for the computational spectra analysis to calculate the contribution of the partial absorption of the gamma rays (Compton effect) in the active and nonactive parts of the detector. The resulting total absorption spectrum is then converted to fluence distribution in function of the energy for the photons reaching the detector, which is then used to calculate the exposure rate or kerma in air. The unfolding and fluency convention parameters are determined by detector calibration using point gamma sources. The method is validated by comparison of the results against the calculated exposure rate at a point of interest for the standards. This method is used for the direct measurement of the exposure rate distribution in function of the energy at the site, in situ measurement technic, leading to rapid results during an emergency situation and also used for indoor measurements. (author)

Laser Based In Situ Techniques: Novel Methods for Generating Extreme Conditions in TEM Samples

Energy Technology Data Exchange (ETDEWEB)

Taheri, M; Lagrange, T; Reed, B; Armstrong, M; Campbell, G; DeHope, W; Kim, J; King, W; Masiel, D; Browning, N

2008-02-25

The Dynamic Transmission Electron Microscope (DTEM) is introduced as a novel tool for in situ processing of materials. Examples of various types of dynamic studies outline the advantages and differences of laser-based heating in the DTEM in comparison to conventional (resistive) heating in situ TEM methods. We demonstrate various unique capabilities of the drive laser, namely, in situ processing of nanoscale materials, rapid and high temperature phase transformations, and controlled thermal activation of materials. These experiments would otherwise be impossible without the use of the DTEM drive laser. Thus, the potential of the DTEM to as a new technique to process and characterize the growth of a myriad of micro and nanostructures is demonstrated.

In-Situ Calibration of the CMS Electromagnetic Calorimeter

CERN Document Server

Futyan, D I

2003-01-01

The in-situ intercalibration of the lead tungstate crystals of the CMS electromagnetic calorimeter will be performed using 3 techniques: An energy flow method will be used at startup to intercalibrate to a precision of around 2% within about 3 hours. The energy/momentum measurement of isolated electrons from W decay will then be used to obtain the design goal precision of 0.5% within about 2 months. Global intercalibration of different regions of the calorimeter and the determination of the absolute energy scale will be performed using energetic electrons from Z->ee events.

Preservation methods for isolates of ascochyta blight fungi

Directory of Open Access Journals (Sweden)

Joanna Marcinkowska

2014-08-01

Full Text Available Isolates of ascochyta blight fungi, two of Ascochyta pisi, four of Mycosphaerella pinodes and four of Phoma pinodella were stored: A - on slants under mineral oil, B - on CN's medium agar disks, and as conidial suspension: C - in glycerine, D · in water. Viability and pathogenicity of recovered cultures after each consecutive year were assesed from 1991 to 1999. The compared parameters were first of all strongly influenced by the preservation method, but fungus species and number of years had a minor importance. The best for longer storage was method "A" because after 9 years the isolates were viable, highly pathogenic, and cultures recovered from them were clean. Thc method "C'' is good for short keeping (2-3 years, as conidia in vials need only small space and gave clean cultures.

Semiconductor/metal nanocomposites formed by in situ reduction method in multilayer thin films

International Nuclear Information System (INIS)

Song Yanli; Wang Enbo; Tian Chungui; Mao Baodong; Wang Chunlei

2009-01-01

A layer-by-layer adsorption and in situ reduction method was adopted for synthesizing semiconductor/metal nanocomposites in multilayer ultra-thin films. Alternate adsorption of ZnO nanoparticles modified with poly(ethyleneimine), hydrogentetrachloroaurate and poly(styrenesulfonate) sodium results in the formation of ZnO/AuCl 4 - -loaded multilayer films. In situ reduction of the incorporated metal ions by heating yields ZnO/Au nanocomposites in the films. UV-vis absorption spectroscopy and X-ray photoelectron spectroscopy were used to characterize the components of the composite films. UV-vis spectra indicate regular growth of the films. The electrochemistry behavior of the multilayer films was studied in detail on indium tin oxide electrode. The combined results suggest that the layer-by-layer adsorption and subsequent reduction method used here provides an effective way to synthesize ZnO/Au nanocomposites in the polymer matrix

[New isolation methods and phylogenetic diversity of actinobacteria from hypersaline beach in Aksu].

Science.gov (United States)

Zhang, Yao; Xia, Zhanfeng; Cao, Xinbo; Li, Jun; Zhang, Lili

2013-08-04

We explored 4 new methods to improve the isolation of actinobacterial resources from high salt areas. Optimized media based on 4 new strategies were used for isolating actinobacteria from hypersaline beaches. Glycerin-arginine, trehalose-creatine, glycerol-asparticacid, mannitol-casein, casein-mannitol, mannitol-alanine, chitosan-asparagineand GAUZE' No. 1 were used as basic media. New isolation strategy includes 4 methods: ten-fold dilution culture, simulation of the original environment, actinobacterial culture guided by uncultured molecular technology detected, and reference of actinobacterial media for brackish marine environment. The 16S rRNA genes of the isolates were amplified with bacterial universal primers. The results of 16S rRNA gene sequences were compared with sequences obtained from GenBank databases. We constructed phylogenetic tree with the neighbor-joining method. No actinobacterial strains were isolated by 8 media of control group, while 403 strains were isolated by new strategies. The isolates by new methods were members of 14 genera (Streptomyces, Streptomonospora, Saccharomonospora, Plantactinospora, Nocardia, Amycolatopsis, Glycomyces, Micromonospora, Nocardiopsis, Isoptericola, Nonomuraea, Thermobifida, Actinopolyspora, Actinomadura) of 10 families in 8 suborders. The most abundant and diverse isolates were the two suborders of Streptomycineae (69.96%) and Streptosporangineaesuborder (9.68%) within the phylum Actinobacteria, including 9 potential novel species. New isolation methods significantly improved the actinobacterial culturability of hypersaline areas, and obtained many potential novel species, which provided a new and more effective way to isolate actinobacteria resources in hypersaline environments.

Aerobic De-Epoxydation of Trichothecene Mycotoxins by a Soil Bacterial Consortium Isolated Using In Situ Soil Enrichment

Directory of Open Access Journals (Sweden)

Wei-Jie He

2016-09-01

Full Text Available Globally, the trichothecene mycotoxins deoxynivalenol (DON and nivalenol (NIV are among the most widely distributed mycotoxins that contaminate small grain cereals. In this study, a bacterial consortium, PGC-3, with de-epoxydation activity was isolated from soil by an in situ soil enrichment method. Screening of 14 soil samples that were sprayed with DON revealed that 4 samples were able to biotransform DON into de-epoxydized DON (dE-DON. Among these, the PGC-3 consortium showed the highest and most stable activity to biotransform DON into dE-DON and NIV into dE-NIV. PGC-3 exhibited de-epoxydation activity at a wide range of pH (5–10 and temperatures (20–37 °C values under aerobic conditions. Sequential subculturing with a continued exposure to DON substantially reduced the microbial population diversity of this consortium. Analyses of the 16S rDNA sequences indicated that PGC-3 comprised 10 bacterial genera. Among these, one species, Desulfitobacterium, showed a steady increase in relative abundance, from 0.03% to 1.55% (a 52-fold increase, as higher concentrations of DON were used in the subculture media, from 0 to 500 μg/mL. This study establishes the foundation to further develop bioactive agents that can detoxify trichothecene mycotoxins in cereals and enables for the characterization of detoxifying genes and their regulation.

Method for in situ determination cation exchange capacities of subsurface formations

International Nuclear Information System (INIS)

Fertl, W.H.; Welker, D.W.

1980-01-01

A method is disclosed for the in situ examination of each subsurface formation penetrated by a borehole to ascertain the cation exchange capacity of such formations within a geological region. Natural γ ray logging is used to develop signals functionally related to the total γ radiation and to the potassium-40, uranium and thorium energy-band radiations. A first borehole is traversed by a potential γ ray spectrometer to provide selected measurements of natural γ radiation. Core samples are taken from the logged formation and laboratory tests performed to determine the cation exchange capacity thereof. The cation exchange capacities thus are developed then correlated with selected parameters provided by the γ ray spectrometer to establish functional relationships. Cation exchange capacities of formations in subsequent boreholes within the region are then determined in situ by use of the natural γ ray spectrometer and these established relationships. (author)

An in-situ measuring method for planar straightness error

Science.gov (United States)

Chen, Xi; Fu, Luhua; Yang, Tongyu; Sun, Changku; Wang, Zhong; Zhao, Yan; Liu, Changjie

2018-01-01

According to some current problems in the course of measuring the plane shape error of workpiece, an in-situ measuring method based on laser triangulation is presented in this paper. The method avoids the inefficiency of traditional methods like knife straightedge as well as the time and cost requirements of coordinate measuring machine(CMM). A laser-based measuring head is designed and installed on the spindle of a numerical control(NC) machine. The measuring head moves in the path planning to measure measuring points. The spatial coordinates of the measuring points are obtained by the combination of the laser triangulation displacement sensor and the coordinate system of the NC machine, which could make the indicators of measurement come true. The method to evaluate planar straightness error adopts particle swarm optimization(PSO). To verify the feasibility and accuracy of the measuring method, simulation experiments were implemented with a CMM. Comparing the measurement results of measuring head with the corresponding measured values obtained by composite measuring machine, it is verified that the method can realize high-precise and automatic measurement of the planar straightness error of the workpiece.

Efficiency of consortium for in-situ bioremediation and CO2 evolution method of refines petroleum oil in microcosms study

OpenAIRE

Dutta, Shreyasri; Singh, Padma

2017-01-01

An in-situ bioremediation study was conducted in a laboratory by using mixed microbial consortium. An indigenous microbial consortium was developed by assemble of two Pseudomonas spp. and two Aspergillus spp. which were isolated from various oil contaminated sites of India. The laboratory feasibility study was conducted in a 225 m2 block. Six treatment options-Oil alone, Oil+Best remediater, Oil+Bacterial consortium, Oil+Fungal consortium, Oil+Mixed microbial consortium, Oil+Indigenous microf...

In-Situ Synchrotron X-ray Study of the Phase and Texture Evolution of Ceria and Superconductor Films Deposited by Chemical Solution Method

DEFF Research Database (Denmark)

Yue, Zhao; Grivel, Jean-Claude; He, Dong

2012-01-01

In situ synchrotron x-ray diffraction is used to study the phase and texture formation of ceria based films and superconductor films deposited by the chemical solution method on technical substrates. Combined analysis using in situ synchrotron x-ray diffraction, thermogravimetry/differential ther......In situ synchrotron x-ray diffraction is used to study the phase and texture formation of ceria based films and superconductor films deposited by the chemical solution method on technical substrates. Combined analysis using in situ synchrotron x-ray diffraction, thermogravimetry...

A fast method for large-scale isolation of phages from hospital ...

African Journals Online (AJOL)

This plaque-forming method could be adopted to isolate E. coli phage easily, rapidly and in large quantities. Among the 18 isolated E. coli phages, 10 of them had a broad host range in E. coli and warrant further study. Key words: Escherichia coli phages, large-scale isolation, drug resistance, biological properties.

Resource Isolation Method for Program’S Performance on CMP

Science.gov (United States)

Guan, Ti; Liu, Chunxiu; Xu, Zheng; Li, Huicong; Ma, Qiang

2017-10-01

Data center and cloud computing are more popular, which make more benefits for customers and the providers. However, in data center or clusters, commonly there is more than one program running on one server, but programs may interference with each other. The interference may take a little effect, however, the interference may cause serious drop down of performance. In order to avoid the performance interference problem, the mechanism of isolate resource for different programs is a better choice. In this paper we propose a light cost resource isolation method to improve program’s performance. This method uses Cgroups to set the dedicated CPU and memory resource for a program, aiming at to guarantee the program’s performance. There are three engines to realize this method: Program Monitor Engine top program’s resource usage of CPU and memory and transfer the information to Resource Assignment Engine; Resource Assignment Engine calculates the size of CPU and memory resource should be applied for the program; Cgroups Control Engine divide resource by Linux tool Cgroups, and drag program in control group for execution. The experiment result show that making use of the resource isolation method proposed by our paper, program’s performance can be improved.

Novel in-situ lamella fabrication technique for in-situ TEM.

Science.gov (United States)

Canavan, Megan; Daly, Dermot; Rummel, Andreas; McCarthy, Eoin K; McAuley, Cathal; Nicolosi, Valeria

2018-03-29

In-situ transmission electron microscopy is rapidly emerging as the premier technique for characterising materials in a dynamic state on the atomic scale. The most important aspect of in-situ studies is specimen preparation. Specimens must be electron transparent and representative of the material in its operational state, amongst others. Here, a novel fabrication technique for the facile preparation of lamellae for in-situ transmission electron microscopy experimentation using focused ion beam milling is developed. This method involves the use of rotating microgrippers during the lift-out procedure, as opposed to the traditional micromanipulator needle and platinum weld. Using rotating grippers, and a unique adhesive substance, lamellae are mounted onto a MEMS device for in-situ TEM annealing experiments. We demonstrate how this technique can be used to avoid platinum deposition as well as minimising damage to the MEMS device during the thinning process. Our technique is both a cost effective and readily implementable alternative to the current generation of preparation methods for in-situ liquid, electrical, mechanical and thermal experimentation within the TEM as well as traditional cross-sectional lamella preparation. Copyright © 2018 Elsevier B.V. All rights reserved.

Planning, developing, and fielding of thermal/structural interactions in situ tests for the Waste Isolation Pilot Plant (WIPP)

International Nuclear Information System (INIS)

Munson, D.E.; Matalucci, R.V.

1986-01-01

Large-scale, well-instrumented underground tests to determine in situ thermal/structural response of bedded salt are being constructed in the WIPP facility in southeastern New Mexico. These tests are an essential component of a broad research and development program to resolve thermal/structural issues, to validate long-term prediction methods, and to develop a design basis for a future repository. They are the result of an extensive planning and evaluation procedure to determine the appropriate test configuration. All details of the tests, including background, decisions, design, site operations, and testing organization are explained. These procedures may be useful in developing other in situ tests

Various methods for isolation of multipotent human periodontal ligament cells for regenerative medicine.

Science.gov (United States)

Tran, Ha Le Bao; Doan, Vu Nguyen; Le, Huong Thi Ngoc; Ngo, Lan Thi Quynh

2014-08-01

Periodontal ligament (PDL) is a specialized connective tissue that connects cementum and alveolar bone to maintain and support the teeth in situ and preserve tissue homeostasis. Recent studies have revealed the existence of stem cells in human dental tissues including periodontal ligament that play an important role, not only in the maintenance of the periodontium but also in promoting periodontal regeneration. In this study, human periodontal ligament cells (hPDLCs) were isolated by outgrowth and enzymatic dissociation methods. Expression of surface markers on PDLCs as human mesenchymal stem cells (MSCs) was identified by flow cytometry. In addition, proliferation and differentiation capacity of cultured cells to osteoblasts, adipocytes were evaluated. As a result, we successfully cultured cells from the human periodontal ligament tissues. PDLCs express mesenchymal stem cell (MSC) markers such as CD44, CD73, and CD90 and do not express CD34, CD45, and HLA-DR. PDLCs also possess the multipotential to differentiate into various types of cells, such as osteoblast and adipocytes, in vitro. Therefore, these cells have high potential to serve as materials for tissue engineering, especially dental tissue engineering.

Isolation of cancer cells by "in situ" microfluidic biofunctionalization protocols

DEFF Research Database (Denmark)

De Vitis, Stefania; Matarise, Giuseppina; Pardeo, Francesca

2014-01-01

The aim of this work is the development of a microfluidic immunosensor for the immobilization of cancer cells and their separation from healthy cells by using "in situ" microfluidic biofunctionalization protocols. These protocols allow to link antibodies on microfluidic device surfaces and can be...

Water feeding method upon reactor isolation

International Nuclear Information System (INIS)

Sasaki, Koichi; Takahara, Kuniaki; Hamamura, Kenji; Arakawa, Masahiro.

1990-01-01

The present invention concerns a method of feeding water upon reactor isolation in a plural loop type reactor having a plurality of reactor cooling systems. Water can be injected to a plurality of pools even if the pressure between the pools is not balanced and the water level in the reactor cooling system is optimally controlled. That is, water can be injected in accordance with the amount required for each of the pools by setting the opening of a turbine inlet steam control valve to somewhat higher than the cooling system pressure of the highest pressure loop. Water feeding devices upon reactor isolation were required by the same number as that for the reactor cooling systems. Whereas since pumps and turbines are used in common without worsening the water injection controllability to each of the loops according to the method of this invention and, accordingly, the cost performance can be improved. Further, since the opening degree of the turbine inlet steam control valve is controlled while making the difference pressure constant between the turbine inlet pressure and the pump exhaust pressure, the amount of the turbine exhausted steams can be reduced and, further, water injection controllability of the flow rate control valve in the injection line is improved. (I.S.)

Development of an evaluation method for seismic isolation systems of nuclear power facilities. Development of crossover piping design method for seismic isolation systems

International Nuclear Information System (INIS)

Otoyo, Teruyoshi; Otani, Akihito; Otani, Akihito; Fukushima, Shunsuke; Jimbo, Masakazu; Yamamoto, Tomofumi; Sakakida, Takaaki; Onishi, Shigenobu

2014-01-01

In the conceptual design of seismic isolation systems of nuclear power facilities, there exist two types of installation. The first type is to isolate both the reactor and the turbine buildings, the other is to isolate only the reactor building. In the latter type, the crossover piping, which installed between the isolated and the non-isolated buildings, is excited and deformed by the different motions of those buildings. In this study, shaking tests of 1/10 scaled model of the main steam piping and FEM analyses under multiple support excitation conditions have been performed to investigate the vibration behavior of the crossover piping. It was confirmed that modal time-history analyses could be in good agreement with the shaking test results. Also, Numerous combination methods were investigated by comparing response spectrum analyses and modal time-history analyses. In conclusion, response spectrum analyses using SRSS combinations could correspond to time-history analyses. (author)

Evaluation of Methods for In-Situ Calibration of Field-Deployable Microphone Phased Arrays

Science.gov (United States)

Humphreys, William M.; Lockard, David P.; Khorrami, Mehdi R.; Culliton, William G.; McSwain, Robert G.

2017-01-01

Current field-deployable microphone phased arrays for aeroacoustic flight testing require the placement of hundreds of individual sensors over a large area. Depending on the duration of the test campaign, the microphones may be required to stay deployed at the testing site for weeks or even months. This presents a challenge in regards to tracking the response (i.e., sensitivity) of the individual sensors as a function of time in order to evaluate the health of the array. To address this challenge, two different methods for in-situ tracking of microphone responses are described. The first relies on the use of an aerial sound source attached as a payload on a hovering small Unmanned Aerial System (sUAS) vehicle. The second relies on the use of individually excited ground-based sound sources strategically placed throughout the array pattern. Testing of the two methods was performed in microphone array deployments conducted at Fort A.P. Hill in 2015 and at Edwards Air Force Base in 2016. The results indicate that the drift in individual sensor responses can be tracked reasonably well using both methods. Thus, in-situ response tracking methods are useful as a diagnostic tool for monitoring the health of a phased array during long duration deployments.

Comparison of three 15N methods to correct for microbial contamination when assessing in situ protein degradability of fresh forages.

Science.gov (United States)

Kamoun, M; Ammar, H; Théwis, A; Beckers, Y; France, J; López, S

2014-11-01

The use of stable (15)N as a marker to determine microbial contamination in nylon bag incubation residues to estimate protein degradability was investigated. Three methods using (15)N were compared: (15)N-labeled forage (dilution method, LF), (15)N enrichment of rumen solids-associated bacteria (SAB), and (15)N enrichment of rumen liquid-associated bacteria (LAB). Herbage from forages differing in protein and fiber contents (early-cut Italian ryegrass, late-cut Italian ryegrass, and red clover) were freeze-dried and ground and then incubated in situ in the rumen of 3 steers for 3, 6, 12, 24, and 48 h using the nylon bag technique. The (15)N-labeled forages were obtained by fertilizing the plots where herbage was grown with (15)NH4 (15)NO3. Unlabeled forages (obtained from plots fertilized with NH4NO3) were incubated at the same time that ((15)NH4)2SO4 was continuously infused into the rumen of the steers, and then pellets of labeled SAB and LAB were isolated by differential centrifugation of samples of ruminal contents. The proportion of bacterial N in the incubation residues increased from 0.09 and 0.45 g bacterial N/g total N at 3 h of incubation to 0.37 and 0.85 g bacterial N/g total N at 48 h of incubation for early-cut and late-cut ryegrass, respectively. There were differences (P forage (late-cut ryegrass) was 0.51, whereas the corrected values were 0.85, 0.84, and 0.77 for the LF, SAB, and LAB methods, respectively. With early-cut ryegrass and red clover, the differences between uncorrected and corrected values ranged between 6% and 13%, with small differences among the labeling methods. Generally, methods using labeled forage or labeled SAB and LAB provided similar corrected degradability values. The accuracy in estimating the extent of degradation of protein in the rumen from in situ disappearance curves is improved when values are corrected for microbial contamination of the bag residue.

In situ generation of diazonium cations in organic electrolyte for electrochemical modification of electrode surface

International Nuclear Information System (INIS)

Baranton, Steve; Belanger, Daniel

2008-01-01

The modification of glassy carbon electrode was achieved by electrochemical reduction of in situ generated diazonium cations in acetonitrile. The in situ generation of 4-nitrophenyl diazonium cations in acetonitrile was investigated by spectroscopic methods. UV-visible spectroscopy revealed slow kinetics for the reaction of 4-nitroaniline with tert-butylnitrite in acetonitrile to form the corresponding diazonium cation. As a result, a coupling reaction, which implies a consumption of the amine and loss of the already formed diazonium cations, was evidenced by 1 H NMR spectroscopy. This spectroscopic study allowed the optimization of the in situ diazonium cations generation prior to the modification step. The electrochemical modification of the carbon electrodes with 4-nitrophenyl, 4-bromophenyl and anthraquinone groups was characterized by cyclic voltammetry and the resulting grafted layer were characterized by electrochemical techniques. The cyclic voltammetric behaviour during the electrochemical grafting was very similar to the one observed for an isolated diazonium salt dissolved in acetonitrile. In the case of the anthraquinone-modified electrode, the use of acetonitrile, into which the corresponding amine is soluble but not in aqueous media, allowed for its grafting by the in situ approach. The barrier properties of these grafted layers are similar to those obtained from isolated diazonium salts. Finally, the chemical composition of the grafted layers was determined by X-ray photoelectron spectroscopy and surface coverage in the range 5-7 x 10 -10 mol cm -2 was estimated for films grown in our experimental conditions

In situ generation of diazonium cations in organic electrolyte for electrochemical modification of electrode surface

Energy Technology Data Exchange (ETDEWEB)

Baranton, Steve [Departement de Chimie, Universite du Quebec a Montreal, Case Postale 8888, succursale Centre-Ville, Montreal (Quebec), H3C 3P8 (Canada); Belanger, Daniel [Departement de Chimie, Universite du Quebec a Montreal, Case Postale 8888, succursale Centre-Ville, Montreal (Quebec), H3C 3P8 (Canada)], E-mail: belanger.daniel@uqam.ca

2008-10-01

The modification of glassy carbon electrode was achieved by electrochemical reduction of in situ generated diazonium cations in acetonitrile. The in situ generation of 4-nitrophenyl diazonium cations in acetonitrile was investigated by spectroscopic methods. UV-visible spectroscopy revealed slow kinetics for the reaction of 4-nitroaniline with tert-butylnitrite in acetonitrile to form the corresponding diazonium cation. As a result, a coupling reaction, which implies a consumption of the amine and loss of the already formed diazonium cations, was evidenced by {sup 1}H NMR spectroscopy. This spectroscopic study allowed the optimization of the in situ diazonium cations generation prior to the modification step. The electrochemical modification of the carbon electrodes with 4-nitrophenyl, 4-bromophenyl and anthraquinone groups was characterized by cyclic voltammetry and the resulting grafted layer were characterized by electrochemical techniques. The cyclic voltammetric behaviour during the electrochemical grafting was very similar to the one observed for an isolated diazonium salt dissolved in acetonitrile. In the case of the anthraquinone-modified electrode, the use of acetonitrile, into which the corresponding amine is soluble but not in aqueous media, allowed for its grafting by the in situ approach. The barrier properties of these grafted layers are similar to those obtained from isolated diazonium salts. Finally, the chemical composition of the grafted layers was determined by X-ray photoelectron spectroscopy and surface coverage in the range 5-7 x 10{sup -10} mol cm{sup -2} was estimated for films grown in our experimental conditions.

Improvements in in-situ filter test methods using a total light-scattering detector

International Nuclear Information System (INIS)

Marshall, M.; Stevens, D.C.

1986-01-01

This paper presents research aimed at providing useful data on a commonly used technique; a DOP (di-2-ethylhexyl phthalate) aerosol and a total light-scattering photometer. Methods of increasing the sensitivity of this technique are described. Alternative methods of in-situ filter testing are also considered. The sensitivity of a typical, modern, total light-scattering photometer, as a function of particle diameter, has a broad maximum in mass terms between 0.1 and 0.4 um. At its maximum usable sensitivity the instrument can detect approx. 1 particle/cm 3 . This response can be explained by light scattering theory and particle loss in the instrument inlet. The mass median diameter of the aerosols produced by various DOP generators varies from 0.2 to 1.0μm. Experiments with good quality HEPA filters indicate a maximum penetration for particles of 0.15 - 0.2μm. Details of the studies are given and the consequences discussed. It is shown that filter penetration of -3 % can be measured in-situ with existing equipment. Methods of extending the sensitivity to measure a penetration of approx.10 -5 % are described. (author)

Synchronous cecal adenocarcinoma and multiple colonic in situ carcinomas in hamartomatous polyps in a case of isolated Peutz–Jeghers syndrome

Directory of Open Access Journals (Sweden)

Yahia Z Gad

2011-03-01

Full Text Available Yahia Z Gad1, Doaa H Bakr1, Mohammad G El-Ebeidy21Department of Internal Medicine, 2Department of Surgery, Mansoura Specialized Medical Hospital, Mansoura University, Mansoura, EgyptBackground: Peutz–Jeghers syndrome (PJS is a rare autosomal dominant disease characterized by mucocutaneous pigmentation and hamartomatous polyps of the entire gastrointestinal tract. A Peutz–Jeghers polyp (PJP in a patient without pigmentation or a family history of the disease is called an isolated or solitary PJP. Individuals with PJS carry a very high risk of developing gastrointestinal (GI as well as extra-GI malignancies. This case report documents lesion multiplicity and their malignant potential in a young patient with PJS presenting in a serious condition for the first time.Case report: An 18-year-old female Egyptian patient was admitted with hematochezia and remarkable anemia. After appropriate resuscitation and consent, colonoscopic evaluation revealed seven pedunculated colonic polyps at the ascending and the transverse colon, and numerous variable-sized sessile polyps were scattered all over the colon. To establish hemostasis, endoscopic polypectomy for pedunculated polyps and argon plasma photocoagulation for the bleeding sessile polyps were performed. Histopathological examination revealed cecal adenocarcinoma in one specimen and two simultaneous in situ carcinoma at the transverse and the sigmoid colon in the mucosae of the excised histologically proven hamartomatous polyps. Additionally, one focal in situ carcinoma in the resected colon was detected.Conclusions: When considering the family history, serious GI neoplastic lesions may be unmasked in young patients with PJS who present with hematochezia, even in the absence of its characteristic mucocutaneous pigmented lesions. GI endoscopic surveillance programs should be adopted for diagnosed cases of PJS and their families. Genetic prenatal screening for early detection is the best option for

Understanding the degree of condensation of phenolic and etherified C-9 units of in situ lignins.

Science.gov (United States)

Nanayakkara, Bernadette; Manley-Harris, Merilyn; Suckling, Ian D

2011-12-14

A novel approach for the quantification of the degree of condensation at the C(5) position of etherified and phenolic phenylpropane (C-9) units of in situ lignin is described. This is achieved by degrading unmethylated and methylated wood by thioacidolysis and analyzing the resultant product mixtures by quantitative (31)P NMR spectroscopy. Applying this new method to compression wood and normal wood from Pinus radiata showed that, whereas 41-47% of etherified guaiacyl C-9 units are condensed at the C(5) position, almost all phenolic guaiacyl C-9 units exist as uncondensed moieties. Analysis of milled wood lignin (MWL) isolated from the same wood by (31)P NMR spectroscopy before and after thioacidolysis showed that the phenolic guaiacyl C-9 units were more condensed than those in the in situ lignin. This is likely due to partial cleavage of the more condensed etherified linkages during the lignin isolation, leading to a relative increase in condensed phenolic guaiacyl C-9 units.

Antifungal susceptibility testing of vaginal candida isolates: the broth microdilution method

Directory of Open Access Journals (Sweden)

Mahmoudi Rad M

2010-02-01

Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Vulvovaginal candidiasis is a common mucosal infection among immunocompetent, healthy women, and is caused by opportunistic yeasts that belong to genus Candida. In this study, we isolated and identified the Candida species in the vagina of patients who admitted in Gynecology Department of Mahdieh Hospital in Tehran, Iran to evaluate the in vitro activities of fluconazole, miconazole, itraconazole and flucytosine against 191 clinical Candida isolates by the NCCLS microdilution method."n"nMethods: 191 Candida were isolated from vaginal secretions and identified with conventional mycological methods in the diagnosis of Candida species. The identity of all strains was confirmed genotypically by multiplex PCR. In vitro susceptibility testing of vaginal Candida isolates was performed by the NCCLS broth microdilution method. The results were read at 48 h."n"nResults: Most C. albicans isolates (>90% were sensitive in vitro to the antifungal agents tested. Most C. glabrata isolates showed sensitivity to miconazole and then flucytosine while they were more resistant to Itraconazole and fluconazole. Many isolates of C. tropicalis were susceptible to miconazole and then fluconazole. They showed a little resistance to

Carbapenem inactivation: a very affordable and highly specific method for phenotypic detection of carbapenemase-producing Pseudomonas aeruginosa isolates compared with other methods.

Science.gov (United States)

Akhi, Mohammad Taghi; Khalili, Younes; Ghotaslou, Reza; Kafil, Hossein Samadi; Yousefi, Saber; Nagili, Behroz; Goli, Hamid Reza

2017-06-01

This investigation was undertaken to compare phenotypic and molecular methods for detection of carbapenemase-producing Pseudomonas aeruginosa. A total of 245 non-duplicated isolates of P. aeruginosa were collected from hospitalized patients. Disc diffusion method was used to identify carbapenem-resistant bacteria. Three phenotypic methods, including Modified Hodge Test (MHT), Modified Carba NP (MCNP) test and Carbapenem Inactivation Method (CIM) were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was used to detect carbapenemase encoding genes. Of 245 P. aeruginosa isolates investigated, 121 isolates were carbapenem-resistant. Among carbapenem-resistant isolates, 40, 39 and 35 isolates exhibited positive results using MHT, MCNP test and CIM, respectively. PCR indicated the presence of carbapenemase genes in 35 of carbapenem-resistant isolates. MHT showed low sensitivity and specificity for carbapenemase detection among P. aeruginosa isolates in comparison to PCR. CIM was most affordable and highly specific than MCNP test compared with the molecular method.

IN-SITU MEASURING METHOD OF RADON AND THORON DIFFUSION COEFFICIENT IN SOIL

Directory of Open Access Journals (Sweden)

V.S. Yakovleva

2014-06-01

Full Text Available A simple and valid in-situ measurement method of effective diffusion coefficient of radon and thoron in soil and other porous materials was designed. The analysis of numerical investigation of radon and thoron transport in upper layers of soil revealed that thoron flux density from the earth surface does not depend on soil gas advective velocity and varies only with diffusion coefficient changes. This result showed the advantages of thoron using versus radon using in the suggested method. The comparison of the new method with existing ones previously developed. The method could be helpful for solving of problems of radon mass-transport in porous media and gaseous exchange between soil and atmosphere.

An optimized method for mouse liver sinusoidal endothelial cell isolation

Energy Technology Data Exchange (ETDEWEB)

Meyer, Jeremy, E-mail: jeremy.meyer@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Lacotte, Stéphanie, E-mail: stephanie.lacotte@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Morel, Philippe, E-mail: philippe.morel@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Gonelle-Gispert, Carmen, E-mail: carmen.gonelle@unige.ch [Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland); Bühler, Léo, E-mail: leo.buhler@hcuge.ch [Division of Digestive and Transplantation Surgery, University Hospitals of Geneva, Rue Gabrielle-Perret-Gentil 4, 1211 Genève 14 (Switzerland); Unit of Surgical Research, University of Geneva, Rue Michel-Servet 1, 1206 Genève (Switzerland)

2016-12-10

The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic �

An optimized method for mouse liver sinusoidal endothelial cell isolation

International Nuclear Information System (INIS)

Meyer, Jeremy; Lacotte, Stéphanie; Morel, Philippe; Gonelle-Gispert, Carmen; Bühler, Léo

2016-01-01

The objective of the present study was to develop an accurate and reproducible method for liver sinusoidal endothelial cell (LSEC) isolation in mice. Non-parenchymal cells were isolated using a modified two-step collagenase digestion combined with Optiprep density gradient centrifugation. LSEC were further purified using two prevalent methods, short-term selective adherence and CD146+ magnetic-activated cell sorting (MACS), and compared in terms of cell yield, viability and purity to our purification technique using CD11b cell depletion combined with long-term selective adherence. LSEC purification using our technique allowed to obtain 7.07±3.80 million LSEC per liver, while CD146+ MACS and short-term selective adherence yielded 2.94±1.28 and 0.99±0.66 million LSEC, respectively. Purity of the final cell preparation reached 95.10±2.58% when using our method. In contrast, CD146+ MACS and short-term selective adherence gave purities of 86.75±3.26% and 47.95±9.82%, respectively. Similarly, contamination by non-LSEC was the lowest when purification was performed using our technique, with a proportion of contaminating macrophages of only 1.87±0.77%. Further, isolated cells analysed by scanning electron microscopy presented typical LSEC fenestrations organized in sieve plates, demonstrating that the technique allowed to isolate bona fide LSEC. In conclusion, we described a reliable and reproducible technique for the isolation of high yields of pure LSEC in mice. This protocol provides an efficient method to prepare LSEC for studying their biological functions. - Highlights: • This protocol provides an efficient method to prepare primary mouse LSEC for studying their biological functions. • The liver cell dispersion step was improved by performing a retrograde cannulation of the liver. • The cell yield and the purity obtained were higher than comparative techniques in mice. • Contaminating macrophages were removed by introducing a CD11b- magnetic �

A practical method for in-situ thickness determination using energy distribution of beta particles

International Nuclear Information System (INIS)

Yalcin, S.; Gurler, O.; Gundogdu, O.; Bradley, D.A.

2012-01-01

This paper discusses a method to determine the thickness of an absorber using the energy distribution of beta particles. An empirical relationship was obtained between the absorber thickness and the energy distribution of beta particles transmitted through. The thickness of a polyethylene radioactive source cover was determined by exploiting this relationship, which has largely been left unexploited allowing us to determine the in-situ cover thickness of beta sources in a fast, cheap and non-destructive way. - Highlights: ► A practical and in-situ unknown cover thickness determination ► Cheap and readily available compared to other techniques. ► Beta energy spectrum.

A practical method for in-situ thickness determination using energy distribution of beta particles

Energy Technology Data Exchange (ETDEWEB)

Yalcin, S., E-mail: syalcin@kastamonu.edu.tr [Kastamonu University, Education Faculty, 37200 Kastamonu (Turkey); Gurler, O. [Physics Department, Faculty of Arts and Sciences, Uludag University, Gorukle Campus, 16059 Bursa (Turkey); Gundogdu, O. [Kocaeli University, Umuttepe Campus, 41380 Kocaeli (Turkey); Bradley, D.A. [CNRP, Department of Physics, Faculty of Engineering and Physical Sciences, University of Surrey, Guildford GU2 7XH (United Kingdom)

2012-01-15

This paper discusses a method to determine the thickness of an absorber using the energy distribution of beta particles. An empirical relationship was obtained between the absorber thickness and the energy distribution of beta particles transmitted through. The thickness of a polyethylene radioactive source cover was determined by exploiting this relationship, which has largely been left unexploited allowing us to determine the in-situ cover thickness of beta sources in a fast, cheap and non-destructive way. - Highlights: Black-Right-Pointing-Pointer A practical and in-situ unknown cover thickness determination Black-Right-Pointing-Pointer Cheap and readily available compared to other techniques. Black-Right-Pointing-Pointer Beta energy spectrum.

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

OpenAIRE

Pirttila A. M.; Kozyrovska N. O.; Ovcharenko L. P.; Podolich O. V.

2009-01-01

A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290) which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH). The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the pla...

In situ anisotropic parameter determination using refraction seismic and VSP methods

Energy Technology Data Exchange (ETDEWEB)

Leslie, J.M.; Lawton, D.C. (Calgary Univ., AB (Canada))

1999-01-01

A prime concern in the time-to-depth conversion of reflection seismic data is seismic anisotropy, because it can produce velocity anomalies in seismic data that mimic the structural plays of interest to the petroleum prospector in both size and shape. Ongoing techniques of time-to-depth conversion of P-wave seismic data do not handle the travel time and velocity distortions caused by seismic anisotropy, particularly in areas of complex geologic structures. To address this problem, the first step is to know which rock units are anisotropic and measure their anisotropic parameters. Laboratory means are available, but there are problems with these mainly with shales because of their fissile nature. In situ measurements are preferable because they yield a more robust value, and at the University of Calgary such measurements were undertaken using refraction seismic and vertical seismic profiling (VSP) methods. Results indicate that the two Thomsen anisotropic parameters of interest can be determined from the VSP experiment, but these values are slightly less than those obtained using the refraction technique. This may be because of the sensitivity of the shot statics which arises from the direct travel time measurement of the technique. The experiment yields another method to measure velocity anisotropy, in situ, where steeply dipping strata outcrop, which allows for the accurate measurement of the anisotropic parameters for use in depth migration routines. 4 refs.

In situ anisotropic parameter determination using refraction seismic and VSP methods

Energy Technology Data Exchange (ETDEWEB)

Leslie, J.M.; Lawton, D.C. [Calgary Univ., AB (Canada)

1999-11-01

A prime concern in the time-to-depth conversion of reflection seismic data is seismic anisotropy, because it can produce velocity anomalies in seismic data that mimic the structural plays of interest to the petroleum prospector in both size and shape. Ongoing techniques of time-to-depth conversion of P-wave seismic data do not handle the travel time and velocity distortions caused by seismic anisotropy, particularly in areas of complex geologic structures. To address this problem, the first step is to know which rock units are anisotropic and measure their anisotropic parameters. Laboratory means are available, but there are problems with these mainly with shales because of their fissile nature. In situ measurements are preferable because they yield a more robust value, and at the University of Calgary such measurements were undertaken using refraction seismic and vertical seismic profiling (VSP) methods. Results indicate that the two Thomsen anisotropic parameters of interest can be determined from the VSP experiment, but these values are slightly less than those obtained using the refraction technique. This may be because of the sensitivity of the shot statics which arises from the direct travel time measurement of the technique. The experiment yields another method to measure velocity anisotropy, in situ, where steeply dipping strata outcrop, which allows for the accurate measurement of the anisotropic parameters for use in depth migration routines. 4 refs.

Comparison of Two Methods of Ruminal Digestibility Determination (in situ and in vitro of Pomegranate Seed Pulp

Directory of Open Access Journals (Sweden)

Mohammad Hassan Fathi Nasri

2016-08-01

Full Text Available Introduction Using of agro-industrial by-products in diet of livestock not only reduces the production costs but also can dwindle the use of human foods in animal nutrition and competition between human and livestock. Thus, proper use of these by-products in animal nutrition and identification of new and low cost feed resources may be one of the priorities in livestock husbandry of our country. Pomegranate seed pulp (PSP is a by-product of the industrial decoction of pomegranate. Iran with annually production of more than 900 tons of pomegranates is one of the most important regions of pomegranate cultivation and PSP produced from pomegranate processing factories may be extensively used in animal nutrition. Digestibility determination of feeds is one of the most effective ways to evaluate their nutritional value. In addition, there is a strong relationship between feeds digestibility and performance of animal. There are in vivo, in vitro and in situ methods to determine the digestibility of feeds. Although in vivo methods are reference for digestibility values of feeds and are of high precision but they are usually expensive and time consuming. In addition, these methods do not provide any information related to ruminal degradability kinetic of nutrients. The aim of this study was comparison of two methods of ruminal degradability determination methods namely, in situ (nylon bag and in vitro (Daisy incubator, using dried and ensiled pomegranate seed pulp (PSP. Materials and Methods The PSP used in this study was prepared from Anaryan Co. in Ferdows, Iran. Decocted pomegranate was a mixture of Yazd varieties which were harvested at early autumn of 1389. Two types of PSP as dried and ensiled were used to compare the methods of ruminal digestibility determination, pervasively. The PSP silage was prepared by ensiling of wet PSP (containing 475 g/kg DM in 3 kg bins (4 replicatesand with density of 650 kg wet PSP per cubic meter for 60 days and dried

In-situ gamma spectrometry method for determination of environmental gamma dose; Metodo de espectrometria gamma in situ para determinacao de dose gama ambiental

Energy Technology Data Exchange (ETDEWEB)

Conti, Claudio de Carvalho

1995-07-15

This work tries to establish a methodology for germanium detectors calibration, normally used for in situ gamma ray spectrometry, for determining the environmental exposure rate in function of the energy of the incident photons. For this purpose a computer code has been developed, based on the stripping method, for the computational spectra analysis to calculate the contribution of the partial absorption of the gamma rays (Compton effect) in the active and nonactive parts of the detector. The resulting total absorption spectrum is then converted to fluence distribution in function of the energy for the photons reaching the detector, which is then used to calculate the exposure rate or kerma in air. The unfolding and fluency convention parameters are determined by detector calibration using point gamma sources. The method is validated by comparison of the results against the calculated exposure rate at a point of interest for the standards. This method is used for the direct measurement of the exposure rate distribution in function of the energy at the site, in situ measurement technic, leading to rapid results during an emergency situation and also used for indoor measurements. (author)

A method for isolating identifying and culturing of rat trachea-bronchia epithelial cells

International Nuclear Information System (INIS)

Cui Fengmei; Su Shibiao; Nie Jihua; Li Bingyan; Tong Jian

2005-01-01

Objective: To explore a method for isolating identifying and culturing the rat trachea-bronchia epithelial cells. Methods: The rat trachea-bronchia epithelial cells were isolated by digestion with pronase and brushing with cell brush, identified using confocul and cultured in entire F12 media with no serum. Results: With this method, cells in high purity and high viability could be obtained, and about 10 6 cells per rat. The cells grow well in entire F12 media with no serum. Conclusion: The method is useful for isolating rate trachea-bronchia epithelial cells and the entire F12 media with no serum is effective for culturing. (authors)

Intercomparison of analysis methods for seismically isolated nuclear structures. Part 1: Advanced test data and numerical methods. Working material

International Nuclear Information System (INIS)

1993-01-01

The purpose of the meeting was to review proposed contributions from CRP participating organizations to discuss in detail the experimental data on seismic isolators, to review the numerical methods for the analysis of the seismic isolators, and to perform a first comparison of the calculation results. The aim of the CRP was to validate the reliable numerical methods used for both detailed evaluation of dynamic behaviour of isolation devices and isolated nuclear structures of different nuclear power plant types. The full maturity of seismic isolation for nuclear applications was stressed, as well as the excellent behaviour of isolated structures during the recent earthquakes in Japan and the USA. Participants from Italy, USA, Japan, Russian federation, Republic of Korea, United Kingdom, India and European Commission have presented overview papers on the present programs and their status of contribution to the CRP

Intercomparison of analysis methods for seismically isolated nuclear structures. Part 1: Advanced test data and numerical methods. Working material

Energy Technology Data Exchange (ETDEWEB)

NONE

1993-07-01

The purpose of the meeting was to review proposed contributions from CRP participating organizations to discuss in detail the experimental data on seismic isolators, to review the numerical methods for the analysis of the seismic isolators, and to perform a first comparison of the calculation results. The aim of the CRP was to validate the reliable numerical methods used for both detailed evaluation of dynamic behaviour of isolation devices and isolated nuclear structures of different nuclear power plant types. The full maturity of seismic isolation for nuclear applications was stressed, as well as the excellent behaviour of isolated structures during the recent earthquakes in Japan and the USA. Participants from Italy, USA, Japan, Russian federation, Republic of Korea, United Kingdom, India and European Commission have presented overview papers on the present programs and their status of contribution to the CRP.

Novel RNA hybridization method for the in situ detection of ETV1, ETV4, and ETV5 gene fusions in prostate cancer.

Science.gov (United States)

Kunju, Lakshmi P; Carskadon, Shannon; Siddiqui, Javed; Tomlins, Scott A; Chinnaiyan, Arul M; Palanisamy, Nallasivam

2014-09-01

The genetic basis of 50% to 60% of prostate cancer (PCa) is attributable to rearrangements in E26 transformation-specific (ETS) (ERG, ETV1, ETV4, and ETV5), BRAF, and RAF1 genes and overexpression of SPINK1. The development and validation of reliable detection methods are warranted to classify various molecular subtypes of PCa for diagnostic and prognostic purposes. ETS gene rearrangements are typically detected by fluorescence in situ hybridization and reverse-transcription polymerase chain reaction methods. Recently, monoclonal antibodies against ERG have been developed that detect the truncated ERG protein in immunohistochemical assays where staining levels are strongly correlated with ERG rearrangement status by fluorescence in situ hybridization. However, specific antibodies for ETV1, ETV4, and ETV5 are unavailable, challenging their clinical use. We developed a novel RNA in situ hybridization-based assay for the in situ detection of ETV1, ETV4, and ETV5 in formalin-fixed paraffin-embedded tissues from prostate needle biopsies, prostatectomy, and metastatic PCa specimens using RNA probes. Further, with combined RNA in situ hybridization and immunohistochemistry we identified a rare subset of PCa with dual ETS gene rearrangements in collisions of independent tumor foci. The high specificity and sensitivity of RNA in situ hybridization provides an alternate method enabling bright-field in situ detection of ETS gene aberrations in routine clinically available PCa specimens.

Method for in situ or ex situ bioremediation of hexavalent chromium contaminated soils and/or groundwater

Science.gov (United States)

Turick, Charles E.; Apel, William W.

1997-10-28

A method of reducing the concentration of Cr(VI) in a liquid aqueous residue comprises the steps of providing anaerobic Cr(VI) reducing bacteria, mixing the liquid aqueous residue with a nutrient medium to form a mixture, and contacting the mixture with the anaerobic Cr(VI) reducing bacteria such that Cr(VI) is reduced to Cr(III). The anaerobic Cr(VI) reducing bacteria appear to be ubiquitous in soil and can be selected by collecting a soil sample, diluting the soil sample with a sterile diluent to form a diluted sample, mixing the diluted sample with an effective amount of a nutrient medium and an effective amount of Cr(VI) to form a mixture, and incubating the mixture in the substantial absence of oxygen such that growth of Cr(VI) sensitive microorganisms is inhibited and growth of the anaerobic Cr(VI) reducing bacteria is stimulated. A method of in situ bioremediation of Cr(VI) contaminated soil and/or groundwater is also disclosed.

A rapid and low-cost DNA extraction method for isolating ...

African Journals Online (AJOL)

The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA ...

Estimation of In Situ Stresses with Hydro-Fracturing Tests and a Statistical Method

Science.gov (United States)

Lee, Hikweon; Ong, See Hong

2018-03-01

At great depths, where borehole-based field stress measurements such as hydraulic fracturing are challenging due to difficult downhole conditions or prohibitive costs, in situ stresses can be indirectly estimated using wellbore failures such as borehole breakouts and/or drilling-induced tensile failures detected by an image log. As part of such efforts, a statistical method has been developed in which borehole breakouts detected on an image log are used for this purpose (Song et al. in Proceedings on the 7th international symposium on in situ rock stress, 2016; Song and Chang in J Geophys Res Solid Earth 122:4033-4052, 2017). The method employs a grid-searching algorithm in which the least and maximum horizontal principal stresses ( S h and S H) are varied, and the corresponding simulated depth-related breakout width distribution as a function of the breakout angle ( θ B = 90° - half of breakout width) is compared to that observed along the borehole to determine a set of S h and S H having the lowest misfit between them. An important advantage of the method is that S h and S H can be estimated simultaneously in vertical wells. To validate the statistical approach, the method is applied to a vertical hole where a set of field hydraulic fracturing tests have been carried out. The stress estimations using the proposed method were found to be in good agreement with the results interpreted from the hydraulic fracturing test measurements.

A whole biodiesel conversion process combining isolation, cultivation and in situ supercritical methanol transesterification of native microalgae.

Science.gov (United States)

Jazzar, Souhir; Quesada-Medina, Joaquín; Olivares-Carrillo, Pilar; Marzouki, Mohamed Néjib; Acién-Fernández, Francisco Gabriel; Fernández-Sevilla, José María; Molina-Grima, Emilio; Smaali, Issam

2015-08-01

A coupled process combining microalgae production with direct supercritical biodiesel conversion using a reduced number of operating steps is proposed in this work. Two newly isolated native microalgae strains, identified as Chlorella sp. and Nannochloris sp., were cultivated in both batch and continuous modes. Maximum productivities were achieved during continuous cultures with 318mg/lday and 256mg/lday for Chlorella sp. and Nannochloris sp., respectively. Microalgae were further characterized by determining their photosynthetic performance and nutrient removal efficiency. Biodiesel was produced by catalyst-free in situ supercritical methanol transesterification of wet unwashed algal biomass (75wt.% of moisture). Maximum biodiesel yields of 45.62wt.% and 21.79wt.% were reached for Chlorella sp. and Nannochloris sp., respectively. The analysis of polyunsaturated fatty acids of Chlorella sp. showed a decrease in their proportion when comparing conventional and supercritical transesterification processes (from 37.4% to 13.9%, respectively), thus improving the quality of the biodiesel. Copyright © 2015 Elsevier Ltd. All rights reserved.

A modified efficient method for dental pulp stem cell isolation.

Science.gov (United States)

Raoof, Maryam; Yaghoobi, Mohammad Mehdi; Derakhshani, Ali; Kamal-Abadi, Ali Mohammadi; Ebrahimi, Behnam; Abbasnejad, Mehdi; Shokouhinejad, Noushin

2014-03-01

Dental pulp stem cells can be used in regenerative endodontic therapy. The aim of this study was to introduce an efficient method for dental pulp stem cells isolation. In this in-vitro study, 60 extracted human third molars were split and pulp tissue was extracted. Dental pulp stem cells were isolated by the following three different methods: (1) digestion of pulp by collagenase/dispase enzyme and culture of the released cells; (2) outgrowth of the cells by culture of undigested pulp pieces; (3) digestion of pulp tissue pieces and fixing them. The cells were cultured in minimum essential medium alpha modification (αMEM) medium supplemented with 20% fetal bovine serum(FBS) in humid 37°C incubator with 5% CO 2. The markers of stem cells were studied by reverse transcriptase polymerase chain reaction (PCR). The student t-test was used for comparing the means of independent groups. P third method, we obtained stem cells successfully with about 60% efficiency after 2 days. The results of RT-PCR suggested the expression of Nanog, Oct-4, and Nucleostemin markers in the isolated cells from dental pulps. This study proposes a new method with high efficacy to obtain dental pulp stem cells in a short time.

Method for the detection and isolation of traces of organic fluorine compounds in plants

Energy Technology Data Exchange (ETDEWEB)

Wade, R H; Ross, J M; Benedict, H M

1964-01-01

A method for the detection and isolation of sub-microgram quantities of organic fluorine compounds from plant materials in the presence of much larger amounts of inorganic fluoride is presented. The procedure consists first of a rapid screening step for use with large numbers of vegetable samples and extracts and, second, of a chromatographic step to isolate and characterize any fluoro-organics found. These methods are developed in light of specific chemical characteristics of organic fluorine compounds as a general class. A modification of SOEP's quantitative sub-micro fluoride analytical method is presented as applicable to these isolation methods. Microgram quantities of organic fluorine compounds were found in the plant materials investigated but at a level too low for isolation and identification.

A simple method for human peripheral blood monocyte Isolation

Directory of Open Access Journals (Sweden)

Marcos C de Almeida

2000-04-01

Full Text Available We describe a simple method using percoll gradient for isolation of highly enriched human monocytes. High numbers of fully functional cells are obtained from whole blood or buffy coat cells. The use of simple laboratory equipment and a relatively cheap reagent makes the described method a convenient approach to obtaining human monocytes.

Comparison of in Situ and ex Situ Methods for Synthesis of Two-Photon Polymerization Polymer Nanocomposites

Directory of Open Access Journals (Sweden)

Qingchuan Guo

2014-07-01

Full Text Available This article reports about nanocomposites, which refractive index is tuned by adding TiO2 nanoparticles. We compare in situ/ex situ preparation of nanocomposites. Preparation procedure is described, properties of nanocomposites are compared, and especially we examine the applicability of two-photon polymerization (2PP of synthesized nanocomposites. All prepared samples exhibit suitable optical transparency at specific laser wavelengths. Three-dimensional structures were generated by means of two-photon polymerization effect induced by a femtosecond laser.

Review of the ISOL Method

CERN Document Server

Lindroos, M

2004-01-01

The ISOL technique was invented in Copenhagen over 50 years ago and eventually migrated to CERN where a suitable proton drive beam was available at the Syncho-Cyclotron. The quick spread of the technique from CERN to many other laboratories has resulted in a large user community, which has assured the continued development of the method, physics in the front-line of fundamental research and the application of the method to many applied sciences. The technique is today established as one of the main techniques for on-line isotope production of high intensity and high quality beams. The thick targets used allows the production of unmatched high intensity radioactive beams. The fact that the ions are produced at rest makes it ideally suitable for low energy experiments and for post acceleration using well established accelerator techniques. The many different versions of the technique will be discussed and the many facilities spread all over the world will be reviewed. The major developments at the existing faci...

Matrix diffusion studies by electrical conductivity methods. Comparison between laboratory and in-situ measurements

International Nuclear Information System (INIS)

Ohlsson, Y.; Neretnieks, I.

1998-01-01

Traditional laboratory diffusion experiments in rock material are time consuming, and quite small samples are generally used. Electrical conductivity measurements, on the other hand, provide a fast means for examining transport properties in rock and allow measurements on larger samples as well. Laboratory measurements using electrical conductivity give results that compare well to those from traditional diffusion experiments. The measurement of the electrical resistivity in the rock surrounding a borehole is a standard method for the detection of water conducting fractures. If these data could be correlated to matrix diffusion properties, in-situ diffusion data from large areas could be obtained. This would be valuable because it would make it possible to obtain data very early in future investigations of potentially suitable sites for a repository. This study compares laboratory electrical conductivity measurements with in-situ resistivity measurements from a borehole at Aespoe. The laboratory samples consist mainly of Aespoe diorite and fine-grained granite and the rock surrounding the borehole of Aespoe diorite, Smaaland granite and fine-grained granite. The comparison shows good agreement between laboratory measurements and in-situ data

A combined dynamic analysis method for geometrically nonlinear vibration isolators with elastic rings

Science.gov (United States)

Hu, Zhan; Zheng, Gangtie

2016-08-01

A combined analysis method is developed in the present paper for studying the dynamic properties of a type of geometrically nonlinear vibration isolator, which is composed of push-pull configuration rings. This method combines the geometrically nonlinear theory of curved beams and the Harmonic Balance Method to overcome the difficulty in calculating the vibration and vibration transmissibility under large deformations of the ring structure. Using the proposed method, nonlinear dynamic behaviors of this isolator, such as the lock situation due to the coulomb damping and the usual jump resulting from the nonlinear stiffness, can be investigated. Numerical solutions based on the primary harmonic balance are first verified by direct integration results. Then, the whole procedure of this combined analysis method is demonstrated and validated by slowly sinusoidal sweeping experiments with different amplitudes of the base excitation. Both numerical and experimental results indicate that this type of isolator behaves as a hardening spring with increasing amplitude of the base excitation, which makes it suitable for isolating both steady-state vibrations and transient shocks.

Specific single-cell isolation and genomic amplification of uncultured microorganisms

DEFF Research Database (Denmark)

Kvist, Thomas; Ahring, Birgitte Kiær; Lasken, R.S.

2007-01-01

We in this study describe a new method for genomic studies of individual uncultured prokaryotic organisms, which was used for the isolation and partial genome sequencing of a soil archaeon. The diversity of Archaea in a soil sample was mapped by generating a clone library using group-specific pri......We in this study describe a new method for genomic studies of individual uncultured prokaryotic organisms, which was used for the isolation and partial genome sequencing of a soil archaeon. The diversity of Archaea in a soil sample was mapped by generating a clone library using group......-specific primers in combination with a terminal restriction fragment length polymorphism profile. Intact cells were extracted from the environmental sample, and fluorescent in situ hybridization probing with Cy3-labeled probes designed from the clone library was subsequently used to detect the organisms...... of interest. Single cells with a bright fluorescent signal were isolated using a micromanipulator and the genome of the single isolated cells served as a template for multiple displacement amplification (MDA) using the Phi29 DNA polymerase. The generated MDA product was afterwards used for 16S rRNA gene...

The SENSEI Generic In Situ Interface

Energy Technology Data Exchange (ETDEWEB)

Ayachit, Utkarsh [Kitware, Inc., Clifton Park, NY (United States); Whitlock, Brad [Intelligent Light, Rutherford, NJ (United States); Wolf, Matthew [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Loring, Burlen [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Geveci, Berk [Kitware, Inc., Clifton Park, NY (United States); Lonie, David [Kitware, Inc., Clifton Park, NY (United States); Bethel, E. Wes [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2017-04-11

The SENSEI generic in situ interface is an API that promotes code portability and reusability. From the simulation view, a developer can instrument their code with the SENSEI API and then make make use of any number of in situ infrastructures. From the method view, a developer can write an in situ method using the SENSEI API, then expect it to run in any number of in situ infrastructures, or be invoked directly from a simulation code, with little or no modification. This paper presents the design principles underlying the SENSEI generic interface, along with some simplified coding examples.

Calibration of HPGe detector for in situ measurements of 137Cs in soil by 'peak to valley' method

International Nuclear Information System (INIS)

Fueloep, M.

2000-01-01

The contamination of soil with gamma-ray emitters can be measured in two ways: soil sampling method and in situ spectrometry of the ambient gamma-ray radiation. The conventional soil sampling method has two disadvantages: samples may not be representative for a large areas and determination of the depth distribution of radionuclide requires the measurement of several samples taken from different depths. In situ measurement of a radionuclide activity in soil is more sensitive and provides more representative data than data obtained by soil sample collection and subsequent laboratory analysis. In emergency situations time to assess the contamination is critical. For rapid assessment of the deposited activity direct measurement of ambient gamma-ray radiation are used. In order to obtain accurate measurements of radionuclides in the soil, the detector should be placed on relatively even and open terrain. It is our customary practice to place the detector 1 m above the soil surface. At this height, a tripod-mounted detector can be handled easily and still provide a radius of view for gamma emitting sources out to about 10 m. The 'field of view' actually varies, being somewhat larger for higher sources. Depending upon source energy, the detector effectively sees down to a depth of 15-30 cm. Commonly used method for field gamma spectrometry is method by Beck (1). The most important disadvantages of in situ spectrometry by Beck are that the accuracy of the analysis depends on a separate knowledge of the radioactivity distribution with soil depth. This information can be obtained by calculations using data from in situ measurements and energy dependence of absorption and scattering of photons in soil and track length distribution of photons in soil (2). A method of in situ measurements of 137 Cs in soil where radionuclide distribution in soil profile is calculated by unfolding of detector responses in the full energy peak net area at 0.662 MeV and in the valley under the

High-gravity combustion synthesis and in situ melt infiltration: A new method for preparing cemented carbides

International Nuclear Information System (INIS)

Liu, Guanghua; Li, Jiangtao; Yang, Zengchao; Guo, Shibin; Chen, Yixiang

2013-01-01

A new method of high-gravity combustion synthesis and in situ melt infiltration is reported for preparing cemented carbides, where hot nickel melt is in situ synthesized from a highly exothermic combustion reaction and then infiltrated into tungsten carbide powder compacts. The as-prepared sample showed a homogeneous microstructure, and its relative density, hardness and flexural strength were 94.4%, 84 HRA and 1.49 GPa, respectively. Compared with conventional powder metallurgy approaches, high-gravity combustion synthesis offers a fast and furnace-free way to produce cemented carbides

WIPP R and D in situ test program

International Nuclear Information System (INIS)

Tyler, L.D.

1987-01-01

The Waste Isolation Pilot Plant (WIPP) is a Department of Energy (DOE) RandD Facility for the purpose of developing the technology needed for the safe disposal of the United States' defense-related radioactive waste. The in situ test program focus is to provide the models and data to demonstrate the facility performance for isolation of waste at WIPP. The program is defined for the WIPP sealing system, thermal-structural interactions and waste package performance. A number of integrated large-scale underground tests have been operational since 1983 and are ongoing. The tests address the issues of both systems design and long-term isolation performance of the WIPP repository

In-situ short-circuit protection system and method for high-energy electrochemical cells

Science.gov (United States)

Gauthier, Michel; Domroese, Michael K.; Hoffman, Joseph A.; Lindeman, David D.; Noel, Joseph-Robert-Gaetan; Radewald, Vern E.; Rouillard, Jean; Rouillard, Roger; Shiota, Toshimi; Trice, Jennifer L.

2003-04-15

An in-situ thermal management system for an energy storage device. The energy storage device includes a plurality of energy storage cells each being coupled in parallel to common positive and negative connections. Each of the energy storage cells, in accordance with the cell's technology, dimensions, and thermal/electrical properties, is configured to have a ratio of energy content-to-contact surface area such that thermal energy produced by a short-circuit in a particular cell is conducted to a cell adjacent the particular cell so as to prevent the temperature of the particular cell from exceeding a breakdown temperature. In one embodiment, a fuse is coupled in series with each of a number of energy storage cells. The fuses are activated by a current spike capacitively produced by a cell upon occurrence of a short-circuit in the cell, thereby electrically isolating the short-circuited cell from the common positive and negative connections.

In-situ short circuit protection system and method for high-energy electrochemical cells

Science.gov (United States)

Gauthier, Michel; Domroese, Michael K.; Hoffman, Joseph A.; Lindeman, David D.; Noel, Joseph-Robert-Gaetan; Radewald, Vern E.; Rouillard, Jean; Rouillard, Roger; Shiota, Toshimi; Trice, Jennifer L.

2000-01-01

An in-situ thermal management system for an energy storage device. The energy storage device includes a plurality of energy storage cells each being coupled in parallel to common positive and negative connections. Each of the energy storage cells, in accordance with the cell's technology, dimensions, and thermal/electrical properties, is configured to have a ratio of energy content-to-contact surface area such that thermal energy produced by a short-circuit in a particular cell is conducted to a cell adjacent the particular cell so as to prevent the temperature of the particular cell from exceeding a breakdown temperature. In one embodiment, a fuse is coupled in series with each of a number of energy storage cells. The fuses are activated by a current spike capacitively produced by a cell upon occurrence of a short-circuit in the cell, thereby electrically isolating the short-circuited cell from the common positive and negative connections.

Experimental methods for in situ studies of morphology development during flow

DEFF Research Database (Denmark)

Lyngaae-Jørgensen, Jørgen

1996-01-01

A short overview of in situ methods is given. Multiple laminates of thin films is believed to represent unstable transition structures in many processing operations. A hypotesis for the break up is that first thin cylinders are formed . At a later stage these cylinders break up to spheres...... breakup mechanism involving creation of parallel layers of thin, filmlike formations parallel with the original film is observed. These layers then break up to a final state encompasing nearly spherical drops spread over a thickness much larger (by orders of magnitude) than the original film thickness....

A modified efficient method for dental pulp stem cell isolation

Directory of Open Access Journals (Sweden)

Maryam Raoof

2014-01-01

Full Text Available Background: Dental pulp stem cells can be used in regenerative endodontic therapy. The aim of this study was to introduce an efficient method for dental pulp stem cells isolation. Materials and Methods: In this in-vitro study, 60 extracted human third molars were split and pulp tissue was extracted. Dental pulp stem cells were isolated by the following three different methods: (1 digestion of pulp by collagenase/dispase enzyme and culture of the released cells; (2 outgrowth of the cells by culture of undigested pulp pieces; (3 digestion of pulp tissue pieces and fixing them. The cells were cultured in minimum essential medium alpha modification (αMEM medium supplemented with 20% fetal bovine serum(FBS in humid 37°C incubator with 5% CO 2 . The markers of stem cells were studied by reverse transcriptase polymerase chain reaction (PCR. The student t-test was used for comparing the means of independent groups. P <0.05 was considered as significant. Results: The results indicated that by the first method a few cell colonies with homogenous morphology were detectable after 4 days, while in the outgrowth method more time was needed (10-12 days to allow sufficient numbers of heterogeneous phenotype stem cells to migrate out of tissue. Interestingly, with the improved third method, we obtained stem cells successfully with about 60% efficiency after 2 days. The results of RT-PCR suggested the expression of Nanog, Oct-4, and Nucleostemin markers in the isolated cells from dental pulps. Conclusion: This study proposes a new method with high efficacy to obtain dental pulp stem cells in a short time.

Reducing Water Sensitivity of Chitosan Biocomposite Films Using Gliadin Particles Made by In Situ Method

Directory of Open Access Journals (Sweden)

Dajian Huang

2017-11-01

Full Text Available In order to sustain rapid expansion in the field of biocomposites, it is necessary to develop novel fillers that are biodegradable, and easy to disperse and obtain. In this work, gliadin particles (GPs fabricated through an in situ method have been reported as fillers for creating chitosan (CS-based biocomposite films. In general, the particles tend to agglomerate in the polymer matrix at high loading (approximately >10% in the biopolymer/particles composites prepared by the traditional solution-blending method. However, the micrographs of biocomposites confirmed that the GPs are well dispersed in the CS matrix in all CS/GPs composites even at a high loading of 30% in this study. It was found that the GPs could improve the mechanical properties of the biocomposites. In addition, the results of moisture uptake and solubility in water of biocomposites showed that water resistance of biocomposites was enhanced by the introduction of GPs. These results suggested that GPs fabricated through an in situ method could be a good candidate for use in biopolymer-based composites.

Development of an evaluation method for seismic isolation systems of nuclear power facilities. Seismic design analysis methods for crossover piping system

International Nuclear Information System (INIS)

Tai, Koichi; Sasajima, Keisuke; Fukushima, Shunsuke; Takamura, Noriyuki; Onishi, Shigenobu

2014-01-01

This paper provides seismic design analysis methods suitable for crossover piping system, which connects between seismic isolated building and non-isolated building in the seismic isolated nuclear power plant. Through the numerical study focused on the main steam crossover piping system, seismic response spectrum analysis applying ISM (Independent Support Motion) method with SRSS combination or CCFS (Cross-oscillator, Cross-Floor response Spectrum) method has found to be quite effective for the seismic design of multiply supported crossover piping system. (author)

Novel in situ radiotracer methods for the direct and indirect study of chromate adsorption on alumina

International Nuclear Information System (INIS)

Gancs, L.; Nemeth, Z.; Horanyi, G.

2002-01-01

Radiotracer methods, particularly the radiotracer thin foil method, provide unique possibility of in situ monitoring of chromate adsorption on powdered adsorbents. Two different versions of the thin foil method can be distinguished. In the direct method, the species to be studied is labelled and the radiation measured gives direct information on the adsorption of this species. In the indirect method, a different labelled indicator species is added to the system and the adsorption of this species is followed and the adsorption of the species to be studied is determined based on analysis of the competitive adsorption processes. Both methods were used in the present study. In the in situ methods, the radiation measured consists of two main parts, one coming from the solution background, the other originating from the adsorption layer. In the case of the thin foil method using isotopes emitting soft β - radiation or low energy X-ray the solution background is governed and minimised by self-absorption of the radiation. In the direct study we applied an experimental methodology based on the energy selective measurement of the characteristic K α,β X-radiation emitted by the 51 Cr-labelled chromate species, whereas 35 S-labelled sulphate ions were used as the indicator species in the indirect study. (P.A.)

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

Directory of Open Access Journals (Sweden)

Pirttila A. M.

2009-04-01

Full Text Available A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290 which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH. The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the plantlets infected with M. radiotolerans IMBG290.

A new in-situ method to determine the apparent gas diffusion coefficient of soils

Science.gov (United States)

Laemmel, Thomas; Paulus, Sinikka; Schack-Kirchner, Helmer; Maier, Martin

2015-04-01

Soil aeration is an important factor for the biological activity in the soil and soil respiration. Generally, gas exchange between soil and atmosphere is assumed to be governed by diffusion and Fick's Law is used to describe the fluxes in the soil. The "apparent soil gas diffusion coefficient" represents the proportional factor between the flux and the gas concentration gradient in the soil and reflects the ability of the soil to "transport passively" gases through the soil. One common way to determine this coefficient is to take core samples in the field and determine it in the lab. Unfortunately this method is destructive and needs laborious field work and can only reflect a small fraction of the whole soil. As a consequence insecurity about the resulting effective diffusivity on the profile scale must remain. We developed a new in-situ method using new gas sampling device, tracer gas and inverse soil gas modelling. The gas sampling device contains several sampling depths and can be easily installed into vertical holes of an auger, which allows for fast installation of the system. At the lower end of the device inert tracer gas is injected continuously. The tracer gas diffuses into the surrounding soil. The resulting distribution of the tracer gas concentrations is used to deduce the diffusivity profile of the soil. For Finite Element Modeling of the gas sampling device/soil system the program COMSOL is used. We will present the results of a field campaign comparing the new in-situ method with lab measurements on soil cores. The new sampling pole has several interesting advantages: it can be used in-situ and over a long time; so it allows following modifications of diffusion coefficients in interaction with rain but also vegetation cycle and wind.

In-situ testing methods using radioactive tracers

International Nuclear Information System (INIS)

Sauzay, G.

1976-01-01

Some typical applications of tracer techniques in hydrology are presented: study of the extraction of sands and gravels in a estuary; in-situ study of the transport of sediments by the swell at a depth ranging from 8 to 22m; study of the transport of sands on the site Bonne Anse - Saint Palais [fr

Biofilm biodiversity presented by fluorescent in situ hybridisation

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Wolf Mirela

2017-01-01

Full Text Available Numerous microorganisms may be present in the water distribution system. This is associated with the imperfection of purification processes, or secondary water pollution. Not only it results in the deterioration of water quality parameters, but it also increases threat of epidemiological problems. The water that is biologically unstable creates ideal conditions for colonization of the microorganisms to the inner surface of pipelines which may form biofilm. The key issue, enabling prevention and control of the impact of the development of biofilms, is to assess the biodiversity of microbiocenosis. In order to obtain comprehensive characteristics of microorganisms communities on a particular substrate, it is necessary to combine several techniques. Further analysis using molecular biology methods are usually after traditional methods of assessing the microbiological quality of water. Standard methods do not reflect the actual species composition, because they are targeted at the bacteria that can be isolated and cultured in the laboratory. Conventional methods are capable of detecting less than 10% of the organisms in the sample. In order to study the biodiversity of organisms inhabiting a biofilm (apart from the conventional methods analyses of the diversity of nucleic acids should be used. The first method could be the polymerase chain reaction (PCR and denaturing gradient gel electrophoresis (DGGE. Another way may be fluorescence in situ hybridization, which allows to detect determined DNA sequence using specially labeled oligonucleotide probes. Visualization of the material is performed using a fluorescence microscope. The main purpose of this article is to present rapid and precise identification groups of microorganisms in their natural habitat in biofilm using fluorescent in situ hybridization method (FISH . FISH method can be successfully used to visualize these microorganisms, which show difficulties in culturing, as well as to provide

Combining in situ characterization methods in one set-up: looking with more eyes into the intricate chemistry of the synthesis and working of heterogeneous catalysts.

Science.gov (United States)

Bentrup, Ursula

2010-12-01

Several in situ techniques are known which allow investigations of catalysts and catalytic reactions under real reaction conditions using different spectroscopic and X-ray methods. In recent years, specific set-ups have been established which combine two or more in situ methods in order to get a more detailed understanding of catalytic systems. This tutorial review will give a summary of currently available set-ups equipped with multiple techniques for in situ catalyst characterization, catalyst preparation, and reaction monitoring. Besides experimental and technical aspects of method coupling including X-ray techniques, spectroscopic methods (Raman, UV-vis, FTIR), and magnetic resonance spectroscopies (NMR, EPR), essential results will be presented to demonstrate the added value of multitechnique in situ approaches. A special section is focussed on selected examples of use which show new developments and application fields.

Absorption mechanism of three curcumin constituents through in situ intestinal perfusion method

Directory of Open Access Journals (Sweden)

Y.-H. Wang

2017-09-01

Full Text Available This study aimed to investigate the absorption mechanism of three curcumin constituents in rat small intestines. Self-emulsification was used to solubilize the three curcumin constituents, and the rat in situ intestinal perfusion method was used to study factors on drug absorption, including drug mass concentration, absorption site, and the different types and concentrations of absorption inhibitors. Within the scope of experimental concentrations, three curcumin constituents were absorbed in rat small intestines through the active transport mechanism.

Non liquid nitrogen-based-method for isolation of DNA from ...

African Journals Online (AJOL)

A simple, efficient, reliable and cost-effective method for isolation of total genomic DNA from fungi, suitable for polymerase chain reaction (PCR) amplification and other molecular applications was described. The main advantages of the method are: (1) does not require the use of liquid nitrogen for preparation of fungi DNA; ...

Case studies illustrating in-situ remediation methods for soil and groundwater contaminated with petrochemicals

Energy Technology Data Exchange (ETDEWEB)

Dixon, Robert A; Lance, P E; Downs, A; Kier, Brian P [EMCON Northwest Inc., Portland, OR (United States)

1994-12-31

Four case studies of successful in-situ remediation are summarized illustrating cost-effective methods to remediate soil and groundwater contaminated with volatile and non-volatile petrochemicals. Each site is in a different geologic environment with varying soil types and with and without groundwater impact. The methods described include vadose zone vapor extraction, high-vacuum vapor extraction combined with groundwater tab.le depression, air sparging with groundwater recovery and vapor extraction, and bio remediation of saturated zone soils using inorganic nutrient and oxygen addition

Case studies illustrating in-situ remediation methods for soil and groundwater contaminated with petrochemicals

Energy Technology Data Exchange (ETDEWEB)

Dixon, Robert A.; Lance, P.E.; Downs, A.; Kier, Brian P. [EMCON Northwest Inc., Portland, OR (United States)

1993-12-31

Four case studies of successful in-situ remediation are summarized illustrating cost-effective methods to remediate soil and groundwater contaminated with volatile and non-volatile petrochemicals. Each site is in a different geologic environment with varying soil types and with and without groundwater impact. The methods described include vadose zone vapor extraction, high-vacuum vapor extraction combined with groundwater tab.le depression, air sparging with groundwater recovery and vapor extraction, and bio remediation of saturated zone soils using inorganic nutrient and oxygen addition

Comparison study for the CCME reference method for determination of PHC in soil by using internal and external standard methods and by using silica gel column cleanup and in-situ silica gel cleanup methods

International Nuclear Information System (INIS)

Wang, Z.; Fingas, M.; Sigouin, L.; Yang, C.; Hollebone, B.

2003-01-01

The assessment, cleanup, and remediation of hydrocarbon contaminated sites is covered in the Reference Method for Canada-Wide Standard for Petroleum Hydrocarbons-Tier 1 Method. It replaces several analytical methods used in the past by some laboratories and jurisdictions in Canada. The authors conducted two comparative evaluations to validate the Tier 1 Analytical Method. The first compared the Internal and External Standard Methods, and the second compared the Silica Gel Column Cleanup Method with the In-situ Silica Gel Cleanup Method. The Canadian Council of Ministers of the Environment (CCME) Tier 1 Method recommends and requires the External Standard Method to determine petroleum hydrocarbons (PHC) in soil samples. The Internal Method is widely used to quantify various organic and inorganic pollutants in environmental samples. The Tier 1 Method offers two options for the same extract cleanup. They are: Option A - In-situ Silica Gel Cleanup, and Option B - Silica Gel Column Cleanup. Linearity, precision, and PHC quantification results were the parameters considered for diesel and motor oil solutions, for diesel spiked soil samples, and for motor oil spiked soil samples. It was concluded that both the External and Internal Standard Methods for gas chromatograph (GC) determination of PHC in soil possess their own advantages. The PHC results obtained using the In-Situ Silica Gel Cleanup Method were lower than those obtained with the Silica Gel Column Cleanup Methods. The more efficient and effective sample cleanup method proved to be the Silica Gel Column Method. 13 refs., 7 tabs., 7 figs

WIPP/SRL in-situ tests

International Nuclear Information System (INIS)

Mamsey, W.G.

1990-01-01

The Materials Interface Interactions Test (MIIT) is the only in-situ program involving the burial of simulated high-level waste forms operating in the United States. Fifteen glass and waste form compositions and their proposed package materials, supplied by 7 countries, are interred in salt at the Waste Isolation Pilot Plant (WIPP) in Carlsbad, New Mexico. A joint effort between Sandia National Laboratories and Savannah River Laboratory, MIIT is the largest international cooperative in-situ venture yet undertaken. The objective of the current study is to document the waste form compositions used in the MIIT program and then to examine compositional correlations based on structural considerations, bonding energies, and surface layer formation. These correlations show important similarities between the many different waste glass compositions studied world wide and suggest that these glasses would be expected to perform well and in a similar manner

In situ electrochemical enrichment and isolation of a magnetite-reducing bacterium from a high pH serpentinizing spring.

Science.gov (United States)

Rowe, Annette R; Yoshimura, Miho; LaRowe, Doug E; Bird, Lina J; Amend, Jan P; Hashimoto, Kazuhito; Nealson, Kenneth H; Okamoto, Akihiro

2017-06-01

Serpentinization is a geologic process that produces highly reduced, hydrogen-rich fluids that support microbial communities under high pH conditions. We investigated the activity of microbes capable of extracellular electron transfer in a terrestrial serpentinizing system known as 'The Cedars'. Measuring current generation with an on-site two-electrode system, we observed daily oscillations in current with the current maxima and minima occurring during daylight hours. Distinct members of the microbial community were enriched. Current generation in lab-scale electrochemical reactors did not oscillate, but was correlated with carbohydrate amendment in Cedars-specific minimal media. Gammaproteobacteria and Firmicutes were consistently enriched from lab electrochemical systems on δ-MnO 2 and amorphous Fe(OH) 3 at pH 11. However, isolation of an electrogenic strain proved difficult as transfer cultures failed to grow after multiple rounds of media transfer. Lowering the bulk pH in the media allowed us to isolate a Firmicutes strain (Paenibacillus sp.). This strain was capable of electrode and mineral reduction (including magnetite) at pH 9. This report provides evidence of the in situ activity of microbes using extracellular substrates as sinks for electrons at The Cedars, but also highlights the potential importance of community dynamics for supporting microbial life through either carbon fixation, and/or moderating pH stress. © 2017 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.

Isolation and HPLC method development of azafrin from Alectra parasitica var. chitrakutensis.

Science.gov (United States)

Agrawal, Poonam; Laddha, Kirti; Tiwari, Ashok

2014-01-01

This study was undertaken to isolate and quantify azafrin in Alectra parasitica (Scrophulariaceae) rhizomes. A simple method for the isolation of carotenoid, azafrin, involves solvent extraction of the dried rhizome powder using a single solvent and further purification by recrystallisation. The structure of the compound was elucidated and confirmed by thin-layer chromatography, infrared spectroscopy, mass spectroscopy and nuclear magnetic resonance spectral analysis. A specific and rapid reversed-phase high-performance liquid chromatography (HPLC) method was developed for the analysis of azafrin. The method was validated for accuracy, precision, linearity and specificity. Validation revealed that the method is specific, accurate, precise, reliable and reproducible. The proposed HPLC method can be used for the identification and quantitative analysis of azafrin in A. parasitica rhizomes.

A simple and efficient method for isolating small RNAs from different plant species

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de Folter Stefan

2011-02-01

Full Text Available Abstract Background Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species. Results We developed a simple and efficient method to isolate small RNAs from different plant species by first comparing different total RNA extraction protocols, followed by streamlining the best one, finally resulting in a small RNA extraction method that has no need of first total RNA extraction and is not based on the commercially available TRIzol® Reagent or columns. This small RNA extraction method not only works well for plant tissues with high polysaccharide content, like cactus, agave, banana, and tomato, but also for plant species like Arabidopsis or tobacco. Furthermore, the obtained small RNA samples were successfully used in northern blot assays. Conclusion Here we provide a simple and efficient method to isolate small RNAs from different plant species, such as cactus, agave, banana, tomato, Arabidopsis, and tobacco, and the small RNAs from this simplified and low cost method is suitable for downstream handling like northern blot assays.

Method for local temperature measurement in a nanoreactor for in situ high-resolution electron microscopy.

Science.gov (United States)

Vendelbo, S B; Kooyman, P J; Creemer, J F; Morana, B; Mele, L; Dona, P; Nelissen, B J; Helveg, S

2013-10-01

In situ high-resolution transmission electron microscopy (TEM) of solids under reactive gas conditions can be facilitated by microelectromechanical system devices called nanoreactors. These nanoreactors are windowed cells containing nanoliter volumes of gas at ambient pressures and elevated temperatures. However, due to the high spatial confinement of the reaction environment, traditional methods for measuring process parameters, such as the local temperature, are difficult to apply. To address this issue, we devise an electron energy loss spectroscopy (EELS) method that probes the local temperature of the reaction volume under inspection by the electron beam. The local gas density, as measured using quantitative EELS, is combined with the inherent relation between gas density and temperature, as described by the ideal gas law, to obtain the local temperature. Using this method we determined the temperature gradient in a nanoreactor in situ, while the average, global temperature was monitored by a traditional measurement of the electrical resistivity of the heater. The local gas temperatures had a maximum of 56 °C deviation from the global heater values under the applied conditions. The local temperatures, obtained with the proposed method, are in good agreement with predictions from an analytical model. Copyright © 2013 Elsevier B.V. All rights reserved.

In-situ uranium leaching

International Nuclear Information System (INIS)

Dotson, B.J.

1986-01-01

This invention provides a method for improving the recovery of mineral values from ore bodies subjected to in-situ leaching by controlling the flow behaviour of the leaching solution. In particular, the invention relates to an in-situ leaching operation employing a foam for mobility control of the leaching solution. A foam bank is either introduced into the ore bed or developed in-situ in the ore bed. The foam then becomes a diverting agent forcing the leaching fluid through the previously non-contacted regions of the deposit

Scalable Analysis Methods and In Situ Infrastructure for Extreme Scale Knowledge Discovery

Energy Technology Data Exchange (ETDEWEB)

Bethel, Wes [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2016-07-24

The primary challenge motivating this team’s work is the widening gap between the ability to compute information and to store it for subsequent analysis. This gap adversely impacts science code teams, who are able to perform analysis only on a small fraction of the data they compute, resulting in the very real likelihood of lost or missed science, when results are computed but not analyzed. Our approach is to perform as much analysis or visualization processing on data while it is still resident in memory, an approach that is known as in situ processing. The idea in situ processing was not new at the time of the start of this effort in 2014, but efforts in that space were largely ad hoc, and there was no concerted effort within the research community that aimed to foster production-quality software tools suitable for use by DOE science projects. In large, our objective was produce and enable use of production-quality in situ methods and infrastructure, at scale, on DOE HPC facilities, though we expected to have impact beyond DOE due to the widespread nature of the challenges, which affect virtually all large-scale computational science efforts. To achieve that objective, we assembled a unique team of researchers consisting of representatives from DOE national laboratories, academia, and industry, and engaged in software technology R&D, as well as engaged in close partnerships with DOE science code teams, to produce software technologies that were shown to run effectively at scale on DOE HPC platforms.

Comparison of the Histopaque-1119 method with the Plasmagel method for separation of blood leukocytes for cytomegalovirus isolation.

Science.gov (United States)

Slifkin, M; Cumbie, R

1992-10-01

Histopaque-1119 (Sigma Chemical Co., St. Louis, Mo.) and Plasmagel (Cellular Products, Inc., Buffalo, N.Y.) were compared as density gradient separation reagents for the separation of polymorphonuclear leukocytes and mononuclear cells from blood from the isolation of cytomegalovirus (CMV). Of 200 peripheral blood specimens examined, CMV was recovered from 51 by both methods. The time of detection of immunofluorescent sites or a cytopathic effect associated with CMV was similar by each method. The Histopaque-1119 method was less time-consuming than the Plasmagel method since it did not require a precentrifugation step for the settling of erythrocytes. The use of Histopaque-1119 will permit an effective alternative single-step method for the separation of blood leukocytes for the isolation of CMV.

Extraction Methods for the Isolation of Isoflavonoids from Plant Material

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Blicharski Tomasz

2017-03-01

Full Text Available The purpose of this review is to describe and compare selected traditional and modern extraction methods employed in the isolation of isoflavonoids from plants. Conventional methods such as maceration, percolation, or Soxhlet extraction are still frequently used in phytochemical analysis. Despite their flexibility, traditional extraction techniques have significant drawbacks, including the need for a significant investment of time, energy, and starting material, and a requirement for large amounts of potentially toxic solvents. Moreover, these techniques are difficult to automate, produce considerable amount of waste and pose a risk of degradation of thermolabile compounds. Modern extraction methods, such as: ultrasound-assisted extraction, microwave-assisted extraction, accelerated solvent extraction, supercritical fluid extraction, and negative pressure cavitation extraction, can be regarded as remedies for the aforementioned problems. This manuscript discusses the use of the most relevant extraction techniques in the process of isolation of isoflavonoids, secondary metabolites that have been found to have a plethora of biological and pharmacological activities.

In-situ Crystallization of Highly Volatile Commercial Mold Flux Using an Isolated Observation System in the Confocal Laser Scanning Microscope

Science.gov (United States)

Park, Jun-Yong; Ryu, Jae Wook; Sohn, Il

2014-08-01

The in situ crystallization behavior of highly volatile commercial mold fluxes for medium carbon steels was investigated using the confocal laser scanning microscope (CLSM) equipped with an optimized isolated observation system. The highly volatile compounds of the mold flux were suppressed during heating allowing direct observation in the CLSM. Cooling rates of 25, 50, 100, 400, and 800 K/min were incorporated and continuous cooling transformation (CCT) diagrams of 4 different commercial mold fluxes for medium carbon steels were developed. Identification of the crystalline phase was conducted with XRD and SEM-EDS analysis. A cuspidine crystalline was observed in all samples at various cooling rates. With higher basicity, CaF2, and NaF, the crystallization of the fluxes was enhanced according to the CCT diagram. As the slag structure becomes depolymerized, the diffusion rate of the cathodic ions seems to increase.

A simple method for DNA isolation from Xanthomonas spp.

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Gomes Luiz Humberto

2000-01-01

Full Text Available A simple DNA isolation method was developed with routine chemicals that yields high quality and integrity preparations when compared to some of the most well known protocols. The method described does not require the use of lysing enzymes, water bath and the DNA was obtained within 40 minutes The amount of nucleic acid extracted (measured in terms of absorbancy at 260 nm from strains of Xanthomonas spp., Pseudomonas spp. and Erwinia spp. was two to five times higher than that of the most commonly used method.

Simple and efficient methods for isolation and activity measurement ...

African Journals Online (AJOL)

Jane

2011-06-29

Jun 29, 2011 ... Key words: Hirudin, thrombin titration method, chromatography, purification. INTRODUCTION. Since recombinant ... Escherichia coli in 1986, intensive research had been .... mixed with 50 µl sample was incubated in 37°C water for 5 min, then 5 µl .... conclusion, the concise and efficient isolation line of the.

The method for the in-situ leaching of a uranium mining

International Nuclear Information System (INIS)

Chen Zhen; Xu Xianyi; Wang Xuemin

2011-01-01

The paper reviews the main factors of in-situ leaching for uranium mining. A kind of technique called dilution with few reagent is put forward to the in-situ leaching of sandstone-type uranium deposit with high TDS. This technique can not only effectively prevent the pipe plug, but also can improve the economic benefits. (authors)

Optimization of One-Step In Situ Transesterification Method for Accurate Quantification of EPA in Nannochloropsis gaditana

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Yuting Tang

2016-11-01

Full Text Available Microalgae are a valuable source of lipid feedstocks for biodiesel and valuable omega-3 fatty acids. Nannochloropsis gaditana has emerged as a promising producer of eicosapentaenoic acid (EPA due to its fast growth rate and high EPA content. In the present study, the fatty acid profile of Nannochloropsis gaditana was found to be naturally high in EPA and devoid of docosahexaenoic acid (DHA, thereby providing an opportunity to maximize the efficacy of EPA production. Using an optimized one-step in situ transesterification method (methanol:biomass = 90 mL/g; HCl 5% by vol.; 70 °C; 1.5 h, the maximum fatty acid methyl ester (FAME yield of Nannochloropsis gaditana cultivated under rich condition was quantified as 10.04% ± 0.08% by weight with EPA-yields as high as 4.02% ± 0.17% based on dry biomass. The total FAME and EPA yields were 1.58- and 1.23-fold higher separately than that obtained using conventional two-step method (solvent system: methanol and chloroform. This one-step in situ method provides a fast and simple method to measure fatty acid methyl ester (FAME yields and could serve as a promising method to generate eicosapentaenoic acid methyl ester from microalgae.

Visual and efficient immunosensor technique for advancing biomedical applications of quantum dots on Salmonella detection and isolation

Science.gov (United States)

Tang, Feng; Pang, Dai-Wen; Chen, Zhi; Shao, Jian-Bo; Xiong, Ling-Hong; Xiang, Yan-Ping; Xiong, Yan; Wu, Kai; Ai, Hong-Wu; Zhang, Hui; Zheng, Xiao-Li; Lv, Jing-Rui; Liu, Wei-Yong; Hu, Hong-Bing; Mei, Hong; Zhang, Zhen; Sun, Hong; Xiang, Yun; Sun, Zi-Yong

2016-02-01

It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here by applying fluorescent nanobioprobes on a specially-designed cellulose-based swab (a solid-phase enrichment system). The selective and chromogenic medium used on this swab can achieve the ultrasensitive amplification of target bacteria and form chromogenic colonies in situ based on a simple biochemical reaction. More importantly, because this swab can serve as an attachment site for the targeted pathogens to immobilize and immunologically capture nanobioprobes, our mAb-conjugated QD bioprobes were successfully applied on the solid-phase enrichment system to capture the fluorescence of targeted colonies under a designed excitation light instrument based on blue light-emitting diodes combined with stereomicroscopy or laser scanning confocal microscopy. Compared with the traditional methods using 4-7 days to isolate Salmonella from the bacterial mixture, this method took only 2 days to do this, and the process of initial screening and preliminary diagnosis can be completed in only one and a half days. Furthermore, the limit of detection can reach as low as 101 cells per mL Salmonella on the background of 105 cells per mL non-Salmonella (Escherichia coli, Proteus mirabilis or Citrobacter freundii, respectively) in experimental samples, and even in human anal ones. The visual and efficient immunosensor technique may be proved to be a favorable alternative for screening and isolating Salmonella in a large number of samples related to public health surveillance.It is a great challenge in nanotechnology for fluorescent nanobioprobes to be applied to visually detect and directly isolate pathogens in situ. A novel and visual immunosensor technique for efficient detection and isolation of Salmonella was established here

Isolated Liver Perfusion Using Percutaneous Methods:[ql An Experimental Study in the Pig

International Nuclear Information System (INIS)

Harnek, Jan; Cwikiel, Wojciech; Bergqvist, Lennart; Persson, Bo; Stridbeck, Hans

1996-01-01

Purpose: To develop a method for isolated perfusion of the liver using radiological methods. Methods: Twenty-one pigs, weighing about 20 kg, were divided into three groups. By transjugular and transfemoral approaches two occlusion balloons were placed in the inferior vena cava cranial and caudal, respectively, to the origin of the hepatic veins. One occlusion balloon was placed transfemorally in the common hepatic artery. Another occlusion balloon was inserted in the main branch of the portal vein via the transjugular-transhepatic approach in 11 pigs (groups 1 and 2), and in 10 pigs (group 3) by a percutaneous transhepatic route. After inflation of the balloons, patency of the isolated liver circulation was evaluated by recirculation of 99 Tc m -labelled human albumin during 30 min. Blood tests were obtained after 1, 3, 5, 10, 15, and 30 min to evaluate leakage from the liver to the systemic circulation. Results: Increasing leakage to the systemic circulation from the isolated liver circulation was observed in groups 1 and 2. In the third group the leakage was less than 10%. Conclusion: In an experimental animal model, isolated perfusion of the liver with minor leakage to the systemic circulation may be achieved using radiological methods

Experimental and mathematical simulation techniques for determining an in-situ response testing method for neutron sensors used in reactor power plant protection systems

International Nuclear Information System (INIS)

Behbahani, A.

1983-01-01

An analytical neutron sensor response model and methods for transient response measurements of neutron sensors (compensated ionization chamber), including possible in-situ techniques have been developed and evaluated to meet the provisions of Draft Standard ISA Sd67.06, IEEE 338-1977, and NRC Guide 1.118. One in-situ method requires the perturbation of the high voltage detector (sensor) power supply and measurement of the sensor response. The response to a perturbation of the power supply is related to the response of the sensor to a transient change in neutron flux. Random signal analysis is another in-situ technique to monitor the neutron sensor response. In this method the power spectrum of the inherent fluctuations from the neutron sensor output (current in CIC) are measured and evaluated. Transient response techniques (including in-situ methods) are experimentally and analytically evaluated to identify the mechanisms which may cause degradation in the response of the neutron sensors. The objective of the experimental evaluation was to correlate the measured response time using transient radiation flux changes and power supply perturbation. The objective of the analytical model of the different sensor response was to predict response time and degradation mechanisms

In Situ Electron Microscopy of Lactomicroselenium Particles in Probiotic Bacteria

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Gabor Nagy

2016-06-01

Full Text Available Electron microscopy was used to test whether or not (a in statu nascendi synthesized, and in situ measured, nanoparticle size does not differ significantly from the size of nanoparticles after their purification; and (b the generation of selenium is detrimental to the bacterial strains that produce them. Elemental nano-sized selenium produced by probiotic latic acid bacteria was used as a lactomicroselenium (lactomicroSel inhibitor of cell growth in the presence of lactomicroSel, and was followed by time-lapse microscopy. The size of lactomicroSel produced by probiotic bacteria was measured in situ and after isolation and purification. For these measurements the TESLA BS 540 transmission electron microscope was converted from analog (aTEM to digital processing (dTEM, and further to remote-access internet electron microscopy (iTEM. Lactobacillus acidophilus produced fewer, but larger, lactomicroSel nanoparticles (200–350 nm than Lactobacillus casei (L. casei, which generated many, smaller lactomicroSel particles (85–200 nm and grains as a cloudy, less electrodense material. Streptococcus thermophilus cells generated selenoparticles (60–280 nm in a suicidic manner. The size determined in situ in lactic acid bacteria was significantly lower than those measured by scanning electron microscopy after the isolation of lactomicroSel particles obtained from lactobacilli (100–500 nm, but higher relative to those isolated from Streptococcus thermopilus (50–100 nm. These differences indicate that smaller lactomicroSel particles could be more toxic to the producing bacteria themselves and discrepancies in size could have implications with respect to the applications of selenium nanoparticles as prebiotics.

Schizosaccharomyces isolation method

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Benito Santiago

2014-01-01

Full Text Available This study discusses the optimization of a selective and differential medium which would facilitate the isolation of Schizosaccharomyces (a genus with a low incidence compared to other microorganisms to select individuals from this genus for industrial purposes, especially in light of the recent recommendation of the use of yeasts from this genus in the wine industry by the International Organisation of Vine and Wine, or to detect the presence of such yeasts, for those many authors who consider them food spoilers. To this end, we studied various selective differential agents based on the main physiological characteristics of these species, such as their high resistances to high concentrations of sugar, sulfur dioxide, sorbic acid, benzoic acid, acetic acid or malo ethanolic fermentation. This selective medium is based on the genus resistance to the antibiotic actidione and its high resistance to inhibitory agents such as benzoic acid. Malic acid was used as a differential factor due to the ability of this genus to metabolise it to ethanol, which allows detecting of the degradation of this compound. Lastly, the medium was successfully used to isolate strains of Schizosaccharomyces pombe from honey and honeycombs.

Developing a data acquisition system for the Waste Isolation Pilot Plant's in situ tests

International Nuclear Information System (INIS)

McIlmoyle, J.T.; Matalucci, R.V.; Ogden, H.C.

1986-01-01

The US Department of Energy is developing the Waste Isolation Pilot Plant in southeastern New Mexico as an R and D facility to demonstrate the safe disposal of radioactive wastes from US defense programs. The in situ tests associated with this facility (659 m below the surface) address technical issues of thermal/structural interaction, waste package performance, and plugging and sealing. Data from these tests are collected by a Data Acquisition System (DAS) consisting of two distinct entities that use the IEEE-488 bus for communication. Up to 5000 data channels can be recorded every 4 hr from extensometers, stressmeters, thermocouples, closure gages, and other special gages. The DAS consists of a multitasking and multiuser ModComp computer housed in a surface trailer that converts the data to engineering units, stores the data in raw and converted form, and provides quick-look plots and data listings. The other part of the DAS consists of the seven underground remote enclosed instrument stations (sheds) located near each test room and up to 1800 m from the surface trailer that contains the instruments and signal conditioning to operate and monitor the tests. The ModComp system manages data acquisition and supports concurrently 12 on-line terminals and two dial-up access lines for data access, gage diagnostics, data handling and updating, and software development. Data are currently acquired from over 4000 gages with 15,000 defined measurands; the system can be expanded to 65,534 measurands. 4 refs., 7 figs

Comparison of Various Methods for Isolation of Nocardia from Soil

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Masoumeh Rasouli-Nasab

2017-02-01

Full Text Available Background The genus Nocardia is Gram-positive, aerobic filamentous bacilli and saprophytic micro-organisms that can be isolated from freshwater, salt water, dust and decaying vegetation especially the soil. This study aimed to investigate the several media for to determine a suitable culture media with the ability to better for the isolation of Nocardia from soil. Methods In this study, 400 soil samples were collected from different areas from Iran. The soil samples were then cultured on the four culture media such as Humic acid vitamin B agar, Paraffin agar, Sabouraud dextrose agar supplemented whit cycloheximide and carbon-free broth containing paraffin rods and incubated at 35°C. All of culture media investigated every 3 days for a month. Colonies suspicious to Nocardia were stained with Gram-stain, acid-fast and partially acid-fast and evaluated for resistance to lysozyme. Results From 400 soil samples, the number of 62, 10, 28 and 19 strains of Nocardia were isolated by paraffin rods, Humic Acid Vitamin B agar, Paraffin agar and Sabouraud dextrose agar whit cycloheximide, respectively. Most Nocardia strains were isolated using paraffin bait technique. Conclusions Isolation of Nocardia spp. is enhanced by using the paraffin baiting technique that relies on the selective ability of this micro-organism to metabolize paraffin.

Comprehensive validation scheme for in situ fiber optics dissolution method for pharmaceutical drug product testing.

Science.gov (United States)

Mirza, Tahseen; Liu, Qian Julie; Vivilecchia, Richard; Joshi, Yatindra

2009-03-01

There has been a growing interest during the past decade in the use of fiber optics dissolution testing. Use of this novel technology is mainly confined to research and development laboratories. It has not yet emerged as a tool for end product release testing despite its ability to generate in situ results and efficiency improvement. One potential reason may be the lack of clear validation guidelines that can be applied for the assessment of suitability of fiber optics. This article describes a comprehensive validation scheme and development of a reliable, robust, reproducible and cost-effective dissolution test using fiber optics technology. The test was successfully applied for characterizing the dissolution behavior of a 40-mg immediate-release tablet dosage form that is under development at Novartis Pharmaceuticals, East Hanover, New Jersey. The method was validated for the following parameters: linearity, precision, accuracy, specificity, and robustness. In particular, robustness was evaluated in terms of probe sampling depth and probe orientation. The in situ fiber optic method was found to be comparable to the existing manual sampling dissolution method. Finally, the fiber optic dissolution test was successfully performed by different operators on different days, to further enhance the validity of the method. The results demonstrate that the fiber optics technology can be successfully validated for end product dissolution/release testing. (c) 2008 Wiley-Liss, Inc. and the American Pharmacists Association

Full-scale demonstration of EBS construction technology I. Block, pellet and in-situ compaction method

International Nuclear Information System (INIS)

Toguri, Satohito; Asano, Hidekazu; Takao, Hajime; Matsuda, Takeshi; Amemiya, Kiyoshi

2008-01-01

(i) Bentonite Block: Applicability of manufacturing technology of buffer material was verified by manufacturing of full scale bentonite ring which consists of one-eight (1/8) dividing block (Outside Diameter (OD): 2.220 mm H: 300 mm). Density characteristic, dimension and scale effect, which were considered the tunnel environment under transportation, were evaluated. Vacuum suction technology was selected as handling technology for the ring. Hoisting characteristic of vacuum suction technology was presented through evaluation of the mechanical property of buffer material, the friction between blocks, etc. by using a full-scale bentonite ring (OD 2.200 mm, H 300 mm). And design of bentonite block and emplacement equipment were presented in consideration of manufacturability of the block, stability of handling and improvement of emplacement efficiency. (ii) Bentonite Pellet Filling: Basic characteristics such as water penetration, swelling and thermal conductivity of various kinds of bentonite pellet were collected by laboratory scale tests. Applicability of pellet filling technology was evaluated by horizontal filling test using a simulated full-scale drift tunnel (OD 2.200 mm, L 6 m) . Filling density, grain size distribution, etc. were also measured. (iii) In-Situ Compaction of Bentonite: Dynamic compaction method (heavy weight fall method) was selected as in-situ compaction technology. Compacting examination which used a full scale disposal pit (OD 2.360 mm) was carried out. Basic specification of compacting equipment and applicability of in-situ compaction technology were presented. Density, density distribution of buffer material and energy acted on the wall of the pit, were also measured. (author)

Application of tomographics methods for determination of rock failure in situ

Directory of Open Access Journals (Sweden)

Pandula Blažej

1996-03-01

Full Text Available Rock Failure determination closely connects with optimalization of rock disintegration as well excavation of underground openings. The aim of this optimalization is than to reach desired fragmentation during the blasting as well as choose the most suitable excavation technology. Study of rock failure can be very useful to reach this goal. In practice it can be done on the basis of personal technological experiences of blasting operator obtained in long term experiences in situ conditions. Because in many cases that experiences can not be utilised, it is necessary to learn against more precise rock failure evaluation. It can also down the costs of disintegration as well as excavation. The paper deals with results of common geophysical methods and methods of seismic tomography achieved at the hidden rock failure determination in the rock masses of the Ruzin dam during projecting the outlet equipment.

Scalable Analysis Methods and In Situ Infrastructure for Extreme Scale Knowledge Discovery

Energy Technology Data Exchange (ETDEWEB)

Duque, Earl P.N. [J.M. Smith International, LLC, Rutherford, NJ (United States). DBA Intelligent Light; Whitlock, Brad J. [J.M. Smith International, LLC, Rutherford, NJ (United States). DBA Intelligent Light

2017-08-25

High performance computers have for many years been on a trajectory that gives them extraordinary compute power with the addition of more and more compute cores. At the same time, other system parameters such as the amount of memory per core and bandwidth to storage have remained constant or have barely increased. This creates an imbalance in the computer, giving it the ability to compute a lot of data that it cannot reasonably save out due to time and storage constraints. While technologies have been invented to mitigate this problem (burst buffers, etc.), software has been adapting to employ in situ libraries which perform data analysis and visualization on simulation data while it is still resident in memory. This avoids the need to ever have to pay the costs of writing many terabytes of data files. Instead, in situ enables the creation of more concentrated data products such as statistics, plots, and data extracts, which are all far smaller than the full-sized volume data. With the increasing popularity of in situ, multiple in situ infrastructures have been created, each with its own mechanism for integrating with a simulation. To make it easier to instrument a simulation with multiple in situ infrastructures and include custom analysis algorithms, this project created the SENSEI framework.

Four Models of In Situ Simulation

DEFF Research Database (Denmark)

Musaeus, Peter; Krogh, Kristian; Paltved, Charlotte

2014-01-01

Introduction In situ simulation is characterized by being situated in the clinical environment as opposed to the simulation laboratory. But in situ simulation bears a family resemblance to other types of on the job training. We explore a typology of in situ simulation and suggest that there are f......Introduction In situ simulation is characterized by being situated in the clinical environment as opposed to the simulation laboratory. But in situ simulation bears a family resemblance to other types of on the job training. We explore a typology of in situ simulation and suggest...... that there are four fruitful approaches to in situ simulation: (1) In situ simulation informed by reported critical incidents and adverse events from emergency departments (ED) in which team training is about to be conducted to write scenarios. (2) In situ simulation through ethnographic studies at the ED. (3) Using...... the following processes: Transition processes, Action processes and Interpersonal processes. Design and purpose This abstract suggests four approaches to in situ simulation. A pilot study will evaluate the different approaches in two emergency departments in the Central Region of Denmark. Methods The typology...

In-Situ Simulation

DEFF Research Database (Denmark)

Bjerregaard, Anders Thais; Slot, Susanne; Paltved, Charlotte

2015-01-01

, and organisational characteristic. Therefore, it might fail to fully mimic real clinical team processes. Though research on in situ simulation in healthcare is in its infancy, literature is abundant on patient safety and team training1. Patient safety reporting systems that identify risks to patients can improve......Introduction: In situ simulation offers on-site training to healthcare professionals. It refers to a training strategy where simulation technology is integrated into the clinical encounter. Training in the simulation laboratory does not easily tap into situational resources, e.g. individual, team...... patient safety if coupled with training and organisational support. This study explored the use of critical incidents and adverse events reports for in situ simulation and short-term observations were used to create learning objectives and training scenarios. Method: This study used an interventional case...

HER2 testing in the UK: recommendations for breast and gastric in-situ hybridisation methods

LENUS (Irish Health Repository)

Bartlett, J. M. S.

2011-01-01

These guidelines supplement existing guidelines on HER2 testing by immunohistochemistry and in-situ hybridisation(ISH) methods in the UK. They provide a specific focus on aspects of guidance relevant to HER2 ISH testing methods, both fluorescent and chromogenic. They are formulated to give advice on methodology, interpretation and quality control for ISH-based testing of HER2 status in common tumour types, including both breast and gastric tumours. The aim is to ensure that all ISH-based testing is accurate, reliable and timely.

Accuracy of phenotypic methicillin susceptibility methods in the detection of Staphylococcus aureus isolates carrying different SCCmec types

Directory of Open Access Journals (Sweden)

Pricilla DM de Matos

2010-11-01

Full Text Available A total of 138 isolates, 118 methicillin-resistant Staphylococcus aureus (MRSA isolates (staphylococcal cassette chromosome type II, 20 isolates, type III, 39 isolates and type IV, 59 isolates and 20 methicillin-sensitive S. aureus isolates were evaluated by phenotypic methods: cefoxitin and oxacillin disk diffusion (DD, agar dilution (AD, latex agglutination (LA, oxacillin agar screening (OAS and chromogenic agar detection. All methods showed 100% specificity, but only the DD tests presented 100% sensitivity. The sensitivity of the other tests ranged from 82.2% (OAS-98.3% (AD. The LA test showed the second lowest sensitivity (86.4%. The DD test showed high accuracy in the detection of MRSA isolates, but there was low precision in the detection of type IV isolates by the other tests, indicating that the genotypic characteristics of the isolates should be considered.

Accuracy of phenotypic methicillin susceptibility methods in the detection of Staphylococcus aureus isolates carrying different SCCmec types.

Science.gov (United States)

Matos, Pricilla D M de; Schuenck, Ricardo P; Cavalcante, Fernanda S; Caboclo, Roberta M F; Santos, Kátia Regina N dos

2010-11-01

A total of 138 isolates, 118 methicillin-resistant Staphylococcus aureus (MRSA) isolates (staphylococcal cassette chromosome type II, 20 isolates, type III, 39 isolates and type IV, 59 isolates) and 20 methicillin-sensitive S. aureus isolates were evaluated by phenotypic methods: cefoxitin and oxacillin disk diffusion (DD), agar dilution (AD), latex agglutination (LA), oxacillin agar screening (OAS) and chromogenic agar detection. All methods showed 100% specificity, but only the DD tests presented 100% sensitivity. The sensitivity of the other tests ranged from 82.2% (OAS)-98.3% (AD). The LA test showed the second lowest sensitivity (86.4%). The DD test showed high accuracy in the detection of MRSA isolates, but there was low precision in the detection of type IV isolates by the other tests, indicating that the genotypic characteristics of the isolates should be considered.

Methods Development for the Isolation and Culture of Primary Corneal Endothelial Cells

Science.gov (United States)

2017-02-01

a cell population particularly suitable for low serum propagation, provided that appropriate growth factors are available. A low serum medium...of MGK. 15. SUBJECT TERMS Cornea, chemical warfare agent, corneal endothelial cell, endothelium, growth , isolation, mouse, rabbit, porcine, in...with corneal SM exposure.2 A primary requirement in achieving this goal is to develop methods that enable the isolation of a pure CEC population and

''In situ'' electronic testing method of a neutron detector performance

International Nuclear Information System (INIS)

Gonzalez, J.M.; Levai, F.

1987-01-01

The method allows detection of any important change in the electrical characteristics of a neutron sensor channel. It checks the response signal produced by an electronic detector circuit when a pulse generator is connected as input signal in the high voltage supply. The electronic circuit compares the detector capacitance value, previously measured, against a reference value, which is adjusted in a window type comparator electronic circuit to detect any important degrading condition of the capacitance value in a detector-cable system. The ''in-situ'' electronic testing method of neutron detector performance has been verified in a laboratory atmosphere to be a potential method to detect any significant change in the capacitance value of a nuclear sensor and its connecting cable, also checking: detector disconnections, cable disconnections, length changes of the connecting cable, electric short-opened circuits in the sensor channel, and any electrical trouble in the detector-connector-cable system. The experimental practices were carried out by simulation of several electric changes in a nuclear sensor-cable system from a linear D.C. channel which measures reactor power during nuclear reactor operation. It was made at the Training Reactor Electronic Laboratory. The results and conclusions obtained at the Laboratory were proved, satisfactorily, in the Electronic Instrumentation of Budapest Technical University Training Reactor, Hungary

Modified method for combined DNA and RNA isolation from peanut and other oil seeds.

Science.gov (United States)

Dang, Phat M; Chen, Charles Y

2013-02-01

Isolation of good quality RNA and DNA from seeds is difficult due to high levels of polysaccharides, polyphenols, and lipids that can degrade or co-precipitate with nucleic acids. Standard RNA extraction methods utilizing guanidinium-phenol-chloroform extraction has not shown to be successful. RNA isolation from plant seeds is a prerequisite for many seed specific gene expression studies and DNA is necessary in marker-assisted selection and other genetic studies. We describe a modified method to isolate both RNA and DNA from the same seed tissue and have been successful with several oil seeds including peanut, soybean, sunflower, canola, and oil radish. An additional LiCl precipitation step was added to isolate both RNA and DNA from the same seed tissues. High quality nucleic acids were observed based on A(260)/A(280) and A(260)/A(230) ratios above 2.0 and distinct bands on gel-electrophoresis. RNA was shown to be suitable for reverse transcriptase polymerase chain reaction based on actin or 60S ribosomal primer amplification and DNA was shown to have a single band on gel-electrophoresis analysis. This result shows that RNA and DNA isolated using this method can be appropriate for molecular studies in peanut and other oil containing seeds.

Voltammetric, in-situ spectroelectrochemical and in-situ electrocolorimetric characterization of phthalocyanines

Energy Technology Data Exchange (ETDEWEB)

Koca, Atif [Department of Chemical Engineering, Faculty of Engineering, Marmara University, Goeztepe, 34722 Istanbul (Turkey)], E-mail: akoca@eng.marmara.edu.tr; Bayar, Serife; Dincer, Hatice A. [Department of Chemistry, Technical University of Istanbul, Maslak, 34469 Istanbul (Turkey); Gonca, Erguen [Department of Chemistry, Fatih University, TR34500 B.Cekmece, Istanbul (Turkey)

2009-04-01

In this work, electrochemical, and in-situ spectroelectrochemical characterization of the metallophthalocyanines bearing tetra-(1,1-(dicarbethoxy)-2-(2-methylbenzyl))-ethyl 3,10,17,24-tetra chloro groups were performed. Voltammetric and in-situ spectroelectrochemical measurements show that while cobalt phthalocyanine complex gives both metal-based and ring-based redox processes, zinc and copper phthalocyanines show only ring-based reduction and oxidation processes. The redox processes are generally diffusion-controlled, reversible and one-electron transfer processes. Differently lead phthalocyanine demetallized during second oxidation reaction while it was stable during reduction processes. An in-situ electrocolorimetric method, based on the 1931 CIE (Commission Internationale de l'Eclairage) system of colorimetry, has been applied to investigate the color of the electro-generated anionic and cationic forms of the complexes for the first time in this study.

In situ measurement of the efficiency of filtration installations in the nuclear industry by the soda-fluorescein (uranin) aerosol method: AFNOR standard NFX 44.011

International Nuclear Information System (INIS)

Dupoux, J.

1981-01-01

The in situ measurement of the efficiency of filtering installations in the French nuclear industry using the soda-fluorescein (uraniun) aerosol method is presented. A brief description and the interest of the method are discussed. Its use for the location of defects in filter elements and for in situ control of casing with tandem HEPA filter stages is described

An improved method for isolating viruses from asymptomatic carrier fish

Science.gov (United States)

Amend, Donald F.; Pietsch, John P.

1972-01-01

This paper describes a method using elevated levels of penicillin, streptomycin, and nystatin instead of filters to control bacteria and mold contaminants in specimens processed for virus isolation. Filters were shown to significantly reduce the virus concentration. Virus and tissue cultures were not affected by this procedure. In field tests nearly three times more specimens were positive for virus with this method than with the widely used filter technique. Moreover, the cost of materials was less. This method is recommended for inspection and certification purposes.

Evaluation of four methods for estimating leaf area of isolated trees

Science.gov (United States)

P.J. Peper; E.G. McPherson

2003-01-01

The accurate modeling of the physiological and functional processes of urban forests requires information on the leaf area of urban tree species. Several non-destructive, indirect leaf area sampling methods have shown good performance for homogenous canopies. These methods have not been evaluated for use in urban settings where trees are typically isolated and...

A comparison of different methods for in-situ determination of heat losses form district heating pipes

Energy Technology Data Exchange (ETDEWEB)

Boehm, Benny [Technical Univ. of Denmark, Dept. of Energy Engineering (Denmark)

1996-11-01

A comparison of different methods for in-situ determination of heat losses has been carried out on a 273 mm transmission line in Copenhagen. Instrumentation includes temperature sensors, heat flux meters and an infrared camera. The methods differ with regard to time consumption and costs of applying the specific method, demand on accuracy of temperature measurements, sensitivity to computational parameters, e.g. the thermal conductivity of the soil, response to transients in water temperature and the ground, and steady state assumptions in the model used in the interpretation of the measurements. Several of the applied methods work well. (au)

Pattern classification by memristive crossbar circuits using ex situ and in situ training

Science.gov (United States)

Alibart, Fabien; Zamanidoost, Elham; Strukov, Dmitri B.

2013-06-01

Memristors are memory resistors that promise the efficient implementation of synaptic weights in artificial neural networks. Whereas demonstrations of the synaptic operation of memristors already exist, the implementation of even simple networks is more challenging and has yet to be reported. Here we demonstrate pattern classification using a single-layer perceptron network implemented with a memrisitive crossbar circuit and trained using the perceptron learning rule by ex situ and in situ methods. In the first case, synaptic weights, which are realized as conductances of titanium dioxide memristors, are calculated on a precursor software-based network and then imported sequentially into the crossbar circuit. In the second case, training is implemented in situ, so the weights are adjusted in parallel. Both methods work satisfactorily despite significant variations in the switching behaviour of the memristors. These results give hope for the anticipated efficient implementation of artificial neuromorphic networks and pave the way for dense, high-performance information processing systems.

Lactobacillus genus identification isolated from gastrointestinal tract of chickens after bee products application using FISH and RTQ PCR methods

Directory of Open Access Journals (Sweden)

Miroslava Kačániová

2013-05-01

Full Text Available The general objective of this study was to examine the effect of bee products on the lactobacilli colonization of chickens. Bee products were administered to both feed mixtures in various amounts in addition to the control group. First experimental group was with propolis in feed mixture with the addition of 400 mg propolis per 1 kg of compound and second group was with pollen in feed mixture with the addition of 450 mg pollen per 1 kg of compound. In this experiment, quantitative counts of lactobacilli in ceca of 49-day-old chicken (Ross 308 using classical and FISH method were investigated. Counts of lactobacilli on MRS agar were monitored. To check the reliability of traditional methods of cultivation samples were evaluated by fluorescence in situ hybridization (FISH. Lactobacillus cells, isolated from gastrointestinal tract, were detected after hybridization of fluorescently labeled probe with bacterial cells. Counts of CFU of lactobacilli were compared in experimental and control treatments, respectively. The lowest count was detected in the control experimental group. The highest count was detected in the third experimental group where was 450 mg of pollen added to 1 kg of feed mixture. Using Real-time PCR method, we identified the species range of the genera Lactobacillus in the intestinal tract of broiler. Detected species from the genus Lactobacillus were L. crispatus, L. salivarius and L. acidophilus.

Solution (in situ leach) mining of uranium: an overview

International Nuclear Information System (INIS)

Kuhaida, A.J. Jr.; Kelly, M.J.

1978-01-01

Increases in the demand for and price of uranium have made in-situ mining an attractive alternative to the open-pit and underground U mining methods. Up to 50% of the known ore-bearing sandstone in the western U.S. can be mined using the in-situ mining method. In-situ mining also offers a significant environmental advantage. Restoration of the contaminated groundwater is discussed

Impact of enumeration method on diversity of Escherichia coli genotypes isolated from surface water.

Science.gov (United States)

Martin, E C; Gentry, T J

2016-11-01

There are numerous regulatory-approved Escherichia coli enumeration methods, but it is not known whether differences in media composition and incubation conditions impact the diversity of E. coli populations detected by these methods. A study was conducted to determine if three standard water quality assessments, Colilert ® , USEPA Method 1603, (modified mTEC) and USEPA Method 1604 (MI), detect different populations of E. coli. Samples were collected from six watersheds and analysed using the three enumeration approaches followed by E. coli isolation and genotyping. Results indicated that the three methods generally produced similar enumeration data across the sites, although there were some differences on a site-by-site basis. The Colilert ® method consistently generated the least diverse collection of E. coli genotypes as compared to modified mTEC and MI, with those two methods being roughly equal to each other. Although the three media assessed in this study were designed to enumerate E. coli, the differences in the media composition, incubation temperature, and growth platform appear to have a strong selective influence on the populations of E. coli isolated. This study suggests that standardized methods of enumeration and isolation may be warranted if researchers intend to obtain individual E. coli isolates for further characterization. This study characterized the impact of three USEPA-approved Escherichia coli enumeration methods on observed E. coli population diversity in surface water samples. Results indicated that these methods produced similar E. coli enumeration data but were more variable in the diversity of E. coli genotypes observed. Although the three methods enumerate the same species, differences in media composition, growth platform, and incubation temperature likely contribute to the selection of different cultivable populations of E. coli, and thus caution should be used when implementing these methods interchangeably for

Inverse PCR-based method for isolating novel SINEs from genome.

Science.gov (United States)

Han, Yawei; Chen, Liping; Guan, Lihong; He, Shunping

2014-04-01

Short interspersed elements (SINEs) are moderately repetitive DNA sequences in eukaryotic genomes. Although eukaryotic genomes contain numerous SINEs copy, it is very difficult and laborious to isolate and identify them by the reported methods. In this study, the inverse PCR was successfully applied to isolate SINEs from Opsariichthys bidens genome in Eastern Asian Cyprinid. A group of SINEs derived from tRNA(Ala) molecular had been identified, which were named Opsar according to Opsariichthys. SINEs characteristics were exhibited in Opsar, which contained a tRNA(Ala)-derived region at the 5' end, a tRNA-unrelated region, and AT-rich region at the 3' end. The tRNA-derived region of Opsar shared 76 % sequence similarity with tRNA(Ala) gene. This result indicated that Opsar could derive from the inactive or pseudogene of tRNA(Ala). The reliability of method was tested by obtaining C-SINE, Ct-SINE, and M-SINEs from Ctenopharyngodon idellus, Megalobrama amblycephala, and Cyprinus carpio genomes. This method is simpler than the previously reported, which successfully omitted many steps, such as preparation of probes, construction of genomic libraries, and hybridization.

Conductive cotton prepared by polyaniline in situ polymerization using laccase.

Science.gov (United States)

Zhang, Ya; Dong, Aixue; Wang, Qiang; Fan, Xuerong; Cavaco-Paulo, Artur; Zhang, Ying

2014-09-01

The high-redox-potential catalyst laccase, isolated from Aspergillus, was first used as a biocatalyst in the oxidative polymerization of water-soluble conductive polyaniline, and then conductive cotton was prepared by in situ polymerization under the same conditions. The polymerization of aniline was performed in a water dispersion of sodium dodecylbenzenesulfonate (SDBS) micellar solution with atmospheric oxygen serving as the oxidizing agent. This method is ecologically clean and permits a greater degree of control over the kinetics of the reaction. The conditions for polyaniline synthesis were optimized. Characterizations of the conducting polyaniline and cotton were carried out using Fourier transform infrared spectroscopy, UV-vis spectroscopy, cyclic voltammetry, the fabric induction electrostatic tester, and the far-field EMC shielding effectiveness test fixture.

DOE In Situ Remediation Integrated Program

International Nuclear Information System (INIS)

Yow, J.L. Jr.

1993-01-01

The In Situ Remediation Integrated Program (ISRP) supports and manages a balanced portfolio of applied research and development activities in support of DOE environmental restoration and waste management needs. ISRP technologies are being developed in four areas: containment, chemical and physical treatment, in situ bioremediation, and in situ manipulation (including electrokinetics). the focus of containment is to provide mechanisms to stop contaminant migration through the subsurface. In situ bioremediation and chemical and physical treatment both aim to destroy or eliminate contaminants in groundwater and soils. In situ manipulation (ISM) provides mechanisms to access contaminants or introduce treatment agents into the soil, and includes other technologies necessary to support the implementation of ISR methods. Descriptions of each major program area are provided to set the technical context of the ISM subprogram. Typical ISM needs for major areas of in situ remediation research and development are identified

Applications of in situ hybridization to plant-improvement

International Nuclear Information System (INIS)

Abbasi, F.M.

2004-01-01

In situ hybridization is a powerful method for characteristic alien addition and substitution lines. RFLP analysis can identify the presence of a particular individual chromosome, but whether they are as a pair or as a single chromosome cannot be determined. In situ hybridization has become established as an essential method in cell and molecular biology. It is able to link DNA sequences with their organization and physical position. The rate of technology-development in the field of in situ hybridization has been rapid: radioactive probes are now rarely used, while labeling methods, fluorochromes, chromosomes and tissue-preparation methods, microscope and imaging technology have all useful in functional genomics and localization of transgenes on the chromosomes. (author)

Final Report Scalable Analysis Methods and In Situ Infrastructure for Extreme Scale Knowledge Discovery

Energy Technology Data Exchange (ETDEWEB)

O' Leary, Patrick [Kitware, Inc., Clifton Park, NY (United States)

2017-09-13

The primary challenge motivating this project is the widening gap between the ability to compute information and to store it for subsequent analysis. This gap adversely impacts science code teams, who can perform analysis only on a small fraction of the data they calculate, resulting in the substantial likelihood of lost or missed science, when results are computed but not analyzed. Our approach is to perform as much analysis or visualization processing on data while it is still resident in memory, which is known as in situ processing. The idea in situ processing was not new at the time of the start of this effort in 2014, but efforts in that space were largely ad hoc, and there was no concerted effort within the research community that aimed to foster production-quality software tools suitable for use by Department of Energy (DOE) science projects. Our objective was to produce and enable the use of production-quality in situ methods and infrastructure, at scale, on DOE high-performance computing (HPC) facilities, though we expected to have an impact beyond DOE due to the widespread nature of the challenges, which affect virtually all large-scale computational science efforts. To achieve this objective, we engaged in software technology research and development (R&D), in close partnerships with DOE science code teams, to produce software technologies that were shown to run efficiently at scale on DOE HPC platforms.

A novel method for the in situ determination of concentration gradients in the electrolyte of Li-ion Batteries

NARCIS (Netherlands)

Zhou, J.; Danilov, D.; Notten, P.H.L.

2006-01-01

An electrochemical method has been developed for the in situ determination of concentration gradients in the electrolyte of sealed Li-ion batteries by measuring the potential difference between microreference electrodes. Formulas relating the concentration gradient and the potential difference

Localization of mRNA in vertebrate axonal compartments by in situ hybridization.

Science.gov (United States)

Sotelo-Silveira, José Roberto; Calliari, Aldo; Kun, Alejandra; Elizondo, Victoria; Canclini, Lucía; Sotelo, José Roberto

2011-01-01

The conclusive demonstration of RNA in vertebrate axons by in situ hybridization (ISH) has been elusive. We review the most important reasons for difficulties, including low concentration of axonal RNAs, localization in specific cortical domains, and the need to isolate axons. We demonstrate the importance of axon micro-dissection to obtain a whole mount perspective of mRNA distribution in the axonal territory. We describe a protocol to perform fluorescent ISH in isolated axons and guidelines for the preservation of structural and molecular integrity of cortical RNA-containing domains (e.g., Periaxoplasmic Ribosomal Plaques, or PARPs) in isolated axoplasm.

Comparative studies of two methods for miRNA isolation from milk whey.

Science.gov (United States)

Jin, Xiao-lu; Wei, Zi-hai; Liu, Lan; Liu, Hong-yun; Liu, Jian-xin

2015-06-01

MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS(®) followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS(®) followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100).

Comparative studies of two methods for miRNA isolation from milk whey*

Science.gov (United States)

Jin, Xiao-lu; Wei, Zi-hai; Liu, Lan; Liu, Hong-yun; Liu, Jian-xin

2015-01-01

MicroRNAs (miRNAs) from milk whey have been considered for their potential as noninvasive biomarkers for milk quality control and disease diagnosis. However, standard protocols for miRNA isolation and quantification from milk whey are not well established. The objective of this study was to compare two methods for the isolation of miRNAs from milk whey. These two methods were modified phenol-based technique (Trizol LS® followed by phenol precipitation, the TP method) and combined phenol and column-based approach (Trizol LS® followed by cleanup using the miRNeasy kit, the TM method). Yield and quality of RNA were rigorously measured using a NanoDrop ND-1000 spectrophotometer and then the distribution of RNA was precisely detected in a Bioanalyzer 2100 instrument by microchip gel electrophoresis. Several endogenous miRNAs (bta-miR-141, bta-miR-146a, bta-miR-148a, bta-miR-200c, bta-miR-362, and bta-miR-375) and an exogenous spike-in synthetic control miRNA (cel-miR-39) were quantified by real-time polymerase chain reaction (PCR) to examine the apparent recovery efficiency of milk whey miRNAs. Both methods could successfully isolate sufficient small RNA (whey, and their yields were quite similar. However, the quantification results show that the total miRNA recovery efficiency by the TM method is superior to that by the TP method. The TM method performed better than the TP for recovery of milk whey miRNA due to its consistency and good repeatability in endogenous and spike-in miRNA recovery. Additionally, quantitative recovery analysis of a spike-in miRNA may be more accurate to reflect the milk whey miRNA recovery efficiency than using traditional RNA quality analysis instruments (NanoDrop or Bioanalyzer 2100). PMID:26055915

Assignment of Alzheimer's presenilin-2 (PS-2) gene to 1q42.1 by fluorescence in situ hybridization.

Science.gov (United States)

Takano, T; Sahara, N; Yamanouchi, Y; Mori, H

1997-01-17

Presenilin-2 (PS-2) was suggested to be localized on 1q31-42 based on linkage analysis and cDNA cloning. The final identification of PS-2 as the causal gene for early-onset familial Alzheimer's disease in Voga-German pedigrees was concluded based on the point mutation found in the candidate cDNA isolated from this familial AD. We present evidence of its physical genome mapping of PS-2 on chromosome 1q42.1 by fluorescence in situ hybridization method.

Optimum method to determine radioactivity in large tracts of land. In-situ gamma spectroscopy or sampling followed by laboratory measurement

International Nuclear Information System (INIS)

Bronson, Frazier

2008-01-01

In the process of decommissioning contaminated facilities, and in the conduct of normal operations involving radioactive material, it is frequently required to show that large areas of land are not contaminated, or if contaminated that the amount is below an acceptable level. However, it is quite rare for the radioactivity in the soil to be uniformly distributed. Rather it is generally in a few isolated and probably unknown locations. One way to ascertain the status of the land concentration is to take soil samples for subsequent measurement in the laboratory. Another way is to use in-situ gamma spectroscopy. In both cases, the non-uniform distribution of radioactivity can greatly compromise the accuracy of the assay, and makes uncertainty estimates much more complicated than simple propagation of counting statistics. This paper examines the process of determining the best way to estimate the activity on the tract of land, and gives quantitative estimates of measurement uncertainty for various conditions of radioactivity. When the distribution of radioactivity in the soil is not homogeneous, the sampling uncertainty is likely to be larger than the in-situ measurement uncertainty. (author)

A rapid, efficient, and economic device and method for the isolation and purification of mouse islet cells.

Directory of Open Access Journals (Sweden)

Yin Zongyi

Full Text Available Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods. Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P 1000 islets. In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.

Fine typing of methicillin-resistant Staphylococcus aureus isolates using direct repeat unit and staphylococcal interspersed repeat unit typing methods.

Science.gov (United States)

Ho, Cheng-Mao; Ho, Mao-Wang; Li, Chi-Yuan; Lu, Jang-Jih

2015-08-01

Methicillin-resistant Staphylococcus aureus (MRSA) typing is an important epidemiologic tool for monitoring trends and preventing outbreaks. However, the efficiency of various MRSA typing methods for each SCCmec MRSA isolate is rarely evaluated. A total of 157 MRSA isolates from four different regions in Taiwan were typed with five different molecular methods, including SCCmec typing, multilocus sequence typing (MLST), spa typing, mec-associated direct repeat unit (dru) copy number determination, and staphylococcal interspersed repeat unit (SIRU) profiling. There were four SCCmec types, eight MLST types, 15 spa types, 11 dru types, and 31 SIRU profiles. The most common type determined by each molecular typing method was SCCmec III (115 isolates, 73.2%), ST239 (99 isolates, 63.1%), t037 (107 isolates, 68.2%), 14 dru copies (76 isolates, 48.4%), and SIRU profile 3013722 (102 isolates, 65%), respectively. When using the combination of MLST, spa typing, and dru copy number, ST5-t002-4 (n = 8), ST239-t037-14 (n = 68), ST59-t437-9 (n = 9), and ST59-t437-11 (n = 6) were found to be the most common types of SCCmec types II (n = 9), III (n = 115), IV (n = 21), and VT (n = 11) isolates, respectively. SCCmec type III isolates were further classified into 11 dru types. Of the 21 SCCmec type IV isolates, 14 SIRU profiles were found. Seven SIRU patterns were observed in the 11 SCCmec type VT isolates. Different typing methods showed a similar Hunter-Gaston discrimination index among the 157 MRSA isolates. However, dru and SIRU typing methods had a better discriminatory power for SCCmec type III and SCCmec types IV and VT isolates, respectively, suggesting that dru and SIRU can be used to further type these isolates. Copyright © 2013. Published by Elsevier B.V.

Stress and Deformation Analysis in Base Isolation Elements Using the Finite Element Method

Directory of Open Access Journals (Sweden)

Claudiu Iavornic

2011-01-01

Full Text Available In Modern tools as Finite Element Method can be used to study the behavior of elastomeric isolation systems. The simulation results obtained in this way provide a large series of data about the behavior of elastomeric isolation bearings under different types of loads and help in taking right decisions regarding geometrical optimizations needed for improve such kind of devices.

Nuclear waste management by in-situ melting

International Nuclear Information System (INIS)

Angelo, J.A. Jr.

1976-01-01

A systematic assessment of the in-situ melting concept as an ultimate waste disposal option shows that the placement of solidifed, high-level radioactive wastes in an in-situ melting cavity with a crushed rock backfill not only eliminates the major deficiencies inherent in other in-situ melting schemes, but also satisfies reasonable criteria for ultimate disposal. In-situ melting reduces the waste isolation time requirements to several hundred years. Calculated spent fuel and processing waste afterheat values assess the role of actinide and cladding material nuclides in creating the total afterheat and provide quantitative variation with time for these values for contamporary and advanced-design fission reactors. The dominant roles of 134 Cs in thermal spectrum reactor afterheats during the first decade of cooling of the actinide nuclides in all typical waste after-heats following a century or two of cooling are identified. The spatial and temporal behavior of a spherically symmetric waste repository experiencing in-situ melting in an equal density, homogeneous medium for silicate rock and salt is controlled primarily by the overall volumetric thermal source strength, the time-dependent characteristics of the high-level wastes, and the thermophysical properties of the surrounding rock environment. Calculations were verified by experimental data. The hazard index for typical high-level wastes is dominated by the fission product nuclides for the first three centuries of decay. It is then controlled by the actinides, especially americium, which dominates for 10,000 years. With in-situ melting, the hazard index for the re-solidifed rock/waste matrix deepunderground falls below the hazard index of naturally occurring uranium ore bodies within a few hundred years, whether or not the more hazardous actinide nuclides are selectively removed from the wastes prior to storage

Remote vacuum or pressure sealing device and method for critical isolated systems

Science.gov (United States)

Brock, James David [Newport News, VA; Keith, Christopher D [Newport News, VA

2012-07-10

A remote vacuum or pressure sealing apparatus and method for making a radiation tolerant, remotely prepared seal that maintains a vacuum or pressure tight seal throughout a wide temperature range. The remote sealing apparatus includes a fixed threaded sealing surface on an isolated system, a gasket, and an insert consisting of a plug with a protruding sample holder. An insert coupling device, provided for inserting samples within the isolated system, includes a threaded fastener for cooperating with the fixed threaded sealing surface on the isolated system. The insert coupling device includes a locating pin for azimuthal orientation, coupling pins, a tooted coaxial socket wrench, and an insert coupling actuator for actuating the coupling pins. The remote aspect of the sealing apparatus maintains the isolation of the system from the user's environment, safely preserving the user and the system from detrimental effect from each respectively.

Air intake shaft performance tests (Shaft 5): In situ data report (May 1988--July 1995). Waste Isolation Pilot Plant (WIPP) Thermal/Structural Interactions Program

International Nuclear Information System (INIS)

Munson, D.E.; Baird, G.T.; Jones, R.L.

1995-07-01

Data are presented from the Air Intake Shaft Test, an in situ test fielded at the Waste Isolation Pilot Plant (WIPP). The construction of this shaft, well after the initial three access shafts, presented an unusual opportunity to obtain valuable detailed data on the mechanical response of a shaft for application to seal design. These data include selected fielding information, test configuration, instrumentation activities, and comprehensive results from a large number of gages. Construction of the test began in December 1987; gage data in this report cover the period from May 1988 through July 1995, with the bulk of the data obtained after obtaining access in November, 1989 and from the heavily instrumented period after remote gage installation between May, 1990, and October, 1991

Air intake shaft performance tests (Shaft 5): In situ data report (May 1988--July 1995). Waste Isolation Pilot Plant (WIPP) Thermal/Structural Interactions Program

Energy Technology Data Exchange (ETDEWEB)

Munson, D.E. [Sandia National Labs., Albuquerque, NM (United States). Repository Isolation Systems Dept.; Hoag, D.L.; Ball, J.R. [RE/SPEC Inc., Albuquerque, NM (United States); Baird, G.T.; Jones, R.L. [Tech Reps, Inc., Albuquerque, NM (United States)

1995-07-01

Data are presented from the Air Intake Shaft Test, an in situ test fielded at the Waste Isolation Pilot Plant (WIPP). The construction of this shaft, well after the initial three access shafts, presented an unusual opportunity to obtain valuable detailed data on the mechanical response of a shaft for application to seal design. These data include selected fielding information, test configuration, instrumentation activities, and comprehensive results from a large number of gages. Construction of the test began in December 1987; gage data in this report cover the period from May 1988 through July 1995, with the bulk of the data obtained after obtaining access in November, 1989 and from the heavily instrumented period after remote gage installation between May, 1990, and October, 1991.

In-situ probe of the response of the Tile Calorimeter to isolated hadrons

CERN Document Server

Jennens, D; The ATLAS collaboration

2013-01-01

The Tile calorimeter is the hadronic central barrel of the calorimeter system of the ATLAS experiment for the LHC at CERN. It is based on a sampling technique where scintillating tiles are embedded in iron absorber plates. The tiles are grouped together in cells which are disposed in three different layers. The cells from the two innermost layers cover a $\\Delta\\eta \\times \\Delta\\phi $ range of 0.1 $\\times$ 0.1, while the outermost layer covers 0.2 $\\times$ 0.1. An in-situ method to probe the calorimeter response to single charged hadrons can be established by using the ratio of energy measured in the calorimeter cells over the momentum measured by the inner tracking system. This measurement can be used to place constraints on the systematic uncertainty for the jet and tau energy scales. Results from pp collision data from 2010 and 2011 will be shown and discussed as a function of different layer and barrel section. Finally, comparison to MC simulation will prove the good performance of the detector.

Two methods for isolating the lung area of a CT scan for density information

International Nuclear Information System (INIS)

Hedlund, L.W.; Anderson, R.F.; Goulding, P.L.; Beck, J.W.; Effmann, E.L.; Putman, C.E.

1982-01-01

Extracting density information from irregularly shaped tissue areas of CT scans requires automated methods when many scans are involved. We describe two computer methods that automatically isolate the lung area of a CT scan. Each starts from a single, operator specified point in the lung. The first method follows the steep density gradient boundary between lung and adjacent tissues; this tracking method is useful for estimating the overall density and total area of lung in a scan because all pixels within the lung area are available for statistical sampling. The second method finds all contiguous pixels of lung that are within the CT number range of air to water and are not a part of strong density gradient edges; this method is useful for estimating density and area of the lung parenchyma. Structures within the lung area that are surrounded by strong density gradient edges, such as large blood vessels, airways and nodules, are excluded from the lung sample while lung areas with diffuse borders, such as an area of mild or moderate edema, are retained. Both methods were tested on scans from an animal model of pulmonary edema and were found to be effective in isolating normal and diseased lungs. These methods are also suitable for isolating other organ areas of CT scans that are bounded by density gradient edges

Image analyzing method to evaluate in situ bioluminescence from an obligate anaerobe cultivated under various dissolved oxygen concentrations.

Science.gov (United States)

Ninomiya, Kazuaki; Yamada, Ryuji; Matsumoto, Masami; Fukiya, Satoru; Katayama, Takane; Ogino, Chiaki; Shimizu, Nobuaki

2013-02-01

An image analyzing method was developed to evaluate in situ bioluminescence expression, without exposing the culture sample to the ambient oxygen atmosphere. Using this method, we investigated the effect of dissolved oxygen concentration on bioluminescence from an obligate anaerobe Bifidobacterium longum expressing bacterial luciferase which catalyzes an oxygen-requiring bioluminescent reaction. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Thermodynamics parameters of nano-Ni/PS composites prepared by in situ polymerization method

International Nuclear Information System (INIS)

Liao Qilong; Xiong Jie; Ning Haixia

2011-01-01

Spherical nickel nanoparticles with about 75∼200 nm in size were obtained by a liquid reduction method. The nickel nanoparticles/PS composites were synthesized via in situ polymerization method. XRD, FTIR, SEM and TG-DSC were respectively used to measure the properties of nickel nanoparticles, the microstructure of as-prepared composites samples, the distribution of nickel nanoparticles in PS and the thermodynamic parameters of as-prepared composites. The results show that the nickel nanoparticles will enhance the glass transition temperature of nano-Ni/PS composites. The enthalpy of composites is heightened by increasing of doping dose, and it reaches the top when the doping dose is from 1% to 2%. The specific heat of the composites will reduce with the doping dose of nickel nanoparticles increasing. (authors)

Methods of producing alkylated hydrocarbons from an in situ heat treatment process liquid

Science.gov (United States)

Roes, Augustinus Wilhelmus Maria [Houston, TX; Mo, Weijian [Sugar Land, TX; Muylle, Michel Serge Marie [Houston, TX; Mandema, Remco Hugo [Houston, TX; Nair, Vijay [Katy, TX

2009-09-01

A method for producing alkylated hydrocarbons is disclosed. Formation fluid is produced from a subsurface in situ heat treatment process. The formation fluid is separated to produce a liquid stream and a first gas stream. The first gas stream includes olefins. The liquid stream is fractionated to produce at least a second gas stream including hydrocarbons having a carbon number of at least 3. The first gas stream and the second gas stream are introduced into an alkylation unit to produce alkylated hydrocarbons. At least a portion of the olefins in the first gas stream enhance alkylation.

Triplex in-situ hybridization

Science.gov (United States)

Fresco, Jacques R.; Johnson, Marion D.

2002-01-01

Disclosed are methods for detecting in situ the presence of a target sequence in a substantially double-stranded nucleic acid segment, which comprises: a) contacting in situ under conditions suitable for hybridization a substantially double-stranded nucleic acid segment with a detectable third strand, said third strand being capable of hybridizing to at least a portion of the target sequence to form a triple-stranded structure, if said target sequence is present; and b) detecting whether hybridization between the third strand and the target sequence has occured.

NewIn-situ synthesis method of magnesium matrix composites reinforced with TiC particulates

Directory of Open Access Journals (Sweden)

Zhang Xiuqing

2006-12-01

Full Text Available Magnesium matrix composites reinforced with TiC particulates was prepared using a new in-situ synthesis method of remelting and dilution technique. And measurements were performed on the composites. The results of x ray diffraction (XRD analysis confirmed that TiC particulates were synthesized during the sintering process, and they retained in magnesium matrix composites after the remelting and dilution processing. From the microstructure characterization and electron probe microanalysis (EPMA, we could see that fine TiC particulates distributed uniformly in the matrix material.

Transmission electron microscopic method for gene mapping on polytene chromosomes by in situ hybridization

OpenAIRE

Wu, Madeline; Davidson, Norman

1981-01-01

A transmission electron microscope method for gene mapping by in situ hybridization to Drosophila polytene chromosomes has been developed. As electron-opaque labels, we use colloidal gold spheres having a diameter of 25 nm. The spheres are coated with a layer of protein to which Escherichia coli single-stranded DNA is photochemically crosslinked. Poly(dT) tails are added to the 3' OH ends of these DNA strands, and poly(dA) tails are added to the 3' OH ends of a fragmented cloned Drosophila DN...

Neutron reflectivity as method to study in-situ adsorption of phospholipid layers to solid-liquid interfaces

DEFF Research Database (Denmark)

Gutberlet, Thomas; Klösgen, Beate Maria; Krastev, Rumen

2004-01-01

variation. It was observed that the method was capable of visualizing the adsorption of phospholipid layers to different solid-liquid interfaces and to resolve structural details at Angstroem resolution. The results depended strongly on a sufficiently good signal-to-noise ratio of the specific measurements......The use of neutron reflectivity as a method to study in-situ adsorption of phospholipid layers to solid-liquid interfaces was analyzed. The most important advantage of neutron reflectometry is the possibility to very the refractive index of the specific sample by isotope exchange, called contrast...

In situ surface roughness measurement using a laser scattering method

Science.gov (United States)

Tay, C. J.; Wang, S. H.; Quan, C.; Shang, H. M.

2003-03-01

In this paper, the design and development of an optical probe for in situ measurement of surface roughness are discussed. Based on this light scattering principle, the probe which consists of a laser diode, measuring lens and a linear photodiode array, is designed to capture the scattered light from a test surface with a relatively large scattering angle ϕ (=28°). This capability increases the measuring range and enhances repeatability of the results. The coaxial arrangement that incorporates a dual-laser beam and a constant compressed air stream renders the proposed system insensitive to movement or vibration of the test surface as well as surface conditions. Tests were conducted on workpieces which were mounted on a turning machine that operates with different cutting speeds. Test specimens which underwent different machining processes and of different surface finish were also studied. The results obtained demonstrate the feasibility of surface roughness measurement using the proposed method.

Methods for Isolation, Purification, and Propagation of Bacteriophages of Campylobacter jejuni

DEFF Research Database (Denmark)

Gencay, Yilmaz Emre; Birk, Tina; Sørensen, Martine Camilla Holst

2017-01-01

Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-formin...

Radiation-resistant micro-organisms isolated from textiles

International Nuclear Information System (INIS)

Kristensen, H.; Christensen, E.A.

1981-01-01

Towels from private homes and public offices and underwear contaminated by being used by employees at a public health laboratory were examined for occurrence of radiation-resistant bacteria and fungi. Three different methods were used for isolation of the most resistant organisms, one with multiplication of the microbial population prior to an irradiation used for selection, and two witout this multiplication and with the organisms placed on membrane filters or in situ on the textiles, respectively. A total of 44 different strains were isolated. Differences in the three methods used for selection of the most radiation-resistant microorganisms were not reflected in the results. 16 pigmentproducing Gram-positive cocci, tentatively classified as Micrococcus radiodurans, were the most radiation-resistant and were isolated in about half of the examinations. Other Gram-positive cocci, nonspore forming rods, some Nocardia and Candida parapsilosis strains and two Bacillus strains constituted the rest of the collection. With few exceptions dose-response curves for the strains were upward convex. D-6 values determined to be between 1.5 megarad for the most radiation sensitive, a Candida, and 5.7 megarad for the most resistant, tentatively classified as M. radiodurans. The D-6 values for the Bacillus strains were in both cases 1.8 megarad, consistent with a D-value of 0.3 megarad. The same resistance is reported to be the maximum resistance for B. pumilus, strain E601, commonly used as reference strain in the literature on radiation sterilization of medical devices and supplies. (author)

Radiation-resistant micro-organisms isolated from textiles

Energy Technology Data Exchange (ETDEWEB)

Kristensen, H; Christensen, E A [Statens Seruminstitut, Copenhagen (Denmark)

1981-01-01

Towels from private homes and public offices and underwear contaminated by being used by employees at a public health laboratory were examined for occurrence of radiation-resistant bacteria and fungi. Three different methods were used for isolation of the most resistant organisms, one with multiplication of the microbial population prior to an irradiation used for selection, and two witout this multiplication and with the organisms placed on membrane filters or in situ on the textiles, respectively. A total of 44 different strains were isolated. Differences in the three methods used for selection of the most radiation-resistant microorganisms were not reflected in the results. 16 pigmentproducing Gram-positive cocci, tentatively classified as Micrococcus radiodurans, were the most radiation-resistant and were isolated in about half of the examinations. Other Gram-positive cocci, nonspore forming rods, some Nocardia and Candida parapsilosis strains and two Bacillus strains constituted the rest of the collection. With few exceptions dose-response curves for the strains were upward convex. D-6 values determined to be between 1.5 megarad for the most radiation sensitive, a Candida, and 5.7 megarad for the most resistant, tentatively classified as M. radiodurans. The D-6 values for the Bacillus strains were in both cases 1.8 megarad, consistent with a D-value of 0.3 megarad. The same resistance is reported to be the maximum resistance for B. pumilus, strain E601, commonly used as reference strain in the literature on radiation sterilization of medical devices and supplies.

PRODUCTION OF EMBBRYONIC STEM CELLS FROM INNER CELL MASS OF BLASTOCYST ISOLATED BY ENZYMATIC AND IMMUNOSURGERY METHODS

Directory of Open Access Journals (Sweden)

Thomas Mata Hine

2008-03-01

Full Text Available The objective of the research is determining the ICM isolation method to produce ESC. Blastocyst stage of DDy mice embryos were used in this study. Zona pellucida of blastocysts were removed by 0.25% pronase, the ICM isolation were done by enzimatic or immunosurgery method, and then they were cultured in DMEM-high glucose supplemented with mercaptoethanol, gentamycin, fetal bovine serum, and cumulus cells as feeder layer. The result of the research indicated that immunosurgery method yielding attachment rate and number ESC colony 93.85% and 43.08%, respectively, higher (P<0.05 than enzimatic method that weree 79.63% and 18.52%, respectively, but the viability of ICM cells were equal (P >0.05 that are 93.59% in enzymatic method and 98.56% in immunosurgery method. This research concluded that immunosurgery more effective method for isolation of ICM and ESC production than enzymatic method.

Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning

International Nuclear Information System (INIS)

Han, J.H.; Stratowa, C.; Rutter, W.J.

1987-01-01

The authors have cloned a full-length putative rat pancreatic lysophospholipase cDNA by an improved mRNA isolation method and cDNA cloning strategy using [ 32 P]-labelled nucleotides. These new methods allow the construction of a cDNA library from the adult rat pancreas in which the majority of recombinant clones contained complete sequences for the corresponding mRNAs. A previously recognized but unidentified long and relatively rare cDNA clone containing the entire sequence from the cap site at the 5' end to the poly(A) tail at the 3' end of the mRNA was isolated by single-step screening of the library. The size, amino acid composition, and the activity of the protein expressed in heterologous cells strongly suggest this mRNA codes for lysophospholipase

Isolate, functionalize, and release: the ISOFURE platform for the functionalization of nanoparticles

International Nuclear Information System (INIS)

Chirra, Hariharasudhan D.; Spencer, David; Hilt, J. Zach

2012-01-01

Polymer-coated inorganic nanoparticles are widely studied as carrier systems for various biomedical applications. For their translation into clinical applications, it is critical that they remain in a stable, non-agglomerated state and are also produced at a high yield with ease. Herein, we introduce a novel methodology called the isolate-functionalize-release (ISOFURE) strategy to synthesize stable hydrogel-coated nanoparticles in the absence of a stabilizing reagent. As a model platform, we isolated and functionalized gold nanoparticles (GNPs) in a degradable poly(beta amino ester) (PBAE) hydrogel composite. Isolation was done either by adding GNPs to the PBAE hydrogel precursor solution or via in situ precipitation of GNPs inside the pre-synthesized PBAE hydrogel. Atom transfer radical polymerization (ATRP) was used as the model reaction to grow a temperature responsive poly(ethylene glycol) 600 dimethacrylate crosslinked poly(N-isopropyl acrylamide) hydrogel shell over the GNP core. Release of the temperature responsive hydrogel-coated GNPs was done by degrading the PBAE ISOFURE matrix in water. Characterization of the various nanoparticles using UV–Vis spectroscopy confirmed in situ precipitation of GNPs as well as ATRP over the GNPs. Light scattering confirmed the presence of temperature sensitive hydrogel shell over the GNP surface. Aging studies showed that the ISOFURE method based crosslinked hydrogel-coated GNPs showed higher stability and yield than that of the solution-based functionalized GNPs. The use of ISOFURE strategy shows promise as a simple tool for enhanced functionalization and stabilization over various nanocarriers under flexible experimental conditions.

A safe inexpensive method to isolate high quality plant and fungal ...

African Journals Online (AJOL)

The most commonly used plant DNA isolation methods use toxic and hazardous chemicals (phenol, chloroform), which require special equipment to minimize exposure and may limit their use in certain environments. Commercial DNA extraction kits are convenient and usually safe, but their availability to certain developing ...

Comparative miRNA Analysis of Urine Extracellular Vesicles Isolated through Five Different Methods

Directory of Open Access Journals (Sweden)

Felix Royo

2016-12-01

Full Text Available Urine extracellular vesicles are a valuable low-invasive source of information, especially for the cells of the genitourinary tract. In the search for biomarkers, different techniques have been developed to isolate and characterize the cargo of these vesicles. In the present work, we compare five of these different isolation methods (three commercial isolation kits, ultracentrifugation, and lectin-based purification and perform miRNA profiling using a multiplex miRNA assay. The results showed high correlation through all isolation techniques, and 48 out of 68 miRNAs were detected above the detection limit at least 10 times. The results obtained by multiplex assay were validated through Taqman qPCR. In addition, using this technique combined with a clinically friendly extracellular vesicle (uEV-enrichment method, we performed the analysis of selected miRNAs in urine from patients affected with bladder cancer, benign prostate hyperplasia, or prostate cancer. Importantly, we found that those miRNAs could be detected in almost 100% of the samples, and no significant differences were observed between groups. Our results support the feasibility of analyzing exosomes-associated miRNAs using a methodology that requires a small volume of urine and is compatible with a clinical environment and high-throughput analysis.

Early in-situ measurements program for the Waste Isolation Pilot Plant

International Nuclear Information System (INIS)

Wowak, W.E.

1979-06-01

The technical basis and description of measurements for the early in-situ measurements program at the WIPP are described and a proposed organizational structure is presented. Measurements are needed for verification of design predictions and also for a prelude to the main experiment program. The design verification measurements will be concentrated in the first shaft and the underground support and access areas. Early experiments will be concentrated in the test drifts on the storage horizons. Recommendations are made to DOE for appropriate division of responsibility among Bechtel, the technical support contractor, the instrumentation contractor, and Sandia

Evaluation and analysis of geological condition of in-situ fragmentation leaching uranium

International Nuclear Information System (INIS)

Yang Jianming; Tan Kaixuan; Huang Xiaonai

2003-01-01

The ore geological condition, hydrogeological condition, engineering geological condition and technological mineralogical character of in-situ fragmentation leaching uranium are analyzed, and it is considered that the implementation of in-situ fragmentation leaching uranium technology is decided by different geological factor. Previously prospecting and geological condition evaluation of uranium ore is based on traditional mining method. If in-situ fragmentation leaching uranium method is adopted, one must re-evaluate previously prospected deposits before they are mined, or one must evaluate new prospecting deposits according to geological conditions of in-situ fragmentation leaching uranium method. The feasibility evaluation method of uranium deposit by in-situ fragmentation leaching uranium put forward by B. N. Mociniets is introducd, and it is considered that B. N. Mociniets method has guidable significance for geological condition evaluation before uranium deposits are mined. A feasibility study is done by applying B. N. Mociniets method to a uranium deposit. (authors)

Three methods for in situ cross-linking of polyvinyl alcohol films for application as ion-conducting membranes in potassium hydroxide electrolyte. [battery separators

Science.gov (United States)

Philipp, W. H.; Hsu, L. C.

1979-01-01

Three methods of in situ cross-linking polyvinyl alcohol films are presented. They are: (1) acetalization with a dialdehyde such as glutaraldehyde, (2) acetalization with aldehyde groups formed by selective oxidative cleaving of the few percent of 1,2 diol units present in polyvinyl alcohol, and (3) cross-linking by hydrogen abstraction by reaction with hydrogen atoms and hydroxyl radicals from irradiated water. For the third method, improvement in film conductivity in KOH solution at the expense of mechanical strength is obtained by the presence of polyacrylic acid in the polyvinyl alcohol films. Resistivities in 45 percent KOH are given for in situ cross-linked films prepared by each of the three methods.

An improved culture method for selective isolation of Campylobacter jejuni from wastewater

Directory of Open Access Journals (Sweden)

Jinyong Kim

2016-08-01

Full Text Available Campylobacter jejuni is one of the leading foodborne pathogens worldwide. C. jejuni is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently-available culture-based isolation methods are not highly effective for wastewater samples due to the low number of C. jejuni in the midst of competing bacteria. To detect and isolate C. jejuni from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin, to which C. jejuni is intrinsically resistant. The selectivity of each enrichment condition was measured with Ct value using quantitative real-time PCR (qRT-PCR, and multiplex PCR to determine Campylobacter species. In addition, the efficacy of Campylobacter isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of C. jejuni. In particular, rifampicin supplementation and an increased culture temperature (i.e., 42°C had a decisive effect on the selective enrichment of C. jejuni from wastewater. The results of 16S rDNA sequencing also revealed that Enterococcus spp. and Pseudomonas aeruginosa are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate Campylobacter from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious Campylobacter from wastewater.

Instrumentation of the thermal/structural interactions in situ tests at the Waste Isolation Pilot Plant (WIPP)

Energy Technology Data Exchange (ETDEWEB)

Munson, D.E. [Sandia National Labs., Albuquerque, NM (United States). Repository Isolation Systems Div.; Hoag, D.L.; Blankenship, D.A.; DeYonge, W.F.; Schiermeister, D.M. [RE/SPEC, Inc., Albuquerque, NM (United States); Jones, R.L.; Baird, G.T. [Tech Reps, Inc., Albuquerque, NM (United States)

1997-04-01

The Department of Energy has constructed the Waste Isolation Pilot Plant (WIPP) to develop the technology for the disposal of radioactive waste from defense programs. Sandia National Laboratories had the responsibility for the experimental activities at the WIPP and fielded several large-scale Thermal/Structural Interactions (TSI) in situ tests to validate techniques used to predict repository performance. The instrumentation of these tests involved the placement of over 4,200 gages including room closure gages, borehole extensometers, stress gages, borehole inclinometers, fixed reference gages, borehole strain gages, thermocouples, thermal flux meters, heater power gages, environmental gages, and ventilation gages. Most of the gages were remotely read instruments that were monitored by an automated data acquisition system, but manually read instruments were also used to provide early deformation information and to provide a redundancy of measurement for the remote gages. Instruments were selected that could operate in the harsh environment of the test rooms and that could accommodate the ranges of test room responses predicted by pretest calculations. Instruments were tested in the field prior to installation at the WIPP site and were modified to improve their performance. Other modifications were made to gages as the TSI tests progressed using knowledge gained from test maintenance. Quality assurance procedures were developed for all aspects of instrumentation including calibration, installation, and maintenance. The instrumentation performed exceptionally well and has produced a large quantity of quality information.

Instrumentation of the thermal/structural interactions in situ tests at the Waste Isolation Pilot Plant (WIPP)

International Nuclear Information System (INIS)

Munson, D.E.; Jones, R.L.; Baird, G.T.

1997-04-01

The Department of Energy has constructed the Waste Isolation Pilot Plant (WIPP) to develop the technology for the disposal of radioactive waste from defense programs. Sandia National Laboratories had the responsibility for the experimental activities at the WIPP and fielded several large-scale Thermal/Structural Interactions (TSI) in situ tests to validate techniques used to predict repository performance. The instrumentation of these tests involved the placement of over 4,200 gages including room closure gages, borehole extensometers, stress gages, borehole inclinometers, fixed reference gages, borehole strain gages, thermocouples, thermal flux meters, heater power gages, environmental gages, and ventilation gages. Most of the gages were remotely read instruments that were monitored by an automated data acquisition system, but manually read instruments were also used to provide early deformation information and to provide a redundancy of measurement for the remote gages. Instruments were selected that could operate in the harsh environment of the test rooms and that could accommodate the ranges of test room responses predicted by pretest calculations. Instruments were tested in the field prior to installation at the WIPP site and were modified to improve their performance. Other modifications were made to gages as the TSI tests progressed using knowledge gained from test maintenance. Quality assurance procedures were developed for all aspects of instrumentation including calibration, installation, and maintenance. The instrumentation performed exceptionally well and has produced a large quantity of quality information

The influence of the roll diameter in flat rolling of of superconducting in situ and ex situ MgB2 tape

DEFF Research Database (Denmark)

Hancock, Michael Halloway; Bay, Niels

2007-01-01

, 150 and 210 mm in each step. The investigation has shown that the in situ powder is more readily compacted than the ex situ powder, with an average increase of relative density after mechanical processing of 37% for in situ powder and 19% for ex situ powder. Statistical analysis showed that the choice......Applying the powder in tube (PIT) method, single-filament MgB2/Fe wire and tape has been manufactured applying both the ex situ and the in situ approach. The influence of the roll diameter in three-step flat rolling on the powder density and critical temperature has been examined using rolls of 70...... roll in the first and second reductions followed by the 150 mm or 210 mm roll in the last reduction was the optimum strategy for both powder types. AC susceptibility testing showed that for the in situ tapes there was no correlation between the powder density and the critical temperature. For ex situ...

Reduced satellite cell number in situ in muscular contractures from children with cerebral palsy.

Science.gov (United States)

Dayanidhi, Sudarshan; Dykstra, Peter B; Lyubasyuk, Vera; McKay, Bryon R; Chambers, Henry G; Lieber, Richard L

2015-07-01

Satellite cells (SC) are quiescent adult muscle stem cells critical for postnatal development. Children with cerebral palsy have impaired muscular growth and develop contractures. While flow cytometry previously demonstrated a reduced SC population, extracellular matrix abnormalities may influence the cell isolation methods used, systematically isolating fewer cells from CP muscle and creating a biased result. Consequently, the purpose of this study was to use immunohistochemistry on serial muscle sections to quantify SC in situ. Serial cross-sections from human gracilis muscle biopsies (n = 11) were labeled with fluorescent antibodies for Pax7 (SC transcriptional marker), laminin (basal lamina), and 4',6-diamidino-2-phenylindole (nuclei). Fluorescence microscopy under high magnification was used to identify SC based on labeling and location. Mean SC/100 myofibers was reduced by ∼70% (p muscle growth and apparent decreased responsiveness of CP muscle to exercise. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

In-situ calibration of nuclear-plant platinum resistance thermometers using Johnson noise methods. Final report

International Nuclear Information System (INIS)

Blalock, T.V.; Shepard, R.L.

1983-06-01

Methods for in situ calibration of reactor plant platinum resistance thermometers using Johnson noise measurements were tested in the laboratory and in two operating reactors: Diablo Canyon and Sequoyah. The Johnson noise methods provide an absolute measurement of the thermometer temperature and can be compared with the dc calibration of the thermometers to confirm their calibration without removing the thermometers from the plant coolant loops. Inaccuracies of less than 0.1% were obtained with these methods for ideal situations where the noise measuring equipment could be connected directly to the installed thermometer terminals. For measurements made at the ends of long extension cables, inaccuracies were 0.5 to 1.0%, at best. Extension cables must be optimized and well characterized electrically to achieve such accuracies. Other factors that affect the accuracy of these methods were evaluated

An Improved Quick Method for the Isolation of Total RNA from Cotton ...

African Journals Online (AJOL)

David PANG

2011-11-02

Nov 2, 2011 ... in liquid nitrogen in a mortar and pestle and stored until. RNA isolation. ... our laboratory for microarray analysis, cDNA pyro- sequencing studies and construction ..... Economic and rapid method for extracting cotton genomic ...

Identification of MYCN gene amplification in neuroblastoma using chromogenic in situ hybridization (CISH): an alternative and practical method.

Science.gov (United States)

Bhargava, Rohit; Oppenheimer, Orit; Gerald, William; Jhanwar, Suresh C; Chen, Beiyun

2005-06-01

Chromogenic in situ hybridization (CISH) is a recently developed technique, which utilizes the general principles of in situ hybridization and a detection system similar to immunohistochemistry. To assess the utility of CISH for analysis of MYCN gene amplification, we compared this assay with established diagnostic assays such as Southern blot analysis (SB) and fluorescent in situ hybridization (FISH). CISH was performed on 67 cases of neuroblastoma using tissue microarray (65 cases) and whole tissue sections (2 cases). Unequivocal, high-level amplification (> or =10 gene copies per tumor nucleus) was identified in 19 of 67 (28.4%) tumors. Two (3%) tumors showed low-level amplification (6-9 gene copies per tumor nucleus). No amplification was seen in 46 of 67 (68.6%) tumors. SB data were available in 44 tumors. Forty-one of the 44 tumors (93%) showed concordant results between CISH and SB. Three tumors showed MYCN amplification by CISH but no amplification by SB, most likely due to dilution effect of nonneoplastic tissue in the test samples. Two of these three tumors also showed MYCN amplification by FISH, and the third tumor was not analyzed by FISH. FISH data were available in total of 30 tumors. All 30 tumors showed concordant results between CISH and FISH for classifying a tumor as MYCN amplified or not amplified. We conclude that CISH is an accurate method for determining MYCN gene amplification, with added advantages that make it a more practically useful method.

Study on the fabrication of Al matrix composites strengthened by combined in-situ alumina particle and in-situ alloying elements

International Nuclear Information System (INIS)

Huang Zanjun; Yang Bin; Cui Hua; Zhang Jishan

2003-01-01

A new idea to fabricate aluminum matrix composites strengthened by combined in-situ particle strengthening and in-situ alloying has been proposed. Following the concept of in-situ alloying and in-situ particle strengthening, aluminum matrix composites reinforced by Cu and α-Al 2 O 3 particulate (material I) and the same matrix reinforced by Cu, Si alloying elements and α-Al 2 O 3 particulate (material II) have been obtained. SEM observation, EDS and XRD analysis show that the alloy elements Cu and Si exist in the two materials, respectively. In-situ Al 2 O 3 particulates are generally spherical and their mean size is less than 0.5 μm. TEM observation shows that the in-situ α-Al 2 O 3 particulates have a good cohesion with the matrix. The reaction mechanism of the Al 2 O 3 particulate obtained by this method was studied. Thermodynamic considerations are given to the in-situ reactions and the distribution characteristic of in-situ the α-Al 2 O 3 particulate in the process of solidification is also discussed

Comparison of different DNA isolation methods and use of dodecyle trimethyl ammonium bromide (DTAB for the isolation of DNA from meat products

Directory of Open Access Journals (Sweden)

Yusuf OZsENSOY

2016-12-01

Conclusion: DNA isolation kit, another best method, is recommended due to quality and quantity of DNA for researchers who do not want that phenol/chloroform method have toxic substances. This study is also the first study in which DTAB method is used for DNA extraction from meat products. [J Adv Vet Anim Res 2016; 3(4.000: 368-374

Isolation of Native Soil Microorganisms with Potential for Breaking Down Biodegradable Plastic Mulch Films Used in Agriculture

Science.gov (United States)

Bailes, Graham; Lind, Margaret; Ely, Andrew; Powell, Marianne; Moore-Kucera, Jennifer; Miles, Carol; Inglis, Debra; Brodhagen, Marion

2013-01-01

Fungi native to agricultural soils that colonized commercially available biodegradable mulch (BDM) films were isolated and assessed for potential to degrade plastics. Typically, when formulations of plastics are known and a source of the feedstock is available, powdered plastic can be suspended in agar-based media and degradation determined by visualization of clearing zones. However, this approach poorly mimics in situ degradation of BDMs. First, BDMs are not dispersed as small particles throughout the soil matrix. Secondly, BDMs are not sold commercially as pure polymers, but rather as films containing additives (e.g. fillers, plasticizers and dyes) that may affect microbial growth. The procedures described herein were used for isolates acquired from soil-buried mulch films. Fungal isolates acquired from excavated BDMs were tested individually for growth on pieces of new, disinfested BDMs laid atop defined medium containing no carbon source except agar. Isolates that grew on BDMs were further tested in liquid medium where BDMs were the sole added carbon source. After approximately ten weeks, fungal colonization and BDM degradation were assessed by scanning electron microscopy. Isolates were identified via analysis of ribosomal RNA gene sequences. This report describes methods for fungal isolation, but bacteria also were isolated using these methods by substituting media appropriate for bacteria. Our methodology should prove useful for studies investigating breakdown of intact plastic films or products for which plastic feedstocks are either unknown or not available. However our approach does not provide a quantitative method for comparing rates of BDM degradation. PMID:23712218

Isolation of native soil microorganisms with potential for breaking down biodegradable plastic mulch films used in agriculture.

Science.gov (United States)

Bailes, Graham; Lind, Margaret; Ely, Andrew; Powell, Marianne; Moore-Kucera, Jennifer; Miles, Carol; Inglis, Debra; Brodhagen, Marion

2013-05-10

Fungi native to agricultural soils that colonized commercially available biodegradable mulch (BDM) films were isolated and assessed for potential to degrade plastics. Typically, when formulations of plastics are known and a source of the feedstock is available, powdered plastic can be suspended in agar-based media and degradation determined by visualization of clearing zones. However, this approach poorly mimics in situ degradation of BDMs. First, BDMs are not dispersed as small particles throughout the soil matrix. Secondly, BDMs are not sold commercially as pure polymers, but rather as films containing additives (e.g. fillers, plasticizers and dyes) that may affect microbial growth. The procedures described herein were used for isolates acquired from soil-buried mulch films. Fungal isolates acquired from excavated BDMs were tested individually for growth on pieces of new, disinfested BDMs laid atop defined medium containing no carbon source except agar. Isolates that grew on BDMs were further tested in liquid medium where BDMs were the sole added carbon source. After approximately ten weeks, fungal colonization and BDM degradation were assessed by scanning electron microscopy. Isolates were identified via analysis of ribosomal RNA gene sequences. This report describes methods for fungal isolation, but bacteria also were isolated using these methods by substituting media appropriate for bacteria. Our methodology should prove useful for studies investigating breakdown of intact plastic films or products for which plastic feedstocks are either unknown or not available. However our approach does not provide a quantitative method for comparing rates of BDM degradation.

Determination of the ruminant origin of bone particles using fluorescence in situ hybridization (FISH).

Science.gov (United States)

Lecrenier, M C; Ledoux, Q; Berben, G; Fumière, O; Saegerman, C; Baeten, V; Veys, P

2014-07-17

Molecular biology techniques such as PCR constitute powerful tools for the determination of the taxonomic origin of bones. DNA degradation and contamination by exogenous DNA, however, jeopardise bone identification. Despite the vast array of techniques used to decontaminate bone fragments, the isolation and determination of bone DNA content are still problematic. Within the framework of the eradication of transmissible spongiform encephalopathies (including BSE, commonly known as "mad cow disease"), a fluorescence in situ hybridization (FISH) protocol was developed. Results from the described study showed that this method can be applied directly to bones without a demineralisation step and that it allows the identification of bovine and ruminant bones even after severe processing. The results also showed that the method is independent of exogenous contamination and that it is therefore entirely appropriate for this application.

A safe inexpensive method to isolate high quality plant and fungal ...

African Journals Online (AJOL)

STORAGESEVER

2008-08-18

Aug 18, 2008 ... quality DNA from plant and fungal species. This method uses potassium acetate to remove proteins and polysaccharides in an SDS extraction buffer. Further DNA purification is achieved using a low salt. CTAB treatment. This SDS/CTAB protocol was used to isolate high quality genomic DNA subject to.

An in situ method for real-time monitoring of soil gas diffusivity

Science.gov (United States)

Laemmel, Thomas; Maier, Martin; Schack-Kirchner, Helmer; Lang, Friederike

2016-04-01

Soil aeration is an important factor for the biogeochemistry of soils. Generally, gas exchange between soil and atmosphere is assumed to be governed by molecular diffusion and by this way fluxes can be calculated using by Fick's Law. The soil gas diffusion coefficient DS represents the proportional factor between the gas flux and the gas concentration gradient in the soil and reflects the ability of the soil to "transport passively" gas through the soil. One common way to determine DS is taking core samples in the field and measuring DS in the lab. Unfortunately this method is destructive and laborious and it can only reflect a small fraction of the whole soil. As a consequence, uncertainty about the resulting effective diffusivity on the profile scale, i.e. the real aeration status remains. We developed a method to measure and monitor DS in situ. The set-up consists of a custom made gas sampling device, the continuous injection of an inert tracer gas and inverse gas transport modelling in the soil. The gas sampling device has seven sampling depths (from 0 to -43 cm of depth) and can be easily installed into vertical holes drilled by an auger, which allows for fast installation of the system. Helium (He) as inert tracer gas was injected continuously at the lower end of the device. The resulting steady state distribution of He was used to deduce the DS depth distribution of the soil. For Finite Element Modeling of the gas-sampling-device/soil system the program COMSOL was used. We tested our new method both in the lab and in a field study and compared the results with a reference lab method using soil cores. DS profiles obtained by our in-situ method were consistent with DS profiles determined based on soil core analyses. Soil gas profiles could be measured with a temporal resolution of 30 minutes. During the field study, there was an important rain event and we could monitor the decrease in soil gas diffusivity in the top soil due to water infiltration. The effect

Damage and in-situ annealing during ion implantation

International Nuclear Information System (INIS)

Sadana, D.K.; Washburn, J.; Byrne, P.F.; Cheung, N.W.

1982-11-01

Formation of amorphous (α) layers in Si during ion implantation in the energy range 100 keV-11 MeV and temperature range liquid nitrogen (LN)-100 0 C has been investigated. Cross-sectional transmission electron microscopy (XTEM) shows that buried amorphous layers can be created for both room temperature (RT) and LN temperature implants, with a wider 100 percent amorphous region for the LN cooled case. The relative narrowing of the α layer during RT implantation is attributed to in-situ annealing. Implantation to the same fluence at temperatures above 100 0 C does not produce α layers. To further investigate in situ annealing effects, specimens already containing buried α layers were further irradiated with ion beams in the temperature range RT-400 0 C. It was found that isolated small α zones (less than or equal to 50 diameter) embedded in the crystalline matrix near the two α/c interfaces dissolved into the crystal but the thickness of the 100 percent α layer was not appreciably affected by further implantation at 200 0 C. A model for in situ annealing during implantation is presented

A novel In-situ Enzymatic Cleaning Method for Reducing Membrane Fouling in Membrane Bioreactors (MBRs

Directory of Open Access Journals (Sweden)

M. R. Bilad

2016-05-01

Full Text Available A novel in-situ enzymatic cleaning method was developed for fouling control in membrane bioreactors (MBRs. It is achieved by bringing the required enzymes near the membrane surface by pulling the enzymes to a magnetic membrane (MM surface by means of magnetic forces, exactly where the cleaning is required. To achieve this, the enzyme was coupled to a magnetic nanoparticle (MNP and the membrane it self was loaded with MNP. The magnetic activity was turned by means of an external permanent magnet. The effectiveness of concept was tested in a submerged membrane filtration using the model enzyme-substrate of Bacillus subitilis xylanase-arabinoxylan. The MM had almost similar properties compared to the unloaded ones, except for its well distributed MNPs. The enzyme was stable during coupling conditions and the presence of coupling could be detected using a high-performance anion-exchange chromatography (HPAEC analysis and Fourier transform infrared spectroscopy (FTIR. The system facilitated an in-situ enzymatic cleaning and could be effectively applied for control fouling in membrane bioreactors (MBRs.

An intercomparison of three methods for the large-scale isolation of oceanic dissolved organic matter

Science.gov (United States)

Green, Nelson W.; Perdue, E. Michael; Aiken, George R.; Butler, Kenna D.; Chen, Hongmei; Dittmar, Thorsten; Niggemann, Jutta; Stubbins, Aron

2014-01-01

Dissolved organic matter (DOM) was isolated from large volumes of deep (674 m) and surface (21 m) ocean water via reverse osmosis/electrodialysis (RO/ED) and two solid-phase extraction (SPE) methods (XAD-8/4 and PPL) at the Natural Energy Laboratory of Hawaii Authority (NELHA). By applying the three methods to common water samples, the efficiencies of XAD, PPL and RO/ED DOM isolation were compared. XAD recovered 42% of dissolved organic carbon (DOC) from deep water (25% with XAD-8; 17% with XAD-4) and 30% from surface water (16% with XAD-8; 14% with XAD-4). PPL recovered 61 ± 3% of DOC from deep water and 61% from surface water. RO/ED recovered 82 ± 3% of DOC from deep water, 14 ± 3% of which was recovered in a sodium hydroxide rinse, and 75 ± 5% of DOC from surface water, with 12 ± 2% in the sodium hydroxide rinse. The highest recoveries of all were achieved by the sequential isolation of DOC, first with PPL and then via RO/ED. This combined technique recovered 98% of DOC from a deep water sample and 101% of DOC from a surface water sample. In total, 1.9, 10.3 and 1.6 g-C of DOC were collected via XAD, PPL and RO/ED, respectively. Rates of DOC recovery using the XAD, PPL and RO/ED methods were 10, 33 and 10 mg-C h− 1, respectively. Based upon C/N ratios, XAD isolates were heavily C-enriched compared with water column DOM, whereas RO/ED and PPL ➔ RO/ED isolate C/N values were most representative of the original DOM. All techniques are suitable for the isolation of large amounts of DOM with purities suitable for most advanced analytical techniques. Coupling PPL and RO/ED techniques may provide substantial progress in the search for a method to quantitatively isolate oceanic DOC, bringing the entirety of the DOM pool within the marine chemist's analytical window.

Evaluation and optimization of nucleic acid extraction methods for the molecular analysis of bacterial communities associated with corrored steel

NARCIS (Netherlands)

Marty, F.; Ghiglione, J.-F.; Païsse, S.; Gueuné, H.; Quillet, L.; van Loosdrecht, M.C.M.; Muyzer, G.

2012-01-01

Different DNA and RNA extraction approaches were evaluated and protocols optimized on in situ corrosion products from carbon steel in marine environments. Protocols adapted from the PowerSoil DNA/RNA Isolation methods resulted in the best nucleic acid (NA) extraction performances (ie combining high

Characterization of vitrified soil produced by in situ vitrification

International Nuclear Information System (INIS)

Timmerman, C.L.; Lokken, R.O.

1984-01-01

Radioactive or other hazardous wastes buried at waste disposal sites may require further stabilization to secure the isolation of these wastes from the environment. One method of waste stabilization being developed is in situ vitrification. This process involves the in-place melting of buried wastes and the surrounding soil to produce a glass and crystalline waste form. Engineering-scale and pilot-scale demonstrations of this concept with soil contaminated with nonradioactive, hazardous species (Cs, Sr, Ru, Pb, Cd, etc.) were performed. These demonstrations provided information on species migration, crystalline-phase formation, and waste form durability. In addition to the nonradioactive tests, a crucible-scale melt of soil spiked with radioactive uranium, plutonium, and cesium was leach tested. The results show that hazardous waste components are retained in the product. The durability of the waste form in both the vitreous and the crystalline phases is similar to that of Pyrex glass

Characterization of vitrified soil produced by in-situ vitrification

International Nuclear Information System (INIS)

Timmerman, C.L.; Lokken, R.O.

1983-01-01

Radioactive or other hazardous wastes buried at waste-disposal sites may require further stabilization to secure the isolation of these wastes from the environment. One method of waste stabilization being developed is in-situ vitrification. This process involves the in-place melting of buried wastes and the surrounding soil to produce a glass and crystalline waste form. Engineering-scale and pilot-scale demonstrations of this concept with soil contaminated with nonradioactive, hazardous species (Cs, Sr, Ru, Pb, Cd, etc.) were performed. These demonstrations provided information on species migration, crystalline phase formation, and waste form durability. In addition to the nonradioactive tests, a crucible-scale melt of soil spiked with radioactive uranium, plutonium, and cesium was leach tested. The results show that hazardous waste components are retained in the product. The durability of the waste form in both the vitreous and crystalline phases is similar to that of pyrex glass

In-Situ Burning of Crude Oil on Water

DEFF Research Database (Denmark)

van Gelderen, Laurens

in the small scale water basin. Boilovers were also observed during the burning of a heavy crude oil with a substantial light fraction without a water layer, however, which suggests that water is not essential for boilover occurrence. Further studies are required to determine the conditions under which......The fire dynamics and fire chemistry of in-situ burning of crude oil on water was studied in order to improve predictions on the suitability of this oil spill response method. For this purpose, several operational parameters were studied to determine the factors that control the burning efficiency...... of in-situ burning, i.e. the amount of oil (in wt%) removed from the water surface by the burning process. The burning efficiency is the main parameter for expressing the oil removal effectiveness of in-situ burning as response method and is thus relevant for suitability predictions of in-situ burning...

Application of the nuclear x-ray fluorescence method to prospecting for gold in-situ

International Nuclear Information System (INIS)

Zhang, Y.; Xie, T.; Zhou, S.; Ge, L.

1989-01-01

Arsenic and chalcophile elements are often associated with gold, and can be considered indicator elements when prospecting for gold deposits. The nuclear geophysics X-ray fluorescence method can be used to search for hidden gold deposits by measuring fluorescence intensities of the indicator elements in situ. The method can speed geologic investigation and reduce exploration cost. Three types of portable radioisotope X-ray fluorescence analyzers, designed and manufactured by Chengdu College of Geology and Chongqing Geological Instrument Factory, are briefly introduced. These analyzers are widely used in different stages of geologic investigation for gold in China. In the two case histories presented five anomalous zones of X-ray fluorescence intensity related to gold mineralization are located and one hidden gold deposit is discovered with gold content of 23 g/t

In Situ Coupling of Ultrasound to Electro- and Photo-Deposition Methods for Materials Synthesis

Directory of Open Access Journals (Sweden)

Agnieszka Magdziarz

2017-01-01

Full Text Available This short review provides the current state-of-the-art of in situ coupling of ultrasound to chemical deposition methods. A synergetic action of the ultrasound and light radiation or electrical fields may result in new powerful methodologies, and these include sonophotodeposition and sonoelectrodeposition processes. The effect of ultrasound is explained on the base of different physical mechanisms emerging from cavitation phenomenon. Some possible mechanisms of the interactions between ultrasound and photochemical and electrochemical processes are discussed here. The application of sonophotodeposition and sonoelectrodeposition as green energy sources in the syntheses of different nanomaterials is also reviewed.

ADVANCED SEISMIC BASE ISOLATION METHODS FOR MODULAR REACTORS

International Nuclear Information System (INIS)

Blanford, E.; Keldrauk, E.; Laufer, M.; Mieler, M.; Wei, J.; Stojadinovic, B.; Peterson, P.F.

2010-01-01

Advanced technologies for structural design and construction have the potential for major impact not only on nuclear power plant construction time and cost, but also on the design process and on the safety, security and reliability of next generation of nuclear power plants. In future Generation IV (Gen IV) reactors, structural and seismic design should be much more closely integrated with the design of nuclear and industrial safety systems, physical security systems, and international safeguards systems. Overall reliability will be increased, through the use of replaceable and modular equipment, and through design to facilitate on-line monitoring, in-service inspection, maintenance, replacement, and decommissioning. Economics will also receive high design priority, through integrated engineering efforts to optimize building arrangements to minimize building heights and footprints. Finally, the licensing approach will be transformed by becoming increasingly performance based and technology neutral, using best-estimate simulation methods with uncertainty and margin quantification. In this context, two structural engineering technologies, seismic base isolation and modular steel-plate/concrete composite structural walls, are investigated. These technologies have major potential to (1) enable standardized reactor designs to be deployed across a wider range of sites, (2) reduce the impact of uncertainties related to site-specific seismic conditions, and (3) alleviate reactor equipment qualification requirements. For Gen IV reactors the potential for deliberate crashes of large aircraft must also be considered in design. This report concludes that base-isolated structures should be decoupled from the reactor external event exclusion system. As an example, a scoping analysis is performed for a rectangular, decoupled external event shell designed as a grillage. This report also reviews modular construction technology, particularly steel-plate/concrete construction using

ADVANCED SEISMIC BASE ISOLATION METHODS FOR MODULAR REACTORS

Energy Technology Data Exchange (ETDEWEB)

E. Blanford; E. Keldrauk; M. Laufer; M. Mieler; J. Wei; B. Stojadinovic; P.F. Peterson

2010-09-20

Advanced technologies for structural design and construction have the potential for major impact not only on nuclear power plant construction time and cost, but also on the design process and on the safety, security and reliability of next generation of nuclear power plants. In future Generation IV (Gen IV) reactors, structural and seismic design should be much more closely integrated with the design of nuclear and industrial safety systems, physical security systems, and international safeguards systems. Overall reliability will be increased, through the use of replaceable and modular equipment, and through design to facilitate on-line monitoring, in-service inspection, maintenance, replacement, and decommissioning. Economics will also receive high design priority, through integrated engineering efforts to optimize building arrangements to minimize building heights and footprints. Finally, the licensing approach will be transformed by becoming increasingly performance based and technology neutral, using best-estimate simulation methods with uncertainty and margin quantification. In this context, two structural engineering technologies, seismic base isolation and modular steel-plate/concrete composite structural walls, are investigated. These technologies have major potential to (1) enable standardized reactor designs to be deployed across a wider range of sites, (2) reduce the impact of uncertainties related to site-specific seismic conditions, and (3) alleviate reactor equipment qualification requirements. For Gen IV reactors the potential for deliberate crashes of large aircraft must also be considered in design. This report concludes that base-isolated structures should be decoupled from the reactor external event exclusion system. As an example, a scoping analysis is performed for a rectangular, decoupled external event shell designed as a grillage. This report also reviews modular construction technology, particularly steel-plate/concrete construction using

Phenotypic methods for detection of various β-lactamases in Gram-negative clinical isolates: Need of the hour

Directory of Open Access Journals (Sweden)

Neena V Nagdeo

2012-01-01

Full Text Available Background: Many clinical laboratories have problems detecting various β-lactamases. Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions. It is more imperative to use various phenotypic methods for detection of various β-lactamases in routine microbiology laboratory on day-to-day basis to prevent antimicrobial resistance by evidence-based judicious use of antimicrobials. Aims: In view of the multidrug-resistant organisms being reported world over, we planned a cross-sectional prospective analytical study to determine resistance mechanism by various β-lactamases in Gram-negative clinical isolates using various phenotypic methods. Materials and Methods: All nonrepeat, nonenteric clinical isolates of Gram-negative bacilli, resistant to at least two third-generation cephalosporins, were first screened by Novel disc placement method, and isolates showing multiple mechanisms of resistance and reduced zone of inhibition for imipenem were further confirmed for AmpC and metallo β-lactamases. Statistical Analysis: All the data was managed and analyzed in Microsoft Excel. Results: Out of 807 isolates tested, as many as 795 (98.51% revealed the presence of extended-spectrum β-lactamases (ESBLs. Only 10 isolates of Escherichia coli and 2 of Klebsiella pneumoniae did not show production of ESBL. A total of 450 (55.76% isolates produced single enzyme,while 345 (42.75% strains revealed multiple enzyme production simultaneously. Only ESBL production was seen in 315 (39.03% strains, only AmpC in 75 (9.29% and only MBL in 60 (7.44% strains, while ESBL and AmpC together were seen in 219 (27.14% and AmpC plus MBL in 92 (11.40% strains. However, ESBL plus MBL were never observed together. All three enzymes were simultaneously detected in 34 (4.21% strains. Conclusion: This innovative method of disc placement makes it easy, affordable, and reliable method for routine use by basic

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

DEFF Research Database (Denmark)

van Rijk, A.; Svenstroup-Poulsen, T.; Jones, M.

2010-01-01

within the reach of every pathology laboratory. Design and Methods Our study was initiated to determine the consistency between chromogenic in situ hybridization and fluorescence in situ hybridization, both using split-signal probes developed for the detection of chromosomal breaks. Five hundred...... and actual signal were compared to the original fluorescence hybridization results. In addition, hematoxylin background staining intensity and signal intensity of the double-staining chromogenic in situ hybridization procedure were analyzed. Results With respect to the presence or absence of chromosomal...

A high-throughput splinkerette-PCR method for the isolation and sequencing of retroviral insertion sites

DEFF Research Database (Denmark)

Uren, Anthony G; Mikkers, Harald; Kool, Jaap

2009-01-01

sites has been a major limitation to performing screens on this scale. Here we present a method for the high-throughput isolation of insertion sites using a highly efficient splinkerette-PCR method coupled with capillary or 454 sequencing. This protocol includes a description of the procedure for DNA......Insertional mutagens such as viruses and transposons are a useful tool for performing forward genetic screens in mice to discover cancer genes. These screens are most effective when performed using hundreds of mice; however, until recently, the cost-effective isolation and sequencing of insertion...

Hydrodistillation-adsorption method for the isolation of water-soluble, non-soluble and high volatile compounds from plant materials.

Science.gov (United States)

Mastelić, J; Jerković, I; Blazević, I; Radonić, A; Krstulović, L

2008-08-15

Proposed method of hydrodistillation-adsorption (HDA) on activated carbon and hydrodistillation (HD) with solvent trap were compared for the isolation of water-soluble, non-soluble and high volatile compounds, such as acids, monoterpenes, isothiocyanates and others from carob (Certonia siliqua L.), rosemary (Rosmarinus officinalis L.) and rocket (Eruca sativa L.). Isolated volatiles were analyzed by GC and GC/MS. The main advantages of HDA method over ubiquitous HD method were higher yields of volatile compounds and their simultaneous separation in three fractions that enabled more detail analyses. This method is particularly suitable for the isolation and analysis of the plant volatiles with high amounts of water-soluble compounds. In distinction from previously published adsorption of remaining volatile compounds from distillation water on activated carbon, this method offers simultaneous hydrodistillation and adsorption in the same apparatus.

A simple, fast and accurate in-situ method to measure the rate of transport of redox species through membranes for lithium batteries

Science.gov (United States)

Meddings, Nina; Owen, John R.; Garcia-Araez, Nuria

2017-10-01

Lithium ion conducting membranes are important to protect the lithium metal electrode and act as a barrier to crossover species such as polysulphides in Li-S systems, redox mediators in Li-O2 cells or dissolved cathode species or electrolyte oxidation products in high voltage Li-ion batteries. We present an in-situ method for measuring permeability of membranes to crossover redox species. The method employs a 'Swagelok' cell design equipped with a glassy carbon working electrode, in which redox species are placed initially in the counter electrode compartment only. Permeability through the membrane, which separates working and counter electrodes, is determined using a square wave voltammetry technique that allows the concentration of crossover redox species to be evaluated over time with very high precision. We test the method using a model and well-behaved electrochemical system to demonstrate its sensitivity, reproducibility and reliability relative to alternative approaches. This new method offers advantages in terms of small electrolyte volume, and simple, fast, quantitative and in-situ measurement.

In Situ TEM Electrical Measurements

DEFF Research Database (Denmark)

Canepa, Silvia; Alam, Sardar Bilal; Ngo, Duc-The

2016-01-01

understanding of complex physical and chemical interactions in the pursuit to optimize nanostructure function and device performance. Recent developments of sample holder technology for TEM have enabled a new field of research in the study of functional nanomaterials and devices via electrical stimulation...... influence the sample by external stimuli, e.g. through electrical connections, the TEM becomes a powerful laboratory for performing quantitative real time in situ experiments. Such TEM setups enable the characterization of nanostructures and nanodevices under working conditions, thereby providing a deeper...... and measurement of the specimen. Recognizing the benefits of electrical measurements for in situ TEM, many research groups have focused their effort in this field and some of these methods have transferred to ETEM. This chapter will describe recent advances in the in situ TEM investigation of nanostructured...

In situ synthesis of protein arrays.

Science.gov (United States)

He, Mingyue; Stoevesandt, Oda; Taussig, Michael J

2008-02-01

In situ or on-chip protein array methods use cell free expression systems to produce proteins directly onto an immobilising surface from co-distributed or pre-arrayed DNA or RNA, enabling protein arrays to be created on demand. These methods address three issues in protein array technology: (i) efficient protein expression and availability, (ii) functional protein immobilisation and purification in a single step and (iii) protein on-chip stability over time. By simultaneously expressing and immobilising many proteins in parallel on the chip surface, the laborious and often costly processes of DNA cloning, expression and separate protein purification are avoided. Recently employed methods reviewed are PISA (protein in situ array) and NAPPA (nucleic acid programmable protein array) from DNA and puromycin-mediated immobilisation from mRNA.

In-situ Raman spectroscopy. A method to study and control the growth of microcrystalline silicon for thin-film solar cells

Energy Technology Data Exchange (ETDEWEB)

Muthmann, Stefan

2012-08-22

This work deals with the design and application of a novel experiment, which enables in-situ Raman measurements during the parallel plate plasma enhanced chemical vapor deposition (PECVD) of {mu}cSi:H. Measurements of the crystalline volume fraction (I{sub C}{sup RS}) and the temperature of a growing film are carried out using the novel setup. To enable in-situ Raman measurement of central regions of the coated substrate in a PECVD system, optical access under normal incidence is necessary. An experimental setup in which an optical feed-through was integrated into a PECVD electrode was developed. This setup introduces a disturbance to the electrical field which sustains the plasma. By designing metallic shields the impact of the feed through was reduced considerably at low optical losses. The homogeneity of films deposited with the novel setup in different growth regimes was studied. A correlation between the magnitude of the inhomogeneity caused by the feed-through and the characteristics of the deposition regimes is found. Raman spectroscopy demands the illumination of a sample with a laser and the collection of the scattered radiation. Due to absorption of the laser light the temperature of the illuminated film is increased. Since the temperature determines the properties of a growing film the laser-induced temperature increase was studied. By pulsing the laser radiation of minimal temperature increase at maximal signal intensity was obtained. The crystalline volume fraction of a growing {mu}cSi:H layer was determined in-situ with the novel setup. A minimal temporal resolution of less than 17.5 s at sufficient signal-to-noise-ratio was achieved, which corresponds to less than 9 nm of deposited material during one measurement interval at the industrial standard growth rate of 0.5 nm/s. The obtained results were compared to depth resolved measurements which were carried out after the deposition. An excellent agreement between both methods validates the reliability

A 3-D aerodynamic method for the analysis of isolated horizontal-axis wind turbines

Energy Technology Data Exchange (ETDEWEB)

Ammara, I.; Masson, C.; Paraschivoiu, I. [Ecole Polytechnique, Montreal (Canada)

1997-12-31

In most existing performance-analysis methods, wind turbines are considered isolated so that interference effects caused by other rotors or by the site topography are neglected. The main objective of this paper is to propose a practical 3-D method suitable for the study of these effects, in order to optimize the arrangement and the positioning of Horizontal-Axis Wind Turbines (HAWTs) in a wind farm. In the proposed methodology, the flow field around isolated HAWTs is predicted by solving the 3-D, time-averaged, steady-state, incompressible, Navier-Stokes equations in which the turbines are represented by distributions of momentum sources. The resulting governing equations are solved using a Control-Volume Finite Element Method (CVFEM). The fundamental aspects related to the development of a practical 3-D method are discussed in this paper, with an emphasis on some of the challenges that arose during its implementation. The current implementation is limited to the analysis of isolated HAWTs. Preliminary results have indicated that, the proposed 3-D method reaches the same level of accuracy, in terms of performance predictions, that the previously developed 2-D axisymmetric model and the well-known momentum-strip theory, while still using reasonable computers resources. It can be considered as a useful tool for the design of HAWTs. Its main advantages, however, are its intrinsic capacity to predict the details of the flow in the wake, and its capabilities of modelling arbitrary wind-turbine arrangements and including ground effects.

Base Isolation for Seismic Retrofitting of a Multiple Building Structure: Evaluation of Equivalent Linearization Method

Directory of Open Access Journals (Sweden)

Massimiliano Ferraioli

2016-01-01

Full Text Available Although the most commonly used isolation systems exhibit nonlinear inelastic behaviour, the equivalent linear elastic analysis is commonly used in the design and assessment of seismic-isolated structures. The paper investigates if the linear elastic model is suitable for the analysis of a seismically isolated multiple building structure. To this aim, its computed responses were compared with those calculated by nonlinear dynamic analysis. A common base isolation plane connects the isolation bearings supporting the adjacent structures. In this situation, the conventional equivalent linear elastic analysis may have some problems of accuracy because this method is calibrated on single base-isolated structures. Moreover, the torsional characteristics of the combined system are significantly different from those of separate isolated buildings. A number of numerical simulations and parametric studies under earthquake excitations were performed. The accuracy of the dynamic response obtained by the equivalent linear elastic model was calculated by the magnitude of the error with respect to the corresponding response considering the nonlinear behaviour of the isolation system. The maximum displacements at the isolation level, the maximum interstorey drifts, and the peak absolute acceleration were selected as the most important response measures. The influence of mass eccentricity, torsion, and high-modes effects was finally investigated.

Waste Isolation Pilot Plant Materials Interface Interactions Test: Papers presented at the Commission of European Communities workshop on in situ testing of radioactive waste forms and engineered barriers

International Nuclear Information System (INIS)

Molecke, M.A.; Sorensen, N.R.

1993-08-01

The three papers in this report were presented at the second international workshop to feature the Waste Isolation Pilot Plant (WIPP) Materials Interface Interactions Test (MIIT). This Workshop on In Situ Tests on Radioactive Waste Forms and Engineered Barriers was held in Corsendonk, Belgium, on October 13--16, 1992, and was sponsored by the Commission of the European Communities (CEC). The Studiecentrum voor Kernenergie/Centre D'Energie Nucleaire (SCK/CEN, Belgium), and the US Department of Energy (via Savannah River) also cosponsored this workshop. Workshop participants from Belgium, France, Germany, Sweden, and the United States gathered to discuss the status, results and overviews of the MIIT program. Nine of the twenty-five total workshop papers were presented on the status and results from the WIPP MIIT program after the five-year in situ conclusion of the program. The total number of published MIIT papers is now up to almost forty. Posttest laboratory analyses are still in progress at multiple participating laboratories. The first MIIT paper in this document, by Wicks and Molecke, provides an overview of the entire test program and focuses on the waste form samples. The second paper, by Molecke and Wicks, concentrates on technical details and repository relevant observations on the in situ conduct, sampling, and termination operations of the MIIT. The third paper, by Sorensen and Molecke, presents and summarizes the available laboratory, posttest corrosion data and results for all of the candidate waste container or overpack metal specimens included in the MIIT program

Ex-situ and in-situ mineral carbonation as a means to sequester carbon dioxide

Energy Technology Data Exchange (ETDEWEB)

Gerdemann, Stephen J.; Dahlin, David C.; O' Connor, William K.; Penner, Larry R.; Rush, G.E.

2004-01-01

The U. S. Department of Energy's Albany Research Center is investigating mineral carbonation as a method of sequestering CO2 from coal-fired-power plants. Magnesium-silicate minerals such as serpentine [Mg3Si2O5(OH)4] and olivine (Mg2SiO4) react with CO2 to produce magnesite (MgCO3), and the calcium-silicate mineral, wollastonite (CaSiO3), reacts to form calcite (CaCO3). It is possible to carry out these reactions either ex situ (above ground in a traditional chemical processing plant) or in situ (storage underground and subsequent reaction with the host rock to trap CO2 as carbonate minerals). For ex situ mineral carbonation to be economically attractive, the reaction must proceed quickly to near completion. The reaction rate is accelerated by raising the activity of CO2 in solution, heat (but not too much), reducing the particle size, high-intensity grinding to disrupt the crystal structure, and, in the case of serpentine, heat-treatment to remove the chemically bound water. All of these carry energy/economic penalties. An economic study illustrates the impact of mineral availability and process parameters on the cost of ex situ carbon sequestration. In situ carbonation offers economic advantages over ex situ processes, because no chemical plant is required. Knowledge gained from the ex situ work was applied to long-term experiments designed to simulate in situ CO2 storage conditions. The Columbia River Basalt Group (CRBG), a multi-layered basaltic lava formation, has potentially favorable mineralogy (up to 25% combined concentration of Ca, Fe2+, and Mg cations) for storage of CO2. However, more information about the interaction of CO2 with aquifers and the host rock is needed. Core samples from the CRBG, as well as samples of olivine, serpentine, and sandstone, were reacted in an autoclave for up to 2000 hours at elevated temperatures and pressures. Changes in core porosity, secondary mineralizations, and both solution and solid chemistry were measured.

Evaluation methods for assessing effectiveness of in situ remediation of soil and sediment contaminated with organic pollutants and heavy metals.

Science.gov (United States)

Song, Biao; Zeng, Guangming; Gong, Jilai; Liang, Jie; Xu, Piao; Liu, Zhifeng; Zhang, Yi; Zhang, Chen; Cheng, Min; Liu, Yang; Ye, Shujing; Yi, Huan; Ren, Xiaoya

2017-08-01

Soil and sediment contamination has become a critical issue worldwide due to its great harm to the ecological environment and public health. In recent years, many remediation technologies including physical, chemical, biological, and combined methods have been proposed and adopted for the purpose of solving the problems of soil and sediment contamination. However, current research on evaluation methods for assessing these remediation technologies is scattered and lacks valid and integrated evaluation methods for assessing the remediation effectiveness. This paper provides a comprehensive review with an environmental perspective on the evaluation methods for assessing the effectiveness of in situ remediation of soil and sediment contaminated with organic pollutants and heavy metals. The review systematically summarizes recent exploration and attempts of the remediation effectiveness assessment based on the content of pollutants, soil and sediment characteristics, and ecological risks. Moreover, limitations and future research needs of the practical assessment are discussed. These limitations are not conducive to the implementation of the abatement and control programs for soil and sediment contamination. Therefore, more attention should be paid to the evaluation methods for assessing the remediation effectiveness while developing new in situ remediation technologies in future research. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of an in situ magnetic beads based RT-PCR method for electrochemiluminescent detection of rotavirus

Science.gov (United States)

Zhan, Fangfang; Zhou, Xiaoming

2012-12-01

Rotaviruses are double-stranded RNA viruses belonging to the family of enteric pathogens. It is a major cause of diarrhoeal disease in infants and young children worldwide. Consequently, rapid and accurate detection of rotaviruses is of great importance in controlling and preventing food- and waterborne diseases and outbreaks. Reverse transcription-polymerase chain reaction (RT-PCR) is a reliable method that possesses high specificity and sensitivity. It has been widely used to detection of viruses. Electrochemiluminescence (ECL) can be considered as an important and powerful tool in analytical and clinical application with high sensitivity, excellent specificity, and low cost. Here we have developed a method for the detection of rotavirus by combining in situ magnetic beads (MBs) based RT-PCR with ECL. RT of rotavirus RNA was carried out in a traditional way and the resulting cDNA was directly amplified on MBs. Forward primers were covalently bounded to MBs and reverse primers were labeled with tris-(2, 2'-bipyridyl) ruthenium (TBR). During the PCR cycling, the TBR labeled products were directly loaded and enriched on the surface of MBs. Then the MBs-TBR complexes could be analyzed by a magnetic ECL platform without any post-modification or post-incubation which avoid some laborious manual operations and achieve rapid yet sensitive detection. In this study, rotavirus from fecal specimens was successfully detected within 2 h, and the limit of detection was estimated to be 104copies/μL. This novel in situ MBs based RT-PCR with ECL detection method can be used for pathogen detection in food safety field and clinical diagnosis.

In situ detection, isolation, and physiological properties of a thin filamentous microorganism abundant in methanogenic granular sludges: a novel isolate affiliated with a clone cluster, the green non-sulfur bacteria, subdivision I.

Science.gov (United States)

Sekiguchi, Y; Takahashi, H; Kamagata, Y; Ohashi, A; Harada, H

2001-12-01

We previously showed that very thin filamentous bacteria affiliated with the division green non-sulfur bacteria were abundant in the outermost layer of thermophilic methanogenic sludge granules fed with sucrose and several low-molecular-weight fatty acids (Y. Sekiguchi, Y. Kamagata, K. Nakamura, A. Ohashi, H. Harada, Appl. Environ. Microbiol. 65:1280-1288, 1999). Further 16S ribosomal DNA (rDNA) cloning-based analysis revealed that the microbes were classified within a unique clade, green non-sulfur bacteria (GNSB) subdivision I, which contains a number of 16S rDNA clone sequences from various environmental samples but no cultured representatives. To investigate their function in the community and physiological traits, we attempted to isolate the yet-to-be-cultured microbes from the original granular sludge. The first attempt at isolation from the granules was, however, not successful. In the other thermophilic reactor that had been treating fried soybean curd-manufacturing wastewater, we found filamentous microorganisms to outgrow, resulting in the formation of projection-like structures on the surface of granules, making the granules look like sea urchins. 16S rDNA-cloning analysis combined with fluorescent in situ hybridization revealed that the projections were comprised of the uncultured filamentous cells affiliated with the GNSB subdivision I and Methanothermobacter-like cells and the very ends of the projections were comprised solely of the filamentous cells. By using the tip of the projection as the inoculum for primary enrichment, a thermophilic, strictly anaerobic, filamentous bacterium, designated strain UNI-1, was successfully isolated with a medium supplemented with sucrose and yeast extract. The strain was a very slow growing bacterium which is capable of utilizing only a limited range of carbohydrates in the presence of yeast extract and produced hydrogen from these substrates. The growth was found to be significantly stimulated when the strain was

Sensitive microculture method for isolation of human immunodeficiency virus type 1 from blood leukocytes.

Science.gov (United States)

Erice, A; Sannerud, K J; Leske, V L; Aeppli, D; Balfour, H H

1992-02-01

A study was conducted to compare our standard culture with a new microculture procedure for isolation of human immunodeficiency virus type 1 (HIV-1) from blood leukocytes. A total of 137 blood specimens from 102 HIV-1 antibody-positive individuals (52 were asymptomatic, 31 were symptomatic, and 19 had AIDS) were cultured in a microculture system in which 10(6) of the patients' peripheral blood mononuclear cells (PBMC) were cocultured with 10(6) phytohemagglutinin (PHA)-stimulated PBMC from an HIV-1 antibody-negative blood donor in 1.2 ml of culture medium. Results were compared with those of a historical control group of 139 standard HIV-1 cultures from 108 HIV-1 antibody-positive subjects (58 were asymptomatic, 36 were symptomatic, and 14 had AIDS). For standard cultures, 10 x 10(6) of the patients' PBMC were cocultured with 5 x 10(6) PHA-stimulated PBMC from an HIV-1 antibody-negative blood donor in 15 ml of culture medium. HIV-1 was isolated in 128 (93%) microcultures and 133 (96%) standard cultures. Both methods identified more than 75% of the positive cultures within 7 days and 100% of the positive cultures within 14 days. The isolation rates for HIV-1 in microcultures compared with standard cultures were 91 versus 93% (specimens from asymptomatic individuals), 93 versus 96% (specimens from symptomatic individuals), and 97 versus 100% (specimens from patients with AIDS). The median time to positivity for both culture methods was 7 days, and this correlated significantly with symptoms and CD4+ cell counts. The microculture method is a sensitive and less expensive system for isolation of HIV-1 from PBMC of HIV-1 antibody-positive individuals, and we recommend it as the culture method of choice, especially for children and patients with AIDS and severe anemia or leukopenia whose blood volume is an important consideration.

A whole-mount in situ hybridization method for microRNA detection in Caenorhabditis elegans.

Science.gov (United States)

Andachi, Yoshiki; Kohara, Yuji

2016-07-01

Whole-mount in situ hybridization (WISH) is an outstanding method to decipher the spatiotemporal expression patterns of microRNAs (miRNAs) and provides important clues for elucidating their functions. The first WISH method for miRNA detection was developed in zebrafish. Although this method was quickly adapted for other vertebrates and fruit flies, WISH analysis has not been successfully used to detect miRNAs in Caenorhabditis elegans Here, we show a novel WISH method for miRNA detection in C. elegans Using this method, mir-1 miRNA was detected in the body-wall muscle where the expression and roles of mir-1 miRNA have been previously elucidated. Application of the method to let-7 family miRNAs, let-7, mir-48, mir-84, and mir-241, revealed their distinct but partially overlapping expression patterns, indicating that miRNAs sharing a short common sequence were distinguishably detected. In pash-1 mutants that were depleted of mature miRNAs, signals of mir-48 miRNA were greatly reduced, suggesting that mature miRNAs were detected by the method. These results demonstrate the validity of WISH to detect mature miRNAs in C. elegans. © 2016 Andachi and Kohara; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Regulation of in situ to invasive breast carcinoma transition

Energy Technology Data Exchange (ETDEWEB)

Polyak, Kornelia; Hu, Min; Yao, Jun; Carroll, Danielle K.; Weremowicz, Stanislawa; Chen, Haiyan; Carrasco, Daniel; Richardson, Andrea; Violette, Shelia; Gelman, Rebecca S.; Bissell, Mina J.; Schnitt, Stuart; Polyak, Kornelia

2008-05-07

The transition of ductal carcinoma in situ (DCIS) to invasive carcinoma is a key event in breast tumor progression that is poorly understood. Comparative molecular analysis of tumor epithelial cells from in situ and invasive tumors has failed to identify consistent tumor stage-specific differences. However, the myoepithelial cell layer, present only in DCIS, is a key distinguishing and diagnostic feature. To determine the contribution of non-epithelial cells to tumor progression, we analyzed the role of myoepithelial cells and fibroblasts in the progression of in situ carcinomas using a xenograft model of human DCIS. Progression to invasion was promoted by fibroblasts, but inhibited by normal myoepithelial cells. The invasive tumor cells from these progressed lesions formed DCIS rather than invasive cancers when re-injected into naive mice. Molecular profiles of myoepithelial and epithelial cells isolated from primary normal and cancerous human breast tissue samples corroborated findings obtained in the xenograft model. These results provide the proof of principle that breast tumor progression could occur in the absence of additional genetic alterations and that tumor growth and progression could be controlled by replacement of normal myoepithelial inhibitory signals.

Regulation of In Situ to Invasive Breast CarcinomaTransition

Energy Technology Data Exchange (ETDEWEB)

Hu, Min; Carroll, Danielle K.; Weremowicz, Stanislawa; Chen,Haiyan; Carrasco, Daniel; Richardson, Andrea; Bissell, Mina; Violette,Shelia; Gelman, Rebecca S.; Schnitt, Stuart; Polyak, Kornelia

2007-03-13

The transition of ductal carcinoma in situ (DCIS) to invasive carcinoma is a key event in breast tumor progression that is poorly understood. Comparative molecular analysis of tumor epithelial cells from in situ and invasive tumors has failed to identify consistent tumor stage-specific differences. However, the myoepithelial cell layer, present only in DCIS, is a key distinguishing and diagnostic feature. To determine the contribution of non-epithelial cells to tumor progression, we analyzed the role of myoepithelial cells and fibroblasts in the progression of in situ carcinomas using a xenograft model of human DCIS. Progression to invasion was promoted by fibroblasts, but inhibited by normal myoepithelial cells. The invasive tumor cells from these progressed lesions formed DCIS rather than invasive cancers when re-injected into naive mice. Molecular profiles of myoepithelial and epithelial cells isolated from primary normal and cancerous human breast tissue samples corroborated findings obtained in the xenograft model. These results provide the proof of principle that breast tumor progression could occur in the absence of additional genetic alterations and that tumor growth and progression could be controlled by replacement of normal myoepithelial inhibitory signals.

Application of in situ current normalized PIGE method for determination of total boron and its isotopic composition

International Nuclear Information System (INIS)

Chhillar, Sumit; Acharya, R.; Sodaye, S.; Pujari, P.K.

2014-01-01

A particle induced gamma-ray emission (PIGE) method using proton beam has been standardized for determination of isotopic composition of natural boron and enriched boron samples. Target pellets of boron standard and samples were prepared in cellulose matrix. The prompt gamma rays of 429 keV, 718 keV and 2125 keV were measured from 10 B(p,αγ) 7 Be, 10 B(p, p'γ) 10 B and 11 B(p, p'γ) 11 B nuclear reactions, respectively. For normalizing the beam current variations in situ current normalization method was used. Validation of method was carried out using synthetic samples of boron carbide, borax, borazine and lithium metaborate in cellulose matrix. (author)

[Comparison of manual and automated (MagNA Pure) nucleic acid isolation methods in molecular diagnosis of HIV infections].

Science.gov (United States)

Alp, Alpaslan; Us, Dürdal; Hasçelik, Gülşen

2004-01-01

Rapid quantitative molecular methods are very important for the diagnosis of human immunodeficiency virus (HIV) infections, assessment of prognosis and follow up. The purpose of this study was to compare and evaluate the performances of conventional manual extraction method and automated MagNA Pure system, for the nucleic acid isolation step which is the first and most important step in molecular diagnosis of HIV infections. Plasma samples of 35 patients in which anti-HIV antibodies were found as positive by microparticule enzyme immunoassay and confirmed by immunoblotting method, were included in the study. The nucleic acids obtained simultaneously by manual isolation kit (Cobas Amplicor, HIV-1 Monitor Test, version 1.5, Roche Diagnostics) and automated system (MagNA Pure LC Total Nucleic Acid Isolation Kit, Roche Diagnostics), were amplified and detected in Cobas Amplicor (Roche Diagnostics) instrument. Twenty three of 35 samples (65.7%) were found to be positive, and 9 (25.7%) were negative by both of the methods. The agreement between the methods were detected as 91.4%, for qualitative results. Viral RNA copies detected by manual and MagNA Pure isolation methods were found between 76.0-7.590.000 (mean: 487.143) and 113.0-20.300.0000 (mean: 2.174.097) copies/ml, respectively. When both of the overall and individual results were evaluated, the number of RNA copies obtained with automatized system, were found higher than the manual method (p<0.05). Three samples which had low numbers of nucleic acids (113, 773, 857, respectively) with MagNA Pure, yielded negative results with manual method. In conclusion, the automatized MagNA Pure system was found to be a reliable, rapid and practical method for the isolation of HIV-RNA.

In-situ load-deformation characterization of the CSM/OCRD jointed test block

International Nuclear Information System (INIS)

Richardson, A.M.; Hustrulid, W.; Brown, S.; Ubbes, W.

1985-01-01

An extensive ambient-temperature test series has recently been completed on a block of Precambrian Gneiss at the Colorado School of Mines (CSM) Experimental Mine in Idaho Springs, Colorado. Block tests came into existence out of a desire to test a relatively large volume of rock and thereby minimize the scaling problems encountered when laboratory test results are used to obtain modeling parameters for full-size structures. A typical block test involves isolation of a large, approximately two-meter cube of rock by cutting slots on four sides and inserting flatjacks for loading. Much interest has recently centered around block tests as a promising method for in-situ characterization of rock-masses for licensing future commercial nuclear-waste repositories in crystalline rock. To date only a few block tests have been conducted

Comparison of the Histopaque-1119 method with the Plasmagel method for separation of blood leukocytes for cytomegalovirus isolation.

OpenAIRE

Slifkin, M; Cumbie, R

1992-01-01

Histopaque-1119 (Sigma Chemical Co., St. Louis, Mo.) and Plasmagel (Cellular Products, Inc., Buffalo, N.Y.) were compared as density gradient separation reagents for the separation of polymorphonuclear leukocytes and mononuclear cells from blood from the isolation of cytomegalovirus (CMV). Of 200 peripheral blood specimens examined, CMV was recovered from 51 by both methods. The time of detection of immunofluorescent sites or a cytopathic effect associated with CMV was similar by each method....

Effects of different processing methods of flaxseed on ruminal degradability and in vitro post-ruminal nutrient disappearance

DEFF Research Database (Denmark)

Lashkari, Saman

2015-01-01

The aim of the study was to determine the effects of different heat-processing methods of flaxseed on the in situ effective dry matter degradability (EDMD) and the in situ effective crude protein degradability (ECPD). The treatments included roasting, steep roasting, rolled roasting, rolled steep...... roasting, microwave irradiation and extrusion. Three rumen-fistulated sheep were used for in situ incubations. Furthermore, the effects of heat-processing methods on post-ruminal in vitro nutrient disappearance and total tract disappearance were measured by a three-step in vitro technique. The seeds were...... roasted and extruded at 140°C to 145°C. One lot of roasted seeds was gradually cooled for about 1 h (roasting) and another lot was held in temperature isolated barrels for 45 min (steep roasting). Moreover, roasted and steep roasted flaxseed was rolled in a roller mill. The lowest and highest EDMD...

Improved Simplified Methods for Effective Seismic Analysis and Design of Isolated and Damped Bridges in Western and Eastern North America

Science.gov (United States)

Koval, Viacheslav

The seismic design provisions of the CSA-S6 Canadian Highway Bridge Design Code and the AASHTO LRFD Seismic Bridge Design Specifications have been developed primarily based on historical earthquake events that have occurred along the west coast of North America. For the design of seismic isolation systems, these codes include simplified analysis and design methods. The appropriateness and range of application of these methods are investigated through extensive parametric nonlinear time history analyses in this thesis. It was found that there is a need to adjust existing design guidelines to better capture the expected nonlinear response of isolated bridges. For isolated bridges located in eastern North America, new damping coefficients are proposed. The applicability limits of the code-based simplified methods have been redefined to ensure that the modified method will lead to conservative results and that a wider range of seismically isolated bridges can be covered by this method. The possibility of further improving current simplified code methods was also examined. By transforming the quantity of allocated energy into a displacement contribution, an idealized analytical solution is proposed as a new simplified design method. This method realistically reflects the effects of ground-motion and system design parameters, including the effects of a drifted oscillation center. The proposed method is therefore more appropriate than current existing simplified methods and can be applicable to isolation systems exhibiting a wider range of properties. A multi-level-hazard performance matrix has been adopted by different seismic provisions worldwide and will be incorporated into the new edition of the Canadian CSA-S6-14 Bridge Design code. However, the combined effect and optimal use of isolation and supplemental damping devices in bridges have not been fully exploited yet to achieve enhanced performance under different levels of seismic hazard. A novel Dual-Level Seismic

[Means and methods of personal hygiene in the experiment with 520-day isolation].

Science.gov (United States)

Shumilina, G A; Shumilina, I V; Solov'eva, S O

2013-01-01

Six volunteers (3 Russians, a Frenchman, an Italian and a Chinese) participated in assessment of the input of sanitation and housekeeping provisions to their wellbeing during 520-day isolation and confinement. Subject of the study was quality and sufficiency of housekeeping agents and procedures as well as more than 60 names of personal hygiene items. The sanitation and housekeeping monitoring involved the clinical, hygienic and microbiological methods, and also consideration of crew comments on the items at their disposal and recommended procedures. Based on the analysis of the functional condition of the integument and oral cavity and entries in the questionnaires, i.e. objective data and subjective feelings, all test subjects remained in the invariably good state. Owing to the application of the selected hygienic means and methods the microbial status of the crew was stable throughout 520-day isolation.

Immediate assessment of in situ gas content using underground manometric desorbometer

Energy Technology Data Exchange (ETDEWEB)

Lunaezewski, L.W.; Mahoney, M.R. [Lunagas Pty. Ltd., Newcastle, NSW (Australia)

1995-12-31

An underground assessment of gas content (within 30 minutes) can be used as a complementary method for the immediate establishment of the magnitudes of in situ gas content at a distance of 5 to 6 metres ahead of a heading face. The indirect method of in situ gas content measurement, using a manometric desorbometer, can be used to establish the direct relationship between underground desorbometer readings and corresponding in situ gas content values for a selected coal seam. 3 refs., 3 figs.

C-STrap Sample Preparation Method--In-Situ Cysteinyl Peptide Capture for Bottom-Up Proteomics Analysis in the STrap Format.

Directory of Open Access Journals (Sweden)

Alexandre Zougman

Full Text Available Recently we introduced the concept of Suspension Trapping (STrap for bottom-up proteomics sample processing that is based upon SDS-mediated protein extraction, swift detergent removal and rapid reactor-type protein digestion in a quartz depth filter trap. As the depth filter surface is made of silica, it is readily modifiable with various functional groups using the silane coupling chemistries. Thus, during the digest, peptides possessing specific features could be targeted for enrichment by the functionalized depth filter material while non-targeted peptides could be collected as an unbound distinct fraction after the digest. In the example presented here the quartz depth filter surface is functionalized with the pyridyldithiol group therefore enabling reversible in-situ capture of the cysteine-containing peptides generated during the STrap-based digest. The described C-STrap method retains all advantages of the original STrap methodology and provides robust foundation for the conception of the targeted in-situ peptide fractionation in the STrap format for bottom-up proteomics. The presented data support the method's use in qualitative and semi-quantitative proteomics experiments.

C-STrap Sample Preparation Method--In-Situ Cysteinyl Peptide Capture for Bottom-Up Proteomics Analysis in the STrap Format.

Science.gov (United States)

Zougman, Alexandre; Banks, Rosamonde E

2015-01-01

Recently we introduced the concept of Suspension Trapping (STrap) for bottom-up proteomics sample processing that is based upon SDS-mediated protein extraction, swift detergent removal and rapid reactor-type protein digestion in a quartz depth filter trap. As the depth filter surface is made of silica, it is readily modifiable with various functional groups using the silane coupling chemistries. Thus, during the digest, peptides possessing specific features could be targeted for enrichment by the functionalized depth filter material while non-targeted peptides could be collected as an unbound distinct fraction after the digest. In the example presented here the quartz depth filter surface is functionalized with the pyridyldithiol group therefore enabling reversible in-situ capture of the cysteine-containing peptides generated during the STrap-based digest. The described C-STrap method retains all advantages of the original STrap methodology and provides robust foundation for the conception of the targeted in-situ peptide fractionation in the STrap format for bottom-up proteomics. The presented data support the method's use in qualitative and semi-quantitative proteomics experiments.

The study of redox mechanism of dobutamine at different pH media by electrochemical and in situ spectroelectrochemical methods

International Nuclear Information System (INIS)

Yang Gongjun; Xu Jingjuan; Chen Hongyuan

2004-01-01

Based on the comprehensive analyses of the experimental results of the electrochemical methods, in situ UV-Vis absorption spectra, in situ electron spin resonance (ESR), and attenuated total-internal reflection (ATR) as well as the calculation of UV-Vis absorption data by PM3 Semi-Empirical method, a reaction mechanism for the redox processes of dobutamine was presented. When the anodic sweep is carried out, dobutamine firstly undergoes a free radical reaction with one-electron and one-proton to form semi-quinone free radicals, which will continuously convert to its corresponding quinone form by further electrochemical oxidation reaction. The formed quinone cannot only undergo a cyclization process by chemical reaction to produce a new compound, which can be reduced at more negative potential, but also be reduced to form dobutamine again when subsequent cathodic sweep is followed. The cyclization rate is depended upon pH values, and it increases with the increase of pH. In neutral medium, the corresponding oxidation form of the cyclization reaction product is easy to convert to melanin

Deep sequencing as a method of typing bluetongue virus isolates.

Science.gov (United States)

Rao, Pavuluri Panduranga; Reddy, Yella Narasimha; Ganesh, Kapila; Nair, Shreeja G; Niranjan, Vidya; Hegde, Nagendra R

2013-11-01

Bluetongue (BT) is an economically important endemic disease of livestock in tropics and subtropics. In addition, its recent spread to temperate regions like North America and Northern Europe is of serious concern. Rapid serotyping and characterization of BT virus (BTV) is an essential step in the identification of origin of the virus and for controlling the disease. Serotyping of BTV is typically performed by serum neutralization, and of late by nucleotide sequencing. This report describes the near complete genome sequencing and typing of two isolates of BTV using Illumina next generation sequencing platform. Two of the BTV RNAs were multiplexed with ten other unknown samples. Viral RNA was isolated and fragmented, reverse transcribed, the cDNA ends were repaired and ligated with a multiplex oligo. The genome library was amplified using primers complementary to the ligated oligo and subjected to single and paired end sequencing. The raw reads were assembled using a de novo method and reference-based assembly was performed based on the contig data. Near complete sequences of all segments of BTV were obtained with more than 20× coverage, and single read sequencing method was sufficient to identify the genotype and serotype of the virus. The two viruses used in this study were typed as BTV-1 and BTV-9E. Copyright © 2013 Elsevier B.V. All rights reserved.

In-situ polymerization PLOT columns I: divinylbenzene

Science.gov (United States)

Shen, T. C.

1992-01-01

A novel method for preparation of porous-layer open-tubular (PLOT) columns is described. The method involves a simple and reproducible, straight-forward in-situ polymerization of monomer directly on the metal tube.

iPBS: a universal method for DNA fingerprinting and retrotransposon isolation.

Science.gov (United States)

Kalendar, Ruslan; Antonius, Kristiina; Smýkal, Petr; Schulman, Alan H

2010-11-01

Molecular markers are essential in plant and animal breeding and biodiversity applications, in human forensics, and for map-based cloning of genes. The long terminal repeat (LTR) retrotransposons are well suited as molecular markers. As dispersed and ubiquitous transposable elements, their "copy and paste" life cycle of replicative transposition leads to new genome insertions without excision of the original element. Both the overall structure of retrotransposons and the domains responsible for the various phases of their replication are highly conserved in all eukaryotes. Nevertheless, up to a year has been required to develop a retrotransposon marker system in a new species, involving cloning and sequencing steps as well as the development of custom primers. Here, we describe a novel PCR-based method useful both as a marker system in its own right and for the rapid isolation of retrotransposon termini and full-length elements, making it ideal for "orphan crops" and other species with underdeveloped marker systems. The method, iPBS amplification, is based on the virtually universal presence of a tRNA complement as a reverse transcriptase primer binding site (PBS) in LTR retrotransposons. The method differs from earlier retrotransposon isolation methods because it is applicable not only to endogenous retroviruses and retroviruses, but also to both Gypsy and Copia LTR retrotransposons, as well as to non-autonomous LARD and TRIM elements, throughout the plant kingdom and to animals. Furthermore, the inter-PBS amplification technique as such has proved to be a powerful DNA fingerprinting technology without the need for prior sequence knowledge.

Simulation of salt behavior using in situ response

International Nuclear Information System (INIS)

Li, W.T.

1986-01-01

The time-dependent nonlinear structural behavior in a salt formation around the openings can be obtained by either performing computational analysis of measuring in situ responses. However, analysis using laboratory test data may often deviate from the actual in situ conditions and geomechanical instruments can provide information only up to the time when the measurements were taken. A method has been suggested for simulating the salt behavior by utilizing the steady-state portion of in situ response history. Governing equations for computational analysis were normalized to the creep constant, the equations were solved, and the analytical response history was then computed in terms of normalized time. By synchronizing the response history obtained from the analysis to the one measured at the site, the creep constant was determined. Then the structural response of the salt was computed. This paper presents an improved method for simulating the salt behavior. In this method, the governing equations are normalized to the creep function, which represents the transient and the steady-state creep behavior. Both the transient and the steady-state portions of in situ response history are used in determining the creep function. Finally, a nonlinear mapping process relating the normalized and real time domains determines the behavior of the salt

A new method for the determination of radionuclide distribution in the soil by in situ gamma-ray spectrometry

International Nuclear Information System (INIS)

Zombori, P.; Andrasi, A.; Nemeth, I.

1992-06-01

A method was searched for to estimate the penetration characteristics of fallout radioactivity, using only spectral information obtained by in situ spectrometric measurements, and avoiding the need for long and tiresome sampling and sample analysis procedures. To speed up the analysis for depth distribution determination of fallout radioactivity in soil, an instrumental method based on the shape of spectra was developed. The ratio of peak to valley (the region between the photopeak and Compton edge) depends on the penetration of radionuclides in soil, providing an estimation of depth profile. These ratios were calculated and the method was tested by actual measurements. (R.P.) 7 refs.; 14 figs.; 2 tabs

Development of an in situ polymeric hydrogel implant of ...

African Journals Online (AJOL)

Purpose: To prepare and characterize in situ gel-forming implants of methylprednisolone for the treatment of spinal cord injuries. Methods: In situ hydrogels of methylprednisolone were prepared by dispersing polylactide glycolic acid (PLGA) polymer and methylprednisolone in N-methyl-pyrrolidone solvent, and subsequent ...

Effect of drying methods on the structure, thermo and functional properties of fenugreek (Trigonella foenum graecum) protein isolate.

Science.gov (United States)

Feyzi, Samira; Varidi, Mehdi; Zare, Fatemeh; Varidi, Mohammad Javad

2018-03-01

Different drying methods due to protein denaturation could alter the functional properties of proteins, as well as their structure. So, this study focused on the effect of different drying methods on amino acid content, thermo and functional properties, and protein structure of fenugreek protein isolate. Freeze and spray drying methods resulted in comparable protein solubility, dynamic surface and interfacial tensions, foaming and emulsifying properties except for emulsion stability. Vacuum oven drying promoted emulsion stability, surface hydrophobicity and viscosity of fenugreek protein isolate at the expanse of its protein solubility. Vacuum oven process caused a higher level of Maillard reaction followed by the spray drying process, which was confirmed by the lower amount of lysine content and less lightness, also more browning intensity. ΔH of fenugreek protein isolates was higher than soy protein isolate, which confirmed the presence of more ordered structures. Also, the bands which are attributed to the α-helix structures in the FTIR spectrum were in the shorter wave number region for freeze and spray dried fenugreek protein isolates that show more possibility of such structures. This research suggests that any drying method must be conducted in its gentle state in order to sustain native structure of proteins and promote their functionalities. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Breeding of in-situ Petroleum Degrading Bacteria in Hangzhou Bay and evaluating for the In-situ repair effect

Science.gov (United States)

Lan, Ru; Lin, Hai; Qiao, Bing; Dong, Yingbo; Zhang, Wei; Chang, Wen

2018-02-01

In this paper, the restoration behaviour of the in-situ microorganisms in seawater and sediments to the marine accident oil spill was researched. The experimental study on the breeding of in-situ petroleum-degrading bacteria in the seawater and sediments of Hangzhou Bay and the restoration of oil spill were carried out. Making use of the reinforced microbial flora, combined with physical and chemical methods in field environment, petroleum degrading and restoration experiment were performed, the effect of the breeding of in-situ degrading bacteria was evaluated, and the standard process of in-situ bacteria sampling, laboratory screening, domestication and degradation efficiency testing were formed. This study laid a foundation for further evaluation of the advantages and disadvantages for the petroleum-degrading bacteria of Hangzhou Bay during the process of in-situ restoration. The results showed that in-situ microbes of Hangzhou Bay could reach the growth peak in 5 days with the suitable environmental factors and sufficient nutrient elements, and the degradation efficiency could reach 65.2% (or 74.8% after acclimation). And also the microbes could adapt to the local sea water and environmental conditions, with a certain degree of degradation. The research results could provide parameter support for causal judgment and quantitative assessment of oil spill damage.

Soil Segregation Methods for Reducing Transportation and Disposal Costs - 13544

Energy Technology Data Exchange (ETDEWEB)

Frothingham, David; Andrews, Shawn; Barker, Michelle; Boyle, James; Buechi, Stephen; Graham, Marc; Houston, Linda; Polek, Michael; Simmington, Robert; Spector, Harold [U.S. Army Corps of Engineers, Buffalo District, 1776 Niagara St., Buffalo, NY 14207 (United States); Elliott, Robert ' Dan' [U.S. Army Reserve, 812A Franklin St.,Worcester, MA 01604 (United States); Durham, Lisa [Argonne National Laboratory, Environmental Science Division, 9700 S. Cass Ave., Argonne, IL 60439 (United States)

2013-07-01

At Formerly Utilized Sites Remedial Action Program (FUSRAP) sites where the selected alternative for contaminated soil is excavation and off-site disposal, the most significant budget items of the remedial action are the costs for transportation and disposal of soil at an off-site facility. At these sites, the objective is to excavate and dispose of only those soils that exceed derived concentration guideline levels. In situ soil segregation using gross gamma detectors to guide the excavation is often challenging at sites where the soil contamination is overlain by clean soil or where the contaminated soil is located in isolated, subsurface pockets. In addition, data gaps are often identified during the alternative evaluation and selection process, resulting in increased uncertainty in the extent of subsurface contamination. In response, the U.S. Army Corps of Engineers, Buffalo District is implementing ex situ soil segregation methods. At the remediated Painesville Site, soils were excavated and fed through a conveyor-belt system, which automatically segregated them into above- and below-cleanup criteria discharge piles utilizing gamma spectroscopy. At the Linde Site and the Shallow Land Disposal Area (SLDA) Site, which are both in the remediation phase, soils are initially segregated during the excavation process using gross gamma detectors and then transported to a pad for confirmatory manual surveying and sampling. At the Linde Site, the ex situ soils are analyzed on the basis of a site-specific method, to establish compliance with beneficial reuse criteria that were developed for the Linde remediation. At the SLDA Site, the ex situ soils are surveyed and sampled based on Multi-Agency Radiation Survey and Site Investigation Manual (MARSSIM) final status survey guidance to demonstrate compliance with the derived concentration guideline levels. At all three sites, the ex situ soils that meet the site- specific DCGLs are retained on-site and used as backfill

Soil Segregation Methods for Reducing Transportation and Disposal Costs - 13544

International Nuclear Information System (INIS)

Frothingham, David; Andrews, Shawn; Barker, Michelle; Boyle, James; Buechi, Stephen; Graham, Marc; Houston, Linda; Polek, Michael; Simmington, Robert; Spector, Harold; Elliott, Robert 'Dan'; Durham, Lisa

2013-01-01

At Formerly Utilized Sites Remedial Action Program (FUSRAP) sites where the selected alternative for contaminated soil is excavation and off-site disposal, the most significant budget items of the remedial action are the costs for transportation and disposal of soil at an off-site facility. At these sites, the objective is to excavate and dispose of only those soils that exceed derived concentration guideline levels. In situ soil segregation using gross gamma detectors to guide the excavation is often challenging at sites where the soil contamination is overlain by clean soil or where the contaminated soil is located in isolated, subsurface pockets. In addition, data gaps are often identified during the alternative evaluation and selection process, resulting in increased uncertainty in the extent of subsurface contamination. In response, the U.S. Army Corps of Engineers, Buffalo District is implementing ex situ soil segregation methods. At the remediated Painesville Site, soils were excavated and fed through a conveyor-belt system, which automatically segregated them into above- and below-cleanup criteria discharge piles utilizing gamma spectroscopy. At the Linde Site and the Shallow Land Disposal Area (SLDA) Site, which are both in the remediation phase, soils are initially segregated during the excavation process using gross gamma detectors and then transported to a pad for confirmatory manual surveying and sampling. At the Linde Site, the ex situ soils are analyzed on the basis of a site-specific method, to establish compliance with beneficial reuse criteria that were developed for the Linde remediation. At the SLDA Site, the ex situ soils are surveyed and sampled based on Multi-Agency Radiation Survey and Site Investigation Manual (MARSSIM) final status survey guidance to demonstrate compliance with the derived concentration guideline levels. At all three sites, the ex situ soils that meet the site- specific DCGLs are retained on-site and used as backfill

Investigation of colistin sensitivity via three different methods in Acinetobacter baumannii isolates with multiple antibiotic resistance.

Science.gov (United States)

Sinirtaş, Melda; Akalin, Halis; Gedikoğlu, Suna

2009-09-01

In recent years there has been an increase in life-threatening infections caused by Acinetobacter baumannii with multiple antibiotic resistance, which has lead to the use of polymyxins, especially colistin, being reconsidered. The aim of this study was to investigate the colistin sensitivity of A. baumannii isolates with multiple antibiotic resistance via different methods, and to evaluate the disk diffusion method for colistin against multi-resistant Acinetobacter isolates, in comparison to the E-test and Phoenix system. The study was carried out on 100 strains of A. baumannii (colonization or infection) isolated from the microbiological samples of different patients followed in the clinics and intensive care units of Uludağ University Medical School between the years 2004 and 2005. Strains were identified and characterized for their antibiotic sensitivity by Phoenix system (Becton Dickinson, Sparks, MD, USA). In all studied A. baumannii strains, susceptibility to colistin was determined to be 100% with the disk diffusion, E-test, and broth microdilution methods. Results of the E-test and broth microdilution method, which are accepted as reference methods, were found to be 100% consistent with the results of the disk diffusion tests; no very major or major error was identified upon comparison of the tests. The sensitivity and the positive predictive value of the disk diffusion method were found to be 100%. Colistin resistance in A. baumannii was not detected in our region, and disk diffusion method results are in accordance with those of E-test and broth microdilution methods.

Membrane solid-phase extraction: Field application for isolation of polycyclic aromatic hydrocarbons from water samples

International Nuclear Information System (INIS)

Furlong, E.T.; Koleis, J.C.; Gates, P.M.

1995-01-01

Solid-phase extraction (SPE) membranes (M-SPE) were used to isolate microgram-per-liter to nanogram-per-liter quantities of polycyclic aromatic hydrocarbons (PAH) in 4- to 8-liter ground-water samples from a crude-oil-contaminated ground-water site near Bemidji, Minnesota. The M-SPE method was evaluated (1) under laboratory conditions using reagent water fortified with individual PAH at 1.23 micrograms per liter, and (2) at the Bemidji site. At the site, ground-water samples were processed and PAH isolated using a M-SPE system connected directly to the well pump. Following sample isolation, all M-SPE samples were extracted using dichloromethane and analyzed by gas chromatography-mass spectrometry with selected-ion monitoring. Operationally, the M-SPE method provided a simple means to isolate PAH on site at the wellhead, particularly for anoxic water samples. Acceptable recoveries, ranging from 56 to over 100 percent, were observed for lower molecular weight PAH (naphthalene to pyrene) using the M-SPE method. Recoveries using M-SPE were somewhat lower, but reproducible, for higher molecular weight PAH (chrysene to benzo[ghi]perylene), ranging from 18 to 56 percent. M-SPE provides the capability to collect and field isolate PAH from a sufficiently large number of samples to identify environmental chemical processes occurring at individual compound concentrations of 50 to 1,200 nanograms per liter. Using M-SPE, the potential for facilitated transport of PAH by in situ-derived dissolved organic carbon (DOC) was evaluated at the site. Plots comparing DOC and PAH concentrations indicate that PAH concentrations increase exponentially with linear increases in DOC concentrations

Development of the integrated in situ Lasagna process

International Nuclear Information System (INIS)

Ho, S.; Athmer, C.; Sheridan, P.

1995-01-01

Contamination in deep, low permeability soils poses a significant technical challenge to in-situ remediation efforts. Poor accessibility to the contaminants and difficulty in uniform delivery of treatment reagents have rendered existing in-situ methods such as bioremediation, vapor extraction, and pump and treat rather ineffective when applied to low permeability soils present at many contaminated sites

Enrichment Method for the Isolation of Bioactive Actinomycetes From Mangrove Sediments of Andaman Islands, India

Directory of Open Access Journals (Sweden)

Baskaran, R.

2011-01-01

Full Text Available Various pre-treatment methods and three different media were employed for the isolation of bioactive actinomycetes from mangrove sediments of Andaman and Nicobar Islands, India. Sediments from four different sites of mangrove forest were collected and pre-treated by dry heat method, and the media were supplemented with cycloheximide 80 µg/mL and nalidixic acid 75 µg/mL. The mean actinomycetes population density in sediment samples were recorded as 22 CFU-10^-6/gm in KUA medium followed by 12 CFU-10^-6/gm in AIA medium and 8 CFU-10^-6/gm in SCA medium. A total of 42 actinomycetes were isolated, and all the isolates were evaluated for their antibacterial activity against pathogenic bacteria on two different media. Among 42 isolates tested, 22 species were found to be antibacterial metabolite producer against test bacteria namely, Staphylococcus aureus, Bacillus subtilis, Salmonella typhi and Klebsiella pneumoniae. Particularly, the actinomycete strains such as A101, A102, A107, A116, A121, A125, A130, F101, F102, F104, F106, De101 and De102 significantly inhibited the growth of all bacteria which were tested. Of these strains, A107 was identified as Streptomyces spp. This strain had the maximum activity against all used pathogens on both medium. Hence, the isolation, characterization and studies of secondary metabolites of actinomycetes from mangrove sediments in Andaman and Nicobar Island could be a pathway for discovery of antibiotics from marine actinomycetes.

Isolation of naphthalene-degrading bacteria from tropical marine sediments

International Nuclear Information System (INIS)

Zhuang, W.-Q.; Tay, J.-H.; Maszenan, A.M.; Tay, S.T.-L.

2003-01-01

Oil pollution is a major environmental concern in many countries, and this has led to a concerted effort in studying the feasibility of using oil-degrading bacteria for bioremediation. Although many oil-degrading bacteria have been isolated from different environments, environmental conditions can impose a selection pressure on the types of bacteria that can reside in a particular environment. This study reports the successful isolation of two indigenous naphthalene-degrading bacteria from oil-contaminated tropical marine sediments by enrichment culture. Strains MN-005 and MN-006 were characterized using an extensive range of biochemical tests. The 16S ribosomal deoxyribonucleic acid (rDNA) sequence analysis was also performed for the two strains. Their naphthalene degradation capabilities were determined using gas chromatography and DAPI counting of bacterial cells. Strains MN-005 and MN-006 are phenotypically and phylogenetically different from each other, and belong to the genera Staphylococcus and Micrococcus, respectively. Strains MN-005 and MN-006 has maximal specific growth rates (μ max ) of 0.082±0.008 and 0.30±0.02 per hour, respectively, and half-saturation constants (K s ) of 0.79±0.10 and 2.52±0.32 mg per litre, respectively. These physiological and growth studies are useful in assessing the potential of these indigenous isolates for in situ or ex situ naphthalene pollutant bioremediation in tropical marine environments. (author)

Cell-cell interactions of isolated and cultured oligodendrocytes: formation of linear occluding junctions and expression of peculiar intramembrane particles.

Science.gov (United States)

Massa, P T; Szuchet, S; Mugnaini, E

1984-12-01

Oligodendrocytes were isolated from lamb brain. Freshly isolated cells and cultured cells, either 1- to 4-day-old unattached or 1- to 5-week-old attached, were examined by thin section and freeze-fracture electron microscopy. Freeze-fracture of freshly isolated oligodendrocytes showed globular and elongated intramembrane particles similar to those previously described in oligodendrocytes in situ. Enrichment of these particles was seen at sites of inter-oligodendrocyte contact. Numerous gap junctions and scattered linear tight junctional arrays were apparent. Gap junctions were connected to blebs of astrocytic plasma membrane sheared off during isolation, whereas tight junctions were facing extracellular space or blebs of oligodendrocytic plasma membrane. Thin sections of cultured, unattached oligodendrocytes showed rounded cell bodies touching one another at points without forming specialized cell junctions. Cells plated on polylysine-coated aclar dishes attached, emanated numerous, pleomorphic processes, and expressed galactocerebroside and myelin basic protein, characteristic markers for oligodendrocytes. Thin sections showed typical oligodendrocyte ultrastructure but also intermediate filaments not present in unattached cultures. Freeze-fracture showed intramembrane particles similar to but more numerous, and with a different fracture face repartition, than those seen in oligodendrocytes, freshly isolated or in situ. Gap junctions were small and rare. Apposed oligodendrocyte plasma membrane formed linear tight junctions which became more numerous with time in culture. Thus, cultured oligodendrocytes isolated from ovine brains develop and maintain features characteristic of mature oligodendrocytes in situ and can be used to explore formation and maintenance of tight junctions and possibly other classes of cell-cell interactions important in the process of myelination.

Process defects and in situ monitoring methods in metal powder bed fusion: a review

International Nuclear Information System (INIS)

Grasso, Marco; Colosimo, Bianca Maria

2017-01-01

Despite continuous technological enhancements of metal Additive Manufacturing (AM) systems, the lack of process repeatability and stability still represents a barrier for the industrial breakthrough. The most relevant metal AM applications currently involve industrial sectors (e.g. aerospace and bio-medical) where defects avoidance is fundamental. Because of this, there is the need to develop novel in situ monitoring tools able to keep under control the stability of the process on a layer-by-layer basis, and to detect the onset of defects as soon as possible. On the one hand, AM systems must be equipped with in situ sensing devices able to measure relevant quantities during the process, a.k.a. process signatures. On the other hand, in-process data analytics and statistical monitoring techniques are required to detect and localize the defects in an automated way. This paper reviews the literature and the commercial tools for in situ monitoring of powder bed fusion (PBF) processes. It explores the different categories of defects and their main causes, the most relevant process signatures and the in situ sensing approaches proposed so far. Particular attention is devoted to the development of automated defect detection rules and the study of process control strategies, which represent two critical fields for the development of future smart PBF systems. (paper)

Process defects and in situ monitoring methods in metal powder bed fusion: a review

Science.gov (United States)

Grasso, Marco; Colosimo, Bianca Maria

2017-04-01

Despite continuous technological enhancements of metal Additive Manufacturing (AM) systems, the lack of process repeatability and stability still represents a barrier for the industrial breakthrough. The most relevant metal AM applications currently involve industrial sectors (e.g. aerospace and bio-medical) where defects avoidance is fundamental. Because of this, there is the need to develop novel in situ monitoring tools able to keep under control the stability of the process on a layer-by-layer basis, and to detect the onset of defects as soon as possible. On the one hand, AM systems must be equipped with in situ sensing devices able to measure relevant quantities during the process, a.k.a. process signatures. On the other hand, in-process data analytics and statistical monitoring techniques are required to detect and localize the defects in an automated way. This paper reviews the literature and the commercial tools for in situ monitoring of powder bed fusion (PBF) processes. It explores the different categories of defects and their main causes, the most relevant process signatures and the in situ sensing approaches proposed so far. Particular attention is devoted to the development of automated defect detection rules and the study of process control strategies, which represent two critical fields for the development of future smart PBF systems.

In situ stress measurement with the new LVDT - Cell - method description and verification

International Nuclear Information System (INIS)

Hakala, M.; Christiansson, R.; Martin, D.; Siren, T.; Kemppainen, K.

2013-11-01

Posiva Oy and SKB (Svensk Kaernbraenslehantering AB) tested the suitability a new LVDT-cell (Linear Variable Differential Transducer cell) to measure the induced stresses in the vicinity of an excavated surface and further to use these results to interpret the in situ state of stress. It utilises the overcoring methodology, measuring the radial convergence of four diameters using eight LVDTs, and is similar in concept to the USBM-gauge. A 127 mm diameter pilot-hole is required and the overcore diameter is 200 mm. The minimum overcoring length is 350 mm, and hence a compact drill can be utilised. Extensive testing of the LVDT-cell shows it to be robust and suitable for use in an underground environment. Sensitivity tests also show that the cell can withstand a range of operating conditions and still provide acceptable results. The in situ stress at the measurement location can be solved by numerical inversion using the results of at least three overcoring measurements around the three-dimensional tunnel section. The large dimensions of the measurement tool and the ability to utilise multiple measurements at various locations in a tunnel section, provides flexibility in selecting an appropriate rock mass volume. Because the inversion technique relies on knowing the exact location of the measurements and the geometry profile of the tunnel, modern survey techniques such as Lidar or photogrammetric technology should be used. Checks using traditional surveying techniques should also be used to ensure adequate survey resolution, specially in case of sidecoring measurements. To evaluate the suitability of the LVDT-cell to provide the in situ state of stress, tests were carried out in the drill-and-blast TASS tunnel and TBM tunnel at the Aespoe Hard Rock Laboratory in Sweden. The state of stress established using the LVDT-cell was in agreement with the state of stress established previously using traditional overcoring and hydraulic fracturing methods. In this study, the

In situ stress measurement with the new LVDT - Cell - method description and verification

Energy Technology Data Exchange (ETDEWEB)

Hakala, M. [KMS Hakala Oy, Nokia (Finland); Christiansson, R. [Svensk Kaernbraenslehantering AB, Stockholm (Sweden); Martin, D. [Univ. of Alberta, Edmonton (Canada); Siren, T.; Kemppainen, K.

2013-11-15

Posiva Oy and SKB (Svensk Kaernbraenslehantering AB) tested the suitability a new LVDT-cell (Linear Variable Differential Transducer cell) to measure the induced stresses in the vicinity of an excavated surface and further to use these results to interpret the in situ state of stress. It utilises the overcoring methodology, measuring the radial convergence of four diameters using eight LVDTs, and is similar in concept to the USBM-gauge. A 127 mm diameter pilot-hole is required and the overcore diameter is 200 mm. The minimum overcoring length is 350 mm, and hence a compact drill can be utilised. Extensive testing of the LVDT-cell shows it to be robust and suitable for use in an underground environment. Sensitivity tests also show that the cell can withstand a range of operating conditions and still provide acceptable results. The in situ stress at the measurement location can be solved by numerical inversion using the results of at least three overcoring measurements around the three-dimensional tunnel section. The large dimensions of the measurement tool and the ability to utilise multiple measurements at various locations in a tunnel section, provides flexibility in selecting an appropriate rock mass volume. Because the inversion technique relies on knowing the exact location of the measurements and the geometry profile of the tunnel, modern survey techniques such as Lidar or photogrammetric technology should be used. Checks using traditional surveying techniques should also be used to ensure adequate survey resolution, specially in case of sidecoring measurements. To evaluate the suitability of the LVDT-cell to provide the in situ state of stress, tests were carried out in the drill-and-blast TASS tunnel and TBM tunnel at the Aespoe Hard Rock Laboratory in Sweden. The state of stress established using the LVDT-cell was in agreement with the state of stress established previously using traditional overcoring and hydraulic fracturing methods. In this study, the

Electrochemical studies, in-situ and ex-situ characterizations of different manganese compounds electrodeposited in aerated solutions; Etudes electrochimiques, suivis in-situ et caracterisations ex-situ de divers composes de manganese electrodeposes dans des solutions aerees

Energy Technology Data Exchange (ETDEWEB)

Peulon, S.; Lacroix, A.; Chausse, A. [Univ. d' Evry-val-d' Essonne, Laboratoire Analyse et Modelisation pour la Biologie et l' Environnement (LAMBE CNRS UMR 8587), 91 - Evry (France); Larabi-Gruet, N. [CEA Saclay, Dept. de Physico-Chimie (DEN/DPC/SECR/L3MR), 91 - Gif sur Yvette (France)

2007-07-01

This work deals with the electrodeposition of manganese compounds. A systematic study of the synthesis experimental conditions has been carried out, and the obtained depositions have been characterized by different ex-situ analyses methods (XRD, FTIR, SEM). The in-situ measurements of mass increase with a quartz microbalance during the syntheses have allowed to estimate the growth mechanisms which are in agreement with the ex-situ characterizations. The cation has an important role in the nature of the electrodeposited compounds. In presence of sodium, a mixed lamellar compound Mn(III)/Mn(IV), the birnessite, is deposited, whereas in presence of potassium, bixbyite is formed (Mn{sub 2}O{sub 3}), these two compounds having a main role in the environment. The substrate can also influence the nature of the formed intermediary compounds. Little studied compounds such as feitkneichtite ({beta}-MnOOH) and groutite ({alpha}-MnOOH) have been revealed. (O.M.)

Identification of Mucorales isolates from soil using morphological and molecular methods

Directory of Open Access Journals (Sweden)

Ardeshir Ziaee

2016-03-01

Full Text Available Background and Purpose: Soil is the main habitat of saprophytic and pathogenic fungi. Mucormycetes are one of the most parts of soil fungi and certain members are among opportunistic fungi and can cause systemic fungal infections in immunocompromised patients. The majority of human and animal infections are caused by members of the genera Rhizopus, Mucor, Rhizomucor, Lichtheimia (Absidia, Cunninghamella and Mortierella. The objective of this research was to isolate and identify the main genera of Zygomycetes, using molecular assay and morphological features. Materials and Methods: A total of 340 soil samples were collected from different sites of seven public parks and 14 municipality districts in Isfahan. All samples were cultured on appropriate media and incubated at 27° C for 2 to 4 days, and then examined daily for visible fungal growth. PCR-RFLP method and macroscopic, microscopic and physiological characteristics were applied to identify fungal colonies. Results: Four hundred pure colonies belonging to six genera of Zygomycetes including Lichtheimia, Rhizopus, Rhizomucor, Mucor, Cunninghamella and Mortierella were identified. The genus Rhizopus (35.5% was the most frequent isolate, followed by Mucor (32.25% and Rhizomucor (27.5%. Conclusion: These finding may help us to understand about the importance of opportunistic fungi in public areas and the risk of exposure with immunocompromised persons.

Comparison of Total RNA Isolation Methods for Analysis of Immune-Related microRNAs in Market Milks.

Science.gov (United States)

Oh, Sangnam; Park, Mi Ri; Son, Seok Jun; Kim, Younghoon

2015-01-01

Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63℃ for 30 min)-, high temperature for short time (HTST, 75℃ for 15 s)-, and ultra heat treatment (UHT, 120-130℃ for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.

Contribution of the JRC Ispra to the intercomparison of analysis methods for seismically isolated nuclear structures

International Nuclear Information System (INIS)

Magonette, G.; Renda, V.

2002-01-01

Aim of the work done at JRC has been essentially to investigate the potentiality of the Pseudo-Dynamic (PsD) method to test structures incorporating anti-seismic protection devices based on materials with a strain-rate dependent behaviour. This is of relevant importance due to the interest to perform tests on large-scale mock-ups to assess the behaviour of realistic structure of civil engineering interest. Two specific typologies of protection have been analysed and tested at the European Laboratory for Structural Assessment (ELSA) of JRC Ispra. The first dealing with base isolation and the second with energy dissipation devices. In both cases the protection devices were based on high damping rubber material which is characterised by a moderate dependence from the strain rate of the application of the displacements. To validate a standard procedure to test base isolated structures by the PsD method, a collaboration was set up with the Italian Working Group on Seismic Isolation which includes the national research centre ENEA, the national electricity board ENEL, the industrial research centre ISMES and a manufacturer of isolators ALGA. In the framework of this collaboration it was decided to test at the ELSA laboratory a scaled 5-storey frame structure (provided by ENEL), isolated by means of high damping rubber bearings (HDRBs), which had been tested on the shaking table of ISMES. This experimental activity aimed to compare the results which can be obtained by means of the PsD testing technique with those which can be obtained by means of a truly-dynamic test on a shaking table. To validate a standard procedure to test structures incorporating energy dissipation devices, an international collaboration has been set up with Industries, Research Centres and Universities in the framework of a project partially funded by the European Commission through the General Directorate for Science and Technology. The obtained results show once more that the PsD method, when

[Comparison between conventional methods, ChromAgar Candida® and PCR method for the identification of Candida species in clinical isolates].

Science.gov (United States)

Estrada-Barraza, Deyanira; Dávalos Martínez, Arturo; Flores-Padilla, Luis; Mendoza-De Elias, Roberto; Sánchez-Vargas, Luis Octavio

2011-01-01

The increase in the incidence of yeast species causing fungemia in susceptible immunocompromised patients in the last two decades and the low sensitivity of conventional blood culture has led to the need to develop alternative approaches for the early detection and identification of causative species. The aim of this study was to compare the usefulness of molecular testing by the polymerase chain reaction (PCR) and conventional methods to identify clinical isolates of different species, using the ID32C ATB system (bioMérieux, France), chromogenic culture Chromagar Candida® (CHROMagar, France) and morphogenesis in corn meal agar. We studied 79 isolates, in which the most prevalent species using the system ID32C and PCR was C. albicans, followed by C. tropicalis, C. glabrata and C .krusei. PCR patterns obtained for the identification of clinical isolates were stable and consistent in the various independent studies and showed good reproducibility, concluding that PCR with species-specific primers that amplify genes ITS1 and ITS2 for rRNA or topoisomerase II primers is a very specific and sensitive method for the identification of C. glabrata, C. krusei, C. albicans, and with less specificity for C. tropicalis. Copyright © 2010 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

Estimation of the in situ degradation of the washout fraction of starch by using a modified in situ protocol and in vitro measurements

NARCIS (Netherlands)

Jonge, de L.H.; Laar, van H.; Dijkstra, J.

2015-01-01

The in situ degradation of the washout fraction of starch in six feed ingredients (i.e. barley, faba beans, maize, oats, peas and wheat) was studied by using a modified in situ protocol and in vitro measurements. In comparison with the washing machine method, the modified protocol comprises a milder

In situ high-pressure measurement of crystal solubility by using neutron diffraction

Science.gov (United States)

Chen, Ji; Hu, Qiwei; Fang, Leiming; He, Duanwei; Chen, Xiping; Xie, Lei; Chen, Bo; Li, Xin; Ni, Xiaolin; Fan, Cong; Liang, Akun

2018-05-01

Crystal solubility is one of the most important thermo-physical properties and plays a key role in industrial applications, fundamental science, and geoscientific research. However, high-pressure in situ measurements of crystal solubility remain very challenging. Here, we present a method involving high-pressure neutron diffraction for making high-precision in situ measurements of crystal solubility as a function of pressure over a wide range of pressures. For these experiments, we designed a piston-cylinder cell with a large chamber volume for high-pressure neutron diffraction. The solution pressures are continuously monitored in situ based on the equation of state of the sample crystal. The solubility at a high pressure can be obtained by applying a Rietveld quantitative multiphase analysis. To evaluate the proposed method, we measured the high-pressure solubility of NaCl in water up to 610 MPa. At a low pressure, the results are consistent with the previous results measured ex situ. At a higher pressure, more reliable data could be provided by using an in situ high-pressure neutron diffraction method.

Introducing a New Experimental Islet Transplantation Model using Biomimetic Hydrogel and a Simple High Yield Islet Isolation Technique.

Science.gov (United States)

Mohammadi Ayenehdeh, Jamal; Niknam, Bahareh; Hashemi, Seyed Mahmoud; Rahavi, Hossein; Rezaei, Nima; Soleimani, Masoud; Tajik, Nader

2017-07-01

Islet transplantation could be an ideal alternative treatment to insulin therapy for type 1 diabetes Mellitus (T1DM). This clinical and experimental field requires a model that covers problems such as requiring a large number of functional and viable islets, the optimal transplantation site, and the prevention of islet dispersion. Hence, the methods of choice for isolation of functional islets and transplantation are crucial. The present study has introduced an experimental model that overcomes some critical issues in islet transplantation, including in situ pancreas perfusion by digestive enzymes through common bile duct. In comparison with conventional methods, we inflated the pancreas in Petri dishes with only 1 ml collagenase type XI solution, which was followed by hand-picking isolation or Ficoll gradient separation to purify the islets. Then we used a hydrogel composite in which the islets were embedded and transplanted into the peritoneal cavity of the streptozotocin-induced diabetic C57BL/6 mice. As compared to the yield of the classical methods, in our modified technique, the mean yield of isolation was about 130-200 viable islets/mouse pancreas. In vitro glucose-mediated insulin secretion assay indicated an appropriate response in isolated islets. In addition, data from in vivo experiments revealed that the allograft remarkably maintained blood glucose levels under 400 mg/dl and hydrogel composite prevents the passage of immune cells. In the model presented here, the rapid islet isolation technique and the application of biomimetic hydrogel wrapping of islets could facilitate islet transplantation procedures.

paediatric ureteric calculi: in-situ extracorporeal shock wave lithotripsy

African Journals Online (AJOL)

Objective To evaluate prospectively the efficacy of in-situ extracorporeal shock wave lithotripsy (ESWL) in the treatment of ureteric calculi in the paediatric age group. Patients and Methods Twenty children (aged 2.2 16 years) with 22 ureteric stones were evaluated and treated with in-situ ESWL using the Dornier S lithotripter ...

Field O stars: formed in situ or as runaways?

Science.gov (United States)

Gvaramadze, V. V.; Weidner, C.; Kroupa, P.; Pflamm-Altenburg, J.

2012-08-01

A significant fraction of massive stars in the Milky Way and other galaxies are located far from star clusters and star-forming regions. It is known that some of these stars are runaways, i.e. possess high space velocities (determined through the proper motion and/or radial velocity measurements), and therefore most likely were formed in embedded clusters and then ejected into the field because of dynamical few-body interactions or binary-supernova explosions. However, there exists a group of field O stars whose runaway status is difficult to prove via direct proper motion measurements (e.g. in the Magellanic Clouds) or whose (measured) low space velocities and/or young ages appear to be incompatible with their large separation from known star clusters. The existence of this group led some authors to believe that field O stars can form in situ. Since the question of whether or not O stars can form in isolation is of crucial importance for star formation theory, it is important to thoroughly test candidates of such stars in order to improve the theory. In this paper, we examine the runaway status of the best candidates for isolated formation of massive stars in the Milky Way and the Magellanic Clouds by searching for bow shocks around them, by using the new reduction of the Hipparcos data, and by searching for stellar systems from which they could originate within their lifetimes. We show that most of the known O stars thought to have formed in isolation are instead very likely runaways. We show also that the field must contain a population of O stars whose low space velocities and/or young ages are in apparent contradiction to the large separation of these stars from their parent clusters and/or the ages of these clusters. These stars (the descendants of runaway massive binaries) cannot be traced back to their parent clusters and therefore can be mistakenly considered as having formed in situ. We argue also that some field O stars could be detected in optical

An improved method for isolation of RNA from bone

Directory of Open Access Journals (Sweden)

Carter Lauren E

2012-01-01

Full Text Available Abstract Background Bone physiology is increasingly appreciated as an important contributor to metabolic disorders such as type 2 diabetes. However, progress in understanding the role of bone in determining metabolic health is hampered by the well-described difficulty of obtaining high quality RNA from bone for gene expression analysis using the currently available approaches. Results We developed a simple approach to isolate bone RNA that combines pulverizing the bone and the phenol-guanidinium based RNA extraction in a single step while maintaining near-freezing temperatures. This single step method increases the yield of high quality RNA by eight-fold, with RNA integrity numbers ranging from 6.7 to 9.2. Conclusions Our streamlined approach substantially increases the yield of high-quality RNA from bone tissue while facilitating safe and efficient processing of multiple samples using readily available platforms. The RNA obtained from this method is suitable for use in gene expression analysis in real-time quantitative PCR, microarray, and next generation sequencing applications.

Atomic scissors: a new method of tracking the 5-bromo-2'-deoxyuridine-labeled DNA in situ.

Directory of Open Access Journals (Sweden)

Anna Ligasová

Full Text Available A new method of the light microscopy detection of BrdU-labeled DNA in situ is described. It is based on the oxidative attack at the deoxyribose moiety by copper(I in the presence of oxygen, which leads to the abstraction of hydrogen atom from deoxyribose culminating in the elimination of the nucleobase, scission of the nucleic-acid strand and formation of frequent gaps. The gaps allow the reaction of the antibodies with the commonly used markers of replication (e.g. 5-bromo-2'-deoxyuridine, which are otherwise masked. The method developed makes it possible to detect nuclear and mitochondrial DNA replication efficiently. In most cases, it does not inhibit effective protein detections and in addition enables simultaneous localization of newly-synthesized RNA. The alternative presently-used methods result in protein denaturation and/or extensive DNA cleavage followed by the DNA-bound proteins peeling off.

Actinobacillus pleuropneumoniae osteomyelitis in pigs demonstrated by fluorescent in situ hybridization

DEFF Research Database (Denmark)

Jensen, Tim Kåre; Boye, Mette; Hagedorn-Olsen, T.

1999-01-01

Necrotizing osteomyelitis and fibrinopurulent arthritis with isolation of Actinobacillus pleuropneumoniae serotype 2 is reported in two pigs from a herd with lameness and mild coughing problems among 8 to 12-week-old pigs. Application of fluorescent in situ hybridization targeting 16S ribosomal R......, in joints with arthritis, and in bone necroses including lysis of growth plate and suppurative inflammation in the adjacent trabecular metaphysis, thus demonstrating that well-known infections manifest new, unusual lesions....

Evaluation of in situ sulfate reduction as redox buffer capacity in groundwater flow path

International Nuclear Information System (INIS)

Ioka, Seiichiro; Iwatsuki, Teruki; Amano, Yuki; Furue, Ryoji

2007-01-01

For safety assessment of geological isolation, it is important to evaluate in situ redox buffer capacity in high-permeability zone as groundwater flow path. The study evaluated in situ sulfate reduction as redox buffer capacity in the conglomerate bedding in Toki Lignite-bearing Formation, which occurs at the lowest part of sedimentary rocks overlying basement granite. The bedding plays an important role as the main groundwater flow path. The result showed that in situ redox buffer capacity in the conglomerate bedding has been identified on first nine months, whereas in the following period the redox buffer capacity has not been identified for about fifteen months. This will be caused by the bedding became inappropriate for microbial survival as the organic matter which is needfuel for microbial activity was consumed. Thus, there will be limited redox buffer capacity in groundwater flow path even in formation including organic matter-bearing layer. (author)

Isolation of BAC Clones Containing Conserved Genes from Libraries of Three Distantly Related Moths: A Useful Resource for Comparative Genomics of Lepidoptera

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Yuji Yasukochi

2011-01-01

Full Text Available Lepidoptera, butterflies and moths, is the second largest animal order and includes numerous agricultural pests. To facilitate comparative genomics in Lepidoptera, we isolated BAC clones containing conserved and putative single-copy genes from libraries of three pests, Heliothis virescens, Ostrinia nubilalis, and Plutella xylostella, harboring the haploid chromosome number, =31, which are not closely related with each other or with the silkworm, Bombyx mori, (=28, the sequenced model lepidopteran. A total of 108–184 clones representing 101–182 conserved genes were isolated for each species. For 79 genes, clones were isolated from more than two species, which will be useful as common markers for analysis using fluorescence in situ hybridization (FISH, as well as for comparison of genome sequence among multiple species. The PCR-based clone isolation method presented here is applicable to species which lack a sequenced genome but have a significant collection of cDNA or EST sequences.

Electrochemical studies, in-situ and ex-situ characterizations of different manganese compounds electrodeposited in aerated solutions

International Nuclear Information System (INIS)

Peulon, S.; Lacroix, A.; Chausse, A.; Larabi-Gruet, N.

2007-01-01

This work deals with the electrodeposition of manganese compounds. A systematic study of the synthesis experimental conditions has been carried out, and the obtained depositions have been characterized by different ex-situ analyses methods (XRD, FTIR, SEM). The in-situ measurements of mass increase with a quartz microbalance during the syntheses have allowed to estimate the growth mechanisms which are in agreement with the ex-situ characterizations. The cation has an important role in the nature of the electrodeposited compounds. In presence of sodium, a mixed lamellar compound Mn(III)/Mn(IV), the birnessite, is deposited, whereas in presence of potassium, bixbyite is formed (Mn 2 O 3 ), these two compounds having a main role in the environment. The substrate can also influence the nature of the formed intermediary compounds. Little studied compounds such as feitkneichtite (β-MnOOH) and groutite (α-MnOOH) have been revealed. (O.M.)

Basalt waste isolation project. Quarterly report, April 1, 1981-June 30, 1981

Energy Technology Data Exchange (ETDEWEB)

Deju, R.A.

1981-08-01

This document reports progress made in the Basalt Waste Isolation Project during the third quarter of fiscal year 1981. Efforts are described for the following programs of the project work breakdown structure: systems; waste package; site; repository; regulatory and institutional; test facilities; in situ test facilities.

Comparative studies on the interactions of the natural isolate KS5 and one reference strain (DSM 10134) with uranium(VI)

Energy Technology Data Exchange (ETDEWEB)

Gerber, Ulrike; Krawczyk-Baersch, Evelyn [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Div. Biogeochemistry; Arnold, Thuro [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Inst. of Resource Ecology

2016-07-01

The yeast KS5 (Rhodosporidium toruloides) was isolated by culture dependent method directly from the flooding water in Koenigstein (Germany). To compare the U tolerance and immobilization ability of the isolate a reference culture DSM 10134 (R. toruloides) was applied. Both cultures displayed the ability to tolerate high amounts of U, in contrast the reference KS5 showed a six-fold higher U tolerance in comparison to the reference strain. U immobilization studies displayed that both organisms are able to remove high amounts. The flooding water in Koenigstein has to be cleaned up for many years by an intensive waste water treatment plant. Possibly, with the help of natural occuring microorganisms the flooding water could be cleaned up using in situ bioremediation.

In situ permeability testing of rock salt

International Nuclear Information System (INIS)

Peterson, E.W.; Lagus, P.L.; Broce, R.D.; Lie, K.

1981-04-01

Storage of transuranic (TRU) wastes in bedded salt formations requires a knowledge of the in situ permeability of SENM rock salt. Since assumptions for safety assessments have been made in which these wastes could generate gas pressures on the order of the lithostatic pressure over geologic time scales, the permeability of the surrounding formation becomes an important parameter for determining the manner in which the gases will be contained or dispersed. This report describes the series of tests conducted in the AEC-7 borehole, located near the WIPP site, to determine the in situ gas flow characteristics of the bedded salt. In these tests, compressed air was injected into the borehole and flow into the surrounding formation measured. These measured flow rates were interpreted in terms of formation permeabilities and porosities which were, in turn, used as modeling parameters for the repository response analysis. Two series of field tests were performed. The first series consisted of a number of whole-hole flow tests conducted to provide preliminary design information required for future operation of a guarded straddle packer system capable of measuring permeabilities > or = 0.1 μdarcy. The second series of tests were conducted using the Systems, Science and Software (S-Cubed) designed guarded straddle packer system. In these interval permeability tests, 100-foot lengths of borehole were isolated and the flow characteristics of the surrounding formation examined. In this report, a complete description of the test procedures, instrumentation, and measurement techniques is first given. The analytical/numerical methods used for data interpretation are then presented, followed by results of the interval and permeability tests. (The whole-hole tests are summarized in Appendix A.) Conclusions are presented in the final section

[Isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris in Yunnan Safari Park].

Science.gov (United States)

Cao, Yanru; Jiang, Yi; Li, Youlong; Chen, Xiu; Jin, Rongxian; He, Wenxiang

2012-07-04

We studied the isolation methods and diversity of culturable fecal actinobacteria associated with Panthera tigris tigris by using culture-dependent approaches. Fresh fecal samples of healthy Panthera tigris tigris were collected from Yunnan Safari Park. Pretreatment of the samples, isolation media and inhibitors were tested for actinobacteria isolation. 16S rRNA genes of actinobacteria were sequenced and subjected to phylogenetic analysis. The abundance of culturable actinobacteria was 1.10 x 10(8) cfu/g colony forming units (CFU) per gram of feces (wet weight). We obtained 110 purified cultural actinobacterium strains. The analysis based on 16S rRNA gene sequences showed that these strains were distributed in 10 different families and 12 genera of actinobacteria at least, and most of them were non-filamentous, such as Arthrobacter, Dietzia, Kocuria, Corynebacterium and Microbacterium. Streptomyces was the mainly classical filamentous actinobacteria, and up to 64% of total. Drying and heating up the fecal samples can greatly increase the rate of the actinobacteria. Many kinds of inhibitors and chemical defined media are suitable for isolation of fecal actinobacteria. The culturable actinobacteria are abundant in Panthera tigris tigris feces. Our study found an effective method to isolate animals' fecal actinobacteria and it's useful for studying and exploiting animals' fecal actinobacteria.

In Situ Dark Adaptation Enhances the Efficiency of DNA Extraction from Mature Pin Oak (Quercus palustris Leaves, Facilitating the Identification of Partial Sequences of the 18S rRNA and Isoprene Synthase (IspS Genes

Directory of Open Access Journals (Sweden)

Csengele E. Barta

2017-10-01

Full Text Available Mature oak (Quercus spp. leaves, although abundantly available during the plants’ developmental cycle, are rarely exploited as viable sources of genomic DNA. These leaves are rich in metabolites difficult to remove during standard DNA purification, interfering with downstream molecular genetics applications. The current work assessed whether in situ dark adaptation, to deplete sugar reserves and inhibit secondary metabolite synthesis could compensate for the difficulties encountered when isolating DNA from mature leaves rich in secondary metabolites. We optimized a rapid, commercial kit based method to extract genomic DNA from dark- and light-adapted leaves. We demonstrated that in situ dark adaptation increases the yield and quality of genomic DNA obtained from mature oak leaves, yielding templates of sufficiently high quality for direct downstream applications, such as PCR amplification and gene identification. The quality of templates isolated from dark-adapted pin oak leaves particularly improved the amplification of larger fragments in our experiments. From DNA extracts prepared with our optimized method, we identified for the first time partial segments of the genes encoding 18S rRNA and isoprene synthase (IspS from pin oak (Quercus palustris, whose full genome has not yet been sequenced.

Isolation and Identification of Volatile Components in Tempe by Simultaneous Distillation-Extraction Method by Modified Extraction Method

Directory of Open Access Journals (Sweden)

Syahrial Syahrial

2010-06-01

Full Text Available An isolation and identification of volatile components in temps for 2, 5 and 8 days fermentation by simultaneous distillation-extraction method was carried out. Simultaneous distillation-extraction apparatus was modified by Muchalal from the basic Likens-Nickerson's design. Steam distillation and benzena as an extraction solvent was used in this system. The isolation was continuously carried out for 3 hours which maximum water temperature In the Liebig condenser was 8 °C. The extract was concentrated by freeze concentration method, and the volatile components were analyzed and identified by combined gas chromatography-mass spectrophotometry (GC-MS. The Muchalal's simultaneous distillation extraction apparatus have some disadvantage in cold finger condenser, and it's extractor did not have condenser. At least 47, 13 and 5 volatile components were found in 2, 5 and 8 days fermentation, respectively. The volatile components in the 2 days fermentation were nonalal, ɑ-pinene, 2,4-decadienal, 5-phenyldecane, 5-phenylundecane, 4-phenylundecane, 5-phenyldodecane, 4-phenyldodecane, 3-phenyldodecane, 2-phenyldodecane, 5-phenyltridecane, and caryophyllene; in the 5 days fermentation were nonalal, caryophyllene, 4-phenylundecane, 5-phenyldodecane, 4-phenyldodecane, 3-phenyldodecane, 2-phenyldodecane; and in the 8 days fermentation were ethenyl butanoic, 2-methy1-3-(methylethenylciclohexyl etanoic and 3,7-dimethyl-5-octenyl etanoic.

Biodegradation Capability of Some Bacteria Isolates to Use Lubricant Oil in Vitro

Science.gov (United States)

Ahda, Y.; Azhar, M.; Fitri, L.; Afnida, A.; Adha, G. S.; Alifa, W. N.; Handayani, D.; Putri, D. H.; Irdawati, I.; Chatri, M.

2018-04-01

Our previous study identified three species of bacteria, i.e. Alcaligenes sp., Bacillus spl, and Bacillus sp2 isolated from using lubricant oil-contaminated soil in a Padang’s workshop. However, its ability to degrade hydrocarbon were not known yet. In this extension study, we explore a wider area to find more hydrocarbonoclastic bacteria and examined its capability to degrade hydrocarbon in vitro. Seventeen isolates were characterized its capability using NA + used lubricant oil + tween + neutral red medium. Isolates A1, B2, D1 and D4 shows the high degradation index, whereas isolates A2, A3, A5, D2, B1, B3 and isolates A4, B4, D3 have medium and low degradation index, respectively. These potential hydrocarbonoclastic bacteria need in situ characterization to know their actual activities for bioremediation.

Temperature-dependency analysis and correction methods of in-situ power-loss estimation for crystalline silicon modules undergoing potential-induced degradation stress testing

DEFF Research Database (Denmark)

Spataru, Sergiu; Hacke, Peter; Sera, Dezso

2015-01-01

We propose a method of in-situ characterization of the photovoltaic module power at standard test conditions using superposition of the dark current-voltage (I-V) curve measured at elevated stress temperature during potential-induced degradation (PID) testing. PID chamber studies were performed o...

Characterization of RNA from Exosomes and Other Extracellular Vesicles Isolated by a Novel Spin Column-Based Method

Science.gov (United States)

Enderle, Daniel; Spiel, Alexandra; Coticchia, Christine M.; Berghoff, Emily; Mueller, Romy; Schlumpberger, Martin; Sprenger-Haussels, Markus; Shaffer, Jonathan M.; Lader, Eric; Skog, Johan; Noerholm, Mikkel

2015-01-01

Exosomes and other extracellular vesicles (commonly referred to as EVs) have generated a lot of attention for their potential applications in both diagnostics and therapeutics. The contents of these vesicles are the subject of intense research, and the relatively recent discovery of RNA inside EVs has raised interest in the biological function of these RNAs as well as their potential as biomarkers for cancer and other diseases. Traditional ultracentrifugation-based protocols to isolate EVs are labor-intensive and subject to significant variability. Various attempts to develop methods with robust, reproducible performance have not yet been completely successful. Here, we report the development and characterization of a spin column-based method for the isolation of total RNA from EVs in serum and plasma. This method isolates highly pure RNA of equal or higher quantity compared to ultracentrifugation, with high specificity for vesicular over non-vesicular RNA. The spin columns have a capacity to handle up to 4 mL sample volume, enabling detection of low-abundance transcripts in serum and plasma. We conclude that the method is an improvement over traditional methods in providing a faster, more standardized way to achieve reliable high quality RNA preparations from EVs in biofluids such as serum and plasma. The first kit utilizing this new method has recently been made available by Qiagen as “exoRNeasy Serum/Plasma Maxi Kit”. PMID:26317354

In-Situ Leak Testing And Replacement Of Glovebox Isolator, Or Containment Unit Gloves

Science.gov (United States)

Castro, Julio M.; Macdonald, John M.; Steckle, Jr., Warren P.

2004-11-02

A test plug for in-situ testing a glove installed in a glovebox is provided that uses a top plate and a base plate, and a diametrically expandable sealing mechanism fitting between the two plates. The sealing mechanism engages the base plate to diametrically expand when the variable distance between the top plate and the bottom plate is reduced. An inlet valve included on the top plate is used to introducing a pressurized gas to the interior of the glove, and a pressure gauge located on the top plate is used to monitor the interior glove pressure.

An adhesion-based method for plasma membrane isolation: evaluating cholesterol extraction from cells and their membranes.

Science.gov (United States)

Bezrukov, Ludmila; Blank, Paul S; Polozov, Ivan V; Zimmerberg, Joshua

2009-11-15

A method to isolate large quantities of directly accessible plasma membrane from attached cells is presented. The method is based on the adhesion of cells to an adsorbed layer of polylysine on glass plates, followed by hypotonic lysis with ice-cold distilled water and subsequent washing steps. Optimal conditions for coating glass plates and time for cell attachment were established. No additional chemical or mechanical treatments were used. Contamination of the isolated plasma membrane by cell organelles was less than 5%. The method uses inexpensive, commercially available polylysine and reusable glass plates. Plasma membrane preparations can be made in 15 min. Using this method, we determined that methyl-beta-cyclodextrin differentially extracts cholesterol from fibroblast cells and their plasma membranes and that these differences are temperature dependent. Determination of the cholesterol/phospholipid ratio from intact cells does not reflect methyl-beta-cyclodextrin plasma membrane extraction properties.

In situ solution mining technique

International Nuclear Information System (INIS)

Learmont, R.P.

1978-01-01

A method of in situ solution mining is disclosed in which a primary leaching process employing an array of 5-spot leaching patterns of production and injection wells is converted to a different pattern by converting to injection wells all the production wells in alternate rows

Comparative study of in situ methods for potential and actual evapotranspiration determination and their calculation by simulation model

International Nuclear Information System (INIS)

Kolev, B.

2006-01-01

Four in situ methods for potential and actual evapotranspiration determining were compared: neutron gauge, tensiometers, gypsum blocks and lysimeters. The actual and potential evapotranspiration were calculated by water balance equation and by using a simulation model for their determination. The aim of this study was mainly pointed on calculations of water use efficiency and transpiration coefficient in potential production situation. This makes possible to choose the best way for water consumption optimization for a given crop. The final results find with the best of the methods could be used for applying the principles of sustainable agricultural production in random object of Bulgarian agricultural area

A highly efficient nonchemical method for isolating live nematodes (Caenorhabditis elegans) from soil during toxicity assays.

Science.gov (United States)

Kim, Shin Woong; Moon, Jongmin; An, Youn-Joo

2015-01-01

The success of soil toxicity tests using Caenorhabditis elegans may depend in large part on recovering the organisms from the soil. However, it can be difficult to learn the International Organization for Standardization/ASTM International recovery process that uses the colloidal silica flotation method. The present study determined that a soil-agar isolation method provides a highly efficient and less technically demanding alternative to the colloidal silica flotation method. Test soil containing C. elegans was arranged on an agar plate in a donut shape, a linear shape, or a C curve; and microbial food was placed outside the soil to encourage the nematodes to leave the soil. The effects of ventilation and the presence of food on nematode recovery were tested to determine the optimal conditions for recovery. A linear arrangement of soil on an agar plate that was sprinkled with microbial food produced nearly 83% and 90% recovery of live nematodes over a 3-h and a 24-h period, respectively, without subjecting the nematodes to chemical stress. The method was tested using copper (II) chloride dihydrate, and the resulting recovery rate was comparable to that obtained using colloidal silica flotation. The soil-agar isolation method portrayed in the present study enables live nematodes to be isolated with minimal additional physicochemical stress, making it a valuable option for use in subsequent sublethal tests where live nematodes are required. © 2014 SETAC.

Oral rinse as a potential method to culture Candida isolate from AIDS patients

Directory of Open Access Journals (Sweden)

Desiana Radithia

2011-12-01

Full Text Available Background: Candida isolate is easily sampled from oral cavity by swabbing directly on the candidiasis lesion, to be smeared onto slides for direct examination or cultured in a growth medium. This method is by far the gold standard for defining candidiasis diagnosis. However it is difficult to apply on sensitive patients and almost impossible on patients showing no clinical appearance of oral candidiasis. AIDS patients are very prone to candida infection and have a tendency of repetitive infection involving mixed species. As many candida species show different susceptibility to anti-fungal agents, it is necessary to identify the species causing the infection in the management of oral candidiasis. Oral rinse is a suggested method to obtain candida isolate to be cultured for further analysis such as species identification. This method is simple and less risky on infection transmission as less tools are required in the procedure. Purpose: This study aimed to assess the application of oral rinse as an alternative method to culture Candida isolate from AIDS patients. Methods: A cross-sectional observative study was conducted in HIV/AIDS in-Patient Facility of Intermediate Care Unit for Infection Disease, Dr. Soetomo Hospital Surabaya. Fourteen stadium 4 AIDS patients matching criteria were swabbed on 1/3-posterior of the tongue, and then given 10 ml phosphate buffer saline to rinse vigorously for 15 seconds. Both specimens were cultured on Sabouraud’s dextrose agar and colony growth was observed. Results: Candida colonies were able to grow from all 14 isolates (100% by both methods. Qualitatively, cultures from oral rinse specimens were more populated than cultures from swab specimens. Conclusion: Oral rinse is an applicable technique to obtain Candida species isolate. This technique is safe, easy, non-invasive, and needs less tools therefore less risky for HIV transmission.Latar belakang: Isolat Candida mudah diambil dengan cara mengusap lesi

A fast and reproducible method for albumin isolation and depletion from serum and cerebrospinal fluid.

Science.gov (United States)

Holewinski, Ronald J; Jin, Zhicheng; Powell, Matthew J; Maust, Matthew D; Van Eyk, Jennifer E

2013-03-01

Analysis of serum and plasma proteomes is a common approach for biomarker discovery, and the removal of high-abundant proteins, such as albumin and immunoglobins, is usually the first step in the analysis. However, albumin binds peptides and proteins, which raises concerns as to how the removal of albumin could impact the outcome of the biomarker study while ignoring the possibility that this could be a biomarker subproteome itself. The first goal of this study was to test a new commercially available affinity capture reagent from Protea Biosciences and to compare the efficiency and reproducibility to four other commercially available albumin depletion methods. The second goal of this study was to determine if there is a highly efficient albumin depletion/isolation system that minimizes sample handling and would be suitable for large numbers of samples. Two of the methods tested (Sigma and ProteaPrep) showed an albumin depletion efficiency of 97% or greater for both serum and cerebrospinal fluid (CSF). Isolated serum and CSF albuminomes from ProteaPrep spin columns were analyzed directly by LC-MS/MS, identifying 128 serum (45 not previously reported) and 94 CSF albuminome proteins (17 unique to the CSF albuminome). Serum albuminome was also isolated using Vivapure anti-HSA columns for comparison, identifying 105 proteins, 81 of which overlapped with the ProteaPrep method. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Determination of chlorophenols in honey samples using in-situ ionic liquid-dispersive liquid-liquid microextraction as a pretreatment method followed by high-performance liquid chromatography.

Science.gov (United States)

Fan, Chen; Li, Nai; Cao, Xueli

2015-05-01

In-situ ionic liquid-dispersive liquid-liquid microextraction (IL-DLLME) method was developed as a pretreatment method for the detection of six chlorophenols (CPs) in honey samples. The hydrophobic ionic liquid [C4MIM][NTf2], formed in-situ by the hydrophilic ionic liquid [C4MIM][BF4] and the ion exchange reagent LiNTf2 was used as the microextractant solvent of CPs from honey sample. Then the enriched analytes were back-extracted into 40 μL of 0.14 M NaOH solution and finally subjected to analysis by high-performance liquid chromatography. The method showed low limit of detection of CPs, 0.8-3.2 μg/L and high enrichment factor, 34-65 with the recoveries range from 91.60% to 114.33%. The method is simple, rapid, environmentally friendly and with high extraction efficiency. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Comparison of biochemical and molecular methods for the identification of bacterial isolates associated with failed loggerhead sea turtle eggs.

Science.gov (United States)

Awong-Taylor, J; Craven, K S; Griffiths, L; Bass, C; Muscarella, M

2008-05-01

Comparison of biochemical vs molecular methods for identification of microbial populations associated with failed loggerhead turtle eggs. Two biochemical (API and Microgen) and one molecular methods (16s rRNA analysis) were compared in the areas of cost, identification, corroboration of data with other methods, ease of use, resources and software. The molecular method was costly and identified only 66% of the isolates tested compared with 74% for API. A 74% discrepancy in identifications occurred between API and 16s rRNA analysis. The two biochemical methods were comparable in cost, but Microgen was easier to use and yielded the lowest discrepancy among identifications (29%) when compared with both API 20 enteric (API 20E) and API 20 nonenteric (API 20NE) combined. A comparison of API 20E and API 20NE indicated an 83% discrepancy between the two methods. The Microgen identification system appears to be better suited than API or 16s rRNA analysis for identification of environmental isolates associated with failed loggerhead eggs. Most identification methods are not intended for use with environmental isolates. A comparison of identification systems would provide better options for identifying environmental bacteria for ecological studies.

Gas injection to inhibit migration during an in situ heat treatment process

Science.gov (United States)

Kuhlman, Myron Ira; Vinegar; Harold J.; Baker, Ralph Sterman; Heron, Goren

2010-11-30

Methods of treating a subsurface formation are described herein. Methods for treating a subsurface treatment area in a formation may include introducing a fluid into the formation from a plurality of wells offset from a treatment area of an in situ heat treatment process to inhibit outward migration of formation fluid from the in situ heat treatment process.

Phene Plate (PhP) biochemical fingerprinting. A screening method for epidemiological typing of enterococcal isolates.

Science.gov (United States)

Saeedi, B; Tärnberg, M; Gill, H; Hällgren, A; Jonasson, J; Nilsson, L E; Isaksson, B; Kühn, I; Hanberger, H

2005-09-01

Pulsed-field gel electrophoresis (PFGE) is currently considered the gold standard for genotyping of enterococci. However, PFGE is both expensive and time-consuming. The purpose of this study was to investigate whether the PhP system can be used as a reliable clinical screening method for detection of genetically related isolates of enterococci. If so, it should be possible to minimize the number of isolates subjected to PFGE typing, which would save time and money. Ninety-nine clinical enterococcal isolates were analysed by PhP (similarity levels 0.90-0.975) and PFGE (similarity levels PhP also belong to the same cluster according to PFGE, i.e. p(A(PFGE)=B(PFGE) * A(PhP)=B(PhP)), and the probability that a pair of isolates of different types according to PhP also belong to different clusters according to PFGE, i.e. p(A(PFGE) not equalB(PFGE) * A(PhP) not equalB(PhP)), was relatively high for E. faecalis (0.86 and 0.96, respectively), but was lower for E. faecium (0.51 and 0.77, respectively). The concordance which shows the probability that PhP and PFGE agree on match or mismatch was 86%-93% for E. faecalis and 54%-66% for E. faecium, which indicates that the PhP method may be useful for epidemiological typing of E. faecalis in the current settings but not for E. faecium.

Comparison of Plasmagel with LeucoPREP-Macrodex methods for separation of leukocytes for virus isolation.

Science.gov (United States)

Woods, G L; Proffitt, M R

1987-10-01

Plasmagel (Cellular Products, Inc., Buffalo, NY), which can separate both polymorphonuclear leukocytes (PMN) and mononuclear cells from other blood components, and LeucoPREP (Becton Dickinson Immunocytometry Systems, Mountain View, CA), which can separate mononuclear cells from other blood components, were used to harvest leukocytes from whole blood for the purpose of virus isolation. Macrodex was combined with the later, in a second step, for recovery of PMN. Of 90 peripheral blood specimens examined, cytomegalovirus was recovered from 10: in six by both methods, in three from Plasmagel prepared cells only, and in one from cells from the LeucoPREP-Macrodex preparation only. Total leukocyte counts, differential counts, and leukocyte viability did not differ significantly for the two methods. Plasmagel provided an efficient, inexpensive means of harvesting leukocytes from whole blood for virus isolation.

The theory and method of two-well field test for in-situ leaching uranium

International Nuclear Information System (INIS)

Yao Yixuan; Huo Jiandang; Xiang Qiulin; Tang Baobin

2007-01-01

Because leaching area in field test for in-situ leaching uranium is not accounted exactly, the reliability of obtaining parameters by calculating can not be ensured, and the whole test needs a long time and great investment. In two-well field test, lixiviant is injected from one well, pregnant solution is pumped out from the other, flow rate of the production well is more than that of the injection well, and uranium is not recoveried. In the case of keeping invariable ratio of pumping capacity to injecting capacity during the testing process, leaching area is not variable, can be exactly calculated. The full field test needs six months to one year. Two-well test is a scientific, rapid, minimal spending field test method, and is widely used in Commonwealth of Independent States. (authors)

Analysis of in situ stress at Yucca Mountain

International Nuclear Information System (INIS)

Bauer, S.J.; Holland, J.F.

1987-01-01

A method has been developed to initialize far-field finite element models such that the measured in situ stress state appears to be reproduced well. The method includes use of the mechanical stratigraphy, mechanical effect of pore pressure, gravity loading, a horizontal ''tectonic'' component of stress, and use of a jointed rock model to calculate the mechanical response. Topographic effects and effects related to the vertical variation in mechanical properties are predicted for repository depths (∼ 300 m). Gravity loading with a small horizontal compression is used to calculate a minimum horizontal stress similar in magnitude to that measured in situ. 8 refs., 5 figs

Automated gas bubble imaging at sea floor – a new method of in situ gas flux quantification

Directory of Open Access Journals (Sweden)

G. Bohrmann

2010-06-01

Full Text Available Photo-optical systems are common in marine sciences and have been extensively used in coastal and deep-sea research. However, due to technical limitations in the past photo images had to be processed manually or semi-automatically. Recent advances in technology have rapidly improved image recording, storage and processing capabilities which are used in a new concept of automated in situ gas quantification by photo-optical detection. The design for an in situ high-speed image acquisition and automated data processing system is reported ("Bubblemeter". New strategies have been followed with regards to back-light illumination, bubble extraction, automated image processing and data management. This paper presents the design of the novel method, its validation procedures and calibration experiments. The system will be positioned and recovered from the sea floor using a remotely operated vehicle (ROV. It is able to measure bubble flux rates up to 10 L/min with a maximum error of 33% for worst case conditions. The Bubblemeter has been successfully deployed at a water depth of 1023 m at the Makran accretionary prism offshore Pakistan during a research expedition with R/V Meteor in November 2007.

Identification of Candida albicans and Candida dubliniensis Species Isolated from Bronchoalveolar Lavage Samples Using Genotypic and Phenotypic Methods

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Sahar Kianipour

2018-01-01

Full Text Available Background: Candida dubliniensis is a newly diagnosed species very similar to Candida albicans phenotypically and first discovered in the mouth of people with AIDS in 1995. Among the different phenotypic and genotypic methods, a cost-effective method should be selected which makes it possible to differentiate these similar species. Materials and Methods: Polymerase chain reaction (PCR-restriction fragment length polymorphism with MspI enzyme and the Duplex-PCR method were done by DNA extraction using boiling. The sequencing of the amplified ribosomal region was used to confirm the C. dubliniensis species. Direct examination and colony count of the yeasts were applied for bronchoalveolar lavage (BAL samples and the growth rate of the yeasts were studied at 45°C. To understand the ability formation of chlamydoconidia in yeast isolates, they were separately cultured on the sunflower seed agar, wheat flour agar, and corn meal agar media. Results: Fifty-nine (49.2% yeast colonies were identified from the total of 120 BAL specimens. Twenty-nine isolated yeasts; including 17 (58.6% of C. albicans/dubliniensis complex and 12 (41.4% of nonalbicans isolates produced pseudohypha or blastoconidia in direct smear with a mean colony count of 42000 CFU/mL. C. albicans with the frequency of 15 (42.9% were the most common isolated yeasts, whereas C. dubliniensis was identified in two nonHIV patients. Conclusion: Sequencing of the replicated gene fragment is the best method for identifying the yeasts, but the determination of the species by phenotypic methods such as the creation of chlamydoconidia in sunflower seeds agar and wheat flour agar media can be cost-effective, have sensitivity and acceptable quality.

An Overview of Neolignans of the Genus Piper L.: Isolation Methods and Biological Activities.

Science.gov (United States)

Macedo, Arthur Ladeira; Dos Santos, Thais Carvalho Costa; Valverde, Alessandra Leda; Moreira, Davyson de Lima; Vasconcelos, Thatyana Rocha Alves

2017-01-01

The genus Piper L. has the shikimic acid pathway predominantly expressed, biosynthesizing many cinnamic acid derivatives (CAD). Neolignans comprise an important class of CAD that exhibit a wide range of pharmacological properties such as antibacterial, antitumor, insecticidal, anti-inflammatory, antioxidant, smooth muscle relaxant, neuroprotective, antiprotozoal and against platelet aggregation factor. These substances have been extracted and isolated from Piper species using different technics. The present review aims to summarize extraction and isolation methods and biological activities of the different types of neolignans covering the period from 1968 to January 2016. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Recent developments for in situ treatment of metal contaminated soils

Energy Technology Data Exchange (ETDEWEB)

NONE

1997-03-01

Metals contamination is a common problem at hazardous waste sites. This report assists the remedy selection process by providing information on four in situ technologies for treating soil contaminated with metals. The four approaches are electrokinetic remediation, phytoremediation, soil flushing, and solidification/stabilization. Electrokinetic remediation separates contaminants from soil through selective migration upon application of an electric current. Phytoremediation is an emerging technology that uses plants to isolate or stabilize contaminants. Soil flushing techniques promote mobility and migration of metals by solubilizing contaminants so that they can be recovered. Two types of in situ solidification/stabilization (S/S) techniques are discussed, one based on addition of reagents and the other based on the use of energy. The report discusses different techniques currently in practice or under development, identifies vendors and summarizes performance data, and discusses technology attributes that should be considered during early screening of potential remedies. 8 refs., 9 figs., 9 tabs., 2 apps.

A method to determine insulin responsiveness in synaptosomes isolated from frozen brain tissue.

Science.gov (United States)

Franklin, Whitney; Taglialatela, Giulio

2016-03-01

Studying the insulin signaling response at the synapse is an important approach to understand molecular mechanisms involved in disease-related neurodegenerative processes. We developed a method for studying the insulin responsiveness at the synaptic level by isolating functional synaptosomes from fresh or frozen tissue and exposing them to insulin in the presence of ATP (a critical step) to detect insulin receptor (IR) activation. We performed an ATP dose-response curve, insulin dose-response curve, and insulin response time course to optimize this method. We also demonstrated that our protocol reflects the degree of insulin responsiveness in vivo by using an animal model of known insulin resistance, AtENPP1-Tg mice. This method is advantageous over other methods detecting IR in total brain homogenates due to the ability to detect IR response without confounding contributions from other cell areas and cell types also expressing IR. Furthermore, ex vivo insulin stimulation can be compared to baseline synaptosomes obtained from the same animal which improves reliability and statistical power while decreasing the number of animals required to perform individual experiments. We have developed a reliable, efficient method to measure insulin-driven ex vivo phosphorylation of the synaptosomal insulin receptor that can reliably reflect the pre-existing insulin responsiveness status in the CNS of the animal. To the best of our knowledge, this is the first evidence of stimulation of isolated synaptosomes with insulin and a promising new technique to study the synaptic CNS insulin responsiveness under physiological or disease conditions. Copyright © 2016 Elsevier B.V. All rights reserved.

A Modified Ficoll-Paque Gradient Method for Isolating Mononuclear Cells from the Peripheral and Umbilical Cord Blood of Humans for Biobanks and Clinical Laboratories.

Science.gov (United States)

Jia, Yanjuan; Xu, Hui; Li, Yonghong; Wei, Chaojun; Guo, Rui; Wang, Fang; Wu, Yu; Liu, Jing; Jia, Jing; Yan, Junwen; Qi, Xiaoming; Li, Yuanting; Gao, Xiaoling

2018-04-01

Although the Ficoll-Paque method is classically used to isolate peripheral blood mononuclear cells (PBMCs), modifications in this method are required for a more rapid and economic output for biobanks and clinical laboratories, particularly in developing countries. In this study, we addressed this issue by modifying the Ficoll-Paque method for the isolation of PBMCs or mononuclear cells from the peripheral and the umbilical cord blood of healthy and diseased (infected, anemic, and chronic obstructive pulmonary disease) adult individuals. In the modified method, we initiated the cell isolation process from the buffy coat layer, which appears in the interface between the plasma and sediments after centrifugation, instead of using the whole blood as described in the classic method. Although the PBMC yield by the modified method was about 12% less than in the classic method, the number of PBMCs isolated by the modified method was more than one million, which is enough for different research/diagnostic purposes, such as multi-omics detection. Assessment of cell viability and purity by hematology analyzer and trypan blue showed no significant difference between the viability and purity of the PBMCs isolated by these two methods in almost all groups, except samples from the infected and cord blood groups, where lower PBMC purity with higher granulocyte contamination were observed. In addition, at delayed processing time points, all parameters for the two methods were decreased in a time-dependent manner, especially at 8, 12, or 24 hours after the sample collection. In summary, the performance of PBMC isolation by the classic and modified methods mainly relies on the PBMC ratio in original samples. The modified method could be preferred for PBMC isolation because of its time and cost savings, especially for the biobanks and clinical laboratories in developing countries.

A comparison of three methods for the isolation of Arcobacter spp. from retail raw poultry in Northern Ireland

DEFF Research Database (Denmark)

Scullion, R.; Harrington, C. S.; Madden, R. H.

2004-01-01

Recent evidence suggests that arcobacters, especially Arcobacter butzleri, are potential foodborne pathogens, but standardized detection methods have yet to be established. A study was undertaken to determine which of three isolation methods was the most effective for the isolation of Arcobacter...... spp. from fresh raw poultry. Method I was microaerobic and involved a membrane filtration step followed by plating onto blood agar. Method 2 was also microaerobic and involved enrichment and plating media containing a five-antibiotic cocktail. Method 3 was aerobic and was based on enrichment...... in a charcoal-based broth containing two antibiotics. Retail poultry samples (n = 50) were obtained from supermarkets in Northern Ireland; the European Community license number was recorded to ensure sample diversity. Presumptive arcobacters were identified using genus-specific and species-specific primers...

Comparison of Methods for Isolating High Quality DNA and RNA from an Oleaginous Fungus Cunninghamella bainieri Strain 2a1

Directory of Open Access Journals (Sweden)

Noor Adila, A. K.

2007-01-01

Full Text Available A number of protocols have been reported for efficient fungal DNA and RNA isolation. However, many of these methods are often designed for certain groups or morphological forms of fungi and, in some cases, are species dependent. In this report, we compared four published protocols for DNA isolation from a locally isolated oleaginous fungus, Cunninghamella bainieri strain 2a1. These protocols either involved the use of polyvinyl pyrrolidone (PVP, hexacetyltrimethylammonium bromide (CTAB or without using PVB or CTAB. For RNA isolation, we tested two published protocols, one of which is based on TRI REAGENT (Molecular Research Center, USA and another is simple method employing phenol for RNA extraction and LiCl for precipitation. We found that the protocol involving the use of CTAB produced the highest genomic DNA yield with the best quality compared to other protocols. In the presence of CTAB, unwanted polysaccharides were removed and this method yielded an average amount of 816 ± 12.2 µg DNA/g mycelia with UV absorbance ratios A260/280 and A260/230 of 1.67 ± 0.64 and 1.97 ± 0.23, respectively. The genomic DNA isolated via this protocol is also suitable for PCR amplification and restriction enzyme digestion. As for RNA isolation, the method involving phenol extraction and LiCl precipitation produced the highest yield of RNA with an average amount of 372 ± 6.0 µg RNA/g mycelia. The RNA appears to be relatively pure since it has UV absorbance ratios A260/280 and A260/230 of 1.89 ± 2.00 and 1.99 ± 0.03, respectively. Finally, we have demonstrated that this method could produce RNA of sufficient quality for RT-PCR that amplified a 600 bp fragment of ∆12-fatty acid desaturase gene in C. bainieri.

In situ macromolecular crystallography using microbeams.

Science.gov (United States)

Axford, Danny; Owen, Robin L; Aishima, Jun; Foadi, James; Morgan, Ann W; Robinson, James I; Nettleship, Joanne E; Owens, Raymond J; Moraes, Isabel; Fry, Elizabeth E; Grimes, Jonathan M; Harlos, Karl; Kotecha, Abhay; Ren, Jingshan; Sutton, Geoff; Walter, Thomas S; Stuart, David I; Evans, Gwyndaf

2012-05-01

Despite significant progress in high-throughput methods in macromolecular crystallography, the production of diffraction-quality crystals remains a major bottleneck. By recording diffraction in situ from crystals in their crystallization plates at room temperature, a number of problems associated with crystal handling and cryoprotection can be side-stepped. Using a dedicated goniometer installed on the microfocus macromolecular crystallography beamline I24 at Diamond Light Source, crystals have been studied in situ with an intense and flexible microfocus beam, allowing weakly diffracting samples to be assessed without a manual crystal-handling step but with good signal to noise, despite the background scatter from the plate. A number of case studies are reported: the structure solution of bovine enterovirus 2, crystallization screening of membrane proteins and complexes, and structure solution from crystallization hits produced via a high-throughput pipeline. These demonstrate the potential for in situ data collection and structure solution with microbeams. © 2012 International Union of Crystallography

Experiments at Scale with In-Situ Visualization Using ParaView/Catalyst in RAGE

Energy Technology Data Exchange (ETDEWEB)

Kares, Robert John [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

2014-10-31

In this paper I describe some numerical experiments performed using the ParaView/Catalyst in-situ visualization infrastructure deployed in the Los Alamos RAGE radiation-hydrodynamics code to produce images from a running large scale 3D ICF simulation on the Cielo supercomputer at Los Alamos. The detailed procedures for the creation of the visualizations using ParaView/Catalyst are discussed and several images sequences from the ICF simulation problem produced with the in-situ method are presented. My impressions and conclusions concerning the use of the in-situ visualization method in RAGE are discussed.

The Role of Isolation Methods on a Nanoscale Surface Structure and Its Effect on the Size of Exosomes

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JungReem Woo

2016-06-01

Full Text Available Exosomes are ~100 nanometre diameter vesicles secreted by mammalian cells. These emerging disease biomarkers carry nucleic acids, proteins and lipids specific to the parental cells that secrete them. Exosomes are typically isolated in bulk by ultracentrifugation, filtration or immu‐ noaffinity precipitation for downstream proteomic, genomic, or lipidomic analysis. However, the structural properties and heterogeneity of isolated exosomes at the single vesicle level are not well characterized due to their small size. In this paper, by using high-resolution atomic force microscope imaging, we show the nanoscale mor‐ phology and structural heterogeneity in exosomes derived from U87 cells. Quantitative assessment of single exosomes reveals nanoscale variations in morphology, surface roughness and counts isolated by ultracentrifugation (UC and immunoaffinity (IA purification. Both methods produce intact globular, 30-120 nm sized vesicles when imaged under fluid and in air. However, IA exosomes had higher surface roughness and bimodal size population compared to UC exosomes. The study highlights the differences in size and surface topography of exosomes purified from a single cell type using different isolation methods.

In situ detrital zircon (U-Th)/He thermochronology

Science.gov (United States)

Tripathy, A.; Monteleone, B. D.; van Soest, M. C.; Hodges, K.; Hourigan, J. K.

2010-12-01

Detrital studies of both sand and rock are relevant to many problems, ranging from the climate and tectonics feedback debate to the long-term record of orogenic evolution. When applying the conventional (U-Th)/He technique to such studies, two important issues arise. Often, only euhedral grains are permissible for analysis in order to make simple geometric corrections for α-recoil. In detrital samples, this is problematic because euhedral grains can be scarce due to mechanical abrasion during transport, and potentially introduce bias in favour of more proximally sourced grains. Second, inherent to detrital studies is the need to date many grains (>100) per sample to ensure a representative sampling of the sediment source region, thus making robust conventional detrital studies both expensive and time-consuming. UV laser microprobes can improve this by permitting careful targeting of the grain interior away from the α-ejection zone, rendering the α-recoil correction unnecessary, thus eliminating bias toward euhedral grains. In the Noble Gas, Geochemistry, and Geochronology Laboratory at ASU, apatite and zircon have been successfully dated using in situ methods. For this study, the conventional and in situ techniques are compared by dating zircons from a modern river sand that drains a small catchment in the Mesozoic-Cenozoic Ladakh Batholith in NW India. This sample has a simple provenance, which allows us to demonstrate the robustness of the in situ method. Moreover, different microbeam techniques will be explored to establish the most efficient approach to obtain accurate and precise U-Th concentrations using synrock, which is our powdered, homogenized, and reconstituted zircon-rock standard. Without this, such in situ U-Th data would be difficult to obtain. 117 zircons were dated using the conventional (U-Th)/He method, revealing dates ranging from 9.70±0.35 to 106.6±3.5 Ma (2σ) with the major mode at 26 Ma. For comparison, 44 grains were dated using the in

Three methods for isolating viable anthozoan endoderm cells with their intracellular symbiotic dinoflagellates

Science.gov (United States)

Gates, R. D.; Muscatine, L.

1992-09-01

Three maceration methods are described for the isolation of single endoderm cells from marine cnidarians. Two are enzymatic treatments suitable for fleshy anthozoans such as sea anemones and zoanthids. The third employs calcium free sea water and is suitable for stony corals. The viability and morphology of the endoderm cells is described using fluorogenic dyes and scanning and transmission electron microscopy.

An accelerated method for the detection of Extended-Spectrum B- Lactamases in urinary isolates of Escherichia Coli and Klebsiella pneumoniae

International Nuclear Information System (INIS)

Kader, Abdulrahman A.; Kumar, A.; Krishna, A.; Zaman, M.N.

2006-01-01

We prospectively studied an accelerated phenotypic method by incorporating the double disk synergy test in the standard Kirby-Bauer disk diffusion susceptibility testing, to evaluate a protocol for the rapid detection of extended of extended-spectrum B-lactamases (ESBL) in urinary isolates of Escherichia coli (E. coli) and Klebsiella, pneumoniae (K. pneumoniae). All ESBL-positive isolates were confirmed by the standard Clinical Laboratory Standards Institute (CLSI) confirmatory disk diffusion method. Between November 2004 and December 2005, a total of 6988 urine specimens were analyzed of which 776 (11%) showed significant growth. They included E. coli in 577 cases (74%) and K. pneumoniae in 199 (25.6%). Of these, 63 E. coli (8%) and 15 K. pneumoniae (7.5%) were positive for ESBL by the accelerated and CLSI methods. Compared to the standard CLSI method, the accelerated method reduced the ESBL detection time from two days to one day. We conclude that the accelerated ESBL detection technique used by us in this study is a reliable and rapid method for detecting ESBL in urinary isolates of E. coli and K. pneumoniae. (author)

Developing biological and chemical methods for environmental monitoring of DOE waste disposal and storage facilities. Progress report, April 1, 1985--October 30, 1985

International Nuclear Information System (INIS)

1997-01-01

During the first year of this contract great efforts were made to develop methods for (1) characterizing bacteria from soil and sediment, (2) evaluating the ability of single and mixed soil bacterial isolates to, (a) bioconcentrate, (b) biodegrade and/or (c) precipitate inorganic and organic pollutants and (3) expanding current concepts for treating waste in aqueous (i.e. biological waste treatment system) and solid media (i.e. in situ soil (soil) treatment system). The development of the above methods are in the final stages of completion and we have as a result of these efforts isolated from soil (1) a mixed culture which precipitate toxic metals (i.e. mercury cadmium, lead etc.) and (2) single isolates which bioconcentrate a variety of toxic metals. Methods for screening soil bacterial isolates for their ability to concentrate, degrade and/or precipitate environmental pollutants have been developed. The development of those methods will allow the staff at ORRI to quickly screen hundreds of samples in our attempt to isolate bacteria capable of degrading, concentrating and/or precipitating inorganics and organics in aqueous and solid waste. The results of these studies are summarized below

Autoradiographic study of transcription during early germination of Zea mays embryos maintained in situ

International Nuclear Information System (INIS)

Deltour, Roger

1979-01-01

Recovery of RNA synthesis was studied by autoradiography in primary root of Zea mays embryos germinating at 16 0 C. [H 3 ] uridine was provided to embryos maintained in situ. During the first 4hrs of germination the cell radioactivity is located almost exclusively in the extranucleolar chromatin. These observations agree well with previous results obtained when [H 3 ] uridine was provided to isolated embryos [fr

Standardizing Umbilical Cord Mesenchymal Stromal Cells for Translation to Clinical Use: Selection of GMP-Compliant Medium and a Simplified Isolation Method

Directory of Open Access Journals (Sweden)

J. Robert Smith

2016-01-01

Full Text Available Umbilical cord derived mesenchymal stromal cells (UC-MSCs are a focus for clinical translation but standardized methods for isolation and expansion are lacking. Previously we published isolation and expansion methods for UC-MSCs which presented challenges when considering good manufacturing practices (GMP for clinical translation. Here, a new and more standardized method for isolation and expansion of UC-MSCs is described. The new method eliminates dissection of blood vessels and uses a closed-vessel dissociation following enzymatic digestion which reduces contamination risk and manipulation time. The new method produced >10 times more cells per cm of UC than our previous method. When biographical variables were compared, more UC-MSCs per gram were isolated after vaginal birth compared to Caesarian-section births, an unexpected result. UC-MSCs were expanded in medium enriched with 2%, 5%, or 10% pooled human platelet lysate (HPL eliminating the xenogeneic serum components. When the HPL concentrations were compared, media supplemented with 10% HPL had the highest growth rate, smallest cells, and the most viable cells at passage. UC-MSCs grown in 10% HPL had surface marker expression typical of MSCs, high colony forming efficiency, and could undergo trilineage differentiation. The new protocol standardizes manufacturing of UC-MSCs and enables clinical translation.

Otoliths in situ from Sarmatian (Middle Miocene) fishes of the Paratethys. Part I

DEFF Research Database (Denmark)

Schwarzhans, Werner; Carnevale, Giorgio; Bannikov, Alexandre F.

2017-01-01

to two otolith-based species so far identified from the same time interval in the Paratethys---Atherina austriaca and Atherina gidjakensis. Our correlation of isolated otoliths and otolith in situ documents in this case that A. suchovi is not synonymous to any of the otolith-based species, although...... it appears to be closely related to A. gidjakensis. A list is presented and briefly discussed showing Sarmatian skeleton-based fish records from the Central and Eastern Paratethys with an overview of known and currently studied fishes with otoliths in situ.......Several well-preserved otoliths were extracted from four slabs containing fish specimens of Atherina suchovi. Atherina suchovi is one of the five Atherina species recorded from the Middle Miocene of the Central and Eastern Paratethys established on articulated skeletal remains. This corresponds...

A method for the isolation of root hairs from the model legume Medicago truncatula

NARCIS (Netherlands)

Ramos Escribano, J.; Bisseling, T.

2003-01-01

A new method for the isolation of root hairs from the model legume, Medicago truncatula, was developed. The procedure involves the propagation of detached roots on agar plates and the collection of root hairs by immersion in liquid nitrogen. Yields of up to 40 µg of root hair protein were obtained

Identification of Candida albicans and Candida dubliniensis Species Isolated from Bronchoalveolar Lavage Samples Using Genotypic and Phenotypic Methods.

Science.gov (United States)

Kianipour, Sahar; Ardestani, Mohammad Emami; Dehghan, Parvin

2018-01-01

Candida dubliniensis is a newly diagnosed species very similar to Candida albicans phenotypically and first discovered in the mouth of people with AIDS in 1995. Among the different phenotypic and genotypic methods, a cost-effective method should be selected which makes it possible to differentiate these similar species. Polymerase chain reaction (PCR)-restriction fragment length polymorphism with MspI enzyme and the Duplex-PCR method were done by DNA extraction using boiling. The sequencing of the amplified ribosomal region was used to confirm the C. dubliniensis species. Direct examination and colony count of the yeasts were applied for bronchoalveolar lavage (BAL) samples and the growth rate of the yeasts were studied at 45°C. To understand the ability formation of chlamydoconidia in yeast isolates, they were separately cultured on the sunflower seed agar, wheat flour agar, and corn meal agar media. Fifty-nine (49.2%) yeast colonies were identified from the total of 120 BAL specimens. Twenty-nine isolated yeasts; including 17 (58.6%) of C. albicans / dubliniensis complex and 12 (41.4%) of nonalbicans isolates produced pseudohypha or blastoconidia in direct smear with a mean colony count of 42000 CFU/mL. C. albicans with the frequency of 15 (42.9%) were the most common isolated yeasts, whereas C. dubliniensis was identified in two nonHIV patients. Sequencing of the replicated gene fragment is the best method for identifying the yeasts, but the determination of the species by phenotypic methods such as the creation of chlamydoconidia in sunflower seeds agar and wheat flour agar media can be cost-effective, have sensitivity and acceptable quality.

Sperm cryopreservation in endangered felids: developing linkage of in situ-ex situ populations.

Science.gov (United States)

Swanson, W F; Magarey, G M; Herrick, J R

2007-01-01

Many of the world's cat species face growing threats to their continued survival in nature. For some species, managed captive populations may provide a reservoir for future reintroduction or genetic augmentation. Because most zoo populations are derived from small founder sizes and are subject to loss of genetic variation over time, periodic infusion of founder alleles is necessary to avoid the dire consequences of inbreeding. Collection and freezing of semen from free-living nondomestic felids offers a viable option for introducing founder genes into captive populations without removal of animals from the wild. The effective application of this strategy requires established protocols for safely capturing and anaesthetising wild cats coupled with suitable methods of semen recovery, processing and cryopreservation under field conditions. In small-sized non-domestic felids, the general feasibility of this approach is being explored in two studies of black-footed cats and Pallas' cats. Two factors - relatively low sperm numbers per ejaculate and compromised status of frozen-thawed cat spermatozoa - suggest that in vitro fertilisation (IVF) and embryo transfer present the most efficient use of this limiting resource in small-sized cats. Our studies with captive felids have explored alternative methods of sperm cryopreservation that are adaptable to field situations and shown that frozen-thawed spermatozoa from Pallas' cats, ocelots, and fishing cats exhibit adequate function to fertilise heterologous and/or homologous oocytes in vitro. Most recently, we investigated the fertilising capacity of frozen-thawed spermatozoa obtained from wild Pallas' cats in Mongolia. Combined with improved methods for embryo culture and transfer in small cat species, sperm banking in situ will facilitate introduction of new founders into captive populations without causing further depletion of their wild counterparts. As one component of holistic conservation programs, including ongoing

Mn{sub 2}O{sub 3}/carbon aerogel microbead composites synthesized by in situ coating method for supercapacitors

Energy Technology Data Exchange (ETDEWEB)

Wang Xingyan, E-mail: wxianyou@yahoo.com [Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, Chemistry School, Xiangtan University, Hunan, Xiangtan 411105 (China); Hunan Institute of Humanities Science and Technology, Loudi 417000 (China); Key Laboratory of Materials Design and Preparation Technology of Hunan, Xiangtan 411105 (China); Liu Li [Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, Chemistry School, Xiangtan University, Hunan, Xiangtan 411105 (China); Wang Xianyou, E-mail: wqinyan801@yahoo.com.cn [Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, Chemistry School, Xiangtan University, Hunan, Xiangtan 411105 (China); Key Laboratory of Materials Design and Preparation Technology of Hunan, Xiangtan 411105 (China); Yi Lanhua [Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, Chemistry School, Xiangtan University, Hunan, Xiangtan 411105 (China); Hu Chuanyue [Hunan Institute of Humanities Science and Technology, Loudi 417000 (China); Zhang Xiaoyan [Key Laboratory of Environmentally Friendly Chemistry and Applications of Ministry of Education, Chemistry School, Xiangtan University, Hunan, Xiangtan 411105 (China)

2011-09-15

Highlights: > Mn{sub 2}O{sub 3}/CAMB composite materials for supercapacitor were prepared by in situ coating method. > The optimum amount of Mn{sub 2}O{sub 3} in Mn{sub 2}O{sub 3}/CAMB composite is 10 wt%. > Coating nano-sized Mn{sub 2}O{sub 3} on the CAMB could improve the supercapacitive behaviors of composites. - Abstract: A series of Mn{sub 2}O{sub 3}/carbon aerogel microbead (Mn{sub 2}O{sub 3}/CAMB) composites for supercapacitor electrodes have been synthesized by in situ encapsulation method. The structure and morphology of Mn{sub 2}O{sub 3}/CAMB are characterized by X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectrum and scanning electron microscopy (SEM). Electrochemical performances of the synthesized composites are evaluated by cyclic voltammetry and galvanostatic charge/discharge measurement. All the composites with different Mn{sub 2}O{sub 3} contents show higher specific capacitance than pure CAMB due to the pseudo-capacitance of the Mn{sub 2}O{sub 3} particles dispersed on the surface of CAMB. The highest specific capacitance is up to 368.01 F g{sup -1} when 10 wt% Mn{sub 2}O{sub 3} is coated on the surface of CAMB. Besides, 10%-Mn{sub 2}O{sub 3}/CAMB supercapacitor exhibits excellent cyclic stability, the specific capacitance still retains 90% of initial capacitance over 5000 cycles.

Superior electrode performance of LiFePO4/C composite prepared by an in situ polymerization restriction method

International Nuclear Information System (INIS)

Chen, Jian; Zou, Yong-Cun; Zhang, Feng; Zhang, Yuan-Chun; Guo, Fei-Fan; Li, Guo-Dong

2013-01-01

Highlights: ► LiFePO 4 /C composite was prepared by an in situ polymerization restriction method. ► The size of LiFePO 4 in the composite is effectively restricted. ► The high-rate capability and cycling performance of LiFePO 4 are enhanced greatly. -- Abstract: The LiFePO 4 /C composite is prepared by heating the mixture of resorcinol–formaldehyde gel and FePO 4 , synthesized by an in situ polymerization restriction method, and lithium acetate dihydrate in the atmosphere of nitrogen. The physical and electrochemical properties of the LiFePO 4 /C composite are investigated by X-ray diffraction, Raman spectroscopy, thermogravimetric analysis, scanning electron microscopy, transmission electron microscopy and electrochemical measurements. The discharge capacity of LiFePO 4 is as high as 155.6 mA h g −1 in the first cycle at 0.5C, and it could remain 144.0 mA h g −1 after 50 cycles. Even at the high rates of 10C, 20C and 50C, the initial discharge capacities of the electrodes exhibit 115.6 mA h g −1 , 84.5 mA h g −1 and 67.8 mA h g −1 , and the electrodes deliver capacity retention of 89.5%, 90.9% and 85.7% after 1000 cycles, respectively. The outstanding electrochemical performance could be attributed to the small particle size and good electronic conductivity of the composite

In-situ observation of structure formation in polymer processing

International Nuclear Information System (INIS)

Murase, Hiroki

2009-01-01

In-situ X-ray scattering in polymer processing is a crucial method to elucidate the mechanism of structure formation in the process. Fiber spinning is one such process primarily imposing extensional deformation on polymeric melt at the spin-line during rapid cooling. In-situ small-angle X-ray scattering using synchrotron radiation on the spinning process allows direct observation of the transient structure developing in the process. (author)

The isolation of nucleic acids from fixed, paraffin-embedded tissues-which methods are useful when?

DEFF Research Database (Denmark)

Gilbert, M Thomas P; Haselkorn, Tamara; Bunce, Michael

2007-01-01

. Cross-linking not only complicates isolation of nucleic acid but also introduces polymerase "blocks" during PCR. A wide variety of methods exists for the recovery of DNA and RNA from archival tissues, and although a number of previous studies have qualitatively compared the relative merits....... These include methods of pre-treating the samples prior to extraction, extraction and nucleic acid purification methods themselves, and a post-extraction enzymatic repair technique. We find that although many of the published methods have distinct positive effects on some characteristics of the nucleic acids...

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

Science.gov (United States)

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

2017-11-01

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

Effects of Phytophthora cinnamomi isolate, inoculum delivery method, flood, and drought on vigor, disease severity and mortality of blueberry plants

Science.gov (United States)

Four studies evaluated the effect of Phytophthora cinnamomi isolates, inoculum delivery methods, and flood and drought conditions on vigor, disease severity scores, and survival of blueberry plants grown in pots in the greenhouse. Phytophthora cinnamomi isolates were obtained from blueberry plants ...

New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

Directory of Open Access Journals (Sweden)

Karen Nava-Castro

2011-01-01

Full Text Available In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells.

Influence of Steel Reinforcement on In-Situ Stress Evaluation in Concrete Structures by the Core-Drilling Method

International Nuclear Information System (INIS)

McGinnis, M. J.; Pessiki, S.

2006-01-01

The core-drilling method is an emerging technique for evaluating in-situ stress in a concrete structure. A small hole is drilled into the structure, and the deformations in the vicinity of the hole are measured and related via elasticity theory to the stress. The method is similar to the ASTM hole-drilling strain-gauge method excepting that displacements rather than strains are the measured quantities. The technique may be considered nondestructive since the ability of the structure to perform its function is unaffected, and the hole is easily repaired. Displacement measurements in the current work are performed using 3D digital image correlation and industrial photogrammetry. The current paper addresses perturbations in the method caused by steel reinforcement within the concrete. The reinforcement is significantly stiffer than the surrounding concrete, altering the expected displacement field. A numerical investigation performed indicates an under-prediction of stress by as much as 18 percent in a heavily reinforced structure, although the effect is significantly smaller for more common amounts of reinforcement

Influence of Steel Reinforcement on In-Situ Stress Evaluation in Concrete Structures by the Core-Drilling Method

Science.gov (United States)

McGinnis, M. J.; Pessiki, S.

2006-03-01

The core-drilling method is an emerging technique for evaluating in-situ stress in a concrete structure. A small hole is drilled into the structure, and the deformations in the vicinity of the hole are measured and related via elasticity theory to the stress. The method is similar to the ASTM hole-drilling strain-gauge method excepting that displacements rather than strains are the measured quantities. The technique may be considered nondestructive since the ability of the structure to perform its function is unaffected, and the hole is easily repaired. Displacement measurements in the current work are performed using 3D digital image correlation and industrial photogrammetry. The current paper addresses perturbations in the method caused by steel reinforcement within the concrete. The reinforcement is significantly stiffer than the surrounding concrete, altering the expected displacement field. A numerical investigation performed indicates an under-prediction of stress by as much as 18 percent in a heavily reinforced structure, although the effect is significantly smaller for more common amounts of reinforcement.

Ambient ex-situ denitrification in isolated wetlands of Ohio, North Carolina and Florida

Science.gov (United States)

Isolated wetlands are completely surrounded by uplands and typically do not warrant federal protection under the Clean Water Act. Nevertheless they can be found at high densities in certain parts of the US and Canada (e.g., Prairie Pothole Region, Southern and Middle Atlantic Coa...

Engineering feasibility analysis for in-situ stabilization of Canonsburg residues

International Nuclear Information System (INIS)

1982-01-01

The US Department of Energy is considering several methods for carrying out remedial actions in Canonsburg, Pennsylvania, at the site of an inactive uranium-processing mill. The main objective of this study is to determine the feasibility of in-situ stabilization as the remedial action. In-situ stabilization is an alternative to site decontamination and offsite disposal. The problems associated with offsite hauling of large quantities of contaminated material and with the location and development of a new disposal site could be avoided by the implementation of an in-situ stabilization concept. In addition, the in-situ approach would be more cost-effective than offsite disposal. This study will establish that a technically feasible and implementable in-situ stabilization concept can be developed that meets regulatory requirements and is cost effective. This study in no way commits the DOE to implement any specific actions described herein. 11 refs., 30 figs., 24 tabs

In situ hybridization; principles and applications: review article

Directory of Open Access Journals (Sweden)

Zahra Nozhat

2015-06-01

Full Text Available In situ hybridization (ISH is a method that uses labeled complementary single strand DNA or RNA to localize specific DNA or RNA sequences in an intact cell or in a fixed tissue section. The main steps of ISH consist of: probe selection, tissue or sample preparation, pre-hybridization treatment, hybridization and washing, detection and control procedure. Probe selection is one of the important aspects of successful hybridization. ISH sensitivity and specificity can be influenced by: probe construct, efficiency of labeling, percentage of GC, probe length and signal detection systems. Different methods such as nick translation, random priming, end tailing and T4 DNA polymerase replacement are used for probe generation. Both radioactive and non-radioactive labels can be used in order to probe labeling. Nucleic acid maintenance in samples, prevention of morphological changes of samples and probe penetration into tissue section are the main aims of sample preparation step. Then, a small amount of solution containing probe, is added on slides containing tissue sections for hybridization process, then slides are incubated overnight. Next day, washes are carried out to remove the probes which are not bound to target DNA or RNA. Finally, in order to be sure that the observed labeling is specific to the target sequence, using several control procedures is very important. Various techniques based on ISH consist of: Fluorescence in situ hybridization (FISH, chromogenic in situ hybridization (CISH, genomic in situ hybridization (GISH, comparative genomic hybridization (CGH, spectral karyotyping (SKY and multiplex fluorescence in situ hybridization (MFISH. One of the most common techniques of ISH is fluorescence in situ hybridization. FISH can be used to: 1 detect small deletions and duplications that are not visible using microscope analysis, 2 detect how many chromosomes of a certain type are present in each cell and 3 confirm rearrangements that are

Development of an in-situ banking shear test method; Moritsuchi no gen'ichi sendan shikenho no kaihatsu

Energy Technology Data Exchange (ETDEWEB)

Nishikata, U. [Kansai Electric Power Co. Inc., Osaka (Japan)

1998-12-10

Development was made on a simplified site-use single face shear testing device and a test method that can perform in-situ and direct measurement during construction on shear strength of coarse particle materials having large diameters. The test device consists of shear frames embedded in the in-situ ground bed, a weight for vertical loading, and a traction device to load horizontal force. In the test, prescribed rolling compaction is performed by a vibration roller which presses in the shear frames into unwound banking by using a four-piece vibration roller. Crushed stones are piled on cells of the shear frames to the same height. The specified weight is placed thereon via a loading plate, which is used as the vertical load. The shear force is loaded by horizontally pulling chains connected to the shear frames by using the traction device, and is measured by a load cell. The vertical displacement is measured by two vertical displacement meters, and the horizontal displacement by one horizontal displacement meter. A verification test is continuing using the test device. Four cases with different vertical stresses can be tested in about one day. (NEDO)

The application of geophysical logging at in-situ leaching uranium mine in China

International Nuclear Information System (INIS)

Liu Zeyao; Xu Shusheng; Li Zhongqiu

1999-01-01

The status of work, instrument and method employed for geophysical logging in different stages at in-situ leaching uranium mine are discussed and the development of software, electrical current logging and gamma ray logging are presented based on the requirement of in-situ leaching of uranium. In addition, new function and method with regard to home instrument are proposed for future work

Comparison of in-situ gamma ray spectrometry measurements with conventional methods in determination natural and artificial nuclides in soil

International Nuclear Information System (INIS)

Al-Masri, M. S.; Doubal, A. W.

2010-12-01

Two nuclear analytical techniques (In-Situ Gamma ray spectrometry and laboratory gamma ray spectrometry) for determination of natural and artificial radionuclides in soil have been validated. The first technique depends on determination of radioactivity content of representative samples of the studied soil after laboratory preparation, while the second technique is based on direct determination of radioactivity content of soil using in-situ gamma-ray spectrometer. Analytical validation parameter such as detection limits, repeatability, reproducibility in addition to measurement uncertainties were estimated and compared for both techniques. Comparison results have shown that the determination of radioactivity in soil should apply the two techniques together where each of techniques is characterized by its low detection limit and uncertainty suitable for defined application of measurement. Radioactive isotopes in various locations were determined using the two methods by measuring 40 k, 238 U,and 137 Cs. The results showed that there are differences in attenuation factors due to soil moisture content differences; wet weight corrections should be applied when the two techniques are compared. (author)

In-situ measurements of the secondary electron yield in an accelerator environment: Instrumentation and methods

International Nuclear Information System (INIS)

Hartung, W.H.; Asner, D.M.; Conway, J.V.; Dennett, C.A.; Greenwald, S.; Kim, J.-S.; Li, Y.; Moore, T.P.; Omanovic, V.; Palmer, M.A.; Strohman, C.R.

2015-01-01

The performance of a particle accelerator can be limited by the build-up of an electron cloud (EC) in the vacuum chamber. Secondary electron emission from the chamber walls can contribute to EC growth. An apparatus for in-situ measurements of the secondary electron yield (SEY) in the Cornell Electron Storage Ring (CESR) was developed in connection with EC studies for the CESR Test Accelerator program. The CESR in-situ system, in operation since 2010, allows for SEY measurements as a function of incident electron energy and angle on samples that are exposed to the accelerator environment, typically 5.3 GeV counter-rotating beams of electrons and positrons. The system was designed for periodic measurements to observe beam conditioning of the SEY with discrimination between exposure to direct photons from synchrotron radiation versus scattered photons and cloud electrons. The samples can be exchanged without venting the CESR vacuum chamber. Measurements have been done on metal surfaces and EC-mitigation coatings. The in-situ SEY apparatus and improvements to the measurement tools and techniques are described

In-situ measurements of the secondary electron yield in an accelerator environment: Instrumentation and methods

Energy Technology Data Exchange (ETDEWEB)

Hartung, W.H., E-mail: wh29@cornell.edu; Asner, D.M.; Conway, J.V.; Dennett, C.A.; Greenwald, S.; Kim, J.-S.; Li, Y.; Moore, T.P.; Omanovic, V.; Palmer, M.A.; Strohman, C.R.

2015-05-21

The performance of a particle accelerator can be limited by the build-up of an electron cloud (EC) in the vacuum chamber. Secondary electron emission from the chamber walls can contribute to EC growth. An apparatus for in-situ measurements of the secondary electron yield (SEY) in the Cornell Electron Storage Ring (CESR) was developed in connection with EC studies for the CESR Test Accelerator program. The CESR in-situ system, in operation since 2010, allows for SEY measurements as a function of incident electron energy and angle on samples that are exposed to the accelerator environment, typically 5.3 GeV counter-rotating beams of electrons and positrons. The system was designed for periodic measurements to observe beam conditioning of the SEY with discrimination between exposure to direct photons from synchrotron radiation versus scattered photons and cloud electrons. The samples can be exchanged without venting the CESR vacuum chamber. Measurements have been done on metal surfaces and EC-mitigation coatings. The in-situ SEY apparatus and improvements to the measurement tools and techniques are described.

Localization of tRNAsup(asp)2 genes from Drosophila melanogaster by 'in situ' hybridization

International Nuclear Information System (INIS)

Schmidt, T.; Egg, A.H.; Kubli, E.

1978-01-01

Transfer RNAsup(asp) 2 delta was isolated from Drosophila melanogaster by affinity chromatography on concanavalin A-Sepharose. The tRNA was iodinated 'in vitro' with Na[ 125 I] and hybridized 'in situ' to salivary gland chromosomes from Drosophila. Subsequent autoradiography allowed the localization of the genes for tRNAsup(asp) 2 delta to the left arm of the second chromosome in the regions 29 D and E. (orig.) [de

Styrene grafted natural rubber reinforced by in situ silica generated via sol–gel technique

Energy Technology Data Exchange (ETDEWEB)

Sittiphan, Torpong [Program of Petrochemistry and Polymer Sciences, Faculty of Science, Chulalongkorn University, Bangkok 10330 (Thailand); Prasassarakich, Pattarapan [Department of Chemical Technology, Faculty of Science, Chulalongkorn University, Bangkok 10330 (Thailand); Poompradub, Sirilux, E-mail: sirilux.p@chula.ac.th [Department of Chemical Technology, Faculty of Science, Chulalongkorn University, Bangkok 10330 (Thailand)

2014-02-15

Highlights: • Sol–gel reaction by NR latex was the absence of use of organic solvent and base catalyst. • Well dispersed in situ formed silica particles in the rubber matrix were obtained. • In situ silica was better to improve mechanical properties of rubber vulcanizates. -- Abstract: The filling of styrene graft natural rubber (ST-GNR) with in situ formed silica was performed using the sol–gel reaction via the latex solution method. The mechanical properties of ST-GNR/NR vulcanizate were improved when using the in situ formed silica to levels higher than those obtained with the commercial ex situ formed silica filled ST-GNR/NR vulcanizates at a comparable silica content of 12 parts by weight per hundred parts of rubber. Transmission electron microscopy analysis revealed that the in situ silica particles were small (∼40 nm diameter) and well dispersed, while the commercial silica particles were larger (∼60 nm diameter) and markedly agglomerated in the rubbery matrix. The mechanical properties of the composites prepared via both the solid rubber and latex solution methods were comparable.

Effect of In-Situ Curing on Compressive Strength of Reactive Powder Concrete

Directory of Open Access Journals (Sweden)

Bali Ika

2016-01-01

Full Text Available A development of Reactive Powder Concrete (RPC currently is the use of quartz powder as a stabilizing agent with the content to cement ratio of 30% and steam curing method in an autoclave temperature of 250ºC which produced a high compressive strength of 180 MPa. That RPC can be generated due to one reason for using the technique of steam curing in an autoclave in the laboratory. This study proposes in-situ curing method in order the curing can be applied in the field and with a reasonable compressive strength results of RPC. As the benchmarks in this study are the curing methods in laboratory that are steam curing of 90°C for 8 hours (C1, and water curing for 28 days (C2. For the in-situ curing methods that are covering with tarpaulins and flowed steam of 3 hours per day for 7 days (C3, covering with wet sacks for 28 days (C4, and covering with wet sacks for 28 days for specimen with unwashed sand as fine aggregate (C5. The comparison of compressive strength of the specimens in this study showed compressive strength of RPC with in-situ steam curing (101.64 MPa close to the compressive strength of RPC with steam curing in the laboratory with 8.2% of different. While in-situ wet curing compared with the water curing in laboratory has the different of 3.4%. These results indicated that the proposed in-situ curing methods are reasonable good in term of the compressive strength that can be achieved.

In Situ Magnetic Separation for Extracellular Protein Production

DEFF Research Database (Denmark)

Kappler, T.; Cerff, Martin; Ottow, Kim Ekelund

2009-01-01

A new approach for in situ product removal from bioreactors is presented in which high-gradient magnetic separation is used. This separation process was used for the adsorptive removal of proteases secreted by Bacillus licheniformis. Small, non-porous bacitracin linked magnetic adsorbents were...... was not influenced by the in situ product removal step. Protease production also remained the same after the separation step. Furthermore, degradation of the protease, which followed first order kinetics, was reduced by using the method. Using a theoretical modeling approach, we Could show that protease yield...... in total was enhanced by using in situ magnetic separation. The process described here is a promising technique to improve overall yield in No production processes which are often limited due to weak downstream operations, Potential limitations encountered during a bioprocess can be overcome...

PENGARUH KUALITAS PELAYANAN TERHADAP KEPUASAN BERKUNJUNG DI TAMAN OBJEK WISATA SITU PATENGGANG

Directory of Open Access Journals (Sweden)

Siti Fatimah

2016-12-01

ABSTRACT Situ Patenggang or Situ Patengan is a lake located in South Bandung nature tourist attraction, Jawa Barat, Indonesia, Ciwidey, to be exact. Located 1600 metres above see level, this lake has a really exotic view. Situ Patenggang has a beautiful natural view as well, because of the tea garden surrounding it. The managers should give their best service to make the customers satisfied. This research’s population is the visitor of Situ Patenggang Tourist Attraction Park, with a sample of 30 respondents and the technique used was nonprobability sampling. Data collection method was by using questionnaire given to 100 visitors of Situ Patenggang Tourist Attraction Park and documentation method. The result shows that service quality affects the satisfactory of 47.9% visitors of Situ Patenggang Tourist Attraction Park while the rest, which is 52.1%, was affected by other variables which was not investigated in this research. According to the result of the research done in Situ Patenggang Tourist Attraction Park, it can be seen from the score and mean categorization in the research result’s statistics data regarding the service quality of Situ Patenggang Tourist Attraction Park can be categorized as well enough, with the percentage of 52.1% at the score range of 4069. Whereas according to the score and mean categorization in the research result’s statistics data regarding the customer satisfactory of Situ Patenggang Tourist Attraction Park can be categorized as not good, with the percentage of 52% at the score range of 2080. Therefore, the service quality really takes effect to the satisfaction in visiting, if the service quality is good then the customers will be pleasant and satisfied of their visit.   Keywords: Service Quality, Satisfaction, Tourist Attraction.

In situ macromolecular crystallography using microbeams

International Nuclear Information System (INIS)

Axford, Danny; Owen, Robin L.; Aishima, Jun; Foadi, James; Morgan, Ann W.; Robinson, James I.; Nettleship, Joanne E.; Owens, Raymond J.; Moraes, Isabel; Fry, Elizabeth E.; Grimes, Jonathan M.; Harlos, Karl; Kotecha, Abhay; Ren, Jingshan; Sutton, Geoff; Walter, Thomas S.; Stuart, David I.; Evans, Gwyndaf

2012-01-01

A sample environment for mounting crystallization trays has been developed on the microfocus beamline I24 at Diamond Light Source. The technical developments and several case studies are described. Despite significant progress in high-throughput methods in macromolecular crystallography, the production of diffraction-quality crystals remains a major bottleneck. By recording diffraction in situ from crystals in their crystallization plates at room temperature, a number of problems associated with crystal handling and cryoprotection can be side-stepped. Using a dedicated goniometer installed on the microfocus macromolecular crystallography beamline I24 at Diamond Light Source, crystals have been studied in situ with an intense and flexible microfocus beam, allowing weakly diffracting samples to be assessed without a manual crystal-handling step but with good signal to noise, despite the background scatter from the plate. A number of case studies are reported: the structure solution of bovine enterovirus 2, crystallization screening of membrane proteins and complexes, and structure solution from crystallization hits produced via a high-throughput pipeline. These demonstrate the potential for in situ data collection and structure solution with microbeams

In situ macromolecular crystallography using microbeams

Energy Technology Data Exchange (ETDEWEB)

Axford, Danny; Owen, Robin L.; Aishima, Jun [Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom); Foadi, James [Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom); Imperial College, London SW7 2AZ (United Kingdom); Morgan, Ann W.; Robinson, James I. [University of Leeds, Leeds LS9 7FT (United Kingdom); Nettleship, Joanne E.; Owens, Raymond J. [Research Complex at Harwell, Rutherford Appleton Laboratory R92, Didcot, Oxfordshire OX11 0DE (United Kingdom); Moraes, Isabel [Imperial College, London SW7 2AZ (United Kingdom); Fry, Elizabeth E.; Grimes, Jonathan M.; Harlos, Karl; Kotecha, Abhay; Ren, Jingshan; Sutton, Geoff; Walter, Thomas S. [University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Stuart, David I. [Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom); University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Evans, Gwyndaf, E-mail: gwyndaf.evans@diamond.ac.uk [Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom)

2012-04-17

A sample environment for mounting crystallization trays has been developed on the microfocus beamline I24 at Diamond Light Source. The technical developments and several case studies are described. Despite significant progress in high-throughput methods in macromolecular crystallography, the production of diffraction-quality crystals remains a major bottleneck. By recording diffraction in situ from crystals in their crystallization plates at room temperature, a number of problems associated with crystal handling and cryoprotection can be side-stepped. Using a dedicated goniometer installed on the microfocus macromolecular crystallography beamline I24 at Diamond Light Source, crystals have been studied in situ with an intense and flexible microfocus beam, allowing weakly diffracting samples to be assessed without a manual crystal-handling step but with good signal to noise, despite the background scatter from the plate. A number of case studies are reported: the structure solution of bovine enterovirus 2, crystallization screening of membrane proteins and complexes, and structure solution from crystallization hits produced via a high-throughput pipeline. These demonstrate the potential for in situ data collection and structure solution with microbeams.

Static and dynamic stability of pneumatic vibration isolators and systems of isolators

Science.gov (United States)

Ryaboy, Vyacheslav M.

2014-01-01

Pneumatic vibration isolation is the most widespread effective method for creating vibration-free environments that are vital for precise experiments and manufacturing operations in optoelectronics, life sciences, microelectronics, nanotechnology and other areas. The modeling and design principles of a dual-chamber pneumatic vibration isolator, basically established a few decades ago, continue to attract attention of researchers. On the other hand, behavior of systems of such isolators was never explained in the literature in sufficient detail. This paper covers a range of questions essential for understanding the mechanics of pneumatic isolation systems from both design and application perspectives. The theory and a model of a single standalone isolator are presented in concise form necessary for subsequent analysis. Then the dynamics of a system of isolators supporting a payload is considered with main attention directed to two aspects of their behavior: first, the static stability of payloads with high positions of the center of gravity; second, dynamic stability of the feedback system formed by mechanical leveling valves. The direct method of calculating the maximum stable position of the center of gravity is presented and illustrated by three-dimensional stability domains; analytic formulas are given that delineate these domains. A numerical method for feedback stability analysis of self-leveling valve systems is given, and the results are compared with the analytical estimates for a single isolator. The relation between the static and dynamic phenomena is discussed.

Calibration of an experimental model of tritium storage bed designed for 'in situ' accountability

International Nuclear Information System (INIS)

Bidica, Nicolae; Stefanescu, Ioan; Bucur, Ciprian; Bulubasa, Gheorghe; Deaconu, Mariea

2009-01-01

Full text: Objectives: Tritium accountancy of the storage beds in tritium facilities is an important issue for tritium inventory control. The purpose of our work was to perform calibration of an experimental model of tritium storage bed with a special design, using electric heaters to simulate tritium decay, and to evaluate the detection limit of the accountancy method. The objective of this paper is to present an experimental method used for calibration of the storage bed and the experimental results consisting of calibration curves and detection limit. Our method is based on a 'self-assaying' tritium storage bed. The basic characteristics of the design of our storage bed consists, in principle, of a uniform distribution of the storage material on several copper thin fins (in order to obtain a uniform temperature field inside the bed), an electrical heat source to simulate the tritium decay heat, a system of thermocouples for measuring the temperature field inside the bed, and good thermal isolation of the bed from the external environment. Within this design of the tritium storage bed, the tritium accounting method is based on determining the decay heat of tritium by measuring the temperature increase of the isolated storage bed. Experimental procedure consisted in measuring of temperature field inside the bed for few values of the power injected with the aid of electrical heat source. Data have been collected for few hours and the temperature increase rate was determined for each value of the power injected. Graphical representation of temperature rise versus injected powers was obtained. This accounting method of tritium inventory stored as metal tritide is a reliable solution for in-situ tritium accountability in a tritium handling facility. Several improvements can be done regarding the design of the storage bed in order to improve the measurement accuracy and to obtain a lower detection limit as for instance use of more accurate thermocouples or special

In situ shape and distance measurements in neutron scattering and diffraction

International Nuclear Information System (INIS)

Fujiwara, Satoru; Mendelson, R.A.

1994-01-01

Neutron scattering combined with selective isotopic labeling and contrast matching is useful for obtaining in situ structural information about a selected particle, or particles, in a macromolecular complex. The observed intensities, however, may be distorted by inter-complex interference and by scattering-length-density fluctuations of the (otherwise) contrast-matched portions. Methods have been proposed to cancel out such distortions (Hoppe's method, the Statistical Labeling Method, and the Triple Isotopic Substitution Method). With these methods as well as related unmixed-sample methods, structural information about the selected particles can be obtained without these distortions. We have generalized these methods so that, in addition to globular particles in solution, they can be applied to in situ structures of systems having underlying symmetry and/or net orientation as well. The information obtainable from such experiments is discussed

Summary of the systems prioritization method as a decision-aiding method for the waste isolation pilot plant

International Nuclear Information System (INIS)

Boak, D.M.; Prindle, N.H.; Lincoln, R.

1996-12-01

In March 1994, the U.S. Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance-based decision-aiding method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) Project with respect to applicable U.S. Environmental Protection Agency (EPA) long-term performance requirements in 40 CFR 191.13(a) (radionuclide containment requirements) and 40 CFR 268.6 (hazardous constituent concentration requirements). This method, the Systems Prioritization Method (SPM), was designed by Sandia National Laboratories (SNL) to: (1) identify programmatic options (activities) and their costs and durations; (2) analyze combinations of activities (activity sets) in terms of their predicted contribution to long-term performance of the WIPP disposal system; and (3) analyze cost, duration, and performance tradeoffs. The results of the second iteration of SPM (SPM-2) were the basis for recommendations to DOE/CAO in May 1995 for programmatic prioritization within the WIPP project. This paper presents a summary of the SPM implementation, key results, and lessons learned

Summary of the systems prioritization method as a decision-aiding method for the waste isolation pilot plant

Energy Technology Data Exchange (ETDEWEB)

Boak, D.M.; Prindle, N.H.; Lincoln, R. [and others

1996-12-01

In March 1994, the U.S. Department of Energy Carlsbad Area Office (DOE/CAO) implemented a performance-based decision-aiding method to assist in programmatic prioritization within the Waste Isolation Pilot Plant (WIPP) Project with respect to applicable U.S. Environmental Protection Agency (EPA) long-term performance requirements in 40 CFR 191.13(a) (radionuclide containment requirements) and 40 CFR 268.6 (hazardous constituent concentration requirements). This method, the Systems Prioritization Method (SPM), was designed by Sandia National Laboratories (SNL) to: (1) identify programmatic options (activities) and their costs and durations; (2) analyze combinations of activities (activity sets) in terms of their predicted contribution to long-term performance of the WIPP disposal system; and (3) analyze cost, duration, and performance tradeoffs. The results of the second iteration of SPM (SPM-2) were the basis for recommendations to DOE/CAO in May 1995 for programmatic prioritization within the WIPP project. This paper presents a summary of the SPM implementation, key results, and lessons learned.

A new method for the determination of radionuclide distribution in the soil by in situ gamma-ray spectrometry

International Nuclear Information System (INIS)

Zombori, P.; Andrasi, A.; Nemeth, I.

1995-01-01

In case of major nuclear accidents when larger amount of radioactive material is released into the atmosphere vast areas can become contaminated by the nuclear fallout. The deposited radioactivity penetrates the soil in a complex manner: dry and wet deposition lead to different initial distribution patterns which are further modified by the later transport processes in the upper layers of the soil. The distribution is influenced by various factors (physico-chemical characteristics of the radioisotopes, soil type, weather conditions, environment etc.), the resulting soil profile is hardly predictable. An important lesson we learned from the Chernobyl reactor accident is the great variability of the contamination both in the extent of the deposition and in the penetration features. In recent years - following the reactor accident in Chernobyl - an increased interest for rapid methods of monitoring environmental radioactivity was expressed. The International Atomic Energy Agency initiated a research project to co-ordinate the activities carried out in various laboratories aiming at the development of rapid monitoring procedures. The Co-ordinated Research Project (CRP) G6 10 01 under the title Rapid Instrumental and Separation Methods for Monitoring Radionuclides in Food and the Environment has given a frame for 11 research programs. The Health Physics Department of the KFKI Institute for Atomic Energy Research (the former Central Research Institute for Physics) has taken a part in this CRP with a project titled: Rapid In Situ Gamma Spectrometric Determination of Fallout Radioactivity in the Environment. The main objective of our study was to find a method to estimate the penetration characteristics of the fallout radioactivity by using only spectral information obtained by the in situ spectrometric measurement thus avoiding the need for a long and tiresome sampling and sample analysis procedure

Comparison of Methods for Isolating High Quality DNA and RNA from an Oleaginous Fungus Cunninghamella bainieri Strain 2a1

OpenAIRE

Noor Adila, A. K.; Farah Diba, A. B.; Zamri, Z.; Wan Mohtar, W. Y.; Aidil, A. H.; Mahadi, N. M.; Murad, A. M. A.

2007-01-01

A number of protocols have been reported for efficient fungal DNA and RNA isolation. However, many of these methods are often designed for certain groups or morphological forms of fungi and, in some cases, are species dependent. In this report, we compared four published protocols for DNA isolation from a locally isolated oleaginous fungus, Cunninghamella bainieri strain 2a1. These protocols either involved the use of polyvinyl pyrrolidone (PVP), hexacetyltrimethylammonium bromide (CTAB) or w...

High Performance Harmonic Isolation By Means of The Single-phase Series Active Filter Employing The Waveform Reconstruction Method

DEFF Research Database (Denmark)

Senturk, Osman Selcuk; Hava, Ahmet M.

2009-01-01

current sampling delay reduction method (SDRM), a single-phase SAF compensated system provides higher harmonic isolation performance and higher stability margins compared to the system using conventional synchronous reference frame based methods. The analytical, simulation, and experimental studies of a 2...

Application of in situ measurement for site remediation and final status survey of decommissioning KRR site

Energy Technology Data Exchange (ETDEWEB)

Hong, Sang Bum; Nam, Jong Soo; Choi, Yong Suk; Seo, Bum Kyoung; Moon, Jei Kwon [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2016-06-15

In situ gamma spectrometry has been used to measure environmental radiation, assumptions are usually made about the depth distribution of the radionuclides of interest in the soil. The main limitation of in situ gamma spectrometry lies in determining the depth distribution of radionuclides. The objective of this study is to develop a method for subsurface characterization by in situ measurement. The peak to valley method based on the ratio of counting rate between the photoelectric peak and Compton region was applied to identify the depth distribution. The peak to valley method could be applied to establish the relation between the spectrally derived coefficients (Q) with relaxation mass per unit area (β) for various depth distribution in soil. The in situ measurement results were verified by MCNP simulation and calculated correlation equation. In order to compare the depth distributions and contamination levels in decommissioning KRR site, in situ measurement and sampling results were compared. The in situ measurement results and MCNP simulation results show a good correlation for laboratory measurement. The simulation relationship between Q and source burial for the source layers have exponential relationship for a variety depth distributions. We applied the peak to valley method to contaminated decommissioning KRR site to determine a depth distribution and initial activity without sampling. The observed results has a good correlation, relative error between in situ measurement with sampling result is around 7% for depth distribution and 4% for initial activity. In this study, the vertical activity distribution and initial activity of {sup 137}Cs could be identifying directly through in situ measurement. Therefore, the peak to valley method demonstrated good potential for assessment of the residual radioactivity for site remediation in decommissioning and contaminated site.

Development and Validation of a SPME-GC-MS Method for In situ Passive Sampling of Root Volatiles from Glasshouse-Grown Broccoli Plants Undergoing Below-Ground Herbivory by Larvae of Cabbage Root Fly, Delia radicum L.

Science.gov (United States)

Deasy, William; Shepherd, Tom; Alexander, Colin J; Birch, A Nicholas E; Evans, K Andrew

2016-11-01

Research on plant root chemical ecology has benefited greatly from recent developments in analytical chemistry. Numerous reports document techniques for sampling root volatiles, although only a limited number describe in situ collection. To demonstrate a new method for non-invasive in situ passive sampling using solid phase micro extraction (SPME), from the immediate vicinity of growing roots. SPME fibres inserted into polyfluorotetrafluoroethylene (PTFE) sampling tubes located in situ which were either perforated, covered with stainless steel mesh or with microporous PTFE tubing, were used for non-invasive sub-surface sampling of root volatiles from glasshouse-grown broccoli. Sampling methods were compared with above surface headspace collection using Tenax TA. The roots were either mechanically damaged or infested with Delia radicum larvae. Principal component analysis (PCA) was used to investigate the effect of damage on the composition of volatiles released by broccoli roots. Analyses by gas chromatography-mass spectrometry (GC-MS) with SPME and automated thermal desorption (ATD) confirmed that sulphur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots following insect larval damage. Use of SPME with in situ perforated PTFE sampling tubes was the most robust method for out-of-lab sampling. This study describes a new method for non-invasive passive sampling of volatiles in situ from intact and insect damaged roots using SPME. The method is highly suitable for remote sampling and has potential for wide application in chemical ecology/root/soil research. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Fluorescence in situ hybridization on formalin-fixed and paraffin-embedded tissue

DEFF Research Database (Denmark)

Laub Petersen, Bodil; Zeuthen, Mette Christa; Pedersen, Sanni

2004-01-01

Fluorescence in situ hybridization (FISH) is widely used to study numerical and structural genetic abnormalities in both metaphase and interphase cells. The technique is based on the hybridization of labeled probes to complementary sequences in the DNA or RNA of the cells. Interphase FISH is most...... in time lapse between removal of tissue and fixation, duration of fixation, enzymatic pretreatment, hybridization conditions, and posthybridization washing conditions are important factors in the hybridization. In this study, we have listed the results of a systematic approach to improve FISH on isolated...

Comparative Analysis of the Genomic DNA Isolation Methods on Inula sp. (Asteraceae

Directory of Open Access Journals (Sweden)

Emre SEVİNDİK

2016-12-01

Full Text Available Simple, fast, low-cost and high throughput protocols are required for DNA isolation of plant species. In this study, phenol chloroform isoamyl alcohol and commercial (Sigma DNA isolation kit methods were applied on some Inula species that belong to Asteraceae family. Genomic DNA amounts, A260, A280, A260/A230 and purity degrees (A260/A280 that were obtained through both methods were measured through electrophoresis and spectrophotometer. Additionally, PCR amplification was realized by primer pairs specific to nrDNA ITS, cpDNA ndhF (972F-1603R and trnL-F regions. Results showed that maximum genomic DNA in nanograms obtained by phenol chloroform isoamyl alcohol method. The study also revealed that I. macrocephala had the maximum DNA and I. heterolepis had the minimum DNA amount. A260/A280 purity degrees showed that the highest and lowest purity in gDNAs obtained through phenol-choloform isoamyl alcohol method were in I.aucheriana and I. salicina, respectively. The highest and lowest purity degrees of gDNAs obtained through commercial kit was observed in I. fragilis and I. macrocephala samples, respectively. PCR amplification results showed that while band profiles of each three regions (ITS, trnL-F and ndhF did not yield positive results in PCR amplifications using phenol-choloform isoamyl alcohol method; PCR band profiles obtained through commercial kit yielded positive results. As a result, it is fair to say that the relation of genomic DNA with PCR was found to be more efficient although the maximum amount of genomic DNA was obtained through phenol chloroform isoamyl alcohol method.

Evaluation of Etest and macrodilution broth method for antifungal susceptibility testing of Candida sp strains isolated from oral cavities of AIDS patients

Directory of Open Access Journals (Sweden)

SILVA Maria do Rosário R.

2002-01-01

Full Text Available A comparison of the Etest and the reference broth macrodilution susceptibility test for fluconazole, ketoconazole, itraconazole and amphotericin B was performed with 59 of Candida species isolated from the oral cavities of AIDS patients. The Etest method was performed according to the manufacturer's instructions, and the reference method was performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines. Our data showed that there was a good correlation between the MICs obtained by the Etest and broth dilution methods. When only the MIC results at ± 2 dilutions for both methods were considered, the agreement rates were 90.4% for itraconazole, ketoconazole and amphotericin B and 84.6% for fluconazole of the C. albicans tested. In contrast, to the reference method, the Etest method classified as susceptible three fluconazole-resistant isolates and one itraconazole-resistant isolate, representing four very major errors. These results indicate that Etest could be considered useful for antifungal sensitivity evaluation of yeasts in clinical laboratories.

In-situ reduced graphene oxide-polyvinyl alcohol composite coatings as protective layers on magnesium substrates

Directory of Open Access Journals (Sweden)

Xingkai Zhang

2017-06-01

Full Text Available A simple and feasible method was developed to fabricate in-situ reduced graphene oxide-polyvinyl alcohol composite (GO-PVA coatings as protective layers on magnesium substrates. Polyvinyl alcohol was used as an in-situ reductant to transform GO into reduced GO. Contiguous and uniform GO-PVA coatings were prepared on magnesium substrates by dip-coating method, and were further thermally treated at 120 °C under ambient condition to obtain in-situ reduced GO-PVA coatings. Owing to the reducing effect of PVA, thermal treatment at low temperature led to effective in-situ reduction of GO as confirmed by XRD, Raman, FTIR and XPS tests. The corrosion current density of magnesium substrates in 3.5 wt% NaCl solution could be lowered to its 1/25 when using in-situ reduced GO-PVA coatings as protective layers.

Responses of nutrients and mobile carbohydrates in Quercus variabilis seedlings to environmental variations using in situ and ex situ experiments.

Directory of Open Access Journals (Sweden)

Jing-Pin Lei

Full Text Available Forest tree species distributed across a wide range of geographical areas are subjected to differential climatic and edaphic conditions and long-term selection, leading to genotypes with morphological and physiological adaptation to the local environment. To test the ability of species to cope with changing environmental conditions, we studied the ecophysiological features of Quercus variabilis using seedlings grown in geographically widely isolated populations (Exp. I, in situ and in a common garden (Exp. II, ex situ using seedlings originating from those populations. We found that Q. variabilis plants grown in different locations along a south-north gradient had different levels of nutrients (N, P, K and carbon-physiological performance (photosynthesis, non-structural carbohydrates, such as soluble sugars and starch, and that these physiological differences were not correlated with local soil properties. These geographic variations of plant physiology disappeared when plants from different locations were grown in the same environment. Our results indicate that the physiological performance of Q. variabilis plants is mainly determined by the climatic variations across latitude rather than by their soils or by genetic differentiation. The adaptive ability of Q. variabilis found in the present study suggests that this species has the potential to cope, at least to some extent, with changing environmental conditions.

Survey of active and inactive mines for possible use as in situ test facilities

International Nuclear Information System (INIS)

1976-09-01

A survey of active and inactive mines which might be useful for radioactive waste storage in situ test experiments was conducted. It was performed for Union Carbide Corporation, Nuclear Division, Office of Waste Isolation. The report covers available information gathered from literature, U.S. Bureau of Mines, the Mining Enforcement and Safety Agency, and a limited number of personal contacts with constructors or operators of facilities. This survey is preliminary in nature and the objective is to develop potential candidate facilities for in situ experiments which warrant further investigation. Included are descriptions of 244 facilities, with all the data about each one which was available within the time restraint of the study. These facility descriptions are additionally indexed by depth of mine, nature of the country rock, mineral mined, and type of entry. A total of 14 inactive mines and 34 active mines has been selected as those most worthy of further investigation for possible service as nuclear waste isolation test facilities. This investigation, being preliminary and having been performed in a very short time period, must be qualified, and the description of the qualification is presented in the body of this report. Qualifications deal primarily with the hazard of having omitted facilities and having incomplete data in some instances. Results indicate sedimentary rock mines of minerals of evaporite origin as a first ranking of preference for in situ testing, followed by other sedimentary rocks and then by mines producing minerals from any type rock where the mine is above the local water table. These are general rules and of course there can be exceptions to them

In situ deposition of thallium-containing oxides

International Nuclear Information System (INIS)

Myers, K.E.

1996-01-01

The number and variety of thallium based materials that can be made by in situ methods have grown consistently since the first report of successful thallium cuprate deposition by Face and Nestlerode in 1992. Processes for the deposition of superconductors, normal metals, and insulators have been developed. Most work to date has been done on the Tl-1212 phases, TlBa 2 CaCu 2 O 7 and (Tl,Pb)Sr 2 CaCu 2 O 7 . Recently however, the in situ thallium technique has been extended to other materials. For example, epitaxial thin films of thallium tantalate, an insulator of the pyrochlore structure and a potential buffer layer for thallium cuprate films, have been grown. Multilayers, important in the fabrication of Josephson junctions, have been demonstrated with the thallium lead cuprates. This paper reviews progress in the area of in situ thallium deposition technology which will make more complex thallium cuprate multilayer structures and devices possible

Comparative study of different methods used to isolate Trypanosoma cruz i for defibrinated blood of irradiated mice

International Nuclear Information System (INIS)

Fontes, G.; Romanha, A.J.; Brener, Z.

1991-01-01

Different methods are being used for the isolation and purification of Trypanosoma cruzi blood forms from infected vertebrate hosts. In this study, we compare four of these methods (differential centrifugation, Ficoll-Hypaque, Histopaque 1077 and metrizamide) in terms of parasite recovery rates, contamination with cells, duration of the process and role of host irradiation. Male albino Swiss mice irradiated in a Gamma Cell 220(500 rads) were inoculated with C L and V-10 T.cruzi strains and bled at the peak of parasitemia. Infected defibrinated blood was then used for the isolation. Although all methods permitted the recovery of viable trypomastigotes, the best results were obtained with Ficoll-Hypaque and Histopaque 1077. Recovery rates ranged between 71% to 88% and parasite-enriched preparations were obtained in approximately 75 min. Irradiation and blood defibrination drastically reduced platelet and leukocyte contamination of the preparations. (author)

Deformation processes in functional materials studied by in situ neutron diffraction and ultrasonic techniques

International Nuclear Information System (INIS)

Sittner, P.; Novak, V.; Landa, M.; Lukas, P.

2007-01-01

The unique thermomechanical functions of shape memory alloys (hysteretic stress-strain-temperature responses) not their structural properties (as strength, fatigue, corrosion resistance, etc.) are primarily utilized in engineering applications. In order to better understand and predict the functional behavior, we have recently employed two dedicated non-invasive in situ experimental methods capable to follow the deformation/transformation processes in thermomechanically loaded polycrystalline samples. The in situ neutron diffraction method takes advantage of the ability of thermal neutrons to penetrate bulk samples. As a diffraction technique sensitive to interplanar spacings in crystalline solids, it provides in situ information on the changes in crystal structure, phase composition, phase stress and texture in the transforming samples. The combined in situ ultrasonic and electric resistance method follows variations of the electric resistance as well as speed and attenuation of acoustic waves propagating through the transforming sample. The acoustic waves are mainly sensitive to changes of elastic properties accompanying the deformation/transformation processes. The latter method thus follows the changes in interatomic bonds rather than changes in the interplanar lattice spacings focused in the neutron diffraction method. The methods are thus complementary. They are briefly described and selected experimental results obtained recently on NiTi alloys are presented and discussed

Development of an integrated, in-situ remediation technology. Topical report for task No. 9. Part I. TCE degradation using nonbiological methods, September 26, 1994--May 25, 1996

International Nuclear Information System (INIS)

Shapiro, A.P.; Sivavec, T.M.; Baghel, S.S.

1997-01-01

Contamination in low-permeability soils poses a significant technical challenge for in situ remediation efforts. Poor accessibility to the contaminants and difficulty in delivery of treatment reagents have rendered existing in situ treatments such as bioremediation, vapor extraction, pump and treat rather ineffective when applied to low-permeability soils present at many contaminated sites. The technology is an integrated in situ treatment in which established geotechnical methods are used to install degradation zones directly in the contaminated soil and electro-osmosis is used to move the contaminants back and forth through those zones until the treatment is completed. The present Draft Topical Report for Task No. 9 summarizes laboratory investigations into TCE degradation using nonbiological methods. These studies were conducted by the General Electric Company. The report concentrates on zero valent iron as the reducing agent and presents data on TCE and daughter product degradation rates in batch experiments, column studies, and electroosmotic cells. It is shown that zero valent iron effectively degrades TCE in electroosmotic experiments. Daughter product degradation and gas generation are shown to be important factors in designing field scale treatment zones for the Lasagna trademark process

In situ calibration of acoustic emission transducers by time reversal method

Czech Academy of Sciences Publication Activity Database

Kober, Jan; Převorovský, Zdeněk; Chlada, Milan

2016-01-01

Roč. 240, April (2016), s. 50-56 ISSN 0924-4247 Institutional support: RVO:61388998 Keywords : time reversed acoustic s * calibration * in situ * acoustic emission Subject RIV: BI - Acoustic s Impact factor: 2.499, year: 2016 http://ac.els-cdn.com/S0924424716300334/1-s2.0-S0924424716300334-main.pdf?_tid=0acf4736-ef6d-11e5-b826-00000aacb362&acdnat=1458568911_1c21eda9762b905a684ff939463ef3fe

A screening method for the isolation of polyhydroxyalkanoate-producing purple non-sulfur photosynthetic bacteria from natural seawater

Directory of Open Access Journals (Sweden)

Mieko Higuchi-Takeuchi

2016-09-01

Full Text Available Polyhydroxyalkanoates (PHAs are a family of biopolyesters accumulated by a variety of microorganisms as carbon and energy storage under starvation conditions. We focused on marine purple non-sulfur photosynthetic bacteria as host microorganisms for PHA production and developed a method for their isolation from natural seawater. To identify novel PHA-producing marine purple non-sulfur photosynthetic bacteria, natural seawaters were cultured in nutrient-rich medium for purple non-sulfur photosynthetic bacteria, and twelve pink- or red-pigmented colonies were picked up. Gas chromatography mass spectrometry analysis revealed that four isolates synthesized PHA at levels ranging from 0.5 to 24.4 wt% of cell dry weight. The 16S ribosomal RNA sequence analysis revealed that one isolate (HM2 showed 100% identity to marine purple non-sulfur photosynthetic bacteria. In conclusion, we have demonstrated in this study that PHA-producing marine purple non-sulfur photosynthetic bacteria can be isolated from natural seawater under nutrient-rich conditions.

Total error components - isolation of laboratory variation from method performance

International Nuclear Information System (INIS)

Bottrell, D.; Bleyler, R.; Fisk, J.; Hiatt, M.

1992-01-01

The consideration of total error across sampling and analytical components of environmental measurements is relatively recent. The U.S. Environmental Protection Agency (EPA), through the Contract Laboratory Program (CLP), provides complete analyses and documented reports on approximately 70,000 samples per year. The quality assurance (QA) functions of the CLP procedures provide an ideal data base-CLP Automated Results Data Base (CARD)-to evaluate program performance relative to quality control (QC) criteria and to evaluate the analysis of blind samples. Repetitive analyses of blind samples within each participating laboratory provide a mechanism to separate laboratory and method performance. Isolation of error sources is necessary to identify effective options to establish performance expectations, and to improve procedures. In addition, optimized method performance is necessary to identify significant effects that result from the selection among alternative procedures in the data collection process (e.g., sampling device, storage container, mode of sample transit, etc.). This information is necessary to evaluate data quality; to understand overall quality; and to provide appropriate, cost-effective information required to support a specific decision

Recent advances in micro-vibration isolation

Science.gov (United States)

Liu, Chunchuan; Jing, Xingjian; Daley, Steve; Li, Fengming

2015-05-01

Micro-vibration caused by disturbance sources onboard spacecraft can severely degrade the working environment of sensitive payloads. Some notable vibration control methods have been developed particularly for the suppression or isolation of micro-vibration over recent decades. Usually, passive isolation techniques are deployed in aerospace engineering. Active isolators, however, are often proposed to deal with the low frequency vibration that is common in spacecraft. Active/passive hybrid isolation has also been effectively used in some spacecraft structures for a number of years. In semi-active isolation systems, the inherent structural performance can be adjusted to deal with variation in the aerospace environment. This latter approach is potentially one of the most practical isolation techniques for micro-vibration isolation tasks. Some emerging advanced vibration isolation methods that exploit the benefits of nonlinearity have also been reported in the literature. This represents an interesting and highly promising approach for solving some challenging problems in the area. This paper serves as a state-of-the-art review of the vibration isolation theory and/or methods which were developed, mainly over the last decade, specifically for or potentially could be used for, micro-vibration control.

Oxygen-free in situ scanning tunnelling microscopy

DEFF Research Database (Denmark)

Zhang, Jingdong; Ulstrup, Jens

2007-01-01

Scanning tunneling microscopy under full electrochemical potential control (in situ STM) has been used extensively as an efficient method to characterize microstructures at solid/liquid interfaces at the atomic and molecular levels. However, under ambient conditions oxygen may interfere in open i...

Identification of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in Blood Cultures: a Multicenter Performance Evaluation of a Three-Color Peptide Nucleic Acid Fluorescence In Situ Hybridization Assay▿

Science.gov (United States)

Della-Latta, Phyllis; Salimnia, Hossein; Painter, Theresa; Wu, Fann; Procop, Gary W.; Wilson, Deborah A.; Gillespie, Wendy; Mender, Alayna; Crystal, Benjamin S.

2011-01-01

A multicenter evaluation was undertaken to evaluate the performance of a new three-color peptide nucleic acid fluorescence in situ hybridization assay that identifies isolates directly from blood cultures positive for Gram-negative bacilli (GNB). The assay correctly identified 100% (186/186) of the Escherichia coli isolates, 99.1% (109/110) of the Klebsiella pneumoniae isolates, and 95.8% (46/48) of the Pseudomonas aeruginosa isolates in this study. Negative assay results were correctly obtained for 162 of 165 other GNB (specificity, 98.2%). PMID:21490185

OpenMC In Situ Source Convergence Detection

Energy Technology Data Exchange (ETDEWEB)

Aldrich, Garrett Allen [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Univ. of California, Davis, CA (United States); Dutta, Soumya [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); The Ohio State Univ., Columbus, OH (United States); Woodring, Jonathan Lee [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

2016-05-07

We designed and implemented an in situ version of particle source convergence for the OpenMC particle transport simulator. OpenMC is a Monte Carlo based-particle simulator for neutron criticality calculations. For the transport simulation to be accurate, source particles must converge on a spatial distribution. Typically, convergence is obtained by iterating the simulation by a user-settable, fixed number of steps, and it is assumed that convergence is achieved. We instead implement a method to detect convergence, using the stochastic oscillator for identifying convergence of source particles based on their accumulated Shannon Entropy. Using our in situ convergence detection, we are able to detect and begin tallying results for the full simulation once the proper source distribution has been confirmed. Our method ensures that the simulation is not started too early, by a user setting too optimistic parameters, or too late, by setting too conservative a parameter.

Automated methods for single-stranded DNA isolation and dideoxynucleotide DNA sequencing reactions on a robotic workstation

International Nuclear Information System (INIS)

Mardis, E.R.; Roe, B.A.

1989-01-01

Automated procedures have been developed for both the simultaneous isolation of 96 single-stranded M13 chimeric template DNAs in less than two hours, and for simultaneously pipetting 24 dideoxynucleotide sequencing reactions on a commercially available laboratory workstation. The DNA sequencing results obtained by either radiolabeled or fluorescent methods are consistent with the premise that automation of these portions of DNA sequencing projects will improve the reproducibility of the DNA isolation and the procedures for these normally labor-intensive steps provides an approach for rapid acquisition of large amounts of high quality, reproducible DNA sequence data

Antimicrobial susceptibility determined by the E test, Löwenstein-Jensen proportion, and DNA sequencing methods among Mycobacterium tuberculosis isolates discrepancies, preliminary results

Directory of Open Access Journals (Sweden)

Maria Inês Moura Freixo

2004-02-01

Full Text Available Mycobacterium tuberculosis strains resistant to streptomycin (SM, isoniazid (INH, and/or rifampin (RIF as determined by the conventional Löwenstein-Jensen proportion method (LJPM were compared with the E test, a minimum inhibitory concentration susceptibility method. Discrepant isolates were further evaluated by BACTEC and by DNA sequence analyses for mutations in genes most often associated with resistance to these drugs (rpsL, katG, inhA, and rpoB. Preliminary discordant E test results were seen in 75% of isolates resistant to SM and in 11% to INH. Discordance improved for these two drugs (63% for SM and none for INH when isolates were re-tested but worsened for RIF (30%. Despite good agreement between phenotypic results and sequencing analyses, wild type profiles were detected on resistant strains mainly for SM and INH. It should be aware that susceptible isolates according to molecular methods might contain other mechanisms of resistance. Although reproducibility of the LJPM susceptibility method has been established, variable E test results for some M. tuberculosis isolates poses questions regarding its reproducibility particularly the impact of E test performance which may vary among laboratories despite adherence to recommended protocols. Further studies must be done to enlarge the evaluated samples and looked possible mutations outside of the hot spot sequenced gene among discrepant strains.

Species identification of Candida isolated from clinical specimens in a tertiary care hospital

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lsmet Nigar

2016-07-01

Full Text Available Background: Candida species are responsible for various clinical manifestations from mucocutaneous overgrowth to blood stream infections especially in immunocompromized situations. Although C. albicans is the most prevalent species, high incidence of non-albicans Candida species with antifungal resistance are emerging which is posing a serious threat to the patients care.Objective: This study aimed to isolate and identify different species of Candida from different clinical specimens. Methods: A total of 100 different clinical specimens were studied of which 35 were oral swab, 28 were high vaginal swab, 15 were urine, 14 were nail, 04 were bronchoalveolar lavage and peritoneal fluid were 04. Among 100 clinical specimens, Candida isolates were identified in 64 specimens. Isolation of Candida species was done by primary culture in SDA. Subsequent identification of species were performed by germ tube test, subculture in chromo­genic agar medium and carbohydrate assimilation test with commonly used twelve sugars.Results: Out of 64 isolated Candida species, Candida albicans were 51.56% and the non-albicans Candida species were 48.44%. The most prevalent Candida species was C. albicans 33 (51.53% followed by C. tropicalis 17 (26.56%. C. glabrata 4 (6.25%, C. parapsilo­sis 4 (6.25%, C. krusei 3 (4.68% and C. guilliermondii 2 (3.2%. One of the isolated Candida species was unidentified.Conclusion: Though Candida albicans was found as the most common species, but non-albicans Candida species are appearing as emerging pathogens as well. Exposure to chemotherapy appeared to be the commonest predisposing factor for Candida infection followed by indwelling urinary catheter in situ for prolong period.

In situ shape and distance measurements in neutron scattering and diffraction

Energy Technology Data Exchange (ETDEWEB)

Fujiwara, Satoru; Mendelson, R.A. [Univ. of California, San Francisco, CA (United States)

1994-12-31

Neutron scattering combined with selective isotopic labeling and contrast matching is useful for obtaining in situ structural information about a selected particle, or particles, in a macromolecular complex. The observed intensities, however, may be distorted by inter-complex interference and by scattering-length-density fluctuations of the (otherwise) contrast-matched portions. Methods have been proposed to cancel out such distortions (Hoppe`s method, the Statistical Labeling Method, and the Triple Isotopic Substitution Method). With these methods as well as related unmixed-sample methods, structural information about the selected particles can be obtained without these distortions. We have generalized these methods so that, in addition to globular particles in solution, they can be applied to in situ structures of systems having underlying symmetry and/or net orientation as well. The information obtainable from such experiments is discussed.

A novel method for isolation of epithelial cells from ovine esophagus for tissue engineering.

Science.gov (United States)

Macheiner, Tanja; Kuess, Anna; Dye, Julian; Saxena, Amulya K

2014-01-01

The yield of a critical number of basal epithelial cells with high mitotic rates from native tissue is a challenge in the field of tissue engineering. There are many protocols that use enzymatic methods for isolation of epithelial cells with unsatisfactory results for tissue engineering. This study aimed to develop a protocol for isolating a sufficient number of epithelial cells with a high Proliferating Index from ovine esophagus for tissue engineering applications. Esophageal mucosa was pretreated with dispase-collagenase solution and plated on collagen-coated culture dishes. Distinction of the various types of epithelial cells and developmental stages was done with specific primary antibodies to Cytokeratins and to Proliferating Cell Nuclear Antigen (PCNA). Up to approximately 8100 epithelial cells/mm2 of mucosa tissue were found after one week of migration. Cytokeratin 14 (CK 14) was positive identified in cells even after 83 days. At the same time the Proliferating Index was 71%. Our protocol for isolation of basal epithelial cells was successful to yield sufficient numbers of cells predominantly with proliferative character and without noteworthy negative enzymatic affection. The results at this study offer the possibility of generation critical cell numbers for tissue engineering applications.

Automatic speech recognition (zero crossing method). Automatic recognition of isolated vowels

International Nuclear Information System (INIS)

Dupeyrat, Benoit

1975-01-01

This note describes a recognition method of isolated vowels, using a preprocessing of the vocal signal. The processing extracts the extrema of the vocal signal and the interval time separating them (Zero crossing distances of the first derivative of the signal). The recognition of vowels uses normalized histograms of the values of these intervals. The program determines a distance between the histogram of the sound to be recognized and histograms models built during a learning phase. The results processed on real time by a minicomputer, are relatively independent of the speaker, the fundamental frequency being not allowed to vary too much (i.e. speakers of the same sex). (author) [fr

The interval high rate discharge behavior of Li3V2(PO4)3/C cathode based on in situ polymerization method

International Nuclear Information System (INIS)

Mao, Wen-feng; Yan, Ji; Xie, Hui; Tang, Zhi-yuan; Xu, Qiang

2013-01-01

An in situ polymerization assisted fast sol–gel method was introduced to synthesize high performance Li 3 V 2 PO 4 /C (LVP/C) cathode material. The crystal structure, surface morphology and electrochemical performances of the LVP/C samples sintered at different temperatures were investigated. The composite sintered at 750 °C exhibits the highest specific discharge capacity of 119.02 mAh g −1 (440.35 Wh g −1 ) at 10 C rate. The Li + diffusion coefficient ranges from 10 −6 to 10 −8 cm 2 s −1 based on different scanning rates and the electronic conductivity is about 10 −5 S cm −1 . For comparison, an ex situ polymerization method was also employed to obtain the LVP/C composite. A novel charge/discharge testing mode was designed to investigate the electrochemical behavior of the as-prepared LVP/C composite for practical application in electric vehicle cells. The obtained high power density and the special testing mode prove the LVP/C composite would be a promising candidate for the electric vehicle application and deserves further investigation

Colorimetric method for the detection of melamine using in-situ formed silver nanoparticles via tannic acid

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Alam, Md. Fazle; Laskar, Amaj Ahmed; Ahmed, Shahbaz; Shaida, Mohd. Azfar; Younus, Hina

2017-08-01

Melamine toxicity has recently attracted worldwide attention as it causes renal failure and the death of humans and animals. Therefore, developing a simple, fast and sensitive method for the routine detection of melamine is the need of the hour. Herein, we have developed a selective colorimetric method for the detection of melamine in milk samples based upon in-situ formation of silver nanoparticles (AgNPs) via tannic acid. The AgNPs thus formed were characterized by UV-Visible spectrophotometer, transmission electron microscope (TEM), zetasizer and dynamic light scattering (DLS). The AgNPs were used to detect melamine under in vitro condition and in raw milk spiked with melamine. Under optimal conditions, melamine could be selectively detected in vitro within the concentration range of 0.05-1.4 μM with a limit of detection (LOD) of 0.01 μM, which is lower than the strictest melamine safety requirement of 1 ppm. In spiked raw milk, the recovery percentage range was 99.5-106.5% for liquid milk and 98.5-105.5% for powdered milk. The present method shows extreme selectivity with no significant interference with other substances like urea, glucose, glycine, ascorbic acid etc. This assay method does not utilize organic cosolvents, enzymatic reactions, light sensitive dye molecules and sophisticated instrumentation, thereby overcoming some of the limitations of the other conventional methods.

A microfluidic membrane chip for in situ fouling characterization

NARCIS (Netherlands)

Ngene, I.S.; Lammertink, Rob G.H.; Wessling, Matthias; van der Meer, Walterus Gijsbertus Joseph

2010-01-01

A new method for non-invasive in situ monitoring of a microfiltration process is described. In microfiltration systems, local information on the deposition characteristics can be used to determine the cake behavior during a filtration run. Typically, non-invasive methods of fouling study are

Comparison of vapor sampling system (VSS) and in situ vapor sampling (ISVS) methods on Tanks C-107, BY-108, and S-102. Revision 1

International Nuclear Information System (INIS)

Huckaby, J.L.; Edwards, J.A.; Evans, J.C.

1996-08-01

This report discusses comparison tests for two methods of collecting vapor samples from the Hanford Site high-level radioactive waste tank headspaces. The two sampling methods compared are the truck-mounted vapor sampling system (VSS) and the cart-mounted in-situ vapor sampling (ISVS). Three tanks were sampled by both the VSS and ISVS methods from the same access risers within the same 8-hour period. These tanks have diverse headspace compositions and they represent the highest known level of several key vapor analytes

A novel tertiary prep-HPLC method for the isolation of single amino acids for AMS-radiocarbon measurement.

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Fernandes, Ricardo; Koudelka, Tomas; Tholey, Andreas; Dreves, Alexander

2017-07-15

AMS-radiocarbon measurements of amino acids can potentially provide more reliable radiocarbon dates than bulk collagen analysis. Nonetheless, the applicability of such an approach is often limited by the low-throughput of existing isolation methods and difficulties in determining the contamination introduced during the separation process. A novel tertiary prep-HPLC amino acid isolation method was developed that relies on the combustion of eluted material without requiring any additional chemical steps. Amino acid separation was carried out using a gradient mix of pure water and phosphoric acid with an acetonitrile step in-between runs to remove hydrophobic molecules from the separation column. The amount of contaminant carbon and its 14 C content were determined from two-point measurements of collagen samples of known 14 C content. The amount of foreign carbon due to the isolation process was estimated at 4±1μg and its 14 C content was 0.43±0.01 F 14 C. Radiocarbon values corrected for carbon contamination have only a minor increase in uncertainties. For Holocene samples, this corresponds to an added uncertainty typically smaller than 10 14 Cyears. The developed method can be added to routine AMS measurements without implying significant operational changes and offers a level of measurement uncertainty that is suitable for many archaeological, ecological, environmental, and biological applications. Copyright © 2017. Published by Elsevier B.V.

Identification of Mucorales isolates from soil using morphological and molecular methods.

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Ziaee, A; Zia, M; Bayat, M; Hashemi, J

2016-03-01

Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera Rhizopus , Mucor , Rhizomucor , Lichtheimia ( Absidia) , Cunninghamella, and Mortierella . Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features . In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies. 400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera Lichtheimia , Rhizopus, Rhizomucor, Mucor, Cunninghamella, and Mortierella, were identified . The genus Rhizopus (35.5%) was the most frequent isolate, followed by Mucor (32.25%) and Rhizomucor (27.5%). The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals.

Isolation, characterization, antibiogram and pathology of Pasteurella multocida isolated from pigs

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Mamta Tigga

2014-05-01

Full Text Available Aim: Isolation, characterization and antibiogram of Pasteurella multocida from diseased pigs of district Durg of Chhattisgarh, and to study pathological changes caused by swine pasteurellosis. Materials and Methods: An outbreak of swine pasteurellosis was suspected in pigs of Ruwabandha (Bhilai, Anjora, Somni, Tedesara, Tirgajhola villages of Durg district in Chhattisgarh, India during August and September of 2011. Nasal Swabs and blood samples from ailing pigs and heart blood and impression smears from morbid pigs were processed for detection and isolation of P. multocida by bacteriological methods. Detailed necropsy was conducted and gross and histopathological lesions were recorded. The test Isolates were subjected to antimicrobial sensitivity profile by disc-diffusion method. Results: The blood smears from heart blood and tissue impression smears revealed teaming of bipolar organisms indicating the presence of Pasteurella spp. The isolates obtained were subjected to Gram's staining for checking the purity and bipolar morphology and characterized biochemically. Gross lesions included severe acute pneumonia and haemorrhages in lungs, petechial haemorrhages on serous membranes and other visceral organs. On histopathological examination, lungs showed typical fibrinous bronchopneumonia, multifocal suppuration. All the isolates of P. multocida were 100% sensitive to Amoxicillin, Gentamicin, Enrofloxacin and showed100% resistance to Ceftizoxim and Cloxacillin. Conclusion: Gross and microscopic lesions in dead animals are of great diagnostic value and are of characteristic of P. multocida infection. Cultural, morphological and biochemical characters are useful to rule out the causative agent as P. multocida. Antibiotic sensitivity pattern of the isolates should routinely be carried out for knowing the antibiotic resistance trends in an endemic area.

Supercapacitor electrode of nano-Co3O4 decorated with gold nanoparticles via in-situ reduction method

Science.gov (United States)

Tan, Yongtao; Liu, Ying; Kong, Lingbin; Kang, Long; Ran, Fen

2017-09-01

Nano-Co3O4 decorated with gold nanoparticles is synthesized by a simple method of in-situ reduction of HAuCl4 by sodium citrate for energy storage application, and the effect of gold content in the product on electrochemical performance is investigated in detail. Introducing gold nanoparticles into nano-Co3O4 bulk would contribute to reduce internal resistance of charge transmission. The results show that after in-situ reduction reaction gold nanoparticles imbed uniformly into nano-Co3O4 with irregular nanoparticles. The gold nanoparticles decorated nano-Co3O4 exhibits specific capacitance of 681 F g-1 higher than that of pristine Co3O4 of 368 F g-1. It is interesting that a good cycle life with the specific capacitance retention of 83.1% is obtained after 13000 cycles at 5 A g-1, which recovers to initial specific capacitance value when the test current density is turned to 2 A g-1. In addition, the device of asymmetric supercapacitor, assembled with gold nanoparticles decorated nano-Co3O4 as the positive electrode and activated carbon as the negative electrode, exhibits good energy density of 25 Wh kg-1, which is comparable to the asymmetric device assembled with normal nano-Co3O4, or the symmetric device assembled just with activated carbon.

Automatic method for estimation of in situ effective contact angle from X-ray micro tomography images of two-phase flow in porous media.

Science.gov (United States)

Scanziani, Alessio; Singh, Kamaljit; Blunt, Martin J; Guadagnini, Alberto

2017-06-15

Multiphase flow in porous media is strongly influenced by the wettability of the system, which affects the arrangement of the interfaces of different phases residing in the pores. We present a method for estimating the effective contact angle, which quantifies the wettability and controls the local capillary pressure within the complex pore space of natural rock samples, based on the physical constraint of constant curvature of the interface between two fluids. This algorithm is able to extract a large number of measurements from a single rock core, resulting in a characteristic distribution of effective in situ contact angle for the system, that is modelled as a truncated Gaussian probability density distribution. The method is first validated on synthetic images, where the exact angle is known analytically; then the results obtained from measurements within the pore space of rock samples imaged at a resolution of a few microns are compared to direct manual assessment. Finally the method is applied to X-ray micro computed tomography (micro-CT) scans of two Ketton cores after waterflooding, that display water-wet and mixed-wet behaviour. The resulting distribution of in situ contact angles is characterized in terms of a mixture of truncated Gaussian densities. Crown Copyright © 2017. Published by Elsevier Inc. All rights reserved.

Isolation of uv-sensitive variants of human FL cells by a viral suicide method

International Nuclear Information System (INIS)

Shiomi, T.; Sato, K.

1979-01-01

A new method (viral suicide method) for the isolation of uv-sensitive mutants is described. Colonies of mutagenized human FL cells were infected with uv-irradiated Herpes simplex viruses and surviving ones which seemed to be deficient in host cell reactivation (HCR) were examined for their uv sensitivity. Nineteen of 238 clones examined were sensitive to uv irradiation at the time of the isolation. After recloning, four of these clones have been studied and two (UVS-1 and UVS-2) of them are stable in their uv sensitivity for 4 months in culture. uv sensitivity of UVS-1, UVS-2, and the parental FL cells are as follows: the extrapolation numbers (n) are 2.2, 2.1, and 1.8 and mean lethal doses (DO) are 2.9, 3.7, and 7.8 J/m 2 for UVS-1, UVS-2, and the parental FL cells, respectively. They are no more sensitive than FL cells to x-irradiation. The ability of HCR in UVS-2 cells is apparently lower than that in FL cells, whereas UVS-1 cells are the same as FL cells in the ability

Cell fusion in tumor progression: the isolation of cell fusion products by physical methods

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Vincitorio Massimo

2011-09-01

Full Text Available Abstract Background Cell fusion induced by polyethylene glycol (PEG is an efficient but poorly controlled procedure for obtaining somatic cell hybrids used in gene mapping, monoclonal antibody production, and tumour immunotherapy. Genetic selection techniques and fluorescent cell sorting are usually employed to isolate cell fusion products, but both procedures have several drawbacks. Results Here we describe a simple improvement in PEG-mediated cell fusion that was obtained by modifying the standard single-step procedure. We found that the use of two PEG undertreatments obtains a better yield of cell fusion products than the standard method, and most of these products are bi- or trinucleated polykaryocytes. Fusion rate was quantified using fluorescent cell staining microscopy. We used this improved cell fusion and cell isolation method to compare giant cells obtained in vitro and giant cells obtained in vivo from patients with Hodgkin's disease and erythroleukemia. Conclusions In the present study we show how to improve PEG-mediated cell fusion and that cell separation by velocity sedimentation offers a simple alternative for the efficient purification of cell fusion products and to investigate giant cell formation in tumor development.

Impact of isolation method on doubling time and the quality of chondrocyte and osteoblast differentiated from murine dental pulp stem cells

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Rohaya Megat Abdul Wahab

2017-06-01

Full Text Available Background Stem cells are normally isolated from dental pulps using the enzymatic digestion or the outgrowth method. However, the effects of the isolation method on the quality of the isolated stem cells are not studied in detail in murine models. The aim of this study was to compare the matrices secreted by osteoblast and chondrocytes differentiated from dental pulp stem cells isolated through different means. Method DPSC from murine incisors were isolated through either the outgrowth (DPSC-OG or the enzymatic digestion (DPSC-ED method. Cells at passage 4 were used in this study. The cells were characterized through morphology and expression of cell surface markers. The cells’ doubling time when cultured using different seeding densities was calculated and analyzed using one-way ANOVA and Tukey’s multiple comparison post-test. The ability of cells to differentiate to chondrocyte and osteoblast was evaluated through staining and analysis on the matrices secreted. Results Gene expression analysis showed that DPSC-OG and DPSC-ED expressed dental pulp mesenchymal stem cell markers, but not hematopoietic stem cell markers. The least number of cells that could have been used to culture DPSC-OG and DPSC-ED with the shortest doubling time was 5 × 102 cells/cm2 (11.49 ± 2.16 h and 1 × 102 cells/cm2 (10.55 h ± 0.50, respectively. Chondrocytes differentiated from DPSC-ED produced  2 times more proteoglycan and at a faster rate than DPSC-OG. FTIR revealed that DPSC-ED differentiated into osteoblast also secreted matrix, which more resembled a calvaria. Discussion Isolation approaches might have influenced the cell populations obtained. This, in turn, resulted in cells with different proliferation and differentiation capability. While both DPSC-OG and DPSC-ED expressed mesenchymal stem cell markers, the percentage of cells carrying each marker might have differed between the two methods. Regardless, enzymatic digestion clearly yielded cells

Operational Review of the First Wireline In Situ Stress Test in Scientific Ocean Drilling

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Casey Moore

2012-04-01

Full Text Available Scientific ocean drilling’s first in situ stress measurement was made at Site C0009A during Integrated Ocean Drilling Program (IODP Expedition 319 as part of Nankai Trough Seismogenic Zone Experiment (NanTroSEIZE Stage 2. The Modular Formation Dynamics Tester (MDT, Schlumbergerwireline logging tool was deployed in riser Hole C0009A to measure in situ formation pore pressure, formation permeability (often reported as mobility=permeability/viscosity, and the least principal stress (S3 at several isolated depths (Saffer et al., 2009; Expedition 319 Scientists, 2010. The importance of in situ stress measurements is not only for scientific interests in active tectonic drilling, but also for geomechanical and well bore stability analyses. Certain in situ tools were not previously available for scientific ocean drilling due to the borehole diameter and open hole limits of riserless drilling. The riser-capable drillship, D/V Chikyu,now in service for IODP expeditions, allows all of the techniques available to estimate the magnitudes and orientations of 3-D stresses to be used. These techniques include downhole density logging for vertical stress, breakout and caliper log analyses for maximum horizontal stress, core-based anelastic strain recovery (ASR, used in the NanTroSEIZE expeditions in 2007–2008, and leak-off test (Lin et al., 2008 and minifrac/hydraulic fracturing (NanTroSEIZE Expedition319 in 2009. In this report, the whole operational planning process related to in situ measurements is reviewed, and lessons learned from Expedition 319 are summarized for efficient planning and testing in the future.

In Situ Remediation Integrated Program: Technology summary

International Nuclear Information System (INIS)

1994-02-01

The In Situ Remediation Integrated Program (ISR IP) was instituted out of recognition that in situ remediation could fulfill three important criteria: significant cost reduction of cleanup by eliminating or minimizing excavation, transportation, and disposal of wastes; reduced health impacts on workers and the public by minimizing exposure to wastes during excavation and processing; and remediation of inaccessible sites, including: deep subsurfaces, in, under, and around buildings. Buried waste, contaminated soils and groundwater, and containerized wastes are all candidates for in situ remediation. Contaminants include radioactive wastes, volatile and non-volatile organics, heavy metals, nitrates, and explosive materials. The ISR IP intends to facilitate development of in situ remediation technologies for hazardous, radioactive, and mixed wastes in soils, groundwater, and storage tanks. Near-term focus is on containment of the wastes, with treatment receiving greater effort in future years. ISR IP is an applied research and development program broadly addressing known DOE environmental restoration needs. Analysis of a sample of 334 representative sites by the Office of Environmental Restoration has shown how many sites are amenable to in situ remediation: containment--243 sites; manipulation--244 sites; bioremediation--154 sites; and physical/chemical methods--236 sites. This needs assessment is focused on near-term restoration problems (FY93--FY99). Many other remediations will be required in the next century. The major focus of the ISR EP is on the long term development of permanent solutions to these problems. Current needs for interim actions to protect human health and the environment are also being addressed

In Situ Remediation Integrated Program: Technology summary

Energy Technology Data Exchange (ETDEWEB)

1994-02-01

The In Situ Remediation Integrated Program (ISR IP) was instituted out of recognition that in situ remediation could fulfill three important criteria: significant cost reduction of cleanup by eliminating or minimizing excavation, transportation, and disposal of wastes; reduced health impacts on workers and the public by minimizing exposure to wastes during excavation and processing; and remediation of inaccessible sites, including: deep subsurfaces, in, under, and around buildings. Buried waste, contaminated soils and groundwater, and containerized wastes are all candidates for in situ remediation. Contaminants include radioactive wastes, volatile and non-volatile organics, heavy metals, nitrates, and explosive materials. The ISR IP intends to facilitate development of in situ remediation technologies for hazardous, radioactive, and mixed wastes in soils, groundwater, and storage tanks. Near-term focus is on containment of the wastes, with treatment receiving greater effort in future years. ISR IP is an applied research and development program broadly addressing known DOE environmental restoration needs. Analysis of a sample of 334 representative sites by the Office of Environmental Restoration has shown how many sites are amenable to in situ remediation: containment--243 sites; manipulation--244 sites; bioremediation--154 sites; and physical/chemical methods--236 sites. This needs assessment is focused on near-term restoration problems (FY93--FY99). Many other remediations will be required in the next century. The major focus of the ISR EP is on the long term development of permanent solutions to these problems. Current needs for interim actions to protect human health and the environment are also being addressed.

A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning.

Science.gov (United States)

Carbonetti, Sara; Oliver, Brian G; Vigdorovich, Vladimir; Dambrauskas, Nicholas; Sack, Brandon; Bergl, Emilee; Kappe, Stefan H I; Sather, D Noah

2017-09-01

Monoclonal antibody technologies have enabled dramatic advances in immunology, the study of infectious disease, and modern medicine over the past 40years. However, many monoclonal antibody discovery procedures are labor- and time-intensive, low efficiency, and expensive. Here we describe an optimized mAb discovery platform for the rapid and efficient isolation, cloning and characterization of monoclonal antibodies in murine systems. In this platform, antigen-binding splenic B cells from immunized mice are isolated by FACS and cocultured with CD40L positive cells to induce proliferation and mAb production. After 12days of coculture, cell culture supernatants are screened for antigen, and IgG positivity and RNA is isolated for reverse-transcription. Positive-well cDNA is then amplified by PCR and the resulting amplicons can be cloned into ligation-independent expression vectors, which are then used directly to transfect HEK293 cells for recombinant antibody production. After 4days of growth, conditioned medium can be screened using biolayer interferometry for antigen binding and affinity measurements. Using this method, we were able to isolate six unique, functional monoclonal antibodies against an antigen of the human malaria parasite Plasmodium falciparum. Importantly, this method incorporates several important advances that circumvent the need for single-cell PCR, restriction cloning, and large scale protein production, and can be applied to a wide array of protein antigens. Copyright © 2017 Elsevier B.V. All rights reserved.

Using the in situ lift-out technique to prepare TEM specimens on a single-beam FIB instrument

International Nuclear Information System (INIS)

Lekstrom, M; McLachlan, M A; Husain, S; McComb, D W; Shollock, B A

2008-01-01

Transmission electron microscope (TEM) specimens are today routinely prepared using focussed ion beam (FIB) instruments. Specifically, the lift-out method has become an increasingly popular technique and involves removing thin cross-sections from site-specific locations and transferring them to a TEM grid. This lift-out process can either be performed ex situ or in situ. The latter is mainly carried out on combined dual-beam FIB and scanning electron microscope (SEM) systems whereas conventional single-beam instruments often are limited to the traditional ex situ method. It is nevertheless desirable to enhance the capabilities of existing single-beam instruments to allow for in situ lift-out preparation to be performed since this technique offers a number of advantages over the older ex situ method. A single-beam FIB instrument was therefore modified to incorporate an in situ micromanipulator fitted with a tungsten needle, which can be attached to a cut-out FIB section using ion beam induced platinum deposition. This article addresses the issues of using an ion beam to monitor the in situ manipulation process as well as approaches that can be used to create stronger platinum welds between two objects, and finally, views on how to limit the extent of ion beam damage to the specimen surface.

Location specific in situ TEM straining specimens made using FIB

International Nuclear Information System (INIS)