Protein blotting protocol for beginners.
Petrasovits, Lars A
2014-01-01
The transfer and immobilization of biological macromolecules onto solid nitrocellulose or nylon (polyvinylidene difluoride (PVDF)) membranes subsequently followed by specific detection is referred to as blotting. DNA blots are called Southerns after the inventor of the technique, Edwin Southern. By analogy, RNA blots are referred to as northerns and protein blots as westerns (Burnette, Anal Biochem 112:195-203, 1981). With few exceptions, western blotting involves five steps, namely, sample collection, preparation, separation, immobilization, and detection. In this chapter, protocols for the entire process from sample collection to detection are described.
Western Blotting of the Endocannabinoid System.
Wager-Miller, Jim; Mackie, Ken
2016-01-01
Measuring expression levels of G protein-coupled receptors (GPCRs) is an important step for understanding the distribution, function, and regulation of these receptors. A common approach for detecting proteins from complex biological systems is Western blotting. In this chapter, we describe a general approach to Western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose membranes, with a focus on detecting type 1 cannabinoid (CB1) receptors. When this technique is carefully used, specifically with validation of the primary antibodies, it can provide quantitative information on protein expression levels. Additional information can also be inferred from Western blotting such as potential posttranslational modifications that can be further evaluated by specific analytical techniques.
The Design of a Quantitative Western Blot Experiment
Directory of Open Access Journals (Sweden)
Sean C. Taylor
2014-01-01
Full Text Available Western blotting is a technique that has been in practice for more than three decades that began as a means of detecting a protein target in a complex sample. Although there have been significant advances in both the imaging and reagent technologies to improve sensitivity, dynamic range of detection, and the applicability of multiplexed target detection, the basic technique has remained essentially unchanged. In the past, western blotting was used simply to detect a specific target protein in a complex mixture, but now journal editors and reviewers are requesting the quantitative interpretation of western blot data in terms of fold changes in protein expression between samples. The calculations are based on the differential densitometry of the associated chemiluminescent and/or fluorescent signals from the blots and this now requires a fundamental shift in the experimental methodology, acquisition, and interpretation of the data. We have recently published an updated approach to produce quantitative densitometric data from western blots (Taylor et al., 2013 and here we summarize the complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting.
International Nuclear Information System (INIS)
Thompson, G.A.; Davies, H.M.; McDonald, N.
1985-01-01
A method termed product-selective blotting has been developed for screening large numbers of samples for enzyme activity. The technique is particularly well suited to detection of enzymes in native electrophoresis gels. The principle of the method was demonstrated by blotting samples from glutaminase or glutamate synthase reactions into an agarose gel embedded with ion-exchange resin under conditions favoring binding of product (glutamate) over substrates and other substances in the reaction mixture. After washes to remove these unbound substances, the product was measured using either fluorometric staining or radiometric techniques. Glutaminase activity in native electrophoresis gels was visualized by a related procedure in which substrates and products from reactions run in the electrophoresis gel were blotted directly into a resin-containing image gel. Considering the selective-binding materials available for use in the image gel, along with the possible detection systems, this method has potentially broad application
TLC blot (far-eastern blot) and its applications.
Taki, Takao; Gonzalez, Tania Valdes; Goto-Inoue, Naoko; Hayasaka, Takahiro; Setou, Mitsutoshi
2009-01-01
A simple method for transfer of lipids including phospholipids, glycolipids, and neutral lipids from a high-performance thin-layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, called TLC blot (far-eastern blot), is presented. Lipids separated on a HPTLC plate are blotted quantitatively. This procedure made it possible to purify individual lipids from a blotted membrane in a short time. Binding study, immunodetection, and mass spectrometric analysis are available for PVDF membrane. Furthermore, the world of molecular species imaging is opened by a scanning analysis with a combination of TLC blot and matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer (TLC-Blot/MALDI-TOF MS).
Villota, Guido E; Gomez, Diana I; Volcy, Michel; Franco, Andrés F; Cardona, Edgar A; Isaza, Rodrigo; Sanzón, Fernando; Teale, Judy M; Restrepo, Blanca I
2003-03-01
The detection of antibodies to Taenia solium metacestodes is very important in the differential diagnosis of neurocysticercosis (NCC). In this study, an electroimmunotransfer blot (EITB) assay that uses an elaborate protocol with metacestode glycoproteins as antigens was compared with two other Western blots that use glycoproteins obtained using simpler methods, including an eluate from a lectin column, or the vesicular fluid (VF) of the parasite. The concordance between the three assays was 91% in patients with active NCC and 100% in patients with suspected NCC and previous documentation of negative serology. The specificities for the Western blots and the EITB assay were 98% and 100%, respectively (98% concordance). These data suggest that the simplest of these immunoassays, the one that uses the VF of T. solium metacestodes in a Western blot format, can be reliably used for the serologic diagnosis of NCC in developing countries where access to the EITB assay is difficult.
Western blotting using chemiluminescent substrates.
Alegria-Schaffer, Alice
2014-01-01
Western blotting is a powerful and commonly used tool to identify and quantify a specific protein in a complex mixture (Towbin et al., 1979). The technique enables indirect detection of protein samples immobilized on a nitrocellulose or polyvinylidene fluoride (PVDF) membrane. Copyright © 2014 Elsevier Inc. All rights reserved.
Kurien, Biji T; Scofield, R Hal
2006-04-01
Western blotting (protein blotting or immunoblotting) is a powerful and important procedure for the immunodetection of proteins post-electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer. This review describes the various procedures that have been used to transfer proteins from a gel to a membrane based on the principles of simple diffusion, vacuum-assisted solvent flow and electrophoretic elution. Finally, a brief description of methods generally used to detect antigens on blots is also described.
Use of a Western blot technique for the serodiagnosis of glanders
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de Souza Marcilia MA
2011-01-01
Full Text Available Abstract Background The in vivo diagnosis of glanders relies on the highly sensitive complement fixation test (CFT. Frequently observed false positive results are troublesome for veterinary authorities and cause financial losses to animal owners. Consequently, there is an urgent need to develop a test with high specificity. Hence, a Western blot assay making use of a partly purified lipopolysaccaride (LPS containing antigen of three Burkholderia mallei strains was developed. The test was validated investigating a comprehensive set of positive and negative sera obtained from horses and mules from endemic and non endemic areas. Results The developed Western blot assay showed a markedly higher diagnostic specificity when compared to the prescribed CFT and therefore can be used as a confirmatory test. However, the CFT remains the test of choice for routine testing of glanders due to its high sensitivity, its feasibility using standard laboratory equipment and its worldwide distribution in diagnostic laboratories. Conclusions The CFT should be amended by the newly validated Western blot to increase the positive likelihood ratio of glanders serodiagnosis in non endemic areas or areas with low glanders prevalence. Its use for international trade of horses and mules should be implemented by the OIE.
Multistrip western blotting to increase quantitative data output.
Kiyatkin, Anatoly; Aksamitiene, Edita
2009-01-01
The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip western blotting increases the data output per single blotting cycle up to tenfold, allows concurrent monitoring of up to nine different proteins from the same loading of the sample, and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data, and therefore is beneficial to apply in biomedical diagnostics, systems biology, and cell signaling research.
Okabe, Masashi; Nyakas, Csaba; Buwalda, Bauke; Luiten, Paul G.M.
1993-01-01
We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility
Automated design of genomic Southern blot probes
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Komiyama Noboru H
2010-01-01
Full Text Available Abstract Background Sothern blotting is a DNA analysis technique that has found widespread application in molecular biology. It has been used for gene discovery and mapping and has diagnostic and forensic applications, including mutation detection in patient samples and DNA fingerprinting in criminal investigations. Southern blotting has been employed as the definitive method for detecting transgene integration, and successful homologous recombination in gene targeting experiments. The technique employs a labeled DNA probe to detect a specific DNA sequence in a complex DNA sample that has been separated by restriction-digest and gel electrophoresis. Critically for the technique to succeed the probe must be unique to the target locus so as not to cross-hybridize to other endogenous DNA within the sample. Investigators routinely employ a manual approach to probe design. A genome browser is used to extract DNA sequence from the locus of interest, which is searched against the target genome using a BLAST-like tool. Ideally a single perfect match is obtained to the target, with little cross-reactivity caused by homologous DNA sequence present in the genome and/or repetitive and low-complexity elements in the candidate probe. This is a labor intensive process often requiring several attempts to find a suitable probe for laboratory testing. Results We have written an informatic pipeline to automatically design genomic Sothern blot probes that specifically attempts to optimize the resultant probe, employing a brute-force strategy of generating many candidate probes of acceptable length in the user-specified design window, searching all against the target genome, then scoring and ranking the candidates by uniqueness and repetitive DNA element content. Using these in silico measures we can automatically design probes that we predict to perform as well, or better, than our previous manual designs, while considerably reducing design time. We went on to
TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics.
Taki, Takao
2015-01-01
A simple method for transfer of lipids-including phospholipids, glycolipids, and neutral lipids-from a high performance thin layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, TLC-Blot (Far-Eastern Blot), and its biochemical applications are presented. This chapter presents the conventional procedures for separating lipid from tissue samples, cultured cells, and serum and the subsequent development of TLC. Individual lipids separated on an HPTLC plate can be transferred to the PVDF membrane quantitatively and also isolated from the lipid-blotted membrane by a one-step purification procedure. Immunodetection with monoclonal antibodies and treatment with lipid-metabolizing enzymes on the lipid-blotted membrane are possible. The method for identification of individual lipids transferred on the PVDF membrane using matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (TLC-Blot/MALDI-TOF MS) is shown as a functional lipidomics application.
Southern, Edwin
2015-01-01
The history of the development of DNA blotting is described in this chapter. DNA blotting, involving the transfer of electrophoretically separated DNA fragments to a membrane support through capillary action, is also known as Southern blotting. This procedure enables the detection of a specific DNA sequence by hybridization with probes. The term Southern blotting led to a "geographic" naming tradition, with RNA blotting bearing the name Northern blotting and protein transfer to membranes becoming known as Western blotting.
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Josefsen, Knud; Nielsen, Henrik
2011-01-01
Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membrane...... is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing...
Directory of Open Access Journals (Sweden)
R. Rosario Diéguez Castro
2009-12-01
Full Text Available Se desarrolló y validó la técnica de Western Blot aplicada a la vacuna antimeningocócica Men B producida en el Instituto Finlay con el objetivo de demostrar un criterio de identidad. En el estudio de las proteínas antigénicas de la vacuna, P1.15 y P1.4 en vesícula de membrana externa,monograneles y producto final se emplearon en la identificación anticuerpos monoclonales específicos para estas proteínas. Los parámetros desarrollados en la validación de la técnica fueron: especificidad, límite de detección, repetibilidad, precisión intermedia, reproducibilidad y robustez. El método cumplió con los parámetros señalados, por lo que se consideró validado.Western Blot technique was developed and validated, applied to Men B meningococcal vaccine produced in "Carlos J, Finlay" Institute to demonstrate an identity criterion. In study of antigenic proteins of the vaccine, we used P1.15 y P1.4 in vesicle of external membrane, monogranels, and end product to identify the monoclonal antibodies specific of these proteins. Parameters developed in technique validation included: specificity, detection limit, repetition, average accuracy, reproduction, and strength. Method fulfilled with specified parameters, thus considering its validation.
Multiplexed Western Blotting Using Microchip Electrophoresis.
Jin, Shi; Furtaw, Michael D; Chen, Huaxian; Lamb, Don T; Ferguson, Stephen A; Arvin, Natalie E; Dawod, Mohamed; Kennedy, Robert T
2016-07-05
Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10-20 μg per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 μm deep × 50 μm wide × 8.6 cm long channel, it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g., ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng of total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps.
Western blotting using capillary electrophoresis.
Anderson, Gwendolyn J; M Cipolla, Cynthia; Kennedy, Robert T
2011-02-15
A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein adsorption. Although discrete protein zones could be detected, bands were broadened by ∼1.7-fold by transfer to membrane. A complete Western blot for lysozyme was completed in about one hour with 50 pg mass detection limit from low microgram per milliliter samples. These results demonstrate substantial reduction in time requirements and improvement in mass sensitivity compared to conventional Western blots. Western blotting using capillary electrophoresis shows promise to analyze low volume samples with reduced reagents and time, while retaining the information content of a typical Western blot.
Zinc blotting assay for detection of zinc binding prolamin in barley (Hordeum vulgare) grain
DEFF Research Database (Denmark)
Uddin, Mohammad Nasir; Nielsen, Ane Langkilde-Lauesen; Vincze, Eva
2014-01-01
In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol-soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. 65Zn overlay and blotting techniques have been widely used for detecting zinc......-binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (65ZnCl2) blotting technique was optimized to detect zinc-binding prolamins, followed by development of an easy-to-follow nonradioactive colorimetric...... zinc blotting method with a zinc-sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS-PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc-binding specificity of certain proteins was detected either by autoradiography or color...
OKABE, M; NYAKAS, C; BUWALDA, B; LUITEN, PGM
We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility
Fluorescence-based Western blotting for quantitation of protein biomarkers in clinical samples.
Zellner, Maria; Babeluk, Rita; Diestinger, Michael; Pirchegger, Petra; Skeledzic, Senada; Oehler, Rudolf
2008-09-01
Since most high throughput techniques used in biomarker discovery are very time and cost intensive, highly specific and quantitative analytical alternative application methods are needed for the routine analysis. Conventional Western blotting allows detection of specific proteins to the level of single isotypes while its quantitative accuracy is rather limited. We report a novel and improved quantitative Western blotting method. The use of fluorescently labelled secondary antibodies strongly extends the dynamic range of the quantitation and improves the correlation with the protein amount (r=0.997). By an additional fluorescent staining of all proteins immediately after their transfer to the blot membrane, it is possible to visualise simultaneously the antibody binding and the total protein profile. This allows for an accurate correction for protein load. Applying this normalisation it could be demonstrated that fluorescence-based Western blotting is able to reproduce a quantitative analysis of two specific proteins in blood platelet samples from 44 subjects with different diseases as initially conducted by 2D-DIGE. These results show that the proposed fluorescence-based Western blotting is an adequate application technique for biomarker quantitation and suggest possibilities of employment that go far beyond.
Western Blotting using Capillary Electrophoresis
Anderson, Gwendolyn J.; Cipolla, Cynthia; Kennedy, Robert T.
2011-01-01
A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein a...
Klaric , Laurent
2000-01-01
International audience; The unprecedented presence of Kostienki-technique prepared blades (also called Kostienki knives) in the Gravettian layers at Le Blot leads to a new analysis of these artefacts. Thorough technological study has pointed to the possible role of these items as cores, in association with or complementary to burin-forms, in particular context of backed-bladelet production. Le Blot is the second French site yielding such artefacts, the other being Corbiac (Dordogne). The aim ...
Brown, T
2001-05-01
Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support (the properties and advantages of the different types of membrane, transfer buffer, and transfer method are discussed in detail), resulting in immobilization of the DNA fragments, so the membrane carries a semipermanent reproduction of the banding pattern of the gel. After immobilization, the DNA can be subjected to hybridization analysis, enabling bands with sequence similarity to a labeled probe to be identified. This appendix describes Southern blotting via upward capillary transfer of DNA from an agarose gel onto a nylon or nitrocellulose membrane, using a high-salt transfer buffer to promote binding of DNA to the membrane. With the high-salt buffer, the DNA becomes bound to the membrane during transfer but not permanently immobilized. Immobilization is achieved by UV irradiation (for nylon) or baking (for nitrocellulose). A Support Protocol describes how to calibrate a UV transilluminator for optimal UV irradiation of a nylon membrane. An alternate protocol details transfer using nylon membranes and an alkaline buffer, and is primarily used with positively charged nylon membranes. The advantage of this combination is that no post-transfer immobilization step is required, as the positively charged membrane binds DNA irreversibly under alkaline transfer conditions. The method can also be used with neutral nylon membranes but less DNA will be retained. A second alternate protocol describes a transfer method based on a different transfer-stack setup. The traditional method of upward capillary transfer of DNA from gel to membrane described in the first basic and alternate protocols has certain disadvantages, notably the fact that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it. This slows down the blotting process and may reduce the amount of DNA that can be transferred. The downward capillary method described in
The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis
Silva, Jillian M.; McMahon, Martin
2014-01-01
The Western blot techniques that were originally established in the late 1970s are still actively utilized today. However, this traditional method of Western blotting has several drawbacks that include low quality resolution, spurious bands, decreased sensitivity, and poor protein integrity. Recent advances have drastically improved numerous aspects of the standard Western blot protocol to produce higher qualitative and quantitative data. The Bis-Tris gel system, an alternative to the convent...
Introduction to protein blotting.
Kurien, Biji T; Scofield, R Hal
2009-01-01
Protein blotting is a powerful and important procedure for the immunodetection of proteins following electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer.
Multistrip Western blotting: a tool for comparative quantitative analysis of multiple proteins.
Aksamitiene, Edita; Hoek, Jan B; Kiyatkin, Anatoly
2015-01-01
The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical Western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip Western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip Western blotting increases data output per single blotting cycle up to tenfold; allows concurrent measurement of up to nine different total and/or posttranslationally modified protein expression obtained from the same loading of the sample; and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data and therefore is advantageous to apply in biomedical diagnostics, systems biology, and cell signaling research.
Western blotting: an introduction.
Kurien, Biji T; Scofield, R Hal
2015-01-01
Western blotting is an important procedure for the immunodetection of proteins, particularly proteins that are of low abundance. This process involves the transfer of protein patterns from gel to microporous membrane. Electrophoretic as well as non-electrophoretic transfer of proteins to membranes was first described in 1979. Protein blotting has evolved greatly since the inception of this protocol, allowing protein transfer to be accomplished in a variety of ways.
Morinaga, Osamu; Uto, Takuhiro; Yuan, Chun-Su; Tanaka, Hiroyuki; Shoyama, Yukihiro
2010-06-01
A new eastern blotting technique has been established for ginsenoside Re (G-Re) contained in American ginseng berry pulp extracts. G-Re in American ginseng berry pulp was extracted using 100% methanol, 100% ethanol, 50% aqueous methanol, and 50% aqueous ethanol. The combined crude extracts were applied onto a polyethersulfone membrane and developed using the methanol-water-acetic acid solvent system (45:55:1 v/v). Separated components were immunostained using anti-G-Re monoclonal antibody. G-Re was first specifically detected and then quantitatively analyzed using NIH Imaging software. We also confirmed that the most suitable solvent was 50% aqueous methanol for extracting G-Re from American ginseng berry pulp. (c) 2009 Elsevier B.V. All rights reserved.
Multi-strip Western blotting to increase quantitative data output
Aksamitiene, Edita; Hoek, Jan B.; Kholodenko, Boris; Kiyatkin, Anatoly
2007-01-01
The qualitative and quantitative measurement of protein abundance and protein modification states are essential in understanding their role in diverse cellular processes. Traditional Western blotting technique, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. We propose a modified immunoblotting procedure, which is based on simultaneous transfer of proteins from multiple gel-strips onto the same membrane, and is compatible wi...
Detection of proteins on blot transfer membranes.
Sasse, Joachim; Gallagher, Sean R
2003-11-01
In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.
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Josefsen, Knud; Nielsen, Henrik
2011-01-01
is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing...
Kong, Ling-Quan; Pu, Ying-Hui; Ma, Shi-Kun
2008-01-01
To study how the choices of the quick vs slow protein transfer, the blotting membranes and the visualization methods influence the performance of Western blotting. The cellular proteins were abstracted from human breast cell line MDA-MB-231 for analysis with Western blotting using quick (2 h) and slow (overnight) protein transfer, different blotting membranes (nitrocellulose, PVDF and nylon membranes) and different visualization methods (ECL and DAB). In Western blotting with slow and quick protein transfer, the prestained marker presented more distinct bands on nitrocellulose membrane than on the nylon and PVDF membranes, and the latter also showed clear bands on the back of the membrane to very likely cause confusion, which did not occur with nitrocellulose membrane. PVDF membrane allowed slightly clearer visualization of the proteins with DAB method as compared with nitrocellulose and nylon membranes, and on the latter two membranes, quick protein transfer was likely to result in somehow irregular bands in comparison with slow protein transfer. With slow protein transfer and chemiluminescence for visualization, all the 3 membranes showed clear background, while with quick protein transfer, nylon membrane gave rise to obvious background noise but the other two membranes did not. Different membranes should be selected for immunoblotting according to the actual needs of the experiment. Slow transfer of the proteins onto the membranes often has better effect than quick transfer, and enhanced chemiluminescence is superior to DAB for protein visualization and allows highly specific and sensitive analysis of the protein expressions.
A streamlined Western blot exercise: An efficient and greener approach in the laboratory classroom.
Ness, Traci L; Robinson, Rebekah L; Mojadedi, Wais; Peavy, Lydia; Weiland, Mitch H
2015-01-01
SDS-PAGE and western blotting are two commonly taught protein detection techniques in biochemistry and molecular biology laboratory classrooms. A pitfall associated with incorporating these techniques into the laboratory is the significant wait times that do not allow students to obtain timely results. The waiting associated with SDS-PAGE comes from staining and destaining, whereas with western blotting it is the times required for antibody incubations and the numerous wash steps. This laboratory exercise incorporates 2,2,2-trichloroethanol (TCE) into the SDS-PAGE gel allowing for visualization of migrated proteins in a matter of minutes, saving both the time and chemical waste associated with traditional Coomassie staining. Additionally, TCE staining does not affect protein transfer eliminating the requirement for duplicated gels for total protein and western analyses. Protein transfer can be confirmed immediately without the use of Ponceau S staining. Lastly, this western blot procedure has been further shortened by using an HRP-conjugated primary antibody, which eliminates the secondary antibody incubation and washes, and uses a colorimetric detection to allow for visualization by students without the need for specialized equipment. © 2015 The International Union of Biochemistry and Molecular Biology.
HIV‑2 antibody detection after indeterminate or negative HIV‑1 Western blot in Cuba, 2005-2008.
Díaz, Dervel F; Ortiz, Eva; Martín, Dayamí; Nibot, Carmen; Rizo, Adis; Silva, Eladio
2012-01-01
that HIV-2 Western blot be included in the diagnostic algorithm for HIV-1/2 to followup negative or indeterminate HIV-1 Western blot results. KEYWORDS Diagnosis, laboratory techniques and procedures, antibodies, HIV-2, Western blot, enzyme-linked immunosorbent assay, algorithm, Cuba.
Solid-phase assay for the phosphorylation of proteins blotted on nitrocellulose membrane filters
International Nuclear Information System (INIS)
Valtorta, F.; Schiebler, W.; Jahn, R.; Ceccarelli, B.; Greengard, P.
1986-01-01
A new procedure for the phosphorylation and assay of phosphoproteins is described. Proteins are solubilized from tissue samples, separated by polyacrylamide gel electrophoresis, transferred onto nitrocellulose membrane filters, and the blotted polypeptides are phyosphorylated with the catalytic subunit of cyclic AMP (adenosine 3':5'-monophosphate)-dependent protein kinase. The method was developed for the assay of dephosphosynapsin I, but it has also proven suitable for the phosphorylation of other proteins. The patterns of phosphorylation of tissue samples phosphorylated using the new method are similar to those obtained using the conventional test tube assay. Once phosphorylated, the adsorbed proteins can be digested with proteases and subjected to phosphopeptide mapping. The phosphorylated blotted proteins can also be analyzed by overlay techniques for the immunological detection of polypeptides
Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis
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N.S. Sato
2004-07-01
Full Text Available Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples, from healthy blood donors (50 samples, individuals with sexually transmitted disease other than syphilis (3 samples, and from individuals with other spirochetal diseases such as Lyme disease (20 samples and leptospirosis (3 samples. The assay had sensitivity of 95.1% (95% CI, 86.1 to 98.7% and a specificity of 94.7% (95% CI, 87.0 to 98.7%; a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.
Directory of Open Access Journals (Sweden)
Yeda L. Nogueira
2007-12-01
Full Text Available The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.A técnica de Western blot foi utilizada para demonstrar a presença de anticorpos do soro de cães, que apresentavam leishmaniose visceral canina, contra algumas moléculas específicas no lisado da forma promastigota de Leshmania chagasi.Através da associa
Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation.
Rey, María-Dolores; Prieto, Pilar
2017-01-01
Simple, reliable methods for the identification of alien genetic introgressions are required in plant breeding programmes. The use of genomic dot-blot hybridisation allows the detection of small Hordeum chilense genomic introgressions in the descendants of genetic crosses between wheat and H. chilense addition or substitution lines in wheat when molecular markers are difficult to use. Based on genomic in situ hybridisation, DNA samples from wheat lines carrying putatively H. chilense introgressions were immobilised on a membrane, blocked with wheat genomic DNA and hybridised with biotin-labelled H. chilense genomic DNA as a probe. This dot-blot screening reduced the number of plants necessary to be analysed by molecular markers or in situ hybridisation, saving time and money. The technique was sensitive enough to detect a minimum of 5 ng of total genomic DNA immobilised on the membrane or about 1/420 dilution of H. chilense genomic DNA in the wheat background. The robustness of the technique was verified by in situ hybridisation. In addition, the detection of other wheat relative species such as Hordeum vulgare , Secale cereale and Agropyron cristatum in the wheat background was also reported .
Other notable protein blotting methods: a brief review.
Kurien, Biji T; Scofield, R Hal
2015-01-01
Proteins have been transferred from the gel to the membrane by a variety of methods. These include vacuum blotting, centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low- and high-molecular-weight proteins, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization mass spectrometric-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before matrix-assisted laser desorption/ionization tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.
Multistrip Western blotting to increase quantitative data output
Kiyatkin, Anatoly; Aksamitiene, Edita
2009-01-01
The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical Western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip Western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single mem...
Western Blot of Stained Proteins from Dried Polyacrylamide Gels
Gruber, Claudia; Stan-Lotter, Helga
1996-01-01
Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.
Post-staining electroblotting for efficient and reliable peptide blotting.
Lee, Der-Yen; Chang, Geen-Dong
2015-01-01
Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.
Genetic relatedness of orbiviruses by RNA-RNA blot hybridization
International Nuclear Information System (INIS)
Bodkin, D.K.
1985-01-01
RNA-RNA blot hybridization was developed in order to identify type-specific genes among double-stranded (ds) RNA viruses, to assess the genetic relatedness of dsRNA viruses and to classify new strains. Viral dsRNA segments were electrophoresed through 10% polyacrylamide gels, transferred to membranes, and hybridized to [5' 32 P]-pCp labeled genomic RNA from a related strain. Hybridization was performed at 52 0 C, 50% formamide, 5X SSC. Under these conditions heterologous RNA species must share ≥ 74% sequence homology in order to form stable dsRNA hybrids. Cognate genes of nine members of the Palyam serogroup of orbiviruses were identified and their sequence relatedness to the prototype. Palyam virus, was determined. Reciprocal blot hybridizations were performed using radiolabeled genomic RNA of all members of the Palyam serogroup. Unique and variant genes were identified by lack of cross-homology or by weak homology between segments. Since genes 2 and 6 exhibited the highest degree of sequence variability, response to the vertebrate immune system may be a major cause of sequence divergence among members of a single serogroup. Changuinola serogroup isolates were compared by dot-blot hybridization, while Colorado tick fever (CTF) serogroup isolates were compared by the RNA-RNA blot hybridization procedure described for reovirus and Palyam serogroup isolates. Preliminary blot hybridization data were also obtained on the relatedness of members of different Orbivirus serogroups
Inheritance of resistance to barley yellow dwarf virus detected by northern blot analysis
International Nuclear Information System (INIS)
Lorens, G.F.; Falk, B.W.; Qualset, C.O.
1989-01-01
Development of wheat (Triticum aestivum L.) cultivars tolerant to the barley yellow dwarf virus disease (BYD) has been limited by lack of precision in rating plants for response to infection, usually done by visual scoring of plant symptoms under field conditions. Other methodologies have been developed to study the host/pathogen relationship and to assess resistance or susceptibility. In this study northern dot blot analysis was used to determine barley yellow dwarf virus (BYDV) RNA concentrations of six wheat cultivars that differed in visual BYD symptom expression. Plants were infected with the NYPAV (PAV) isolate of BYDV in the greenhouse. At several dates after inoculation crude plant extracts were blotted on nitrocellulose and hybridized with a 32 P-labeled probe of the pPA8 cDNA clone of BYDV. The distribution of PRC for the F 2 population was compared to the distribution of BYD visual symptom scores for 403 F 2 plants of a similar F 2 population of NS 879/4 x Seri 82 under field conditions. The results were qualitatively similar, suggesting that northern dot blot analysis to measure PRC may be useful in understanding the genetics of resistance to BYD. This technique, when incorporated into breeding programs, could be important in the development of highly tolerant wheat cultivars with reduced losses to BYD
International Nuclear Information System (INIS)
Maria Lina R; Budiman Bela; Andi Yasmon
2009-01-01
Handling and controlling of tuberculosis, a disease caused by Mycobacterium tuberculosis (MTB), is now complicated since there are many MTBs that are resistant against anti-tuberculosis drugs such as isoniazid. The drug resistance could occurred due to the inadequate and un-regular drug utilization that cause gene mutation of the drug target such as katG gene for isoniazid. The molecular biology techniques such as the PCR- dot blot hybridization with radioisotope ( 32 P) labeled oligonucleotide probe, has been reported as a technique that is more sensitive and rapid for detection of gene mutations related with drug resistances. Hence, the aim of this study was to apply the PCR- dot blot hybridization technique using 32 P labeled oligonucleotide probe for detection of single mutation at codon 315 of katG gene of MTBs that rise the isoniazid resistance. In this study, we used 89 sputum specimens and a standard MTB (MTB H 37 RV) as a control. DNA extractions were performed by the BOOM method and the phenol chloroform for sputum samples and standard MTB, respectively. Primers used for PCR technique were Pt8 and Pt9 and RTB59 and RTB36 for detecting tuberculosis causing Mycobacterium and the existence of katG gene, respectively. Both of the primers are specific for IS6110 region and katG gene, respectively. PCR products were detected by an agarose gel electrophoresis technique. Dot blot hybridization with 32 P-oligonucleotide probe 315mu was performed to detect mutation at codon 315 of tested samples. Results of the PCR using primer Pt8 and Pt9 showed that all sputum specimens had positive results. Mutation detection by PCR- dot blot hybridization with 32 P-oligonucleotide probe 315mu, revealed that 11 of 89 tested samples had a mutation at their codon 315 of katG gene. Based upon these results, it is concluded that PCR-dot blot hybridization with 32 P-oligonucleotide probe is a technique that is rapid and highly specific and sensitive for detection of mutation at codon
Analysis of sperm antigens by sodium dodecyl sulfate gel/protein blot radioimmunobinding method
International Nuclear Information System (INIS)
Lee, C.Y.G.; Huang, Y.S.; Hu, P.C.; Gomel, V.; Menge, A.C.
1982-01-01
A radioimmunobinding method based on the blotting of renatured proteins from sodium dodecyl sulfate gels on to nitrocellulose filter papers was developed to analyze the sperm antigens that elicit serum anti-sperm antibodies. In rabbits, serum anti-sperm antibodies were raised by immunization with homologous epididymal spermatozoa mixed with complete Freund's adjuvant. The raised antisera from either male or female rabbits were shown to react with three major sperm protein bands on sodium dodecyl sulfate gels with the corresponding molecular weights of about 70,000 +/- 5000, 14,000, and 13,000, respectively. In humans, the monoclonal antibodies against human sperm were raised by a hybridoma technique. Out of six independent hybrid cell lines that were generated, three of them were shown to secrete immunoglobulins that react with the same two protein bands on sodium dodecyl sulfate gels, which have the approximate molecular weight of 10,000. The same procedure was also used to analyze human serum samples that were shown to contain anti-sperm antibodies by the known techniques. Unique sperm antigens that elicit anti-sperm antibodies in humans were identified and correlated. The results of this study suggest that sodium dodecyl sulfate gel/protein blot radioimmunobinding method may be a sensitive and useful tool for the study of sperm antigens that elicit autoimmune responses and their association with human infertility
Ultrasensitive Detection of Proteins on Western Blots with Semiconducting Polymer Dots
Ye, Fangmao; Smith, Polina B.; Wu, Changfeng; Chiu, Daniel T.
2013-01-01
We demonstrate ultrasensitive fluorescence imaging of proteins on Western blots using a bright, compact, and orange-emitting semiconducting polymer dot (CN-PPV). We achieved a detection limit at the single-picogram level in dot blots; with conventional Western blotting, we detected 50 pg of transferrin and trypsin inhibitor after SDS-PAGE and transfer onto a PVDF membrane. Our method does not require any additional equipment or time compared to the conventional procedure with traditional fluo...
A brief review of other notable protein blotting methods.
Kurien, Biji T; Scofield, R Hal
2009-01-01
A plethora of methods have been used for transferring proteins from the gel to the membrane. These include centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low and high molecular weight proteins, blotting of Coomassie Brilliant Blue (CBB)-stained proteins from polyacrylamide gels to transparencies, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before MALDI tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter.
An alternative method for processing northern blots after capillary transfer.
Nilsen, Timothy W
2015-03-02
Different laboratories use different methods for the prehybridization, hybridization, and washing steps of the northern blotting procedure. In this protocol, a northern blot is pretreated with Church and Gilbert hybridization buffer to block nonspecific probe-binding sites. The immobilized RNA is then hybridized to a DNA probe specific for the RNA of interest. Finally, the membrane is washed and subjected to autoradiography or phosphorimaging. The solutions and conditions described here may be ideal for those who prefer to use fewer ingredients in their solutions. This protocol is designed to achieve the same goals as other northern blotting approaches. It minimizes background (nonspecific adherence of probe to membrane and nonspecific hybridization) and maximizes specific hybridization to RNAs immobilized on a membrane. © 2015 Cold Spring Harbor Laboratory Press.
Directory of Open Access Journals (Sweden)
NUNES Cáris Maroni
1997-01-01
Full Text Available Visceral larva migrans (VLM is a clinical syndrome caused by infection of man by Toxocara spp, the common roundworm of dogs and cats. Tissue migration of larval stages causes illness specially in children. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. After the introduction of the enzyme-linked immunosorbent assay (ELISA using the larval excretory-secretory antigen of T. canis (TES, the diagnosis specificity was greatly improved although cross-reactivity with other helminths are still being reported. In Brazil, diagnosis is routinely made after absorption of serum samples with Ascaris suum antigens, a nematode antigenicaly related with Ascaris lumbricoides which is a common intestinal nematode of children. In order to identify T. canis antigens that cross react to A. suum antigens we analyzed TES antigen by SDS-PAGE and Western blotting techniques. When we used serum samples from patients suspected of VLM and positive result by ELISA as well as a reference serum sample numerous bands were seen (molecular weight of 210-200 kDa, 116-97 kDa, 55-50 kDa and 35-29 kDa. Among these there is at least one band with molecular weight around 55-66 kDa that seem to be responsible for the cross-reactivity between T. canis e A. suum once it disappears when previous absorption of serum samples with A. suum antigens is performed
SDS-Polyacrylamide Electrophoresis and Western Blotting Applied to the Study of Asthma.
García-Solaesa, Virginia; Abad, Sara Ciria
2016-01-01
Western blotting is used to analyze proteins after being separated by electrophoresis and subsequently electro-transferred to a membrane. Once immobilized, a specific protein can be identified through its reaction with a labeled antibody or antigen. It is a methodology commonly used in biomedical research such as asthma studies, to assess the pathways of inflammatory mediators involved in the disease.Here, we describe an example of western blotting to determine the factors involved in asthma. In this chapter, the methodology of western blotting is reviewed, paying attention on potential problems and giving interesting recommendations.
Energy Technology Data Exchange (ETDEWEB)
Kale, Sonia [Agharkar Research Institute (India); Kale, Anup [University of Alabama, Center for Materials for Information Technology (United States); Gholap, Haribhau; Rana, Abhimanyu [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Desai, Rama [National Centre for Cell Science (India); Banpurkar, Arun [University of Pune, Department of Physics (India); Ogale, Satishchandra, E-mail: sb.ogale@ncl.res.in [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Shastry, Padma, E-mail: padma@nccs.res.in [National Centre for Cell Science (India)
2012-03-15
In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and {beta} actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. In addition, these labels provide the capability of enhanced identification and localization of marker proteins in intact cells by confocal laser scanning microscopy.
A Laboratory Exercise Illustrating the Sensitivity and Specificity of Western Blot Analysis
Chang, Ming-Mei; Lovett, Janice
2011-01-01
Western blot analysis, commonly known as "Western blotting," is a standard tool in every laboratory where proteins are analyzed. It involves the separation of polypeptides in polyacrylamide gels followed by the electrophoretic transfer of the separated polypeptides onto a nitrocellulose or polyvinylidene fluoride membrane. A replica of the…
Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting.
Heda, Ghanshyam D; Omotola, Oluwabukola B; Heda, Rajiv P; Avery, Jamie
2016-09-01
SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin's transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting.
Shiokai, Sachiko; Kitashiba, Hiroyasu; Nishio, Takeshi
2010-08-01
Although the dot-blot-SNP technique is a simple cost-saving technique suitable for genotyping of many plant individuals, optimization of hybridization and washing conditions for each SNP marker requires much time and labor. For prediction of the optimum hybridization conditions for each probe, we compared T (m) values estimated from nucleotide sequences using the DINAMelt web server, measured T (m) values, and hybridization conditions yielding allele-specific signals. The estimated T (m) values were comparable to the measured T (m) values with small differences of less than 3 degrees C for most of the probes. There were differences of approximately 14 degrees C between the specific signal detection conditions and estimated T (m) values. Change of one level of SSC concentrations of 0.1, 0.2, 0.5, and 1.0x SSC corresponded to a difference of approximately 5 degrees C in optimum signal detection temperature. Increasing the sensitivity of signal detection by shortening the exposure time to X-ray film changed the optimum hybridization condition for specific signal detection. Addition of competitive oligonucleotides to the hybridization mixture increased the suitable hybridization conditions by 1.8. Based on these results, optimum hybridization conditions for newly produced dot-blot-SNP markers will become predictable.
Pazos, Raquel; Echevarria, Juan; Hernandez, Alvaro; Reichardt, Niels-Christian
2017-09-01
Aberrant protein glycosylation is a hallmark of cancer, infectious diseases, and autoimmune or neurodegenerative disorders. Unlocking the potential of glycans as disease markers will require rapid and unbiased glycoproteomics methods for glycan biomarker discovery. The present method is a facile and rapid protocol for qualitative analysis of protein glycosylation in complex biological mixtures. While traditional lectin arrays only provide an average signal for the glycans in the mixture, which is usually dominated by the most abundant proteins, our method provides individual lectin binding profiles for all proteins separated in the gel electrophoresis step. Proteins do not have to be excised from the gel for subsequent analysis via the lectin array but are transferred by contact diffusion from the gel to a glass slide presenting multiple copies of printed lectin arrays. Fluorescently marked glycoproteins are trapped by the printed lectins via specific carbohydrate-lectin interactions and after a washing step their binding profile with up to 20 lectin probes is analyzed with a fluorescent scanner. The method produces the equivalent of 20 lectin blots in a single experiment, giving detailed insight into the binding epitopes present in the fractionated proteins. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.
Direct tissue blot immunoassay for detection of Xylella fastidiosa in olive trees
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Khaled DJELOUAH
2015-01-01
Full Text Available A direct tissue blot immunoassay (DTBIA technique has been compared with ELISA and PCR for detection of Xylella fastidiosa in olive trees from Apulia (southern Italy. Fresh cross-sections of young twigs and leaf petioles were printed onto nitrocellulose membranes and analyzed in the laboratory. Analyses of a first group of 61 samples gave similar efficiency for the three diagnostic techniques for detection the bacterium (24 positive and 36 negative samples, except for a single sample which was positive only with DTBIA and PCR. Similar results were obtained by separately analyzing suckers and twigs collected from different sectors of tree canopies of a second group of 20 olive trees (ten symptomatic and ten symptomless. In this second test the three diagnostic techniques confirmed the irregular distribution of the bacterium in the tree canopies and erratic detectability of the pathogen in the young suckers. It is therefore necessary to analyse composite samples per tree which should be prepared with twigs collected from different sides of the canopy. The efficiency comparable to ELISA and PCR, combined with the advantages of easier handling, speed and cost, make DTBIA a valid alternative to ELISA in large-scale surveys for occurrence of X. fastidiosa. Moreover, the printing of membranes directly in the field prevents infections spreading to Xylella-free areas, through movement of plant material with pathogen vectors for laboratory testing.
Positive IgG Western Blot for Borrelia burgdorferi in Colombia
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Palacios Ricardo
1999-01-01
Full Text Available In order to evaluate the presence of specific IgG antibodies to Borrelia burgdorferi in patients with clinical manifestations associated with Lyme borreliosis in Cali, Colombia, 20 serum samples from patients with dermatologic signs, one cerebrospinal fluid (CSF sample from a patient with chronic neurologic and arthritic manifestations, and twelve serum samples from individuals without clinical signs associated with Lyme borreliosis were analyzed by IgG Western blot. The results were interpreted following the recommendations of the Centers for Diseases Control and Prevention (CDC for IgG Western blots. Four samples fulfilled the CDC criteria: two serum specimens from patients with morphea (localized scleroderma, the CSF from the patient with neurologic and arthritic manifestations, and one of the controls. Interpretation of positive serology for Lyme disease in non-endemic countries must be cautious. However these results suggest that the putative "Lyme-like" disease may correlate with positivity on Western blots, thus raising the possibility that a spirochete genospecies distinct from B. burgdorferi sensu stricto, or a Borrelia species other than B. burgdorferi sensu lato is the causative agent. Future work will focus on a survey of the local tick and rodent population for evidence of spirochete species that could be incriminated as the etiologic agent.
Khanna, Rahul; Rao, G R K; Tiwary, S K; Rai, Ashish; Khanna, Seema; Khanna, A K
2009-04-01
The etiopathological role of human papilloma virus (HPV) in the causation of oral cancer is till a subject of speculation. We used the technique of Southern blot hybridization to detect the presence of HPV types 16 & 18 in biopsy specimens from oral cancer and leukoplakia patients as well as normal oral mucosal biopsies. The prevalence of either HPV type 16 or 18 was found in 64.5% (29/45) of oral cancer, 40%(12/30) of leukoplakia and 20%(9/45) of normal oral mucosal biopsies. No association could be demonstrated between tobacco usage habits or a history of genital warts with HPV prevalence. A significant finding was that none of the oral cancer patients were negative for both: a history of tobacco usage as well as presence of HPV infection, on Southern blot hybridization.
BLOTS AND ALL: A HISTORY OF THE RORSCHACH INK BLOT TEST IN BRITAIN.
Hubbard, Katherine; Hegarty, Peter
2016-01-01
Despite the easily recognizable nature of the Rorschach ink blot test very little is known about the history of the test in Britain. We attend to the oft-ignored history of the Rorschach test in Britain and compare it to its history in the US. Prior to the Second World War, Rorschach testing in Britain had attracted advocates and critiques. Afterward, the British Rorschach Forum, a network with a high proportion of women, developed around the Tavistock Institute in London and The Rorschach Newsletter. In 1968, the International Rorschach Congress was held in London but soon after the group became less exclusive, and fell into decline. A comparative account of the Rorschach in Britain demonstrates how different national institutions invested in the 'projective hypothesis' according to the influence of psychoanalysis, the adoption of a nationalized health system, and the social positioning of 'others' throughout the twentieth century. In comparing and contrasting the history of the Rorschach in Britain and the US, we decentralize and particularize the history of North American Psychology. © 2016 Wiley Periodicals, Inc.
Davelois, Kelly; Escalante, Hermes; Jara, César
2016-01-01
. To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.
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Neuza Satomi SATO
1999-03-01
Full Text Available Three GST fusion recombinant antigen of Treponema pallidum, described as GST-rTp47, GST-rTp17 and GST-rTp15 were analyzed by Western blotting techniques. We have tested 53 serum samples: 25 from patients at different clinical stages of syphilis, all of them presenting anti-treponemal antibody, 25 from healthy blood donors and three from patients with sexually transmitted disease (STD other than syphilis. Almost all samples from patients with syphilis presented a strong reactivity with GST-rTp17 antigen. Some samples were non-reactive or showed a weak reaction with GST-rTp47 and/or GST-rTp15, and apparently there was no correlation with the stage of disease. There was no seropositivity among blood donors. No sample reacted with purified GST. We concluded that due to their specificity these recombinant antigens can be used as GST fusion protein for development of syphilis diagnostic assays.Os antígenos recombinantes de Treponema pallidum GST-rTp47, GST-rTp17 e GST-rTp15, produzidos em fusão com glutationa S-transferase (GST em E. coli, foram analisados quanto ao potencial diagnóstico da sífilis pela técnica de Western blotting. Foram testadas 53 amostras, sendo 25 de pacientes em diferentes estágios clínicos da sífilis, com resultados positivos no teste treponêmico clássico; 25 amostras procedentes de doadores de banco de sangue, com sorologia negativa e 3 de pacientes com doença sexualmente transmissível não relacionado à sífilis. Todas as amostras de pacientes com sífilis apresentaram alta reatividade com o antígeno GST-rTp17. Quanto aos antígenos GST-rTp47 e GST-Tp15 verificou-se uma variação na presença ou na intensidade da reação em diferentes amostras de pacientes com sífilis, sem mostrar correlação com o estágio da doença. Nenhuma reatividade contra quaisquer desses antígenos foi observada com as amostras do grupo controle. Nenhuma das amostras testadas apresentaram reatividade com a GST purificada. A
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Hermes Escalante
2011-09-01
finding of parasite eggs in the stool microscopy. Antigens of 10, 12, 17, 23, 27, 30, 36, 43, 66 and 136 kDa were detected and used to develop the Western blot technique. The sensitivity was evaluated using sera from 67 fasciolosis patients, and the specificity using sera from 57 patients with other parasitic diseases, and 10 from healthy individuals. Results. Out of the 67 sera, 64 reacted with the 23 kDa band and 61 with the one of 17 kDa. These two bands were not detected in sera from patients with other parasitic diseases or in those from healthy volunteers and thus could be considered specific and diagnostic. Conclusions. The sensitivity of the test, using the bands of 17 and 23 kDa, was 95.5% for positive reactions to at least one of these two bands, being its specificity 100% with a positive predictive value of 100% and negative predictive value of 95.71%.
Blotting from PhastGel to Membranes by Ultrasound.
Kost, Joseph; Azagury, Aharon
2015-01-01
Ultrasound based approach for enhanced protein blotting is proposed. Three minutes of ultrasound exposure (1 MHz, 2.5 W/cm(2)) was sufficient for a clear transfer of proteins from a polyacrylamide gel (PhastGel) to nitrocellulose or Nylon 66 Biotrans membrane. The proteins evaluated were prestained sodium dodecyl sulfate-polyacrylamide standards (18,500-106,000 Da) and 14C-labeled Rainbow protein molecular weight markers (14,300-200,000 Da).
Rossano, M G; Mansfield, L S; Kaneene, J B; Murphy, A J; Brown, C M; Schott, H C; Fox, J C
2000-01-01
Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by the apicomplexan protozoan parasite Sarcocystis neurona. The purposes of this study were to develop the most stringent criteria possible for a positive test result, to estimate the sensitivity and specificity of the EPM Western blot antibody test, and to assess the ability of bovine antibodies to Sarcocystis cruzi to act as a blocking agent to minimize false-positive results in the western blot test for S. neurona. Sarcocystis neurona merozoites harvested from equine dermal cell culture were heat denatured, and the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in a 12-20% linear gradient gel. Separated proteins were electrophoretically transferred to polyvinylidene fluoride membranes and blocked in 1% bovine serum albumin and 0.5% Tween-Tris-buffered saline. Serum samples from 6 horses with S. neurona infections (confirmed by culture from neural tissue) and 57 horses without infections (horses from the Eastern Hemisphere, where S. neurona does not exist) were tested by Western blot. Horses from both groups had reactivity to the 62-, 30-, 16-, 13-, 11-, 10.5-, and 10-kD bands. Testing was repeated with another step. Blots were treated with bovine S. cruzi antibodies prior to loading the equine samples. After this modification of the Western blot test, positive infection status was significantly associated with reactivity to the 30- and 16-kD bands (Pblot had a sample sensitivity of 100% and sample specificity of 98%. It is concluded that the specificity of the Western blot test is improved by blocking proteins not specific to S. neurona and using reactivity to the 30- and 16-kD bands as the criterion for a positive test.
Abeyrathne, Priyanka D; Lam, Joseph S
2007-04-01
A major hurdle in characterizing bacterial membrane proteins by Western blotting is the ineffectiveness of transferring these proteins from sodium dodecyl sulfate -- polyacrylamide gel electrophoresis (SDS-PAGE) gel onto nitrocellulose membrane, using standard Western blot buffers and electrophoretic conditions. In this study, we compared a number of modified Western blotting buffers and arrived at a composition designated as the SDS-PAGE-Urea Lysis buffer. The use of this buffer and specific conditions allowed the reproducible transfer of highly hydrophobic bacterial membrane proteins with 2-12 transmembrane-spanning segments as well as soluble proteins onto nitrocellulose membranes. This method should be broadly applicable for immunochemical studies of other membrane proteins.
Banding pattern indicative of echinococcosis in a commercial cysticercosis western blot
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Tappe D
2009-09-01
Full Text Available Abstract Objective A commercial cysticercosis Western blot was evaluated for serological cross-reactivity of sera from patients with alveolar (AE and cystic echinococcosis (CE. Methods A total of 161 sera were examined, including 31 sera from AE-patients, 11 sera from CE-patients, 9 sera from patients with other parasitic diseases and 109 sera from patients with unrelated medical conditions. All AE-and CE-sera were also examined by the echinococcosis Western blot. Results More sera from patients with AE than with CE showed cross-reactivity in the form of ladder-like patterns ("Mikado aspect" and untypical bands at 6-8 kDa (71% and 77.4% versus 27.3% and 45.5%, respectively. In contrast, triplets of bands in the area above 50 kDa and between 24 and 39-42 kDa were more frequent in CE than in AE sera. The fuzzy band at 50-55 kDa typical for cysticercosis was absent in all AE and CE sera. Conclusions Atypical banding patterns in the cysticercosis Western blot should raise the suspicion of a metacestode infection different from Taenia solium, i.e. Echinococcus multilocularis or E. granulosus, especially when the Mikado aspect and an altered 6-8 kDa band is visible in the absence of a fuzzy 50-55 kDa band.
Characterization of the structure of the erythropoietin receptor by ligand blotting
International Nuclear Information System (INIS)
Atkins, H.L.; Broudy, V.C.; Papayannopoulou, T.
1991-01-01
Erythropoietin (Epo) regulates the growth and differentiation of erythroid cells by binding to a specific receptor. We characterized the native Epo receptor on erythroleukemia cell lines by ligand blotting. Solubilized cell membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred onto nitrocellulose, and probed with 125I-Epo. Specificity was demonstrated by inhibition of 125I-Epo binding by unlabeled excess Epo but not other peptide growth factors and by the cellular distribution of the Epo binding protein. A single membrane protein of 61 Kd ± 4 Kd was sufficient to bind 125I Epo in both human (OCIM2, K562) and murine (GM979, Rauscher, DA-1) cell lines. This finding is consistent with the predicted size of the Epo receptor from the murine cDNA clone. However, chemical crosslinking of 125I-Epo to its receptor has identified two Epo binding proteins of 105 Kd and 85 Kd. This difference may occur because the receptor is size fractionated before Epo binding in the ligand blot, but after Epo binding in crosslinking studies. Ligand blotting demonstrates that the native Epo receptor is composed of a single 61-Kd Epo binding protein, and suggests the presence of additional proteins of 20 to 25 Kd that associate with the receptor after Epo binding
Standardization of Licorice and TCM Formulations Using Eastern Blot Fingerprinting Analysis
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Yukihiro Shoyama
2013-01-01
Full Text Available To prepare the antiglycyrrhizin (GC monoclonal antibody (MAb, GC was treated with NaIO4 resulting in aldehyde which can be combined with carrier protein. An antigen conjugate was performed by a matrix-assisted laser desorption/ionization TOF mass spectrometry to determine the hapten numbers in the conjugate. Anti-GC MAb was prepared from a hybridoma which was fixed from the spleen cells producing anti-GC MAb and the myeloma cells after immunization. The TCM and licorice extract were developed by TLC and blotted to a polyvinylidene difluoride (PVDF membrane. The membrane was treated by NaIO4 and protein, enzyme labeled secondary MAb, and finally substrate was added. Clear spot appeared on PVDF membrane identifying GC against a background containing large amount of impurities. In eastern blotting, the GC molecule was divided into two functions. The aglycone part is recognized as an epitope and the sugar moiety can be combined to membrane. The specific reactivity of sugar moiety in the GC molecule against anti-GC MAb might be modified by the NaIO4 treatment on the membrane because glycyrrhetic acid 3-O-glucuronide can be stained although the cross-reactivity is only 4.3%. Eastern blotting for GC can not only apply for the standardization of licorice and TCM, but also it can open for the other bioactive products.
Routine Western blot to check autophagic flux : Cautions and recommendations
Gomez-Sanchez, Ruben; Pizarro-Estrella, Elisa; Yakhine-Diop, Sokhna M. S.; Rodriguez-Arribas, Mario; Bravo-San Pedro, Jose M.; Fuentes, Jose M.; Gonzalez-Polo, Rosa A.
2015-01-01
At present, the analysis of autophagic flux by Western blotting (WB), which measures two of the most important markers of autophagy, i.e., microtubule-associated protein 1 light chain 3 (LC3) and p62, is widely accepted in the scientific community. In this study, we addressed the possible
DEFF Research Database (Denmark)
Christensen, C B; Theander, T G
1997-01-01
Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20% of the ......Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20......% of the PBMC cultures whereas proliferation was induced in 79% of the same cultures offered similar treated TT (except for the PAGE separation). When T-cell blotting was performed with TT separated in a SDS-agarose matrix, proliferation was induced in 80% of donors responding to soluble TT. The results show...... that SDS-PAGE alters the ability of TT to induce T-cell proliferation, possibly due to unpolymerized acrylamide binding to proteins during SDS-PAGE. The use of SDS-PAGE T-cell blotting in the screening for T-cell antigens must therefore be reconsidered. We suggest the use of SDS-Agarose Gel Electrophoresis...
Banks, Rosamonde E; Craven, Rachel A; Harnden, Patricia A; Selby, Peter J
2003-04-01
Western blotting remains a central technique in confirming identities of proteins, their quantitation and analysis of various isoforms. The biotin-avidin/streptavidin system is often used as an amplification step to increase sensitivity but in some tissues such as kidney, "nonspecific" interactions may be a problem due to high levels of endogenous biotin-containing proteins. The EnVision system, developed for immunohistochemical applications, relies on binding of a polymeric conjugate consisting of up to 100 peroxidase molecules and 20 secondary antibody molecules linked directly to an activated dextran backbone, to the primary antibody. This study demonstrates that it is also a viable and sensitive alternative detection system in Western blotting applications.
Ołdak, Elzbieta; Rozkiewicz, Doroto; Sulik, Artur
2008-01-01
In the afforested area of North-Eastern Poland the risk of Borrelia burgdorferi infection seems to be higher compared to the other regions. Because of unspecific clinical manifestation of Lyme borreliosis in children the positive ELISA IgM results should be confirmed with Western blot IgM tests. Retrospective analysis of clinical signs and symptoms of Lyme borreliosis in children with positive ELISA IgM and positive Western blot IgM results and in children with positive ELISA IgM and negative Western blot IgM results. The study included 20 children reactive with ELISA IgM (Bellco Biomedica, Austria), hospitalized in Pediatric Infectious Diseases Clinic in 2007 due to probable diagnosis of Lyme disease. All children were tested with B. burgdorferi Western blot IgM and/or IgG assay (DRG, Diagnostics, Germany) as a second-step diagnosis. In 10 (50% females, 50% males) out of 20 children the results were positive (borreliosis) and in other 10 (80% females, 20% males) the results were negative (controls). In both groups of patients the retrospective analysis of signs and symptoms was done. The most often clinical manifestation of Lyme borreliosis in children was neuroborreliosis. Children presented Lyme meningitis (30%), facial nerve palsy (10%) and chronic or recurrent headaches (40%), associated with vertigo (20%), weakness (30%), fever (40%), and fatigue syndrome (30%). One patient presented Lyme arthritis. Children of control group presented with unspecific symptoms like isolated headaches (40%), arthralgias (70%), myalgias (10%) and abdomen pain (20%) (1) The most frequent clinical presentation of Lyme borreliosis in analyzed children was neuroborreliosis; (2) Isolated arthralgias in children reactive with B. burgdorferi ELISA IgM need to be confirmed with Western blot assay before implementing the antibiotic therapy.
Total protein analysis as a reliable loading control for quantitative fluorescent Western blotting.
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Samantha L Eaton
Full Text Available Western blotting has been a key technique for determining the relative expression of proteins within complex biological samples since the first publications in 1979. Recent developments in sensitive fluorescent labels, with truly quantifiable linear ranges and greater limits of detection, have allowed biologists to probe tissue specific pathways and processes with higher resolution than ever before. However, the application of quantitative Western blotting (QWB to a range of healthy tissues and those from degenerative models has highlighted a problem with significant consequences for quantitative protein analysis: how can researchers conduct comparative expression analyses when many of the commonly used reference proteins (e.g. loading controls are differentially expressed? Here we demonstrate that common controls, including actin and tubulin, are differentially expressed in tissues from a wide range of animal models of neurodegeneration. We highlight the prevalence of such alterations through examination of published "-omics" data, and demonstrate similar responses in sensitive QWB experiments. For example, QWB analysis of spinal cord from a murine model of Spinal Muscular Atrophy using an Odyssey scanner revealed that beta-actin expression was decreased by 19.3±2% compared to healthy littermate controls. Thus, normalising QWB data to β-actin in these circumstances could result in 'skewing' of all data by ∼20%. We further demonstrate that differential expression of commonly used loading controls was not restricted to the nervous system, but was also detectable across multiple tissues, including bone, fat and internal organs. Moreover, expression of these "control" proteins was not consistent between different portions of the same tissue, highlighting the importance of careful and consistent tissue sampling for QWB experiments. Finally, having illustrated the problem of selecting appropriate single protein loading controls, we demonstrate
Binnicker, M J; Jespersen, D J; Harring, J A; Rollins, L O; Bryant, S C; Beito, E M
2008-07-01
The diagnosis of Lyme borreliosis (LB) is commonly made by serologic testing with Western blot (WB) analysis serving as an important supplemental assay. Although specific, the interpretation of WBs for diagnosis of LB (i.e., Lyme WBs) is subjective, with considerable variability in results. In addition, the processing, reading, and interpretation of Lyme WBs are laborious and time-consuming procedures. With the need for rapid processing and more objective interpretation of Lyme WBs, we evaluated the performances of two automated interpretive systems, TrinBlot/BLOTrix (Trinity Biotech, Carlsbad, CA) and BeeBlot/ViraScan (Viramed Biotech AG, Munich, Germany), using 518 serum specimens submitted to our laboratory for Lyme WB analysis. The results of routine testing with visual interpretation were compared to those obtained by BLOTrix analysis of MarBlot immunoglobulin M (IgM) and IgG and by ViraScan analysis of ViraBlot and ViraStripe IgM and IgG assays. BLOTrix analysis demonstrated an agreement of 84.7% for IgM and 87.3% for IgG compared to visual reading and interpretation. ViraScan analysis of the ViraBlot assays demonstrated agreements of 85.7% for IgM and 94.2% for IgG, while ViraScan analysis of the ViraStripe IgM and IgG assays showed agreements of 87.1 and 93.1%, respectively. Testing by the automated systems yielded an average time savings of 64 min/run compared to processing, reading, and interpretation by our current procedure. Our findings demonstrated that automated processing and interpretive systems yield results comparable to those of visual interpretation, while reducing the subjectivity and time required for Lyme WB analysis.
International Nuclear Information System (INIS)
Semenkovich, C.F.; Ostlund, R.E. Jr.; Yang, J.; Reaban, M.E.
1985-01-01
We report the detection of low-density lipoprotein (LDL) receptors by the technique of receptor blotting in fibroblasts from a patient with homozygous familial hypercholesterolemia (FHC) previously classified as ''receptor negative.'' Solubilized receptors were electrophoresed, transferred to nitrocellulose paper, treated with LDL followed by radiolabeled antibody to LDL, and visualized by autoradiography. GM 2000 FHC fibroblasts revealed LDL receptors with an apparent molecular weight of approximately 140,000, the same as in normal cells. LDL receptor activity by blotting in GM 2000 cells was greatly diminished in comparison with normal cells, but was calcium dependent. Receptor activity was also detectable by conventional monolayer binding and degradation assays. Thus, GM 2000 cells have profoundly diminished LDL receptor activity, but retain the genetic capacity to make LDL receptor material of normal molecular weight that is capable of binding LDL. Previous studies have demonstrated the presence of trace amounts of immunoreactive LDL receptor protein in fibroblasts from some receptor-negative FHC homozygotes. These studies are extended by demonstrating the ability of this material to bind LDL
Khrouf, Fatma; M'Ghirbi, Youmna; Znazen, Abir; Ben Jemaa, Mounir; Hammami, Adnene
2014-01-01
Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia. PMID:24226919
COMPARISONS OF ELISA AND WESTERN BLOT ASSAYS FOR DETECTION OF CRYPTOSPORIDIUM ANTIBODY
A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurre...
International Nuclear Information System (INIS)
Kroczek, R.A.; Siebert, E.
1990-01-01
We have optimized Northern analysis at several steps. Overnight electrophoresis was replaced by short gel runs and overnight capillary transfer by rapid vacuum-blotting adapted to Northern analysis. Short uv irradiation was used as a substitute for the usual RNA fixation by baking. Direct staining of RNA before electrophoresis made it possible to check RNA integrity and to evaluate the quality of the size separation immediately after electrophoresis. In this system, RNA transfer onto the membrane support could also be quickly assessed after the blotting step. The net result of all modifications was a doubling of the autoradiography signal compared with that obtained by modern Northern protocols. At the same time, the duration of the procedure was shortened drastically, allowing an autoradiography signal to be obtained within 24 h
Determination of Diagnostic Antigens in Cattle Amphistomiasis Using Western Blotting
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A Halajian
2009-05-01
Full Text Available "nBackground: Mixed infection with amphistomes seems common in native cattle of Iran. The aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis."nMethods: Specific antigens of Cotylophoron cotylophorum, Gastrothylax crumenifer and Paramphistomum cervi (mixed infection, the most common species, were collected from cattle was determined. Adult trematodes were collected from the rumen of naturally infected cattle at meat inspection. After their homogenization and centrifugation, somatic antigens were prepared and analyzed by SDS-PAGE. Specific antigens were determinated by western blot with homologous and heterologous sera. SDS-PAGE of whole worms extract was performed at different concentrations and subsequent gels staining. Immunoblotting analysis using sera from cattle naturally infected with amphistomes, Dicrocoelium dendriticum, Fasciola spp. and hydatid cyst was performed."nResults: Electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 µgr/ml and 8 µgr/ml with molecular weights ranging from 25-120 and 25-150 kDa, respectively. The best result was taken at 8 mg/ml concentration. Although western blot of these proteins demonstrate 5 major antigenic polypeptides ranging from 50 to 100 kDa which were recognized by serum of cattle naturally infected with mixed amphistomes.
Silver and gold nanoparticle coated membranes applied to protein dot blots
International Nuclear Information System (INIS)
Xie, F.; Drozdowicz-Tomsia, K.; Shtoyko, T.; Goldys, E. M.
2011-01-01
Detection and identification of low abundance biomarker proteins is frequently based on various types of membrane-based devices. Lowering of the protein detection limits is vital in commercial applications such as lateral flow assays and in Western blots widely used in proteomics. These currently suffer from insufficient detection sensitivity and low retention for small 2–5 kDa proteins. In this study, we report the deposition of two types of metal nanoparticles: gold colloids (50–95 nm diameter) and silver fractals onto a range of commonly used types of membranes including polyvinylidene fluoride (PVDF). Due to strong affinity of proteins to noble metals, such modified membranes have the potential to effectively capture trace proteins preventing their loss. The membranes modified by metal particles were characterized optically and by SEM. The membrane performance in protein dot blots was evaluated using the protein—fluorophore conjugates Deep Purple-bovine serum albumin and fluorescein—human serum albumin. We found that the metal nanoparticles increase light extinction by metals, which is balanced by increased fluorescence, so that the effective fluorescence signal is unchanged. This feature combined with the capture of proteins by the nanoparticles embedded in the membrane increases the detection limit of membrane assays.
Northern and Southern blot analysis of human RNA and DNA in autopsy material
DEFF Research Database (Denmark)
Larsen, S; Rygaard, K; Asnaes, S
1992-01-01
was obtained less than two days postmortem. Histological examination showing slight or no autolysis and the presence of ribosomal bands after gel electrophoresis were both indicative parameters of RNA preservation. DNA was appropriate for Southern blotting when the tissue was obtained less than three to five...
Checking transfer efficiency and equal loading via qualitative optical way in western blotting.
Gong, Jun-Hua; Gong, Jian-Ping; Zheng, Kai-Wen
2017-11-01
The ability to determine that successful transfer and equal loading occur prior to using primary antibodies is important. And total protein staining is commonly used to check transfer efficiency and normalization, which play a crucial role in western blotting. Ponceau S and coomassie blue are commonly used, but there are disadvantages reported in recent years. Therefore, we are interested in finding another method, which is cheap, easy and fast. As we know, protein binding region of PVDF membrane is still hydrophilic when carbinol volatilizes, however, the non-protein binding region of PVDF membrane became hydrophobic again. And this different wettability between non-protein binding region and protein binding region of Polyvinylidene difluoride membrane may be used to check transfer efficiency and equal loading in western blotting. Based on the principle above, we describe an optical approach where an experimenter can observe that the proteins have been transferred to the membrane without any staining within minutes. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Identification of toxocara canis antigens by Western blot in experimentally infected rabbits
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MORALES Olga Lucía
2002-01-01
Full Text Available Toxocariasis is a frequent helminthiasis that can cause visceral and ocular damage in humans specially in children. The identification of specific antigens of Toxocara canis is important in order to develop better diagnostic techniques. Ten rabbits were infected orally with a dose of 5000 Toxocara canis embryonated eggs. Rabbits were bled periodically and an ELISA assay was performed to determine levels of specific Toxocara IgG antibodies. ELISA detected antibodies at day 15 after infection. Western blot (WB assay was performed using excretory/secretory antigens (E/S of T. canis second stage larvae. Different antigen concentrations were evaluated: 150, 200, 250 and 300 µg/mL. The concentration of 250 µg/mL was retained for analysis. Rabbit sera were diluted 1:100. Secondary antibody was used at a dilution of 1:1000. Results of WB indicated that in the first month after infection specific antibodies against the 200 KDa, 116 KDa, 92 KDa and 35 KDa antigens were detected; antibodies against the 92 KDa, 80 KDa, 66 KDa, 45 KDa, 31 KDa and 28 KDa antigens appeared later. All positive sera in the ELISA test were also positive in WB. Two antigen bands, 92 KDa and 35 KDa, were identified since the beginning and throughout the course of infection. These antigens merit further evaluation as candidates for use in diagnosis.
Khammari, Imen; Saghrouni, Fatma; Yaacoub, Alia; Gaied Meksi, Sondoss; Ach, Hinda; Garma, Lamia; Fathallah, Akila; Ben Saïd, Moncef
2013-08-01
The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.
Acid-Urea Gel Electrophoresis and Western Blotting of Histones.
Hazzalin, Catherine A; Mahadevan, Louis C
2017-01-01
Acid-urea gel electrophoresis offers significant advantages over SDS-PAGE for analysis of post-translational protein modifications, being capable of resolving proteins of similar size but varying in charge. Hence, it can be used to separate protein variants with small charge-altering differences in primary sequence, and is particularly useful in the analysis of histones whose charge variation arises from post-translational modification, such as phosphorylation or acetylation. On acid-urea gels, histones that carry multiple modifications, each with a characteristic charge, are resolved into distinct bands, the so-called "histone ladder." Thus, the extent and distribution of different modification states of histones can be visualized. Here, we describe the analysis of histone H3 by acid-urea gel electrophoresis and western blotting.
Maccari, Francesca; Volpi, Nicola
2002-09-01
We describe a method for blotting and immobilizing several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by a cationic detergent after their separation by conventional agarose gel electrophoresis. Nitrocellulose membranes were derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of glycosaminoglycans (GAGs) were capillary-blotted after their separation in agarose gel electrophoresis in barium acetate/1,2-diaminopropane. Single purified species of variously sulfated polysaccharides were transferred onto the derivatized membranes after electrophoresis with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining) permitting about 0.1 nug threshold of detection. Nonsulfated polyanions, hyaluronic acid, a fructose-containing polysaccharide with a chondroitin backbone purified from Escherichia coli U1-41, and its defructosylated product, were also electrophoretically separated and transferred onto membranes. The limit of detection for desulfated GAGs was about 0.1-0.5 nug after irreversible or reversible staining. GAG extracts from bovine, lung and aorta, and human aorta and urine were separated by agarose gel electrophoresis and blotted on CPC-treated nitrocellulose membranes. The polysaccharide composition of these extracts was determined. The membrane stained with toluidine blue (reversible staining) was destained and the same lanes used for immunological detection or other applications. Reversible staining was also applied to recover single species of polysaccharides after electrophoretic separation of mixtures of GAGs and their transfer onto membranes. Single bands were released from the membrane with an efficiency of 70-100% for further biochemical characterization.
DEFF Research Database (Denmark)
Hansen, M.C.; Nielsen, A.K.; Molin, Søren
2001-01-01
obtained by these techniques are compared between experiments in which differences in growth rates, strains, or stress treatments occur, the normalization procedure may have a significant impact on the results. In this report we present a solution to the normalization problem in RNA slot blotting...... the relative level of rRNA per cell, and slot blotting to rRNA probes, which estimates the level of rRNA per extracted total RNA, the amount of RNA per cell was calculated in a series of heat shock experiments with the gram-positive bacterium Lactococcus lactis. It was found that the level of rRNA per cell...... decreased to 30% in the course of the heat shock. This lowered ribosome level led to a decrease in the total RNA content, resulting in a gradually increasing overestimation of the mRNA levels throughout the experiment. Using renormalized cellular mRNA levels, the HrcA-mediated regulation of the genes...
Development of a dot blot assay using gene probes for the detection of enteroviruses in water
International Nuclear Information System (INIS)
Margolin, A.B.
1986-01-01
Enteric viruses are viruses which replicate in the intestinal tract of man and animals. One mode of transmission for enteric viruses is the fecal-oral route. Drinking water which has been contaminated with sewage or sewage effluent has been implicated as a means for the spread of enteric viruses. Current methods for the detection of enteric viruses in water requires the use of animal cell culture. This technique has several drawbacks. More rapid techniques, such as fluorescent antibody or radioimmunoassay do not have the needed sensitivity to detect the low levels of virus found in contaminated water. An alternative technique for the detection of viruses in water was sought. Recent advances in recombinant DNA technology now makes it possible to detect viruses without the use of cell culture or antibodies. Gene probes that hybridize to the RNA of poliovirus and hepatitis A virus were tested for their ability to detect different enteric viruses. The probes were labeled with 32 P dCTP and 32 P dATP to a specific activity greater then 1.0 x 10 9 cpm/ug DNA. One infectious unit of poliovirus and hepatitis A virus was detected using labeled cDNA probes. Upon comparison, the dot blot assay was as sensitive as tissue culture for the detection of poliovirus in beef extract, secondary effluent, and tap water. Environmental samples, such as secondary effluent, reclaimed wastewater and unchlorinated drinking water were also assayed for poliovirus and hepatitis A virus with the use of gene probes. The results presented here offer an alternative method for screening water samples for the presence of enteric viruses
Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
Penna, Aubin; Cahalan, Michael
2007-01-01
Western Blotting (or immunoblotting) is a standard laboratory procedure allowing investigators to verify the expression of a protein, determine the relative amount of the protein present in different samples, and analyze the results of co-immunoprecipitation experiments. In this method, a target protein is detected with a specific primary antibody in a given sample of tissue homogenate or extract. Protein separation according to molecular weight is achieved using denaturing SDS-PAGE. After tr...
Directory of Open Access Journals (Sweden)
Gusti Ngurah Permana
2014-08-01
Full Text Available Pemahaman tentang penentuan jenis kelamin dalam populasi induk merupakan hal yang sangat penting bagi keberhasilan program pembenihan. Pengukuran reaksi antibodi dan aktivitas hormon testosterone, serta estradiol adalah metode dengan potensi yang secara akurat dapat menentukan jenis kelamin ikan tanpa mematikan ikan. Tujuan penelitian ini adalah untuk mengetahui akurasi metode dot blot dan ELISA dengan 11-ketotestorsterone (11-KT yang tersedia secara komersial EIA-kit untuk membedakan jenis kelamin ikan tuna sirip kuning. Hasil analisis menunjukkan bahwa metode dot blot menghasilkan ekspresi vitelogenin tampak jelas pada individu betina dan efek plasma terlihat transparan, jika dibandingkan dengan individu jantan. Interpretasi dari metode ini memerlukan pengalaman dan keahlian dalam akurasi pembacaan hasil. Aktivitas hormon 11-KT dengan sampel klip sirip dan plasma memberikan hasil yang baik dengan aktivitas hormon terlihat jelas.
Lamer, S; Jungblut, P R
2001-03-10
In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.
Kamel, Hanan H; Saad, Ghada A; Sarhan, Rania M
2013-04-01
The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27-(KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.
Proteínas inmunodominantes de Brucella Melitensis evaluadas por Western Blot
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Elizabeth Anaya
1997-01-01
Full Text Available Se separaron extractos de proteínas totales de Brucella melitensis en gel 15% SDS-PAGE. Su seroreactividad fue analizada por Western Blot con resultados satisfactorios. Para éste propósito sueros controles negativos (n=03, sueros de pacientes con brucelosis (n=34, cólera (n=12, tifoidea (n=02 y tuberculosis (n=02 fueron usados. Esta prueba inmunodiagnóstica detectó bandas seroreactivas altamente específicas (100% correspondientes a 8,14,18, un complejo de 25-48 y 58kDa. La sensibilidad del test fue del 90% usando los sueros antes mencionados.
FANCD2 Western blot as a diagnostic tool for Brazilian patients with Fanconi anemia
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D.V. Pilonetto
2009-03-01
Full Text Available Fanconi anemia is a rare hereditary disease showing genetic heterogeneity due to a variety of mutations in genes involved in DNA repair pathways, which may lead to different clinical manifestations. Phenotypic variability makes diagnosis difficult based only on clinical manifestations, therefore laboratory tests are necessary. New advances in molecular pathogenesis of this disease led researchers to develop a diagnostic test based on Western blot for FANCD2. The objective of the present study was to determine the efficacy of this method for the diagnosis of 84 Brazilian patients with Fanconi anemia, all of whom tested positive for the diepoxybutane test, and 98 healthy controls. The FANCD2 monoubiquitinated isoform (FANCDS+/FANCD2L- was not detected in 77 patients (91.7%. In 2 patients (2.4%, there was an absence of both the monoubiquitinated and the non-ubiquitinated proteins (FANCD2S-/FANCD2L- and 5 patients (5.9% had both isoforms (FANCD2S+/FANCD2L+. This last phenotype suggests downstream subtypes or mosaicism. All controls were diepoxybutane negative and were also negative on the FANCD2 Western blot. The Western blot for FANCD2 presented a sensitivity of 94% (79/84 and specificity of 100% (98/98. This method was confirmed as an efficient approach to screen Brazilian patients with deleterious mutations on FANCD2 (FANCD2S-/FANCD2L- or other upstream genes of the FA/BRCA pathway (FANCDS+/FANCD2L-, to confirm the chromosome breakage test and to classify patients according to the level of FA/BRCA pathway defects. However, patients showing both FANCD2 isoforms (FANCD2S+/FANCD2L+ require additional studies to confirm mutations on downstream Fanconi anemia genes or the presence of mosaicism.
International Nuclear Information System (INIS)
Taylor, Ewan R.; Morgan, Iain M.
2003-01-01
Transient DNA replication assays to detect papillomavirus E1/E2-mediated DNA replication have depended upon Southern blotting. This technique is hazardous (radioactive), labour intensive, semiquantitative, and physically limited in the number of samples that can be processed at any one time. We have overcome these problems by developing a real-time PCR protocol for the detection of E1/E2-mediated transient DNA replication. The results demonstrate detection of replication at levels not seen using Southern blotting demonstrating enhanced sensitivity. This technique is also, by definition, highly quantitative. Therefore, the real-time PCR technique is the optimal method for the detection of E1/E2-mediated DNA replication
A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot
DEFF Research Database (Denmark)
Albers, Eliene; Sbroggiò, Mauro; Martin Gonzalez, Javier
2017-01-01
The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific...
Glutathione Transferase GSTπ In Breast Tumors Evaluated By Three Techniques
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Rafael Molina
1993-01-01
Full Text Available The glutathione transferases are involved in intracellular detoxification reactions. One of these, GSTπ, is elevated in some breast cancer cells, particularly cells selected for resistance to anticancer agents. We evaluated GSTπ expression in 60 human breast tumors by three techniques, immunohistochemistry, Northern hybridization, and Western blot analysis. There was a significant positive correlation between the three methods, with complete concordance seen in 64% of the tumors. There was strong, inverse relationship between GSTπ expression and steroid receptor status with all of the techniques utili zed. [n addition, there was a trend toward higher GSTπ expression in poorly differentiated tumors, but no correlation was found between tumor GSTπ content and DNA ploidy or %S-phase. GSTπ expression was also detected in adjacent benign breast tissue as well as infiltrating lymphocytes; this expression may contribute to GSTπ measurements using either Northern hybridization or Western blot analysis. These re sults suggest that immunohistochemistry is the method of choice for measuring GSTπ in breast tumors.
Richards, Mark J; Daniel, Susan
2017-02-07
The supported lipid bilayer has been portrayed as a useful model of the cell membrane compatible with many biophysical tools and techniques that demonstrate its appeal in learning about the basic features of the plasma membrane. However, some of its potential has yet to be realized, particularly in the area of bilayer patterning and phase/composition heterogeneity. In this work, we generate contiguous bilayer patterns as a model system that captures the general features of membrane domains and lipid rafts. Micropatterned polymer templates of two types are investigated for generating patterned bilayer formation: polymer blotting and polymer lift-off stenciling. While these approaches have been used previously to create bilayer arrays by corralling bilayers patches with various types of boundaries impenetrable to bilayer diffusion, unique to the methods presented here, there are no physical barriers to diffusion. In this work, interfaces between contiguous lipid phases define the pattern shapes, with continuity between them allowing transfer of membrane-bound biomolecules between the phases. We examine effectors of membrane domain stability including temperature and cholesterol content to investigate domain dynamics. Contiguous patterning of supported bilayers as a model of lipid rafts expands the application of the SLB to an area with current appeal and brings with it a useful toolset for characterization and analysis. These combined tools should be helpful to researchers investigating lipid raft dynamics and function and biomolecule partitioning studies. Additionally, this patterning technique may be useful for applications such as bioseparations that exploit differences in lipid phase partitioning or creation of membranes that bind species like viruses preferentially at lipid phase boundaries, to name a few.
International Nuclear Information System (INIS)
Lin, P.H.; Selinfreund, R.; Wharton, W.
1986-01-01
Using the western blot technique, they have devised a new procedure that allowed the direct visualization of both the 150KD and the 170KD forms of EGFR by its natural ligand, 125 I-EGF. A431, and placental plasmalemma were purified and solubilized in either SDS-PAGE buffer (without DTT, EDTA) or Triton X-100 (0.5%), resolved on PAGE and electrophoretically transferred onto nitrocellulose (NC) paper. In the absence of boiling, SDS did not denature the EGFR. Although EGER band can be detected after hybridization with 125 I-EGF, the receptor signal was considerably improved with the addition of 0.1% Tween-20. The binding of 125 I-EGF to the both the 150KD and the 170KD bands of the EGFR was specific, reversible and increased with the amount of membrane protein present. The direct visualization of the EGFR using its natural ligand eliminated the necessity for the time consuming antibody preparation. Presently, they are using this technique to identify specific receptors for other ligands
Antibody responses to Borrelia burgdorferi detected by western blot vary geographically in Canada
Ogden, Nicholas H.; Arsenault, Julie; Hatchette, Todd F.; Mechai, Samir; Lindsay, L. Robbin
2017-01-01
Lyme disease is emerging in eastern and central Canada, and most cases are diagnosed using the two-tier serological test (Enzyme Immuno Assay [EIA] followed by Western blot [WB]). Simplification of this algorithm would be advantageous unless it impacts test performance. In this study, accuracy of individual proteins of the IgG WB algorithm in predicting the overall test result in samples from Canadians was assessed. Because Borrelia burgdorferi strains vary geographically in Canada, geographi...
Blotting Assisted by Heating and Solvent Extraction for DESI-MS Imaging
Cabral, Elaine C.; Mirabelli, Mario F.; Perez, Consuelo J.; Ifa, Demian R.
2013-06-01
Imprints of potato sprout ( Solanum tuberosum L.), gingko leaves (Gingko biloba L. ) and strawberries (Fragaria x ananassa Duch. ) were successfully imaged by desorption electrospray ionization mass spectrometry (DESI-MS) on TLC plates through blotting assisted by heating and/or solvent extraction. Ion images showing the distribution of significant compounds such as glycoalkaloid toxins in potato sprout, ginkgolic acids and flavonoids in ginkgo leaves, and sugars and anthocyanidin in strawberry were obtained. Practical implications of this work include analysis of a wide range of irregular or soft materials by different imprinting conditions without requiring the addition of matrices or use of specific kinds of surfaces.
Imaging and high-sensitivity quantification of chemiluminescent labeled DNA-blots
International Nuclear Information System (INIS)
Dorner, G.
1997-01-01
The present thesis has for objective the development of both, methods of DNA labeling by chemiluminescence (via the catalytic activity of the enzyme alkaline phosphatase - AP) and an appropriate imaging system. Offering a competitive alternative to the detection of classical radio-labels in molecular-biological experiments of the blotting type, this technique should permit the realization of quantitative studies of gene expression at ultra-high sensitivity necessary in particular for differential-screening experiments. To reach our aim. we separated the project into three different parts. In a first step an imager based on a liquid-nitrogen-cooled CCD coupled to a standard optics (50 mm/fl.2) has been installed and characterized. This system offers a sensitive area of up to 625 cm 2 , a spatial resolution of 0.3-1 mm (depending on the field of view) and a sensitivity sufficient to detect 10 fg/mm 2 labeled DNA. In a second part, the chemiluminescent light-generation process in solution has been investigated to optimize the parameters temperature. pH and concentration of the substrate as well as the enzyme. The substrate offering the highest light yield (CDP-Star in addition with the enhancer EMERALD II) allows quantification of AP down to 10 -15 M within a dynamic range of 10 4 in solution. Finally. preparation, immobilization and detection of AP-labeled DNA probes (via a biotin-streptavidin-biotin-AP bridge) on nylon membranes has been optimized. A linear relation between the light intensities and the amount of DNA was observed in a range of 10 fg/mm 2 - 100 pg/mm 2 . Hybridization of the probes to bacterial cloned target-DNA has been addressed after examination of the best hybridization conditions. Our protocol includes the treatment of a proteinase, which resulted in a significantly lower background on the filter. The results of our investigations suggest that the main conditions for a reliable differential-screening experiment are fulfilled when using
International Nuclear Information System (INIS)
El-Maghraby, T.K.; Abdelazeim, O.
2002-01-01
The present work has been conducted to determine the mutations related to drug resistance in M. tuberculosis in 63 Egyptian isolates using dot blot hybridization and spoligotyping. The PCR was done for amplification rpoB and katG genes in isolates. Dot blot hybridization were done to PCR products by using specific radiolabelled probes. Moreover, spoligotyping was done to know about the different strains found in Egypt. The results revealed that 58% from isolates had drug resistance to one or more of antituberculosis drugs. The results of spoligotyping have revealed that some Egyptian isolates are identical with the international code while the rest has not been identified yet. DNA sequencing was done to identify the mutation that not clear in dot blot hybridization. Early diagnosis of geno typing resistance to antituberculosis drugs is important as well as allow appropriate early patients management with few days of TB diagnosis. Using such strategy for early diagnosis of TB drug resistance allow and fast and potent patient's management
Nuclear techniques in plant pathology. 2. Use of radioauthography in phytopathology
International Nuclear Information System (INIS)
Silva, D.M.; Lima Nogueira, N. de
1986-01-01
Through association of radioautography techniques and electron or optic microscopy were studied: the sites of virus nucleic acids replication in the tissues plants and protoplasts; interaction bacteriophage-phytopathogenic bacteria; biological functions of Hemileia-coffee plant system. The quantative radioautography allowed distinguish the source of silver grains on two or more organelles and the Southern-Blot technique, with radioative DNA has been used to find the relationship among viruses or plasmids. (Author) [pt
Energy Technology Data Exchange (ETDEWEB)
Malin, E; Belden, E L; Roth, D
1985-09-01
A radioimmunoassay (RIA), an indirect competitive enzyme-linked immunosorbent assay (ELISA), and a dot-blot modification of the ELISA were evaluated for detection and identification of Xanthomonas campestris pv. phaseoli (X. c. pv. phaseoli). RIA and the dot blot tests were specific for X. c. pv. phaseoli; however, significant cross reactions occurred in the indirect competitive ELISA when using anti-X. c. pv. phaseoli antiserum against other closely related bacteria. The sensitivity level of all procedures for X. c. pv. phaseoli was approximately l0/sup 5/ colony forming unitsmL. All procedures were unsatisfactory in reliably detecting low levels of X. c. pv. phaseoli directly from extracts of bean seed. However when used in conjunction with ilution plating the dot blot assay and the RIA would be useful in specifically identifying X. c. pv. phaseoli. The relative merits of these tests for identification of X. c. pv. phaseoli are discussed.
Tsaltas, G; Ford, C H; Gallant, M
1992-01-01
One of the important factors affecting the action of monoclonal antibodies (Mabs) or immunoconjugates on tumour sites depends on whether the Mab is internalized by the cancer cells in question. The underexplored subject of internalization is discussed in this paper, and a number of in vitro techniques for investigating internalization are evaluated, using a model which consists of a well characterized anti-carcinoembryonic antigen (anti-CEA) Mab and a number of CEA expressing human cancer cell lines. Employing two alternative radiolabeling assays, evidence for internalization of the anti-CEA Mab by a CEA-positive colorectal cancer cell line (LS174T) was obtained throughout the time intervals examined (5 min to 150 min). Electronmicroscopy employing horseradish-peroxidase labeled anti-CEA Mab and control antibody permitted direct visualization of anti-CEA Mab-related staining in intracellular compartments of a high CEA-expressor human colorectal cell line (SKCO1). Finally Western blots of samples derived from cytosolic and membrane components of solubilized cells from lung and colonic cancer cell lines provided evidence for internalized anti-CEA Mab throughout seven half hour intervals, starting at 5 minutes. Internalized anti-CEA was detected in all CEA expressing cell lines (LS174T, SKCO1, BENN) but not in the case of a very low CEA expressor line (COLO 320).
He, Dong; Luo, Wen; Wang, Zhiyuan; Lv, Pengmei; Yuan, Zhenhong; Huang, Shaowei; Xv, Jingliang
2017-07-01
Directed evolution has been proved an effective way to improve the stability of proteins, but high throughput screening assays for directed evolution with simultaneous improvement of two or more properties are still rare. In this study, we aimed to establish a membrane-blot assay for use in the high-throughput screening of Rhizomucor miehei lipases (RMLs). With the assistance of the membrane-blot screening assay, a mutant E47K named G10 that showed improved thermal stability was detected in the first round of error-prone PCR. Using G10 as the parent, two variants G10-11 and G10-20 that showed improved thermal stability and methanol tolerance without loss of activity compared to the wild type RML were obtained. The T 50 60 -value of G10-11 and G10-20 increased by 12°C and 6.5°C, respectively. After incubation for 1h, the remaining residual activity of G10-11 and G10-20 was 63.45% and 74.33%, respectively, in 50% methanol, and 15.98% and 30.22%, respectively, in 80% methanol. Thus, we successfully developed a membrane-blot assay that could be used for the high-throughput screening of RMLs with improved thermostability and methanol tolerance. Based on our findings, we believe that our newly developed membrane-blot assay will have potential applications in directed evolution in the future. Copyright © 2017 Elsevier Inc. All rights reserved.
Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization
Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.
1999-01-01
A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324
Flisiak, R; Wierzbicka, I; Prokopowicz, D
1998-01-01
Aim of this study was evaluation of Western blot banding patterns in different clinical forms of early Lyme borreliosis diagnosed in patients from north-eastern Poland, recognized as endemic for tick-borne diseases. Study was performed on serum samples of 48 patients with Lyme borreliosis and 26 healthy volunteers, as controls. Samples tested routinely for total antibody with enzyme immunoassay were subsequently analysed for specific antibodies with Western blot based on antigen extract of European strain of Borrelia burgdorferi. In patients, IgM antibodies were the most frequently directed against 41 kDa and 58 kDa antigens, whereas in control group only antibodies against 45 kDa and 58 kDa were present. Similar response was observed in respect to IgG antibodies. Evaluation of banding pattern in respect to clinical form of the disease revealed the highest prevalence of IgM and IgG anti-41 kDa antibodies in patients with erythema migrans and Lyme arthritis, and anti-58 kDa in neuroborreliosis patients, who had no anti-21 kDa antibodies. Relatively high frequency of IgG antibodies against 21, 30 and 93 kDa antigens was typical for neuroborreliosis. Bands count was significantly higher in different clinical forms of the disease than in controls, and it was the highest in neuroborreliosis. Combined analysis of Western blot results (IgM/IgG) enabled to achieve higher sensitivity (84%) and specificity (100%) than available with the most recommended EIA kits.
Ravera, Ivan; Ferreira, Diana; Gallego, Laia Solano; Bardagí, Mar; Ferrer, Lluís
2015-08-01
The aim of this study was to investigate the presence of canine immunoglobulins (Ig) G against Demodex proteins in the sera of healthy dogs and of dogs with juvenile generalized demodicosis (CanJGD) with or without secondary pyoderma. Demodex mites were collected from dogs with CanJGD. Protein concentration was measured and a western blot technique was performed. Pooled sera from healthy dogs reacted mainly with antigen bands ranging from 55 to 72 kDa. Pooled sera from dogs with CanJGD without secondary pyoderma reacted either with 10 kDa antigen band or 55 to 72 kDa bands. Pooled sera from dogs with CanJGD with secondary pyoderma reacted only with a 10 kDa antigen band. The results of this study suggest that both healthy dogs and dogs with CanJGD develop a humoral response against different proteins of Demodex canis. Copyright © 2015 Elsevier Ltd. All rights reserved.
Energy Technology Data Exchange (ETDEWEB)
Chandler, D.P.; Welt, M.; Leung, F.C.
1990-10-01
An efficient one-step injection technique for gene insertion into fertilized rainbow trout (Oncorhynchus mykiss) eggs is described, and basic parameters affecting egg survival are reported. Freshly fertilized rainbow trout eggs were injected in the perivitelline space with a recombinant mouse metallothionein-genomic bovine growth hormone (bGH) DNA construct using a 30-gauge hypodermic needle and a standard microinjection system. Relative to control, site of injection and DNA concentration did not affect the egg survival, but injections later than 3--4 hours post fertilization were detrimental. The injection technique permitted treatment of 100 eggs/hr with survivals up to 100%, resulting in a 4% DNA uptake rate as indicated by DNA dot blot analysis. Positive dot blot results also indicated that the injected DNA is able to cross the vitelline membrane and persist for 50--60 days post hatching, obviating the need for direct injection into the germinal disk. Results are consistent with previous transgenic fish work, underscoring the usefulness of the technique for generating transgenic trout and salmonids. 24 refs., 6 figs., 3 tabs.
Liance, Martine; Janin, Veronique; Bresson-Hadni, Solange; Vuitton, Dominique-Angele; Houin, Rene; Piarroux, Renaud
2000-01-01
The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis-infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species-specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID:11015390
Should we ignore western blots when selecting antibodies for other applications?
DEFF Research Database (Denmark)
Uhlén, Mathias
2017-01-01
.In the Human Protein Atlas (HPA) program, we have validated more than 24,000 in-house-generated antibodies directed to 17,000 human target proteins2. Although there is often a correlation between performance in different applications, we have observed many examples of antibodies that show strong support...... applications and that this influences the epitopes exposed on the target protein, which might have profound consequences for the ability of a given antibody to bind specifically to its target. As an example, proteins that are analyzed by immunohistochemistry (IHC) are normally first cross-linked with formalin.......In conclusion, western blot and protein array analyses can indeed be useful tools when selecting specific antibodies for other applications. The use of these methods is encouraged both for antibody providers and users, and antibodies with signs of cross-reactivity in these applications should be treated...
Mere end blot en bid af hverdagen- Måltidet i et leve- og bomiljø
DEFF Research Database (Denmark)
Bundgaard, Karen Marie
2005-01-01
. Datamaterialet bygger på deltagerobservationer og interviews. Undersøgelsen viste, at den måde måltiderne var organiseret på gav tid og rum til en hjemlig atmosfære, til et levende fællesskab, til det at være noget og at være sig selv og til at have værdifulde gøremål. Måltiderne var ikke blot en bid – men en...
Pal, Jayanta K; Berwal, Sunil K; Soni, Rupali N
2012-01-01
A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS) polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water and viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining), and cost-effective.
Millar, Thomas James; Knighton, Ronald; Chuck, Jo-Anne
2012-01-01
Immunological detection of proteins is an essential method to demonstrate to undergraduate biology students, however, is often difficult in resource and time poor student laboratory sessions. This method describes a failsafe method to rapidly and economically demonstrate this technique using biotinylated proteins or biotin itself as targets for…
Maagd, de R.A.; Klei, van der H.; Bakker, P.L.; Stiekema, W.J.; Bosch, D.
1996-01-01
We investigated the role of the constituent domains of the CryIA(b) and CryIA(c) δ-endotoxins in binding to midgut epithelial cell membrane proteins of Spodoptera exigua and Manduca sexta on ligand blots. A collection of wild- type and CryIC-CryIA hybrid toxins was used for this purpose. As
Directory of Open Access Journals (Sweden)
Eduardo Miranda-Ulloa
Full Text Available Con el objetivo de evaluar las pruebas dot blot y aglutinación de látex para la detección de cisticercosis humana con antígeno de líquido de cisticerco de Taenia solium, se usaron 125 sueros humanos, de los cuales 60 procedían de personas con cisticercosis confirmada por Western Blot, 45 de personas con otras enfermedades parasitarias y 20 de personas aparentemente sanas. La concentración óptima del antígeno para impregnar las tiras dot blot fue de 0,01 ug/uL, y para impregnar las partículas de látex fue de 0,092 ug/uL. Para la prueba dot blot se encontró una sensibilidad del 100% y especificidad del 87,7%; para la aglutinación de látex una sensibilidad del 93,3% y especificidad del 89,2%. Ambas pruebas podrían ser de utilidad y factibles de implementar como alternativas de diagnóstico serológico en laboratorios de áreas endémicas del Perú
Hood-Degrenier, Jennifer K
2008-01-01
The movement of newly synthesized proteins through the endomembrane system of eukaryotic cells, often referred to generally as the secretory pathway, is a topic covered in most intermediate-level undergraduate cell biology courses. An article previously published in this journal described a laboratory exercise in which yeast mutants defective in two distinct steps of protein secretion were differentiated using a genetic reporter designed specifically to identify defects in the first step of the pathway, the insertion of proteins into the endoplasmic reticulum (Vallen, 2002). We have developed two versions of a Western blotting assay that serves as a second way of distinguishing the two secretory mutants, which we pair with the genetic assay in a 3-wk laboratory module. A quiz administered before and after students participated in the lab activities revealed significant postlab gains in their understanding of the secretory pathway and experimental techniques used to study it. A second survey administered at the end of the lab module assessed student perceptions of the efficacy of the lab activities; the results of this survey indicated that the experiments were successful in meeting a set of educational goals defined by the instructor.
Directory of Open Access Journals (Sweden)
Kun Ismiyatin
2010-06-01
Full Text Available Background: There are several dentin surface drying techniques to provide a perfect resin penetration on dentin. There are two techniques which will be compared in this study. The first technique was by rubbing dentin surface gently using cotton pellet twice, this technique is called blot dry technique. The second technique is by air blowing dentin surface for one second and continued by rubbing dentin surface gently using moist cotton. Purpose: This experiment was aimed to examine the best dentin surface drying techniques after 37% phosphoric acid etching to obtain the optimum tensile bond strength between hydroxyethyl methacrylate (HEMA and dentin surface. Method: Bovine teeth was prepared flat to obtain the dentin surface and than was etched using 37% phosphoric acid for 15 seconds. After etching the dentin was cleaned using 20 cc plain water and dried with blot dry techniques (group I, or dried with air blow for one second (group II, or dried with air blow for one second, and continued with rubbing gently using moist cotton pellet (group III, and without any drying as control group (group IV. After these drying, the dentin surfaces were applied with resin dentin bonding agent and put into plunger facing the composite mould. The antagonist plunger was filled with composite resin. After 24 hours, therefore bond strength was measured using Autograph. Result: Data obtained was analyzed using One-Way ANOVA with 95% confidence level and continued with LSD test on p≤0.05. The result showed that the highest tensile bond strength was on group I, while the lowest on group IV. Group II and IV, III and IV, II and III did not show signigicant difference (p>0.05. Conclusion: Dentin surface drying techniques through gentle rubbing using cotton pellet twice (blot dry technique gave the greatest tensile bond strength.Latar belakang masalah: Tehnik pengeringan permukaan dentin agar resin dapat penetrasi dengan sempurna adalah dengan cara pengusapan secara
Rutgers, Abraham; Damoiseaux, Jan; Roozendaal, Caroline; Limburg, Pieter C; Stegeman, Coen A; Tervaert, Jan Willem Cohen
Rapidly progressive glomerulonephritis (RPGN) is characterized by rapid and progressive loss of renal function and the presence of crescentic glomerulonephritis (CGN). Early diagnosis and appropriate treatment is mandatory to prevent death and/or renal failure. We have evaluated an ANCA-GBM dot-blot
Directory of Open Access Journals (Sweden)
Nilton Thaumaturgo
2002-10-01
Full Text Available Sm14 was the first fatty acid-binding protein homologue identified in helminths. Thereafter, members of the same family were identified in several helminth species, with high aminoacid sequence homology between them. In addition, immune crossprotection was also reported against Fasciola hepatica infection, in animals previously immunized with the Schistosoma mansoni vaccine candidate, r-Sm14. In the present study, data on preliminary sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis of nine different helminth extracts focusing the identification of Sm14 related proteins, is reported. Out of these, three extracts - Ascaris suum (males and females, Echinostoma paraensei, and Taenia saginata - presented components that comigrated with Sm14 in SDS-PAGE, and that were recognized by anti-rSm14 policlonal serum, in Western blotting tests.
Directory of Open Access Journals (Sweden)
Maite Sabalza
Full Text Available In recent years, there have been increasing numbers of infectious disease outbreaks that spread rapidly to population centers resulting from global travel, population vulnerabilities, environmental factors, and ecological disasters such as floods and earthquakes. Some examples of the recent outbreaks are the Ebola epidemic in West Africa, Middle East respiratory syndrome coronavirus (MERS-Co in the Middle East, and the Zika outbreak through the Americas. We have created a generic protocol for detection of pathogen RNA and/or DNA using loop-mediated isothermal amplification (LAMP and reverse dot-blot for detection (RDB and processed automatically in a microfluidic device. In particular, we describe how a microfluidic assay to detect HIV viral RNA was converted to detect Zika virus (ZIKV RNA. We first optimized the RT-LAMP assay to detect ZIKV RNA using a benchtop isothermal amplification device. Then we implemented the assay in a microfluidic device that will allow analyzing 24 samples simultaneously and automatically from sample introduction to detection by RDB technique. Preliminary data using saliva samples spiked with ZIKV showed that our diagnostic system detects ZIKV RNA in saliva. These results will be validated in further experiments with well-characterized ZIKV human specimens of saliva. The described strategy and methodology to convert the HIV diagnostic assay and platform to a ZIKV RNA detection assay provides a model that can be readily utilized for detection of the next emerging or re-emerging infectious disease.
Itai, Shunsuke; Kaneko, Mika K; Fujii, Yuki; Yamada, Shinji; Nakamura, Takuro; Yanaka, Miyuki; Saidoh, Noriko; Handa, Saori; Chang, Yao-Wen; Suzuki, Hiroyoshi; Harada, Hiroyuki; Kato, Yukinari
2017-10-01
The epidermal growth factor receptor (EGFR) is a member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases and is involved in cell growth and differentiation. EGFR homodimers or heterodimers with other HER members, such as HER2 and HER3, activate downstream signaling cascades in many cancers. In this study, we developed novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. First, we expressed the full-length or ectodomain of EGFR in LN229 glioblastoma cells and then immunized mice with LN229/EGFR or ectodomain of EGFR, and performed the first screening using enzyme-linked immunosorbent assays. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical (fourth screening) analyses. Among 100 mAbs, only one clone EMab-51 (IgG 1 , kappa) reacted with EGFR in Western blot analysis. Finally, immunohistochemical analyses with EMab-51 showed sensitive and specific reactions against oral cancer cells, warranting the use of EMab-51 to detect EGFR in pathological analyses of EGFR-expressing cancers.
Cy5 total protein normalization in Western blot analysis.
Hagner-McWhirter, Åsa; Laurin, Ylva; Larsson, Anita; Bjerneld, Erik J; Rönn, Ola
2015-10-01
Western blotting is a widely used method for analyzing specific target proteins in complex protein samples. Housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. We present a new, more reliable method for normalization using Cy5-prelabeled total protein as a loading control. We used a prelabeling protocol based on Cy5 N-hydroxysuccinimide ester labeling that produces a linear signal response. We obtained a low coefficient of variation (CV) of 7% between the ratio of extracellular signal-regulated kinase (ERK1/2) target to Cy5 total protein control signals over the whole loading range from 2.5 to 20.0μg of Chinese hamster ovary cell lysate protein. Corresponding experiments using actin or tubulin as controls for normalization resulted in CVs of 13 and 18%, respectively. Glyceraldehyde-3-phosphate dehydrogenase did not produce a proportional signal and was not suitable for normalization in these cells. A comparison of ERK1/2 signals from labeled and unlabeled samples showed that Cy5 prelabeling did not affect antibody binding. By using total protein normalization we analyzed PP2A and Smad2/3 levels with high confidence. Copyright © 2015 Elsevier Inc. All rights reserved.
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Aravalli, Rajagopal N., E-mail: aravalli@umn.edu [Department of Radiology, University of Minnesota Medical School, MMC 292, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Park, Chang W. [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Steer, Clifford J., E-mail: steer001@umn.edu [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455 (United States)
2016-08-26
The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type.
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Aravalli, Rajagopal N.; Park, Chang W.; Steer, Clifford J.
2016-01-01
The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type.
TSE strain differentiation in mice by immunohistochemical PrP(Sc) profiles and triplex Western blot.
van Keulen, Lucien J M; Langeveld, Jan P M; Dolstra, Corry H; Jacobs, Jorg; Bossers, Alex; van Zijderveld, Fred G
2015-10-01
TSE strains are routinely identified by their incubation period and vacuolation profile in the brain after intracerebral inoculation and serial passaging in inbred mouse lines. There are some major drawbacks to this method that are related to the variation in vacuolation that exists in the brains of mice infected with the same TSE strain and to variation between observers and laboratories in scoring vacuolation and determining the final incubation period. We investigated the potential of PrP(Sc) immunohistochemistry and triplex Western blotting as possible alternative methods to differentiate between TSE strains. TSE reference strains ME7, 87A/87V, 22A/22C, 79A/79V and 301C/301V were intracerebrally inoculated in RIII or VM inbred mice that differ in their PrP genotype. Immunohistochemical PrP(Sc) profiles were drawn up by scanning light microscopy both on coronal and sagittal sections. On the basis of the localization of PrP(Sc) in the cerebral cortex, hippocampus, and cerebellar cortex and the overall type of PrP(Sc) staining, all TSE strains could be well differentiated from each other through their typical strain dependent characteristics. In addition, Western blot showed that the combination of glycosylation profile and 12B2 epitope content of PrP(Sc) allowed to distinguish between all reference strains except for ME7 and 22A in VM mice. TSE strains in mice can be identified on the basis of their PrP(Sc) profile alone. The potential to identify TSE strains in ruminants with these PrP(Sc) profiles after a single primary passage in mice will be the topic of future studies. © 2014 British Neuropathological Society.
Patel, Vyomesh; Ramesh, Arun; Traicoff, June L; Baibakov, Galina; Emmert-Buck, Michael R; Gutkind, J Silvio; Knezevic, Vladimir
2005-05-01
Given the role of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinomas (HNSCC), several rational approaches have now been utilized to abrogate tyrosine kinase activity and its disengagement from downstream signal transducers. Monitoring the activity of these molecules could potentially be useful to determine not only drug efficacy but also to identify HNSCC patients most likely to benefit from this type of therapy. In this study we have used a novel high throughput multi-layered Western blotting (MLWestern) method that allows the detection of multiple proteins from a single experiment in order to characterize key components in the EGFR signaling pathway in HNSCC cells. Total and activated forms of EGFR and the downstream effectors, Erk and Akt were readily detected in HNSCC cells, where in the control cells (HaCaT) these proteins could only be detected in EGF stimulated cells. Results from conventional Western blot and MLWestern were comparable. Clustering analysis of protein expression revealed similarities in cellular response between some of the cell lines indicative of similarities in their biological response. The data indicate that MLWestern can be potentially applied to identify molecular targets that could be used for rational therapeutic intervention strategies.
Xiao, Xiao; Wu, Honghong; Zhou, Xinghu; Xu, Sheng; He, Jian; Shen, Wenbiao; Zhou, Guanghong; Huang, Ming
2012-06-01
With the widespread use of Roundup Ready soy (event 40-3-2) (RRS), the comprehensive detection of genetically modified component in foodstuffs is of significant interest, but few protein-based approaches have been found useful in processed foods. In this report, the combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different RRS plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing both meat and plant protein concentrates. The validity of the 2 methods was confirmed first. We also showed that the CP4-EPSPS protein existed in different RRS plant tissues. In certain cases, the results from the western blot and the qPCR were not consistent. To be specific, at least 2 degraded fragments of CP4-EPSPS protein (35.5 and 24.6 kDa) were observed. For dried bean curd crust and deep-fried bean curd, a degraded protein fragment with the size of 24.6 kDa appeared, while cp4-epsps gene could not be traced by qPCR. In contrast, we found a signal of cp4-epsps DNA in 3 foodstuffs, including soy-containing ham cutlet product, meat ball, and sausage by qPCR, while CP4-EPSPS protein could not be detected by western blot in such samples. Our study therefore concluded that the combination of DNA- and protein-based methods would compensate each other, thus resulting in a more comprehensive detection from nucleic acid and protein levels. The combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different Roundup Ready soy (event 40-3-2) plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing a combination of both meat and plant protein concentrates. This study indicated that the combination of DNA- and protein-based methods
Martel, Clothilde; Vignaud, Guillaume; Liozon, Eric; Magy, Laurent; Gallouedec, Gael; Ly, Kim; Bezanahary, Holly; Cypierre, Anne; Lapébie, François-Xavier; Palat, Sylvain; Gondran, Guillaume; Jauberteau, Marie-Odile; Fauchais, Anne-Laure
2016-01-01
Idiopathic inflammatory myopathies (IIM) are heterogeneous autoimmune diseases with wide clinical spectrum that may lead to delayed diagnosis. The aim of this study was to examine the impact of IIM-specific dot-blot assay on diagnostic process of patients presenting with muscular or systemic symptoms evocating of IIM. We collected all the prescriptions of an IIM specific dot-blot assay (8 autoantigens including Jo-1, PL-7, PL-12, SRP, Mi-2, Ku, PM/Scl and Scl-70) over a 38-month period. 316 myositis dot-blot assays (MSD) were performed in 274 patients (156 women, mean age 53±10.6 years) referring for muscular and/or systemic symptoms suggesting IIM. The timing of dot prescription through the diagnostic process was highly variable: without (35%), concomitantly (16%) or after electromyographic studies (35%). Fifty-nine patients (22%) had IIM according to Bohan and Peter's criteria. Among them, 29 (49%) had positive dot (8 Jo-1, 6 PM-Scl, 5 PL-12, 5 SRP, 2 Mi-2, 2 PL-7 and 1 Ku). Various other diagnoses were performed including 35 autoimmune disease or granulomatosis (12%), 19 inflammatory rheumatic disease (7%), 16 non inflammatory muscular disorders (6%), 10 drug-induced myalgia (4%), 11 infectious myositis (4%). Except 11 borderline SRP results and one transient PM-Scl, MSD was positive only in one case of IIM. Dot allowed clinicians to correct diagnosis in 4 cases and improved the diagnosis of IIM subtypes in 4 cases. This study reflects the interest of myositis dot in the rapid diagnosis process of patients with non-specific muscular symptoms leading to various diagnoses including IIM.
Characterization of Nora Virus Structural Proteins via Western Blot Analysis.
Ericson, Brad L; Carlson, Darby J; Carlson, Kimberly A
2016-01-01
Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses.
Rosette Assay: Highly Customizable Dot-Blot for SH2 Domain Screening.
Ng, Khong Y; Machida, Kazuya
2017-01-01
With a growing number of high-throughput studies, structural analyses, and availability of protein-protein interaction databases, it is now possible to apply web-based prediction tools to SH2 domain-interactions. However, in silico prediction is not always reliable and requires experimental validation. Rosette assay is a dot blot-based reverse-phase assay developed for the assessment of binding between SH2 domains and their ligands. It is conveniently customizable, allowing for low- to high-throughput analysis of interactions between various numbers of SH2 domains and their ligands, e.g., short peptides, purified proteins, and cell lysates. The binding assay is performed in a 96-well plate (MBA or MWA apparatus) in which a sample spotted membrane is incubated with up to 96 labeled SH2 domains. Bound domains are detected and quantified using a chemiluminescence or near-infrared fluorescence (IR) imaging system. In this chapter, we describe a practical protocol for rosette assay to assess interactions between synthesized tyrosine phosphorylated peptides and a library of GST-tagged SH2 domains. Since the methodology is not confined to assessment of SH2-pTyr interactions, rosette assay can be broadly utilized for ligand and drug screening using different protein interaction domains or antibodies.
Coyne, Cody P; Rashmir-Raven, Ann; Jones, Toni; Mochal, Cathleen; Linford, Robert L; Brashier, Michael; Eddy, Alison
2009-01-01
Limited research to date has characterized the potential for HRPO to function as a primary molecular probe. Pulmonary airway fluid was developed by non-reducing far-Western (ligand) blot analyses utilizing conjugated HRPO-strepavidin or non-conjugated HRPO without the presence of primary immunoglobulin. Endogenous esterase-like biochemical activity of fractions within pulmonary airway fluid was inactivated to determine if they were capable of biochemically converting HRPO chemiluminescent substrate. Complementary analyses modified pulmonary fluid and HRPO with beta-galactosidase and alpha-mannosidase respectively, in addition to determining the influence of mannose and maltose competitive binding on HRPO far-Western (ligand) blot analyses. Identification of pulmonary fluid fractions detected by HRPO far-Western blot analyses was determined by mass spectrometry. Modification of pulmonary fluid with beta-galactosidase, and HRPO with alpha-mannosidase in concert with maltose and mannose competitive binding analyses altered the intensity and spectrum of pulmonary fluid fractions detected by HRPO far-Western blot analysis. Identity of pulmonary airway fluid fractions detected by HRPO far-Western (ligand) blot analysis were transferrin, dynein, albumin precursor, and two 156 kDa equine peptide fragments. HRPO can function as a partially-selective primary molecular probe when applied in either a conjugated or non-conjugated form. Some protein fractions can form complexes with HRPO through molecular mechanisms that involve physical interactions at the terminal alpha-mannose-rich regions of HRPO glycan side-chains. Based on its known molecular composition and structure, HRPO provides an opportunity for the development of diagnostics methodologies relevant to disease biomarkers that possess mannose-binding avidity.
Comparison of Multispot EIA with Western blot for confirmatory serodiagnosis of HIV.
Torian, Lucia V; Forgione, Lisa A; Punsalang, Amado E; Pirillo, Robert E; Oleszko, William R
2011-12-01
Recent improvements in the sensitivity of immunoassays (IA) used for HIV screening, coupled with increasing recognition of the importance of rapid point-of-care testing, have led to proposals to adjust the algorithm for serodiagnosis of HIV so that screening and confirmation can be performed using a dual or triple IA sequence that does not require Western blotting for confirmation. One IA that has been proposed as a second or confirmatory test is the Bio-Rad Multispot(®) Rapid HIV-1/HIV-2 Test. This test would have the added advantage of differentiating between HIV-1 and HIV-2 antibodies. To compare the sensitivity and type-specificity of an algorithm combining a 3rd generation enzyme immunoassay (EIA) followed by a confirmatory Multispot with the conventional algorithm that combines a 3rd generation EIA (Bio-Rad GS HIV-1/HIV-2 Plus O EIA) followed by confirmatory Western blot (Bio-Rad GS HIV-1 WB). 8760 serum specimens submitted for HIV testing to the New York City Public Health Laboratory between May 22, 2007, and April 30, 2010, tested repeatedly positive on 3rd generation HIV-1-2+O EIA screening and received parallel confirmatory testing by WB and Multispot (MS). 8678/8760 (99.1%) specimens tested WB-positive; 82 (0.9%) tested WB-negative or indeterminate (IND). 8690/8760 specimens (99.2%) tested MS-positive, of which 14 (17.1%) had been classified as negative or IND by WB. Among the HIV-1 WB-positive specimens, MS classified 26 (0.29%) as HIV-2. Among the HIV-1 WB negative and IND, MS detected 12 HIV-2. MS detected an additional 14 HIV-1 infections among WB negative or IND specimens, differentiated 26 HIV-1 WB positives as HIV-2, and detected 12 additional HIV-2 infections among WB negative/IND. A dual 3rd generation EIA algorithm incorporating MS had equivalent HIV-1 sensitivity to the 3rd generation EIA-WB algorithm and had the added advantage of detecting 12 HIV-2 specimens that were not HIV-1 WB cross-reactors. In this series an algorithm using EIA
Immunological diagnosis of human hydatid cyst using Western immunoblotting technique
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Mahboubeh Hadipour
2016-01-01
Full Text Available Background: Echinococcosis is a parasitic disease with worldwide distribution which is caused by the tapeworms Echinococcus granulosus. Diagnosis of the disease relies on imaging techniques, but the techniques are not able to differentiate the cyst from benign or malignant tumors; hence, appropriate serologic methods are required for the differential diagnosis of the infection. Materials and Methods: In this investigation, different sheep hydatid cyst antigens probed with thirty sera of patients with hydatid cyst and also thirty human normal sera using Western immunoblotting technique. Considering results of surgery as gold standard, sensitivity and specificity of Western blotting was estimated. Results: Sera of 29, 26, and 16 patients with hydatid cyst reacted with specific bands of hydatid cyst fluid (HCF, protoscolex crude antigen, and cyst wall crude antigen, respectively. However, none of the normal human sera reacted with those specific bands. Conclusion: A 20 kDa band of sheep HCF is an appropriate antigen for serodiagnosis of hydatid cyst infection.
Sánchez, B; Monteón, V; Reyes, P A; Espinoza, B
2001-01-01
This report describes two assays for the detection of anti-Trypanosoma cruzi antibodies using Mexican strains of the parasite and the concordance with two assays previously evaluated at the Instituto Nacional de Cardiología Ignacio Chávez in Mexico City. Micro-enzyme-linked immunosorbent assay (ELISA) and Western blot were used for the detection of T. cruzi antibodies with a total extract of epimastigote from Ninoa and Queretaro, which are Mexican strains of T. cruzi. To standardize these methods, a total of 246 serum samples was used. In addition, sera from six confirmed Mexican chronic individuals in the asymptomatic phase were also used for comparison with the Argentinean antigen. ELISA was 100% specific in that no false positive results were found with sera of both healthy individuals and non-Chagasic cardiopaths. Sera from individuals infected with Leishmania sp. showed approximately 16% of cross-reaction with ELISA. The test showed a positive predictive value of 90% and a negative predictive value of 100%. Western blot was also a highly sensitive test for detecting chronic Chagasic symptomatic patients from Mexico because no false negative results were obtained. Furthermore, it was possible to use Western blot to detect seven immunodominant antigens of approximately 30, 32, 40, 42, 65, 70, and 83 kDa. Concordance with two previous standardized tests at the Instituto Nacional de Cardiología showed a Kappa index of 0.96, indicating high concordance between the results obtained at these two laboratories. Finally, ELISA using Ninoa antigen extract was more sensitive than ELISA with an Argentinean extract, which failed to detect individuals in the chronic asymptomatic phase (undetermined phase) of infection. This study indicates that ELISA and Western blot using Ninoa and/or Queretaro extracts of T. cruzi as antigens are useful tools in the detection of individuals who have been exposed to T. cruzi both in the undetermined/asymptomatic and symptomatic phases
Li, Jingyi; Liu, Qian; Alsamarri, Hussein; Lounsbury, Jenny A; Haversitick, Doris M; Landers, James P
2013-03-07
Reliable measurement of DNA concentration is essential for a broad range of applications in biology and molecular biology, and for many of these, quantifying the nucleic acid content is inextricably linked to obtaining optimal results. In its most simplistic form, quantitative analysis of nucleic acids can be accomplished by UV-Vis absorbance and, in more sophisticated format, by fluorimetry. A recently reported new concept, the 'pinwheel assay', involves a label-free approach for quantifying DNA through aggregation of paramagnetic beads in a rotating magnetic field. Here, we describe a simplified version of that assay adapted for execution using only a pipet and filter paper. The 'pipette, aggregate, and blot' (PAB) approach allows DNA to induce bead aggregation in a pipette tip through exposure to a magnetic field, followed by dispensing (blotting) onto filter paper. The filter paper immortalises the extent of aggregation, and digital images of the immortalized bead conformation, acquired with either a document scanner or a cell phone camera, allows for DNA quantification using a noncomplex algorithm. Human genomic DNA samples extracted from blood are quantified with the PAB approach and the results utilized to define the volume of sample used in a PCR reaction that is sensitive to input mass of template DNA. Integrating the PAB assay with paper-based DNA extraction and detection modalities has the potential to yield 'DNA quant-on-paper' devices that may be useful for point-of-care testing.
Trenfield, K; Salmond, C A; Pope, J H; Hardie, I R
1993-01-01
The 104 skin biopsies from 34 patients who attended a Renal Transplant Unit in Brisbane over 12 months included 40 squamous cell carcinoma (SCC), 22 solar keratoses, 4 hyperkeratoses, 18 warts and 11 basal cell carcinoma (BCC). Human papillomavirus (HPV) DNA was identified by Southern blot hybridisation using, as individual probes, purified insert DNA from recombinant HPV 1, 2, 3 or 3/10, 4, 5 or 5/8, 7, 11, 16, 18 and 41 under relaxed conditions and characterised by restriction enzyme analysis and Southern blot hybridisation under more stringent conditions. Genomic HPV DNA was characterised in 7 skin biopsies from 4 renal allograft recipients (RARs): HPV 1A in a SCC (20 copies/cell) and a BCC (10 copies/cell) from the one patient, HPV 36 (20 copies/cell) in a SCC, HPV 1A [symbol: see text] 1000 copies/cell) in a wart and HPV 2B (200-800 copies/cell) in 3 warts from the one patient. Only HPV 1A in the SCC exhibited a significant degree of subtype variation. HPV DNA was identified in another 5 skin biopsies from another 4 RARs: HPV 3A in a wart and a hyperkeratosis, HPV 3/10-related DNA in 2 solar keratoses and HPV 5/8-related DNA in another (20-50 copies/cell). The incidence of HPV 5 (or 5-related HPVs) in RAR SCC was very low and that of HPV DNA in RAR warts was lower than that recorded elsewhere but this was not due to insensitivity of the assays. There was no evidence for a role for HPV in the aetiology of skin cancer in RARs in south-eastern Queensland but the possibility remains that as yet unidentified HPV types are involved.
Detection of anti-HIV-1 IgG antibodies in whole saliva by GACELISA and Western blot assays.
Matee, M I; Lyamuya, E F; Simon, E; Mbena, E C; Kagoma, C; Samaranayake, L P; Scheutz, F
1996-05-01
The present study, based on 158 HIV seropositives and 167 HIV seronegatives, demonstrates that saliva collected with the Omni-SAL device and tested with GACELISA (an IgG antibody capture ELISA) is an effective non-invasive alternative to serum for anti-HIV IgG antibody screening. The study also shows that a conventional serum Western blot kit can be used, with slight modifications, for confirmatory testing of saliva specimens. Collecting saliva with the Omni-SAL device had a very good acceptance rate among Tanzanian subjects, and although this diagnostic method is not yet known by the general public, 65% of the study participants preferred to give saliva instead of blood for HIV testing.
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Garth L. Nicolson
2012-03-01
Full Text Available Background: An open label 8-week preliminary study was conducted in a small number of patients to determine if a combination oral supplement containing a mixture of phosphoglycolipids, coenzyme Q10 and microencapsulated NADH and other nutrients could affect fatigue levels in long-term, Western blot-positive, multi-symptom ‘chronic Lyme disease’ patients (also called ‘post-treatment Lyme disease’ or ‘post Lyme syndrome’ with intractable fatigue. Methods: The subjects in this study were 6 males (mean age = 45.1 ± 12.4 years and 10 females (mean age = 54.6 ± 7.4 years with ‘chronic Lyme disease’ (determined by multiple symptoms and positive Western blot analysis that had been symptomatic with chronic fatigue for an average of 12.7 ± 6.6 years. They had been seen by multiple physicians (13.3 ± 7.6 and had used many other remedies, supplements and drugs (14.4 ± 7.4 without fatigue relief. Fatigue was monitored at 0, 7, 30 and 60 days using a validated instrument, the Piper Fatigue Scale.Results: Patients in this preliminary study responded to the combination test supplement, showing a 26% reduction in overall fatigue by the end of the 8-week trial (p< 0.0003. Analysis of subcategories of fatigue indicated that there were significant improvements in the ability to complete tasks and activities as well as significant improvements in mood and cognitive abilities. Regression analysis of the data indicated that reductions in fatigue were consistent and occurred with a high degree of confidence (R2= 0.998. Functional Foods in Health and Disease 2012, 2(3:35-47 Conclusions: The combination supplement was a safe and effective method to significantly reduce intractable fatigue in long-term patients with Western blot-positive ‘chronic Lyme disease.’
Hsiao, K M; Lin, H M; Pan, H; Li, T C; Chen, S S; Jou, S B; Chiu, Y L; Wu, M F; Lin, C C; Li, S Y
1999-01-01
Myotonic dystrophy (DM) is caused by a CTG trinucleotide expansion mutation at exon 15 of the myotonic dystrophy protein kinase gene. The clinical severity of this disease correlates with the length of the CTG trinucleotide repeats. Determination of the CTG repeat length has been primarily relied on by Southern blot analysis of restriction enzyme-digested genomic DNA. The development of PCR-based Southern blotting methodology provides a much more sensitive and simpler protocol for DM diagnosis. However, the quality of the template and the high (G+C) ratio of the amplified region hamper the use of PCR on the diagnosis of DM. A modified PCR protocol to amplify different lengths of CTG repeat region using various concentrations of 7deaza-dGTP has been reported (1). Here we describe a procedure including sample collection, DNA purification, and PCR analysis of CTG repeat length without using 7-deaza-dGTP. This protocol is very sensitive and convenient because only a small number of nucleate cells are needed for detection of CTG expansion. Therefore, it could be very useful in clinical and prenatal diagnosis as well as in prevalence study of DM.
International Nuclear Information System (INIS)
SenGupta, D.N.; Kumar, P.; Zmudzka, B.Z.; Coughlin, S.; Vishwanatha, J.K.; Robey, F.A.; Parrott, C.; Wilson, S.H.
1987-01-01
A new polyclonal antibody against the α-polymerase catalytic polypeptide was prepared by using homogeneous HeLa cellα-polymerase. The antibody neutralized α-polymerase activity and was strong and specific for the α-polymerase catalytic polypeptide (M/sub r/ 183,000) in Western blot analysis of crude extracts of HeLa cells. The antibody was used to screen a cDNA library of newborn rat brain poly(A+) RNA in λgt11. A positive phage was identified and plaque purified. This phage, designated λpolα1.2, also was found to be positive with an antibody against Drosophila α-polymerase. The insert in λpolα1.2 (1183 base pairs) contained a poly(A) sequence at the 3' terminus and a short in-phase open reading frame at the 5' terminus. A synthetic oligopeptide (eight amino acids) corresponding to the open reading frame was used to raise antiserum in rabbits. Antibody affinity purified from this serum was found to be immunoreactive against purified α-polymerase by enzyme-linked immunosorbent assay and was capable of immunoprecipitating α-polymerase. This indicated the λpolα1.2 insert encoded an α-polymerase epitope and suggested that the cDNA corresponded to an α-polymerase mRNA. This was confirmed in hybrid selection experiments using pUC9 containing the cDNA insert and poly(A+) RNA from newborn rat brain; the insert hybridized to mRNA capable of encoding α-polymerase catalytic polypeptides. Northern blot analysis of rat brain poly(A+) RNA revealed that this mRNA is ∼5.4 kilobases
Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research
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Helena Kupcova Skalnikova
2017-12-01
Full Text Available Melanoma is a skin cancer with permanently increasing incidence and resistance to therapies in advanced stages. Reports of spontaneous regression and tumour infiltration with T-lymphocytes makes melanoma candidate for immunotherapies. Cytokines are key factors regulating immune response and intercellular communication in tumour microenvironment. Cytokines may be used in therapy of melanoma to modulate immune response. Cytokines also possess diagnostic and prognostic potential and cytokine production may reflect effects of immunotherapies. The purpose of this review is to give an overview of recent advances in proteomic techniques for the detection and quantification of cytokines in melanoma research. Approaches covered span from mass spectrometry to immunoassays for single molecule detection (ELISA, western blot, multiplex assays (chemiluminescent, bead-based (Luminex and planar antibody arrays, ultrasensitive techniques (Singulex, Simoa, immuno-PCR, proximity ligation/extension assay, immunomagnetic reduction assay, to analyses of single cells producing cytokines (ELISpot, flow cytometry, mass cytometry and emerging techniques for single cell secretomics. Although this review is focused mainly on cancer and particularly melanoma, the discussed techniques are in general applicable to broad research field of biology and medicine, including stem cells, development, aging, immunology and intercellular communication.
Huguet, S; Sghiri, R; Ballot, E; Johanet, C
2004-01-01
The Cyto-Dot 4 HM043 kit commercialised by BMD, has replaced the Cyto-Dot HM010 kit that allowed three auto-antibodies detection (anti-Jo-1, anti-M2 and anti-ribosomal protein). Detection of anti-LKM1 auto-antibody was added. These four auto-antibodies have in common only the intracytoplasmic localisation of their respective antigen. The aim of our study was to evaluate this new kit using 104 sera and to compare our results with reference techniques (indirect immunofluorescence IF for anti-M2, anti-ribosomal protein and anti-LKM1, double immunodiffusion ID for anti-Jo-1 and anti-LKM1, western blotting WB for anti-M2) and with Cyto-Dot HM010. The one hundred and four sera were divided into five groups: Group I (n = 12) with anti-Jo-1 detected by ID; Group II (n = 28) with 26 anti-M2 positive by IF and WB, 2 anti-M2 positive only by WB; Group III (n = 10) with anti-ribosomal protein detected by IF 5 of which precipitated by ID; Group IV (n = 32) with anti-LKM1 by IF and ID divided into 18 AIH2 and 14 HCV; Group V (n = 22) consisting of 14 healthy individuals and 8 patients with hypergammaglobulinemia. Results of this study are similar to those of Cyto-Dot HM010 for the three auto-antibodies already in use. Cyto-Dot 4 is a very good anti-LKM1 confirmation method as it is ID. Copyright John Libbey Eurotext 2003.
Molecular detection of drug resistance in microbes by isotopic techniques: The IAEA experience
International Nuclear Information System (INIS)
Dar, L.; Boussaha, A.; Padhy, A.K.; Khan, B.
2003-01-01
The International Atomic Energy Agency (IAEA) supports various programmes on the uses of radionuclide techniques in the management of human communicable diseases. An important issue, being addressed through several technology transfer projects, is the detection of drug resistance in microbes by radioisotope based molecular-biology diagnostic procedures. The techniques employed include dot blot hybridisation with P-32 labelled oligonucleotide probes to detect point mutations, associated with drug resistance, in microbial genes amplified by the polymerase chain reaction (PCR). Molecular methods have been used for the detection of drug resistance in the malarial parasite, Plasmodium falciparum, and in Mycobacterium tuberculosis. Radioisotope based molecular-biology methods have been demonstrated to have comparative advantages in being sensitive, specific, cost-effective, and suitable for application to large-scale molecular surveillance for drug resistance. (author)
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Luiz Claudio Pereira Ribeiro
2013-09-01
Full Text Available Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1 antibodies (Abs represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP patients. Western blotting (WB for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I and gag (p24 proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.
International Nuclear Information System (INIS)
Bird, T.A.; Gearing, A.J.; Saklatvala, J.
1988-01-01
Functional receptors (IL1-R) for the proinflammatory cytokine interleukin 1 (IL1) were solubilized from plasma membranes of the NOB-1 subclone of murine EL4 6.1 thymoma cells using the zwitterionic detergent 3[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). Membrane extracts were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and ligand blotted with 125 I-labeled recombinant human IL1 alpha in order to reveal proteins capable of specifically binding IL1. A single polydisperse polypeptide of Mr approximately equal to 80,000 was identified in this way, which bound IL1 alpha and IL1 beta with the same affinity as the IL1-R on intact NOB-1 cells (approximately equal to 10(-10) M). The IL1-binding polypeptide was only seen in membranes from IL1-R-bearing cells and did not react with interleukin 2, tumor necrosis factor alpha, or interferon. IL1-R was purified to apparent homogeneity from solubilized NOB-1 membranes by affinity chromatography on wheat germ agglutinin-Sepharose and IL1 alpha-Sepharose. Gel electrophoresis and silver staining of purified preparations revealed a single protein of Mr approximately equal to 80,000 which reacted positively in the ligand-blotting procedure and which we identify as the ligand-binding moiety of the murine IL1-R. Purified IL1-R exhibited the same affinity and specificity as the receptor on intact cells. The relationship of this protein to proteins identified by covalent cross-linking studies is discussed
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Bowlin Gary L
2007-10-01
Full Text Available Abstract Background Electrospinning is a non-mechanical processing strategy that can be used to process a variety of native and synthetic polymers into highly porous materials composed of nano-scale to micron-scale diameter fibers. By nature, electrospun materials exhibit an extensive surface area and highly interconnected pore spaces. In this study we adopted a biological engineering approach to ask how the specific unique advantages of the electrospinning process might be exploited to produce a new class of research/diagnostic tools. Methods The electrospinning properties of nitrocellulose, charged nylon and blends of these materials are characterized. Results Nitrocellulose electrospun from a starting concentration of Conclusion The flexibility afforded by electrospinning process makes it possible to tailor blotting membranes to specific applications. Electrospinning has a variety of potential applications in the clinical diagnostic field of use.
Molecular techniques for MRSA typing: current issues and perspectives
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P. A. Trindade
Full Text Available Staphylococcus aureus has long been recognised as an important pathogen in human disease. Serious staphylococcal infections can frequently occur in inpatients and may lead to dire consequences, especially as to therapy with antimicrobial agents. The increase in the frequency of Methicillin-Resistant Staphylococcus aureus (MRSA as the causal agent of nosocomial infection and the possibility of emergence of resistance to vancomycin demands a quick and trustworthy characterization of isolates and identification of clonal spread within hospitals. Enough information must be generated to permit the implementation of appropriate measures for control of infection, so that outbreaks can be contained. Molecular typing techniques reviewed in this manuscript include: plasmid profile analysis, analysis of chromosomal DNA after enzymatic restriction, Southern blotting, pulsed field gel electrophoresis (PFGE, techniques involving polymerase chain reaction and multilocus sequence typing (MLST. Repetitive DNA Sequence PCR (rep-PCR may be used for screening due to its practicality, low cost and reproducibility. Because of its high discriminatory power Pulsed-Field Gel Electrophoresis (PFGE still remains the gold standard for MRSA typing. New techniques with higher reproducibility and discriminatory power, such as Multi-Locus Sequence Typing (MLST, are appearing. These are mostly useful for global epidemiology studies. Molecular typing techniques are invaluable tools for the assessment of putative MRSA outbreaks and so should be extensively used for this purpose.
Molecular techniques for MRSA typing: current issues and perspectives
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Trindade P. A.
2003-01-01
Full Text Available Staphylococcus aureus has long been recognised as an important pathogen in human disease. Serious staphylococcal infections can frequently occur in inpatients and may lead to dire consequences, especially as to therapy with antimicrobial agents. The increase in the frequency of Methicillin-Resistant Staphylococcus aureus (MRSA as the causal agent of nosocomial infection and the possibility of emergence of resistance to vancomycin demands a quick and trustworthy characterization of isolates and identification of clonal spread within hospitals. Enough information must be generated to permit the implementation of appropriate measures for control of infection, so that outbreaks can be contained. Molecular typing techniques reviewed in this manuscript include: plasmid profile analysis, analysis of chromosomal DNA after enzymatic restriction, Southern blotting, pulsed field gel electrophoresis (PFGE, techniques involving polymerase chain reaction and multilocus sequence typing (MLST. Repetitive DNA Sequence PCR (rep-PCR may be used for screening due to its practicality, low cost and reproducibility. Because of its high discriminatory power Pulsed-Field Gel Electrophoresis (PFGE still remains the gold standard for MRSA typing. New techniques with higher reproducibility and discriminatory power, such as Multi-Locus Sequence Typing (MLST, are appearing. These are mostly useful for global epidemiology studies. Molecular typing techniques are invaluable tools for the assessment of putative MRSA outbreaks and so should be extensively used for this purpose.
2014-01-01
Background It is well known that different Eimeria maxima strains exhibit significant antigenic variation. However, the genetic basis of these phenotypes remains unclear. Methods Total RNA and mRNA were isolated from unsporulated oocysts of E. maxima strains SH and NT, which were found to have significant differences in immunogenicity in our previous research. Two subtractive cDNA libraries were constructed using suppression subtractive hybridization (SSH) and specific genes were further analyzed by dot-blot hybridization and qRT-PCR analysis. Results A total of 561 clones were selected from both cDNA libraries and the length of the inserted fragments was 0.25–1.0 kb. Dot-blot hybridization revealed a total of 86 differentially expressed clones (63 from strain SH and 23 from strain NT). Nucleotide sequencing analysis of these clones revealed ten specific contigs (six from strain SH and four from strain NT). Further analysis found that six contigs from strain SH and three from strain NT shared significant identities with previously reported proteins, and one contig was presumed to be novel. The specific differentially expressed genes were finally verified by RT-PCR and qRT-PCR analyses. Conclusions The data presented here suggest that specific genes identified between the two strains may be important molecules in the immunogenicity of E. maxima that may present potential new drug targets or vaccine candidates for coccidiosis. PMID:24894832
Wang, Hye-young; Kim, Hyunjung; Kim, Yeun; Bang, Hyeeun; Kim, Jong-Pill; Hwang, Joo Hwan; Cho, Sang-Nae; Kim, Tae Ue; Lee, Hyeyoung
2015-10-01
Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531 ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae.
Lin, Wen-Wei; Chen, I-Ju; Cheng, Ta-Chun; Tung, Yi-Ching; Chu, Pei-Yu; Chuang, Chih-Hung; Hsieh, Yuan-Chin; Huang, Chien-Chiao; Wang, Yeng-Tseng; Kao, Chien-Han; Roffler, Steve R; Cheng, Tian-Lu
2016-01-01
Molecular weight markers that can tolerate denaturing conditions and be auto-detected by secondary antibodies offer great efficacy and convenience for Western Blotting. Here, we describe M&R LE protein markers which contain linear epitopes derived from the heavy chain constant regions of mouse and rabbit immunoglobulin G (IgG Fc LE). These markers can be directly recognized and stained by a wide range of anti-mouse and anti-rabbit secondary antibodies. We selected three mouse (M1, M2 and M3) linear IgG1 and three rabbit (R1, R2 and R3) linear IgG heavy chain epitope candidates based on their respective crystal structures. Western blot analysis indicated that M2 and R2 linear epitopes are effectively recognized by anti-mouse and anti-rabbit secondary antibodies, respectively. We fused the M2 and R2 epitopes (M&R LE) and incorporated the polypeptide in a range of 15-120 kDa auto-detecting markers (M&R LE protein marker). The M&R LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the M&R LE protein marker plotted as gel mobility versus the log of the marker molecular weights revealed good linearity with a correlation coefficient R2 value of 0.9965, indicating that the M&R LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected M&R LE protein marker may provide a simple, efficient and economical tool for protein analysis.
International Nuclear Information System (INIS)
Lucher, L.A.; Lego, T.
1989-01-01
We have determined that sodium salicylate, a water-soluble fluor which we use routinely for fluorography with polyacrylamide gels, is also useful for fluorography with thin-layer media. Detection of 3 H-labeled material applied to thin-layer chromatography plates, or nitrocellulose membranes, can be enhanced up to 150-fold after treatment with an aqueous solution of 2 M sodium salicylate, while detection of 35 S-labeled material is enhanced only about 2-fold. We demonstrate the utility of sodium salicylate fluorography in detecting 3H-labeled palmitic acid following thin-layer chromatography and 3 H-labeled proteins following blotting to nitrocellulose
Energy Technology Data Exchange (ETDEWEB)
McMichael, J C; Greisiger, L M; Millman, I [Institute for Cancer Research, Philadelphia, PA (USA). Fox Chase Cancer Center
1981-08-28
Nitrocellulose-protein blotting of serum electrophoresed in agarose gels has been adapted for the study of hepatitis B surface antigen (HBsAg). /sup 125/I-labeled anti-HBs was used as the antigen probe, and the electrophoretic migration was monitored by autoradiography. The method required 3 ..mu..l or less of serum and could detect as little as 1 pg of purified HBsAg. Typically, the authors observed two bands of HBsAg; a moving band which migrated about one-third the distance moved by human serum albumin and a non-migratory band which remained at the loading site. Some examples of the use of the method include: (1) empirical methods for correlating HBsAg concentration in serum to film darkness; (2) observations of mobility changes in serial sera from dialysis patients with chronic HBsAg antigenemia; and (3) detection of related antigens such as antigen from the PLC/PRF/5 hepatoma tissue culture line and the cross-reacting woodchuck hepatitis virus surface antigen (WHsAg).
Techniques for characterization and eradication of potato cyst nematode: a review.
Bairwa, Aarti; Venkatasalam, E P; Sudha, R; Umamaheswari, R; Singh, B P
2017-09-01
Correct identification of species and pathotypes is must for eradication of potato cyst nematodes (PCN). The identification of PCN species after completing the life cycle is very difficult because it is based on morphological and morphometrical characteristics. Genetically different populations of PCN are morphologically same and differentiated based on the host differential study. Later on these traditional techniques have been replaced by biochemical techniques viz, one and two dimensional gel electrophoresis, capillary gel electrophoresis, isozymes, dot blot hybridization and isoelectric focusing etc. to distinguish both the species. One and two dimensional gel electrophoresis has used to examine inter- and intra-specific differences in proteins of Globodera rostochiensis and G. pallida . Now application of PCR and DNA based characterization techniques like RAPD, AFLP and RFLP are the important tools for differentiating inter- and intra specific variation in PCN and has given opportunities to accurate identification of PCN. For managing the PCN, till now we are following integrated pest management (IPM) strategies, however these strategies are not effective to eradicate the PCN. Therefore to eradicate the PCN we need noval management practices like RNAi (RNA interference) or Gene silencing.
DEFF Research Database (Denmark)
Sahm, K.; MacGregor, BJ; Jørgensen, BB
1999-01-01
In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...
Northern blot analysis to investigate the abundance of microorganisms
International Nuclear Information System (INIS)
Krause, D.O.
2005-01-01
areas known as hyper-variable regions which have a high degree of sequence variation. As a result of this structure, it is possible to design signature oligonucleotide probes varying in length from about 15 to 30 nucleotides that are diagnostic of microorganisms at the kingdom, domain, genus and even species level. These signature sequences can be used in a variety of applications such as PCR analysis, construction of clone libraries or direct probing of bulk rRNA. In this chapter, I provide detailed protocols for the analysis of extracted rRNA and give detailed procedures that must be followed to do northern blot analysis of bulk RNA extracted from the rumen
International Nuclear Information System (INIS)
Wang Hui; Yi Xuping; Hu Bingqiang; Zeng Liang; Liu Yisong; Liang Songping
2007-01-01
Objective: To examine the variation of protein expression in nasopharyngeal carcinoma cell lines with different biological characteristics and to identify the radiobiological associated proteins. Methods: Biological characteristics of 5-8F and 6-10B were compared by flow cytometry assay after irradiation. The total proteins of 5-8F and 6-10B were separated by immobilized pH gradient(IPG) IEF-SDS two-dimensional gel eleetrophoresis technique. The differentially expressed proteins were cut from the gel and digested into peptides for MALDI-TOF MS and the Q-TOF mass spectrometric analysis. Identification of protein was made through searching in protein sequence database. Protein expressions were examined by western blot and immunohistochemistry method. Results: Nine most differentially expressed proteins between 5-8F cell and 6-10B cell were identified, p73 and CK19 expression examined by western blot were conformal with that by proteomic method, p73 expression in 5-8F cell was higher than in 6-10B cell. CK19 expression in 6- 10B cell was higher than in 5-8F cell. Conclusion: Differentially expression of proteins exist in nasopharyngeal carcinoma cell lines with different biological characteristics. These proteins may be associated with cell radiobiological characteristic with the p73 as a potential biomarker. (authors)
Aisa, M J; Castillejo, S; Gallego, M; Fisa, R; Riera, M C; de Colmenares, M; Torras, S; Roura, X; Sentis, J; Portus, M
1998-02-01
Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.
Blot hybridization analysis of TCR genes of T cells for five people exposed in a radiation accident
International Nuclear Information System (INIS)
Min Rui; Liu Benti; Cheng Tianmin; Yang Rujun; Meng Xiangshun; Xiao Jinsong
1996-01-01
Human lymphocyte total DNA was prepared in agarose plug by mixing cells with low melting agarose, and two restriction endonucleases were used for digestion of the total DNA with human α and β TCR cDNA probes. The total digested DNA from five people who were whole body exposed to 2.0-2.5 Gy ionizing radiation in an accident 4.5 years ago was hybridized by Southern blot method. The results showed that no obvious difference in hybridization bands was found between controls and the five victims when hybridizations were fulfilled in the total DNA which was digested by Hind III restriction endonuclease with both α and β probes. However, when the total DNA was digested with restriction endonuclease EcoR I and was hybridized with TCR α probe, four of the five exposed people showed a different hybridizing band pattern compared with the controls. The results are also discussed
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Sarada Subramanian
2016-01-01
Full Text Available Background and Purpose: Definitive diagnosis of Creutzfeldt–Jakob disease (CJD requires demonstration of infective prion protein (PrPSc in brain tissues by immunohistochemistry or immunoblot, making antemortem diagnosis of CJD difficult. The World Health Organization (WHO recommends detection of 14-3-3 protein in cerebrospinal fluid (CSF in cases of dementia, with clinical correlation, as a useful diagnostic marker for CJD, obviating the need for brain biopsy.This facility is currently available in only a few specialized centers in the West and no commercial kit is available for clinical diagnostic use in India. Hence the objective of this study was to develop an in-house sensitive assay for quantitation of 14-3-3 protein and to evaluate its diagnostic potential to detect 14-3-3 proteins in CSF as a biomarker in suspected cases of CJD. Materials and Methods: A minigene expressing the “core” 14-3-3 protein was synthesized by overlapping polymerase chain reaction (PCR and the recombinant protein was produced by employing a bacterial expression system. Polyclonal antibodies raised in rabbit against the purified recombinant protein were used for developing a dot blot assay with avidin-biotin technology for signal amplification and quantitation of 14-3-3 protein in CSF. Results: The results in the present study suggest the diagnostic potential of the dot blot method with about 10-fold difference (P< 0.001 in the CSF levels of 14-3-3 protein between the CJD cases (N= 50 and disease controls (N= 70. The receiver operating characteristic (ROC analysis of the results suggested an optimal cutoff value of 2 ng/mL. Conclusions: We have developed an indigenous, economical, and sensitive dot blot method for the quantitation of 14-3-3 protein in CSF.
Molecular biology - Part I: Techniques, terminology, and concepts
International Nuclear Information System (INIS)
Brown, J. Martin
1996-01-01
Purpose/Objective: One of the barriers to understanding modern molecular biology is the lack of a clear understanding of the relevant terminology, techniques, and concepts. This refresher course is intended to address these deficiencies starting from a basic level. The lecture will cover many of the common uses of recombinant DNA, including gene cloning and manipulation. The goal is to enable the nonspecialist to increase his or her understanding of molecular biology in order to more fully enjoy reading current publications and/or listening seminars. Radiation biologists trying to understand a little more molecular biology should also benefit. The following concepts will be among those explained and illustrated: restriction endonucleases, gel electrophoresis, gene cloning, use of vectors such as plasmids, bacteriophage, cosmids and viruses, cDNA and genomic libraries, Southern, Northern, and Western blotting, fluorescent in situ hybridization, polymerase chain reaction (PCR), gel retardation, and reporter gene assays
Lopez-Jimena, B; Cherif, N; Garcia-Rosado, E; Infante, C; Cano, I; Castro, D; Hammami, S; Borrego, J J; Alonso, M C
2010-10-01
To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46.87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90.62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56.25%). This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.
Scott, M C; Harmon, J R; Tsao, J I; Jones, C J; Hickling, G J
2012-05-01
Determining the host preference of vector ticks is vital to elucidating the eco-epidemiology of the diseases they spread. Detachment of ticks from captured hosts can provide evidence of feeding on those host species, but only for those species that are feasible to capture. Recently developed, highly sensitive molecular assays show great promise in allowing host selection to be determined from minute traces of host DNA that persist in recently molted ticks. Using methods developed in Europe as a starting-point, we designed 12S rDNA mitochondrial gene probes suitable for use in a reverse line blot (RLB) assay of ticks feeding on common host species in the eastern United States. This is the first study to use the 12S mitochondrial gene in a RLB bloodmeal assay in North America. The assay combines conventional PCR with a biotin-labeled primer and reverse line blots that can be stripped and rehybridized up to 20 times, making the method less expensive and more straightforward to interpret than previous methods of tick bloodmeal identification. Probes were designed that target the species, genus, genus group, family, order, or class of eight reptile, 13 birds, and 32 mammal hosts. After optimization, the RLB assay correctly identified the current hostspecies for 99% of ticks [Amblyomma americanum (L.) and eight other ixodid tick species] collected directly from known hosts. The method identified previous-host DNA for approximately half of all questing ticks assayed. Multiple bloodmeal determinations were obtained in some instances from feeding and questing ticks; this pattern is consistent with previous RLB studies but requires further investigation. Development of this probe library, suitable for eastern U.S. ecosystems, opens new avenues for eco-epidemiological investigations of this region's tick-host systems.
Retrospective study of hemoparasites in cattle in southern Italy by reverse line blot hybridization.
Ceci, Luigi; Iarussi, Fabrizio; Greco, Beatrice; Lacinio, Rosanna; Fornelli, Stefania; Carelli, Grazia
2014-06-01
Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied.
Antibody responses to Borrelia burgdorferi detected by western blot vary geographically in Canada.
Ogden, Nicholas H; Arsenault, Julie; Hatchette, Todd F; Mechai, Samir; Lindsay, L Robbin
2017-01-01
Lyme disease is emerging in eastern and central Canada, and most cases are diagnosed using the two-tier serological test (Enzyme Immuno Assay [EIA] followed by Western blot [WB]). Simplification of this algorithm would be advantageous unless it impacts test performance. In this study, accuracy of individual proteins of the IgG WB algorithm in predicting the overall test result in samples from Canadians was assessed. Because Borrelia burgdorferi strains vary geographically in Canada, geographic variations in serological responses were also explored. Metrics of relative sensitivity, specificity and the kappa statistic measure of concordance were used to assess the capacity of responses to individual proteins to predict the overall IgG WB result of 2524 EIA (C6)-positive samples from across Canada. Geographic and interannual variations in proportions of samples testing positive were explored by logistic regression. No one protein was highly concordant with the IgG WB result. Significant variations were found amongst years and geographic regions in the prevalence of samples testing positive using the overall IgG WB algorithm, and for individual proteins of the algorithm. In most cases the prevalence of samples testing positive were highest in Nova Scotia, and lower in samples from Manitoba westwards. These findings suggest that the current two tier test may not be simplified and continued use of the current two-tier test method and interpretation is recommended. Geographic and interannual variations in the prevalence of samples testing positive may be consistent with B. burgdorferi strain variation in Canada, and further studies are needed to explore this.
Antibody responses to Borrelia burgdorferi detected by western blot vary geographically in Canada.
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Nicholas H Ogden
Full Text Available Lyme disease is emerging in eastern and central Canada, and most cases are diagnosed using the two-tier serological test (Enzyme Immuno Assay [EIA] followed by Western blot [WB]. Simplification of this algorithm would be advantageous unless it impacts test performance. In this study, accuracy of individual proteins of the IgG WB algorithm in predicting the overall test result in samples from Canadians was assessed. Because Borrelia burgdorferi strains vary geographically in Canada, geographic variations in serological responses were also explored. Metrics of relative sensitivity, specificity and the kappa statistic measure of concordance were used to assess the capacity of responses to individual proteins to predict the overall IgG WB result of 2524 EIA (C6-positive samples from across Canada. Geographic and interannual variations in proportions of samples testing positive were explored by logistic regression. No one protein was highly concordant with the IgG WB result. Significant variations were found amongst years and geographic regions in the prevalence of samples testing positive using the overall IgG WB algorithm, and for individual proteins of the algorithm. In most cases the prevalence of samples testing positive were highest in Nova Scotia, and lower in samples from Manitoba westwards. These findings suggest that the current two tier test may not be simplified and continued use of the current two-tier test method and interpretation is recommended. Geographic and interannual variations in the prevalence of samples testing positive may be consistent with B. burgdorferi strain variation in Canada, and further studies are needed to explore this.
Bartholomew, J.L.; Smith, C.E.; Rohovec, J.S.; Fryer, J.L.
1989-01-01
The tissue response of Salmo gairdneri Richardson, against the myxosporean parasite. Ceratomyxa shasta (Noble), was investigated using histological techniques, scanning electron microscopy and immunological methods. The progress of infection in C. shasta-susceptible and resistant steelhead and rainbow trout was examined by standard histological techniques and by indirect fluorescent antibody methods using monoclonal antibodies directed against C. shasta antigens. Trophozoite stages were first observed in the posterior intestine and there was indication that resistance was due to the inability of the parasite to penetrate this tissue rather than to an inflammatory response. Examination of a severely infected intestine by scanning electron microscopy showed extensive destruction of the mucosal folds of the posterior intestine. Western blotting and indirect fluorescent antibody techniques were used to investigate the immunological component of the host response. No antibodies specific for C. shasta were detected by either method.
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Osmir Cabrera-Blanco
2017-04-01
Full Text Available Las autoridades regulatorias recomiendan el uso de técnicas de Resonancia Magnética Nuclear o técnicas serológicas para la determinación de la identidad de los antígenos presentes en las vacunas conjugadas. Con la aparición de las vacunas conjugadas multivalentes, se ha hecho necesario recurrir a técnicas inmunoquímicas con la utilización de anticuerpos monoclonales para aumentar la sensibilidad en la determinación de la identidad de los antígenos en dichas vacunas conjugadas. El objetivo del presente trabajo fue establecer las condiciones óptimas de trabajo que permitieran utilizar la técnica del Dot Blot para determinar la identidad de los antígenos en vacunas conjugadas de Streptococcus pneumoniae serotipo 19F. Para ello se estudiaron los tiempos de incubación, la influencia del reactivo en la solución de bloqueo; también las concentraciones óptimas del anticuerpo monoclonal y de los ingredientes farmacéuticos activos, así como los volúmenes de aplicación óptimos para estos y vacunas. Se utilizó un anticuerpo monoclonal contra el polisacárido capsular del serotipo 19F de neumococo. Las muestras empleadas en este trabajo fueron lotes de ingredientes farmacéuticos activos de conjugados de polisacárido capsular 19F y lotes de un candidato vacunal cubano conjugado heptavalente contra neumococos. Los resultados mostraron que para la determinación de la identidad antigénica fueron suficientes 10 µL de muestras de los principios activos a una concentración de 125 µg/mL e igual volumen para las vacunas heptavalentes. Quedó demostrado que una concentración de 1 µg/mL para el anticuerpo monoclonal y tiempos de incubación de 30 min a 37 °C fueron suficientes para la determinación. Estos resultados permiten concluir que quedaron establecidas las condiciones óptimas de trabajo para determinar la identidad antigénica por Dot Blot del polisacárido capsular de S. pneumoniae serotipo 19F presente en las vacunas
Traversa, Donato; Iorio, Raffaella; Klei, Thomas R.; Kharchenko, Vitaliy A.; Gawor, Jakub; Otranto, Domenico; Sparagano, Olivier A. E.
2007-01-01
The ability of a reverse line blot (RLB) assay to identify 13 common species of equine small strongyles (cyathostomins) and to discriminate them from three Strongylus spp. (large strongyles) was demonstrated. The assay relied on the specific hybridization of PCR-amplified intergenic spacer DNA fragments of the nuclear ribosomal DNA to membrane-bound species-specific probes. All cyathostomins examined were unequivocally identified and simultaneously discriminated from each other and from three large strongyles (Strongylus edentatus, Strongylus equinus, and Strongylus vulgaris). This assay will enable the accurate and rapid identification of equine cyathostomins irrespective of their life cycle stage, opening important avenues for a better understanding of their biology and epidemiology and of the pathogenesis of cyathostomin-associated disease. In particular, this RLB method promises to be a powerful diagnostic tool to determine the roles of individual species in the pathogenesis of mixed infections and to elucidate some aspects of cyathostominosis. Also, it could represent a basic step toward the development of a rapid and simple molecular test for the early detection of drug-resistant genotypes of horse strongyle species. PMID:17626168
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William H Roldán
2009-05-01
Full Text Available To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis. Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%, whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.
International Nuclear Information System (INIS)
Bu Hengfu; Chen Juan; Shen Rongsen; Ma Liren; Xu Yongqiang
1996-01-01
Southern blotting hybridization (SBH) using a 32 P-labeled probe is one of the most practical methods for genetic diagnosis of pathogen. On the basis of establishing PCR and nested PCR for detecting human cytomegalovirus (HCMV), a 32 P-labeled probe was prepared with the amplified products of 613 bp PCR outer primers and hybridized with 300 bp inner primer amplified product, resulting in increase in detecting sensitivity from 17 ng (in 1.2% agarose electrophoresis) before SBH to 500 pg (autoradiographed), in other words, increasing the sensitivity of detecting HCMV by 10 2 dilutions after using SBH. The method of PCR and SBH using a 32 P-labeled probe could detect less than 1 gene copy of HCMV, therefore, it is a rapid and reliable diagnosis method for detecting HCMV latent infection
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Eikan Mishima
Full Text Available The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A, N6-methyladenosine (m6A, pseudouridine, and 5-methylcytidine (m5C showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism.
Mishima, Eikan; Jinno, Daisuke; Akiyama, Yasutoshi; Itoh, Kunihiko; Nankumo, Shinnosuke; Shima, Hisato; Kikuchi, Koichi; Takeuchi, Yoichi; Elkordy, Alaa; Suzuki, Takehiro; Niizuma, Kuniyasu; Ito, Sadayoshi; Tomioka, Yoshihisa; Abe, Takaaki
2015-01-01
The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism.
Muro, Claudio; Gomez-Puerta, Luis A; Flecker, Robert H; Gamboa, Ricardo; Barreto, Percy Vilchez; Dorny, Pierre; Tsang, Victor C W; Gilman, Robert H; Gonzalez, Armando E; Garcia, Hector H; O'Neal, Seth E; For The Cysticercosis Working Group In Peru
2017-12-01
The lentil lectin glycoprotein enzyme-linked immunoelectrotransfer blot (LLGP EITB, reported sensitivity 99% and specificity 100%) is used as a serologic marker of exposure to Taenia solium in pigs. However, only a limited number of parasites have been evaluated for cross reactivity. Pigs may host other related cestode infections, including Taenia hydatigena, which have not been formally evaluated for cross-reactions. We investigated a corral in Tumbes, Peru, a region where a cysticercosis elimination demonstration project was completed in 2012. In this corral, 14/19 (73.7%) 6-8-week-old piglets were reactive to GP50 on LLGP EITB, and all had circulating Taenia sp. antigens. From eight necropsied piglets; four were infected with T. hydatigena metacestodes whereas none had evidence of T. solium infection. Two resident dogs were subsequently confirmed to have T. hydatigena taeniasis. These results suggest GP50 cross-reactivity in T. hydatigena- infected pigs, although controlled experimental infection is needed to confirm this hypothesis.
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Kelly Davelois
Full Text Available Objetivo. Determinar el rendimiento diagnóstico de la técnica de Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana. Materiales y métodos. Estudio transversal de evaluación de prueba diagnóstica. Se obtuvieron los antígenos de excreción-secreción de las larvas de Taenia solium, quistes de Echinococcus granulosus; y la forma adulta de Fasciola hepática; que luego fueron separados electroforéticamente en geles de poliacrilamida individuales, transferidos y fijados a una membrana de nitrocelulosa para ser enfrentados con sueros de pacientes con las tres parasitosis. La sensibilidad de la técnica se evaluó empleando 300 sueros individuales, 60 pools de dos parasitosis y 20 pools de tres parasitosis y la especificidad con 75 sueros de pacientes con otras parasitosis, 10 de pacientes con otras enfermedades y 15 sueros de personas no parasitadas. Resultados. La técnica reconoció trece glicoproteínas (GP: GP 35, 31, 24, 23, 18, 17, 14 y 13 kDa para cisticercosis, GP 8,16 y 21 kDa para hidatidosis y GP: 17 y 23 kDa para fascioliasis. La prueba detectó la presencia de anticuerpos alcanzando una sensibilidad de 96% (IC95%: 94,62-98,54% en la detección de una o las trece bandas, una especificidad de 100% (IC95%: 99,50 - 100,00%; individualmente, se tuvo una sensibilidad para cisticercosis de 97% (IC95%: 93,16-100%, para hidatidosis de 94% (IC95%: 88,85-99,15% y para fascioliasis de 96% (IC95%: 91,66-100%. Conclusiones. La prueba de Western blot es eficaz en la detección, simultanea de anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana, y puede ser utilizada como prueba de descarte o confirmatoria en zonas endémicas.
International Nuclear Information System (INIS)
Maria Lina R; Andi Yasmon
2011-01-01
There are many commercial ELISA and rapid test kits that have been used for blood screening; however, the kits can give false positive and negative results. Therefore, RT-PCR (Reverse Transcription Polymerase Chain Reaction) - Dot Blot Hybridization based on radioisotope 32 P (RDBR) method was developed in this research, to compare the method with the conventional RT-PCR and commercial ELISA Enzyme-Linked lmmunosorbent Assay) kit. This method is efficient for screening of large blood specimens and surveillance study. Eighty seven samples were used and serum of the samples were tested by ELISA to detect HIV-1. The HIV-l RNA genome was extracted from plasma samples and tested using the RT-PCR and RDBR methods. Of 87 samples that were tested, the rates of positive testing of the RT-PCR, the RDBR, and the ELISA were 71.26%, 74.71%, and 80.46%, respectively. The RDBR (a combination of RTPCR and dot blot hybridization) was more sensitive than conventional RT-PCR by showing 3.45% in increase number of positive specimens. The results showed that of 9 samples (10.34%) were negative RDBR and positive ELISA, while 4 samples (4.60%) were negative ELISA and positive RDBR. The two methods showed slightly difference in the results but further validation is still needed. However, RDBR has high potential as an alternative method for screening of blood in large quantities when compared to method of conventional RT-PCR and ELISA. (author)
DEFF Research Database (Denmark)
Sahm, K.; MacGregor, BJ; Jørgensen, BB
1999-01-01
In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization...... between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulpho-bacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...
Methogo, Ruth Menque; Dansette, Patrick M.; Klarskov, Klaus
2007-12-01
A combined approach based on two-dimensional electrophoresis-immuno-blotting and nanoliquid chromatography coupled on-line with electrospray ionization mass spectrometry (nLC-MS/MS) was used to identify proteins modified by a reactive intermediate of tienilic acid (TA). Liver homogenates from rats exposed to TA were fractionated using ultra centrifugation; four fractions were obtained and subjected to 2D electrophoresis. Following transfer to PVDF membranes, modified proteins were visualized after India ink staining, using an anti-serum raised against TA and ECL detection. Immuno-reactive spots were localized on the PVDF membrane by superposition of the ECL image, protein spots of interest were excised, digested on the membrane with trypsin followed by nLC-MS/MS analysis and protein identification. A total of 15 proteins were identified as likely targets modified by a TA reactive metabolite. These include selenium binding protein 2, senescence marker protein SMP-30, adenosine kinase, Acy1 protein, adenosylhomocysteinase, capping protein (actin filament), protein disulfide isomerase, fumarylacetoacetase, arginase chain A, ketohexokinase, proteasome endopeptidase complex, triosephosphate isomerase, superoxide dismutase, dna-type molecular chaperone hsc73 and malate dehydrogenase.
Kilic, Tugba; Erdem, Arzum; Ozsoz, Mehmet; Carrara, Sandro
2018-01-15
As being the most extensively studied, non-coding, evolutionary conserved, post-transcriptional gene regulators of genome, microRNAs (miRNAs) have taken great attention among various disciplines due to their important roles in biological processes and link with cancer. Due to their diagnostic value, there have been many conventional methods used in detection of miRNAs including northern blotting, quantitative real time PCR (qRT-PCR) and microarray technology besides novel techniques based on various nanotechnology approaches and molecular biology tools including miRNA biosensors. The aim of this review is to explain the importance of miRNAs in biomedical field with an emphasis on early cancer diagnosis by overviewing both research based and commercially available miRNA detection methods in the last decade considering their strengths and weakness with an emphasis on miRNA biosensors. Copyright © 2017 Elsevier B.V. All rights reserved.
Exploration of the speleotherapeutic potential through the cellular and molecular biology techniques
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Munteanu Constantin
2011-02-01
Full Text Available Objective: Exploring the speleotherapy effects on morphology and physiology of dermal and pulmonary fibroblast obtained from Wistar rats tissue in normal conditions and after induction of experimental “astma” awareness with ovalbumin. Materials and methods: Before initiation of dermal and pulmonary fibroblast cultures, 60 of Wistar rats 75-100 g were divided into two groups: control and sensitized with ovalalbumin. 10 animals of each group were sent to Cacica and Dej salt mines and maintained in a speleotherapy regime. Another 10 animals in each group were monitored separately in INRMFB Biobase . Dermal and pulmonary fibroblast cultures were initiated by enzymatic techniques from appropriate tissue taken of each group Wistar rats. Morphological monitoring was done by phase contast microscopy; biochemical and molecular changes of cultures obtained from animals treated speleothropic compared to control, was experimental establised by electrophoresis and Western Blotting techniques.Results: Experimental data revealed the expression of several proteins after the speleotherapeutic treatment. These data were analysed compared with control, using a specific software.Conclusions: Speleotherapeutic treatment of Wistar rats caused significant differences in morphology and protein expression of dermal and pulmonary fibroblatst grown in the laboratory. These differences support the protective effects of speleotherapy compared with data obtained from animals untreated and sensitized with ovalbumin, having induced experimental asthma status.
Yamamoto, Norio; Yamashita, Yoko; Yoshioka, Yasukiyo; Nishiumi, Shin; Ashida, Hitoshi
2016-08-01
Membrane proteins account for 70% to 80% of all pharmaceutical targets, indicating their clinical relevance and underscoring the importance of identifying differentially expressed membrane proteins that reflect distinct disease properties. The translocation of proteins from the bulk of the cytosol to the plasma membrane is a critical step in the transfer of information from membrane-embedded receptors or transporters to the cell interior. To understand how membrane proteins work, it is important to separate the membrane fraction of cells. This unit provides a protocol for rapidly obtaining plasma membrane fractions for western blot analysis. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.
Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth
2018-02-01
Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.
International Nuclear Information System (INIS)
Ravanshad, M.; Sabahi, F.; Mahboudi, F.; Sabahi, F.
2006-01-01
The Western blot (WB) assay is the most widely accepted confirmatory assay for the detection and confirmation of antibodies to human immunodeficiency virus type 1 (HIV-1) and 2 (HIV-2). However, indeterminate WB reactivity to HIV-1 and HIV-2 proteins may occur in individuals who do not appear to be infected with HIV. In this study, we describe the results of indeterminate WB reactivity in Iranian patients with discordant screening assays. The samples were obtained from Iranian Blood Transfusion Center, Tehran, Iran and evaluated in the Biotechnology Process Development Center, Pasteur Institute of Iran, Tehran, Iran between 2003 and 2004. A total of 4707 were tested for the presence of HIV-1 antibodies. Six hundred and four (12.8%) patients tested for HIV were positive for HIV-1 antibody. Nine (1.49%) have discordant results among screening assays and indeterminate WB results as interpreted by Centers for Disease Control and Prevention (CDC) criteria. Most (66.7%) of these indeterminate WB results were due to p24 reactivity. However, 2(22.2%) display reactivity to both gp41 and gp120 proteins [Positive by World Health Organization (WHO) criteria]. Of 9 WB assays initially indeterminate by the CDC criteria and with follow-up samples 8(88.8%) became negative when retested subsequently while one (11.1%) remained indeterminate for more than a year and were thus considered negative. In addition all the indeterminate samples were negative when assessed by polymerase chain reaction assay. In general, there were was an 88.8% concordance between the CDC and WHO criteria for an indeterminate WB result. The CDC II criteria for an indeterminate WB result. The CDC II criteria best met the specified objectives for diagnosis in our setting. (author)
Bamman, M. M.; Clarke, M. S.; Talmadge, R. J.; Feeback, D. L.
1999-01-01
Talmadge and Roy (J. Appl. Physiol. 1993, 75, 2337-2340) previously established a sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) protocol for separating all four rat skeletal muscle myosin heavy chain (MHC) isoforms (MHC I, IIa, IIx, IIb); however, when applied to human muscle, the type II MHC isoforms (Ila, IIx) are not clearly distinguished. In this brief paper we describe a modification of the SDS-PAGE protocol which yields distinct and consistent separation of all three adult human MHC isoforms (MHC I, IIa, IIx) in a minigel system. MHC specificity of each band was confirmed by Western blot using three monoclonal IgG antibodies (mAbs) immunoreactive against MHCI (mAb MHCs, Novacastra Laboratories), MHCI+IIa (mAb BF-35), and MHCIIa+IIx (mAb SC-71). Results provide a valuable SDS-PAGE minigel technique for separating MHC isoforms in human muscle without the difficult task of casting gradient gels.
Jean, J; Blais, B; Darveau, A; Fliss, I
2001-12-01
A nucleic acid sequence-based amplification (NASBA) technique for the detection of hepatitis A virus (HAV) in foods was developed and compared to the traditional reverse transcription (RT)-PCR technique. Oligonucleotide primers targeting the VP1 and VP2 genes encoding the major HAV capsid proteins were used for the amplification of viral RNA in an isothermal process resulting in the accumulation of RNA amplicons. Amplicons were detected by hybridization with a digoxigenin-labeled oligonucleotide probe in a dot blot assay format. Using the NASBA, as little as 0.4 ng of target RNA/ml was detected per comparison to 4 ng/ml for RT-PCR. When crude HAV viral lysate was used, a detection limit of 2 PFU (4 x 10(2) PFU/ml) was obtained with NASBA, compared to 50 PFU (1 x 10(4) PFU/ml) obtained with RT-PCR. No interference was encountered in the amplification of HAV RNA in the presence of excess nontarget RNA or DNA. The NASBA system successfully detected HAV recovered from experimentally inoculated samples of waste water, lettuce, and blueberries. Compared to RT-PCR and other amplification techniques, the NASBA system offers several advantages in terms of sensitivity, rapidity, and simplicity. This technique should be readily adaptable for detection of other RNA viruses in both foods and clinical samples.
Cernanská, Dana; Paoletti, Barbara; Králová-Hromadová, Ivica; Iorio, Raffaella; Cudeková, Patrícia; Milillo, Piermarino; Traversa, Donato
2009-03-09
Five horse farms located in eastern Slovakia were investigated for the presence of benzimidazole-resistant strongyles by faecal egg count reduction test and egg hatch assay. Coprocultures were prepared for each farm from faecal samples taken pre- and post-treatment and harvested larvae were molecularly examined with a Reverse Line Blot assay. Faecal egg count reduction values ranged from 0 to 52.5% and all farms were positive for benzimidazole-resistant cyathostomins. Seven benzimidazole-resistant cyathostomin species were molecularly identified on farms before and also after treatment. These data demonstrate that resistance to benzimidazoles is well established in cyathostomin populations from horse farms in the Slovak Republic and that the molecular assay was able to determine the species-specific distribution of resistant cyathostomins under field conditions.
DEFF Research Database (Denmark)
Jantzen, Christian; Østergaard, Per
2006-01-01
Med udgangspunkt i en række cases på vellykkede oplevelsesøkonomiske forretningsmodeller argumenterer artiklen for, at oplevelsesprodukter skal bygge på et klart tema, som forbrugeren kan koble sig sanse- og følelsesmæssigt op på. Forbrugeren skal kunne omsætte produktets pirringer til egne erfar...
DEFF Research Database (Denmark)
Leroyer, Patrick
2010-01-01
The purpose of this article is to reassert the crucial importance of access to data in lexicographic information tools and, expanding on this, to establish the existence of two distinct lexicographic access modes - consultation and navigation. It is explained how the tools can be decalibrated whe...... balance between user, access, and data is disturbed, and how access to data then is jeopardized. Taking online wine guides as a case in point, it is shown how such multifunctional information tools do benefit from a lexicographic design featuring both access modes....
Silva, M G; Marques, P X; Oliva, A
2010-12-15
Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR-reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases. Copyright © 2010 Elsevier B.V. All rights reserved.
International Nuclear Information System (INIS)
Stolf, A.M.S.; Umezawa, E.S.; Zingales, B.
1990-01-01
A radioactive Western blotting technique was developed by which the reactivity of Immunoglobulins (IGs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse. T.cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgC specific antibodies. A different pattern of reactivity with acute Chagas disease patients sera was thus obtained. (author)
Branda, John A; Aguero-Rosenfeld, Maria E; Ferraro, Mary Jane; Johnson, Barbara J B; Wormser, Gary P; Steere, Allen C
2010-01-01
Standard 2-tiered immunoglobulin G (IgG) testing has performed well in late Lyme disease (LD), but IgM testing early in the illness has been problematic. IgG VlsE antibody testing, by itself, improves early sensitivity, but may lower specificity. We studied whether elements of the 2 approaches could be combined to produce a second-tier IgG blot that performs well throughout the infection. Separate serum sets from LD patients and control subjects were tested independently at 2 medical centers using whole-cell enzyme immunoassays and IgM and IgG immunoblots, with recombinant VlsE added to the IgG blots. The results from both centers were combined, and a new second-tier IgG algorithm was developed. With standard 2-tiered IgM and IgG testing, 31% of patients with active erythema migrans (stage 1), 63% of those with acute neuroborreliosis or carditis (stage 2), and 100% of those with arthritis or late neurologic involvement (stage 3) had positive results. Using new IgG criteria, in which only the VlsE band was scored as a second-tier test among patients with early LD (stage 1 or 2) and 5 of 11 IgG bands were required in those with stage 3 LD, 34% of patients with stage 1, 96% of those with stage 2, and 100% of those with stage 3 infection had positive responses. Both new and standard testing achieved 100% specificity. Compared with standard IgM and IgG testing, the new IgG algorithm (with VlsE band) eliminates the need for IgM testing; it provides comparable or better sensitivity, and it maintains high specificity.
Arroyo, Gianfranco; Rodriguez, Silvia; Lescano, Andres G; Alroy, Karen A; Bustos, Javier A; Santivañez, Saul; Gonzales, Isidro; Saavedra, Herbert; Pretell, E Javier; Gonzalez, Armando E; Gilman, Robert H; Tsang, Victor C W; Garcia, Hector H
2018-01-06
The enzyme-linked immunoelectrotransfer blot (EITB) assay is the reference serological test for neurocysticercosis (NCC). A positive result on EITB does not always correlate with the presence of active infections in the central nervous system (CNS), and patients with a single viable brain cyst may be EITB negative. Nonetheless, EITB antibody banding patterns appears to be related with the expression of 3 protein families of Taenia solium, and in turn with the characteristics of NCC in the CNS (type, stage, and burden of viable cysts). We evaluated EITB antibody banding patterns and brain imaging findings of 548 NCC cases. Similar banding patterns were grouped into homogeneous classes using latent class analysis. The association between classes and brain imaging findings was assessed. Four classes were identified. Class 1 (patients negative or only positive to the GP50 band, related to the protein family of the same name) was associated with nonviable or single viable parenchymal cysticerci; class 2 (patients positive to bands GP42-39 and GP24, related to the T24-42 protein family, with or without anti-GP50 antibodies) was associated with intraparenchymal viable and nonviable infections; classes 3 and 4 (positive to GP50, GP42-39, and GP24 but also responding to low molecular weight bands GP21, GP18, GP14, and GP13, related to the 8 kDa protein family) were associated with extraparenchymal and intraparenchymal multiple viable cysticerci. EITB antibody banding patterns correlate with brain imaging findings and complement imaging information for the diagnosis of NCC and for staging NCC patients. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.
Directory of Open Access Journals (Sweden)
CARNEIRO-PROIETTI A.B.F.
1999-01-01
Full Text Available The clinical and public health importance of indeterminate results in HIV-1/2 testing is still difficult to evaluate in volunteer blood donors. At Fundação Hemominas, HIV-1/2 ELISA is used as the screening test and, if reactive, is followed by Western blot (WB. We have evaluated 84 blood donors who had repeatedly reactive ELISA tests for HIV-1/2, but indeterminate WB results. Sixteen of the 84 donors (19.0% had history of sexually transmitted diseases; 18/84 (21.4% informed receiving or paying for sex; 3/84 (3.6% had homosexual contact; 2/26 women (7.6% had past history of multiple illegal abortions and 3/84 (3.6% had been previously transfused. Four out of 62 donors (6.5% had positive anti-nuclear factor (Hep2, with titles up to 1:640. Parasitological examination of the stool revealed eggs of S. mansoni in 4/62 (6.4% donors and other parasites in 8/62 (12.9%. Five (5.9% of the subjects presented overt seroconversion for HIV-1/2, 43/84 (51.2% had negative results on the last visit, while 36/84 (42.9% remained WB indeterminate. Although some conditions could be found associated with the HIV-1/2 indeterminate WB results and many donors had past of risky behavior, the significance of the majority of the results remains to be determined.
Walpurgis, Katja; Thomas, Andreas; Schänzer, Wilhelm; Thevis, Mario
2016-02-01
Myostatin is a key regulator of skeletal muscle growth and inhibition of its signaling pathway results in an increased muscle mass and function. The aim of this study was to develop a qualitative detection assay for myostatin-neutralizing antibodies for doping control purposes by using immunological approaches. To detect different types of myostatin-neutralizing antibodies irrespective of their amino acid sequence, an immunological assay specific for antibodies directed against myostatin and having a human Fc domain was established. Affinity purification and Western blotting strategies were combined to allow extracting and identifying relevant analytes from 200 μL of plasma/serum in a non-targeted approach. The assay was characterized regarding specificity, linearity, precision, robustness, and recovery. The assay was found to be highly specific, robust, and linear from 0.1 to 1 μg/mL. The precision was successfully specified at three different concentrations and the recovery of the affinity purification was 58%. Within this study, an immunological detection assay for myostatin-neutralizing antibodies present in plasma/serum specimens was developed and successfully characterized. The presented approach can easily be modified to include other therapeutic antibodies and serves as proof-of-concept for the detection of antibody-based myostatin inhibitors in doping control samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Seyyed Javad SEYYEDTABAEI
2017-06-01
Full Text Available Background: Visceral leishmaniasis (VL and cutaneous leishmaniasis (CL are important public health problems in Iran. We aimed to evaluate the diagnostic potential of Western blot (WB compared with indirect immunofluorescence test (IFAT to serodiagnosis of leishmaniasis.Methods: This study was performed from 2010-2014 and participants were different parts of Iran. Serum samples were obtained from 43 patients with proven CL, 33 patients with proven VL, 39 patients with other parasitic diseases and 23 healthy individuals. Results: WB sensitivity for CL and VL was 100% and 91%, compared to IFA 4.6% and 87.8%, respectively. Sera from patients with CL and VL recognized numerous antigens with molecular weights ranging from 14 to 68 kDa and 12 to 94 kDa, respectively. The most sensitive antigens were 14 and 16 kDa for CL recognized by 100% of the sera from patients with proven CL and 12, 14 and 16 kDa for VL, recognized by 63.6%, 100% and 63.6% of the sera from patients with proven VL respectively. WB analysis is more sensitive than IFAT for the diagnosis of leishmaniasis particularly in cases of cutaneous leishmaniasis. The 12, 14 and 16 kDa can be valuable diagnostic molecules for serodiagnosis of leishmaniasis because at least two immunogenic molecules were simultaneously detected by all patient sera, as well as produced antibodies against these antigens have no cross-reactivity with other control groups.Conclusion: WB could be useful for screening and serodiagnosis of CL and VL in epidemiologic studies in endemic areas.
Arias, M; Yeargan, M; Francisco, I; Dangoudoubiyam, S; Becerra, P; Francisco, R; Sánchez-Andrade, R; Paz-Silva, A; Howe, D K
2012-04-30
Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs. Copyright © 2011 Elsevier B.V. All rights reserved.
Jezbera, Jan; Jezberová, Jitka; Kasalický, Vojtěch; Šimek, Karel; Hahn, Martin W.
2013-01-01
Among abundant freshwater Betaproteobacteria, only few groups are considered to be of central ecological importance. One of them is the well-studied genus Limnohabitans and mainly its R-BT subcluster, investigated previously mainly by fluorescence in situ hybridization methods. We designed, based on sequences from a large Limnohabitans culture collection, 18 RLBH (Reverse Line Blot Hybridization) probes specific for different groups within the genus Limnohabitans by targeting diagnostic sequences on their 16 S–23 S rRNA ITS regions. The developed probes covered in sum 92% of the available isolates. This set of probes was applied to environmental DNA originating from 161 different European standing freshwater habitats to reveal the microdiversity (intra-genus) patterns of the Limnohabitans genus along a pH gradient. Investigated habitats differed in various physicochemical parameters, and represented a very broad range of standing freshwater habitats. The Limnohabitans microdiversity, assessed as number of RLBH-defined groups detected, increased significantly along the gradient of rising pH of habitats. 14 out of 18 probes returned detection signals that allowed predictions on the distribution of distinct Limnohabitans groups. Most probe-defined Limnohabitans groups showed preferences for alkaline habitats, one for acidic, and some seemed to lack preferences. Complete niche-separation was indicated for some of the probe-targeted groups. Moreover, bimodal distributions observed for some groups of Limnohabitans, suggested further niche separation between genotypes within the same probe-defined group. Statistical analyses suggested that different environmental parameters such as pH, conductivity, oxygen and altitude influenced the distribution of distinct groups. The results of our study do not support the hypothesis that the wide ecological distribution of Limnohabitans bacteria in standing freshwater habitats results from generalist adaptations of these bacteria
Directory of Open Access Journals (Sweden)
Jan Jezbera
Full Text Available Among abundant freshwater Betaproteobacteria, only few groups are considered to be of central ecological importance. One of them is the well-studied genus Limnohabitans and mainly its R-BT subcluster, investigated previously mainly by fluorescence in situ hybridization methods. We designed, based on sequences from a large Limnohabitans culture collection, 18 RLBH (Reverse Line Blot Hybridization probes specific for different groups within the genus Limnohabitans by targeting diagnostic sequences on their 16 S-23 S rRNA ITS regions. The developed probes covered in sum 92% of the available isolates. This set of probes was applied to environmental DNA originating from 161 different European standing freshwater habitats to reveal the microdiversity (intra-genus patterns of the Limnohabitans genus along a pH gradient. Investigated habitats differed in various physicochemical parameters, and represented a very broad range of standing freshwater habitats. The Limnohabitans microdiversity, assessed as number of RLBH-defined groups detected, increased significantly along the gradient of rising pH of habitats. 14 out of 18 probes returned detection signals that allowed predictions on the distribution of distinct Limnohabitans groups. Most probe-defined Limnohabitans groups showed preferences for alkaline habitats, one for acidic, and some seemed to lack preferences. Complete niche-separation was indicated for some of the probe-targeted groups. Moreover, bimodal distributions observed for some groups of Limnohabitans, suggested further niche separation between genotypes within the same probe-defined group. Statistical analyses suggested that different environmental parameters such as pH, conductivity, oxygen and altitude influenced the distribution of distinct groups. The results of our study do not support the hypothesis that the wide ecological distribution of Limnohabitans bacteria in standing freshwater habitats results from generalist adaptations of
DEFF Research Database (Denmark)
Henriksen, Carsten
2011-01-01
ADHD? Adfærdsvanskelig? Diagnoserne står i kø, når vi i dag skal forklare, hvorfor Emil ikke kan følge med i skolen. Men startskuddet til den videnskabelige opdeling af de børn, der ikke følger flertallets læringskurve, lød allerede for 100 år siden.......ADHD? Adfærdsvanskelig? Diagnoserne står i kø, når vi i dag skal forklare, hvorfor Emil ikke kan følge med i skolen. Men startskuddet til den videnskabelige opdeling af de børn, der ikke følger flertallets læringskurve, lød allerede for 100 år siden....
Gilboa-Garber, Nechama; Lerrer, Batya; Lesman-Movshovich, Efrat; Dgani, Orly
2005-12-01
Human milk, serum, saliva, and seminal fluid glycoproteins (gps) nourish and protect newborn and adult tissues. Their saccharides, which resemble cell membrane components, may block pathogen adhesion and infection. In the present study, they were examined by a battery of lectins from plants, animals, and bacteria, using hemagglutination inhibition and Western blot analyses. The lectins included galactophilic ones from Aplysia gonad, Erythrina corallodendron, Maclura pomifera (MPL), peanut, and Pseudomonas aeruginosa (PA-IL); fucose-binding lectins from Pseudomonas aeruginosa (PA-IIL), Ralstonia solanacearum (RSL), and Ulex europaeus (UEA-I), and mannose/glucose-binding Con A. The results demonstrated the chosen lectin efficiency for differential analysis of human secreted gps as compared to CBB staining. They unveiled the diversity of these body fluid gp glycans (those of the milk and seminal fluid being highest): the milk gps interacted most strongly with PA-IIL, followed by RSL; the saliva gps with RSL, followed by PA-IIL and MPL; the serum gps with Con A and MPL, followed by PA-IIL and RSL, and the seminal plasma gps with RSL and MPL, followed by UEA-I and PA-IIL. The potential usage of these lectins as probes for scientific, industrial, and medical purposes, and for quality control of the desired gps is clearly indicated.
Cogeneration techniques; Les techniques de cogeneration
Energy Technology Data Exchange (ETDEWEB)
NONE
1999-10-01
This dossier about cogeneration techniques comprises 12 parts dealing successively with: the advantages of cogeneration (examples of installations, electrical and thermal efficiency); the combustion turbine (principle, performances, types); the alternative internal combustion engines (principle, types, rotation speed, comparative performances); the different configurations of cogeneration installations based on alternative engines and based on steam turbines (coal, heavy fuel and natural gas-fueled turbines); the environmental constraints of combustion turbines (pollutants, techniques of reduction of pollutant emissions); the environmental constraints of alternative internal combustion engines (gas and diesel engines); cogeneration and energy saving; the techniques of reduction of pollutant emissions (pollutants, unburnt hydrocarbons, primary and secondary (catalytic) techniques, post-combustion); the most-advanced configurations of cogeneration installations for enhanced performances (counter-pressure turbines, massive steam injection cycles, turbo-chargers); comparison between the performances of the different cogeneration techniques; the tri-generation technique (compression and absorption cycles). (J.S.)
Peden, Alexander H.; Ritchie, Diane L.; Head, Mark W.; Ironside, James W.
2006-01-01
Variant Creutzfeldt-Jakob disease (vCJD) differs from other human prion diseases in that the pathogenic prion protein PrPSc can be detected to a greater extent at extraneuronal sites throughout the body, principally within lymphoid tissues. However, a recent study using a high-sensitivity Western blotting technique revealed low levels of PrPSc in skeletal muscle from a quarter of Swiss patients with sporadic CJD (sCJD). This posed the question of whether PrPSc in muscle could also be detected in vCJD, sCJD, and iatrogenic (iCJD) patients from other populations. Therefore, we have used the same high-sensitivity Western blotting technique, in combination with paraffin-embedded tissue blotting, to screen for PrPSc in muscle tissue specimens taken at autopsy from 49 CJD patients in the United Kingdom. These techniques identified muscle PrPSc in 8 of 17 vCJD, 7 of 26 sCJD, and 2 of 5 iCJD patients. Paraffin-embedded tissue blotting analysis showed PrPSc in skeletal muscle in localized anatomical structures that had the morphological and immunohistochemical characteristics of nerve fibers. The detection of PrPSc in muscle tissue from all forms of CJD indicates the possible presence of infectivity in these tissues, suggesting important implications for assessing the potential risk of iatrogenic spread via contaminated surgical instruments. PMID:16507908
Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins.
Ramsey, Kathryn A; Rushton, Zachary L; Ehre, Camille
2016-06-14
Mucins, the heavily-glycosylated proteins lining mucosal surfaces, have evolved as a key component of innate defense by protecting the epithelium against invading pathogens. The main role of these macromolecules is to facilitate particle trapping and clearance while promoting lubrication of the mucosa. During protein synthesis, mucins undergo intense O-glycosylation and multimerization, which dramatically increase the mass and size of these molecules. These post-translational modifications are critical for the viscoelastic properties of mucus. As a result of the complex biochemical and biophysical nature of these molecules, working with mucins provides many challenges that cannot be overcome by conventional protein analysis methods. For instance, their high-molecular-weight prevents electrophoretic migration via regular polyacrylamide gels and their sticky nature causes adhesion to experimental tubing. However, investigating the role of mucins in health (e.g., maintaining mucosal integrity) and disease (e.g., hyperconcentration, mucostasis, cancer) has recently gained interest and mucins are being investigated as a therapeutic target. A better understanding of the production and function of mucin macromolecules may lead to novel pharmaceutical approaches, e.g., inhibitors of mucin granule exocytosis and/or mucolytic agents. Therefore, consistent and reliable protocols to investigate mucin biology are critical for scientific advancement. Here, we describe conventional methods to separate mucin macromolecules by electrophoresis using an agarose gel, transfer protein into nitrocellulose membrane, and detect signal with mucin-specific antibodies as well as infrared fluorescent gel reader. These techniques are widely applicable to determine mucin quantitation, multimerization and to test the effects of pharmacological compounds on mucins.
International Nuclear Information System (INIS)
Okita, Naoyuki; Ashizawa, Daisuke; Ohta, Ryo; Abe, Hideaki; Tanuma, Sei-ichi
2010-01-01
Poly(ADP-ribosyl)ation, which is mainly regulated by poly(ADP-ribose) polymerase (PARP) and poly(ADP-ribose) glycohydrolase (PARG), is a unique protein modification involved in cellular responses such as DNA repair and replication. PARG hydrolyzes glycosidic linkages of poly(ADP-ribose) synthesized by PARP and liberates ADP-ribose residues. Recent studies have suggested that inhibitors of PARG are able to be potent anti-cancer drug. In order to discover the potent and specific Inhibitors of PARG, a quantitative and high-throughput screening assay system is required. However, previous PARG assay systems are not appropriate for high-throughput screening because PARG activity is measured by radioactivities of ADP-ribose residues released from radioisotope (RI)-labeled poly(ADP-ribose). In this study, we developed a non-RI and quantitative assay system for PARG activity based on dot-blot assay using anti-poly(ADP-ribose) and nitrocellulose membrane. By our method, the maximum velocity (V max ) and the michaelis constant (k m ) of PARG reaction were 4.46 μM and 128.33 μmol/min/mg, respectively. Furthermore, the IC50 of adenosine diphosphate (hydroxymethyl) pyrrolidinediol (ADP-HPD), known as a non-competitive PARG inhibitor, was 0.66 μM. These kinetics values were similar to those obtained by traditional PARG assays. By using our assay system, we discovered two novel PARG inhibitors that have xanthene scaffold. Thus, our quantitative and convenient method is useful for a high-throughput screening of PARG specific inhibitors.
Cavazzana, Ilaria; Fredi, Micaela; Ceribelli, Angela; Mordenti, Cristina; Ferrari, Fabio; Carabellese, Nice; Tincani, Angela; Satoh, Minoru; Franceschini, Franco
2016-06-01
To analyze the performance of a line blot assay for the identification of autoantibodies in sera of patients affected by myositis, compared with immunoprecipitation (IP) as gold standard. 66 sera of patients with myositis (23 polymyositis, 8 anti-synthetase syndromes, 29 dermatomyositis and 6 overlap syndromes) were tested by commercial LB (Euroimmun, Lubeck, Germany); 57 sera were analyzed also by IP of K562 cell extract radiolabeled with (35)S-methionine. Inter-rater agreement was calculated with Cohen's k coefficient. Myositis-specific antibodies (MSA) were detected in 36/57 sera (63%) by IP and in 39/66 sera (59%) by LB. The most frequent MSA found by LB were anti-Jo1 and anti-Mi2 found in 15% (10/66) of sera, followed by anti-NXP2 and anti-SRP detected in 106% (7/66) of sera. Anti-TIF1gamma and anti-MDA5 were found in 6 (9%) and 5 sera (7.6%), respectively. A good agreement between methods was found only for anti-TIF1γ, anti-MDA5 and anti-NXP-2 antibodies, while a moderate agreement was estimated for anti-Mi2 and anti-EJ. By contrast, a high discordance rate for the detection of anti-Jo1 antibodies was evident (k: 0.3). Multiple positivity for MSA were found in 11/66 (17%) by LB and 0/57 by IP (p: 0001). Comparing the clinical features of these 11 sera, we found total discrepancies between assays in 3 sera (27.3%), a relative discrepancy due to the occurrence of one discordant autoantibody (not confirmed by IP) in 5 cases (45.5%) and a total discrepancy between LB and IP results, but with a relative concordance with clinical features were found in other 3 sera (27.3%). The semiquantitative results do not support the interpretation of the data. The use of LB assay allowed the detection of new MSA, such as anti-MDA5, anti-MJ and anti-TIF1gamma antibodies, previously not found with routine methods. However, the high prevalence of multiple positivities and the high discondant rate of anti-Jo1 antibodies could create some misinterpretation of the results from the
Directory of Open Access Journals (Sweden)
A Rasul
2013-06-01
Full Text Available Background: The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp. in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.Methods: Reverse line blot (RLB assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.Results: Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.Conclusion: Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001. Risk factor analysis revealed that male (P = 0.03, animals infested by ticks (P = 0.03 and herd composed of sheep only (P = 0.001 were more infected by blood parasites.
Stramer, Susan L; Townsend, Rebecca L; Foster, Gregory A; Johnson, Ramona; Weixlmann, Barbara; Dodd, Roger Y
2018-03-01
Human T-lymphotropic virus (HTLV) blood donation screening has used a dual-testing algorithm beginning with either a chemiluminescent immunoassay or enzyme-linked immunosorbent screening assay (ELISA). Before the availability of a licensed HTLV supplemental assay, repeat-reactive (RR) samples on a first assay (Assay 1) were retested with a second screening assay (Assay 2). Donors with RR results by Assay 2 were deferred from blood donation and further tested using an unlicensed supplemental test to confirm reactivity while nonreactive (NR) donors remained eligible for donation until RR on a subsequent donation. This "dual-test" algorithm was replaced in May 2016 with the requirement that all RRs by Assay 1 be further tested by a licensed HTLV supplemental test (Western blot [WB]). In this study, we have requalified the dual-test algorithm using the available licensed HTLV WB. We tested 100 randomly selected HTLV RRs on screening Assay 1 (Abbott PRISM chemiluminescent immunoassay) but NR on screening Assay 2 (Avioq ELISA) by a Food and Drug Administration-licensed WB (MP Biomedicals) to ensure that no confirmed positives were among those that were RR by Assay 1 but NR by Assay 2. Of the 100 samples evaluated, 79 of 100 were WB seronegative, 21 of 100 indeterminate, and 0 of 100 seropositive. Of the 79 of 100 seronegative specimens, 73 of 79 did not express any bands on WB. We demonstrated that none of the 100 samples RR on Assay 1 but NR on Assay 2 were confirmed positive. This algorithm prevents such donors from requiring further testing and from being deferred. © 2018 AABB.
Navarro, Jose A; Botella, Francisco; Maruhenda, Antonio; Sastre, Pedro; Sánchez-Pina, M Amelia; Pallas, Vicente
2004-05-01
ABSTRACT Nonisotopic molecular dot blot hybridization technique and multiplex reverse transcription-polymerase chain reaction assay for the specific detection of Lettuce big-vein virus (LBVV) and Mirafiori lettuce virus (MiLV) in lettuce tissue were developed. Both procedures were suitable for the specific detection of both viruses in a range of naturally infected lettuce plants from various Spanish production areas and seven different cultivars. The study of the distribution of both viruses in the plant revealed that the highest concentration of LBVV and MiLV occurred in roots and old leaves, respectively. LBVV infection progress in a lettuce production area was faster than that observed for MiLV. In spite of different rates of virus infection progress, most lettuce plants became infected with both viruses about 100 days posttransplant. The appearance of both viruses in lettuce crops was preceded by a peak in the concentration of resting spores and zoosporangia of the fungus vector Olpidium brassicae in lettuce roots.
Directory of Open Access Journals (Sweden)
Santos Terezinha de Jesus Teixeira
2003-01-01
Full Text Available How to handle Western blot (WB seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 constitutes a challenge for blood banks and fam ilies. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA reactive individuals from the hematological center (HEMOCE of Fortaleza (Brazil, examining their serological (WB and molecular (PCR diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22% were positive and 32 (78% were negative for HTLV-1/2. The demographic analysis indicated a trend towards a predominance of males among the seroindeterminate individuals and females in the seropositive ones. The seroindeterminate individuals were younger than the seropositive ones. We did not find any association of these conditions with syphilis, Chagas disease or HIV or hepatitis, and with risk factors such as breast-feeding, blood transfusion, STD (syphilis and IDU.
Directory of Open Access Journals (Sweden)
Terezinha de Jesus Teixeira Santos
Full Text Available How to handle Western blot (WB seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 constitutes a challenge for blood banks and fam ilies. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA reactive individuals from the hematological center (HEMOCE of Fortaleza (Brazil, examining their serological (WB and molecular (PCR diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22% were positive and 32 (78% were negative for HTLV-1/2. The demographic analysis indicated a trend towards a predominance of males among the seroindeterminate individuals and females in the seropositive ones. The seroindeterminate individuals were younger than the seropositive ones. We did not find any association of these conditions with syphilis, Chagas disease or HIV or hepatitis, and with risk factors such as breast-feeding, blood transfusion, STD (syphilis and IDU.
Directory of Open Access Journals (Sweden)
Carlos Eduardo Steiner
2005-09-01
Full Text Available Fragile X syndrome is a frequent genetic disease associated to developmental disorders, including learning disability, mental retardation, behavioral problems and pervasive developmental disorders (autism and related conditions. We studied a sample of 82 individuals (69 males and 13 females presenting with pervasive developmental disorders using three techniques for the diagnosis of fragile X syndrome (FXS. Cytogenetic analysis detected the fragile site in four males, but only one showed a consistent positive rate. Molecular study based on the PCR technique was inconclusive for most females (92.3%, which where latter submitted to Southern blotting analysis, and for one male (1.4%, excluding the FRAXA mutation in the remaining male individuals (98.6%. Molecular tests using the Southern blotting technique confirmed only one positive case (1.2% in a male subject. These results showed that Southern blotting analysis of the FRAXA mutation has the best sensitivity and specificity for the diagnosis of FXS but also validated the PCR technique as a confinable screening test.A síndrome do cromossomo X frágil (SXF é uma doença genética freqüente associada a distúrbios do desenvolvimento neurológico, incluindo dificuldades de aprendizagem, retardo mental, problemas comportamentais e distúrbios invasivos do desenvolvimento (autismo e correlatos. Estudamos uma amostra de 82 indivíduos (69 homens e 13 mulheres apresentando distúrbios invasivos do desenvolvimento, utilizando três técnicas para o diagnóstico da SXF. A análise citogenética detectou a presença do sítio frágil em quatro homens, porém apenas um deles com percentagem consistente. O estudo molecular baseado na técnica da PCR foi inconclusivo para a maioria das mulheres (92,3%, as quais foram posteriormente submetidas a análise por Southern blotting, e para um homem (1,4%, excluindo a mutação FRAXA nos demais homens (98,6%. O teste molecular usando a técnica de Southern
Energy Technology Data Exchange (ETDEWEB)
Wiese, E.
1998-03-13
Most of the dismantling techniques used in a Decontamination and Dismantlement (D and D) project are taken from conventional demolition practices. Some modifications to the techniques are made to limit exposure to the workers or to lessen the spread of contamination to the work area. When working on a D and D project, it is best to keep the dismantling techniques and tools as simple as possible. The workers will be more efficient and safer using techniques that are familiar to them. Prior experience with the technique or use of mock-ups is the best way to keep workers safe and to keep the project on schedule.
International Nuclear Information System (INIS)
Wiese, E.
1998-01-01
Most of the dismantling techniques used in a Decontamination and Dismantlement (D and D) project are taken from conventional demolition practices. Some modifications to the techniques are made to limit exposure to the workers or to lessen the spread of contamination to the work area. When working on a D and D project, it is best to keep the dismantling techniques and tools as simple as possible. The workers will be more efficient and safer using techniques that are familiar to them. Prior experience with the technique or use of mock-ups is the best way to keep workers safe and to keep the project on schedule
Directory of Open Access Journals (Sweden)
P.A. Beck
2007-06-01
Full Text Available Anticorpos monoclonais (AcM para rotavírus bovino foram caracterizados para sua aplicação como ferramenta de diagnóstico, utilizando-se as técnicas de isotipificação, dot-blot, western-blot, imunofluorescência indireta (IFI e ELISA de captura. A caracterização imunoquímica demonstrou que os cinco AcM 1G5, 4F7, 1E12, 4F3 e 3C12 foram do isótipo IgG2a. Pela técnica de dot-blot, os AcM 1G5, 4F7, 1E12, 4F3 detectaram antígenos do rotavírus, em diferentes concentrações, e dois AcM (1E12 e 4F3 reconheceram proteínas virais pela técnica de western-blot. Todos os AcM reagiram positivamente na técnica de IFI em cultivo celular e foram capazes de detectar antígeno viral em amostras fecais bovinas e humanas, pela técnica de ELISA de captura. Identificaram-se dois grupos de AcM, um deles formado pelos AcM 4F7, 1E12 e 1G5, para seu possível uso na detecção de antígeno viral em fezes por meio do ELISA de captura ou dot-blot e outro pelos 4F3 e 3C12, que podem ser usados para detectar antígeno viral em culturas de células por meio de IFI.This work was carried out to characterize and evaluate five bovine rotavirus, monoclonal antibodies (MAbs, as a diagnosis tool, by isotyping, dot-blot, western-blot, indirect immunofluorescence (IFI and ELISA techniques. The immunochemistry characterization showed that all five MAbs (4F7, 4F3, 1G5, 1E12 and 3C12 were IgG2a isotype. The dot-blot immunoassay showed that 1G5, 4F7, 1E12 and 4F3 detected viral antigen in different concentrations and two MAbs (1E12 and 4F3 recognized viral proteins by western-blot. All MAbs detected viral antigen in bovine and human fecal samples by capture ELISA technique and viral antigen in infected MA-104 cell culture by IFI. In conclusion, two groups of Mabs were indetified: one with Mabs 4F7, 1E12 and 1G5 showed the best results to detect rotavirus antigen in fecal samples by capture ELISA or dot-blot techniques assay and other with 4F3 and 3C12 which may be used to
Tzeng, C-C; Tsai, L-P; Chang, Y-K; Hung, Y-J; Chang, Y-Y; Su, Y-P; Jiang, J-J; Liang, H-M
2017-08-01
Here, we review the results of Southern blotting analyses of the FMR1 gene performed in our reference laboratory in Taiwan over a 15-year period. In total, 725 high-risk women with a family history of fragile X syndrome (FXS) or idiopathic intellectual disability, 3911 low-risk pregnant women without such family history, and prenatal diagnosis data for 32 foetuses from 24 carrier mothers were included. Only 2 carriers were in the low-risk group, which indicated a prevalence of 1 of 1955 women (95% confidence interval: 1/7156-1/539). A total of 100 carriers were found to be in the high-risk group, thus revealing a significantly higher frequency than the low-risk group (100/725 vs 2/3911, P<0.0001). Eight of the 14 foetuses that inherited the maternal mutant allele were verified to have a full mutation, with the smallest maternal pre-mutation allele carrying 56 CGG repeats. The overall findings confirmed that the carrier prevalence among low-risk women in Taiwan is significantly lower than that reported in western countries. Therefore, the most important step for preventing FXS in Taiwan would be to focus on high-risk women by promoting general awareness of this disease and spreading knowledge regarding the benefits of carrier screening and prenatal testing. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Bracco, Gianangelo
2013-01-01
The book describes the experimental techniques employed to study surfaces and interfaces. The emphasis is on the experimental method. Therefore all chapters start with an introduction of the scientific problem, the theory necessary to understand how the technique works and how to understand the results. Descriptions of real experimental setups, experimental results at different systems are given to show both the strength and the limits of the technique. In a final part the new developments and possible extensions of the techniques are presented. The included techniques provide microscopic as well as macroscopic information. They cover most of the techniques used in surface science.
Chao, T.T.; Sanzolone, R.F.
1992-01-01
Sample decomposition is a fundamental and integral step in the procedure of geochemical analysis. It is often the limiting factor to sample throughput, especially with the recent application of the fast and modern multi-element measurement instrumentation. The complexity of geological materials makes it necessary to choose the sample decomposition technique that is compatible with the specific objective of the analysis. When selecting a decomposition technique, consideration should be given to the chemical and mineralogical characteristics of the sample, elements to be determined, precision and accuracy requirements, sample throughput, technical capability of personnel, and time constraints. This paper addresses these concerns and discusses the attributes and limitations of many techniques of sample decomposition along with examples of their application to geochemical analysis. The chemical properties of reagents as to their function as decomposition agents are also reviewed. The section on acid dissolution techniques addresses the various inorganic acids that are used individually or in combination in both open and closed systems. Fluxes used in sample fusion are discussed. The promising microwave-oven technology and the emerging field of automation are also examined. A section on applications highlights the use of decomposition techniques for the determination of Au, platinum group elements (PGEs), Hg, U, hydride-forming elements, rare earth elements (REEs), and multi-elements in geological materials. Partial dissolution techniques used for geochemical exploration which have been treated in detail elsewhere are not discussed here; nor are fire-assaying for noble metals and decomposition techniques for X-ray fluorescence or nuclear methods be discussed. ?? 1992.
Laboratoriediagnostik af infektion forårsaget af Borrelia burgdorferi
DEFF Research Database (Denmark)
Dessau, Ram; Bangsborg, Jette Marie; Jensen, Tove P Ejlertsen
2006-01-01
The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot as confi......The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot...... as confirmatory testing is not recommended since the contribution to the diagnostic specificity is only marginal. Predictive values of Lyme serology are presented, based on the estimated prevalence of the different stages of Lyme disease in Denmark....
Marcia Pinheiro
2015-01-01
In this paper, we discuss three translation techniques: literal, cultural, and artistic. Literal translation is a well-known technique, which means that it is quite easy to find sources on the topic. Cultural and artistic translation may be new terms. Whilst cultural translation focuses on matching contexts, artistic translation focuses on matching reactions. Because literal translation matches only words, it is not hard to find situations in which we should not use this technique. Because a...
Evaluación de la técnica Western blot para la detección de antígenos de Hymenolepis nana
Directory of Open Access Journals (Sweden)
Flora Chávez-Salas
2013-04-01
Full Text Available Este trabajo tuvo como objetivo evaluar la técnica de inmunoelectrotransferencia (Western Blot para detectar los antígenos específicos de excreción/secreción de Hymenolepis nana en sueros de pacientes con himenolepiosis y con otras helmintiosis confirmadas. Se utilizó a Mesocricetus auratus “hamster” para obtener ejemplares adultos de H. nana. Los antígenos de excreción/secreción fueron obtenidos en el medio MEM (Minimum Essential Medium Eagle, y enfrentados con un grupo de sueros de pacientes con himenolepiosis confirmada para evaluar su calidad inmunológica y con sueros individuales de pacientes con himenolepiosis y con otras helmintiosis confirmadas para detectar mediante la técnica de “Western Blot”, los antígenos específicos de este cestode. El grupo de sueros de pacientes con himenolepiosis confirmada reconoció las bandas antigénicas de 50,1; 42,6; 38,9; 32,9; 26,3; 22,4 y 18,6 kDa; sin embargo, los sueros individuales reconocieron diferente número de bandas, siendo la de 50,1 KDa la que fue reconocida por todos ellos. Los sueros de pacientes con helmintiosis confirmadas no reconocieron la banda de 50,1 kDa; sin embargo, dieron reacción cruzada con algunas de las demás bandas, a excepción de los sueros de pacientes con cisticercosis que no reconocieron a ninguna de las bandas de estos antígenos. Se concluye que el antígeno de excreción/secreción de H. nana de 50,1 kDa es específico de este cestode por ser reconocido por todos los sueros de pacientes con himenolepiosis confirmada y no por sueros de pacientes con otras helmintiosis utilizando la técnica de “Western Blot”.
Specific radiography technique
International Nuclear Information System (INIS)
Abdul Nassir Ibrahim; Azali Muhammad; Ab. Razak Hamzah; Abd. Aziz Mohamed; Mohamad Pauzi Ismail
2008-01-01
Beside radiography testing using x-ray machine and gamma source, there are several technique that developed specifically to complete the testing that cannot be done with the two earlier. This technique was specific based on several factor, for the example, the advantages of neutron and electron using to show the image was unique compare to x-ray and gamma. Besides that, these special radiography techniques maybe differ in how to detect the radiation get through the object. These technique can used to inspect thin or specimen that contained radioactive material. There are several technique will discussed in this chapter such as neutron radiography, electron radiography, fluoroscopy and also autoradiography.
A cost-effective device for the rapid transfer of gel-separated proteins onto membranes.
Tam, Hann W; Huang, Yu-Chen; Tam, Ming F
2009-03-01
We describe here the fabrication of a cost-effective semi-dry blotting apparatus for the transfer of proteins onto membranes. Graphite sheets were used as electrodes. Protein mixtures were separated on NuPAGE 4% to 12% polyacrylamide gradient gels. With a Tris-bicine buffer, we demonstrated that close to 80% of the proteins with apparent molecular mass of 80kDa or less were removed from the gels after 8min of blotting. The process is much faster than the techniques reported previously in the literature.
African Journals Online (AJOL)
.
Minimally invasive techniques are becoming popular choice for the recent times. These techniques are lowering the cost and giving the best cosmetic results. For cardiovascular surgery these techniques are much more newer and much more unknown. Open lumbar sympathectomy for certain indications is a very well ...
Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo
2017-01-01
An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic ® cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex ® resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent. Copyright © 2016 Elsevier GmbH. All rights reserved.
Buchman, Vladimir L; Cooper-Knock, Johnathan; Connor-Robson, Natalie; Higginbottom, Adrian; Kirby, Janine; Razinskaya, Olga D; Ninkina, Natalia; Shaw, Pamela J
2013-04-08
Sizing of GGGGCC hexanucleotide repeat expansions within the C9ORF72 locus, which account for approximately 10% of all amyotrophic lateral sclerosis (ALS) cases, is urgently required to answer fundamental questions about mechanisms of pathogenesis in this important genetic variant. Currently employed PCR protocols are limited to discrimination between the presence and absence of a modified allele with more than 30 copies of the repeat, while Southern hybridisation-based methods are confounded by the somatic heterogeneity commonly present in blood samples, which might cause false-negative or ambiguous results. We describe an optimised Southern hybridisation-based protocol that allows confident detection of the presence of a C9ORF72 repeat expansion alongside independent assessment of its heterogeneity and the number of repeat units. The protocol can be used with either a radiolabeled or non-radiolabeled probe. Using this method we have successfully sized the C9ORF72 repeat expansion in lymphoblastoid cells, peripheral blood, and post-mortem central nervous system (CNS) tissue from ALS patients. It was also possible to confidently demonstrate the presence of repeat expansion, although of different magnitude, in both C9ORF72 alleles of the genome of one patient. The suggested protocol has sufficient advantages to warrant adoption as a standard for Southern blot hybridisation analysis of GGGGCC repeat expansions in the C9ORF72 locus.
Uranium exploration techniques
International Nuclear Information System (INIS)
Nichols, C.E.
1984-01-01
The subject is discussed under the headings: introduction (genetic description of some uranium deposits; typical concentrations of uranium in the natural environment); sedimentary host rocks (sandstones; tabular deposits; roll-front deposits; black shales); metamorphic host rocks (exploration techniques); geologic techniques (alteration features in sandstones; favourable features in metamorphic rocks); geophysical techniques (radiometric surveys; surface vehicle methods; airborne methods; input surveys); geochemical techniques (hydrogeochemistry; petrogeochemistry; stream sediment geochemistry; pedogeochemistry; emanometry; biogeochemistry); geochemical model for roll-front deposits; geologic model for vein-like deposits. (U.K.)
Directory of Open Access Journals (Sweden)
Manouchehr Moshtagh Khorasani
2012-07-01
Full Text Available There are numerous manuscripts, poems and stories that describe, specifically and in detail, the different techniques used in Persian swordsmanship. The present article explains the origins and the techniques of Persian swordsmanship. The article also describes the traditional code of conduct for Persian warriors. Additionally, it describes an array of techniques that were deployed in actual combat in Iran’s history. Some of these techniques are represented via the miniatures that are reproduced herein. This is the first article on Persian swordsmanship published in any periodical.
Radiation scattering techniques
International Nuclear Information System (INIS)
Edmonds, E.A.
1986-01-01
Radiation backscattering techniques are useful when access to an item to be inspected is restricted to one side. These techniques are very sensitive to geometrical effects. Scattering processes and their application to the determination of voids, thickness measuring, well-logging and the use of x-ray fluorescence techniques are discussed. (U.K.)
Laboratoriediagnostik af infektion forårsaget af Borrelia burgdorferi
DEFF Research Database (Denmark)
Dessau, Ram B; Bangsborg, Jette M; Jensen, Tove P Ejlertsen
2006-01-01
The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot as confi......The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot...... as confirmatory testing is not recommended since the contribution to the diagnostic specificity is only marginal. Predictive values of Lyme serology are presented, based on the estimated prevalence of the different stages of Lyme disease in Denmark. Udgivelsesdato: 2006-Aug-21...
Preference for different relaxation techniques by COPD patients: comparison between six techniques
Directory of Open Access Journals (Sweden)
Hyl
2016-09-01
Full Text Available Michael E Hyland,1 David MG Halpin,2 Sue Blake,3 Clare Seamark,3 Margaret Pinnuck,3 David Ward,3 Ben Whalley,1 Colin J Greaves,4 Adam L Hawkins,5 Dave Seamark3 1School of Psychology, University of Plymouth, Plymouth, 2Department of Respiratory Medicine, Royal Devon and Exeter Hospital, Exeter, 3Honiton Group Practice, Honiton, 4University of Exeter Medical School, Exeter, 5GSK House, Brentford, UK Background: A review of the effectiveness of relaxation techniques for chronic obstructive pulmonary disease patients has shown inconsistent results, but studies have varied in terms of technique and outcome measures. Aim: To determine patient preference for different relaxation techniques. Methods: Chronic obstructive pulmonary disease patients were presented with six techniques via a DVD and asked to rate the techniques in terms of effectiveness, rank in order of likely use, and comment. Results: Patients differed in the technique preferred and reason for that preference, but the most commonly preferred technique both for effectiveness and ease of use was “thinking of a nice place” followed by progressive relaxation and counting. Familiarity and ease of activity were commonly given reasons for preference. Conclusion: Rather than providing patients with a single technique that they might find difficult to implement, these results suggest that it would be better to give a choice. “Thinking of a nice place” is a popular but under-investigated technique. Keywords: COPD exacerbation, anxiety, relaxation techniques
Training techniques for industry
International Nuclear Information System (INIS)
Jones, D.W.
1978-01-01
The advantages and disadvantages of commonly used training techniques in relation to cost-effective, prevention-oriented Quality Assurance are examined. Important questions are whether training techniques teach cost effectiveness and whether the techniques are, themselves, cost effective. To answer these questions, criteria for evaluating teaching techniques for cost effectiveness were developd, and then commonly used techniques are evaluated in terms of specific training program objectives. Motivation of personnel is also considered important to the success of a training program, and methods are outlined by which recognition of the academic quality of industrial training can be used as a motivational technique
Directory of Open Access Journals (Sweden)
C Soldani
2009-06-01
Full Text Available Apoptosis occurring in atherosclerotic lesions has been suggested to be involved in the evolution and the structural stability of the plaques. It is still a matter of debate whether apoptosis mainly involves vascular smooth muscle cells (vSMCs in the fibrous tissue or inflammatory (namely foam cells, thus preferentially affecting the cell-poor lipid core of the atherosclerotic plaques. The aim of the present investigation was to detect the presence of apoptotic cells and to estimate their percentage in a series of atherosclerotic plaques obtained either by autopsy or during surgical atherectomy. Apoptotic cells were identified on paraffinembedded sections on the basis of cell nuclear morphology after DNA staining and/or by cytochemical reactions (TUNEL assay, immunodetection of the proteolytic poly (ADP-ribose polymerase-1 [PARP-1] fragment; biochemical procedures (identifying DNA fragmentation or PARP-1 proteolysis were also used. Indirect immunofluorescence techniques were performed to label specific antigens for either vSMCs or macrophages (i.e., the cells which are most likely prone to apoptosis in atherosclerotic lesions: the proper selection of fluorochrome labeling allowed the simultaneous detection of the cell phenotype and the apoptotic characteristics, by multicolor fluorescence techniques. Apoptotic cells proved to be less than 5% of the whole cell population, in atherosclerotic plaque sections: this is, in fact, a too low cell fraction to be detected by widely used biochemical methods, such as agarose gel electrophoresis of low-molecular-weight DNA or Western-blot analysis of PARP-1 degradation. Most apoptotic cells were of macrophage origin, and clustered in the tunica media, near or within the lipid-rich core; only a few TUNEL-positive cells were labeled for antigens specific for vSMCs. These results confirm that, among the cell populations in atherosclerotic plaques, macrophage foam-cells are preferentially involved in apoptosis
Automatic DNA Diagnosis for 1D Gel Electrophoresis Images using Bio-image Processing Technique.
Intarapanich, Apichart; Kaewkamnerd, Saowaluck; Shaw, Philip J; Ukosakit, Kittipat; Tragoonrung, Somvong; Tongsima, Sissades
2015-01-01
DNA gel electrophoresis is a molecular biology technique for separating different sizes of DNA fragments. Applications of DNA gel electrophoresis include DNA fingerprinting (genetic diagnosis), size estimation of DNA, and DNA separation for Southern blotting. Accurate interpretation of DNA banding patterns from electrophoretic images can be laborious and error prone when a large number of bands are interrogated manually. Although many bio-imaging techniques have been proposed, none of them can fully automate the typing of DNA owing to the complexities of migration patterns typically obtained. We developed an image-processing tool that automatically calls genotypes from DNA gel electrophoresis images. The image processing workflow comprises three main steps: 1) lane segmentation, 2) extraction of DNA bands and 3) band genotyping classification. The tool was originally intended to facilitate large-scale genotyping analysis of sugarcane cultivars. We tested the proposed tool on 10 gel images (433 cultivars) obtained from polyacrylamide gel electrophoresis (PAGE) of PCR amplicons for detecting intron length polymorphisms (ILP) on one locus of the sugarcanes. These gel images demonstrated many challenges in automated lane/band segmentation in image processing including lane distortion, band deformity, high degree of noise in the background, and bands that are very close together (doublets). Using the proposed bio-imaging workflow, lanes and DNA bands contained within are properly segmented, even for adjacent bands with aberrant migration that cannot be separated by conventional techniques. The software, called GELect, automatically performs genotype calling on each lane by comparing with an all-banding reference, which was created by clustering the existing bands into the non-redundant set of reference bands. The automated genotype calling results were verified by independent manual typing by molecular biologists. This work presents an automated genotyping tool from DNA
Automatic DNA Diagnosis for 1D Gel Electrophoresis Images using Bio-image Processing Technique
2015-01-01
Background DNA gel electrophoresis is a molecular biology technique for separating different sizes of DNA fragments. Applications of DNA gel electrophoresis include DNA fingerprinting (genetic diagnosis), size estimation of DNA, and DNA separation for Southern blotting. Accurate interpretation of DNA banding patterns from electrophoretic images can be laborious and error prone when a large number of bands are interrogated manually. Although many bio-imaging techniques have been proposed, none of them can fully automate the typing of DNA owing to the complexities of migration patterns typically obtained. Results We developed an image-processing tool that automatically calls genotypes from DNA gel electrophoresis images. The image processing workflow comprises three main steps: 1) lane segmentation, 2) extraction of DNA bands and 3) band genotyping classification. The tool was originally intended to facilitate large-scale genotyping analysis of sugarcane cultivars. We tested the proposed tool on 10 gel images (433 cultivars) obtained from polyacrylamide gel electrophoresis (PAGE) of PCR amplicons for detecting intron length polymorphisms (ILP) on one locus of the sugarcanes. These gel images demonstrated many challenges in automated lane/band segmentation in image processing including lane distortion, band deformity, high degree of noise in the background, and bands that are very close together (doublets). Using the proposed bio-imaging workflow, lanes and DNA bands contained within are properly segmented, even for adjacent bands with aberrant migration that cannot be separated by conventional techniques. The software, called GELect, automatically performs genotype calling on each lane by comparing with an all-banding reference, which was created by clustering the existing bands into the non-redundant set of reference bands. The automated genotype calling results were verified by independent manual typing by molecular biologists. Conclusions This work presents an
Growth factors regulate glutamine synthetase activity in ...
African Journals Online (AJOL)
Khaled
2012-07-10
Jul 10, 2012 ... glutamate and ammonia, which in turn, cells are supplied with ammonia ... out to determine the maximum growth time at which cells will be .... Western blot technique for detection the glutamine synthetase enzyme. Lane 1;.
Directory of Open Access Journals (Sweden)
Tetsuya Okuda
2017-02-01
Full Text Available Protein modification by O-linked N-acetylglucosamine (O-GlcNAcylation is one of the post transcriptional modifications occurring on cellular proteins. This paper provides a data set relating to the O-GlcNAcylation of cellular proteins detected by RL2 and CTD110.6 antibodies, which are commonly used for detection of protein O-GlcNAcylation, in 2-deoxy-d-glucose (2DG-treated human teratocarcinoma NCCIT cells in support of the research article entitled “A novel, promoter-based, target-specific assay identifies 2-deoxy-d-glucose as an inhibitor of globotriaosylceramide biosynthesis” (Okuda et al., 2009 [1]. The main article described a suppressive effect of 2DG on an Sp1 target gene in NCCIT cells and discussed the relationship between the effect of 2DG and O-GlcNAcylation status of Sp1. The data in this paper complements this relationship by Western blotting and clearly showed that the 2DG treatment increased O-GlcNAcylation of cellular proteins in NCCIT cells, whereas the RL2 and CTD110.6 epitopes were detected in a different manner. The RL2 epitope was detected on Sp1 during 2DG treatment, and the level was transiently increased at 24 h. In contrast, the CTD110.6 epitope became detectable on Sp1 over 72 h after 2DG treatment, and then the other proteins containing CTD110.6 epitopes also appeared in the cell lysates and the anti-Sp1 antibody precipitates.
Pulse holographic measurement techniques
International Nuclear Information System (INIS)
Kim, Cheol Jung; Baik, Seong Hoon; Hong, Seok Kyung; Kim, Jeong Moog; Kim, Duk Hyun
1992-01-01
With the development of laser, remote inspection techniques using laser have been growing on. The inspection and measurement techniques by pulse holography are well-established technique for precise measurement, and widely used in various fields of industry now. In nuclear industry, this technology is practically used because holographic inspection is remote, noncontact, and precise measurement technique. In relation to remote inspection technology in nuclear industry, state-of-the art of pulse HNDT (Holographic non-destructive testing) and holographic measurement techniques are examined. First of all, the fundamental principles as well as practical problems for applications are briefly described. The fields of pulse holography have been divided into the HNDT, flow visualization and distribution study, and other application techniques. Additionally holographic particle study, bubble chamber holography, and applications to other visualization techniques are described. Lastly, the current status for the researches and applications of pulse holography to nuclear industry which are carried out actively in Europe and USA, is described. (Author)
SEM-based characterization techniques
International Nuclear Information System (INIS)
Russell, P.E.
1986-01-01
The scanning electron microscope is now a common instrument in materials characterization laboratories. The basic role of the SEM as a topographic imaging system has steadily been expanding to include a variety of SEM-based analytical techniques. These techniques cover the range of basic semiconductor materials characterization to live-time device characterization of operating LSI or VLSI devices. This paper introduces many of the more commonly used techniques, describes the modifications or additions to a conventional SEM required to utilize the techniques, and gives examples of the use of such techniques. First, the types of signals available from a sample being irradiated by an electron beam are reviewed. Then, where applicable, the type of spectroscopy or microscopy which has evolved to utilize the various signal types are described. This is followed by specific examples of the use of such techniques to solve problems related to semiconductor technology. Techniques emphasized include: x-ray fluorescence spectroscopy, electron beam induced current (EBIC), stroboscopic voltage analysis, cathodoluminescnece and electron beam IC metrology. Current and future trends of some of the these techniques, as related to the semiconductor industry are discussed
DNA polymorphism in the living fossil Ginkgo biloba from the eastern United States.
Kuddus, Ruhul H; Kuddus, Nayema N; Dvorchik, Igor
2002-02-01
Random amplified polymorphic DNA (RAPD) analysis is a valuable tool in studying inter- and intra-specific genetic variations, patterns of gene expression, and for the identification of specific genes using nearly isogenic variants. Here we used RAPD analysis to study the genetic variation in Ginkgo biloba grown in the eastern United States. Our results support the evidence that Southern blot hybridization of RAPD using probes made from cloned DNA fragments allows a more accurate analysis of the RAPD pattern than dye-stained gels or Southern blot hybridization of RAPD blots using probes made from purified PCR products. Using these techniques, we observed a high degree of relatedness among plants grown in certain localities although significant genetic variation may exist in the species, and could be a possible explanation for the observed variations in the efficacy of medications derived from G. biloba extract.
Nuclear techniques in hydrology
International Nuclear Information System (INIS)
Moser, H.
1976-01-01
The nuclear techniques used in hydrology are usually tracer techniques based on the use of nuclides either intentionally introduced into, or naturally present in the water. The low concentrations of these nuclides, which must be detected in groundwater and surface water, require special measurement techniques for the concentrations of radioactive or of stable nuclides. The nuclear techniques can be used most fruitfully in conjunction with conventional methods for the solution of problems in the areas of hydrology, hydrogeology and glacier hydrology. Nuclear techniques are used in practice in the areas of prospecting for water, environment protection and engineering hydrogeology. (orig.) [de
Simon, Stephane; Lumley, Philip; Tomson, Phillip; Pertot, Wilhelm-Joseph; Machtou, Pierre
2008-03-01
Crown down preparation is the most known and described technique since the introduction of Nickel Titanium (NiTi) rotary instruments in endodontics. This technique gives good results but has limitations, such as not addressing the initial anatomy of oval or dumb-bell shaped canals. The specific design of the Protaper instruments allows use of them with a different technique and, specifically, with a brushing motion in the body of the canal. The recent introduction of hand Protaper files has expanded the range of application of this system, especially in curved canals. The 'hybrid technique', using rotary and hand files, and the advantages of the combination of both instruments, are clearly described in this article. Used with this technique, the Protaper is a very safe system to use, and more controllable, for both inexperienced and experienced practitioners alike, than other systems. To understand the precautions needed with rotary files, and how to use them to preserve the anatomy of the canal and get a tapered shaping, even in severely curved canals.
Energy Technology Data Exchange (ETDEWEB)
Saint, A.; Laird, J.S.; Bardos, R.A.; Legge, G.J.F. [Melbourne Univ., Parkville, VIC (Australia). School of Physics; Nishijima, T.; Sekiguchi, H. [Electrotechnical Laboratory, Tsukuba (Japan).
1993-12-31
Since the development of Scanning Transmission Microscopy (STIM) imaging in 1983 many low current beam techniques have been developed for the scanning (ion) microprobe. These include STIM tomography, Ion Beam Induced Current, Ion Beam Micromachining and Microlithography and Ionoluminense. Most of these techniques utilise beam currents of 10{sup -15} A down to single ions controlled by beam switching techniques This paper will discuss some of the low beam current techniques mentioned above, and indicate, some of their recent applications at MARC. A new STIM technique will be introduced that can be used to obtain Z-contrast with STIM resolution. 4 refs., 3 figs.
Energy Technology Data Exchange (ETDEWEB)
Saint, A; Laird, J S; Bardos, R A; Legge, G J.F. [Melbourne Univ., Parkville, VIC (Australia). School of Physics; Nishijima, T; Sekiguchi, H [Electrotechnical Laboratory, Tsukuba (Japan).
1994-12-31
Since the development of Scanning Transmission Microscopy (STIM) imaging in 1983 many low current beam techniques have been developed for the scanning (ion) microprobe. These include STIM tomography, Ion Beam Induced Current, Ion Beam Micromachining and Microlithography and Ionoluminense. Most of these techniques utilise beam currents of 10{sup -15} A down to single ions controlled by beam switching techniques This paper will discuss some of the low beam current techniques mentioned above, and indicate, some of their recent applications at MARC. A new STIM technique will be introduced that can be used to obtain Z-contrast with STIM resolution. 4 refs., 3 figs.
Wieczorek, Piotr; Ligor, Magdalena; Buszewski, Bogusław
Electromigration techniques, including capillary electrophoresis (CE), are widely used for separation and identification of compounds present in food products. These techniques may also be considered as alternate and complementary with respect to commonly used analytical techniques, such as high-performance liquid chromatography (HPLC), or gas chromatography (GC). Applications of CE concern the determination of high-molecular compounds, like polyphenols, including flavonoids, pigments, vitamins, food additives (preservatives, antioxidants, sweeteners, artificial pigments) are presented. Also, the method developed for the determination of proteins and peptides composed of amino acids, which are basic components of food products, are studied. Other substances such as carbohydrates, nucleic acids, biogenic amines, natural toxins, and other contaminations including pesticides and antibiotics are discussed. The possibility of CE application in food control laboratories, where analysis of the composition of food and food products are conducted, is of great importance. CE technique may be used during the control of technological processes in the food industry and for the identification of numerous compounds present in food. Due to the numerous advantages of the CE technique it is successfully used in routine food analysis.
Vanasse, George A
2013-01-01
Spectrometric Techniques, Volume IV discusses three widely diversified areas of spectrometric techniques. The book focuses on three spectrometric methods. Chapter 1 discusses the phenomenology and applications of Coherent Anti-Stokes Raman Spectroscopy (CARS), the most commonly used optical technique that exploit the Raman effect. The second chapter is concerned with diffraction gratings and mountings for the Vacuum Ultraviolet Spectral Region. Chapter 3 accounts the uses of mass spectrometry, detectors, types of spectrometers, and ion sources. Physicists and chemists will find the book a go
Vanasse, George A
2013-01-01
Spectrometric Techniques, Volume II provides information pertinent to vacuum ultraviolet techniques to complete the demonstration of the diversity of methods available to the spectroscopist interested in the ultraviolet visible and infrared spectral regions. This book discusses the specific aspects of the technique of Fourier transform spectroscopy.Organized into five chapters, this volume begins with an overview of the large number of systematic effects in the recording of an interferogram. This text then examines the design approach for a Fourier transform spectrometer with focus on optics.
Vanasse, George A
2013-01-01
Spectrometric Techniques, Volume III presents the applications of spectrometric techniques to atmospheric and space studies. This book reviews the spectral data processing and analysis techniques that are of broad applicability.Organized into five chapters, this volume begins with an overview of the instrumentation used for obtaining field data. This text then reviews the contribution that space-borne spectroscopy in the thermal IR has made to the understanding of the planets. Other chapters consider the instruments that have recorded the planetary emission spectra. This book discusses as well
Quality Attribute Techniques Framework
Chiam, Yin Kia; Zhu, Liming; Staples, Mark
The quality of software is achieved during its development. Development teams use various techniques to investigate, evaluate and control potential quality problems in their systems. These “Quality Attribute Techniques” target specific product qualities such as safety or security. This paper proposes a framework to capture important characteristics of these techniques. The framework is intended to support process tailoring, by facilitating the selection of techniques for inclusion into process models that target specific product qualities. We use risk management as a theory to accommodate techniques for many product qualities and lifecycle phases. Safety techniques have motivated the framework, and safety and performance techniques have been used to evaluate the framework. The evaluation demonstrates the ability of quality risk management to cover the development lifecycle and to accommodate two different product qualities. We identify advantages and limitations of the framework, and discuss future research on the framework.
DEFF Research Database (Denmark)
Csati, Anett; Tajti, Janos; Tuka, Bernadett
2012-01-01
, Western blot technique was used to demonstrate the existence of CGRP receptor components in rat SPG. CGRP immunoreactive fibers were frequently found intraganglionic in the SPG in the vicinity of neurons. CLR immunoreactivity was observed in satellite glial cells (SGCs) as well as in nerve fibers......, but not in neurons. RAMP1 immunoreactivity was localized in many neurons and SGCs. Thus, the two CGRP receptor components together were found in the SGCs. In addition, Western blot revealed the presence of RAMP1 and CLR in rat SPG. Our results suggest a possible sensory influence in the parasympathetic cranial...
Altvorst, van A.C.; Koehorst, H.; Jong, de J.; Dons, M.M.
1996-01-01
Transgenic carnation plants were obtained after infection of petal explants with the supervirulent Agrobacterium tumefaciens strain AGLO. Southern blot techniques confirmed the transgenic nature of four transformed plants. The expression of the gus gene was verified in these plants by histochemical
Zhang, Lahong; Dai, Yibei; Chen, Jiahuan; Hong, Liquan; Liu, Yuhua; Ke, Qiang; Chen, Yiwen; Cai, Chengsong; Liu, Xia; Chen, Zhaojun
2018-01-01
A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. © 2017 Wiley Periodicals, Inc.
Directory of Open Access Journals (Sweden)
Camila Cánepa
2015-10-01
Full Text Available Background: indeterminate Western blot (WB patterns are a major concern for diagnosis of human T-cell lymphotropic virus type 1 (HTLV-1 infection, even in non-endemic areas. Objectives: (a to define the prevalence of indeterminate WB among different populations from Argentina; (b to evaluate if low proviral load (PVL is associated with indeterminate WB profiles; and (c to describe mutations in LTR and tax sequence of these cases. Results: Among 2031 samples, 294 were reactive by screening. Of them, 48 (16.3% were WB indeterminate and of those 15 (31.3% were PCR+. Quantitative real-time PCR (qPCR was performed to 52 HTLV-1+ samples, classified as Group 1 (G1: 25 WB+ samples from individuals with pathologies; Group 2 (G2: 18 WB+ samples from asymptomatic carriers (AC; and Group 3 (G3: 9 seroindeterminate samples from AC. Median PVL was 4.78, 2.38, and 0.15 HTLV-1 copies/100 PBMCs, respectively; a significant difference (p=0.003 was observed. Age and sex were associated with PVL in G1 and G2, respectively. Mutations in the distal and central regions of Tax Responsive Elements (TRE 1 and 2 of G3 were observed, though not associated with PVL.The 8403A>G mutation of the distal region, previously related to high PVL, was absent in G3 but present in 50% of WB+ samples (p = 0.03. Conclusions: indeterminateWBresults confirmed later as HTLV-1 positive may be associated with low PVL levels. Mutations in LTR and tax are described; their functional relevance remains to be determined.
Directory of Open Access Journals (Sweden)
Mohamed Wishahi
2015-08-01
Full Text Available ABSTRACTBackground:Uretero-ileal anastomotic stricture (UIAS is a urological complication after ileal neobladder, the initial management being endourological intervention. If this fails or stricture recurs, surgical intervention will be indicated.Design and Participants:From 1994 to 2013, 129 patients were treated for UIAS after unsuccessful endourological intervention. Unilateral UIAS was present in 101 patients, and bilateral in 28 patients; total procedures were 157. The previous ileal neobladder techniques were Hautmann neobladder, detubularized U shape, or spherical shape neobladder.Surgical procedures:Dipping technique was performed in 74 UIAS. Detour technique was done in 60 renal units. Ileal Bladder flap was indicated in 23 renal units. Each procedure ended with insertion of double J, abdominal drain, and indwelling catheter.Results:Follow-up was done for 12 to 36 months. Patency of the anastomosis was found in 91.7 % of cases. Thirteen patients (8.3% underwent antegrade dilatation and insertion of double J.Conclusion:After endourological treatment for uretero-ileal anastomotic failure, basically three techniques may be indicated: dipping technique, detour technique, and ileal bladder flap. The indications are dependent on the length of the stenotic/dilated ureteral segment. Better results for long length of stenotic ureter are obtained with detour technique; for short length stenotic ureter dipping technique; when the stenotic segment is 5 cm or more with a short ureter, the ileal tube flap is indicated. The use of double J stent is mandatory in the majority of cases. Early intervention is the rule for protecting renal units from progressive loss of function.
Emerging optical nanoscopy techniques
Directory of Open Access Journals (Sweden)
Montgomery PC
2015-09-01
Full Text Available Paul C Montgomery, Audrey Leong-Hoi Laboratoire des Sciences de l'Ingénieur, de l'Informatique et de l'Imagerie (ICube, Unistra-CNRS, Strasbourg, France Abstract: To face the challenges of modern health care, new imaging techniques with subcellular resolution or detection over wide fields are required. Far field optical nanoscopy presents many new solutions, providing high resolution or detection at high speed. We present a new classification scheme to help appreciate the growing number of optical nanoscopy techniques. We underline an important distinction between superresolution techniques that provide improved resolving power and nanodetection techniques for characterizing unresolved nanostructures. Some of the emerging techniques within these two categories are highlighted with applications in biophysics and medicine. Recent techniques employing wider angle imaging by digital holography and scattering lens microscopy allow superresolution to be achieved for subcellular and even in vivo, imaging without labeling. Nanodetection techniques are divided into four subcategories using contrast, phase, deconvolution, and nanomarkers. Contrast enhancement is illustrated by means of a polarized light-based technique and with strobed phase-contrast microscopy to reveal nanostructures. Very high sensitivity phase measurement using interference microscopy is shown to provide nanometric surface roughness measurement or to reveal internal nanometric structures. Finally, the use of nanomarkers is illustrated with stochastic fluorescence microscopy for mapping intracellular structures. We also present some of the future perspectives of optical nanoscopy. Keywords: microscopy, imaging, superresolution, nanodetection, biophysics, medical imaging
International Nuclear Information System (INIS)
Bunshah, R.F.
1976-01-01
A number of different techniques which range over several different aspects of materials research are covered in this volume. They are concerned with property evaluation of 4 0 K and below, surface characterization, coating techniques, techniques for the fabrication of composite materials, computer methods, data evaluation and analysis, statistical design of experiments and non-destructive test techniques. Topics covered in this part include internal friction measurements; nondestructive testing techniques; statistical design of experiments and regression analysis in metallurgical research; and measurement of surfaces of engineering materials
Li, Yan; Li, Luchun; Wu, Zhijuan; Wang, Lulu; Wu, Yongzhong; Li, Dairong; Ma, Uiwen; Shao, Jianghe; Yu, Huiqing; Wang, Donglin
2017-07-01
Evidence has shown that both high expression of the ataxia-telangiectasia mutated (ATM) gene and glioma stem cells (GSCs) are responsible for radioresistance in glioma. Thus, we hypothesized that brain tumor radiosensitivity may be enhanced via silencing of the ATM gene in GSCs. In the present study we successfully induced GSCs from two cell lines and used CD133 and nestin to identify GSCs. A lentivirus was used to deliver siRNA-ATMPuro (A group) to GSCs prior to radiation, while siRNA-HKPuro (N group) and GSCs (C group) were used as negative and blank controls, respectively. RT-qPCR and western blotting were performed to verify the efficiency of the siRNA-ATM technique. The expression of the ATM gene and ATM protein were significantly downregulated post-transfection. Cell Counting Kit-8 (CCK-8) and colony formation assays revealed that the A group demonstrated weak cell proliferation and lower survival fractions post-irradiation compared to the C/N groups. Flow cytometry was used to examine the percentage of cell apoptosis and G2 phase arrest, which were both higher in the A group than in the C/N groups. We found that the comet tail percentage evaluated by comet assay was higher in the A group than in the C/N groups. After radiation treatment, three radiosensitive genes [p53, proliferating cell nuclear antigen (PCNA), survivin] exhibited a decreasing tendency as determined by RT-qPCR. Mice underwent subcutaneous implantation, followed by radiation, and the resulting necrosis and hemorrhage were more obvious in the A group than in the N groups. In conclusion, silencing of ATM via the siRNA technique improved radiosensitivity of GSCs both in vitro and in vivo.
Separation techniques: Chromatography
Coskun, Ozlem
2016-01-01
Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion. Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography. Column chromatography is one of the most common methods of protein purification. PMID:28058406
New techniques in quality assurance
International Nuclear Information System (INIS)
Fornicola, J.C.
1987-01-01
GPU Nuclear Corp. has a multifaceted quality assurance (QA) program. This program includes a comprehensive QA organization to help ensure its implementation. The QA organization employs various techniques in assuring quality at GPU Nuclear. These techniques not only include the typical QA/quality-control verification activities, i.e., QA engineering, quality control, and audits, but also include some new innovative techniques. Several new techniques have been developed for verifying activities. These techniques include monitoring and functional audits of safety systems. Several new techniques for assessing performance and adequacy and effectiveness of plant and QA programs, such as plant assessments and QA systems engineering evaluations, have also been developed. This paper provides an overview of these and other new techniques being employed by GPU Nuclear's QA organization
Microalgae harvesting techniques: A review.
Singh, Gulab; Patidar, S K
2018-07-01
Microalgae with wide range of commercial applications have attracted a lot of attention of the researchers in the last few decades. However, microalgae utilization is not economically sustainable due to high cost of harvesting. A wide range of solid - liquid separation techniques are available for microalgae harvesting. The techniques include coagulation and flocculation, flotation, centrifugation and filtration or a combination of various techniques. Despite the importance of harvesting to the economics and energy balance, there is no universal harvesting technique for microalgae. Therefore, this review focuses on assessing technical, economical and application potential of various harvesting techniques so as to allow selection of an appropriate technology for cost effectively harvesting of microalgae from their culture medium. Various harvesting and concentrating techniques of microalgae were reviewed to suggest order of suitability of the techniques for four main microalgae applications i.e biofuel, human and animal food, high valued products, and water quality restoration. For deciding the order of suitability, a comparative analysis of various harvesting techniques based on the six common criterions (i.e biomass quality, cost, biomass quantity, processing time, species specific and toxicity) has been done. Based on the order of various techniques vis-a-vis various criteria and preferred order of criteria for various applications, order of suitability of harvesting techniques for various applications has been decided. Among various harvesting techniques, coagulation and flocculation, centrifugation and filtration were found to be most suitable for considered applications. These techniques may be used alone or in combination for increasing the harvesting efficiency. Copyright © 2018 Elsevier Ltd. All rights reserved.
Ivezic, Nenad; Potok, Thomas E.
2003-09-30
A method for automatically evaluating a manufacturing technique comprises the steps of: receiving from a user manufacturing process step parameters characterizing a manufacturing process; accepting from the user a selection for an analysis of a particular lean manufacturing technique; automatically compiling process step data for each process step in the manufacturing process; automatically calculating process metrics from a summation of the compiled process step data for each process step; and, presenting the automatically calculated process metrics to the user. A method for evaluating a transition from a batch manufacturing technique to a lean manufacturing technique can comprise the steps of: collecting manufacturing process step characterization parameters; selecting a lean manufacturing technique for analysis; communicating the selected lean manufacturing technique and the manufacturing process step characterization parameters to an automatic manufacturing technique evaluation engine having a mathematical model for generating manufacturing technique evaluation data; and, using the lean manufacturing technique evaluation data to determine whether to transition from an existing manufacturing technique to the selected lean manufacturing technique.
2008-01-01
pathogens also have been detected and identified by numerous nucleic acid-based techniques, including Southern blot hybridization and direct nucleotide...Xanthomonas axonopodis pv. vitians FB-1309; ATCC19320 Lettuce ; USA 1 Xanthomonas axonopodis sp. alfalfae XA129; Med. 2 Alfalfa; Sudan 1 Xanthomonas
Non-contact ultrasound techniques
International Nuclear Information System (INIS)
Khazali Mohd Zin
2001-01-01
Non-contact ultrasound plays significant role in material characterisation and inspection. Unlike conventional ultrasonic techniques, non-contact ultrasonic is mostly applicable to areas where the former has its weaknesses and limitations. It is interesting to note that the non-contact ultrasonic technique has an important significant application in industry. The technique is signified by the fact that the object to be inspected is further away from the ultrasonic source, no couplant is needed and inconsistent pressure between the transducer and the specimen can be eliminated. The paper discusses some of the non-contact ultrasound technique and its applications. (Author)
Principles of fluorescence techniques
2016-01-01
Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.
Disagreements around techniques
International Nuclear Information System (INIS)
Sachsse, H.
The modern man lives in disagreement with its techniques. Technical progress is the foundation of its material and spiritual existence, but he does not like it. Specialization, difficult understanding of the technical procedures and the price that has to be paid for technical progress, have led to a deep-going enstrangement. Therefore the question: What kind of human function has our technique. It is understood as an extension and perfection of our bodily organs, in order to increase our ability to see and to hear enormously, and to intervene formatively in the existing reality. Technique is therefore an instrument of comprehensive self-realization of man. (orig./GL) [de
Advanced analytical techniques
International Nuclear Information System (INIS)
Mrochek, J.E.; Shumate, S.E.; Genung, R.K.; Bahner, C.T.; Lee, N.E.; Dinsmore, S.R.
1976-01-01
The development of several new analytical techniques for use in clinical diagnosis and biomedical research is reported. These include: high-resolution liquid chromatographic systems for the early detection of pathological molecular constituents in physiologic body fluids; gradient elution chromatography for the analysis of protein-bound carbohydrates in blood serum samples, with emphasis on changes in sera from breast cancer patients; electrophoretic separation techniques coupled with staining of specific proteins in cellular isoenzymes for the monitoring of genetic mutations and abnormal molecular constituents in blood samples; and the development of a centrifugal elution chromatographic technique for the assay of specific proteins and immunoglobulins in human blood serum samples
Changes in urological surgical techniques
Directory of Open Access Journals (Sweden)
Oktay Üçer
2010-06-01
Full Text Available Recently, laparoscopic and afterwards robotic techniques have constituted most of urologic surgery procedures. Open surgery may give place to robotic surgery due to possible widespread use of robots in the future. Studies, that compare these two techniques are usually designed about radical prostatectomy, since it is the most common operation performed by using these techniques. In literature,robotic surgery seems more advantageous than other techniques but the most important disadvantage of this technique is cost-effective problems. In present review,history of open, laparoscopic and robotic surgery, and comparison of advantages, disadvantages and cost of these techniques have been discussed with literature.
Al Motowa, Saeed; Al Zobidi, Mohammed
2015-01-01
To illustrate amniotic membrane transplant (AMT) with a novel surgical technique ("sandwich technique") for treating patients with malignant Mooren's ulcer. A case report of a patient with bilateral, malignant Mooren's ulcer who had undergone systemic steroid therapy and topical immunosuppresive therapy to stabilize his condition. However, perforation of cornea occurred in one eye. AMT with a new surgical technique ("Motowa's sandwich technique") was performed to treat this case. On the 1(st) day postoperatively, there was no pain, no photophobia, and visual acuity was same as preoperatively. At 4 weeks postoperatively, visual acuity improved in the right eye to 20/160 with pinhole, and there was no fluorescein staining. The right eye was quiet. Amniotic membrane transplant with "Motowa's sandwich technique" resulted in the preservation of the anatomical integrity and progression of disease was halted along with an improved vision. This technique is a novel surgical modality in treating Mooren's ulcer. Further study on a large cohort of patients is required for evidence-based data to verify the outcome of this initial case report.
Enseignement technique/Technical Training
1998-01-01
Bureautique et techniques administratives/Office Automation & Administrative Techniques - Premiers pas/Getting started - E-Mail - Management Tools - WWW - Applications bureautiques/Office applications
Huang, Hongsheng; Soutyrine, Andrei; Rendulich, Jasmine; O'Rourke, Katherine; Balachandran, Aru
2011-01-01
Tissues unsuitable for standard immunohistochemical and histopathological examinations for chronic wasting disease (CWD) in cervids and for scrapie in sheep are frequently submitted for testing. This study investigated the effects of experimental autolysis on the detection of abnormal prion protein (PrPsc) in lymphoid and central nervous system (CNS) tissues from elk and sheep. The PrPsc was detected using a Western blotting (WB) test following PrPsc enrichment using sodium phosphotungstic acid (PTA) precipitation (PTA-WB). A commercial enzyme-linked immunosorbent assay (ELISA) was used as a reference test for quantitative measurement. This study showed that the amount of PrPsc in lymphoid and CNS tssues from elk and sheep decreased gradually as a result of autolysis, but PrPsc was still detectable after 5 and 15 d incubation at 37°C by PTA-WB for all lymphoid and CNS samples. The results of the ELISA supported those of PTA-WB, particularly for CNS tissues. In conclusion, autolysis at 37°C for 15 d would not significantly affect the detection of PrPsc in lymphoid and CNS tissues by WB and ELISA and, particularly, PTA-WB is a valuable and alternative confirmatory test to detect PrPsc in autolyzed lymphoid and CNS samples.
Tracer techniques in microelectronics
International Nuclear Information System (INIS)
Flachowsky, J.; Freyer, K.
1981-01-01
Tracer technique and neutron activation analysis are capable of measuring impurities in semiconductor material or on the semiconductor surface in a very low concentration range. The methods, combined with autoradiography, are also suitable to determine dopant distributions in silicon. However, both techniques suffer from certain inherent experimental difficulties and/or limitations which are discussed. Methods of tracer technique practicable in the semiconductor field are described. (author)
DEFF Research Database (Denmark)
Burleigh, S.H.
2001-01-01
had high reproducibility and reflected trends in gene expression as observed by Northern blotting. Using this technique, it was demonstrated that both the high-affinity phosphate transporter MtPt2 and a putative nitrate transporter from Medicago truncatula were down-regulated in roots when colonized...
Contamination Control Techniques
Energy Technology Data Exchange (ETDEWEB)
EBY, J.L.
2000-05-16
Welcome to a workshop on contamination Control techniques. This work shop is designed for about two hours. Attendee participation is encouraged during the workshop. We will address different topics within contamination control techniques; present processes, products and equipment used here at Hanford and then open the floor to you, the attendees for your input on the topics.
Contamination Control Techniques
International Nuclear Information System (INIS)
EBY, J.L.
2000-01-01
Welcome to a workshop on contamination Control techniques. This work shop is designed for about two hours. Attendee participation is encouraged during the workshop. We will address different topics within contamination control techniques; present processes, products and equipment used here at Hanford and then open the floor to you, the attendees for your input on the topics
Next generation initiation techniques
Warner, Tom; Derber, John; Zupanski, Milija; Cohn, Steve; Verlinde, Hans
1993-01-01
Four-dimensional data assimilation strategies can generally be classified as either current or next generation, depending upon whether they are used operationally or not. Current-generation data-assimilation techniques are those that are presently used routinely in operational-forecasting or research applications. They can be classified into the following categories: intermittent assimilation, Newtonian relaxation, and physical initialization. It should be noted that these techniques are the subject of continued research, and their improvement will parallel the development of next generation techniques described by the other speakers. Next generation assimilation techniques are those that are under development but are not yet used operationally. Most of these procedures are derived from control theory or variational methods and primarily represent continuous assimilation approaches, in which the data and model dynamics are 'fitted' to each other in an optimal way. Another 'next generation' category is the initialization of convective-scale models. Intermittent assimilation systems use an objective analysis to combine all observations within a time window that is centered on the analysis time. Continuous first-generation assimilation systems are usually based on the Newtonian-relaxation or 'nudging' techniques. Physical initialization procedures generally involve the use of standard or nonstandard data to force some physical process in the model during an assimilation period. Under the topic of next-generation assimilation techniques, variational approaches are currently being actively developed. Variational approaches seek to minimize a cost or penalty function which measures a model's fit to observations, background fields and other imposed constraints. Alternatively, the Kalman filter technique, which is also under investigation as a data assimilation procedure for numerical weather prediction, can yield acceptable initial conditions for mesoscale models. The
Managerial Techniques in Educational Administration.
Lane, John J.
1983-01-01
Management techniques developed during the past 20 years assume the rational bureaucratic model. School administration requires contingent techniques. Quality Circle, Theory Z, and the McKenzie 7-Framework are discussed as techniques to increase school productivity. (MD)
Ma2 antibodies: an evaluation of commercially available detection methods.
Johannis, Wibke; Renno, Joerg H; Wielckens, Klaus; Voltz, Raymond
2011-01-01
Ma2 antibodies belong to the onconeuronal antibodies which define a "definite" paraneoplastic neurological syndrome (PNS). Because of the clinical relevance, use of two separate methods (indirect immunofluorescence technique--IFT--and immunoblot) is advocated; however, with an increasing number of commercially available assay systems, usually only one assay is performed. We compared IFT and three commercially available immunoblots (ravo Diagnostika, Euroimmun, Milenia Biotec) on sera from 35 patients with clinically suspected PNS. 17 were Ma2 antibody associated as defined by consensus result (showing positive reactivity in 2 assays), 18 were Ma2 antibody negative controls. Sensitivity/specificity for single assays were for IFT 94%/94%, for ravo Diagnostika PNS blot 88%/100%, for Euroimmun Neuronal Antigens Profile blot 100%/89%, and for Milenia Biotec MTR blot 94%/100%. Our data confirm, although all tests performed well, a combination of 2 independent assays is still advisable for Ma2 antibody detection in order to achieve higher sensitivity and specificity rates.
Directory of Open Access Journals (Sweden)
Sanjay Kalra
2013-01-01
Full Text Available The forum for injection techniques, India recommendation, the first ever in the country on insulin injcetion techniques, have covered the science and the art of insulin injection technique in an exhaustive manner. However, a few gaps were identified in the document, which are addressed in the current addendum. This article focuses on insulin injection technique in special clinical situations, including geriatric people, women in pregnancy and those with dermatological or surgical disease who live with diabetes. The addendum also covers salient features of administration of insulin using the insulin pump.
Parachute technique for partial penectomy
Directory of Open Access Journals (Sweden)
Fernando Korkes
2010-04-01
Full Text Available PURPOSE: Penile carcinoma is a rare but mutilating malignancy. In this context, partial penectomy is the most commonly applied approach for best oncological results. We herein propose a simple modification of the classic technique of partial penectomy, for better cosmetic and functional results. TECHNIQUE: If partial penectomy is indicated, the present technique can bring additional benefits. Different from classical technique, the urethra is spatulated only ventrally. An inverted "V" skin flap with 0.5 cm of extension is sectioned ventrally. The suture is performed with vicryl 4-0 in a "parachute" fashion, beginning from the ventral portion of the urethra and the "V" flap, followed by the "V" flap angles and than by the dorsal portion of the penis. After completion of the suture, a Foley catheter and light dressing are placed for 24 hours. CONCLUSIONS: Several complex reconstructive techniques have been previously proposed, but normally require specific surgical abilities, adequate patient selection and staged procedures. We believe that these reconstructive techniques are very useful in some specific subsets of patients. However, the technique herein proposed is a simple alternative that can be applied to all men after a partial penectomy, and takes the same amount of time as that in the classic technique. In conclusion, the "parachute" technique for penile reconstruction after partial amputation not only improves the appearance of the penis, but also maintains an adequate function.
Handbook of laboratory techniques
International Nuclear Information System (INIS)
2002-01-01
The Nuclear Regulatory Authority in Argentina have laboratories of support to regulations functions on radiological and nuclear safety, safeguards and physical protection, that have a surface of 2950 m 2 in the Ezeiza Atomic Center. The manual describes in seven chapters the different techniques developed and applied in the laboratories along four decades of existence. The chapter 1: Dedicated to the treatment of environmental samples, described the procedures associated with the different types of samples: deposits, waters, sediments, vegetables, milk, fish and diet. The chapter 2: Details 48 radiochemical techniques associated to the measurements of americium 241, carbon 16, strontium 90, iodine 129, plutonium, radium 226, radon, uranium, nickel and actinides. The chapter 3: Describes the measurements techniques of alpha and gamma spectrometry. The different techniques of biological and physical dosimetry are described in the chapters 5 and 6 respectively. The final chapter is dedicated the techniques of external and internal contamination. It s important to emphasize that this manual contains the standardized technologies that the Nuclear Regulatory Authority of Argentina submits regularly to international comparisons
Ultrasonic techniques for fluids characterization
Povey, Malcolm J W
1997-01-01
This book is a comprehensive and practical guide to the use of ultrasonic techniques for the characterization of fluids. Focusing on ultrasonic velocimetry, the author covers the basic topics and techniques necessaryfor successful ultrasound measurements on emulsions, dispersions, multiphase media, and viscoelastic/viscoplastic materials. Advanced techniques such as scattering, particle sizing, and automation are also presented. As a handbook for industrial and scientific use, Ultrasonic Techniques for Fluids Characterization is an indispensable guide to chemists and chemical engineers using ultrasound for research or process monitoring in the chemical, food processing, pharmaceutical, cosmetic, biotechnology,and fuels industries. Key Features * Appeals to anyone using ultrasound to study fluids * Provides the first detailed description of the ultrasound profiling technique for dispersions * Describes new techniques for measuring phase transitions and nucleation, such as water/ice and oil/fat * Presents the l...
Baas, F.; Bikker, H.; Geurts van Kessel, A.; Melsert, R.; Pearson, P. L.; de Vijlder, J. J.; van Ommen, G. J.
1985-01-01
The human thyroglobulin (Tg) gene is localized to chromosome 8 and regionally to band q24 as shown independently by both in situ hybridization techniques and Southern blot analysis of human-rodent somatic cell hybrids. Analysis of hybrids derived from a Burkitt lymphoma, with a translocation
Directory of Open Access Journals (Sweden)
Lorena Vázquez-Iglesias
2017-06-01
Full Text Available Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.
Nuclear techniques in industry
International Nuclear Information System (INIS)
Hammad, F.H.
1994-01-01
Nuclear techniques are utilized in almost every industry. The discussion in this paper includes discussions on tracer methods and uses nucleonic control systems technology; non-destructive testing techniques and radiation technology. 1 fig., 2 tabs
Airway Clearance Techniques (ACTs)
Full Text Available ... many challenges, including medical, social, and financial. By learning more about how you can manage your disease every day, you can ultimately help find a ... Cycle of Breathing Technique Airway Clearance Techniques Autogenic ...
Approximation techniques for engineers
Komzsik, Louis
2006-01-01
Presenting numerous examples, algorithms, and industrial applications, Approximation Techniques for Engineers is your complete guide to the major techniques used in modern engineering practice. Whether you need approximations for discrete data of continuous functions, or you''re looking for approximate solutions to engineering problems, everything you need is nestled between the covers of this book. Now you can benefit from Louis Komzsik''s years of industrial experience to gain a working knowledge of a vast array of approximation techniques through this complete and self-contained resource.
A Model of Risk Analysis in Analytical Methodology for Biopharmaceutical Quality Control.
Andrade, Cleyton Lage; Herrera, Miguel Angel De La O; Lemes, Elezer Monte Blanco
2018-01-01
One key quality control parameter for biopharmaceutical products is the analysis of residual cellular DNA. To determine small amounts of DNA (around 100 pg) that may be in a biologically derived drug substance, an analytical method should be sensitive, robust, reliable, and accurate. In principle, three techniques have the ability to measure residual cellular DNA: radioactive dot-blot, a type of hybridization; threshold analysis; and quantitative polymerase chain reaction. Quality risk management is a systematic process for evaluating, controlling, and reporting of risks that may affects method capabilities and supports a scientific and practical approach to decision making. This paper evaluates, by quality risk management, an alternative approach to assessing the performance risks associated with quality control methods used with biopharmaceuticals, using the tool hazard analysis and critical control points. This tool provides the possibility to find the steps in an analytical procedure with higher impact on method performance. By applying these principles to DNA analysis methods, we conclude that the radioactive dot-blot assay has the largest number of critical control points, followed by quantitative polymerase chain reaction, and threshold analysis. From the analysis of hazards (i.e., points of method failure) and the associated method procedure critical control points, we conclude that the analytical methodology with the lowest risk for performance failure for residual cellular DNA testing is quantitative polymerase chain reaction. LAY ABSTRACT: In order to mitigate the risk of adverse events by residual cellular DNA that is not completely cleared from downstream production processes, regulatory agencies have required the industry to guarantee a very low level of DNA in biologically derived pharmaceutical products. The technique historically used was radioactive blot hybridization. However, the technique is a challenging method to implement in a quality
Airway Clearance Techniques (ACTs)
Full Text Available ... decisions about your health care. CF Genetics: The Basics CF Mutations Video Series Find Out More About ... of Breathing Technique Airway Clearance Techniques Autogenic Drainage Basics of Lung Care Chest Physical Therapy Coughing and ...
Airway Clearance Techniques (ACTs)
Full Text Available ... Active Cycle of Breathing Technique Airway Clearance Techniques Autogenic Drainage Basics of Lung Care Chest Physical Therapy ... clearance. Facebook Twitter Email More Related Content Medications Autogenic Drainage Positive Expiratory Pressure High-Frequency Chest Wall ...
Airway Clearance Techniques (ACTs)
Full Text Available ... a range of treatment options. Airway Clearance Active Cycle of Breathing Technique Airway Clearance Techniques Autogenic Drainage ... LEGACY GIFT Sponsor a Participant CF Climb CF Cycle for Life Great Strides Xtreme Hike Participate In ...
Airway Clearance Techniques (ACTs)
Full Text Available ... Cycle of Breathing Technique Airway Clearance Techniques Autogenic Drainage Basics of Lung Care Chest Physical Therapy Coughing ... Facebook Twitter Email More Related Content Medications Autogenic Drainage Positive Expiratory Pressure High-Frequency Chest Wall Oscillation ( ...
Sterility test by radiometric technique
International Nuclear Information System (INIS)
Faruq, Muhammad
1980-01-01
Sterility test of pharmaceuticals can be carried out by the application of pharmacopoeia and radiometric technique. In Indonesia the application of pharmacopoeia technique is carried out through liquid germination for aerobacteria and for fungus and yeast. Radiometric technique is applied to autotrop and heterotrop bacteria. (SMN)
Review of advanced imaging techniques
Directory of Open Access Journals (Sweden)
Yu Chen
2012-01-01
Full Text Available Pathology informatics encompasses digital imaging and related applications. Several specialized microscopy techniques have emerged which permit the acquisition of digital images ("optical biopsies" at high resolution. Coupled with fiber-optic and micro-optic components, some of these imaging techniques (e.g., optical coherence tomography are now integrated with a wide range of imaging devices such as endoscopes, laparoscopes, catheters, and needles that enable imaging inside the body. These advanced imaging modalities have exciting diagnostic potential and introduce new opportunities in pathology. Therefore, it is important that pathology informaticists understand these advanced imaging techniques and the impact they have on pathology. This paper reviews several recently developed microscopic techniques, including diffraction-limited methods (e.g., confocal microscopy, 2-photon microscopy, 4Pi microscopy, and spatially modulated illumination microscopy and subdiffraction techniques (e.g., photoactivated localization microscopy, stochastic optical reconstruction microscopy, and stimulated emission depletion microscopy. This article serves as a primer for pathology informaticists, highlighting the fundamentals and applications of advanced optical imaging techniques.
DCT-based cyber defense techniques
Amsalem, Yaron; Puzanov, Anton; Bedinerman, Anton; Kutcher, Maxim; Hadar, Ofer
2015-09-01
With the increasing popularity of video streaming services and multimedia sharing via social networks, there is a need to protect the multimedia from malicious use. An attacker may use steganography and watermarking techniques to embed malicious content, in order to attack the end user. Most of the attack algorithms are robust to basic image processing techniques such as filtering, compression, noise addition, etc. Hence, in this article two novel, real-time, defense techniques are proposed: Smart threshold and anomaly correction. Both techniques operate at the DCT domain, and are applicable for JPEG images and H.264 I-Frames. The defense performance was evaluated against a highly robust attack, and the perceptual quality degradation was measured by the well-known PSNR and SSIM quality assessment metrics. A set of defense techniques is suggested for improving the defense efficiency. For the most aggressive attack configuration, the combination of all the defense techniques results in 80% protection against cyber-attacks with PSNR of 25.74 db.
DEFF Research Database (Denmark)
Hansen, Anders T; Lukacova, Slavka; Lassen-Ramshad, Yasmin A.
2015-01-01
When standard conformal x-ray technique for craniospinal irradiation is used, it is a challenge to achieve satisfactory dose coverage of the target including the area of the cribriform plate, while sparing organs at risk. We present a new intensity-modulated radiation therapy (IMRT), noncoplanar...... patient using the noncoplanar IMRT-based technique, a coplanar IMRT-based technique, and a coplanar volumetric-modulated arch therapy (VMAT) technique. Dosimetry data for all patients were compared with the corresponding data from the conventional treatment plans. The new noncoplanar IMRT technique...... substantially reduced the mean dose to organs at risk compared with the standard radiation technique. The 2 other coplanar techniques also reduced the mean dose to some of the critical organs. However, this reduction was not as substantial as the reduction obtained by the noncoplanar technique. Furthermore...
Airway Clearance Techniques (ACTs)
Full Text Available ... to loosen mucus from airway walls. See how different airway clearance techniques work to help you clear the thick, sticky mucus ... Offer their tips for fitting ACTs into daily life Airway Clearance Techniques | Webcast ... Facebook Twitter ...
Stöggl, Thomas; Müller, Erich; Lindinger, Stefan
2008-09-01
The aims of the study were to: (1) adapt the "double-push" technique from inline skating to cross-country skiing; (2) compare this new skiing technique with the conventional skate skiing cross-country technique; and (3) test the hypothesis that the double-push technique improves skiing speed in a short sprint. 13 elite skiers performed maximum-speed sprints over 100 m using the double-push skate skiing technique and using the conventional "V2" skate skiing technique. Pole and plantar forces, knee angle, cycle characteristics, and electromyography of nine lower body muscles were analysed. We found that the double-push technique could be successfully transferred to cross-country skiing, and that this new technique is faster than the conventional skate skiing technique. The double-push technique was 2.9 +/- 2.2% faster (P push technique had a longer cycle length and a lower cycle rate, and it was characterized by higher muscle activity, higher knee extension amplitudes and velocities, and higher peak foot forces, especially in the first phase of the push-off. Also, the foot was more loaded laterally in the double-push technique than in the conventional skate skiing technique.
Legal technique: approaches to section on types
Directory of Open Access Journals (Sweden)
І. Д. Шутак
2015-11-01
Full Text Available Legal technique is a branch of knowledge about the rules of doing legal work and creating in the process a variety of legal documents, which had previously been part of the theory of law. In modern conditions of the legal technique are isolated in a separate branch of legal science, focused on solving practical problems. The purpose of this article is to analyze the types of legal techniques, in particular, on the basis of theoretical propositions about legal technique to allocate substantial characteristics and types of legal technique. O. Malko and M. Matuzov consider legal technique as a set of rules, techniques, methods of preparation, creation, registration of legal documents, their classification and accounting for their excellence, efficient use. A similar meaning is investing in this concept Alekseev, determining that the legal technique is a set of tools and techniques used in accordance with accepted rules in the formulation and systematization of legal acts to ensure their perfection. So, legal technique – theoretical and applied legal science, which studies the regularities of rational legal practice in the creation, interpretation and implementation of law. In relation to the type of legal techniques in the literature proposed different classifications. For example, G. Muromtsev technique, which is used only in the field of law, divide on the technique of law-making (legislative technique, technique of law enforcement, interpretation, technique of judicial speech, interrogation, notarial activities. V. Kartashov shared legal technique on law making and enforcement (prorealtime, interpretive yourself and prevacidrebatezw, judicial or investigative, prosecutorial, and the like. Some authors clearly indicate that the criterion by which to distinguish types of legal techniques. So, S. Alekseev notes that legal technique is classified from the point of view of the legal nature of the act made on: a techniques of legal acts; b the
Schötta, Anna-Margarita; Wijnveld, Michiel; Stockinger, Hannes; Stanek, Gerold
2017-07-01
Ticks transmit a large number of pathogens capable of causing human disease. In this study, the PCR-reverse line blot (RLB) method was used to screen for pathogens in a total of 554 Ixodes ricinus ticks collected from all provinces of Austria. These pathogens belong to the genera Borrelia , Rickettsiae , Anaplasma / Ehrlichia (including " Candidatus Neoehrlichia"), Babesia , and Coxiella The pathogens with the highest detected prevalence were spirochetes of the Borrelia burgdorferi sensu lato complex, in 142 ticks (25.6%). Borrelia afzelii (80/142) was the most frequently detected species, followed by Borrelia burgdorferi sensu stricto (38/142) and Borrelia valaisiana (36/142). Borrelia garinii/Borrelia bavariensis , Borrelia lusitaniae , and Borrelia spielmanii were found in 28 ticks, 5 ticks, and 1 tick, respectively. Rickettsia spp. were detected in 93 ticks (16.8%): R. helvetica (39/93), R. raoultii (38/93), R. monacensis (2/93), and R. slovaca (1/93). Thirteen Rickettsia samples remain uncharacterized. " Candidatus Neoehrlichia mikurensis," Babesia spp. ( B. venatorum , B. divergens , B. microti ), and Anaplasma phagocytophilum were found in 4.5%, 2.7%, and 0.7%, respectively. Coxiella burnetii was not detected. Multiple microorganisms were detected in 40 ticks (7.2%), and the cooccurrence of Babesia spp. and " Candidatus Neoehrlichia mikurensis" showed a significant positive correlation. We also compared different PCR-RLBs for detection of Borrelia burgdorferi sensu lato and Rickettsia spp. and showed that different detection approaches provide highly diverse results, indicating that analysis of environmental samples remains challenging. IMPORTANCE This study determined the wide spectrum of tick-borne bacterial and protozoal pathogens that can be encountered in Austria. Surveillance of (putative) pathogenic microorganisms occurring in the environment is of medical importance, especially when those agents can be transmitted by ticks and cause disease. The
Analytical system availability techniques
Brouwers, J.J.H.; Verbeek, P.H.J.; Thomson, W.R.
1987-01-01
Analytical techniques are presented to assess the probability distributions and related statistical parameters of loss of production from equipment networks subject to random failures and repairs. The techniques are based on a theoretical model for system availability, which was further developed
Differential effects of gastric bypass and banding on the ...
African Journals Online (AJOL)
Data collected included changes of body mass index (BMI), blood pressure, fasting blood sugar, fasting serum insulin, insulin resistance (HOMA-IR) and lipid profile. In addition, apo A-IV was determined by the Western blot technique. Results: The results demonstrated a highly significant reduction in body weight as ...
Weterman, MAJ; Janssen, [No Value; Janssen, HAP; vandenBerg, E; Fisher, SE; Craig, [No Value; vanKessel, AG
1996-01-01
A combination of Southern blot analysis on a panel of tumor-derived somatic cell hybrids and fluorescence in situ hybridization techniques was used to map YACs, cosmids and DNA markers from the Xp11.2 region relative to the X chromosome breakpoint of the renal cell carcinoma-associated
GPU Pro advanced rendering techniques
Engel, Wolfgang
2010-01-01
This book covers essential tools and techniques for programming the graphics processing unit. Brought to you by Wolfgang Engel and the same team of editors who made the ShaderX series a success, this volume covers advanced rendering techniques, engine design, GPGPU techniques, related mathematical techniques, and game postmortems. A special emphasis is placed on handheld programming to account for the increased importance of graphics on mobile devices, especially the iPhone and iPod touch.Example programs and source code can be downloaded from the book's CRC Press web page.
EDITORIAL: Measurement techniques for multiphase flows Measurement techniques for multiphase flows
Okamoto, Koji; Murai, Yuichi
2009-11-01
Research on multiphase flows is very important for industrial applications, including power stations, vehicles, engines, food processing and so on. Multiphase flows originally have nonlinear features because of multiphase systems. The interaction between the phases plays a very interesting role in the flows. The nonlinear interaction causes the multiphase flows to be very complicated. Therefore techniques for measuring multiphase flows are very useful in helping to understand the nonlinear phenomena. The state-of-the-art measurement techniques were presented and discussed at the sixth International Symposium on Measurement Techniques for Multiphase Flows (ISMTMF2008) held in Okinawa, Japan, on 15-17 December 2008. This special feature of Measurement Science and Technology includes selected papers from ISMTMF2008. Okinawa has a long history as the Ryukyus Kingdom. China, Japan and many western Pacific countries have had cultural and economic exchanges through Okinawa for over 1000 years. Much technical and scientific information was exchanged at the symposium in Okinawa. The proceedings of ISMTMF2008 apart from these special featured papers were published in Journal of Physics: Conference Series vol. 147 (2009). We would like to express special thanks to all the contributors to the symposium and this special feature. This special feature will be a milestone in measurement techniques for multiphase flows.
Wang, He; Xiao, Meng; Kong, Fanrong; Chen, Sharon; Dou, Hong-Tao; Sorrell, Tania; Li, Ruo-Yu; Xu, Ying-Chun
2011-01-01
Eleven reference and 25 clinical isolates of Fusarium were subject to multilocus DNA sequence analysis to determine the species and haplotypes of the fusarial isolates from Beijing and Shandong, China. Seven loci were analyzed: the translation elongation factor 1 alpha gene (EF-1α); the nuclear rRNA internal transcribed spacer (ITS), large subunit (LSU), and intergenic spacer (IGS) regions; the second largest subunit of the RNA polymerase gene (RPB2); the calmodulin gene (CAM); and the mitochondrial small subunit (mtSSU) rRNA gene. We also evaluated an IGS-targeted PCR/reverse line blot (RLB) assay for species/haplotype identification of Fusarium. Twenty Fusarium species and seven species complexes were identified. Of 25 clinical isolates (10 species), the Gibberella (Fusarium) fujikuroi species complex was the commonest (40%) and was followed by the Fusarium solani species complex (FSSC) (36%) and the F. incarnatum-F. equiseti species complex (12%). Six FSSC isolates were identified to the species level as FSSC-3+4, and three as FSSC-5. Twenty-nine IGS, 27 EF-1α, 26 RPB2, 24 CAM, 18 ITS, 19 LSU, and 18 mtSSU haplotypes were identified; 29 were unique, and haplotypes for 24 clinical strains were novel. By parsimony informative character analysis, the IGS locus was the most phylogenetically informative, and the rRNA gene regions were the least. Results by RLB were concordant with multilocus sequence analysis for all isolates. Amphotericin B was the most active drug against all species. Voriconazole MICs were high (>8 μg/ml) for 15 (42%) isolates, including FSSC. Analysis of larger numbers of isolates is required to determine the clinical utility of the seven-locus sequence analysis and RLB assay in species classification of fusaria. PMID:21389150
Amselem, S; Nunes, V; Vidaud, M; Estivill, X; Wong, C; d'Auriol, L; Vidaud, D; Galibert, F; Baiget, M; Goossens, M
1988-01-01
We have delineated the molecular lesions causing beta-thalassemia in Spain, a country that has witnessed the passage of different Mediterranean populations over the centuries, in order to evaluate the extent of heterogeneity of these mutations and to make possible simplified prenatal diagnosis of the disorder in that country. The use of the polymerase chain-reaction (PCR) technique to preferentially amplify beta-globin DNA sequences that contain the most frequent beta-thalassemia mutations in...
Ethical Use of Gestalt Techniques.
Given, Jane A.
The purpose of this paper is to engender a healthy respect for Gestalt theory and techniques and the use of the techniques in the client's best interest and in the interest of positive professional and self-development in the practitioner. An overview of Gestalt techniques is provided, concentrating on the two category divisions of experiments and…
Advanced in-flight measurement techniques
Lawson, Nicholas; Jentink, Henk; Kompenhans, Jürgen
2013-01-01
The book presents a synopsis of the main results achieved during the 3 year EU-project "Advanced Inflight Measurement Techniques (AIM)" which applied advanced image based measurement techniques to industrial flight testing. The book is intended to be not only an overview on the AIM activities but also a guide on the application of advanced optical measurement techniques for future flight testing. Furthermore it is a useful guide for engineers in the field of experimental methods and flight testing who face the challenge of a future requirement for the development of highly accurate non-intrusive in-flight measurement techniques.
[Impressions techniques--Part 2].
Levartovsky, S; Masri, M; Alter, E; Pilo, R
2012-10-01
A dental impression is a positive replica of the teeth, the surrounding gingiva and the border between them; the purpose of which is to create an accurate master model. Two major techniques for impressions exist today: The conventional and the digital impressions. The current article describes both techniques. In the conventional impressions, it is important to choose a proper tray, stock or custom, and to mix the material properly. The commonly used impression techniques for making a conventional impression are described with a review on the effect of the technique on its accuracy. The effect of the wash bulk on the accuracy of the stone dies and/or the restoration is discussed, as well. The digital impressions with their advantages and disadvantages are described in comparison to the conventional impressions. Although, digital impressions eliminate some of the negative characteristics of conventional impressions, proper soft-tissue management and isolation of tooth preparation margins is still mandatory.
International Nuclear Information System (INIS)
Hoeschen, W.
2003-01-01
The TIG (Tungsten-inert gas) orbital welding technique is applied in all areas of pipe welding. The process is mainly used for austenitic and ferritic materials but also for materials like aluminium, nickel, and titanium alloys are commonly welded according to this technique. Thin-walled as well as thick-walled pipes are welded economically. The application of orbital welding is of particular interest in the area of maintenance of thick-walled pipes that is described in this article. (orig.) [de
Analysis and analytical techniques
Energy Technology Data Exchange (ETDEWEB)
Batuecas Rodriguez, T [Department of Chemistry and Isotopes, Junta de Energia Nuclear, Madrid (Spain)
1967-01-01
The technology associated with the use of organic coolants in nuclear reactors depends to a large extent on the determination and control of their physical and chemical properties, and particularly on the viability, speed, sensitivity, precision and accuracy (depending on the intended usage) of the methods employed in detection and analytical determination. This has led to the study and development of numerous techniques, some specially designed for the extreme conditions involved in working with the types of product in question and others adapted from existing techniques. In the specific case of polyphenyl and hydropolyphenyl mixtures, which have been the principal subjects of study to date and offer greatest promise, the analytical problems are broadly as follows: Composition of initial product or virgin coolant composition of macro components and amounts of organic and inorganic impurities; Coolant during and after operation. Determination of gases and organic compounds produced by pyrolysis and radiolysis (degradation and polymerization products); Control of systems for purifying and regenerating the coolant after use. Dissolved pressurization gases; Detection of intermediate products during decomposition; these are generally very unstable (free radicals); Degree of fouling and film formation. Tests to determine potential formation of films; Corrosion of structural elements and canning materials; Health and safety. Toxicity, inflammability and impurities that can be activated. Although some of the above problems are closely interrelated and entail similar techniques, they vary as to degree of difficulty. Another question is the difficulty of distinguishing clearly between techniques for determining physical and physico-chemical properties, on one hand, and analytical techniques on the other. Any classification is therefore somewhat arbitrary (for example, in the case of dosimetry and techniques for determining mean molecular weights or electrical conductivity
Anatomic partial nephrectomy: technique evolution.
Azhar, Raed A; Metcalfe, Charles; Gill, Inderbir S
2015-03-01
Partial nephrectomy provides equivalent long-term oncologic and superior functional outcomes as radical nephrectomy for T1a renal masses. Herein, we review the various vascular clamping techniques employed during minimally invasive partial nephrectomy, describe the evolution of our partial nephrectomy technique and provide an update on contemporary thinking about the impact of ischemia on renal function. Recently, partial nephrectomy surgical technique has shifted away from main artery clamping and towards minimizing/eliminating global renal ischemia during partial nephrectomy. Supported by high-fidelity three-dimensional imaging, novel anatomic-based partial nephrectomy techniques have recently been developed, wherein partial nephrectomy can now be performed with segmental, minimal or zero global ischemia to the renal remnant. Sequential innovations have included early unclamping, segmental clamping, super-selective clamping and now culminating in anatomic zero-ischemia surgery. By eliminating 'under-the-gun' time pressure of ischemia for the surgeon, these techniques allow an unhurried, tightly contoured tumour excision with point-specific sutured haemostasis. Recent data indicate that zero-ischemia partial nephrectomy may provide better functional outcomes by minimizing/eliminating global ischemia and preserving greater vascularized kidney volume. Contemporary partial nephrectomy includes a spectrum of surgical techniques ranging from conventional-clamped to novel zero-ischemia approaches. Technique selection should be tailored to each individual case on the basis of tumour characteristics, surgical feasibility, surgeon experience, patient demographics and baseline renal function.
Kitchin, C R
2013-01-01
Telescopes and Techniques has proved itself in its first two editions, having become probably one of the most widely used astronomy texts, both for amateur astronomers and astronomy and astrophysics undergraduates. Both earlier editions of the book were widely used for introductory practical astronomy courses in many universities. In this Third Edition the author guides the reader through the mathematics, physics and practical techniques needed to use today's telescopes (from the smaller models to the larger instruments installed in many colleges) and how to find objects in the sky. Most of the physics and engineering involved is described fully and requires little prior knowledge or experience. Both visual and electronic imaging techniques are covered, together with an introduction to how data (measurements) should be processed and analyzed. A simple introduction to radio telescopes is also included. Brief coverage of the more advanced topics of photometry and spectroscopy are included, but mainly to enable ...
Directory of Open Access Journals (Sweden)
Serkan AKKOYUNLU
2013-08-01
Conclusion: Exposure with response prevention is a basic and effective technique. Every cognitive behavior therapist must be able to implement this technique and be cognizant of pearls of this procedure. (Journal of Cognitive Behavioral Psychotherapy and Research 2013, 2: 121-128 [JCBPR 2013; 2(2.000: 121-128
Techniques of radiation dosimetry
International Nuclear Information System (INIS)
Mahesk, K.
1985-01-01
A text and reference with an interdisciplinary approach to physics, atomic energy, radiochemistry, and radiobiology. Chapters examine basic principles, experimental techniques, the methodology of dose experiments, and applications. Treats 14 different dosimetric techniques, including ionization chamber, thermoluminescence, and lyoluminescence. Considers the conceptual aspects and characteristic features of radiation
Zhang, She-Hong; Huang, Qian
2013-12-01
Pancreatic cancer is a highly aggressive malignant tumor. In the present study, we performed several methods, including CCK-8 assay, immunofluorescence technique, western blotting and flow cytometry, to determine the effects of VP16 (etoposide) on Panc-1 pancreatic cancer cells. The results demonstrated that VP16 inhibited the growth of and induced apoptosis in Panc-1 cells. Western blot analysis showed that VP16 inhibited the expression of Bcl-2 and enhanced the expression of Bax, caspases-3 and -9, cytochrome c and PARP. Notably, a strong inhibitory effect of VP16 on Panc-1 cells mainly occurred in non-CSCs. These data provide a new strategy for the therapy of pancreatic cancer.
DNA fingerprinting of spore-forming bacterial isolates, using Bacillus ...
African Journals Online (AJOL)
User
Full Length Research Paper ... resulted in a search for better techniques for classifying ... only a few laboratories worldwide are able to perform a ... MATERIALS AND METHODS. Bacterial ... s with distilled water and blotted dry with tissue paper (Kimberly- ... A test on the quality and quantity of DNA extracted was conducted.
Adaptive algebraic reconstruction technique
International Nuclear Information System (INIS)
Lu Wenkai; Yin Fangfang
2004-01-01
Algebraic reconstruction techniques (ART) are iterative procedures for reconstructing objects from their projections. It is proven that ART can be computationally efficient by carefully arranging the order in which the collected data are accessed during the reconstruction procedure and adaptively adjusting the relaxation parameters. In this paper, an adaptive algebraic reconstruction technique (AART), which adopts the same projection access scheme in multilevel scheme algebraic reconstruction technique (MLS-ART), is proposed. By introducing adaptive adjustment of the relaxation parameters during the reconstruction procedure, one-iteration AART can produce reconstructions with better quality, in comparison with one-iteration MLS-ART. Furthermore, AART outperforms MLS-ART with improved computational efficiency
Millimicrosecond pulse techniques
Lewis, Ian A D
1959-01-01
Millimicrosecond Pulse Techniques, Second Edition focuses on millimicrosecond pulse techniques and the development of devices of large bandwidth, extending down to comparatively low frequencies (1 Mc/s). Emphasis is on basic circuit elements and pieces of equipment of universal application. Specific applications, mostly in the field of nuclear physics instrumentation, are considered. This book consists of eight chapters and opens with an introduction to some of the terminology employed by circuit engineers as well as theoretical concepts, including the laws of circuit analysis, Fourier analysi
Combination of behavioural and cognitive techniques
Directory of Open Access Journals (Sweden)
Leonida Rotvejn Pajič
2001-03-01
Full Text Available The cognitive - behavioral approach in the treatment of behavioral disorders, emotional disorders and learning difficulties is not limited to a single theoretical principle or technique, but integrates different techniques. The variants techniques are derived from theoretical and empirical findings in different domains of developmental psychology (self-control, social cognition, memory, metacognitive and attritional processes. The aim of the article is to present some basic cognitive and behavioral techniques that are individually adapted during therapy and mutually complete each other. A practical case of combining techniques is presented.
van Til, Janine; Groothuis-Oudshoorn, Catharina; Lieferink, Marijke; Dolan, James; Goetghebeur, Mireille
2014-01-01
There is an increased interest in the use of multi-criteria decision analysis (MCDA) to support regulatory and reimbursement decision making. The EVIDEM framework was developed to provide pragmatic multi-criteria decision support in health care, to estimate the value of healthcare interventions, and to aid in priority-setting. The objectives of this study were to test 1) the influence of different weighting techniques on the overall outcome of an MCDA exercise, 2) the discriminative power in weighting different criteria of such techniques, and 3) whether different techniques result in similar weights in weighting the criteria set proposed by the EVIDEM framework. A sample of 60 Dutch and Canadian students participated in the study. Each student used an online survey to provide weights for 14 criteria with two different techniques: a five-point rating scale and one of the following techniques selected randomly: ranking, point allocation, pairwise comparison and best worst scaling. The results of this study indicate that there is no effect of differences in weights on value estimates at the group level. On an individual level, considerable differences in criteria weights and rank order occur as a result of the weight elicitation method used, and the ability of different techniques to discriminate in criteria importance. Of the five techniques tested, the pair-wise comparison of criteria has the highest ability to discriminate in weights when fourteen criteria are compared. When weights are intended to support group decisions, the choice of elicitation technique has negligible impact on criteria weights and the overall value of an innovation. However, when weights are used to support individual decisions, the choice of elicitation technique influences outcome and studies that use dissimilar techniques cannot be easily compared. Weight elicitation through pairwise comparison of criteria is preferred when taking into account its superior ability to discriminate between
The Delphi Technique in Educational Research
Directory of Open Access Journals (Sweden)
Ravonne A. Green
2014-04-01
Full Text Available The Delphi Technique has been useful in educational settings in forming guidelines, standards, and in predicting trends. Judd lists these major uses of the Delphi Technique in higher education: (a cost-effectiveness, (b cost–benefit analysis, (c curriculum and campus planning, and (d university-wide educational goals and objectives. The thorough Delphi researcher seeks to reconcile the Delphi consensus with current literature, institutional research, and the campus environment. This triangle forms a sound base for responsible research practice. This book gives an overview of the Delphi Technique and the primary uses of this technique in research. This article on the Delphi Technique will give the researcher an invaluable resource for learning about the Delphi Technique and for applying this method in educational research projects.
Relaxation techniques for stress
... raise your heart rate. This is called the stress response. Relaxation techniques can help your body relax and lower your blood pressure ... also many other types of breathing techniques you can learn. In many cases, you do not need much ... including those that cause stress. Meditation has been practiced for thousands of years, ...
International Nuclear Information System (INIS)
Berquist, T.H.; Bender, C.E.; James, E.M.; Brown, M.L.; McLeod, R.A.; Broderick, D.F.; Welch, T.J.
1989-01-01
Proper application of imaging procedures is essential to obtain needed information for diagnosis and therapy planning in patients with suspected foot and/or ankle pathology. This paper provides basic background data for the numerous imaging techniques
Strongly correlated systems experimental techniques
Mancini, Ferdinando
2015-01-01
The continuous evolution and development of experimental techniques is at the basis of any fundamental achievement in modern physics. Strongly correlated systems (SCS), more than any other, need to be investigated through the greatest variety of experimental techniques in order to unveil and crosscheck the numerous and puzzling anomalous behaviors characterizing them. The study of SCS fostered the improvement of many old experimental techniques, but also the advent of many new ones just invented in order to analyze the complex behaviors of these systems. Many novel materials, with functional properties emerging from macroscopic quantum behaviors at the frontier of modern research in physics, chemistry and materials science, belong to this class of systems. The volume presents a representative collection of the modern experimental techniques specifically tailored for the analysis of strongly correlated systems. Any technique is presented in great detail by its own inventor or by one of the world-wide recognize...
Two sampling techniques for game meat
Directory of Open Access Journals (Sweden)
Maretha van der Merwe
2013-03-01
Full Text Available A study was conducted to compare the excision sampling technique used by the export market and the sampling technique preferred by European countries, namely the biotrace cattle and swine test. The measuring unit for the excision sampling was grams (g and square centimetres (cm2 for the swabbing technique. The two techniques were compared after a pilot test was conducted on spiked approved beef carcasses (n = 12 that statistically proved the two measuring units correlated. The two sampling techniques were conducted on the same game carcasses (n = 13 and analyses performed for aerobic plate count (APC, Escherichia coli and Staphylococcus aureus, for both techniques. A more representative result was obtained by swabbing and no damage was caused to the carcass. Conversely, the excision technique yielded fewer organisms and caused minor damage to the carcass. The recovery ratio from the sampling technique improved 5.4 times for APC, 108.0 times for E. coli and 3.4 times for S. aureus over the results obtained from the excision technique. It was concluded that the sampling methods of excision and swabbing can be used to obtain bacterial profiles from both export and local carcasses and could be used to indicate whether game carcasses intended for the local market are possibly on par with game carcasses intended for the export market and therefore safe for human consumption.
Two sampling techniques for game meat.
van der Merwe, Maretha; Jooste, Piet J; Hoffman, Louw C; Calitz, Frikkie J
2013-03-20
A study was conducted to compare the excision sampling technique used by the export market and the sampling technique preferred by European countries, namely the biotrace cattle and swine test. The measuring unit for the excision sampling was grams (g) and square centimetres (cm2) for the swabbing technique. The two techniques were compared after a pilot test was conducted on spiked approved beef carcasses (n = 12) that statistically proved the two measuring units correlated. The two sampling techniques were conducted on the same game carcasses (n = 13) and analyses performed for aerobic plate count (APC), Escherichia coli and Staphylococcus aureus, for both techniques. A more representative result was obtained by swabbing and no damage was caused to the carcass. Conversely, the excision technique yielded fewer organisms and caused minor damage to the carcass. The recovery ratio from the sampling technique improved 5.4 times for APC, 108.0 times for E. coli and 3.4 times for S. aureus over the results obtained from the excision technique. It was concluded that the sampling methods of excision and swabbing can be used to obtain bacterial profiles from both export and local carcasses and could be used to indicate whether game carcasses intended for the local market are possibly on par with game carcasses intended for the export market and therefore safe for human consumption.
Informationization nuclear apparatus communication technique
International Nuclear Information System (INIS)
Yu Tiqi; Fang Zongliang; Wen Qilin
2006-01-01
The paper explains the request of communication ability in nuclear technique application area. Based on the actuality of nuclear apparatus communication ability, and mainly combining with the development of communication technique, the authors analyzes the application trend of communication technique applying in nuclear apparatus, for the apparatus and system needing communication ability, they need selecting suitable communication means to make them accomplish the task immediately and effectively. (authors)
Applying Brainstorming Techniques to EFL Classroom
Toshiya, Oishi; 湘北短期大学; aPart-time Lecturer at Shohoku College
2015-01-01
This paper focuses on brainstorming techniques for English language learners. From the author's teaching experiences at Shohoku College during the academic year 2014-2015, the importance of brainstorming techniques was made evident. The author explored three elements of brainstorming techniques for writing using literaturereviews: lack of awareness, connecting to prior knowledge, and creativity. The literature reviews showed the advantage of using brainstorming techniques in an English compos...
Enseignement technique : Séminaire de l'enseigement technique
2004-01-01
Mardi 4 mai 2004 SEMINAIRE DE L'ENSEIGNEMENT TECHNIQUE de 14:00 à 17:00 - Training Centre Auditorium - bât. 593 Le SPC - Statistical Process Control - dans une démarche de qualité totale Hakim Bourahla, Maurice Arrius, Charbel Tannous - ABW Concept - F-74950 SCIONZIER, France Ce nouveau séminaire de l'Enseignement Technique sera consacré à la présentation du SPC, le Statistical Process Control. Le SPC apporte une grande efficacité dans l'amélioration de la qualité des produits. Cette méthode, permettant d'assurer une qualité optimum à l'aide d'un outil statistique, est fondée sur deux concepts de base : le suivi et le pilotage des procédés industriels par cartes de contrôle, et l'étude des capabilités des systèmes de production. - Origine et objectifs du SPC - Concepts du ...
Enseignement technique : Séminaire de l'enseignement technique
2004-01-01
Mardi 4 mai 2004 SEMINAIRE DE L'ENSEIGNEMENT TECHNIQUE de 14:00 à 17:00 - Training Centre Auditorium - bât. 593 Le SPC - Statistical Process Control - dans une démarche de qualité totale Hakim Bourahla, Maurice Arrius, Charbel Tannous - ABW Concept - F-74950 SCIONZIER, France Ce nouveau séminaire de l'Enseignement Technique sera consacré à la présentation du SPC, le Statistical Process Control. Le SPC apporte une grande efficacité dans l'amélioration de la qualité des produits. Cette méthode, permettant d'assurer une qualité optimum à l'aide d'un outil statistique, est fondée sur deux concepts de base : le suivi et le pilotage des procédés industriels par cartes de contrôle, et l'étude des capabilités des systèmes de production. - Origine et objectifs du SPC - Concepts du ...
Hyphenated analytical techniques for materials characterisation
International Nuclear Information System (INIS)
Armstrong, Gordon; Kailas, Lekshmi
2017-01-01
This topical review will provide a survey of the current state of the art in ‘hyphenated’ techniques for characterisation of bulk materials, surface, and interfaces, whereby two or more analytical methods investigating different properties are applied simultaneously to the same sample to better characterise the sample than can be achieved by conducting separate analyses in series using different instruments. It is intended for final year undergraduates and recent graduates, who may have some background knowledge of standard analytical techniques, but are not familiar with ‘hyphenated’ techniques or hybrid instrumentation. The review will begin by defining ‘complementary’, ‘hybrid’ and ‘hyphenated’ techniques, as there is not a broad consensus among analytical scientists as to what each term means. The motivating factors driving increased development of hyphenated analytical methods will also be discussed. This introduction will conclude with a brief discussion of gas chromatography-mass spectroscopy and energy dispersive x-ray analysis in electron microscopy as two examples, in the context that combining complementary techniques for chemical analysis were among the earliest examples of hyphenated characterisation methods. The emphasis of the main review will be on techniques which are sufficiently well-established that the instrumentation is commercially available, to examine physical properties including physical, mechanical, electrical and thermal, in addition to variations in composition, rather than methods solely to identify and quantify chemical species. Therefore, the proposed topical review will address three broad categories of techniques that the reader may expect to encounter in a well-equipped materials characterisation laboratory: microscopy based techniques, scanning probe-based techniques, and thermal analysis based techniques. Examples drawn from recent literature, and a concluding case study, will be used to explain the
Hyphenated analytical techniques for materials characterisation
Armstrong, Gordon; Kailas, Lekshmi
2017-09-01
This topical review will provide a survey of the current state of the art in ‘hyphenated’ techniques for characterisation of bulk materials, surface, and interfaces, whereby two or more analytical methods investigating different properties are applied simultaneously to the same sample to better characterise the sample than can be achieved by conducting separate analyses in series using different instruments. It is intended for final year undergraduates and recent graduates, who may have some background knowledge of standard analytical techniques, but are not familiar with ‘hyphenated’ techniques or hybrid instrumentation. The review will begin by defining ‘complementary’, ‘hybrid’ and ‘hyphenated’ techniques, as there is not a broad consensus among analytical scientists as to what each term means. The motivating factors driving increased development of hyphenated analytical methods will also be discussed. This introduction will conclude with a brief discussion of gas chromatography-mass spectroscopy and energy dispersive x-ray analysis in electron microscopy as two examples, in the context that combining complementary techniques for chemical analysis were among the earliest examples of hyphenated characterisation methods. The emphasis of the main review will be on techniques which are sufficiently well-established that the instrumentation is commercially available, to examine physical properties including physical, mechanical, electrical and thermal, in addition to variations in composition, rather than methods solely to identify and quantify chemical species. Therefore, the proposed topical review will address three broad categories of techniques that the reader may expect to encounter in a well-equipped materials characterisation laboratory: microscopy based techniques, scanning probe-based techniques, and thermal analysis based techniques. Examples drawn from recent literature, and a concluding case study, will be used to explain the
Microscopy techniques in flavivirus research.
Chong, Mun Keat; Chua, Anthony Jin Shun; Tan, Terence Tze Tong; Tan, Suat Hoon; Ng, Mah Lee
2014-04-01
The Flavivirus genus is composed of many medically important viruses that cause high morbidity and mortality, which include Dengue and West Nile viruses. Various molecular and biochemical techniques have been developed in the endeavour to study flaviviruses. However, microscopy techniques still have irreplaceable roles in the identification of novel virus pathogens and characterization of morphological changes in virus-infected cells. Fluorescence microscopy contributes greatly in understanding the fundamental viral protein localizations and virus-host protein interactions during infection. Electron microscopy remains the gold standard for visualizing ultra-structural features of virus particles and infected cells. New imaging techniques and combinatory applications are continuously being developed to push the limit of resolution and extract more quantitative data. Currently, correlative live cell imaging and high resolution three-dimensional imaging have already been achieved through the tandem use of optical and electron microscopy in analyzing biological specimens. Microscopy techniques are also used to measure protein binding affinities and determine the mobility pattern of proteins in cells. This chapter will consolidate on the applications of various well-established microscopy techniques in flavivirus research, and discuss how recently developed microscopy techniques can potentially help advance our understanding in these membrane viruses. Copyright © 2013 Elsevier Ltd. All rights reserved.
Teaching Speaking Through Debate Technique
Directory of Open Access Journals (Sweden)
. Suranto
2016-07-01
Full Text Available Abstract : Teaching Speaking Through Debate Technique. Speaking is one of the basic competence from the other fourth basic competence (listening, speaking, reading and writing. Speaking ability should be mastered by every students, in order to achieve that competence students should be given the right technique to study sepaking. The successfull of the students speaking can be seen from their ability to express idea, thought and feeling through speaking. The objective of this Action Research is to improve students’s oral communication skill through the debate technique. This study was conducted at MA Ma’arif Nu 5 Sekampung Lampung Timur from March to April 2014. The research data were taken from students in the eleventh class, with 28 students and analyzed qualitatively and quantitatively. The research findings indicate that there are improvements in students’ english speaking skill through the debate technique. By analyzing data qualitatively and quantitatively from the end of the first cycle to the second cycle and it was found that the students’ English speaking skill increased 20,9% over the standard that has been determined by the researcher that is 65%. The researcher concludes that the students’ english speaking skill can be improve through the debate technique in learning process. Key words : action research, debate technique, english speaking skill
Highly reliable TOFD UT Technique
International Nuclear Information System (INIS)
Acharya, G.D.; Trivedi, S.A.R.; Pai, K.B.
2003-01-01
The high performance of the time of flight diffraction technique (TOFD) with regard to the detection capabilities of weld defects such as crack, slag, lack of fusion has led to a rapidly increasing acceptance of the technique as a pre?service inspection tool. Since the early 1990s TOFD has been applied to several projects, where it replaced the commonly used radiographic testing. The use of TOM lead to major time savings during new build and replacement projects. At the same time the TOFD technique was used as base line inspection, which enables monitoring in the future for critical welds, but also provides documented evidence for life?time. The TOFD technique as the ability to detect and simultaneously size flows of nearly any orientation within the weld and heat affected zone. TOM is recognized as a reliable, proven technique for detection and sizing of defects and proven to be a time saver, resulting in shorter shutdown periods and construction project times. Thus even in cases where inspection price of TOFD per welds is higher, in the end it will result in significantly lower overall costs and improve quality. This paper deals with reliability, economy, acceptance criteria and field experience. It also covers comparative study between radiography technique Vs. TOFD. (Author)
Lymphography - an outdated technique
International Nuclear Information System (INIS)
Peters, P.E.
1982-01-01
The indications for lymphography have changed with the availability of non-invasive techniques like ultrasonic techniques and computerized tomography. This review discusses: Recent results of lymphography in histologically verified patient collectives with lymphatic systemic diseases and lymphatic metastizing tumors. The present role of lymphography is derived from this status report as well as the future perspectives. (orig.) [de
International Nuclear Information System (INIS)
Froejmark, M.
1992-10-01
The report presents a wide, easily understandable description of presentation technique and man-machine communication. General fundamentals for the man-machine interface are illustrated, and the factors that affect the interface are described. A model is presented for describing the operators work situation, based on three different levels in the operators behaviour. The operator reacts routinely in the face of simple, known problems, and reacts in accordance with predetermined plans in the face of more complex, recognizable problems. Deep fundamental knowledge is necessary for truly complex questions. Today's technical status and future development have been studied. In the future, the operator interface will be based on standard software. Functions such as zooming, integration of video pictures, and sound reproduction will become common. Video walls may be expected to come into use in situations in which several persons simultaneously need access to the same information. A summary of the fundamental rules for the design of good picture ergonomics and design requirements for control rooms are included in the report. In conclusion, the report describes a presentation technique within the Distribution Automation and Demand Side Management area and analyses the know-how requirements within Vattenfall. If different systems are integrated, such as geographical information systems and operation monitoring systems, strict demands are made on the expertise of the users for achieving a user-friendly technique which is matched to the needs of the human being. (3 figs.)
Higher-order techniques in computational electromagnetics
Graglia, Roberto D
2016-01-01
Higher-Order Techniques in Computational Electromagnetics explains 'high-order' techniques that can significantly improve the accuracy, computational cost, and reliability of computational techniques for high-frequency electromagnetics, such as antennas, microwave devices and radar scattering applications.
International Nuclear Information System (INIS)
Saur, S; Frengen, J; Fjellsboe, L M B; Lindmo, T
2009-01-01
The contralateral breast (CLB) doses for three tangential techniques were characterized by using a female thorax phantom and GafChromic EBT film. Dose calculations by the pencil beam and collapsed cone algorithms were included for comparison. The film dosimetry reveals a highly inhomogeneous dose distribution within the CLB, and skin doses due to the medial fields that are several times higher than the interior dose. These phenomena are not correctly reproduced by the calculation algorithms. All tangential techniques were found to give a mean CLB dose of approximately 0.5 Gy. All wedged fields resulted in higher CLB doses than the corresponding open fields, and the lateral open fields resulted in higher CLB doses than the medial open fields. More than a twofold increase in the mean CLB dose from the medial open field was observed for a 90 deg. change of the collimator orientation. Replacing the physical wedge with a virtual wedge reduced the mean dose to the CLB by 35% and 16% for the medial and lateral fields, respectively. Lead shielding reduced the skin dose for a tangential technique by approximately 50%, but the mean CLB dose was only reduced by approximately 11%. Finally, a technique based on open medial fields in combination with several IMRT fields is proposed as a technique for minimizing the CLB dose. With and without lead shielding, the mean CLB dose using this technique was found to be 0.20 and 0.27 Gy, respectively.
The Matador Technique: A technique to improve prostatic ...
African Journals Online (AJOL)
A. Pai
Abstract. The accuracy of brachytherapy seed implantation is reduced by the movement of the prostate when needles are introduced transperineally. This report describes a simple method of introducing the first two needles, which reduces prostate deflection. This technique is analogous to the way a matador uses two ...
MODIFIED TECHNIQUE OF TOTAL LARYNGECTOMY
Directory of Open Access Journals (Sweden)
Predrag Spirić
2010-12-01
Full Text Available Surgical technique of total laryngectomy is well presented in many surgical textbooks. Essentially, it has remained the same since Gluck an Soerensen in 1922 described all its details. Generally, it stresses the U shape skin incision with releasing laryngeal structures and removing larynx from up to down. Further, pharyngeal reconstruction is performed with different kinds of sutures in two or more layers and is finished with skin suture and suction drainage. One of worst complications following this surgery is pharyngocutaneous fistula (PF. Modifications proposed in this this article suggests vertical skin incision with larynx removal from below upwards. In pharyngeal reconstruction we used the running locked suture in submucosal plan with „tobacco sac“ at the end on the tongue base instead of traditional T shaped suture. Suction drains were not used.The aim of study was to present the modified surgical technique of total laryingectomy and its impact on hospital stay duration and pharyngocutanous fistula formation. In this randomized study we analyzed 49 patients operated with modified surgical technique compared to 49 patient operated with traditional surgical technique of total laryngectomy. The modified technique of total laryngectomy was presented. Using modified technique we managed to decrease the PF percentage from previous 20,41% to acceptable 8,16% (p=0,0334. Also, the average hospital stay was shortened from 14,96 to 10,63 days (t =-2.9850; p=0.0358.The modified technique of total laryngectomy is safe, short and efficient surgical intervention which decreases the number of pharyngocutaneos fistulas and shortens the hospital stay.
A Comparative of business process modelling techniques
Tangkawarow, I. R. H. T.; Waworuntu, J.
2016-04-01
In this era, there is a lot of business process modeling techniques. This article is the research about differences of business process modeling techniques. For each technique will explain about the definition and the structure. This paper presents a comparative analysis of some popular business process modelling techniques. The comparative framework is based on 2 criteria: notation and how it works when implemented in Somerleyton Animal Park. Each technique will end with the advantages and disadvantages. The final conclusion will give recommend of business process modeling techniques that easy to use and serve the basis for evaluating further modelling techniques.
International Nuclear Information System (INIS)
Smith, S.W.; Kruger, J.
1985-06-01
Radioisotope handling procedures followed by personnel of the Nuclear Techniques Industrial Service (NTIS) during the conduction of investigations in industry are described. Possible radiological implications as a result of the various measuring techniques and different types of plants are discussed. Conditions under which permanent authorization has been granted for the use of radioisotopes are mentioned
Response of Various Tomato Genotypes to Begomovirus Infection and Its Improved Diagnostic
Directory of Open Access Journals (Sweden)
NOOR AIDAWATI
2007-09-01
Full Text Available Begomovirus infection was identified from tomato growing areas in West Java (Bogor, Central Java (Boyolali, and D.I. Yogyakarta (Kaliurang. Efforts to reduce the infection among others are planting resistance varieties. This research was undertaken to evaluate 14 tomato genotypes for their response to the infection. Dot blot hybridization using nonradioactive (digoxigenin DNA probe was employed to determine the presence of begomovirus in inoculated plants. Polymerase chain reaction-amplified product of DNA clone of tobacco leaf curl virus –Indonesia was used as a source of DNA probe. All of tomato genotypes evaluated in this study was infected separately by three strain of begomovirus (GVPSlm, GVABy, GVCBgr. Tomato genotypes Bonanza, Jelita, Safira, Permata, Presto, PSPT 8, PSPT 5B, Apel-Belgia, Karibia, Mitra, PSPT 9, Marta, and PSPT 2, showed susceptible or highly susceptible response to the three strains of begomovirus. Exception to those was shown by cv. Intan which resulted in moderate resistance when inoculated with GVCBgr although it resulted susceptible response with the other two strains. Dot-blot hybridization technique was proved to be a powerful tool to detect begomovirus infection in plants showing symptom as well as symptom-less plants. Accumulation of the virus in those plants was relatively high, except in cv. Bonanza and Apel-Belgia. Dot-blot hybridization technique using DIG-labeled DNA probe was able to detect begomovirus DNA in infected tissue up to 10−2 dilution factor.
Progress in diagnosis of viral hepatitis A, B, C, D and E
International Nuclear Information System (INIS)
Kurstak, E.; Hossain, A.; Kurstak, C.
1996-01-01
The effective use of new molecular biological techniques towards the reliable diagnosis of HCV and other viral liver infections has been updated in this review. The applications PCR techniques with amplification of reverse transcribed cDNA seems to prove an effective means for assaying HCV infections. A very recent one-stage PCR assay of HCV RNA combined with either liquid hybridization or Southern blot analysis, equal in sensitivity to the nested PCR assay but with sharply reduced potential for contamination appears to be promising. Future further development of reliable and automated RT-PCR assay would be particularly interesting for diagnosis of HCV infections. PCR apparently remains the most useful test for the appraisal of HBV infection in sera-negative patients with liver disease. It has now made possible the confirmation of observations made with the spot blot or Southern blot test and provided access to the nucleotide sequence analysis of these viral mutant forms. The rapidity and simplicity of these viral forms. The rapidity and simplicity of the the newly developed latex agglutination method using ISTA also makes it a viable alternative for the determination of HBsAg. Cloning of HEV, sequencing of the viral genome and expression of recombinant HEV proteins has undoubtedly facilitated significant progress in the development of methods for identification of HEV infection in patients. Recently the availability of specific, more sensitive assays as recombinant-based EIAs has made the diagnosis of hepatitis E very much practicable. (author)
Composite Techniques Based Color Image Compression
Directory of Open Access Journals (Sweden)
Zainab Ibrahim Abood
2017-03-01
Full Text Available Compression for color image is now necessary for transmission and storage in the data bases since the color gives a pleasing nature and natural for any object, so three composite techniques based color image compression is implemented to achieve image with high compression, no loss in original image, better performance and good image quality. These techniques are composite stationary wavelet technique (S, composite wavelet technique (W and composite multi-wavelet technique (M. For the high energy sub-band of the 3rd level of each composite transform in each composite technique, the compression parameters are calculated. The best composite transform among the 27 types is the three levels of multi-wavelet transform (MMM in M technique which has the highest values of energy (En and compression ratio (CR and least values of bit per pixel (bpp, time (T and rate distortion R(D. Also the values of the compression parameters of the color image are nearly the same as the average values of the compression parameters of the three bands of the same image.
Recent developments in building diagnosis techniques
2016-01-01
This book presents a collection of recent research on building diagnosis techniques related to construction pathology, hygrothermal behavior and durability, and diagnostic techniques. It highlights recent advances and new developments in the field of building physics, building anomalies in materials and components, new techniques for improved energy efficiency analysis, and diagnosis techniques such as infrared thermography. This book will be of interest to a wide readership of professionals, scientists, students, practitioners, and lecturers.
Applicability of statistical process control techniques
Schippers, W.A.J.
1998-01-01
This paper concerns the application of Process Control Techniques (PCTs) for the improvement of the technical performance of discrete production processes. Successful applications of these techniques, such as Statistical Process Control Techniques (SPC), can be found in the literature. However, some
Glycoprotein and proteoglycan techniques
International Nuclear Information System (INIS)
Beeley, J.G.
1985-01-01
The aim of this book is to describe techniques which can be used to answer some of the basic questions about glycosylated proteins. Methods are discussed for isolation, compositional analysis, and for determination of the primary structure of carbohydrate units and the nature of protein-carbohydrate linkages of glycoproteins and proteoglycans. High resolution NMR is considered, as well as radioactive labelling techniques. (Auth.)
Laser induced pyrolysis techniques
International Nuclear Information System (INIS)
Vanderborgh, N.E.
1976-01-01
The application of laser pyrolysis techniques to the problems of chemical analysis is discussed. The processes occurring during laser pyrolysis are first briefly reviewed. The problems encountered in laser pyrolysis gas chromatography are discussed using the analysis of phenanthrene and binary hydrocarbons. The application of this technique to the characterization of naturally occurring carbonaceous material such as oil shales and coal is illustrated
International Nuclear Information System (INIS)
Ogawa, Kazuhiro; Amao, Satoshi; Ichikawa, Yuji; Shoji, Tetsuo
2008-01-01
This study proposes an innovative technique for repairing of cracked or damaged parts of structures, such as nuclear or thermal power plants, by means of cold gas dynamic spray (CS) technique. In the case of generation of cracks etc. in the structure, the cracks can be repaired by welding. However, the welding spends considerable time on repair, and also needs special skills. The CS technique is known as a new technique not only for coatings but also for thick depositions. It has many advantages, i.e. dense deposition, high deposition rate and low oxidation. Therefore, it has a possibility to apply the CS technique instead of welding to repair the cracks etc. In this study, the cold gas dynamic spray technique as a new repairing technique for some structures is introduced. (author)
Huber, David Miles
2013-01-01
As the most popular and authoritative guide to recording Modern Recording Techniques provides everything you need to master the tools and day to day practice of music recording and production. From room acoustics and running a session to mic placement and designing a studio Modern Recording Techniques will give you a really good grounding in the theory and industry practice. Expanded to include the latest digital audio technology the 7th edition now includes sections on podcasting, new surround sound formats and HD and audio.If you are just starting out or looking for a step up
Walls, JM
2013-01-01
This volume provides a comprehensive and up to the minute review of the techniques used to determine the nature and composition of surfaces. Originally published as a special issue of the Pergamon journal Vacuum, it comprises a carefully edited collection of chapters written by specialists in each of the techniques and includes coverage of the electron and ion spectroscopies, as well as the atom-imaging methods such as the atom probe field ion microscope and the scanning tunnelling microscope. Surface science is an important area of study since the outermost surface layers play a crucial role
International Nuclear Information System (INIS)
Drost, W.
1983-01-01
The single well technique method includes measurement of parameters of groundwater flow in saturated rock. For determination of filtration velocity the dilution of radioactive tracer is measured, for direction logging the collimeter is rotated in the probe linked with the compass. The limiting factor for measurement of high filtration velocities is the occurrence of turbulent flow. The single well technique is used in civil engineering projects, water works and subsurface drainage of liquid waste from disposal sites. The radioactive tracer method for logging the vertical fluid movement in bore-holes is broadly used in groundwater survey and exploitation. (author)
Craniospinal irradiation techniques
Energy Technology Data Exchange (ETDEWEB)
Scarlatescu, Ioana, E-mail: scarlatescuioana@gmail.com; Avram, Calin N. [Faculty of Physics, West University of Timisoara, Bd. V. Parvan 4, 300223 Timisoara (Romania); Virag, Vasile [County Hospital “Gavril Curteanu” - Oradea (Romania)
2015-12-07
In this paper we present one treatment plan for irradiation cases which involve a complex technique with multiple beams, using the 3D conformational technique. As the main purpose of radiotherapy is to administrate a precise dose into the tumor volume and protect as much as possible all the healthy tissues around it, for a case diagnosed with a primitive neuro ectoderm tumor, we have developed a new treatment plan, by controlling one of the two adjacent fields used at spinal field, in a way that avoids the fields superposition. Therefore, the risk of overdose is reduced by eliminating the field divergence.
Authentication techniques for smart cards
International Nuclear Information System (INIS)
Nelson, R.A.
1994-02-01
Smart card systems are most cost efficient when implemented as a distributed system, which is a system without central host interaction or a local database of card numbers for verifying transaction approval. A distributed system, as such, presents special card and user authentication problems. Fortunately, smart cards offer processing capabilities that provide solutions to authentication problems, provided the system is designed with proper data integrity measures. Smart card systems maintain data integrity through a security design that controls data sources and limits data changes. A good security design is usually a result of a system analysis that provides a thorough understanding of the application needs. Once designers understand the application, they may specify authentication techniques that mitigate the risk of system compromise or failure. Current authentication techniques include cryptography, passwords, challenge/response protocols, and biometrics. The security design includes these techniques to help prevent counterfeit cards, unauthorized use, or information compromise. This paper discusses card authentication and user identity techniques that enhance security for microprocessor card systems. It also describes the analysis process used for determining proper authentication techniques for a system
Radio techniques for probing the terrestrial ionosphere.
Hunsucker, R. D.
The subject of the book is a description of the basic principles of operation, plus the capabilities and limitations of all generic radio techniques employed to investigate the terrestrial ionosphere. The purpose of this book is to present to the reader a balanced treatment of each technique so they can understand how to interpret ionospheric data and decide which techniques are most effective for studying specific phenomena. The first two chapters outline the basic theory underlying the techniques, and each following chapter discusses a separate technique. This monograph is entirely devoted to techniques in aeronomy and space physics. The approach is unique in its presentation of the principles, capabilities and limitations of the most important presently used radio techniques. Typical examples of data are shown for the various techniques, and a brief historical account of the technique development is presented. An extended annotated bibliography of the salient papers in the field is included.
Two sampling techniques for game meat
van der Merwe, Maretha; Jooste, Piet J.; Hoffman, Louw C.; Calitz, Frikkie J.
2013-01-01
A study was conducted to compare the excision sampling technique used by the export market and the sampling technique preferred by European countries, namely the biotrace cattle and swine test. The measuring unit for the excision sampling was grams (g) and square centimetres (cm2) for the swabbing technique. The two techniques were compared after a pilot test was conducted on spiked approved beef carcasses (n = 12) that statistically proved the two measuring units correlated. The two sampling...
A True Open-Loop Synchronization Technique
DEFF Research Database (Denmark)
Golestan, Saeed; Vidal, Ana; Yepes, Alejandro G.
2016-01-01
Synchronization techniques can be broadly classified into two major categories: Closed-loop and open-loop methods. The open-loop synchronization (OLS) techniques, contrary to the closed-loop ones, are unconditionally stable and benefit from a fast dynamic response. Their performance, however, tends...... is to develop a true OLS (and therefore, unconditionally stable) technique without any need for the calculation of sine and cosine functions. The effectiveness of the proposed synchronization technique is confirmed through the simulation and experimental results....
Technique Selectively Represses Immune System
... Research Matters December 3, 2012 Technique Selectively Represses Immune System Myelin (green) encases and protects nerve fibers (brown). A new technique prevents the immune system from attacking myelin in a mouse model of ...
Nuclear techniques in analytical chemistry
Moses, Alfred J; Gordon, L
1964-01-01
Nuclear Techniques in Analytical Chemistry discusses highly sensitive nuclear techniques that determine the micro- and macro-amounts or trace elements of materials. With the increasingly frequent demand for the chemical determination of trace amounts of elements in materials, the analytical chemist had to search for more sensitive methods of analysis. This book accustoms analytical chemists with nuclear techniques that possess the desired sensitivity and applicability at trace levels. The topics covered include safe handling of radioactivity; measurement of natural radioactivity; and neutron a
Directory of Open Access Journals (Sweden)
Zlatan Saračević
2011-09-01
Full Text Available Due to the complexity of the motion, shot put technique is described in phases for easier analysis, easer learning of technique and error correction. It is complete so that in its implementation the transition from phase to phase is not noticed. In aforementioned and described phases of O'Brian spinal shot put technique a large distance, emptiness and disconnection appear between the initial position phase and a phase of overtaking the device, which in the training methods and training technique in primary and secondary education, as well as for students and athletes beginners in shot put represents a major problem regarding connecting, training and technique advancement. Therefore, this work is aimed at facilitating the methods of training of shot put technique, extending from four to six phases, which have been described and include the complete O'Brian technique.
Point-source inversion techniques
Langston, Charles A.; Barker, Jeffrey S.; Pavlin, Gregory B.
1982-11-01
A variety of approaches for obtaining source parameters from waveform data using moment-tensor or dislocation point source models have been investigated and applied to long-period body and surface waves from several earthquakes. Generalized inversion techniques have been applied to data for long-period teleseismic body waves to obtain the orientation, time function and depth of the 1978 Thessaloniki, Greece, event, of the 1971 San Fernando event, and of several events associated with the 1963 induced seismicity sequence at Kariba, Africa. The generalized inversion technique and a systematic grid testing technique have also been used to place meaningful constraints on mechanisms determined from very sparse data sets; a single station with high-quality three-component waveform data is often sufficient to discriminate faulting type (e.g., strike-slip, etc.). Sparse data sets for several recent California earthquakes, for a small regional event associated with the Koyna, India, reservoir, and for several events at the Kariba reservoir have been investigated in this way. Although linearized inversion techniques using the moment-tensor model are often robust, even for sparse data sets, there are instances where the simplifying assumption of a single point source is inadequate to model the data successfully. Numerical experiments utilizing synthetic data and actual data for the 1971 San Fernando earthquake graphically demonstrate that severe problems may be encountered if source finiteness effects are ignored. These techniques are generally applicable to on-line processing of high-quality digital data, but source complexity and inadequacy of the assumed Green's functions are major problems which are yet to be fully addressed.
Nuclear techniques in agriculture
International Nuclear Information System (INIS)
Bhagwat, S.G.
2012-01-01
Crops provide us food grains and many other products. Demand for food and other agricultural products is increasing. There is also need for improvement of quality of the agricultural produce. There are several technologies in use for achieving the goal of increasing the quantity and quality of agricultural produce. Nuclear techniques provide us with an option which has certain advantages. The characteristics of crop plants are determined by the genetic make up of the plant. Traditionally the genetic make up was modified using conventional breeding techniques such as cross breeding to improve crops for yield, disease resistance, stress tolerance, resistance to insect pests or to improve quality. New varieties of crops are produced which replace the earlier ones and thus the demands are met. The process of development of new varieties is long and time consuming. Nuclear technique called mutation breeding provides an efficient way of breeding new varieties or improving the older ones. This technique merely enhances the process of occurrence of mutations. In nature mutations occur at a rate of approximately one in a million, while when mutations are induced using radiations such as gamma rays the efficiency of inducing mutations is enhanced. Useful mutations are selected, the mutants are evaluated and developed as a new variety. In the Nuclear Agriculture and Biotechnology Division (NA and BTD) this technique has been used to develop mutants of many crop plants. The mutants can be used to develop a variety directly or by using it in further breeding programme. Using these approaches the NA and BTD has developed 40 new varieties of crops such as groundnut, mungbean, urid, pigeon pea, mustard, soybean, sunflower, cowpea, jute. These varieties are developed in collaboration with other agricultural institutions and are popular among the farming community. The method of mutation breeding can be applied to many other crops for improvement. There is increasing interest among
Novel food processing techniques
Directory of Open Access Journals (Sweden)
Vesna Lelas
2006-12-01
Full Text Available Recently, a lot of investigations have been focused on development of the novel mild food processing techniques with the aim to obtain the high quality food products. It is presumed also that they could substitute some of the traditional processes in the food industry. The investigations are primarily directed to usage of high hydrostatic pressure, ultrasound, tribomechanical micronization, microwaves, pulsed electrical fields. The results of the scientific researches refer to the fact that application of some of these processes in particular food industry can result in lots of benefits. A significant energy savings, shortening of process duration, mild thermal conditions, food products with better sensory characteristics and with higher nutritional values can be achieved. As some of these techniques act also on the molecular level changing the conformation, structure and electrical potential of organic as well as inorganic materials, the improvement of some functional properties of these components may occur. Common characteristics of all of these techniques are treatment at ambient or insignificant higher temperatures and short time of processing (1 to 10 minutes. High hydrostatic pressure applied to various foodstuffs can destroy some microorganisms, successfully modify molecule conformation and consequently improve functional properties of foods. At the same time it acts positively on the food products intend for freezing. Tribomechanical treatment causes micronization of various solid materials that results in nanoparticles and changes in structure and electrical potential of molecules. Therefore, the significant improvement of some rheological and functional properties of materials occurred. Ultrasound treatment proved to be potentially very successful technique of food processing. It can be used as a pretreatment to drying (decreases drying time and improves functional properties of food, as extraction process of various components
Deschênes, Philippe; Chano, Frédéric; Dionne, Léa-Laurence; Pittet, Didier; Longtin, Yves
2017-08-01
The efficacy of the World Health Organization (WHO)-recommended handwashing technique against Clostridium difficile is uncertain, and whether it could be improved remains unknown. Also, the benefit of using a structured technique instead of an unstructured technique remains unclear. This study was a prospective comparison of 3 techniques (unstructured, WHO, and a novel technique dubbed WHO shortened repeated [WHO-SR] technique) to remove C difficile. Ten participants were enrolled and performed each technique. Hands were contaminated with 3 × 10 6 colony forming units (CFU) of a nontoxigenic strain containing 90% spores. Efficacy was assessed using the whole-hand method. The relative efficacy of each technique and of a structured (either WHO or WHO-SR) vs an unstructured technique were assessed by Mann-Whitney U test and Wilcoxon signed-rank test. The median effectiveness of the unstructured, WHO, and WHO-SR techniques in log 10 CFU reduction was 1.30 (interquartile range [IQR], 1.27-1.43), 1.71 (IQR, 1.34-1.91), and 1.70 (IQR, 1.54-2.42), respectively. The WHO-SR technique was significantly more efficacious than the unstructured technique (P = .01). Washing hands with a structured technique was more effective than washing with an unstructured technique (median, 1.70 vs 1.30 log 10 CFU reduction, respectively; P = .007). A structured washing technique is more effective than an unstructured technique against C difficile. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.
A review on creatinine measurement techniques.
Mohabbati-Kalejahi, Elham; Azimirad, Vahid; Bahrami, Manouchehr; Ganbari, Ahmad
2012-08-15
This paper reviews the entire recent global tendency for creatinine measurement. Creatinine biosensors involve complex relationships between biology and micro-mechatronics to which the blood is subjected. Comparison between new and old methods shows that new techniques (e.g. Molecular Imprinted Polymers based algorithms) are better than old methods (e.g. Elisa) in terms of stability and linear range. All methods and their details for serum, plasma, urine and blood samples are surveyed. They are categorized into five main algorithms: optical, electrochemical, impedometrical, Ion Selective Field-Effect Transistor (ISFET) based technique and chromatography. Response time, detection limit, linear range and selectivity of reported sensors are discussed. Potentiometric measurement technique has the lowest response time of 4-10 s and the lowest detection limit of 0.28 nmol L(-1) belongs to chromatographic technique. Comparison between various techniques of measurements indicates that the best selectivity belongs to MIP based and chromatographic techniques. Copyright © 2012 Elsevier B.V. All rights reserved.
National Research Council Canada - National Science Library
Pound, John D
1998-01-01
... easy and important refinements often are not published. This much anticipated 2nd edition of Immunochemzcal Protocols therefore aims to provide a user-friendly up-to-date handbook of reliable techniques selected to suit the needs of molecular biologists. It covers the full breadth of the relevant established immunochemical methods, from protein blotting and immunoa...
Statistical Techniques for Project Control
Badiru, Adedeji B
2012-01-01
A project can be simple or complex. In each case, proven project management processes must be followed. In all cases of project management implementation, control must be exercised in order to assure that project objectives are achieved. Statistical Techniques for Project Control seamlessly integrates qualitative and quantitative tools and techniques for project control. It fills the void that exists in the application of statistical techniques to project control. The book begins by defining the fundamentals of project management then explores how to temper quantitative analysis with qualitati
Scalable Techniques for Formal Verification
Ray, Sandip
2010-01-01
This book presents state-of-the-art approaches to formal verification techniques to seamlessly integrate different formal verification methods within a single logical foundation. It should benefit researchers and practitioners looking to get a broad overview of the spectrum of formal verification techniques, as well as approaches to combining such techniques within a single framework. Coverage includes a range of case studies showing how such combination is fruitful in developing a scalable verification methodology for industrial designs. This book outlines both theoretical and practical issue
Artificial intelligence techniques in Prolog
Shoham, Yoav
1993-01-01
Artificial Intelligence Techniques in Prolog introduces the reader to the use of well-established algorithmic techniques in the field of artificial intelligence (AI), with Prolog as the implementation language. The techniques considered cover general areas such as search, rule-based systems, and truth maintenance, as well as constraint satisfaction and uncertainty management. Specific application domains such as temporal reasoning, machine learning, and natural language are also discussed.Comprised of 10 chapters, this book begins with an overview of Prolog, paying particular attention to Prol
Use of communication techniques by Maryland dentists.
Maybury, Catherine; Horowitz, Alice M; Wang, Min Qi; Kleinman, Dushanka V
2013-12-01
Health care providers' use of recommended communication techniques can increase patients' adherence to prevention and treatment regimens and improve patient health outcomes. The authors conducted a survey of Maryland dentists to determine the number and type of communication techniques they use on a routine basis. The authors mailed a 30-item questionnaire to a random sample of 1,393 general practice dentists and all 169 members of the Maryland chapter of the American Academy of Pediatric Dentistry. The overall response rate was 38.4 percent. Analysis included descriptive statistics, analysis of variance and ordinary least squares regression analysis to examine the association of dentists' characteristics with the number of communication techniques used. They set the significance level at P communication techniques and 3.6 of the seven basic techniques, whereas pediatric dentists reported using a mean of 8.4 and 3.8 of those techniques, respectively. General dentists who had taken a communication course outside of dental school were more likely than those who had not to use the 18 techniques (P communication course outside of dental school were more likely than those who had not to use the 18 techniques (P communication techniques that dentists used routinely varied across the 18 techniques and was low for most techniques. Practical Implications. Professional education is needed both in dental school curricula and continuing education courses to increase use of recommended communication techniques. Specifically, dentists and their team members should consider taking communication skills courses and conducting an overall evaluation of their practices for user friendliness.
Lu, Tzong-Shi; Yiao, Szu-Yu; Lim, Kenneth; Jensen, Roderick V; Hsiao, Li-Li
2010-07-01
The identification of differences in protein expression resulting from methodical variations is an essential component to the interpretation of true, biologically significant results. We used the Lowry and Bradford methods- two most commonly used methods for protein quantification, to assess whether differential protein expressions are a result of true biological or methodical variations. MATERIAL #ENTITYSTARTX00026; Differential protein expression patterns was assessed by western blot following protein quantification by the Lowry and Bradford methods. We have observed significant variations in protein concentrations following assessment with the Lowry versus Bradford methods, using identical samples. Greater variations in protein concentration readings were observed over time and in samples with higher concentrations, with the Bradford method. Identical samples quantified using both methods yielded significantly different expression patterns on Western blot. We show for the first time that methodical variations observed in these protein assay techniques, can potentially translate into differential protein expression patterns, that can be falsely taken to be biologically significant. Our study therefore highlights the pivotal need to carefully consider methodical approaches to protein quantification in techniques that report quantitative differences.
Bartholomew, J.L.; Rohovec, J.S.; Fryer, J.L.
1989-01-01
Both monoclonal and polyclonal antisera were produced against Ceratomyxa shasta. Ascites containing trophozoites of the parasite was collected from infected fish and used as antigen for immunization of mice. The resulting monoclonal antibodies reacted specifically with trophozoite and sporoblast stages but did not react with C. shasta spores by either indirect fluorescent antibody techniques or in Western blots. This indicates that some C. shasta antigens are specific to certain life stages of the parasite. Polyclonal antiserum was produced in a rabbit by injecting a spore protein electro-eluted from an SDS-polyacrylamide gel. This antiserum reacted with both trophozoites and spores by indirect fluorescent antibody techniques and in Western blots. All antisera were tested for cross-reactivity to trout white blood cells, a contaminant of the ascites, and to other myxosporea. Two monoclonal antibodies reacted with white blood cells and myxosporea of the genera Sphaerospora and Myxobilatus. One hybridoma produced antibodies of high specificity for C. shasta pre-spore stages. This is the first report of a monoclonal antibody produced against a myxosporean parasite.
New diagnostic technique for Zeeman-compensated atomic beam slowing: technique and results
Molenaar, P.A.; Straten, P. van der; Heideman, H.G.M.; Metcalf, H.
1997-01-01
We have developed a new diagnostic tool for the study of Zeeman-compensated slowing of an alkali atomic beam. Our time-of-flight technique measures the longitudinal veloc- ity distribution of the slowed atoms with a resolution below the Doppler limit of 30 cm/s. Furthermore, it can map the position and velocity distribution of atoms in either ground hyperfine level inside the solenoid without any devices inside the solenoid. The technique reveals the optical pumping ef- fects, and shows in de...
International Nuclear Information System (INIS)
Choi, Sang Woo; Lee, Joon Hyun; Seo, Kyeong Cheol; Byun, Joon Hyung
2005-01-01
Fiber reinforced plastic material should be inspected in fabrication process in order to enhance quality by prevent defects such as delamination and void. Generally, ultrasonic technique is widely used to evaluate FRP. In conventional ultrasonic techniques, transducer should be contacted on FRP. However, conventional contacting method could not be applied in fabrication process and novel non-contact evaluating technique was required. Laser-based ultrasonic technique was tried to evaluate CFRP plate. Laser-based ultrasonic waves propagated on CFRP were received with various transducers such as accelerometer and AE sensor in order to evaluate the properties of waves due to the variation of frequency. Velocities of laser-based ultrasonic waves were evaluated for various fiber orientation. In addition, laser interferometry was used to receive ultrasonic wave in CFRP and frequency was analysed
The Delphi technique in radiography education research
International Nuclear Information System (INIS)
John-Matthews, J.St.; Wallace, M.J.; Robinson, L.
2017-01-01
Objectives: To describe and review the Delphi technique as a tool for radiographers engaged in mixed-methods research whereby agreement is required on the proficiencies needed by educational programmes for pre- and post- registration radiographers. This is achieved through a description offering a brief history of the technique. Through a literature search, radiography education research using this technique is identified. A protocol for a research project using the technique is presented. Using this worked example, advantages and disadvantages of the method are explored including sampling of participants, sample size, number of rounds and methods of feedback. Key findings: There are limited examples of the use of the Delphi technique in radiography literature including considerations on how to select experts and panel size. Conclusion: The Delphi technique is a suitable method for establishing collective agreement in the design of radiography educational interventions. Additional research is needed to deepen this evidence-based knowledge. - Highlights: • The Delphi Technique is used to gain collective agreement in forecasting healthcare education priorities. • There is increase of the use of the technique in healthcare education literature. • There are some examples of the technique in radiography education research. • A worked example of this technique is provided to critically evaluate this tool.
Stargate: Energy Management Techniques
Vijay Raghunathan; Mani Srivastava; Trevor Pering; Roy Want
2004-01-01
This poster presents techniques for energy efficient operation of the Stargate wireless platform. In addition to conventional power management techniques such as dynamic voltage and scaling and processor shutdown, the Stargate features several mechanisms for energy efficient operation of the communication subsystem, such as support for hierarchical radios, Bluetooth based remote wakeup, mote based wakeup, etc. Finally, design optimizations including the use of power gating, and provision for ...
Modern techniques of surface science
Woodruff, D Phil
2016-01-01
This fully revised, updated and reorganised third edition provides a thorough introduction to the characterisation techniques used in surface science and nanoscience today. Each chapter brings together and compares the different techniques used to address a particular research question, including how to determine the surface composition, surface structure, surface electronic structure, surface microstructure at different length scales (down to sub-molecular), and the molecular character of adsorbates and their adsorption or reaction properties. Readers will easily understand the relative strengths and limitations of the techniques available to them and, ultimately, will be able to select the most suitable techniques for their own particular research purposes. This is an essential resource for researchers and practitioners performing materials analysis, and for senior undergraduate students looking to gain a clear understanding of the underlying principles and applications of the different characterisation tec...
Radiation technique for the destruction of plutella
International Nuclear Information System (INIS)
1980-01-01
Plutella can be destroyed by cultivation technique where cabbage is grown interchangebly with other plants, biological technique where the natural enemy of plutella is used, and chemical technique where insecticide is used. Those method do not better results than the sterile male technique which can be either nuclear or chemical in nature. Laboratory, semifield research, and research in a limited field have been carried out when applying the sterile male technique. (SMN)
Experimental techniques; Techniques experimentales
Energy Technology Data Exchange (ETDEWEB)
Roussel-Chomaz, P. [GANIL CNRS/IN2P3, CEA/DSM, 14 - Caen (France)
2007-07-01
This lecture presents the experimental techniques, developed in the last 10 or 15 years, in order to perform a new class of experiments with exotic nuclei, where the reactions induced by these nuclei allow to get information on their structure. A brief review of the secondary beams production methods will be given, with some examples of facilities in operation or under project. The important developments performed recently on cryogenic targets will be presented. The different detection systems will be reviewed, both the beam detectors before the targets, and the many kind of detectors necessary to detect all outgoing particles after the reaction: magnetic spectrometer for the heavy fragment, detection systems for the target recoil nucleus, {gamma} detectors. Finally, several typical examples of experiments will be detailed, in order to illustrate the use of each detector either alone, or in coincidence with others. (author)
Food physics and radiation techniques
International Nuclear Information System (INIS)
Szabo, A. S.
1999-01-01
In the lecture information is given about food physics, which is a rather new, interdisciplinary field of science, connecting food science and applied physics. The topics of radioactivity of foodstuffs and radiation techniques in the food industry are important parts of food physics detailed information will be given about the main fields (e.g. radio stimulation, food preservation) of radiation techniques in the agro-food sector. Finally some special questions of radioactive contamination of foodstuffs in hungary and applicability of radioanalytical techniques (e.g. Inaa) for food investigation will be analyzed and discussed
A simplified indirect bonding technique
Directory of Open Access Journals (Sweden)
Radha Katiyar
2014-01-01
Full Text Available With the advent of lingual orthodontics, indirect bonding technique has become an integral part of practice. It involves placement of brackets initially on the models and then their transfer to teeth with the help of transfer trays. Problems encountered with current indirect bonding techniques used are (1 the possibility of adhesive flash remaining around the base of the brackets which requires removal (2 longer time required for the adhesive to gain enough bond strength for secure tray removal. The new simplified indirect bonding technique presented here overcomes both these problems.
A note on outbreak of cowpea mild mottle virus (CPMMV) in common bean in the River Nile State, Sudan
International Nuclear Information System (INIS)
Hamed, A. A.; Elkhair, J.; Elfaha, A.
2010-01-01
An outbreak of a devastating virus disease occurred in common ben (phaseolus vulgaris) in Berber area, the River Nile State, during the 2004/2005 cropping season, with symptoms of stunting and yellowing. The disease incidence reached a level of more than 85% in all visited fields. One hundred fifty symptomatic samples, collected from different fields at Hudeiba, Berber and Shendi were blotted on nitrocellulose membranes and tested for the presence of different viruses, using the tissue blot immunoassay (TBIA) technique. The results of the serological tests revealed that 95% of the samples were positive for cowpea mild mottle virus (CPMMV). Among the common bean genotypes screened for resistance to CPMMV, only RO/2/1 and Giza 3 were resistant to the disease.(Author)
A laser ablation ICP-MS based method for multiplexed immunoblot analysis
DEFF Research Database (Denmark)
de Bang, Thomas Christian; Petersen, Jørgen; Pedas, Pai Rosager
2015-01-01
developed a multiplexed antibody-based assay and analysed selected PSII subunits in barley (Hordeum vulgare L.). A selection of antibodies were labelled with specific lanthanides and immunoreacted with thylakoids exposed to Mn deficiency after western blotting. Subsequently, western blot membranes were...... analysed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), which allowed selective and relative quantitative analysis via the different lanthanides. The method was evaluated against established liquid chromatography electrospray ionization tandem mass spectrometry (LC...... by more than one technique. The developed method enables a higher number of proteins to be multiplexed in comparison to existing immunoassays. Furthermore, multiplexed protein analysis by LA-ICP-MS provides an analytical platform with high throughput appropriate for screening large collections of plants....
Imaging Techniques in Endodontics: An Overview
Deepak, B. S.; Subash, T. S.; Narmatha, V. J.; Anamika, T.; Snehil, T. K.; Nandini, D. B.
2012-01-01
This review provides an overview of the relevance of imaging techniques such as, computed tomography, cone beam computed tomography, and ultrasound, to endodontic practice. Many limitations of the conventional radiographic techniques have been overcome by the newer methods. Advantages and disadvantages of various imaging techniques in endodontic practice are also discussed. PMID:22530184
Motion Transplantation Techniques: A Survey
van Basten, Ben; Egges, Arjan
2012-01-01
During the past decade, researchers have developed several techniques for transplanting motions. These techniques transplant a partial auxiliary motion, possibly defined for a small set of degrees of freedom, on a base motion. Motion transplantation improves motion databases' expressiveness and
Maple, Peter A C; Beard, Stuart; Parry, Ruth P; Brown, Kevin E
2013-10-01
Human parvovirus 4 (ParV4), a newly described member of the family Parvoviridae, like B19V, has been found in pooled plasma preparations. The extent, and significance, of ParV4 exposure in UK blood donors remain to be determined and reliable detection of ParV4 immunoglobulin (Ig)G, using validated methods, is needed. With ParV4 virus-like particles a ParV4 IgG time-resolved fluorescence immunoassay (TRFIA) was developed. There is no gold standard or reference assay for measuring ParV4 IgG and the utility of the TRFIA was first examined using a panel of sera from people who inject drugs (PWIDS)--a high-prevalence population for ParV4 infection. Western blotting was used to confirm the specificity of TRFIA-reactive sera. Two cohorts of UK blood donor sera comprising 452 sera collected in 1999 and 156 sera collected in 2009 were tested for ParV4 IgG. Additional testing for B19V IgG, hepatitis C virus antibodies (anti-HCV), and ParV4 DNA was also undertaken. The rate of ParV4 IgG seroprevalence in PWIDS was 20.7% and ParV4 IgG was positively associated with the presence of anti-HCV with 68.4% ParV4 IgG-positive sera testing anti-HCV-positive versus 17.1% ParV4 IgG-negative sera. Overall seropositivity for ParV4 IgG, in 608 UK blood donors was 4.76%. The ParV4 IgG seropositivity for sera collected in 1999 was 5.08%, compared to 3.84% for sera collected in 2009. No ParV4 IgG-positive blood donor sera had detectable ParV4 DNA. ParV4 IgG has been found in UK blood donors and this finding needs further investigation. © 2013 American Association of Blood Banks.
Xiao, Meng; Kong, Fanrong; Sorrell, Tania C; Cao, Yongyan; Lee, Ok Cha; Liu, Ying; Sintchenko, Vitali; Chen, Sharon C A
2010-02-01
Although 16S rRNA gene sequence analysis is employed most often for the definitive identification of Nocardia species, alternate molecular methods and polymorphisms in other gene targets have also enabled species determinations. We evaluated a combined Nocardia PCR-based reverse line blot (RLB) hybridization assay based on 16S and 16S-23S rRNA gene spacer region polymorphisms to identify 12 American Type Culture Collection and 123 clinical Nocardia isolates representing 14 species; results were compared with results from 16S rRNA gene sequencing. Thirteen 16S rRNA gene-based (two group-specific and 11 species-specific) and five 16S-23S spacer-targeted (two taxon-specific and three species-specific) probes were utilized. 16S rRNA gene-based probes correctly identified 124 of 135 isolates (sensitivity, 92%) but were unable to identify Nocardia paucivorans strains (n = 10 strains) and a Nocardia asteroides isolate with a novel 16S rRNA gene sequence. Nocardia farcinica and Nocardia cyriacigeorgica strains were identified by the sequential use of an N. farcinica-"negative" probe and a combined N. farcinica/N. cyriacigeorgica probe. The assay specificity was high (99%) except for weak cross-reactivity between the Nocardia brasiliensis probe with the Nocardia thailandica DNA product; however, cross-hybridization with closely related nontarget species may occur. The incorporation of 16S-23S rRNA gene spacer-based probes enabled the identification of all N. paucivorans strains. The overall sensitivity using both probe sets was >99%. Both N. farcinica-specific 16S-23S rRNA gene spacer-directed probes were required to identify all N. farcinica stains by using this probe set. The study demonstrates the utility of a combined PCR/RLB assay for the identification of clinically relevant Nocardia species and its potential for studying subtypes of N. farcinica. Where species assignment is ambiguous or not possible, 16S rRNA gene sequencing is recommended.
Tendon 'turnover lengthening' technique.
Cerovac, S; Miranda, B H
2013-11-01
Tendon defect reconstruction is amongst the most technically challenging areas in hand surgery. Tendon substance deficiency reconstruction techniques include lengthening, grafting, two-stage reconstruction and tendon transfers, however each is associated with unique challenges over and above direct repair. We describe a novel 'turnover lengthening' technique for hand tendons that has successfully been applied to the repair of several cases, including a case of attritional flexor and traumatic extensor tendon rupture in two presented patients where primary tenorrhaphy was not possible. In both cases a good post-operative outcome was achieved, as the patients were happy having returned back to normal activities of daily living such that they were discharged 12 weeks post-operatively. Our technique avoids the additional morbidity and complications associated with grafting, transfers and two stage reconstructions. It is quick, simple and reproducible for defects not exceeding 3-4 cm, provides a means of immediate one stage reconstruction, no secondary donor site morbidity and does not compromise salvage by tendon transfer and/or two-stage reconstruction in cases of failure. To our knowledge no such technique has been previously been described to reconstruct such hand tendon defects. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.
The attribute measurement technique
International Nuclear Information System (INIS)
MacArthur, Duncan W.; Langner, Diana; Smith, Morag; Thron, Jonathan; Razinkov, Sergey; Livke, Alexander
2010-01-01
Any verification measurement performed on potentially classified nuclear material must satisfy two seemingly contradictory constraints. First and foremost, no classified information can be released. At the same time, the monitoring party must have confidence in the veracity of the measurement. An information barrier (IB) is included in the measurement system to protect the potentially classified information while allowing sufficient information transfer to occur for the monitoring party to gain confidence that the material being measured is consistent with the host's declarations, concerning that material. The attribute measurement technique incorporates an IB and addresses both concerns by measuring several attributes of the nuclear material and displaying unclassified results through green (indicating that the material does possess the specified attribute) and red (indicating that the material does not possess the specified attribute) lights. The attribute measurement technique has been implemented in the AVNG, an attribute measuring system described in other presentations at this conference. In this presentation, we will discuss four techniques used in the AVNG: (1) the 1B, (2) the attribute measurement technique, (3) the use of open and secure modes to increase confidence in the displayed results, and (4) the joint design as a method for addressing both host and monitor needs.
Practical techniques for pediatric computed tomography
International Nuclear Information System (INIS)
Fitz, C.R.; Harwood-Nash, D.C.; Kirks, D.R.; Kaufman, R.A.; Berger, P.E.; Kuhn, J.P.; Siegel, M.J.
1983-01-01
Dr. Donald Kirks has assembled this section on Practical Techniques for Pediatric Computed Tomography. The material is based on a presentation in the Special Interest session at the 25th Annual Meeting of the Society for Pediatric Radiology in New Orleans, Louisiana, USA in 1982. Meticulous attention to detail and technique is required to ensure an optimal CT examination. CT techniques specifically applicable to infants and children have not been disseminated in the radiology literature and in this respect it may rightly be observed that ''the child is not a small adult''. What follows is a ''cookbook'' prepared by seven participants and it is printed in Pediatric Radiology, in outline form, as a statement of individual preferences for pediatric CT techniques. This outline gives concise explanation of techniques and permits prompt dissemination of information. (orig.)
Advanced sensing techniques for cognitive radio
Zhao, Guodong; Li, Shaoqian
2017-01-01
This SpringerBrief investigates advanced sensing techniques to detect and estimate the primary receiver for cognitive radio systems. Along with a comprehensive overview of existing spectrum sensing techniques, this brief focuses on the design of new signal processing techniques, including the region-based sensing, jamming-based probing, and relay-based probing. The proposed sensing techniques aim to detect the nearby primary receiver and estimate the cross-channel gain between the cognitive transmitter and primary receiver. The performance of the proposed algorithms is evaluated by simulations in terms of several performance parameters, including detection probability, interference probability, and estimation error. The results show that the proposed sensing techniques can effectively sense the primary receiver and improve the cognitive transmission throughput. Researchers and postgraduate students in electrical engineering will find this an exceptional resource.
The Technique of Special-Effects Cinematography.
Fielding, Raymond
The author describes the many techniques used to produce cinematic effects that would be too costly, too difficult, too time-consuming, too dangerous, or simply impossible to achieve with conventional photographic techniques. He points out that these techniques are available not only for 35 millimeter work but also to the 16 mm. photographer who…
International Nuclear Information System (INIS)
Charlton, J.S.
1986-01-01
The way in which neutrons interact with matter such as slowing-down, diffusion, neutron absorption and moderation are described. The use of neutron techniques in industry, in moisture gages, level and interface measurements, the detection of blockages, boron analysis in ore feedstock and industrial radiography are discussed. (author)
Royuela, M; García-Anchuelo, R; Arenas, M I; Cervera, M; Fraile, B; Paniagua, R
1996-04-01
The presence and distribution pattern of paramyosin have been examined in different invertebrate muscle cell types by means of Western blot analysis and electron microscopy immunogold labelling. The muscles studied were: transversely striated muscle with continuous Z lines (flight muscle from Drosophila melanogaster), transversely striated muscle with discontinuous Z lines (heart muscle from the snail Helix aspersa), obliquely striated body wall muscle from the earthworm Eisenia foetida, and smooth muscles (retractor muscle from the snail and pseudoheart outer muscular layer from the earthworm). Paramyosin-like immunoreactivity was localized in thick filaments of all muscles studied. Immunogold particle density was similar along the whole thick filament length in insect flight muscle but it predominated in filament tips of fusiform thick filaments in both snail heart and earthworm body wall musculature when these filaments were observed in longitudinal sections. In obliquely sectioned thick filaments, immunolabelling was more abundant at the sites where filaments disappeared from the section. These results agree with the notion that paramyosin extended along the whole filament length, but that it can only be immunolabelled when it is not covered by myosin. In all muscles examined, immunolabelling density was lower in cross-sectioned myofilaments than in longitudinally sectioned myofilaments. This suggests that paramyosin does not form a continuous filament. The results of a semiquantitative analysis of paramyosin-like immunoreactivity indicated that it was more abundant in striated than in smooth muscles, and that, within striated muscles, transversely striated muscles contain more paramyosin than obliquely striated muscles.
Applying contemporary statistical techniques
Wilcox, Rand R
2003-01-01
Applying Contemporary Statistical Techniques explains why traditional statistical methods are often inadequate or outdated when applied to modern problems. Wilcox demonstrates how new and more powerful techniques address these problems far more effectively, making these modern robust methods understandable, practical, and easily accessible.* Assumes no previous training in statistics * Explains how and why modern statistical methods provide more accurate results than conventional methods* Covers the latest developments on multiple comparisons * Includes recent advanc
Materials characterization techniques
National Research Council Canada - National Science Library
Zhang, Sam; Li, L; Kumar, Ashok
2009-01-01
"With an emphasis on practical applications and real-world case studies, Materials Characterization Techniques presents the principles of widely used advanced surface and structural characterization...
A disposition of interpolation techniques
Knotters, M.; Heuvelink, G.B.M.
2010-01-01
A large collection of interpolation techniques is available for application in environmental research. To help environmental scientists in choosing an appropriate technique a disposition is made, based on 1) applicability in space, time and space-time, 2) quantification of accuracy of interpolated
Reddy, S Srikanth; Revathi, Kakkirala; Reddy, S Kranthikumar
2013-01-01
Conventional casting technique is time consuming when compared to accelerated casting technique. In this study, marginal accuracy of castings fabricated using accelerated and conventional casting technique was compared. 20 wax patterns were fabricated and the marginal discrepancy between the die and patterns were measured using Optical stereomicroscope. Ten wax patterns were used for Conventional casting and the rest for Accelerated casting. A Nickel-Chromium alloy was used for the casting. The castings were measured for marginal discrepancies and compared. Castings fabricated using Conventional casting technique showed less vertical marginal discrepancy than the castings fabricated by Accelerated casting technique. The values were statistically highly significant. Conventional casting technique produced better marginal accuracy when compared to Accelerated casting. The vertical marginal discrepancy produced by the Accelerated casting technique was well within the maximum clinical tolerance limits. Accelerated casting technique can be used to save lab time to fabricate clinical crowns with acceptable vertical marginal discrepancy.
Radiotracer techniques in hydrological studies
International Nuclear Information System (INIS)
Oladipo, M.O.A.; Funtua, I.I.
2000-07-01
The use of radioactive tracers particularly short-lived radioisotopes frequently offers advantages over conventional methods of analyses. Applications of nuclear techniques in the field of hydrology constitute important and sometimes unique tools for obtaining critical information needed for water resources management. Essentially, radiotracer techniques offer a safe, cost effective and powerful tool in the assessment, management and protection of water resources. The Centre for Energy Research and Training, Ahmadu Bello University, Zaria of late has been offering consultancy services to some industries in the area of radiotracer technique. The first nuclear reactor in Nigeria, the MNSR, is expected to be commissioned in the Centre very soon. Many short-lived radioisotopes such as Cu-64, Ga-72, Br-82, Hg-197 etc which are very important in hydrological studies can be produced by the MNSR facility. This article reports on the basic principles of the technique and its roles in hydrology
International Nuclear Information System (INIS)
Ogawa, Ryuichiro; Ishijima, Noboru
1999-02-01
Decommissioning techniques such as radiation measuring and monitoring, decontamination, dismantling and remote handling in the world were surveyed to upgrading technical know-how database for decommissioning of Joyo Waste Treatment Facility (JWTF). As the result, five literatures for measuring and monitoring techniques, 14 for decontamination and 22 for dismantling feasible for JWTF decommissioning were obtained and were summarized in tables. On the basis of the research, practical applicability of those techniques to decommissioning of JWTF was evaluated. This report contains brief surveyed summaries related to JWTF decommissioning. (H. Itami)
Weighted hybrid technique for recommender system
Suriati, S.; Dwiastuti, Meisyarah; Tulus, T.
2017-12-01
Recommender system becomes very popular and has important role in an information system or webpages nowadays. A recommender system tries to make a prediction of which item a user may like based on his activity on the system. There are some familiar techniques to build a recommender system, such as content-based filtering and collaborative filtering. Content-based filtering does not involve opinions from human to make the prediction, while collaborative filtering does, so collaborative filtering can predict more accurately. However, collaborative filtering cannot give prediction to items which have never been rated by any user. In order to cover the drawbacks of each approach with the advantages of other approach, both approaches can be combined with an approach known as hybrid technique. Hybrid technique used in this work is weighted technique in which the prediction score is combination linear of scores gained by techniques that are combined.The purpose of this work is to show how an approach of weighted hybrid technique combining content-based filtering and item-based collaborative filtering can work in a movie recommender system and to show the performance comparison when both approachare combined and when each approach works alone. There are three experiments done in this work, combining both techniques with different parameters. The result shows that the weighted hybrid technique that is done in this work does not really boost the performance up, but it helps to give prediction score for unrated movies that are impossible to be recommended by only using collaborative filtering.
Twin - Arch technique. Revival of the "edgewise -Technique"?
Directory of Open Access Journals (Sweden)
Jacob Karp
2012-01-01
The SNB – Bracket brings a new dimension into the orthodontic world which is most apparent in extraction cases. Its Teflon – like material has a very low friction coefficient thus, reducing the treatment time considerably. Through the use of low dimensioned arch wires, the Twin – Arch Technique becomes a Light – wire system and simultaneously provides good anchorage and torque control.
Spray deposition using impulse atomization technique
International Nuclear Information System (INIS)
Ellendt, N.; Schmidt, R.; Knabe, J.; Henein, H.; Uhlenwinkel, V.
2004-01-01
A novel technique, impulse atomization, has been used for spray deposition. This single fluid atomization technique leads to different spray characteristics and impact conditions of the droplets compared to gas atomization technique which is the common technique used for spray deposition. Deposition experiments with a Cu-6Sn alloy were conducted to evaluate the appropriateness of impulse atomization to produce dense material. Based on these experiments, a model has been developed to simulate the thermal history and the local solidification rates of the deposited material. A numerical study shows how different cooling conditions affect the solidification rate of the material
Invasive and noninvasive dental analgesia techniques.
Estafan, D J
1998-01-01
Although needle-administered local anesthesia has been an essential tool of modern dentistry, it has also been responsible for many patients' fears of dental visits. Several new techniques have recently evolved that may offer viable alternatives. Two of these operate via electronic mechanisms that interfere with pain signals, two others involve transmucosal modes of administration, and a fifth technique involves an intraosseous pathway for anesthesia administration. Each of these techniques has different indications for dental procedures, but none is intended to replace needle administration in dentistry. This overview highlights the salient features of these alternative dental anesthesia techniques.
Influence of DNA treatments on Southern blot hybridization analysis ...
African Journals Online (AJOL)
STORAGESEVER
2008-06-03
Jun 3, 2008 ... DNA samples obtained by a non-phenol/chloroform isolation method, from three races of Fusarium oxysporum f. sp. lycopersici ... Key words: Fusarium oxysporum, DIG-IGS Probe, Southern hybridization. INTRODUCTION .... Detection of Fusarium spp in plants with monoclonal antibody. Ann. Phytopathol.
Letbanen er blot et led i mobilitetskæden
DEFF Research Database (Denmark)
Olesen, Mette
2014-01-01
Letbaner bliver ikke en succes alene; de skal ses i helhed med de øvrige transportmidler og tiltag, der fremmer deres potentiale. Artiklen retter fokus mod netop nogle af de mange mobilitetsløsninger, der i en europæisk sammenhæng har fremmet letbanens potentiale – løsninger, der kan være relevan...
Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots
Directory of Open Access Journals (Sweden)
Sanne M. M. Hensen
2015-04-01
Full Text Available Citrullination is the conversion of peptidylarginine to peptidylcitrulline, which is catalyzed by peptidylarginine deiminases. This conversion is involved in different physiological processes and is associated with several diseases, including cancer and rheumatoid arthritis. A common method to detect citrullinated proteins relies on anti-modified citrulline antibodies directed to a specific chemical modification of the citrulline side chain. Here, we describe a versatile, antibody-independent method for the detection of citrullinated proteins on a membrane, based on the selective reaction of phenylglyoxal with the ureido group of citrulline under highly acidic conditions. The method makes use of 4-azidophenylglyoxal, which, after reaction with citrullinated proteins, can be visualized with alkyne-conjugated probes. The sensitivity of this procedure, using an alkyne-biotin probe, appeared to be comparable to the antibody-based detection method and independent of the sequence surrounding the citrulline.
Physical simulations using centrifuge techniques
International Nuclear Information System (INIS)
Sutherland, H.J.
1981-01-01
Centrifuge techniques offer a technique for doing physical simulations of the long-term mechanical response of deep ocean sediment to the emplacement of waste canisters and to the temperature gradients generated by them. Preliminary investigations of the scaling laws for pertinent phenomena indicate that the time scaling will be consistent among them and equal to the scaling factor squared. This result implies that this technique will permit accelerated-life-testing of proposed configurations; i.e, long-term studies may be done in relatively short times. Presently, existing centrifuges are being modified to permit scale model testing. This testing will start next year
LANDING TECHNIQUES IN BEACH VOLLEYBALL
Directory of Open Access Journals (Sweden)
Markus Tilp
2013-09-01
Full Text Available The aims of the present study were to establish a detailed and representative record of landing techniques (two-, left-, and right-footed landings in professional beach volleyball and compare the data with those of indoor volleyball. Beach volleyball data was retrieved from videos taken at FIVB World Tour tournaments. Landing techniques were compared in the different beach and indoor volleyball skills serve, set, attack, and block with regard to sex, playing technique, and court position. Significant differences were observed between men and women in landings following block actions (χ²(2 = 18.19, p < 0.01 but not following serve, set, and attack actions. Following blocking, men landed more often on one foot than women. Further differences in landings following serve and attack with regard to playing technique and position were mainly observed in men. The comparison with landing techniques in indoor volleyball revealed overall differences both in men (χ²(2 = 161.4, p < 0.01 and women (χ²(2 = 84.91, p < 0.01. Beach volleyball players land more often on both feet than indoor volleyball players. Besides the softer surface in beach volleyball, and therefore resulting lower loads, these results might be another reason for fewer injuries and overuse conditions compared to indoor volleyball
Landing Techniques in Beach Volleyball
Tilp, Markus; Rindler, Michael
2013-01-01
The aims of the present study were to establish a detailed and representative record of landing techniques (two-, left-, and right-footed landings) in professional beach volleyball and compare the data with those of indoor volleyball. Beach volleyball data was retrieved from videos taken at FIVB World Tour tournaments. Landing techniques were compared in the different beach and indoor volleyball skills serve, set, attack, and block with regard to sex, playing technique, and court position. Significant differences were observed between men and women in landings following block actions (χ2(2) = 18.19, p volleyball revealed overall differences both in men (χ2(2) = 161.4, p volleyball players land more often on both feet than indoor volleyball players. Besides the softer surface in beach volleyball, and therefore resulting lower loads, these results might be another reason for fewer injuries and overuse conditions compared to indoor volleyball. Key Points About 1/3 of all jumping actions in beach volleyball result in a landing on one foot. Especially following block situations men land on one foot more often than women. Landing techniques are related to different techniques and positions. Landings on one foot are less common in beach volleyball than indoor volleyball. This could be a reason for fewer injuries and overuse conditions. PMID:24149150
Lu, Ying-Ying; Chen, Tong-Sheng; Wang, Xiao-Ping; Li, Li
2010-07-01
Dihydroartemisinin (DHA), a front-line antimalarial herbal compound, has been shown to possess promising anticancer activity with low toxicity. We have previously reported that DHA induced caspase-3-dependent apoptosis in human lung adenocarcinoma cells. However, the cellular target and molecular mechanism of DHA-induced apoptosis is still poorly defined. We use confocal fluorescence microscopy imaging, fluorescence resonance energy transfer, and fluorescence recovery after photobleaching techniques to explore the roles of DHA-elicited reactive oxygen species (ROS) in the DHA-induced Bcl-2 family proteins activation, mitochondrial dysfunction, caspase cascade, and cell death. Cell Counting Kit-8 assay and flow cytometry analysis showed that DHA induced ROS-mediated apoptosis. Confocal imaging analysis in a single living cell and Western blot assay showed that DHA triggered ROS-dependent Bax translocation, mitochondrial membrane depolarization, alteration of mitochondrial morphology, cytochrome c release, caspase-9, caspase-8, and caspase-3 activation, indicating the coexistence of ROS-mediated mitochondrial and death receptor pathway. Collectively, our findings demonstrate for the first time that DHA induces cell apoptosis by triggering ROS-mediated caspase-8/Bid activation and the mitochondrial pathway, which provides some novel insights into the application of DHA as a potential anticancer drug and a new therapeutic strategy by targeting ROS signaling in lung adenocarcinoma therapy in the future.
Song, Juquan; Zhang, Xiao-jun; Boehning, Darren; Brooks, Natasha C.; Herndon, David N.; Jeschke, Marc G.
2012-01-01
Severe burn-induced liver damage and dysfunction is associated with endoplasmic reticulum (ER) stress. ER stress has been shown to regulate global protein synthesis. In the current study, we induced ER stress in vitro and estimated the effect of ER stress on hepatic protein synthesis. The aim was two-fold: (1) to establish an in vitro model to isotopically measure hepatic protein synthesis and (2) to evaluate protein fractional synthetic rate (FSR) in response to ER stress. Human hepatocellular carcinoma cells (HepG2) were cultured in medium supplemented with stable isotopes 1,2-13C2-glycine and L-[ring-13C6]phenylalanine. ER stress was induced by exposing the cells to 100 nM of thapsigargin (TG). Cell content was collected from day 0 to 14. Alterations in cytosolic calcium were measured by calcium imaging and ER stress markers were confirmed by Western blotting. The precursor and product enrichments were detected by GC-MS analysis for FSR calculation. We found that the hepatic protein FSR were 0.97±0.02 and 0.99±0.05%/hr calculated from 1,2-13C2-glycine and L-[ring-13C6]phenylalanine, respectively. TG depleted ER calcium stores and induced ER stress by upregulating p-IRE-1 and Bip. FSR dramatically decreased to 0.68±0.03 and 0.60±0.06%/hr in the TG treatment group (pisotope tracer incorporation technique is a useful method for studying the effects of ER stress on hepatic protein synthesis. PMID:22298954
Biopsy techniques for intraocular tumors
Directory of Open Access Journals (Sweden)
Pukhraj Rishi
2016-01-01
Full Text Available Biopsy involves the surgical removal of a tissue specimen for histopathologic evaluation. Most intraocular tumors are reliably diagnosed based on the clinical evaluation or with noninvasive diagnostic techniques. However, accurately diagnosing a small percentage of tumors can be challenging. A tissue biopsy is thus needed to establish a definitive diagnosis and plan the requisite treatment. From fine-needle aspiration biopsy (FNAB to surgical excision, all tissue collection techniques have been studied in the literature. Each technique has its indications and limitations. FNAB has been reported to provide for 88-95% reliable and safe ophthalmic tumor diagnosis and has gained popularity for prognostic purposes and providing eye conserving treatment surgeries. The technique and instrumentation for biopsy vary depending upon the tissue involved (retina, choroid, subretinal space, vitreous, and aqueous, suspected diagnosis, size, location, associated retinal detachment, and clarity of the media. The cytopathologist confers a very important role in diagnosis and their assistance plays a key role in managing and planning the treatment for malignancies.
Submucosal tunneling techniques: current perspectives.
Kobara, Hideki; Mori, Hirohito; Rafiq, Kazi; Fujihara, Shintaro; Nishiyama, Noriko; Ayaki, Maki; Yachida, Tatsuo; Matsunaga, Tae; Tani, Johji; Miyoshi, Hisaaki; Yoneyama, Hirohito; Morishita, Asahiro; Oryu, Makoto; Iwama, Hisakazu; Masaki, Tsutomu
2014-01-01
Advances in endoscopic submucosal dissection include a submucosal tunneling technique, involving the introduction of tunnels into the submucosa. These tunnels permit safer offset entry into the peritoneal cavity for natural orifice transluminal endoscopic surgery. Technical advantages include the visual identification of the layers of the gut, blood vessels, and subepithelial tumors. The creation of a mucosal flap that minimizes air and fluid leakage into the extraluminal cavity can enhance the safety and efficacy of surgery. This submucosal tunneling technique was adapted for esophageal myotomy, culminating in its application to patients with achalasia. This method, known as per oral endoscopic myotomy, has opened up the new discipline of submucosal endoscopic surgery. Other clinical applications of the submucosal tunneling technique include its use in the removal of gastrointestinal subepithelial tumors and endomicroscopy for the diagnosis of functional and motility disorders. This review suggests that the submucosal tunneling technique, involving a mucosal safety flap, can have potential values for future endoscopic developments.
Lightweight Cryptographic Techniques
National Research Council Canada - National Science Library
Yuen, Horace
2004-01-01
The objective of this project was to develop new cryptographic techniques, and to modify the important existing ones, for applications to encryption and authentication in energy-constrained sensors...
Energy Technology Data Exchange (ETDEWEB)
Anon.
1998-09-01
The ``hydro-wired`` technique consists in the use of small diameter synthetic pipes for the water supply of hot or cold water to space heating or cooling appliances. This new technique has several advantages regarding the thermal comfort and the realization of installations. However cares must be taken during the application of this technique and the design of installations. This paper describes: the French potential market for this technique (heating and cooling floors, traditional heating, sanitary hot water distribution), the characteristics of this technique with respect to traditional techniques (the sheathing of pipes, the specificities of the `octopus`-type distribution), and the design of installations (the different steps: preliminary thermal analysis, realization, start-up and tests). (J.S.)
THE TECHNIQUES IN TEACHING LISTENING SKILL
Directory of Open Access Journals (Sweden)
Hidayah Nor
2015-12-01
Full Text Available Listening is very important skill in language because by listening students can produce language like speaking and writing by vocabulary that they get from listening. The English teacher of MAN 3 Banjarmasin used some techniques in teaching listening using the facilities in language laboratory such as tape cassette, television, and VCD/DVD. This research described the techniques in teaching listening skill of the Islamic high school students. The subjects of this study were an English teacher and 48 students of the tenth grade at MAN 3 Banjarmasin in Academic Year 2009/2010. To collect the data, it was used some techniques such as observation, interview, and documentary. Then all data were analyzed using descriptive method qualitatively and quantitatively, by concluding inductively. The result indicates that the techniques in teaching listening applied by the English teacher of the tenth grade students at MAN 3 Banjarmasin in Academic Year 2009/2010 are: Information Transfer, Paraphrasing and Translating, Answering Questions, Summarizing, Filling in Blanks, and Answering to Show Comprehension of Messages. The students’ ability of listening comprehension using six techniques is categorized in very high, high, and average levels. Keywords: listening techniques, teaching listening skill
Intraosseous anesthesia: implications, instrumentation and techniques.
Kleber, Christopher H
2003-04-01
The author reviews historical methods and the instruments used to bring about intraosseous anesthesia, or IOA; discusses the criteria for successful use of the intraosseous injection, or IOI, technique; and provides recommendations. Articles from before 1990 consisted of subjective reports of patient types and procedures performed using IOI as a primary technique. Studies published after 1990 yielded subjective findings on indications for expanded clinical use. The author discusses the expansion of the role of IOI relative to integrated local anesthetic delivery systems. The literature and studies verify the efficacy of IOI as a supplemental or primary technique. The author recommends anesthetics and infusion sites, and reports on the patients' perceptions of comfort. IOI can be used as a supplemental or primary technique to bring about local anesthesia in routine dental procedures. It can be used as a supplemental technique with mandibular nerve blocks to enhance deep pulpal anesthesia. It can be used as a primary technique so that patients do not experience numb lips or tongues postoperatively. Dentists can appreciate the immediate onset of anesthesia and reduced dosage levels of anesthetics associated with using IOI.
A video authentication technique
International Nuclear Information System (INIS)
Johnson, C.S.
1987-01-01
Unattended video surveillance systems are particularly vulnerable to the substitution of false video images into the cable that connects the camera to the video recorder. New technology has made it practical to insert a solid state video memory into the video cable, freeze a video image from the camera, and hold this image as long as desired. Various techniques, such as line supervision and sync detection, have been used to detect video cable tampering. The video authentication technique described in this paper uses the actual video image from the camera as the basis for detecting any image substitution made during the transmission of the video image to the recorder. The technique, designed for unattended video systems, can be used for any video transmission system where a two-way digital data link can be established. The technique uses similar microprocessor circuitry at the video camera and at the video recorder to select sample points in the video image for comparison. The gray scale value of these points is compared at the recorder controller and if the values agree within limits, the image is authenticated. If a significantly different image was substituted, the comparison would fail at a number of points and the video image would not be authenticated. The video authentication system can run as a stand-alone system or at the request of another system
Acceleration techniques for the discrete ordinate method
International Nuclear Information System (INIS)
Efremenko, Dmitry; Doicu, Adrian; Loyola, Diego; Trautmann, Thomas
2013-01-01
In this paper we analyze several acceleration techniques for the discrete ordinate method with matrix exponential and the small-angle modification of the radiative transfer equation. These techniques include the left eigenvectors matrix approach for computing the inverse of the right eigenvectors matrix, the telescoping technique, and the method of false discrete ordinate. The numerical simulations have shown that on average, the relative speedup of the left eigenvector matrix approach and the telescoping technique are of about 15% and 30%, respectively. -- Highlights: ► We presented the left eigenvector matrix approach. ► We analyzed the method of false discrete ordinate. ► The telescoping technique is applied for matrix operator method. ► Considered techniques accelerate the computations by 20% in average.
Diagram Techniques in Group Theory
Stedman, Geoffrey E.
2009-09-01
Preface; 1. Elementary examples; 2. Angular momentum coupling diagram techniques; 3. Extension to compact simple phase groups; 4. Symmetric and unitary groups; 5. Lie groups and Lie algebras; 6. Polarisation dependence of multiphoton processes; 7. Quantum field theoretic diagram techniques for atomic systems; 8. Applications; Appendix; References; Indexes.
Constant exposure technique in industrial radiography
International Nuclear Information System (INIS)
Domanus, J.C.
1983-08-01
The principles and advantages of the constant exposure technique are explained. Choice of exposure factors is analyzed. Film, paper and intensifying screens used throughout the investigation and film and paper processing are described. Exposure technique and the use of image quality indicators are given. Methods of determining of radiographic image quality are presented. Conclusions about the use of constant exposure vs. constant kilovoltage technique are formulated. (author)
Huffman-based code compression techniques for embedded processors
Bonny, Mohamed Talal
2010-09-01
The size of embedded software is increasing at a rapid pace. It is often challenging and time consuming to fit an amount of required software functionality within a given hardware resource budget. Code compression is a means to alleviate the problem by providing substantial savings in terms of code size. In this article we introduce a novel and efficient hardware-supported compression technique that is based on Huffman Coding. Our technique reduces the size of the generated decoding table, which takes a large portion of the memory. It combines our previous techniques, Instruction Splitting Technique and Instruction Re-encoding Technique into new one called Combined Compression Technique to improve the final compression ratio by taking advantage of both previous techniques. The instruction Splitting Technique is instruction set architecture (ISA)-independent. It splits the instructions into portions of varying size (called patterns) before Huffman coding is applied. This technique improves the final compression ratio by more than 20% compared to other known schemes based on Huffman Coding. The average compression ratios achieved using this technique are 48% and 50% for ARM and MIPS, respectively. The Instruction Re-encoding Technique is ISA-dependent. It investigates the benefits of reencoding unused bits (we call them reencodable bits) in the instruction format for a specific application to improve the compression ratio. Reencoding those bits can reduce the size of decoding tables by up to 40%. Using this technique, we improve the final compression ratios in comparison to the first technique to 46% and 45% for ARM and MIPS, respectively (including all overhead that incurs). The Combined Compression Technique improves the compression ratio to 45% and 42% for ARM and MIPS, respectively. In our compression technique, we have conducted evaluations using a representative set of applications and we have applied each technique to two major embedded processor architectures
Neutron techniques in Safeguards
International Nuclear Information System (INIS)
Zucker, M.S.
1982-01-01
An essential part of Safeguards is the ability to quantitatively and nondestructively assay those materials with special neutron-interactive properties involved in nuclear industrial or military technology. Neutron techniques have furnished most of the important ways of assaying such materials, which is no surprise since the neutronic properties are what characterizes them. The techniques employed rely on a wide selection of the many methods of neutron generation, detection, and data analysis that have been developed for neutron physics and nuclear science in general
Radiographic constant exposure technique
DEFF Research Database (Denmark)
Domanus, Joseph Czeslaw
1985-01-01
The constant exposure technique has been applied to assess various industrial radiographic systems. Different X-ray films and radiographic papers of two producers were compared. Special attention was given to fast film and paper used with fluorometallic screens. Radiographic image quality...... was tested by the use of ISO wire IQI's and ASTM penetrameters used on Al and Fe test plates. Relative speed and reduction of kilovoltage obtained with the constant exposure technique were calculated. The advantages of fast radiographic systems are pointed out...
Neyton, Lionel; Barth, Johannes; Nourissat, Geoffroy; Métais, Pierre; Boileau, Pascal; Walch, Gilles; Lafosse, Laurent
2018-05-19
To analyze graft and fixation (screw and EndoButton) positioning after the arthroscopic Latarjet technique with 2-dimensional computed tomography (CT) and to compare it with the open technique. We performed a retrospective multicenter study (March 2013 to June 2014). The inclusion criteria included patients with recurrent anterior instability treated with the Latarjet procedure. The exclusion criterion was the absence of a postoperative CT scan. The positions of the hardware, the positions of the grafts in the axial and sagittal planes, and the dispersion of values (variability) were compared. The study included 208 patients (79 treated with open technique, 87 treated with arthroscopic Latarjet technique with screw fixation [arthro-screw], and 42 treated with arthroscopic Latarjet technique with EndoButton fixation [arthro-EndoButton]). The angulation of the screws was different in the open group versus the arthro-screw group (superior, 10.3° ± 0.7° vs 16.9° ± 1.0° [P open inferior screws (P = .003). In the axial plane (level of equator), the arthroscopic techniques resulted in lateral positions (arthro-screw, 1.5 ± 0.3 mm lateral [P open technique (0.9 ± 0.2 mm medial). At the level of 25% of the glenoid height, the arthroscopic techniques resulted in lateral positions (arthro-screw, 0.3 ± 0.3 mm lateral [P open technique (1.0 ± 0.2 mm medial). Higher variability was observed in the arthro-screw group. In the sagittal plane, the arthro-screw technique resulted in higher positions (55% ± 3% of graft below equator) and the arthro-EndoButton technique resulted in lower positions (82% ± 3%, P open technique (71% ± 2%). Variability was not different. This study shows that the position of the fixation devices and position of the bone graft with the arthroscopic techniques are statistically significantly different from those with the open technique with 2-dimensional CT assessment. In the sagittal plane, the arthro-screw technique provides the highest
Directory of Open Access Journals (Sweden)
Shalini Chandrasekhar
2018-01-01
Full Text Available Background: The aim of this study was to evaluate the efficiency of four different obturating techniques in filling the radicular space in primary teeth. Materials and Methods: This clinical trial was carried out on 34 healthy, cooperative children (5–9 years who had 63 carious primary teeth indicated for pulpectomy. They were divided into four groups, such that in each group, a total of 40 canals were allotted for obturation with respective technique. The root canals of selected primary teeth were filled with Endoflas obturating material using either bi-directional spiral (Group 1; incremental technique (Group 2, past inject (Group 3 or lentulo spiral (Group 4 according to the groups assigned. The effectiveness of the obturation techniques was assessed using postoperative radiographs. The assessment was made for a depth of fill in the canal, the presence of any voids using Modified Coll and Sadrian criteria. The obtained data were analyzed by using ANOVA test and unpaired t-test. Results: Bi-directional spiral and lentulo spiral were superior to other techniques in providing optimally filled canals (P< 0.05. The bi-directional spiral was superior to lentulo spiral in preventing overfill (P< 0.05. Conclusion: Based on the present study results, bi-directional spiral can be recommended as an alternate obturating technique in primary teeth.
Materials characterization techniques
National Research Council Canada - National Science Library
Zhang, Sam; Li, L; Kumar, Ashok
2009-01-01
... techniques for quality assurance, contamination control, and process improvement. The book reviews the most popular and powerful analysis and quality control tools, explaining the appropriate uses and related technical requirements...
Vibration transducer calibration techniques
Brinkley, D. J.
1980-09-01
Techniques for the calibration of vibration transducers used in the Aeronautical Quality Assurance Directorate of the British Ministry of Defence are presented. Following a review of the types of measurements necessary in the calibration of vibration transducers, the performance requirements of vibration transducers, which can be used to measure acceleration, velocity or vibration amplitude, are discussed, with particular attention given to the piezoelectric accelerometer. Techniques for the accurate measurement of sinusoidal vibration amplitude in reference-grade transducers are then considered, including the use of a position sensitive photocell and the use of a Michelson laser interferometer. Means of comparing the output of working-grade accelerometers with that of previously calibrated reference-grade devices are then outlined, with attention given to a method employing a capacitance bridge technique and a method to be used at temperatures between -50 and 200 C. Automatic calibration procedures developed to speed up the calibration process are outlined, and future possible extensions of system software are indicated.
Empirical techniques in finance
Bhar, Ramaprasad
2005-01-01
This book offers the opportunity to study and experience advanced empi- cal techniques in finance and in general financial economics. It is not only suitable for students with an interest in the field, it is also highly rec- mended for academic researchers as well as the researchers in the industry. The book focuses on the contemporary empirical techniques used in the analysis of financial markets and how these are implemented using actual market data. With an emphasis on Implementation, this book helps foc- ing on strategies for rigorously combing finance theory and modeling technology to extend extant considerations in the literature. The main aim of this book is to equip the readers with an array of tools and techniques that will allow them to explore financial market problems with a fresh perspective. In this sense it is not another volume in eco- metrics. Of course, the traditional econometric methods are still valid and important; the contents of this book will bring in other related modeling topics tha...
Radiation techniques for acromegaly
Directory of Open Access Journals (Sweden)
Minniti Giuseppe
2011-12-01
Full Text Available Abstract Radiotherapy (RT remains an effective treatment in patients with acromegaly refractory to medical and/or surgical interventions, with durable tumor control and biochemical remission; however, there are still concerns about delayed biochemical effect and potential late toxicity of radiation treatment, especially high rates of hypopituitarism. Stereotactic radiotherapy has been developed as a more accurate technique of irradiation with more precise tumour localization and consequently a reduction in the volume of normal tissue, particularly the brain, irradiated to high radiation doses. Radiation can be delivered in a single fraction by stereotactic radiosurgery (SRS or as fractionated stereotactic radiotherapy (FSRT in which smaller doses are delivered over 5-6 weeks in 25-30 treatments. A review of the recent literature suggests that pituitary irradiation is an effective treatment for acromegaly. Stereotactic techniques for GH-secreting pituitary tumors are discussed with the aim to define the efficacy and potential adverse effects of each of these techniques.
Radiation techniques for acromegaly
International Nuclear Information System (INIS)
Minniti, Giuseppe; Scaringi, Claudia; Enrici, Riccardo Maurizi
2011-01-01
Radiotherapy (RT) remains an effective treatment in patients with acromegaly refractory to medical and/or surgical interventions, with durable tumor control and biochemical remission; however, there are still concerns about delayed biochemical effect and potential late toxicity of radiation treatment, especially high rates of hypopituitarism. Stereotactic radiotherapy has been developed as a more accurate technique of irradiation with more precise tumour localization and consequently a reduction in the volume of normal tissue, particularly the brain, irradiated to high radiation doses. Radiation can be delivered in a single fraction by stereotactic radiosurgery (SRS) or as fractionated stereotactic radiotherapy (FSRT) in which smaller doses are delivered over 5-6 weeks in 25-30 treatments. A review of the recent literature suggests that pituitary irradiation is an effective treatment for acromegaly. Stereotactic techniques for GH-secreting pituitary tumors are discussed with the aim to define the efficacy and potential adverse effects of each of these techniques
Ion beam techniques in arts and archaeology
International Nuclear Information System (INIS)
Qin Guangyong; Pan Xianjia; Sun Zhongtian; Gao Zhengyao
1991-01-01
The ion beam techniques used in studies of arts and archaeology are compared with other analytical techniques. Some examples are specially selected to illustrate the achievements and trends of the techniques in this field
Radar rainfall image repair techniques
Directory of Open Access Journals (Sweden)
Stephen M. Wesson
2004-01-01
Full Text Available There are various quality problems associated with radar rainfall data viewed in images that include ground clutter, beam blocking and anomalous propagation, to name a few. To obtain the best rainfall estimate possible, techniques for removing ground clutter (non-meteorological echoes that influence radar data quality on 2-D radar rainfall image data sets are presented here. These techniques concentrate on repairing the images in both a computationally fast and accurate manner, and are nearest neighbour techniques of two sub-types: Individual Target and Border Tracing. The contaminated data is estimated through Kriging, considered the optimal technique for the spatial interpolation of Gaussian data, where the 'screening effect' that occurs with the Kriging weighting distribution around target points is exploited to ensure computational efficiency. Matrix rank reduction techniques in combination with Singular Value Decomposition (SVD are also suggested for finding an efficient solution to the Kriging Equations which can cope with near singular systems. Rainfall estimation at ground level from radar rainfall volume scan data is of interest and importance in earth bound applications such as hydrology and agriculture. As an extension of the above, Ordinary Kriging is applied to three-dimensional radar rainfall data to estimate rainfall rate at ground level. Keywords: ground clutter, data infilling, Ordinary Kriging, nearest neighbours, Singular Value Decomposition, border tracing, computation time, ground level rainfall estimation
Healthy Lifestyle Nutrition and healthy eating Healthy-cooking techniques capture the flavor and nutrients of food without extra fat or salt. By Mayo Clinic Staff Healthy cooking doesn't mean that ...
[Update in continuous renal replacement techniques].
Romero-García, M; de la Cueva-Ariza, L; Delgado-Hito, P
2013-01-01
Acute renal failure affects 25% of patients hospitalized in intensive care units. Despite technological advances, the mortality of these patients is still high due to its associated complications. Continuous renal replacement techniques are one of the treatments for acute renal failure because they make it possible to treat the complications and decrease mortality. The nurse's knowledge and skills regarding these techniques will be decisive for the success of the therapy. Consequently, the nurse's experience and training are key components. The objective of this article is to update the knowledge on continuous renal replacement techniques. Keeping this in mind, a review has been made of the physical and chemical principles such as diffusion and convection, among others. A description of the different continuous renal replacement techniques, a presentation of the main vascular access, and a description of the nursing cares and complications related to techniques used have also been provided. Copyright © 2012 Elsevier España, S.L. y SEEIUC. All rights reserved.
Kitchin, CR
2013-01-01
DetectorsOptical DetectionRadio and Microwave DetectionX-Ray and Gamma-Ray DetectionCosmic Ray DetectorsNeutrino DetectorsGravitational Radiation Dark Matter and Dark Energy Detection ImagingThe Inverse ProblemPhotographyElectronic ImagingScanningInterferometrySpeckle InterferometryOccultationsRadarElectronic ImagesPhotometryPhotometryPhotometersSpectroscopySpectroscopy SpectroscopesOther TechniquesAstrometryPolarimetrySolar StudiesMagnetometryComputers and The Internet.
Techniques for Improving Cash Management.
Lykins, Ronald G.
1973-01-01
This article deals with several techniques for regulating cash inflow and outflow and investing surplus cash for short periods of time. The techniques are: (1) consolidating checking accounts, (2) determining surplus cash by examining bank balances in conjunction with the cash book, (3) selecting a minimum bank balance, (4) investing a greater…
International Nuclear Information System (INIS)
1965-01-01
The medical and biological applications of radioisotopes depend on two basically different types of measurements, those on living subjects in vivo and those on samples in vitro. The International Atomic Energy Agency has in the past held several meetings on in vivo measurement techniques, notably whole-body counting and radioisotope scanning. The present volume contains the Proceedings of the first Symposium the Agency has organized to discuss the various aspects of techniques for sample measurement in vitro. The range of these sample measurement techniques is very wide. The sample may weigh a few milligrams or several hundred grams, and may be in the gaseous, liquid or solid state. Its radioactive content may consist of a single, known radioisotope or several unknown ones. The concentration of radioactivity may be low, medium or high. The measurements may be made manually or automatically and any one of the many radiation detectors now available may be used. The 53 papers presented at the Symposium illustrate the great variety of methods now in use for radioactive- sample measurements. The first topic discussed is gamma-ray spectrometry, which finds an increasing number of applications in sample measurements. Other sections of the Proceedings deal with: the use of computers in gamma-ray spectrometry and multiple tracer techniques; recent developments in activation analysis where both gamma-ray spectrometry and computing techniques are applied; thin-layer and paper radio chromatographic techniques for use with low energy beta-ray emitters; various aspects of liquid scintillation counting techniques in the measurement of alpha- and beta-ray emitters, including chemical and colour quenching; autoradiographic techniques; calibration of equipment; and standardization of radioisotopes. Finally, some applications of solid-state detectors are presented; this section may be regarded as a preview of important future developments. The meeting was attended by 203 participants
The Q sort theory and technique.
Nyatanga, L
1989-10-01
This paper is based on the author's experience of using the Q sort technique with BA Social Sciences (BASS) students, and the community psychiatric nursing (CPN, ENB No 811 course). The paper focuses on two main issues: 1. The theoretical assumptions underpinning the Q Sort technique. Carl Rogers' self theory and some of the values of humanistic psychology are summarised. 2. The actual technique procedure and meaning of results are highlighted. As the Q Sort technique is potentially useful in a variety of sittings some of which are listed in this paper, the emphasis has deliberately been placed in understanding the theoretical underpinning and the operationalisation (sensitive interpretation) of the theory to practice.
Vellinga, T P van Rees; Sterk, W; de Boer, A G E M; van der Beek, A J; Verhoeven, A C; van Dijk, F J H
2008-01-01
The Western Scheldt Tunneling Project in The Netherlands provided a unique opportunity to evaluate two deep-diving techniques with Doppler ultrasound surveillance. Divers used the bounce diving techniques for repair and maintenance of the TBM. The tunnel boring machine jammed at its deepest depth. As a result the work time was not sufficient. The saturation diving technique was developed and permitted longer work time at great depth. Thirty-one divers were involved in this project. Twenty-three divers were examined using Doppler ultrasound. Data analysis addressed 52 exposures to Trimix at 4.6-4.8 bar gauge using the bounce technique and 354 exposures to Trimix at 4.0-6.9 bar gauge on saturation excursions. No decompression incidents occurred with either technique during the described phase of the project. Doppler ultrasound revealed that the bubble loads assessed in both techniques were generally low. We find out, that despite longer working hours, shorter decompression times and larger physical workloads, the saturation-excursion technique was associated with significant lower bubble grades than in the bounce technique using Doppler Ultrasound. We conclude that the saturation-excursion technique with Trimix is a good option for deep and long exposures in caisson work. The Doppler technique proved valuable, and it should be incorporated in future compressed-air work.
Premature Ejaculation and Utilization of Cognitive Techniques
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Serkan AKKOYUNLU
2013-03-01
Full Text Available Introduction: Premature ejaculation is the most common male sexual dysfunction leading to distress in many couples. Master and Johnson emphasized the concept of early learned experiences and Kaplan emphasized lack of sensory awareness. For treatment sex therapists mainly utilize start-stop and squeeze techniques as homework. Couples enter sex therapy with some cognitive distortions and beliefs about sex and sexuality. These beliefs are also named sexual myths. For some couples using techniques to challenge cognitive distortions and maladaptive beliefs about sex and sexuality can be used. In this paper by presenting a case we discussed how cognitive techniques can be used along with behaviour techniques with couples. Case: Presenting clients are five years married couple who are thirty and twenty nine years old respectively. They attended to the outpatient clinic with the request of the female client. Their main complaint was premature ejaculation. They were diagnosed premature ejaculation using clinical interview. In treatment besides start and stop technique, cognitive techniques were utilized to address dysfunctional beliefs about sexuality. Discussion: Premature ejaculation is a male sexual dysfunction that causes distress and intimacy problems between couples. Stop start and squeeze techniques were accepted as the choice of treatment but their effectiveness is questioned recently. Also cognitive distortions and maladaptive beliefs may hamper therapy progress. Besides that, behavioral techniques utilizing cognitive techniques to lessen the degree of dysfunctional beliefs about sex and sexuality may help the couple to overcome premature ejaculation and enhance sexual satisfaction and intimacy.
Dual kidney transplant techniques: A systematic review.
Cocco, Annelise; Shahrestani, Sara; Cocco, Nicholas; Hameed, Ahmer; Yuen, Lawrence; Ryan, Brendan; Hawthorne, Wayne; Lam, Vincent; Pleass, Henry
2017-08-01
Dual kidney transplantation (DKT) was developed to improve outcomes from transplantation of extended criteria donors (ECD). This study examined which surgical techniques have been reported for DKT and whether any technique had superior patient and graft survival. Electronic databases were searched for published studies mapping to MESH terms: "kidney or renal" AND "transplan*" AND "dual or double." Single case reports, studies of patients less than 18 years old, studies which did not describe the surgical technique, and studies that did not report patient or graft survival were excluded. Fifteen reports of 434 DKT recipients were identified. Three techniques were described: bilateral placement; unilateral placement with separate anastomoses; and unilateral placement with patch anastomoses. Patient survival across all three techniques was over 95% at 1 year, and graft survival was also similar at over 90%. Rates of delayed graft function were between 20% and 30% across all techniques. The three techniques have equivalent delayed graft function as well as patient and graft survival rates. This is an encouraging result as it means that the surgeon can choose to use the technique which is most appropriate for their own skills and for the patient. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Modified grassline technique for orthodontic space closure.
Lohmiller, Rose Marie
2006-04-01
Use of traditional orthodontic measures in the periodontally compromised dentition can be problematic. This article describes a variation of the Grassline technique, in which the author uses adhesive bonding to harmoniously re-align malpositioned teeth. In this simple, easy-to-learn technique, a cotton thread is used to achieve orthodontic tooth movement. Initial contact with the saliva causes shrinkage of the thread, exercising a minor force that moves the teeth while allowing enough time for the tissues to regenerate. With this moderate, intermittent force, successful treatment is achieved with minimal risk. Moreover, this orthodontic technique has been associated with an increase in bone volume. This article presents this new technique and details methods for maintaining treatment success. The technique also is compared with an approach described in the literature. Two case reports are presented; the technique is employed in the first to close a diastema in a periodontally compromised dentition and in the second to correct the migration of maxillary and mandibular incisors. Photographs and radiographs show the esthetic improvement achieved in both cases using this technique.
Resolution enhancement techniques in microscopy
Cremer, Christoph; Masters, Barry R.
2013-05-01
We survey the history of resolution enhancement techniques in microscopy and their impact on current research in biomedicine. Often these techniques are labeled superresolution, or enhanced resolution microscopy, or light-optical nanoscopy. First, we introduce the development of diffraction theory in its relation to enhanced resolution; then we explore the foundations of resolution as expounded by the astronomers and the physicists and describe the conditions for which they apply. Then we elucidate Ernst Abbe's theory of optical formation in the microscope, and its experimental verification and dissemination to the world wide microscope communities. Second, we describe and compare the early techniques that can enhance the resolution of the microscope. Third, we present the historical development of various techniques that substantially enhance the optical resolution of the light microscope. These enhanced resolution techniques in their modern form constitute an active area of research with seminal applications in biology and medicine. Our historical survey of the field of resolution enhancement uncovers many examples of reinvention, rediscovery, and independent invention and development of similar proposals, concepts, techniques, and instruments. Attribution of credit is therefore confounded by the fact that for understandable reasons authors stress the achievements from their own research groups and sometimes obfuscate their contributions and the prior art of others. In some cases, attribution of credit is also made more complex by the fact that long term developments are difficult to allocate to a specific individual because of the many mutual connections often existing between sometimes fiercely competing, sometimes strongly collaborating groups. Since applications in biology and medicine have been a major driving force in the development of resolution enhancing approaches, we focus on the contribution of enhanced resolution to these fields.
Progress involving new techniques for liposome preparation
Directory of Open Access Journals (Sweden)
Zhenjun Huang
2014-08-01
Full Text Available The article presents a review of new techniques being used for the preparation of liposomes. A total of 28 publications were examined. In addition to the theories, characteristics and problems associated with traditional methods, the advantages and drawbacks of the latest techniques were reviewed. In the light of developments in many relevant areas, a variety of new techniques are being used for liposome preparation and each of these new technique has particular advantages over conventional preparation methods. However, there are still some problems associated with these new techniques that could hinder their applications and further improvements are needed. Generally speaking, due to the introduction of these latest techniques, liposome preparation is now an improved procedure. These applications promote not only advances in liposome research but also the methods for their production on an industrial scale.
Hlaallele Detection Using Molecular Techniques
Directory of Open Access Journals (Sweden)
Philip A. Dyer
1993-01-01
Full Text Available There are now many molecular biological techniques available to define HLA class I and class II alleles. Some of these are also applicable to other human polymorphic genes, in particular to those non-HLA genes encoded within the Mhc. The range of techniques available allows laboratories to choose those most suited to their purpose. The routine laboratory supporting solid organ transplants will need to type large numbers of potential recipients over a period of time, probably using PCR-SSOP while donors will be typed singly and rapidly using PCR-SSP with HLA allele compatibility determined by heteroduplex analysis. Laboratories supporting bone marrow transplantation, where time is less pressing, can choose from the whole range of techniques to determine accurately donor recipient Mhc compatibility. For disease studies, techniques defining precise HLA allele sequence polymorphisms are needed and high sample numbers have to be accommodated. When an association is established allele sequencing has to be used. In the near future, the precise role of HLA alleles in transplantation and disease susceptibility is likely to be established unambiguously.
The Algebraic Reconstruction Technique (ART)
International Nuclear Information System (INIS)
Raparia, D.; Alessi, J.; Kponou, A.
1997-01-01
Projections of charged particle beam current density (profiles) are frequently used as a measure of beam position and size. In conventional practice only two projections, usually horizontal and vertical, are measured. This puts a severe limit on the detail of information that can be achieved. A third projection provides a significant improvement. The Algebraic Reconstruction Technique (ART) uses three or more projections to reconstruct 3-dimensional density profiles. At the 200 MeV H-linac, we have used this technique to measure beam density, and it has proved very helpful, especially in helping determine if there is any coupling present in x-y phase space. We will present examples of measurements of current densities using this technique
New techniques in neutron scattering
International Nuclear Information System (INIS)
Hayter, J.B.
1993-01-01
New neutron sources being planned, such as the Advanced Neutron Source (ANS) or the European Spallation Source (ESS), will provide an order of magnitude flux increase over what is available today, but neutron scattering will still remain a signal-limited technique. At the same time, the development of new materials, such as polymer and ceramic composites or a variety of complex fluids, will increasingly require neutron-based research. This paper will discuss some of the new techniques which will allow us to make better use of the available neutrons, either through improved instrumentation or through sample manipulation. Discussion will center primarily on unpolarized neutron techniques since polarized neutrons will be the subject of the next paper. (author)
Directory of Open Access Journals (Sweden)
Elizabeth González Aznar
2017-02-01
Full Text Available Streptococcus pneumoniae (neumococo constituye una de las principales causas de enfermedades infecciosas bacterianas, particularmente en niños menores de 2 años de edad. Basadas en el polisacárido capsular (PsC, su principal factor de virulencia, existen dos tipos de vacunas aprobadas para uso en humanos: las vacunas polisacarídicas planas y las vacunas polisacarídicas conjugadas. Quimi-Vio, la vacuna antineumocócica cubana, pertenece al segundo grupo y está compuesta por los PsC de los siete serotipos de mayor incidencia y circulación en Cuba (1, 5, 6B, 14, 18C, 19F y 23F. El objetivo de este trabajo fue realizar el ensayo de identidad de la vacuna cubana Quimi-Vio empleando los AcM contra los PsC 1, 5, 6B, 14, y 19F de Sp obtenidos recientemente en el Instituto Finlay de Vacunas, teniendo en cuenta que los Ensayos de identidad de las vacunas son requisito indispensable para la liberación final de los lotes. La técnica empleada para la realización del ensayo de identidad de Quimi-Vio fue el Dot Blot, donde empleando membrana de Nitrocelulosa se realizó la captura de tres lotes de vacuna Quimi-Vio y como control positivo de la técnica los respectivos PsC de los serotipos 1, 5, 6B, 14, y 19F sin conjugar y una vacuna comercial Prevenar-13. Para la identidad de los PsC de neumococo se emplearon los respectivos AcM a una concentración de 10µg/mL y como segundo anticuerpo anti IgG de ratón conjugado a peroxidasa teniendo en cuenta que los AcMs empleados son murinos. Cada AcM fue capaz de identificar de forma altamente especifica al PsC homologo (mismo serotipo, tanto en su forma no conjugada y monovalente como en el contexto de las vacunas Quimi-Vio y Prevenar 13V, donde además de estar conjugado al TT se encuentra mezclado con otros PsC de forma multivalente. La técnica Dot Blot empleando los AcMs contra los PsC serotipos 1, 5,6B, 14 y 19F permite identificar de forma específica cada PsC en la formulación multivalente, por lo
Directory of Open Access Journals (Sweden)
Ho-Sook Song
2012-09-01
Full Text Available Purpose Different techniques for cystocele repair including the conventional anterior colporrhaphy and mesh technique are known. Our goal was to evaluate the anatomical success and safety of our method of transvaginal anterior vaginal wall repair by the purse-string technique reinforced with three simple additional sutures in the repair of cystocele over a 4-year follow-up period. Methods This was a retrospective review of 69 consecutive patients (grades 2 to 4 who underwent the above operations between 2001 and 2011, including their success rates as assessed by use of the Baden-Walker halfway classification system. Results Of the patients, 62 patients (98% were completely cured of cystocele and 1 patient showed grade 2 cystocele recurrence that required no further treatment. Two patients with grade 4 cystocele were completely cured. There was no vaginal erosion related to the cystocele repair. Conclusions Transvaginal anterior colporrhaphy by a purse-string technique reinforced with simple additive sutures appears to be a simple, safe, and easily performed approach in cystocele repair. There is no need for other material for reinforcement, even in high-grade cystocele, which is an advantage of our technique.
Magnetic characterization techniques for nanomaterials
2017-01-01
Sixth volume of a 40 volume series on nanoscience and nanotechnology, edited by the renowned scientist Challa S.S.R. Kumar. This handbook gives a comprehensive overview about Magnetic Characterization Techniques for Nanomaterials. Modern applications and state-of-the-art techniques are covered and make this volume an essential reading for research scientists in academia and industry.
Q-Technique and Graphics Research.
Kahle, Roger R.
Because Q-technique is as appropriate for use with visual and design items as for use with words, it is not stymied by the topics one is likely to encounter in graphics research. In particular Q-technique is suitable for studying the so-called "congeniality" of typography, for various copytesting usages, and for multivariate graphics research. The…
Stochastic Feedforward Control Technique
Halyo, Nesim
1990-01-01
Class of commanded trajectories modeled as stochastic process. Advanced Transport Operating Systems (ATOPS) research and development program conducted by NASA Langley Research Center aimed at developing capabilities for increases in capacities of airports, safe and accurate flight in adverse weather conditions including shear, winds, avoidance of wake vortexes, and reduced consumption of fuel. Advances in techniques for design of modern controls and increased capabilities of digital flight computers coupled with accurate guidance information from Microwave Landing System (MLS). Stochastic feedforward control technique developed within context of ATOPS program.
The Dermal Apron Technique for Immediate Implant Socket Management: A Novel Technique.
Levin, Barry P
2016-01-01
With immediate implant placement and provisionalization (IIP) in the esthetic zone, measures to counter hard and soft tissue loss are frequently necessary. To reduce the morbidity associated with bone and connective tissue procurement, various exogenous materials are utilized. The "Dermal Apron Technique" presented in this article demonstrates the use of a composite bone particulate (allograft/xenograft) plus a dermal allograft, adapted around screw-retained temporary crowns and secured within a subperiosteal pouch. The purpose is to augment the thickness of peri-implant mucosa for the purpose of preserving ridge dimensions and preventing mucosal recession. Controlled studies are required to further support its use. Clinical significance: Soft tissue health and harmony are critical for successful implant therapy in the esthetic regions of the dentition. Often, autogenous soft tissue grafts are used to augment peri-implant soft tissues. The Dermal Apron Technique is a method, that in specific situations, obviates the need for autogenous grafting. This reduces treatment time and morbidity associated with procurement of these grafts. The Dermal Apron Technique is used simultaneous with immediate placement and provisionalization and can improve long-term esthetic outcomes for patients. © 2016 Wiley Periodicals, Inc.
Development of hotcell non-destructive examination techniques
Energy Technology Data Exchange (ETDEWEB)
Kim, Young Uhn; Yu, S. C.; Kang, B. S.; Byun, K. S. [Chungbuk National University, Chungju (Korea)
2002-01-01
The purpose of this project is to establish non-destructive examination techniques which needs to determine the status of spent nuclear fuel and/or bundles. Through the project, we will establish an image reconstruction tomography which is a kind of non-destructive techniques in Hotcell. The tomography technique can be used to identify the 2-dimensional density distribution of fission products in the spent fuel rods and/or bundles. And form results of the measurement and analysis of magnetic properties of neutron irradiated material in the press vessel and reactor, we will develop some techniques to test its hardness and defects. In 2001, the first year, we have established mathematical background and necessary data and informations to develop the techniques. We will try to find some experimental results that are necessary in developing the Hotcell non-destructive examination techniques in the coming year. 14 refs., 65 figs., 5 tabs. (Author)
A new neutron noise technique for fast reactors
International Nuclear Information System (INIS)
Zhuo Fengguan; Jin Manyi; Yao Shigui; Su Zhuting
1987-12-01
This paper gives a new neutron noise technique for fast reactors, which is known as thermalization measurement technique of the neutron noise. The theoretical formulas of the technique were developed, and a digital delayed coincidence time analyzer consisted of TTL integrated circuits was constructed for the study of this technique. The technique has been tested and applied practically at Df-VI fast zero power reactor. It was shown that the provided technique in this work has a number of significant advantages in comparison with the conventional neutron noise method
Seminar on Detectors and measurements techniques
International Nuclear Information System (INIS)
Holm, E.
2002-01-01
A Nordic Seminar on detectors and radionuclide measurement techniques was held in Lund, Sweden, May 3-4, 2001. The objective was to highlight recent progress and problems for techniques to study environmental radioactivity. It covered the aspect of detector sample geometry's and methods for evaluation of gamma gamma pulse height distributions. Within the field of alpha-spectrometric techniques gridded ionisation chambers, semiconductor detectors and a general description for analysis of alpha-particle-spectra were presented. Recent development in mass spectrometric techniques, AMS (Accelerator Mass Spectrometry) and ICPMS (Inductively Coupled Plasma mass Spectrometry) for long-lived radionuclides was described. Principles for analysis of beta particle emitters, especially by liquid scintillation were presented. The seminar also covered radiochemistry such advantages and disadvantages between ion exchange, solvent extraction and extraction chromatography. The use of controlled laboratory conditions for discerning the dynamics of accumulation in organisms was demonstrated. Other techniques such as neutron activation were also shown to be useful analytical tool for certain long-lived radionuclides. The results of the intercalibration exercises within the Nordic countries showed the importance of such analytical quality control. (au)
Dalbayrak, Sedat; Yaman, Onur; Yılmaz, Mesut
2013-07-01
Treatment of Hangman's fractures is still controversial. Hangman's fractures Type II and IIA are usually treated with surgical procedures. This study aims at describing the Neurospinal Academy (NSA) technique as an attempt to achieve an approximation of the fracture line to the axis body, which may be used for Type II and IIA patients with severe displacement and angulation. NSA technique both pars or pedicle screws are placed bicortically to ensure that anterior surface of C2 vertebral body will be crossed 1-2 mm. A rod is prepared in suitable length and curve to connect the two screws. For placing the rod, sufficient amount of bone is resected from the C2 spinous process. C2 vertebral body is pulled back by means of the screws that crossed the anterior surface of C2 vertebral body. Hangman II and IIA patient are treated with NSA technique. Angulated and tilted C2 vertebral body was pulled back and approximated to posterior elements. In Hangman's fractures Type II and IIA with severe vertebral body and pedicle displacement, NSA technique is an effective and reliable treatment alternative for the approximation of posterior elements to the C2 vertebral body, which is tilted, angulated, and dislocated.
Studies on antigenic cross-reactivity of Trichuris ovis with host mucosal antigens in goat
Gautam Patra; Seikh Sahanawaz Alam; Sonjoy Kumar Borthakur; Hridayesh Prasad
2015-01-01
Objective: To ascertain whether immunodominant antigens of Trichuris ovis might share and cross react with host molecule. Methods: Two crude protein preparations from anterior and posterior parts of Trichuris ovis were characterized along with host mucosal antigen by double immunodiffusion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotting technique. Conventional scanning electron microscopy was performed as per standard procedure. Results: Sharp...
Magnetic Nondestructive Testing Techniques of Constructional Steel
Directory of Open Access Journals (Sweden)
Xiong Er-gang
2016-01-01
Full Text Available Steel is a kind of ferromagnetic material, which is extensively applied in such fields as buildings, bridges, railways, machines and lifeline engineering etc. Those engineering structures built of constructional steel will unavoidably experience some damages during their service lifetime, thus which will influence the distribution regularity of internal forces in structures, result in over-stresses, cause the local failure of structures, and even lead to collapse of the whole structure. Therefore, it is a pressing topic to study how to directly evaluate the real-time stressed states of structural members, damages and steel characteristics in present structural health monitoring and diagnosing fields. And the achievements of this research will be of theoretical significance and of application value of engineering. This paper summarizes varieties of new magnetic nondestructive testing techniques used in constructional steel, respectively investigates the testing principles, characteristics and application for the magnetic Barkhausen noise technique, magnetic acoustic emission technique, magnetic flux leakage technique, magnetic memory technique and magnetic absorption technique, and points out the problems present in the application of these new techniques to actual testing and the further research objective.
Computer Assisted Audit Techniques
Directory of Open Access Journals (Sweden)
Eugenia Iancu
2007-01-01
Full Text Available From the modern point of view, audit takes intoaccount especially the information systems representingmainly the examination performed by a professional asregards the manner for developing an activity by means ofcomparing it to the quality criteria specific to this activity.Having as reference point this very general definition ofauditing, it must be emphasized that the best known segmentof auditing is the financial audit that had a parallel evolutionto the accountancy one.The present day phase of developing the financial audithas as main trait the internationalization of the accountantprofessional. World wide there are multinational companiesthat offer services in the financial auditing, taxing andconsultancy domain. The auditors, natural persons and auditcompanies, take part at the works of the national andinternational authorities for setting out norms in theaccountancy and auditing domain.The computer assisted audit techniques can be classified inseveral manners according to the approaches used by theauditor. The most well-known techniques are comprised inthe following categories: testing data techniques, integratedtest, parallel simulation, revising the program logics,programs developed upon request, generalized auditsoftware, utility programs and expert systems.
Fundamentals - state of the art of radiation techniques
International Nuclear Information System (INIS)
Wogman, N.A.
1982-01-01
In minerals exploration and extraction, nuclear techniques have several advantages. The techniques are elementally specific and their exploration range varies from a few millimeters in average rock formations to more than a meter. Because of the heterogeneous disposition of minerals and difficult environments in which measurements are required (in boreholes, on conveyor belts, in bunkers), interrogating techniques are required which exhibit both elemental specificity and range. It is for these fundamental reasons that nuclear techniques are the only possible techniques which satisfy all requirements. A variety of techniques have been developed and used. These are based on energy dispersive x-ray fluorescence (EDXRF), measurement of natural gamma-ray radiation, gamma-ray attenuation and scattering, and on neutron interactions. This paper discusses the fundamentals of these four techniques and their applications. A table is also provided listing some existing selected applications of nuclear techniques in mineral exploration, mining and processing
Overview of nonchemical decontamination techniques
International Nuclear Information System (INIS)
Allen, R.P.
1984-09-01
The decontamination techniques summarized in this paper represent a variety of surface cleaning methods developed or adapted for component and facility-type decontamination applications ranging from small hand tools to reactor cavities and other large surface areas. The major conclusion is that decontamination is a complex, demanding technical discipline. It requires knowledgeable, experienced and well-trained personnel to select proper techniques and combinations of techniques for the varied plant applications and to realize their full performance potential. Unfortunately, decontamination in many plants has the lowest priority of almost any activity. Operators are unskilled and turnover is so frequent that expensive decontamination capabilities remain unused while decontamination operations revert to the most rudimentary type of hand scrubbing and water spray cleaning
International Nuclear Information System (INIS)
Anon.
1976-01-01
The report reviews radon measurement surveys in soils and in water. Special applications, and advantages and limitations of the radon measurement techniques are considered. The working group also gives some directions for further research in this field
International Nuclear Information System (INIS)
Bernstein, J.M.; Stokes, C.E.; Fernie, B.
1987-01-01
Immunoblotting is a powerful technique for the detection of small amounts of immunologically interesting proteins in unpurified preparations. Iodinated protein A (PA) has been widely used as a second antibody for detection of proteins; however, it does not bind equally well to immunoglobulins from different species nor does it bind to all subclasses of immunoglobulin G (IgG). We compared the sensitivity of [ 125 I]PA with those of both horseradish peroxidase-conjugated second antibodies (HRP) and glucose oxidase-anti-glucose oxidase (GAG) soluble complexes for visualizing bovine serum albumin, human IgG, or human C3 which was either dot blotted or electroblotted to nitrocellulose. [ 125 I]PA was uniformly 10- to 100-fold less sensitive than either HRP or GAG. GAG was more sensitive than HRP except for C3 (electroblotting) and bovine serum albumin and IgG (dot blotting), in which they were equivalent. In general, dot blotting was 10- to 1000-fold more sensitive than electroblotting. Although relative sensitivities varied depending on the proteins analyzed and the antisera used, GAG appeared to be superior to [ 125 I]PA and HRP for detection of immunoblotted proteins
An anonymization technique using intersected decision trees
Directory of Open Access Journals (Sweden)
Sam Fletcher
2015-07-01
Full Text Available Data mining plays an important role in analyzing the massive amount of data collected in today’s world. However, due to the public’s rising awareness of privacy and lack of trust in organizations, suitable Privacy Preserving Data Mining (PPDM techniques have become vital. A PPDM technique provides individual privacy while allowing useful data mining. We present a novel noise addition technique called Forest Framework, two novel data quality evaluation techniques called EDUDS and EDUSC, and a security evaluation technique called SERS. Forest Framework builds a decision forest from a dataset and preserves all the patterns (logic rules of the forest while adding noise to the dataset. We compare Forest Framework to its predecessor, Framework, and another established technique, GADP. Our comparison is done using our three evaluation criteria, as well as Prediction Accuracy. Our experimental results demonstrate the success of our proposed extensions to Framework and the usefulness of our evaluation criteria.
[Intestinal lengthening techniques: an experimental model in dogs].
Garibay González, Francisco; Díaz Martínez, Daniel Alberto; Valencia Flores, Alejandro; González Hernández, Miguel Angel
2005-01-01
To compare two intestinal lengthening procedures in an experimental dog model. Intestinal lengthening is one of the methods for gastrointestinal reconstruction used for treatment of short bowel syndrome. The modification to the Bianchi's technique is an alternative. The modified technique decreases the number of anastomoses to a single one, thus reducing the risk of leaks and strictures. To our knowledge there is not any clinical or experimental report that studied both techniques, so we realized the present report. Twelve creole dogs were operated with the Bianchi technique for intestinal lengthening (group A) and other 12 creole dogs from the same race and weight were operated by the modified technique (Group B). Both groups were compared in relation to operating time, difficulties in technique, cost, intestinal lengthening and anastomoses diameter. There were no statistical difference in the anastomoses diameter (A = 9.0 mm vs. B = 8.5 mm, p = 0.3846). Operating time (142 min vs. 63 min) cost and technique difficulties were lower in group B (p anastomoses (of Group B) and intestinal segments had good blood supply and were patent along their full length. Bianchi technique and the modified technique offer two good reliable alternatives for the treatment of short bowel syndrome. The modified technique improved operating time, cost and technical issues.
Magnetic field measurements and mapping techniques
CERN. Geneva
2003-01-01
These lectures will present an overview of the most common techniques used for the measurement of magnetic field in accelerator magnets. The formalism for a harmonic description of the magnetic field will be presented, including a discussion of harmonics allowed under various types of symmetries in the magnet. The harmonic coil technique for measurement of field harmonics will be covered in depth. Using examples from recent projects, magnetic measurements will be shown to be a powerful tool for monitoring magnet production. Measurements of magnetic axis using extensions of the harmonic coil technique, as well as other techniques, such as the colloidal cell and stretched wire, will be covered. Topics of interest in superconducting magnets, such as time decay and snapback, requiring relatively fast measurements of the harmonics, will also be described.
Soft computing techniques in engineering applications
Zhong, Baojiang
2014-01-01
The Soft Computing techniques, which are based on the information processing of biological systems are now massively used in the area of pattern recognition, making prediction & planning, as well as acting on the environment. Ideally speaking, soft computing is not a subject of homogeneous concepts and techniques; rather, it is an amalgamation of distinct methods that confirms to its guiding principle. At present, the main aim of soft computing is to exploit the tolerance for imprecision and uncertainty to achieve tractability, robustness and low solutions cost. The principal constituents of soft computing techniques are probabilistic reasoning, fuzzy logic, neuro-computing, genetic algorithms, belief networks, chaotic systems, as well as learning theory. This book covers contributions from various authors to demonstrate the use of soft computing techniques in various applications of engineering.
Integral transform techniques for Green's function
Watanabe, Kazumi
2015-01-01
This book describes mathematical techniques for integral transforms in a detailed but concise manner. The techniques are subsequently applied to the standard partial differential equations, such as the Laplace equation, the wave equation and elasticity equations. Green’s functions for beams, plates and acoustic media are also shown, along with their mathematical derivations. The Cagniard-de Hoop method for double inversion is described in detail, and 2D and 3D elastodynamic problems are treated in full. This new edition explains in detail how to introduce the branch cut for the multi-valued square root function. Further, an exact closed form Green’s function for torsional waves is presented, as well as an application technique of the complex integral, which includes the square root function and an application technique of the complex integral.
New techniques for wound debridement.
Madhok, Brijesh M; Vowden, Kathryn; Vowden, Peter
2013-06-01
Debridement is a crucial component of wound management. Traditionally, several types of wound debridement techniques have been used in clinical practice such as autolytic, enzymatic, biodebridement, mechanical, conservative sharp and surgical. Various factors determine the method of choice for debridement for a particular wound such as suitability to the patient, the type of wound, its anatomical location and the extent of debridement required. Recently developed products are beginning to challenge traditional techniques that are currently used in wound bed preparation. The purpose of this review was to critically evaluate the current evidence behind the use of these newer techniques in clinical practice. There is some evidence to suggest that low frequency ultrasound therapy may improve healing rates in patients with venous ulcers and diabetic foot ulcers. Hydrosurgery debridement is quick and precise, but the current evidence is limited and further studies are underway. Debridement using a monofilament polyester fibre pad and plasma-mediated bipolar radiofrequency ablation are both very new techniques. The initial evidence is limited, and further studies are warranted to confirm their role in management of chronic wounds. © 2013 The Authors. International Wound Journal © 2013 John Wiley & Sons Ltd and Medicalhelplines.com Inc.
International Nuclear Information System (INIS)
Ercan, T.; Alco, G.; Zengin, F.; Atilla, S.; Dincer, M.; Igdem, S.; Okkan, S.
2010-01-01
The aim of this study was to be able to implement the field-in-field intensity-modulated radiotherapy (FiF) technique in our daily practice for breast radiotherapy. To do this, we performed a dosimetric comparison. Treatment plans were produced for 20 consecutive patients. FiF plans and conformal radiotherapy (CRT) plans were compared for doses in the planning target volume (PTV), the dose homogeneity index (DHI), doses in irradiated soft tissue outside the target volume (SST), ipsilateral lung and heart doses for left breast irradiation, and the monitor unit counts (MU) required for treatment. Averaged values were compared using Student's t-test. With FiF, the DHI is improved 7.0% and 5.7%, respectively (P<0.0001) over the bilateral and lateral wedge CRT techniques. When the targeted volumes received 105% and 110% of the prescribed dose in the PTV were compared, significant decreases are found with the FiF technique. With the 105% dose, the SST, heart, and ipsilateral lung doses and the MU counts were also significantly lower with the FiF technique. The FiF technique, compared to CRT, for breast radiotherapy enables significantly better dose distribution in the PTV. Significant differences are also found for soft tissue volume, the ipsilateral lung dose, and the heart dose. Considering the decreased MUs needed for treatment, the FiF technique is preferred over tangential CRT. (author)
Advanced Techniques in Biophysics
Arrondo, José Luis R
2006-01-01
Technical advancements are basic elements in our life. In biophysical studies, new applications and improvements in well-established techniques are being implemented every day. This book deals with advancements produced not only from a technical point of view, but also from new approaches that are being taken in the study of biophysical samples, such as nanotechniques or single-cell measurements. This book constitutes a privileged observatory for reviewing novel applications of biophysical techniques that can help the reader enter an area where the technology is progressing quickly and where a comprehensive explanation is not always to be found.
Nonenhanced MR angiography techniques
International Nuclear Information System (INIS)
Lanzman, R.S.; Kroepil, P.; Blondin, D.; Schmitt, P.
2011-01-01
Especially in regard to the potential risks for the development of nephrogenic systemic fibrosis (NSF) following the administration of Gadolinium-based contrast material, nonenhanced MR angiography (MRA) methods are becoming ever more important. Besides well-established time-of-flight (TOF) and phase-contrast (PC) MRA, alternative imaging techniques based on balanced steady-state free precession (bSSFP) and turbo-spin-echo (TSE) sequences are increasingly used in combination with or without arterial spin labeling (ASL) strategies. This article provides an overview of the principles and clinical values of different nonenhanced MRA techniques. In addition, recent nonenhanced MRA developments are presented. (orig.)
Materials Science and X-ray Techniques
International Nuclear Information System (INIS)
Brock, J.; Sutton, M.
2008-01-01
Many novel synchrotron-based X-ray techniques directly address the core questions of modern materials science but are not yet at the stage of being easy to use because of the lack of dedicated beamlines optimized for specific measurements. In this article, we highlight a few of these X-ray techniques and discuss why, with ongoing upgrades of existing synchrotrons and with new linear-accelerator-based sources under development, now is the time to ensure that these techniques are readily available to the larger materials research community.
Alternative Assessment Techniques.
Lowenthal, Barbara
1988-01-01
Maintaining the precision necessary for administering norm referenced tests can be a problem for the special education teacher who is trained to assist the student. Criterion-referenced tests, observations, and interviews are presented as effective alternative assessment techniques. (JDD)
Application of functional analysis techniques to supervisory systems
International Nuclear Information System (INIS)
Lambert, Manuel; Riera, Bernard; Martel, Gregory
1999-01-01
The aim of this paper is to apply firstly two interesting functional analysis techniques for the design of supervisory systems for complex processes, and secondly to discuss the strength and the weaknesses of each of them. Two functional analysis techniques have been applied, SADT (Structured Analysis and Design Technique) and FAST (Functional Analysis System Technique) on a process, an example of a Water Supply Process Control (WSPC) system. These techniques allow a functional description of industrial processes. The paper briefly discusses the functions of a supervisory system and some advantages of the application of functional analysis for the design of a 'human' centered supervisory system. Then the basic principles of the two techniques applied on the WSPC system are presented. Finally, the different results obtained from the two techniques are discussed
Malycha, Charlotte P.; Maier, Günter W.
2017-01-01
Although creativity techniques are highly recommended in working environments, their effects have been scarcely investigated. Two cognitive processes are often considered to foster creative potential and are, therefore, taken as a basis for creativity techniques: knowledge activation and conceptual combination. In this study, both processes were…
Soil analysis. Modern instrumental technique
International Nuclear Information System (INIS)
Smith, K.A.
1993-01-01
This book covers traditional methods of analysis and specialist monographs on individual instrumental techniques, which are usually not written with soil or plant analysis specifically in mind. The principles of the techniques are combined with discussions of sample preparation and matrix problems, and critical reviews of applications in soil science and related disciplines. Individual chapters are processed separately for inclusion in the appropriate data bases
Cicinelli, Ettore; Mitsopoulos, Vasileios; Fascilla, Fabiana D; Sioutis, Dimos; Bettocchi, Stefano
2016-06-01
Uterine fibroids, also known as leiomyomas, represent the most common benign tumors of the female genital tract. Submucosal leiomyomas are classified into three grades: G0, GI, GII according to the degree of their intramural proportion. A recently developed technique enables the preparation of G1 and G2 leiomyomas for their subsequent successful resection in a second step. The OPPIuM (office preparation of partially intramural leiomyomas) technique aims to downgrade type I and II leiomyomas, in order to facilitate a subsequent easier and safer resectoscopy. Hysteroscopic resection of large GI or GII submucosal fibroids is a complex procedure. OPPIuM technique has been invented and seems to achieve the downgrading of these types of leiomyomas in approximately 93% of cases, without any significant surgical complications or the need of hormonal agents' administration. In this way, the safer and quicker subsequent complete myomectomy is facilitated.
Vasectomy occlusion techniques for male sterilization.
Cook, L A; Vliet, H; Pun, A; Gallo, M F
2004-01-01
Vasectomy is an increasingly popular and effective family planning method. A variety of vasectomy techniques are used worldwide including various vas occlusion techniques (excision and ligation, thermal or electrocautery, and mechanical and chemical occlusion methods), vas irrigation and fascial interposition. Vasectomy guidelines largely rely on information from observational studies. Ideally, the choice of vasectomy techniques should be based on the best available evidence from randomized controlled trials. The objective of this review was to compare the effectiveness, safety, acceptability and costs of vasectomy techniques for male sterilization. We searched the computerized databases the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, Popline and LILACS. In addition, we searched the reference lists of relevant articles and book chapters. We included randomized controlled trials and controlled clinical trials comparing vasectomy techniques. We assessed all titles and abstracts located in the literature searches and two reviewers independently extracted articles identified for inclusion. Data were presented in the text of the review. Outcome measures include contraceptive efficacy, safety, discontinuation, and acceptability. Two trials compared vas occlusion with clips versus a conventional vasectomy technique; both were of poor quality. Neither trial found a difference between the two groups with regard to the primary outcome of failure to reach azoospermia. Four trials examined vas irrigation: three compared water irrigation with no irrigation and one compared water irrigation with euflavine. All of the trials were of poor quality. None of the trials found a significant difference between the groups with respect to the primary outcome of time to azoospermia. However, one trial found that the median number of ejaculations to azoospermia was significantly lower in the euflavine group compared to the water irrigation group. The one trial
Seminar on Detectors and measurements techniques
Energy Technology Data Exchange (ETDEWEB)
Holm, E. (ed.) [Risoe National Lab., Roskilde (Denmark)
2002-07-01
A Nordic Seminar on detectors and radionuclide measurement techniques was held in Lund, Sweden, May 3-4, 2001. The objective was to highlight recent progress and problems for techniques to study environmental radioactivity. It covered the aspect of detector sample geometry's and methods for evaluation of gamma gamma pulse height distributions. Within the field of alpha-spectrometric techniques gridded ionisation chambers, semiconductor detectors and a general description for analysis of alpha-particle-spectra were presented. Recent development in mass spectrometric techniques, AMS (Accelerator Mass Spectrometry) and ICPMS (Inductively Coupled Plasma mass Spectrometry) for long-lived radionuclides was described. Principles for analysis of beta particle emitters, especially by liquid scintillation were presented. The seminar also covered radiochemistry such advantages and disadvantages between ion exchange, solvent extraction and extraction chromatography. The use of controlled laboratory conditions for discerning the dynamics of accumulation in organisms was demonstrated. Other techniques such as neutron activation were also shown to be useful analytical tool for certain long-lived radionuclides. The results of the intercalibration exercises within the Nordic countries showed the importance of such analytical quality control. (au)
New computing techniques in physics research
International Nuclear Information System (INIS)
Becks, Karl-Heinz; Perret-Gallix, Denis
1994-01-01
New techniques were highlighted by the ''Third International Workshop on Software Engineering, Artificial Intelligence and Expert Systems for High Energy and Nuclear Physics'' in Oberammergau, Bavaria, Germany, from October 4 to 8. It was the third workshop in the series; the first was held in Lyon in 1990 and the second at France-Telecom site near La Londe les Maures in 1992. This series of workshops covers a broad spectrum of problems. New, highly sophisticated experiments demand new techniques in computing, in hardware as well as in software. Software Engineering Techniques could in principle satisfy the needs for forthcoming accelerator experiments. The growing complexity of detector systems demands new techniques in experimental error diagnosis and repair suggestions; Expert Systems seem to offer a way of assisting the experimental crew during data-taking
Artificial intelligence techniques in power systems
Energy Technology Data Exchange (ETDEWEB)
Laughton, M.A.
1997-12-31
Since the early to mid 1980s much of the effort in power systems analysis has turned away from the methodology of formal mathematical modelling which came from the fields of operations research, control theory and numerical analysis to the less rigorous techniques of artificial intelligence (AI). Today the main AI techniques found in power systems applications are those utilising the logic and knowledge representations of expert systems, fuzzy systems, artificial neural networks (ANN) and, more recently, evolutionary computing. These techniques will be outlined in this chapter and the power system applications indicated. (Author)
New CT-aided stereotactic neurosurgery technique
International Nuclear Information System (INIS)
Shao, H.M.; Truong, T.K.; Reed, I.S.; Slater, R.A.
1985-01-01
In this communication, a new technique for CT-aided stereotactic neurosurgery is presented. The combination of specially designed hardware and software provides a fast, simple, and versatile tool for the accurate insertion of a probe into the human brain. This system is portable and can be implemented on any CT computer system. The complete procedure to perform the CT-aided stereotactic neurosurgery technique is presented. Experimental results are given which demonstrate the power of the method. Finally, the key algorithms for realizing this technique are described in the Appendix
GPU PRO 3 Advanced rendering techniques
Engel, Wolfgang
2012-01-01
GPU Pro3, the third volume in the GPU Pro book series, offers practical tips and techniques for creating real-time graphics that are useful to beginners and seasoned game and graphics programmers alike. Section editors Wolfgang Engel, Christopher Oat, Carsten Dachsbacher, Wessam Bahnassi, and Sebastien St-Laurent have once again brought together a high-quality collection of cutting-edge techniques for advanced GPU programming. With contributions by more than 50 experts, GPU Pro3: Advanced Rendering Techniques covers battle-tested tips and tricks for creating interesting geometry, realistic sha
Design techniques for mutlivariable flight control systems
1981-01-01
Techniques which address the multi-input closely coupled nature of advanced flight control applications and digital implementation issues are described and illustrated through flight control examples. The techniques described seek to exploit the advantages of traditional techniques in treating conventional feedback control design specifications and the simplicity of modern approaches for multivariable control system design.
A simplified technique for shakedown load determination
International Nuclear Information System (INIS)
Abdalla, H.F.; Younan, M.Y.A.; Megahed, M.M.
2005-01-01
In this paper a simple technique is presented to determine the limit shakedown load of a structure or a component using the finite element method. Through the proposed technique, the limit shakedown load is determined without performing time consuming cyclic loading simulations or iterative elastic techniques. Instead, it is determined by performing only two analyses namely, an elastic analysis and an elastic-plastic analysis. By extracting the results of the two analyses, the limit shakedown load of the structure is determined through the calculation of the residual stresses. The technique is applied and verified using two bench mark shakedown problems namely: the two-bar structure subjected to constant axial force and cyclic thermal loading, and the Bree cylinder subjected to constant internal pressure and cyclic high heat fluxes across its wall. The results of the proposed technique showed very good correlation with the, analytically determined, Bree diagrams of both structures. Moreover, the outcomes of the proposed technique showed very good results in comparison to full cyclic loading elasto-plastic finite element simulations of both structures. (authors)
Techniques for freeing deposited canisters. Final report
International Nuclear Information System (INIS)
Kalbantner, P.; Sjoeblom, R.
2000-06-01
Four different techniques for removal of the bentonite buffer around a deposited canister have been identified, studied and evaluated: mechanical, hydrodynamical, thermal, and electrical techniques. Different techniques to determine the position of the canister in the buffer have also been studied: mechanical, electromagnetic, thermal and acoustic techniques. The mechanical techniques studied are full-face boring, milling and core-drilling. It is expected that the bentonite can be machined relatively easily. It is assessed that cooling by means of flushing water over the outer surfaces of the tools is not feasible in view of the tendency of bentonite to form a gel. The mechanical techniques are characterized by the potential of damaging the canister, a high degree of complexity, and high requirements of energy/power input. The generated byproduct is solid and cannot be removed by means of flushing. Removal is assessed to be simplest in conjunction with full-face boring and most difficult when coredrilling is applied. The hydrodynamical techniques comprise high-pressure hydrodynamic techniques, where pressures above and below 100 bar, and low pressure hydrodynamical techniques (< 10 bar) are separated. At pressures above 100 bar, a water jet with a diameter of approximately a millimetre cuts through the material. If desired, sand can be added to the jet. At pressures below 100 bar the jet has a diameter of one or a few centimetres. The liquid contains a few percent of salt, which is essential for the efficiency of the process. The flushing is important not only because it removes the modified bentonite but also because it frees previously unaffected bentonite and thereby makes it accessible to chemical modification. All of the hydrodynamical techniques are applicable for freeing the end surface as well as the mantle surface. The degree of complexity and the requirement on energy/power decrease with a decrease in pressure. A significant potential for damaging the
Microdialysis technique and interventional radiology
International Nuclear Information System (INIS)
An Xiao; Xiao Xiangsheng
2007-01-01
Basic research in interventional radiology, including transcatheter artery perfusion especially, is progressing slowly due to lack of proper method. Microdialysis technique, a kind of accurate sampling technique in vivo, may help to solve the problem. Just as its name implies, microdialysis means tiny dialysis with advantages of authenticity, exactness and less error. Furthermore it has been applied widely and should be received with great attention and popularity. (authors)
Dumas, A.
1981-01-01
Three major areas that are considered in the development of an overall maintenance scheme of computer equipment are described. The areas of concern related to fault isolation techniques are: the programmer (or user), company and its policies, and the manufacturer of the equipment.
Multiband RF circuits and techniques for wireless transmitters
Chen, Wenhua; Ghannouchi, Fadhel M
2016-01-01
This book introduces systematic design methods for passive and active RF circuits and techniques, including state-of-the-art digital enhancement techniques. As the very first book dedicated to multiband RF circuits and techniques, this work provides an overview of the evolution of transmitter architecture and discusses current digital predistortion techniques. Readers will find a collection of novel research ideas and new architectures in concurrent multiband power dividers, power amplifiers and related digital enhancement techniques. This book will be of great interest to academic researchers, R&D engineers, wireless transmitter and protocol designers, as well as graduate students who wish to learn the core architectures, principles and methods of multiband RF circuits and techniques. .