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Sample records for labeled water method

  1. Labeling lake water with tritium

    USGS Publications Warehouse

    Frederick, B.J.

    1963-01-01

    A method of packaging tritiated water in a manner that facilitates safe handling in environmental labeling operations, and procedures followed in labeling a large body of water with a small volume of tritiated water are described. ?? 1963.

  2. Visualizing Water Molecules in Transmembrane Proteins Using Radiolytic Labeling Methods

    SciTech Connect

    Orban, T.; Gupta, S; Palczewski, K; Chance, M

    2010-01-01

    Essential to cells and their organelles, water is both shuttled to where it is needed and trapped within cellular compartments and structures. Moreover, ordered waters within protein structures often colocalize with strategically placed polar or charged groups critical for protein function, yet it is unclear if these ordered water molecules provide structural stabilization, mediate conformational changes in signaling, neutralize charged residues, or carry out a combination of all these functions. Structures of many integral membrane proteins, including G protein-coupled receptors (GPCRs), reveal the presence of ordered water molecules that may act like prosthetic groups in a manner quite unlike bulk water. Identification of 'ordered' waters within a crystalline protein structure requires sufficient occupancy of water to enable its detection in the protein's X-ray diffraction pattern, and thus, the observed waters likely represent a subset of tightly bound functional waters. In this review, we highlight recent studies that suggest the structures of ordered waters within GPCRs are as conserved (and thus as important) as conserved side chains. In addition, methods of radiolysis, coupled to structural mass spectrometry (protein footprinting), reveal dynamic changes in water structure that mediate transmembrane signaling. The idea of water as a prosthetic group mediating chemical reaction dynamics is not new in fields such as catalysis. However, the concept of water as a mediator of conformational dynamics in signaling is just emerging, because of advances in both crystallographic structure determination and new methods of protein footprinting. Although oil and water do not mix, understanding the roles of water is essential to understanding the function of membrane proteins.

  3. The Doubly Labeled Water Method for Measuring Human Energy Expenditure: Adaptations for Spaceflight

    NASA Technical Reports Server (NTRS)

    Schulz, Leslie O.

    1991-01-01

    It is essential to determine human energy requirements in space, and the doubly labeled water method has been identified as the most appropriate means of indirect calorimetry to meet this need. The method employs naturally occurring, stable isotopes of hydrogen (H-2, deuterium) and oxygen (O-18) which, after dosing, mix with body water. The deuterium is lost from the body as water while the O-18 is eliminated as both water and CO2. The difference between the two isotope elimination rates is therefore a measure of CO2 production and hence energy expenditure. Spaceflight will present a unique challenge to the application of the doubly labeled water method. Specifically, interpretation of doubly labeled water results assumes that the natural abundance or 'background' levels of the isotopes remain constant during the measurement interval. To address this issue, an equilibration model will be developed in an ongoing ground-based study. As energy requirements of women matched to counterparts in the Astronauts Corps are being determined by doubly labeled water, the baseline isotope concentration will be changed by consumption of 'simulated Shuttle water' which is artificially enriched. One group of subjects will be equilibrated on simulated Shuttle water prior to energy determinations by doubly labeled water while the others will consume simulated Shuttle water after dosing. This process will allow us to derive a prediction equation to mathematically model the effect of changing background isotope concentrations.

  4. Adaptation of the doubly labeled water method for subjects consuming isotopically enriched water.

    PubMed

    Gretebeck, R J; Schoeller, D A; Socki, R A; Davis-Street, J; Gibson, E K; Schulz, L O; Lane, H W

    1997-02-01

    The use of doubly labeled water (DLW) to measure energy expenditure is subject to error if the background abundance of the oxygen and hydrogen isotope tracers changes during the test period. This study evaluated the accuracy and precision of different methods by which such background isotope changes can be corrected, including a modified method that allows prediction of the baseline that would be achieved if subjects were to consume water from a given source indefinitely. Subjects in this study were eight women (4 test subjects and 4 control subjects) who consumed for 28 days water enriched to resemble drinking water aboard the United States space shuttle. Test subjects and control subjects were given a DLW dose on days 1 and 15, respectively. The change to an enriched water source produced a bias in expenditure calculations that exceeded 2.9 MJ/day (35%), relative to calculations from intake-balance. The proposed correction based on the predicted final abundance of 18O and deuterium after equilibration to the new water source eliminated this bias, as did the traditional use of a control group. This new modified correction method is advantageous under field conditions when subject numbers are limited.

  5. The doubly labeled water method produces highly reproducible longitudinal results in nutrition studies

    USDA-ARS?s Scientific Manuscript database

    The doubly labeled water (DLW) method is considered the reference method for the measurement of energy expenditure under free-living conditions. However, the reproducibility of the DLW method in longitudinal studies is not well documented. This study was designed to evaluate the longitudinal reprodu...

  6. Sensitivity of methods for calculating energy expenditure by use of doubly labeled water

    SciTech Connect

    Seale, J.; Miles, C.; Bodwell, C.E.

    1989-02-01

    Attempts to estimate human energy expenditure by use of doubly labeled water have produced three methods currently used for calculating carbon dioxide production from isotope disappearance data: (1) the two-point method, (2) the regression method, and (3) the integration method. An ideal data set was used to determine the error produced in the calculated energy expenditure for each method when specific variables were perturbed. The analysis indicates that some of the calculation methods are more susceptible to perturbations in certain variables than others. Results from an experiment on one adult human subject are used to illustrate the potential for error in actual data. Samples of second void urine, 24-h urine, and breath collected every other day for 21 days are used to calculate the average daily energy expenditure by three calculation methods. The difference between calculated energy expenditure and metabolizable energy on a weight-maintenance diet is used to estimate the error associated with the doubly labeled water method.

  7. Doubly labeled water method: in vivo oxygen and hydrogen isotope fractionation

    SciTech Connect

    Schoeller, D.A.; Leitch, C.A.; Brown, C.

    1986-12-01

    The accuracy and precision of the doubly labeled water method for measuring energy expenditure are influenced by isotope fractionation during evaporative water loss and CO/sub 2/ excretion. To characterize in vivo isotope fractionation, we collected and isotopically analyzed physiological fluids and gases. Breath and transcutaneous water vapor were isotopically fractionated. The degree of fractionation indicated that the former was fractionated under equilibrium control at 37/sup 0/C, and the latter was kinetically fractionated. Sweat and urine were unfractionated. By use of isotopic balance models, the fraction of water lost via fractionating routes was estimated from the isotopic abundances of body water, local drinking water, and dietary solids. Fractionated water loss averaged 23% (SD = 10%) of water turnover, which agreed with our previous estimates based on metabolic rate, but there was a systematic difference between the results based on O/sub 2/ and hydrogen. Corrections for isotopic fractionation of water lost in breath and (nonsweat) transcutaneous loss should be made when using labeled water to measure water turnover or CO/sub 2/ production.

  8. The role of technology in the past and future development of the doubly labelled water method.

    PubMed

    Speakman, John R

    2005-12-01

    The doubly labelled water method is an isotope-based technique that is used to measure the energy demands of free-living animals and humans. It is based on the observation that, in the body, the oxygen in carbon dioxide is in complete isotope exchange equilibrium with the oxygen in body water. Hence, a label of isotopic oxygen in body water is eliminated by both respiratory CO(2) and water turnover, whereas a similarly introduced label of deuterium is eliminated only by water flux. The difference in isotope fluxes therefore permits estimation of CO(2) production, which is correlated to energy demands. The doubly labelled water method has been advanced predominantly by technological advances in mass spectrometry. Although it was first described in the 1950s, it was only used on small animals and in low numbers because the costs of the isotopes were a primary constraint. However, advances in mass spectrometry precision and accuracy in the 1980s made it possible to reduce the quantities of isotope used, and hence apply the method on humans, although still in small numbers. The advent of continuous flow inlets in the 1990s made possible the processing of samples in much larger numbers and the sample sizes of studies have expanded. Ironically, however, the technique is now under treat because of technological advances in another area (positron emission tomography), which has generated an enormous demand for (18)O and pushed up the price of isotopes. A continuation of this trend might drive prices to levels where sustained application of the method in human studies is questionable. Replacing determination of isotope enrichments currently performed by isotope ratio mass spectrometry with determinations made by stable isotope infrared laser spectrometry may be a technological advance that will get us out of this problem.

  9. Energy expenditure in space flight (doubly labelled water method) (8-IML-1)

    NASA Technical Reports Server (NTRS)

    Parsons, Howard G.

    1992-01-01

    The objective of the Energy Expenditure in Space Flight (ESS) experiment is to demonstrate and evaluate the doubly labeled water method of measuring the energy expended by crew members during approximately 7 days in microgravity. The doubly labeled water technique determines carbon dioxide production which is then used to calculate energy expenditure. The method relies on the equilibrium between oxygen in respiratory carbon dioxide and oxygen in body water. Because of this equilibrium, the kinetic of water turnover and respiration are interdependent. Under normal conditions, man contains small but significant amounts of deuterium and oxygen 18. Deuterium is eliminated from the body as water while oxygen 18 is eliminated as water and carbon dioxide. The difference in the turnover rates in the two isotopes is proportional to the carbon dioxide production. Deliberately enriching the total body water with both of these isotopes allows the isotope turnovers to be accurately measured in urine, plasma, or saliva samples. The samples are taken to the laboratory for analysis using an ion-ratio spectrometer.

  10. Energy expenditure in space flight (doubly labelled water method) (8-IML-1)

    NASA Technical Reports Server (NTRS)

    Parsons, Howard G.

    1992-01-01

    The objective of the Energy Expenditure in Space Flight (ESS) experiment is to demonstrate and evaluate the doubly labeled water method of measuring the energy expended by crew members during approximately 7 days in microgravity. The doubly labeled water technique determines carbon dioxide production which is then used to calculate energy expenditure. The method relies on the equilibrium between oxygen in respiratory carbon dioxide and oxygen in body water. Because of this equilibrium, the kinetic of water turnover and respiration are interdependent. Under normal conditions, man contains small but significant amounts of deuterium and oxygen 18. Deuterium is eliminated from the body as water while oxygen 18 is eliminated as water and carbon dioxide. The difference in the turnover rates in the two isotopes is proportional to the carbon dioxide production. Deliberately enriching the total body water with both of these isotopes allows the isotope turnovers to be accurately measured in urine, plasma, or saliva samples. The samples are taken to the laboratory for analysis using an ion-ratio spectrometer.

  11. Natural abundance deuterium and 18-oxygen effects on the precision of the doubly labeled water method

    NASA Technical Reports Server (NTRS)

    Horvitz, M. A.; Schoeller, D. A.

    2001-01-01

    The doubly labeled water method for measuring total energy expenditure is subject to error from natural variations in the background 2H and 18O in body water. There is disagreement as to whether the variations in background abundances of the two stable isotopes covary and what relative doses of 2H and 18O minimize the impact of variation on the precision of the method. We have performed two studies to investigate the amount and covariance of the background variations. These were a study of urine collected weekly from eight subjects who remained in the Madison, WI locale for 6 wk and frequent urine samples from 14 subjects during round-trip travel to a locale > or = 500 miles from Madison, WI. Background variation in excess of analytical error was detected in six of the eight nontravelers, and covariance was demonstrated in four subjects. Background variation was detected in all 14 travelers, and covariance was demonstrated in 11 subjects. The median slopes of the regression lines of delta2H vs. delta18O were 6 and 7, respectively. Modeling indicated that 2H and 18O doses yielding a 6:1 ratio of final enrichments should minimize this error introduced to the doubly labeled water method.

  12. Natural abundance deuterium and 18-oxygen effects on the precision of the doubly labeled water method

    NASA Technical Reports Server (NTRS)

    Horvitz, M. A.; Schoeller, D. A.

    2001-01-01

    The doubly labeled water method for measuring total energy expenditure is subject to error from natural variations in the background 2H and 18O in body water. There is disagreement as to whether the variations in background abundances of the two stable isotopes covary and what relative doses of 2H and 18O minimize the impact of variation on the precision of the method. We have performed two studies to investigate the amount and covariance of the background variations. These were a study of urine collected weekly from eight subjects who remained in the Madison, WI locale for 6 wk and frequent urine samples from 14 subjects during round-trip travel to a locale > or = 500 miles from Madison, WI. Background variation in excess of analytical error was detected in six of the eight nontravelers, and covariance was demonstrated in four subjects. Background variation was detected in all 14 travelers, and covariance was demonstrated in 11 subjects. The median slopes of the regression lines of delta2H vs. delta18O were 6 and 7, respectively. Modeling indicated that 2H and 18O doses yielding a 6:1 ratio of final enrichments should minimize this error introduced to the doubly labeled water method.

  13. Validation of doubly labeled water method for energy expenditure in postsurgical infants

    SciTech Connect

    Jones, P.J.H.; Winthrop, A.L.; Schoeller, D.A.; Swyer, P.R.; Filler, R.M.; Smith, J.M.; Heim, T.

    1986-03-05

    To validate the doubly labeled water method (/sup 2/H/sub 2/ /sup 18/O) in infants without concurrent water balance, carbon dioxide production rate (rCO/sub 2/) and energy expenditure (EE) were measured for 5 or 6 days by /sup 2/H/sub 2/ /sup 18/O and periodic open circuit respiratory gas exchange in 5 infants (mean age: 5.3 wk, range 1-14 wk). Following abdominal surgery (mean = 10.3 d, range 7-18 d), infants were maintained on constant oral or parenteral nutrition 4 d prior to and during the study. This avoided changes in baseline isotopic enrichment of body water diet induced changes in relative isotopic abundance of /sup 2/H and /sup 18/O could introduce significant errors in rCO/sub 2/. For /sup 2/H/sub 2/ /sub 1//sup 8/O, they assumed insensible water loss would be proportional to respiration volume and body surface area and hence rCO/sub 2/. This calculated insensible loss averaged 18% of water turnover. EE was calculated using measured respiratory quotients (m) and dietary intake (i) data. In 6 blinded studies with 5 infants, rCO/sub 2/ = 34.2 L/d (range 27.3-48.0), EE = 191 kcal/d (133-266) and EE/sub i/ = 197 kcal/d (138-281). Percent differences (+/- SD) from respiratory gas exchange were -1.1 +/- 5.8, -1.1 +/- 5.8, and 1.4 +/- 5.2, respectively. These findings demonstrate the validity of the doubly labeled water method for determining energy expenditure without concurrent water balance studies.

  14. A rapid analytical technique for the determination of energy expenditure by the doubly labelled water method.

    PubMed

    Barrie, A; Coward, W A

    1985-09-01

    The doubly labelled water method involves the administration of water enriched in 2H and 18O followed by determination of the turnover rates of these isotopes. Since 18O is eliminated from the body as both CO2 and water, while 2H leaves only as water, the difference between the two turnover rates provides a measure of CO2 production and hence energy expenditure. Isotopic analysis by conventional stable isotope ratio analysis (SIRA) is labour intensive and time consuming, as it requires off-line conversion of water samples to gases (H2 and CO2) followed by sequential analysis for each of the two isotopes using the mass spectrometer. Lack of suitable automated instrumentation with the ability to process large numbers of samples has prevented routine application of the method. We describe here an automated technique in which body water samples (urine, saliva, breath water or milk) are analysed simultaneously for 2H and 18O. The single bench system comprises two mass spectrometer analysers, one for measuring 2H from H2 gas, the other for measuring 18O from the water vapour (masses 18, 20). Both analysers share a common heated inlet system into which microlitre quantities of the body fluids are injected from an autosampler (102 samples). The water vapour flows both directly to one analyser for 18O measurement and into a uranium reduction furnace for conversion to H2, prior to 2H measurement by the second analyser. Both analysers also share vacuum and electronic components, enabling savings in both space and cost. In this paper we present results illustrating performance characteristics and procedures for routine application to human subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. Blood-sucking bugs as a gentle method for blood-collection in water budget studies using doubly labelled water.

    PubMed

    Voigt, Christian C; Michener, Robert; Wibbelt, Gudrun; Kunz, Thomas H; von Helversen, Otto

    2005-11-01

    During doubly-labelled water (DLW) experiments, blood collection by venous puncture may traumatize animals and consequently affect the animals' behaviour and energy budget. Recent studies have shown that blood-sucking bugs (Triatominae; Heteroptera) can be used instead of conventional needles to obtain blood from animals. In this paper, we validate the bug method in captive nectar-feeding bats, Glossophaga soricina, for water budget analysis by comparing the daily water flux estimated with the DLW method with values measured by an energy balance method. As the mean daily water flux of the DLW method was not significantly deviating from the expected value, blood-sucking bugs may substitute more invasive methods of blood collection in DLW experiments. Based on the DLW estimates, daily energy and water intake rates were calculated and compared to values measured with the energy balance method. The DLW method and the energy balance method yielded on average similar results regarding the daily energy intake (DLW method: 48.8+/-14.2 kJ d(-1) versus energy balance method: 48.1+/-9.9 kJ d(-1)) and daily water intake (DLW method: 13.7+/-2.4 mL d(-1) versus energy balance method: 14.7+/-3.0 mL d(-1)). Based on the calculated water and sugar intake per day, we estimated the sugar concentration of ingested nectar to equal on average 16.2+/-2.4% (mass/mass), which fell close to the measured sugar concentration of 17% (mass/mass) bats fed on during the experiment. We conclude that it is possible to extrapolate mean daily energy and water intake for animal groups, populations and species based on DLW estimates, but due to the large variance of results (low accuracy), it seems inadequate to calculate values for single individuals.

  16. Contributions of the doubly labeled water method to studies of energy balance in the Third World.

    PubMed

    Coward, W A

    1998-10-01

    Of >250 studies on energy metabolism using the doubly labeled water (DLW) technique, approximately 12 full papers describe work performed in the Third World. Unfortunately, the term "Third World" is imprecise and the focuses of individual studies were too varied to allow much comparison among the data. There is a need to develop a more uniform approach. Useful investigations will allow comparisons of energy metabolism to be made in a consistent way across a variety of socioeconomic groups within the same country, and between the Third World and the developed world, with a commitment to the long term. In this way, the DLW method, if combined with other measurements of activity, energy intake, and body composition, will provide useful information on energy requirements and the consequences of inadequate or excessive energy intakes for the individual. Such investigations should be done, with standard protocols where possible, not just for the DLW method but also for those methods with which DLW should be integrated. Only if these suggestions are followed will real "value for money" be obtained from DLW studies in the Third World or elsewhere. In the context of these criteria, studies in the Third World using the DLW method have been only partly successful.

  17. A review of seabird energetics using the doubly labeled water method.

    PubMed

    Shaffer, Scott A

    2011-03-01

    The doubly labeled water (DLW) method has been essential for understanding animal energetics of free-ranging individuals. The first published studies on free-ranging seabirds were conducted on penguins in the early 1980s. Since then, nearly 50 seabird species with representatives from each major taxonomic order have been studied using DLW. Although the basic methodology has not changed, there are at least nine different equations, varying with respect to assumptions on fractionation and the total body water pool, to estimate field metabolic rate (FMR) from isotopic water turnover. In this review, I show that FMR can vary by as much as 45% depending on the equation used to calculate CO(2) production in five albatross species. Energy budgets derived from DLW measurements are critical tools for understanding patterns of energy use and allocation in seabirds. However, they depend on accurate and representative measurements of FMR, so analyses that include greater partitioning of activity specific FMR yield more realistic cost estimates. I also show how the combined use of DLW and biologging methods can 1) provide greater clarity for explaining observed variation in FMR measurements within a species and 2) allow FMRs to be viewed in a wider physiological, behavioral, or ecological context. Finally, I update existing allometric equations with new FMR data. These updates reaffirm that albatrosses have the lowest at-sea FMRs per equivalent body mass and that individuals of other seabird orders have FMRs ranging between 1.39 and 2.24 times higher than albatrosses. Copyright © 2010 Elsevier Inc. All rights reserved.

  18. Energy Requirement Assessment in Japanese Table Tennis Players Using the Doubly Labeled Water Method.

    PubMed

    Sagayama, Hiroyuki; Hamaguchi, Genki; Toguchi, Makiko; Ichikawa, Mamiko; Yamada, Yosuke; Ebine, Naoyuki; Higaki, Yasuki; Tanaka, Hiroaki

    2017-05-22

    Total daily energy expenditure (TEE) and physical activity level (PAL) are important for adequate nutritional management in athletes. The PAL of table tennis has been estimated to about 2.0: it is categorized as a moderate-activity sport (4.0 metabolic equivalents [METs]) in the Compendium of Physical Activities. However, modern table tennis makes high physiological demands. The aims of the present study were to examine (1) TEE and PAL of competitive table tennis players and (2) the physiological demands of various types of table tennis practice. In Experiment 1, we measured TEE and PAL in 10 Japanese college competitive table tennis players (aged 19.9 ± 1.1 years) using the doubly labeled water (DLW) method during training and with an exercise training log and self-reported energy intake. TEE was 15.5 ± 1.9 MJ·day(-1) (3695 ± 449 kcal·day(-1)); PAL was 2.53 ± 0.25; and the average training duration was 181 ± 38 min·day(-1). In Experiment 2, we measured METs of five different practices in seven college competition players (20.6 ± 1.2 years). Three practices without footwork were 4.5-5.2 METs, and two practices with footwork were 9.5-11.5 METs. Table tennis practices averaged 7.1 ± 3.2 METS demonstrating similarities with other vigorous racket sports. In conclusion the current Compendium of Physical Activities underestimates the physiological demands of table tennis practice for competition; the estimated energy requirement should be based on DLW method data.

  19. Total energy expenditure by the doubly-labeled water method in rural preschool children in Cuba.

    PubMed

    Hernández-Triana, Manuel; Salazar, Gabriela; Díaz, Erik; Sánchez, Vivian; Basabe, Beatriz; González, Soraya; Díaz, Maria Elena

    2002-09-01

    An evaluation of the capacity of the Cuban Nutrition Program for covering the energy requirements of children was carried out in children 5.0 +/- 1.0 years of age in a rural mountain community in Cuba. Five males and six females (19 +/- 3.05 kg average weight) with a mean BMI 15.4 +/- 2.2 kg/m2) were included in the study. Six of the subjects were well-nourished, three were undernourished, and two were overweight. Total energy expenditure was determined by the doubly-labeled water technique. Resting metabolic rate was measured by indirect calorimetry. Energy intake, measured by a three-day weighed dietary record, was 1,527 kcal/day (6.39 MJ). The total energy expenditure of the well-nourished children was 11.8% lower than present energy recommendations (1,773 kcal). This implies that well-nourished children who are moderately to heavily physically active require 82.6 kcal/kg per day. The physical activity level of normal Cuban children is nearly 1.8, which is much higher than that reported in studies of children from industrialized countries using the doubly-labeled water technique. The measured daily energy intake was 1.7 times the resting metabolic rate RMR and 1.04 times the total energy expenditure.

  20. Effect of bolus fluid intake on energy expenditure values as determined by the doubly labeled water method

    NASA Technical Reports Server (NTRS)

    Drews, D.; Stein, T. P.

    1992-01-01

    The doubly labeled water (DLW, 2H(2)18O) method is a highly accurate method for measuring energy expenditure (EE). A possible source of error is bolus fluid intake before body water sampling. If there is bolus fluid intake immediately before body water sampling, the saliva may reflect the ingested water disproportionately, because the ingested water may not have had time to mix fully with the body water pool. To ascertain the magnitude of this problem, EE was measured over a 5-day period by the DLW method. Six subjects were dosed with 2H2(18)O. After the reference salivas for the two-point determination were obtained, subjects drank water (700-1,000 ml), and serial saliva samples were collected for the next 3 h. Expressing the postbolus saliva enrichments as a percentage of the prebolus value, we found 1) a minimum in the saliva isotopic enrichments were reached at approximately 30 min with the minimum for 2H (95.48 +/- 0.43%) being significantly lower than the minimum for 18O (97.55 +/- 0.44, P less than 0.05) and 2) EE values calculated using the postbolus isotopic enrichments are appreciably higher (19.9 +/- 7.5%) than the prebolus reference values. In conclusion, it is not advisable to collect saliva samples for DLW measurements within approximately 1 h of bolus fluid intake.

  1. Effect of bolus fluid intake on energy expenditure values as determined by the doubly labeled water method

    NASA Technical Reports Server (NTRS)

    Drews, D.; Stein, T. P.

    1992-01-01

    The doubly labeled water (DLW, 2H(2)18O) method is a highly accurate method for measuring energy expenditure (EE). A possible source of error is bolus fluid intake before body water sampling. If there is bolus fluid intake immediately before body water sampling, the saliva may reflect the ingested water disproportionately, because the ingested water may not have had time to mix fully with the body water pool. To ascertain the magnitude of this problem, EE was measured over a 5-day period by the DLW method. Six subjects were dosed with 2H2(18)O. After the reference salivas for the two-point determination were obtained, subjects drank water (700-1,000 ml), and serial saliva samples were collected for the next 3 h. Expressing the postbolus saliva enrichments as a percentage of the prebolus value, we found 1) a minimum in the saliva isotopic enrichments were reached at approximately 30 min with the minimum for 2H (95.48 +/- 0.43%) being significantly lower than the minimum for 18O (97.55 +/- 0.44, P less than 0.05) and 2) EE values calculated using the postbolus isotopic enrichments are appreciably higher (19.9 +/- 7.5%) than the prebolus reference values. In conclusion, it is not advisable to collect saliva samples for DLW measurements within approximately 1 h of bolus fluid intake.

  2. Estimating free-living human energy expenditure: Practical aspects of the doubly labeled water method and its applications

    PubMed Central

    Kazuko, Ishikawa-Takata; Kim, Eunkyung; Kim, Jeonghyun; Yoon, Jinsook

    2014-01-01

    The accuracy and noninvasive nature of the doubly labeled water (DLW) method makes it ideal for the study of human energy metabolism in free-living conditions. However, the DLW method is not always practical in many developing and Asian countries because of the high costs of isotopes and equipment for isotope analysis as well as the expertise required for analysis. This review provides information about the theoretical background and practical aspects of the DLW method, including optimal dose, basic protocols of two- and multiple-point approaches, experimental procedures, and isotopic analysis. We also introduce applications of DLW data, such as determining the equations of estimated energy requirement and validation studies of energy intake. PMID:24944767

  3. WaterSense Labeled New Homes

    EPA Pesticide Factsheets

    Homes built to meet EPA’s specification can earn the WaterSense label. EPA criteria include WaterSense labeled plumbing fixtures, efficient hot water delivery systems, water-smart landscape design, and other features.

  4. Validity of the Modified Baecke Questionnaire: comparison with energy expenditure according to the doubly labeled water method

    PubMed Central

    Hertogh, Emmy M; Monninkhof, Evelyn M; Schouten, Evert G; Peeters, Petra HM; Schuit, Albertine J

    2008-01-01

    Background In epidemiological research, physical activity is usually assessed by questionnaires. Questionnaires are suitable for large study populations since they are relatively inexpensive and not very time consuming. However, questionnaire information is by definition subjective and prone to recall bias, especially among elderly subjects. The Modified Baecke Questionnaire, developed by Voorrips and coworkers, measures habitual physical activity in the elderly. The questionnaire includes questions on household activities, sports, and leisure time activities, over a time period of one year. The Modified Baecke Questionnaire results in a score to classify people as high, moderate, or low in daily physical activity, based on tertiles. Methods The validity of the Modified Baecke Questionnaire score was assessed among 21 elderly men and women using the doubly labeled water method as the reference criterion. This method is considered to be the gold standard for measuring energy expenditure in free-living individuals. Energy expenditure on physical activity is estimated by the ratio of total energy expenditure measured by the doubly labeled water method and resting metabolic rate measured by indirect calorimetry. This ratio is called the physical activity ratio. Results The Spearman correlation coefficient between the questionnaire score and the physical activity ratio (PAR) was 0.54 (95% CI 0.22–0.66). Correct classification by the questionnaire occurred in 71% of participants who were in the lowest tertile of PAR, in 14% of participants in the middle tertile, and in 43% of participants in the highest tertile. Subjects were not wrongly classified in an opposite tertile. Conclusion The validity of the Modified Baecke Questionnaire is fair-to-moderate. This study shows that the questionnaire can correctly classify individuals as low or high active, but does a poor job for moderately active individuals. PMID:18505554

  5. Effect of weaning on accuracy of doubly labeled water method in infants

    SciTech Connect

    Roberts, S.B.; Coward, W.A.; Ewing, G.; Savage, J.; Cole, T.J.; Lucas, A.

    1988-04-01

    Variations in background /sup 2/H and /sup 18/O abundances in body water influence the accuracy and precision of the /sup 2/H/sub 2//sup 18/O method for determination of energy expenditure. To investigate the effect of weaning during infancy on background /sup 2/H and /sup 18/O abundances, urine samples from 44 breast- or formula-fed infants aged 5-16 wk were analyzed. /sup 2/H and /sup 18/O abundances were significantly higher (P less than 0.001) in breast- than in formula-fed infants. The relationship between /sup 2/H and /sup 18/O abundances was linear and independent of diet (slope, 4.16 +/- 0.43 (SE)). By use of this information, the effect of weaning on the accuracy of the /sup 2/H/sub 2//sup 18/O method was evaluated, taking into account the effect of /sup 2/H-/sup 18/O abundances in the isotope loading dose. In infants weaned completely from breast milk to formula during the measurement, energy expenditure can be overestimated by 18.0%, even if /sup 2/H-/sup 18/O abundances in the isotope dose equal the ratio of naturally occurring background changes. However, this error can be reduced to less than 3.0% by manipulating the study duration and isotope dose. During gradual weaning, the overestimation of energy expenditure is only 0.3-2.0%.

  6. Validation of the doubly labeled water method using off-axis integrated cavity output spectroscopy and isotope ratio mass spectrometry.

    PubMed

    Melanson, Edward L; Swibas, Tracy; Kohrt, Wendy M; Catenacci, Vicki A; Creasy, Seth A; Plasqui, Guy; Wouters, Loek; Speakman, John R; Berman, Elena S F

    2017-10-03

    When the doubly-labeled water (DLW) method is used to measure total daily energy expenditure (TDEE), isotope measurements are typically performed using isotope ratio mass spectrometry (IRMS). New technologies, such as off-axis integrated cavity output spectroscopy (OA-ICOS) provide comparable isotopic measurements of standard waters and human urine samples, but the accuracy of carbon dioxide production (VCO2) determined with OA-ICOS has not been demonstrated. We compared simultaneous measurement of VCO2 obtained using whole-room indirect calorimetry (IC) with DLW-based measurements from IRMS and OA-ICOS. 17 subjects (10 female; 22 to 63 yrs.) were studied for 7 consecutive days in the IC. Subjects consumed a dose of 0.25 g H2(18)O (95% APE) and 0.14 g (2)H2O (99.8% APE) per kg of total body water, and urine samples were obtained on days 1 and 8 to measure average daily VCO2 using OA-ICOS and IRMS. VCO2 was calculated using both the plateau and intercept methods. There were no differences in VCO2 or TDEE measured by OA-ICOS or IRMS compared with IC when the plateau method was used. When the intercept method was used, VCO2 measured using OA-ICOS did not differ from IC, but VCO2 measured using IRMS was significantly lower than IC. Accuracy (~1-5%), precision (~8%), intraclass correlation coefficients (R=0.87-90), and root mean squared error (30-40 L/day) of VCO22 measured by OA-ICOS and IRMS were similar. Both OA-ICOS and IRMS produced measurements of VCO2 with comparable accuracy and precision when compared to IC. Copyright © 2017, American Journal of Physiology-Endocrinology and Metabolism.

  7. Validation of the doubly-labeled water (H/sup 3/H/sup 18/O) method for measuring water flux and energy metabolism in tenebrionid beetles

    SciTech Connect

    Cooper, P.D.

    1981-01-01

    Doubly-labeled water (H/sup 3/H/sup 18/O) has been used to determine water flux and energy metabolism in a variety of vertebrates. This study examines the applicability of this technique to arthropods. The theory of the technique depends upon the assumption that doubly-labeled water introduced into the animal's body water equilibrates with water and carbon dioxide by the action of carbonic anhydrase. Tritium (/sup 3/H) is lost from the animal only with water while oxygen-18 is lost with both water and carbon dioxide. The difference bwtween the rates of loss of the two isotopes is proportional to CO/sub 2/ loss rate. Validation of the use of tritiated water for measuring water flux was accomplished by comparing gravimetric measurements of water gain with flux rates determined by loss of tritiated water. At room humidity, an overestimate for influx calculated from labeled water calculations was found, averaging 12 mg H/sub 2/O (g.d)/sup -1/. Comparison of CO/sub 2/ loss rate determined isotopically with rates of CO/sub 2/ loss determined by standard metabolic rates also yielded overestimates for the isotopic technique, overestimates ranging between 20 and 30%. The relevance of this for studies using labeled water for studying water fluxes and free metabolism of free-ranging arthropods is discussed.

  8. Simultaneous measurement of free-living energy expenditure by the doubly labeled water method and heart-rate monitoring

    SciTech Connect

    Livingstone, M.B.; Prentice, A.M.; Coward, W.A.; Ceesay, S.M.; Strain, J.J.; McKenna, P.G.; Nevin, G.B.; Barker, M.E.; Hickey, R.J. )

    1990-07-01

    Total energy expenditure (TEE) was measured simultaneously in 14 free-living adults over 15 d by the doubly labeled water (DLW) method and for 2-4 separate days by heart-rate (HR) monitoring. Individual curves for HR vs oxygen consumption (VO2) were obtained and an HR (FLEX HR: 97 +/- 8 beats/min, range 84-113 beats/min) that discriminated between rest and activity was identified. Calibration curves were used to assign an energy value to daytime HR above FLEX HR. Below FLEX HR energy expenditure was taken as resting metabolism. Sleeping energy expenditure was assumed to be equal to basal metabolic rate. Average HR TEE (12.99 +/- 3.83 MJ/d) and average DLW TEE (12.89 +/- 3.80 MJ/d) were similar. HR TEE discrepancies ranged from -22.2% to +52.1%, with nine values lying within +/- 10% of DLW TEE estimates. The FLEX HR method provides a close estimation of the TEE of population groups. However, an increased number of sampling days may improve the precision of individual estimates of TEE.

  9. Assessment of energy requirements in patients with short bowel syndrome by using the doubly labeled water method.

    PubMed

    Fassini, Priscila Giacomo; Pfrimer, Karina; Ferriolli, Eduardo; Suen, Vivian Miguel Marques; Marchini, Júlio Sérgio; Das, Sai Krupa

    2016-01-01

    Short bowel syndrome (SBS) is a serious malabsorption disorder, and dietetic management of patients with SBS is extremely challenging. Once the degree of undernutrition has been assessed, successful dietary intervention is contingent on an accurate estimation and provision of energy needs. We quantified total energy expenditure (TEE) in patients with SBS by using the doubly labeled water (DLW) method to inform energy needs and nutritional therapy goals. In this observational study, TEE was measured in 22 participants, 11 with SBS and 11 sex-, age-, and body mass index (BMI)-matched controls (non-SBS), for 14 d with the DLW method. Predicted energy requirements were determined by using the Escott-Stump equation and compared with TEE determined with DLW. Resting energy expenditure was measured by using indirect calorimetry, and an accelerometer was also used to determine physical activity level. Participants were aged (mean ± SD) 53 ± 8 y. Measured TEE was significantly higher than predicted TEE for the SBS group (1875 ± 276 compared with 1517 ± 175 kcal/d, P = 0.001) and also for the non-SBS group (2393 ± 445 compared with 1532 ± 178 kcal/d, P < 0.01). Measured TEE was significantly lower in the SBS group than in the non-SBS group (P < 0.01); however, predicted TEE did not differ significantly between the groups (P = 0.84). No significant differences were seen between measured and predicted resting energy expenditure either within or between groups. Measured TEE in patients with SBS was significantly higher than predicted by using standard equations but also lower than values for age-, BMI-, and sex-matched non-SBS controls. Currently used formulas in clinical practice appear to underestimate energy requirements of patients with SBS, and revision is needed to prevent underfeeding and improve long-term prognosis. This trial was registered at clinicaltrials.gov as NCT02113228. © 2016 American Society for Nutrition.

  10. Energy expenditure of Chinese infants in Guangdong Province, south China, determined with use of the doubly labeled water method.

    PubMed

    Jiang, Z; Yan, Q; Su, Y; Acheson, K J; Thélin, A; Piguet-Welsch, C; Ritz, P; Ho, Z C

    1998-06-01

    The doubly labeled water method was used to measure the energy expenditure of a group of 41, 4- or 6-mo-old infants with a cross-sectional design. The infants were divided into two groups according to whether they were breast-fed (11 at 4 mo, 9 at 6 mo) or formula fed (11 at 4 mo, 10 at 6 mo). Anthropometric measurements were recorded at birth and at the beginning and end of the 8-d study. Anthropometric data, which were supported by the food intake and energy expenditure results, indicated that the infants were within the norms for European and American infants of the same age. Mean energy intakes of 352 kJ (84 kcal) kg(-1) x d(-1) at 4 and 6 mo were lower than the FAO/WHO/UNU recommended value of 452 kJ (108 kcal) x kg(-1) x d(-1)) and Chinese recommendations of 502 kJ (120 kcal) kg(-1) x d(-1). However, some authors believe that values of 398 kJ (95 kcal) kg(-1) x d(-1) at 4 mo and 356 kJ (85 kcal) kg(-1) x d(-1) at 6 mo are more appropriate. At 6 mo the infants' length-for-age and weight-for-age were at the National Center for Health Statistics 55th and 47th percentiles, respectively, whereas 58% were below the 50th percentile for weight-for-length. We conclude that at 4 mo infants receive sufficient energy for their requirements. However, at 6 mo energy requirements might well be greater than the revised recommendations, when infants are being weaned to alternative foods and are more prone to the influence of diet on their growth and development.

  11. Determination of energy expenditure during heavy exercise, normal daily activity, and sleep using the doubly-labelled-water (/sup 2/H/sub 2/ 18O) method

    SciTech Connect

    Stein, T.P.; Hoyt, R.W.; Settle, R.G.; O'Toole, M.; Hiller, W.D.

    1987-03-01

    Energy expenditure of four subjects was measured by the doubly-labelled-water (/sup 2/H/sub 2/ 18O) method to determine if energy expenditure could be determined over short periods. Three subjects were studied while they performed 8 h of heavy exercise in a laboratory environment. Urine and blood samples were taken before and after exercise. Estimated energy expended during 8 h of high-intensity exercise for three subjects was 757 +/- 118 kcal/h by the doubly-labelled-water method using urine and a two-point calculation, which compared favorably with 735 +/- 82 kcal/h obtained by respiratory gas exchange. For the fourth subject, daytime, nighttime, and daily energy expenditure was calculated by both the two-pair method and decay-curve analysis of urine and saliva samples collected in the morning and at night. Daytime and nighttime energy expenditures differed significantly (p less than 0.05).

  12. Pre-embedding labeling methods.

    PubMed

    Oliver, Constance

    2010-01-01

    Colloidal gold conjugates generally do not readily penetrate cells, even after permeabilization. Therefore, their use in pre-embedding immunostaining has been largely restricted to labeling cell-surface antigens for scanning or transmission electron microscopy or for tracing endocytic pathways in living cells. One nanometer gold conjugates that do penetrate cells and tissues much more readily have also been used successfully to immunolabel intracellular structures. For pre-embedding labeling, all of the immunostaining is done prior to embedding the tissue in resin or preparing the samples for scanning electron microscopy. This chapter provides methods for pre-embedding staining with unconjugated primary antibody or with primary antibody conjugated to colloidal gold. The use of colloidal gold for tracing endocytic pathways is also given.

  13. Peroral administration of 5-bromo-2-deoxyuridine in drinking water is not a reliable method for labeling proliferating S-phase cells in rats.

    PubMed

    Ševc, Juraj; Matiašová, Anna; Smoleková, Ivana; Jendželovský, Rastislav; Mikeš, Jaromír; Tomášová, Lenka; Kútna, Viera; Daxnerová, Zuzana; Fedoročko, Peter

    2015-01-01

    In rodents, peroral (p.o.) administration of 5-bromo-2'-deoxyuridine (BrdU) dissolved in drinking water is a widely used method for labeling newly formed cells over a prolonged time-period. Despite the broad applicability of this method, the pharmacokinetics of BrdU in rats or mice after p.o. administration remains unknown. Moreover, the p.o. route of administration may be limited by the relatively low amount of BrdU consumed over 24h and the characteristic drinking pattern of rats, with water intake being observed predominantly during the dark phase. Therefore, we investigated the reliability of staining proliferating S-phase cells with BrdU after p.o. administration (1mg/ml) to rats using both in vitro and in vivo conditions. Flow cytometric analysis of tumor cells co-cultivated with sera from experimental animals exposed to BrdU dissolved in drinking water or 25% orange juice revealed that the concentration of BrdU in the blood sera of rats throughout the day was below the detection limits of our assay. Ingested BrdU was only sufficient to label approximately 4.2±0.3% (water) or 4.2±0.3% (25% juice) of all S-phase cells. Analysis of data from in vivo conditions indicates that only 7.6±3.3% or 15.5±2.3% of all S-phase cells in the dentate gyrus of the hippocampus was labeled in animals administered drinking water containing BrdU during the light and dark phases of the day. In addition, the intensity of BrdU-positive nuclei in animals receiving p.o. administration of BrdU was significantly lower than in control animals intraperitoneally injected with BrdU. Our data indicate that the conventional approach of p.o. administration of BrdU in the drinking water to rats provides strongly inaccurate information about the number of proliferating cells in target tissues. Therefore other administration routes, such as osmotic mini pumps, should be considered for labeling of proliferating cells over a prolonged time-period. Copyright © 2015 Elsevier Inc. All rights

  14. Apparatus and method for preparing oxygen-15 labeled water H{sub 2}[{sup 15}O] in an injectable form for use in positron emission tomography

    DOEpatents

    Ferrieri, R.A.; Schlyer, D.J.; Alexoff, D.

    1996-01-09

    A handling and processing apparatus is revealed for preparing Oxygen-15 labeled water (H{sub 2}[{sup 15}O]) in injectable form for use in Positron Emission Tomography from preferably H{sub 2}[{sup 15}O] produced by irradiating a flowing gas target of nitrogen and hydrogen. The apparatus includes a collector for receiving and directing a gas containing H{sub 2}[{sup 15}O] gas and impurities, mainly ammonia (NH{sub 3}) gas into sterile water to trap the H{sub 2}[{sup 15}O] and form ammonium (NH{sub 4}{sup +}) in the sterile water. A device for displacing the sterile water containing H{sub 2}[{sup 15}O] and NH{sub 4}{sup +} through a cation resin removes NH{sub 4}{sup +} from the sterile water. A device for combining the sterile water containing H{sub 2}[{sup 15}O] with a saline solution produces an injectable solution. Preferably, the apparatus includes a device for delivering the solution to a syringe for injection into a patient. Also, disclosed is a method for preparing H{sub 2}[{sup 15}O] in injectable form for use in Positron Emission Tomography in which the method neither requires isotopic exchange reaction nor application of high temperature. 7 figs.

  15. Apparatus and method for preparing oxygen-15 labeled water H.sub.2 [.sup.15 O] in an injectable form for use in positron emission tomography

    DOEpatents

    Ferrieri, Richard A.; Schlyer, David J.; Alexoff, David

    1996-01-09

    A handling and processing apparatus for preparing Oxygen-15 labeled water (H.sub.2 [.sup.15 O]) in injectable form for use in Positron Emission Tomography from preferably H.sub.2 [.sup.15 O] produced by irradiating a flowing gas target of nitrogen and hydrogen. The apparatus includes a collector for receiving and directing a gas containing H.sub.2 [.sup.15 O] gas and impurities, mainly ammonia (NH.sub.3) gas into sterile water to trap the H.sub.2 [.sup.15 O] and form ammonium (NH.sub.4.sup.+) in the sterile water. A device for displacing the sterile water containing H.sub.2 [.sup.15 O] and NH.sub.4.sup.+ through a cation resin removes NH.sub.4.sup.+ from the sterile water. A device for combining the sterile water containing H.sub.2 [.sup.15 O] with a saline solution produces an injectable solution. Preferably, the apparatus includes a device for delivering the solution to a syringe for injection into a patient. Also, disclosed is a method for preparing H.sub.2 [.sup.15 O] in injectable form for use in Positron Emission Tomography in which the method neither requires isotopic exchange reaction nor application of high temperature.

  16. [Study on the validation of the computer science application's activity monitor in assessing the physical activity among adults using doubly labeled water method].

    PubMed

    Liu, Ai-ling; Li, Yan-ping; Song, Jun; Pan, Hui; Han, Xiu-ming; Ma, Guan-sheng

    2005-03-01

    Using doubly labeled water method to validate the colmputer science application's activity monitor (CSA) in assessing physical activity of free-living adults in Beijing, in order to develop equations to predict total daily energy expenditure (TEE) and activity related energy expenditure (AEE) from activity counts (AC) and anthropometric variables. A total of 72 healthy adults (33 males and 39 females, mean age 43.6 +/- 4.0 yr) were monitored for 7 consecutive days by CSA. TEE was simultaneously measured using doubly labeled water method. Average AC (counts/min(-1)) was compared with TEE, AEE and physical activity level (PAL). Physical activity determined by AC was significantly related to data on energy expenditures: TEE (r = 0.31, P < 0.01), AEE (r = 0.30, P < 0.05), and PAL (r = 0.26, P < 0.05). Multiple stepwise regression analysis showed that TEE was significantly influenced by gender, fat-free mass (FFM) or BMI and AC (R(2) = 0.52 - 0.70) while AEE was significantly influenced by gender, FFM and AC (R(2) = 0.25 - 0.32). AC from CSA activity monitor seemed a useful measure in studying the total amount of physical activity in free-living adults while AC significantly contributed to the explained variation in TEE and AEE.

  17. Two-point vs multipoint sample collection for the analysis of energy expenditure by use of the doubly labeled water method

    SciTech Connect

    Welle, S. )

    1990-12-01

    Energy expenditure over a 2-wk period was determined by the doubly labeled water (2H2(18)O) method in nine adults. When daily samples were analyzed, energy expenditure was 2859 +/- 453 kcal/d (means +/- SD); when only the first and last time points were considered, the mean calculated energy expenditure was not significantly different (2947 +/- 430 kcal/d). An analysis of theoretical cases in which isotope flux is not constant indicates that the multipoint method can cause errors in the calculation of average isotope fluxes, but these are generally small. Simulations of the effect of analytical error indicate that increasing the number of replicates on two points reduces the impact of technical errors more effectively than does performing single analyses on multiple samples. It appears that generally there is no advantage to collecting frequent samples when the 2H2(18)O method is used to estimate energy expenditure in adult humans.

  18. Efficient Methods for Multi-Label Classification

    DTIC Science & Technology

    2015-05-22

    annotation and retrieval of music and sound effects . IEEE Transactions on Audio, Speech and Language Processing 16(2), 467–476 (2008) 19. Vens, C., Struyf...advertising [2] and music categorization [18]. In these applications there are usually tens or hundreds of thousands of labels, while the number is...Efficient Methods for Multi-label Classiffication 165 and testing efficiency, memory usage is also a bottleneck as the number of labels becoming larger

  19. Accuracy of bottled drinking water label content.

    PubMed

    Khan, Nazeer B; Chohan, Arham N

    2010-07-01

    The purpose of the study was to compare the accuracy of the concentration of fluoride (F), calcium (Ca), pH, and total dissolved solids (TDS) levels mentioned on the labels of the various brands of bottled drinking water available in Riyadh, Saudi Arabia. Twenty-one different brands of locally produced non-carbonated (still water) bottled drinking water were collected from the supermarkets of Riyadh. The concentration of F, Ca, TDS, and pH values were noted from the labels of the bottles. The samples were analyzed for concentrations in the laboratory using the atomic absorption spectrophotometer. The mean level of F, Ca, and pH were found as 0.86 ppm, 38.47 ppm, and 7.5, respectively, which were significantly higher than the mean concentration of these elements reported in the labels. Whereas, the mean TDS concentration was found 118.87 ppm, which was significantly lower than the mean reported on the labels. In tropical countries like Saudi Arabia, the appropriate level of F concentration in drinking water as recommended by World Health Organization (WHO) should be 0.6-0.7 ppm. Since the level of F was found to be significantly higher than the WHO recommended level, the children exposed to this level could develop objectionable fluorosis. The other findings, like pH value, concentrations of Ca, and TDS, were in the range recommended by the WHO and Saudi standard limits and therefore should have no obvious significant health implications.

  20. Label-fracture: a method for high resolution labeling of cell surfaces.

    PubMed

    Pinto da Silva, P; Kan, F W

    1984-09-01

    We introduce here a technique, "label-fracture," that allows the observation of the distribution of a cytochemical label on a cell surface. Cell surfaces labeled with an electron-dense marker (colloidal gold) are freeze-fractured and the fracture faces are replicated by plantinum/carbon evaporation. The exoplasmic halves of the membrane, apparently stabilized by the deposition of the Pt/C replica, are washed in distilled water. The new method reveals the surface distribution of the label coincident with the Pt/C replica of the exoplasmic fracture face. Initial applications indicate high resolution (less than or equal to 15 nm) and exceedingly low background. "Label-fracture" provides extensive views of the distribution of the label on membrane surfaces while preserving cell shape and relating to the freeze-fracture morphology of exoplasmic fracture faces. The regionalization of wheat germ agglutinin receptors on the plasma membranes of boar sperm cells is illustrated. The method and the interpretation of its results are straightforward. Label-fracture is appropriate for routine use as a surface labeling technique.

  1. A Mobile Phone Based Method to Assess Energy and Food Intake in Young Children: A Validation Study against the Doubly Labelled Water Method and 24 h Dietary Recalls.

    PubMed

    Delisle Nyström, Christine; Forsum, Elisabet; Henriksson, Hanna; Trolle-Lagerros, Ylva; Larsson, Christel; Maddison, Ralph; Timpka, Toomas; Löf, Marie

    2016-01-15

    Mobile phones are becoming important instruments for assessing diet and energy intake. We developed the Tool for Energy Balance in Children (TECH), which uses a mobile phone to assess energy and food intake in pre-school children. The aims of this study were: (a) to compare energy intake (EI) using TECH with total energy expenditure (TEE) measured via doubly labelled water (DLW); and (b) to compare intakes of fruits, vegetables, fruit juice, sweetened beverages, candy, ice cream, and bakery products using TECH with intakes acquired by 24 h dietary recalls. Participants were 39 healthy, Swedish children (5.5 ± 0.5 years) within the ongoing Mobile-based Intervention Intended to Stop Obesity in Preschoolers (MINISTOP) obesity prevention trial. Energy and food intakes were assessed during four days using TECH and 24 h telephone dietary recalls. Mean EI (TECH) was not statistically different from TEE (DLW) (5820 ± 820 kJ/24 h and 6040 ± 680 kJ/24 h, respectively). No significant differences in the average food intakes using TECH and 24 h dietary recalls were found. All food intakes were correlated between TECH and the 24 h dietary recalls (ρ = 0.665-0.896, p < 0.001). In conclusion, TECH accurately estimated the average intakes of energy and selected foods and thus has the potential to be a useful tool for dietary studies in pre-school children, for example obesity prevention trials.

  2. A Mobile Phone Based Method to Assess Energy and Food Intake in Young Children: A Validation Study against the Doubly Labelled Water Method and 24 h Dietary Recalls

    PubMed Central

    Delisle Nyström, Christine; Forsum, Elisabet; Henriksson, Hanna; Trolle-Lagerros, Ylva; Larsson, Christel; Maddison, Ralph; Timpka, Toomas; Löf, Marie

    2016-01-01

    Mobile phones are becoming important instruments for assessing diet and energy intake. We developed the Tool for Energy Balance in Children (TECH), which uses a mobile phone to assess energy and food intake in pre-school children. The aims of this study were: (a) to compare energy intake (EI) using TECH with total energy expenditure (TEE) measured via doubly labelled water (DLW); and (b) to compare intakes of fruits, vegetables, fruit juice, sweetened beverages, candy, ice cream, and bakery products using TECH with intakes acquired by 24 h dietary recalls. Participants were 39 healthy, Swedish children (5.5 ± 0.5 years) within the ongoing Mobile-based Intervention Intended to Stop Obesity in Preschoolers (MINISTOP) obesity prevention trial. Energy and food intakes were assessed during four days using TECH and 24 h telephone dietary recalls. Mean EI (TECH) was not statistically different from TEE (DLW) (5820 ± 820 kJ/24 h and 6040 ± 680kJ/24 h, respectively). No significant differences in the average food intakes using TECH and 24 h dietary recalls were found. All food intakes were correlated between TECH and the 24 h dietary recalls (ρ = 0.665–0.896, p < 0.001). In conclusion, TECH accurately estimated the average intakes of energy and selected foods and thus has the potential to be a useful tool for dietary studies in pre-school children, for example obesity prevention trials. PMID:26784226

  3. Multiple tag labeling method for DNA sequencing

    DOEpatents

    Mathies, Richard A.; Huang, Xiaohua C.; Quesada, Mark A.

    1995-01-01

    A DNA sequencing method described which uses single lane or channel electrophoresis. Sequencing fragments are separated in said lane and detected using a laser-excited, confocal fluorescence scanner. Each set of DNA sequencing fragments is separated in the same lane and then distinguished using a binary coding scheme employing only two different fluorescent labels. Also described is a method of using radio-isotope labels.

  4. Multiple tag labeling method for DNA sequencing

    DOEpatents

    Mathies, R.A.; Huang, X.C.; Quesada, M.A.

    1995-07-25

    A DNA sequencing method is described which uses single lane or channel electrophoresis. Sequencing fragments are separated in the lane and detected using a laser-excited, confocal fluorescence scanner. Each set of DNA sequencing fragments is separated in the same lane and then distinguished using a binary coding scheme employing only two different fluorescent labels. Also described is a method of using radioisotope labels. 5 figs.

  5. High levels of isotope elimination improve precision and allow individual-based measurements of metabolic rates in animals using the doubly labeled water method

    PubMed Central

    Shirai, Masaki; Niizuma, Yasuaki; Yamamoto, Maki; Oda, Emiko; Ebine, Naoyuki; Oka, Nariko; Yoda, Ken

    2015-01-01

    Doubly labeled water (DLW) can be used to measure energy expenditure in free-ranging animals, but questions have been raised about its accuracy in different species or contexts. We investigated whether differences in the extent of isotope elimination affects the precision and accuracy of the DLW method, which can vary according to the experimental design or metabolic rate of the species. Estimated total energy expenditure by the DLW method (TEEdlw) was compared with actual total energy expenditure simultaneously measured via respirometry (TEEresp) in streaked shearwaters Calonectris leucomelas, a pelagic seabird. Subjects were divided into three groups with different experimental conditions: at rest on the ground for 24 h (Group A) or for 48 h (Group B), and at rest on the water for 24 h (Group C). TEEdlw in Group A matched TEEresp, whereas there was an overestimation of TEEdlw in both Groups B and C compared with TEEresp. However, compared with Group A, TEEdlw in Groups B and C had reduced the isotopic analytical variability and thus higher precision. The best regression model (TEEdlw = 1.37 TEEresp − 14.12) showed a high correlation (R2 = 0.82) between TEEdlw and TEEresp and allows a correction factor for field metabolic rates in streaked shearwaters. Our results demonstrate that the commonly made assumption that the DLW method is not appropriate for individual-based estimates may be incorrect in certain circumstances. Although a correction factor may be necessary when using the DLW method to estimate metabolic rate, greater levels of isotope eliminations provides DLW estimates with high precision, which can adequately represent relative individual estimates. Nevertheless, the DLW method, should be used with caution when characterizing interspecies difference of energy expenditures. PMID:26611463

  6. High levels of isotope elimination improve precision and allow individual-based measurements of metabolic rates in animals using the doubly labeled water method.

    PubMed

    Shirai, Masaki; Niizuma, Yasuaki; Yamamoto, Maki; Oda, Emiko; Ebine, Naoyuki; Oka, Nariko; Yoda, Ken

    2015-11-01

    Doubly labeled water (DLW) can be used to measure energy expenditure in free-ranging animals, but questions have been raised about its accuracy in different species or contexts. We investigated whether differences in the extent of isotope elimination affects the precision and accuracy of the DLW method, which can vary according to the experimental design or metabolic rate of the species. Estimated total energy expenditure by the DLW method (TEEdlw) was compared with actual total energy expenditure simultaneously measured via respirometry (TEEresp) in streaked shearwaters Calonectris leucomelas, a pelagic seabird. Subjects were divided into three groups with different experimental conditions: at rest on the ground for 24 h (Group A) or for 48 h (Group B), and at rest on the water for 24 h (Group C). TEEdlw in Group A matched TEEresp, whereas there was an overestimation of TEEdlw in both Groups B and C compared with TEEresp. However, compared with Group A, TEEdlw in Groups B and C had reduced the isotopic analytical variability and thus higher precision. The best regression model (TEEdlw = 1.37 TEEresp - 14.12) showed a high correlation (R(2) = 0.82) between TEEdlw and TEEresp and allows a correction factor for field metabolic rates in streaked shearwaters. Our results demonstrate that the commonly made assumption that the DLW method is not appropriate for individual-based estimates may be incorrect in certain circumstances. Although a correction factor may be necessary when using the DLW method to estimate metabolic rate, greater levels of isotope eliminations provides DLW estimates with high precision, which can adequately represent relative individual estimates. Nevertheless, the DLW method, should be used with caution when characterizing interspecies difference of energy expenditures.

  7. Stable isotope labeling methods for DNA.

    PubMed

    Nelissen, Frank H T; Tessari, Marco; Wijmenga, Sybren S; Heus, Hans A

    2016-08-01

    NMR is a powerful method for studying proteins and nucleic acids in solution. The study of nucleic acids by NMR is far more challenging than for proteins, which is mainly due to the limited number of building blocks and unfavorable spectral properties. For NMR studies of DNA molecules, (site specific) isotope enrichment is required to facilitate specific NMR experiments and applications. Here, we provide a comprehensive review of isotope-labeling strategies for obtaining stable isotope labeled DNA as well as specifically stable isotope labeled building blocks required for enzymatic DNA synthesis. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. PRN 93-3: Labeling Statement Prohibiting Application to Water

    EPA Pesticide Factsheets

    This notice explaining the policy on label statement prohibiting pesticide application to water pertains only to the labeling statement on pesticide products. It does not address the term wetlands as defined with respect to the Clean Water Act.

  9. Accuracy of dietary reference intake predictive equation for estimated energy requirements in female tennis athletes and non-athlete college students: comparison with the doubly labeled water method

    PubMed Central

    Ndahimana, Didace; Lee, Sun-Hee; Kim, Ye-Jin; Son, Hee-Ryoung; Ishikawa-Takata, Kazuko; Park, Jonghoon

    2017-01-01

    BACKGROUND/OBJECTIVES The purpose of this study was to assess the accuracy of a dietary reference intake (DRI) predictive equation for estimated energy requirements (EER) in female college tennis athletes and non-athlete students using doubly labeled water (DLW) as a reference method. MATERIALS/METHODS Fifteen female college students, including eight tennis athletes and seven non-athlete subjects (aged between 19 to 24 years), were involved in the study. Subjects' total energy expenditure (TEE) was measured by the DLW method, and EER were calculated using the DRI predictive equation. The accuracy of this equation was assessed by comparing the EER calculated using the DRI predictive equation (EERDRI) and TEE measured by the DLW method (TEEDLW) based on calculation of percentage difference mean and percentage of accurate prediction. The agreement between the two methods was assessed by the Bland-Altman method. RESULTS The percentage difference mean between the methods was -1.1% in athletes and 1.8% in non-athlete subjects, whereas the percentage of accurate prediction was 37.5% and 85.7%, respectively. In the case of athletic subjects, the DRI predictive equation showed a clear bias negatively proportional to the subjects' TEE. CONCLUSIONS The results from this study suggest that the DRI predictive equation could be used to obtain EER in non-athlete female college students at a group level. However, this equation would be difficult to use in the case of athletes at the group and individual levels. The development of a new and more appropriate equation for the prediction of energy expenditure in athletes is proposed. PMID:28194265

  10. Accuracy of dietary reference intake predictive equation for estimated energy requirements in female tennis athletes and non-athlete college students: comparison with the doubly labeled water method.

    PubMed

    Ndahimana, Didace; Lee, Sun-Hee; Kim, Ye-Jin; Son, Hee-Ryoung; Ishikawa-Takata, Kazuko; Park, Jonghoon; Kim, Eun-Kyung

    2017-02-01

    The purpose of this study was to assess the accuracy of a dietary reference intake (DRI) predictive equation for estimated energy requirements (EER) in female college tennis athletes and non-athlete students using doubly labeled water (DLW) as a reference method. Fifteen female college students, including eight tennis athletes and seven non-athlete subjects (aged between 19 to 24 years), were involved in the study. Subjects' total energy expenditure (TEE) was measured by the DLW method, and EER were calculated using the DRI predictive equation. The accuracy of this equation was assessed by comparing the EER calculated using the DRI predictive equation (EERDRI) and TEE measured by the DLW method (TEEDLW) based on calculation of percentage difference mean and percentage of accurate prediction. The agreement between the two methods was assessed by the Bland-Altman method. The percentage difference mean between the methods was -1.1% in athletes and 1.8% in non-athlete subjects, whereas the percentage of accurate prediction was 37.5% and 85.7%, respectively. In the case of athletic subjects, the DRI predictive equation showed a clear bias negatively proportional to the subjects' TEE. The results from this study suggest that the DRI predictive equation could be used to obtain EER in non-athlete female college students at a group level. However, this equation would be difficult to use in the case of athletes at the group and individual levels. The development of a new and more appropriate equation for the prediction of energy expenditure in athletes is proposed.

  11. Tritium labeling of antisense oligonucleotides by exchange with tritiated water.

    PubMed Central

    Graham, M J; Freier, S M; Crooke, R M; Ecker, D J; Maslova, R N; Lesnik, E A

    1993-01-01

    We describe a simple, efficient, procedure for labeling oligonucleotides to high specific activity (< 1 x 10(8) cpm/mumol) by hydrogen exchange with tritiated water at the C8 positions of purines in the presence of beta-mercaptoethanol, an effective radical scavenger. Approximately 90% of the starting material is recovered as intact, labeled oligonucleotide. The radiolabeled compounds are stable in biological systems; greater than 90% of the specific activity is retained after 72 hr incubation at 37 degrees C in serum-containing media. Data obtained from in vitro cellular uptake experiments using oligonucleotides labeled by this method are similar to those obtained using 35S or 14C-labeled compounds. Because this protocol is solely dependent upon the existence of purine residues, it should be useful for radiolabeling modified as well as unmodified phosphodiester oligonucleotides. Images PMID:8367289

  12. Energy costs of surgery as measured by the doubly labeled water (/sup 2/H/sub 2//sup 18/O) method

    SciTech Connect

    Novick, W.M.; Nusbaum, M.; Stein, T.P.

    1988-01-01

    Energy expenditure before and after surgery was determined in seven patients by the doubly labeled water (/sup 2/H/sub 2//sup 18/O) method (DLW). The values were compared with values obtained by respiratory gas exchange by means of a metabolic measuring cart (MMC). Patients were maintained on total parenteral nutrition before and after trauma. The principal finding was an increase in the rate of CO/sub 2/ production of 11.9 +/- 5.0% after surgery. This corresponds to a 267 +/- increase in energy expenditure (p less than 0.05). No trauma-associated change in energy expenditure was found with the MMC. The correlation of preoperative values from MMC and DLW was not statistically significant (r = 0.25), nor was the correlation of MMC and the Harris-Benedict equation, but the correlation of DLW with Harris-Benedict equation was statistically significant (r = 0.73, p less than 0.05). We suggest that the discrepancy is because the DLW method measures the cumulative energy expenditure over a period, whereas the MMC gives a spot measurement.

  13. Application of the two-sample doubly labelled water method alters behaviour and affects estimates of energy expenditure in black-legged kittiwakes.

    PubMed

    Schultner, Jannik; Welcker, Jorg; Speakman, John R; Nordøy, Erling S; Gabrielsen, Geir W

    2010-09-01

    Despite the widespread use of the doubly labelled water (DLW) method in energetic studies of free-ranging animals, effects of the method on study animals are rarely assessed. We studied behavioural effects of two alternative DLW protocols. During two consecutive breeding seasons, 42 parent black-legged kittiwakes received either the commonly used two-sample (TS) or the less invasive single-sample (SS) DLW treatment. A third group served as a non-treated control. We evaluated the effect of treatment with respect to the time birds took to return to their nest after treatment and recaptures, and the nest attendance during DLW measurement periods. We found that TS kittiwakes took on average 20 times longer to return to their nest than SS kittiwakes after initial treatment, and nest attendance was reduced by about 40% relative to control birds. In contrast, nest attendance did not differ between control and SS kittiwakes. Estimates of energy expenditure of SS kittiwakes exceeded those of TS kittiwakes by 15%. This difference was probably caused by TS birds remaining inactive for extended time periods while at sea. Our results demonstrate that the common assumption that the TS DLW method has little impact on the behaviour of study subjects is in some circumstances fallacious. Estimates of energy expenditure derived by the SS approach may thus more accurately reflect unbiased rates of energy expenditure. However, the choice of protocol may be a trade-off between their impact on behaviour, and hence accuracy, and their differences in precision. Adopting procedures that minimize the impact of TS protocols may be useful.

  14. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... AND REGULATIONS UNDER FUR PRODUCTS LABELING ACT Regulations § 301.27 Label and method of affixing. At all times during the marketing of a fur product the required label shall have a minimum dimension...

  15. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... AND REGULATIONS UNDER FUR PRODUCTS LABELING ACT Regulations § 301.27 Label and method of affixing. At all times during the marketing of a fur product the required label shall have a minimum dimension...

  16. Method of making colloid labeled with radionuclide

    DOEpatents

    Atcher, Robert W.; Hines, John J.

    1991-01-01

    A ferric hydroxide colloid having an alpha-emitting radionuclide essentially on the outer surfaces and a method of forming same. The method includes oxidizing a ferrous hydroxide to ferric hydroxide in the presence of a preselected radionuclide to form a colloid having the radionuclide on the outer surface thereof, and thereafter washing the colloid, and suspending the washed colloid in a suitable solution. The labelled colloid is useful in cancer therapy and for the treatment of inflamed joints.

  17. Colloid labelled with radionuclide and method

    DOEpatents

    Atcher, Robert W.; Hines, John J.

    1990-01-01

    A ferric hydroxide colloid having an alpha-emitting radionuclide essentially on the outer surfaces and a method of forming same. The method includes oxidizing a ferrous hydroxide to ferric hydroxide in the presence of a preselected radionuclide to form a colloid having the radionuclide on the outer surface thereof, and thereafter washing the colloid, and suspending the washed colloid in a suitable solution. The labelled colloid is useful in cancer therapy and for the treatment of inflamed joints.

  18. Colloid labelled with radionuclide and method

    DOEpatents

    Atcher, R.W.; Hines, J.J.

    1990-11-13

    A ferric hydroxide colloid having an alpha-emitting radionuclide essentially on the outer surfaces and a method of forming same. The method includes oxidizing a ferrous hydroxide to ferric hydroxide in the presence of a preselected radionuclide to form a colloid having the radionuclide on the outer surface thereof, and thereafter washing the colloid, and suspending the washed colloid in a suitable solution. The labelled colloid is useful in cancer therapy and for the treatment of inflamed joints. No Drawings

  19. Walking as a Contributor to Physical Activity in Healthy Older Adults: 2 Week Longitudinal Study Using Accelerometry and the Doubly Labeled Water Method

    PubMed Central

    Bonomi, Alberto G; Westerterp, Klaas R

    2016-01-01

    Background Physical activity is recommended to promote healthy aging. Defining the importance of activities such as walking in achieving higher levels of physical activity might provide indications for interventions. Objective To describe the importance of walking in achieving higher levels of physical activity in older adults. Methods The study included 42 healthy subjects aged between 51 and 84 years (mean body mass index 25.6 kg/m2 [SD 2.6]). Physical activity, walking, and nonwalking activity were monitored with an accelerometer for 2 weeks. Physical activity was quantified by accelerometer-derived activity counts. An algorithm based on template matching and signal power was developed to classify activity counts into nonwalking counts, short walk counts, and long walk counts. Additionally, in a subgroup of 31 subjects energy expenditure was measured using doubly labeled water to derive physical activity level (PAL). Results Subjects had a mean PAL of 1.84 (SD 0.19, range 1.43-2.36). About 20% of the activity time (21% [SD 8]) was spent walking, which accounted for about 40% of the total counts (43% [SD 11]). Short bouts composed 83% (SD 9) of walking time, providing 81% (SD 11) of walking counts. A stepwise regression model to predict PAL included nonwalking counts and short walk counts, explaining 58% of the variance of PAL (standard error of the estimate=0.12). Walking activities produced more counts per minute than nonwalking activities (P<.001). Long walks produced more counts per minute than short walks (P=.001). Nonwalking counts were independent of walking counts (r=−.05, P=.38). Conclusions Walking activities are a major contributor to physical activity in older adults. Walking activities occur at higher intensities than nonwalking activities, which might prevent individuals from engaging in more walking activity. Finally, subjects who engage in more walking activities do not tend to compensate by limiting nonwalking activities. Trial Registration

  20. WaterSense Labeled Homes Quick Reference Guide

    EPA Pesticide Factsheets

    Green building has grown from a niche market to a savvy business strategy. WaterSense labeled homes capitalize on consumer demand by offering homeowners a whole-house solution to help them save water, energy, and money.

  1. Kinugasa reactions in water: from green chemistry to bioorthogonal labelling.

    PubMed

    Chigrinova, Mariya; MacKenzie, Douglas A; Sherratt, Allison R; Cheung, Lawrence L W; Pezacki, John Paul; Pezacki, Paul

    2015-04-16

    The Kinugasa reaction has become an efficient method for the direct synthesis of β-lactams from substituted nitrones and copper(I) acetylides. In recent years, the reaction scope has been expanded to include the use of water as the solvent, and with micelle-promoted [3+2] cycloadditions followed by rearrangement furnishing high yields of β-lactams. The high yields of stable products under aqueous conditions render the modified Kinugasa reaction amenable to metabolic labelling and bioorthogonal applications. Herein, the development of methods for use of the Kinugasa reaction in aqueous media is reviewed, with emphasis on its potential use as a bioorthogonal coupling strategy.

  2. 16 CFR 300.11 - Improper methods of labeling.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) Crowding, intermingling, or obscuring with designs, vignettes, or other written, printed or graphic matter. ... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Improper methods of labeling. 300.11 Section... AND REGULATIONS UNDER THE WOOL PRODUCTS LABELING ACT OF 1939 Labeling § 300.11 Improper methods...

  3. 16 CFR 300.11 - Improper methods of labeling.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Improper methods of labeling. 300.11 Section 300.11 Commercial Practices FEDERAL TRADE COMMISSION REGULATIONS UNDER SPECIFIC ACTS OF CONGRESS RULES AND REGULATIONS UNDER THE WOOL PRODUCTS LABELING ACT OF 1939 Labeling § 300.11 Improper methods...

  4. Refining cotton-wick method for 15N plant labelling.

    NASA Astrophysics Data System (ADS)

    Fustec, Joëlle; Mahieu, Stéphanie

    2010-05-01

    The symbiosis Fabaceae/Rhizobiaceae plays a critical role in the nitrogen cycle. It gives the plant the ability to fix high amounts of atmospheric N. A part of this N can be transferred to the soil via rhizodeposition. The contribution of Fabaceae to the soil N pool is difficult to measure, since it is necessary for assessing N benefits for other crops, for soil biological activity, and for reducing water pollution in sustainable agriculture (Fustec, 2009). The aim of this study was to test and improve the reliability of the 15N cotton-wick method for measuring the soil N derived from plant rhizodeposition (Mahieu et al., 2007). The effects of the concentration of the 15N-urea labelling solution and of the feeding frequency (continuous or pulses) on the assessment of nitrogen rhizodeposition were studied in two greenhouse experiments using the field pea (Pisum sativum L.) and the non-nodulating isoline P2. The plant parts and the soil were prepared for 15N:14N measurements for assessing N rhizodeposition (Mahieu et al., 2009). The fraction of plants' belowground nitrogen allocated to rhizodeposition in both Frisson pea and P2 was 20 to more than 50% higher when plants were labelled continuously than when they were labelled using fortnightly pulses. Our results suggested that when 15N root enrichment was high, nitrogen rhizodeposition was underestimated only for plants that were 15N-fed by fortnightly pulses, and not in plants 15N-fed continuously. This phenomenon was especially observed for plants relying on symbiotic N fixation for N acquisition; it may be linked to the concentration of the labelling solution. In conclusion, N rhizodeposition assessment was strongly influenced by the 15N-feeding frequency and the concentration of the labelling solution. The estimation of N rhizodeposition was more reliable when plants were labelled continuously with a dilute solution of 15N urea. Fustec et al. 2009. Agron. Sustain. Dev., DOI 10.1051/agro/2009003, in press. Mahieu

  5. [Progress in stable isotope labeled quantitative proteomics methods].

    PubMed

    Zhou, Yuan; Shan, Yichu; Zhang, Lihua; Zhang, Yukui

    2013-06-01

    Quantitative proteomics is an important research field in post-genomics era. There are two strategies for proteome quantification: label-free methods and stable isotope labeling methods which have become the most important strategy for quantitative proteomics at present. In the past few years, a number of quantitative methods have been developed, which support the fast development in biology research. In this work, we discuss the progress in the stable isotope labeling methods for quantitative proteomics including relative and absolute quantitative proteomics, and then give our opinions on the outlook of proteome quantification methods.

  6. Comparison of doubly labeled water with respirometry at low- and high-activity levels

    SciTech Connect

    Westerterp, K.R.; Brouns, F.; Saris, W.H.; ten Hoor, F.

    1988-07-01

    In previous studies the doubly labeled water method for measuring energy expenditure in free-living humans has been validated against respirometry under sedentary conditions. In the present investigation, energy expenditure is measured simultaneously with doubly labeled water and respirometry at low- and high-activity levels. Over 6 days, five subjects were measured doing mainly sedentary activities like desk work; their average daily metabolic rate was 1.40 +/- 0.09 (SD) times sleeping metabolic rate. Four subjects were measured twice over 3.5 days, including 2 days with heavy bicycle ergometer work, resulting in an average daily metabolic rate of 2.61 +/- 0.25 (SD) times sleeping metabolic rate. At the low-activity level, energy expenditures from the doubly labeled water method were on the average 1.4 +/- 3.9% (SD) larger than those from respirometry. At the high-activity level, the doubly labeled water method yielded values that were 1.0 +/- 7.0% (SD) lower than those from respirometry. Results demonstrate the utility of the doubly labeled water method for the determination of energy expenditure in the range of activity levels in daily life.

  7. The Assessment of Total Energy Expenditure During a 14-Day In-Season Period of Professional Rugby League Players Using the Doubly Labelled Water Method.

    PubMed

    Morehen, James Cameron; Bradley, Warren Jeremy; Clarke, Jon; Twist, Craig; Hambly, Catherine; Speakman, John Roger; Morton, James Peter; Close, Graeme Leonard

    2016-10-01

    Rugby League is a high-intensity collision sport competed over 80 min. Training loads are monitored to maximize recovery and assist in the design of nutritional strategies although no data are available on the total energy expenditure (TEE) of players. We therefore assessed resting metabolic rate (RMR) and TEE in six Super League players over 2 consecutive weeks in-season including one game per week. Fasted RMR was assessed followed by a baseline urine sample before oral administration of a bolus dose of hydrogen (deuterium (2)H) and oxygen ((18)O) stable isotopes in the form of water ((2)H2(18)O). Every 24 hr thereafter, players provided urine for analysis of TEE via DLW method. Individual training load was quantified using session rating of perceived exertion (sRPE) and data were analyzed using magnitude-based inferences. There were unclear differences in RMR between forwards and backs (7.7 ± 0.5 cf. 8.0 ± 0.3 MJ, respectively). Indirect calorimetry produced RMR values most likely lower than predictive equations (7.9 ± 0.4 cf. 9.2 ± 0.4 MJ, respectively). A most likely increase in TEE from Week 1 to 2 was observed (17.9 ± 2.1 cf. 24.2 ± 3.4 MJ) explained by a most likelyincrease in weekly sRPE (432 ± 19 cf. 555 ± 22 AU), respectively. The difference in TEE between forward and backs was unclear (21.6 ± 4.2 cf. 20.5 ± 4.9 MJ, respectively). We report greater TEE than previously reported in rugby that could be explained by the ability of DLW to account for all match and training-related activities that contributes to TEE.

  8. Water treatment method

    DOEpatents

    Martin, F.S.; Silver, G.L.

    1991-04-30

    A method is described for reducing the concentration of any undesirable metals dissolved in contaminated water, such as waste water. The method involves uniformly reacting the contaminated water with an excess amount of solid particulate calcium sulfite to insolubilize the undesirable metal ions, followed by removal thereof and of the unreacted calcium sulfite.

  9. Water treatment method

    DOEpatents

    Martin, Frank S.; Silver, Gary L.

    1991-04-30

    A method for reducing the concentration of any undesirable metals dissolved in contaminated water, such as waste water. The method involves uniformly reacting the contaminated water with an excess amount of solid particulate calcium sulfite to insolubilize the undesirable metal ions, followed by removal thereof and of the unreacted calcium sulfite.

  10. Water treatment method

    SciTech Connect

    Martin, F.S.; Silver, G.L.

    1990-02-02

    A method for reducing the concentration of many undesirable metals dissolved in contaminated water, such as waste water. The method involves uniformly reacting the contaminated water with an excess amount of solid particulate calcium sulfite to insolubilize the undesirable metal ions, followed by removal thereof and of the unreacted calcium sulfite. 1 tab.

  11. Water-budget methods

    USGS Publications Warehouse

    Healy, Richard W.; Scanlon, Bridget R.

    2010-01-01

    A water budget is an accounting of water movement into and out of, and storage change within, some control volume. Universal and adaptable are adjectives that reflect key features of water-budget methods for estimating recharge. The universal concept of mass conservation of water implies that water-budget methods are applicable over any space and time scales (Healy et al., 2007). The water budget of a soil column in a laboratory can be studied at scales of millimeters and seconds. A water-budget equation is also an integral component of atmospheric general circulation models used to predict global climates over periods of decades or more. Water-budget equations can be easily customized by adding or removing terms to accurately portray the peculiarities of any hydrologic system. The equations are generally not bound by assumptions on mechanisms by which water moves into, through, and out of the control volume of interest. So water-budget methods can be used to estimate both diffuse and focused recharge, and recharge estimates are unaffected by phenomena such as preferential flow paths within the unsaturated zone.Water-budget methods represent the largest class of techniques for estimating recharge. Most hydrologic models are derived from a water-budget equation and can therefore be classified as water-budget models. It is not feasible to address all water-budget methods in a single chapter. This chapter is limited to discussion of the “residual” water-budget approach, whereby all variables in a water-budget equation, except for recharge, are independently measured or estimated and recharge is set equal to the residual. This chapter is closely linked with Chapter 3, on modeling methods, because the equations presented here form the basis of many models and because models are often used to estimate individual components in water-budget studies. Water budgets for streams and other surface-water bodies are addressed in Chapter 4. The use of soil-water budgets and

  12. Estimating plant water uptake source depths with optimized stable water isotope labeling

    NASA Astrophysics Data System (ADS)

    Seeger, Stefan; Weiler, Markus

    2016-04-01

    Depth profiles of pore water stable isotopes in soils in conjunction with measurements of stable water isotopes (SWI) in plant transpiration allow the estimation of the contributions of different soil depths to plant water uptake (PWU).
 However, SWI depth profiles that result from the variations of SWI in natural precipitation may lead to highly ambiguous results, i.e. the same SWI signature in transpiration could result from different PWU patterns or SWI depth profiles. The aim of this study was to find an optimal stable water isotope depth profile to estimate plant water uptake patterns and to compare different PWU source depth estimation methods. We used a new soil water transport model including fractionation effects of SWI and exchange between the vapor and liquid phase to simulate different irrigation scenarios. Different amounts of water with differing SWI signatures (glacier melt water, summer precipitation water, deuterated water) were applied in order to obtain a wide variety of SWI depth profiles. Based on these simulated SWI depth profiles and a set of hypothetical PWU patterns, the theoretical SWI signatures of the respective plant transpiration were computed. In the next step, two methods - Bayesian isotope mixing models (BIMs) and optimization of a parametric distribution function (beta function) - were used to estimate the PWU patterns from the different SWI depth profiles and their respective SWI signatures in the resulting transpiration. Eventually, the estimated and computed profiles were compared to find the best SWI depth profile and the best method. The results showed, that compared to naturally occurring SWI depth profiles, the application of multiple, in terms of SWI, distinct labeling pulses greatly improves the possible spatial resolution and at the same time reduces the uncertainty of PWU estimates.
 For the PWU patterns which were assumed for this study, PWU pattern estimates based on an optimized parametric distribution function

  13. Doubly labelled water assessment of energy expenditure: principle, practice, and promise.

    PubMed

    Westerterp, Klaas R

    2017-07-01

    The doubly labelled water method for the assessment of energy expenditure was first published in 1955, application in humans started in 1982, and it has become the gold standard for human energy requirement under daily living conditions. The method involves enriching the body water of a subject with heavy hydrogen ((2)H) and heavy oxygen ((18)O), and then determining the difference in washout kinetics between both isotopes, being a function of carbon dioxide production. In practice, subjects get a measured amount of doubly labelled water ((2)H 2(18) O) to increase background enrichment of body water for (18)O of 2000 ppm with at least 180 ppm and background enrichment of body water for (2)H of 150 ppm with 120 ppm. Subsequently, the difference between the apparent turnover rates of the hydrogen and oxygen of body water is assessed from blood-, saliva-, or urine samples, collected at the start and end of the observation interval of 1-3 weeks. Samples are analyzed for (18)O and (2)H with isotope ratio mass spectrometry. The doubly labelled water method is the indicated method to measure energy expenditure in any environment, especially with regard to activity energy expenditure, without interference with the behavior of the subjects. Applications include the assessment of energy requirement from total energy expenditure, validation of dietary assessment methods and validation of physical activity assessment methods with doubly labelled water measured energy expenditure as reference, and studies on body mass regulation with energy expenditure as a determinant of energy balance.

  14. Inter-labeler and intra-labeler variability of condition severity classification models using active and passive learning methods.

    PubMed

    Nissim, Nir; Shahar, Yuval; Elovici, Yuval; Hripcsak, George; Moskovitch, Robert

    2017-04-26

    Labeling instances by domain experts for classification is often time consuming and expensive. To reduce such labeling efforts, we had proposed the application of active learning (AL) methods, introduced our CAESAR-ALE framework for classifying the severity of clinical conditions, and shown its significant reduction of labeling efforts. The use of any of three AL methods (one well known [SVM-Margin], and two that we introduced [Exploitation and Combination_XA]) significantly reduced (by 48% to 64%) condition labeling efforts, compared to standard passive (random instance-selection) SVM learning. Furthermore, our new AL methods achieved maximal accuracy using 12% fewer labeled cases than the SVM-Margin AL method. However, because labelers have varying levels of expertise, a major issue associated with learning methods, and AL methods in particular, is how to best to use the labeling provided by a committee of labelers. First, we wanted to know, based on the labelers' learning curves, whether using AL methods (versus standard passive learning methods) has an effect on the Intra-labeler variability (within the learning curve of each labeler) and inter-labeler variability (among the learning curves of different labelers). Then, we wanted to examine the effect of learning (either passively or actively) from the labels created by the majority consensus of a group of labelers. We used our CAESAR-ALE framework for classifying the severity of clinical conditions, the three AL methods and the passive learning method, as mentioned above, to induce the classifications models. We used a dataset of 516 clinical conditions and their severity labeling, represented by features aggregated from the medical records of 1.9 million patients treated at Columbia University Medical Center. We analyzed the variance of the classification performance within (intra-labeler), and especially among (inter-labeler) the classification models that were induced by using the labels provided by seven

  15. Comparative examination of probe labeling methods for microarray hybridization

    NASA Astrophysics Data System (ADS)

    Burke, David I.; Woodward, Karen; Setterquist, Robert A.; Kawasaki, Ernest S.

    2001-06-01

    For detection of differential gene expression, confocal laser based scanners are now capable of analyzing microarrays using one to five wavelengths. This allows investigators to choose among several labeling methods. Here we compare direct incorporation and indirect methods (amino-allyl and dendrimers) for labeling cDNA probes. We assessed reproducible sensitivity of each probe preparation method in two ways. First, by comparing hybridization intensities for limit of signal detection and second by measuring the lowest detectable concentration of a known ratio of mixed DNA (spikes). Limit of detection assay was done using arrays of mixed targets consisting of a serially diluted human specific gene fragment (HU1) and an undiluted DNA of chloramphenicol acetyl tranferase (CAT) gene. Then, individual single target arrays of CAT and HU1 DNA were used to determine the lowest detectable spike ratio of each labeling method. The results of this study will be presented and their significance for the analysis of microarrays will be discussed.

  16. Novel method and applications for labeling and identifying lymph nodes

    NASA Astrophysics Data System (ADS)

    Kiraly, Atilla P.; Naidich, David P.; Guendel, Lutz; Zhang, Li; Novak, Carol L.

    2007-03-01

    The lymphatic system comprises a series of interconnected lymph nodes that are commonly distributed along branching or linearly oriented anatomic structures. Physicians must evaluate lymph nodes when staging cancer and planning optimal paths for nodal biopsy. This process requires accurately determining the lymph node's position with respect to major anatomical landmarks. In an effort to standardize lung cancer staging, The American Joint Committee on Cancer (AJCC) has classified lymph nodes within the chest into 4 groups and 14 sub groups. We present a method for automatically labeling lymph nodes according to this classification scheme, in order to improve the speed and accuracy of staging and biopsy planning. Lymph nodes within the chest are clustered around the major blood vessels and the airways. Our fully automatic labeling method determines the nodal group and sub-group in chest CT data by use of computed airway and aorta centerlines to produce features relative to a given node location. A classifier then determines the label based upon these features. We evaluate the efficacy of the method on 10 chest CT datasets containing 86 labeled lymph nodes. The results are promising with 100% of the nodes assigned to the correct group and 76% to the correct sub-group. We anticipate that additional features and training data will further improve the results. In addition to labeling, other applications include automated lymph node localization and visualization. Although we focus on chest CT data, the method can be generalized to other regions of the body as well as to different imaging modalities.

  17. Validation of doubly labeled water for measuring energy expenditure during parenteral nutrition

    SciTech Connect

    Schoeller, D.A.; Kushner, R.F.; Jones, P.J.

    1986-08-01

    The doubly labeled water method was compared with intake-balance for measuring energy expenditure in five patients receiving total parenteral nutrition (TPN). Because parenteral solutions were isotopically different from local water, patients had to be placed on TPN at least 10 days before the metabolic period. Approximately 0.1 g 2H2O and 0.25 g H2(18)O per kg total body water were given orally. We collected saliva before, 3 h, and 4 h after the dose for measurement of total body water and urine before, 1 day, and 14 days after the dose for measurement of isotope eliminations. On day 14, total body weight was remeasured and change in body energy stores was calculated, assuming constant hydration. Intake was assessed from weights of TPN fluids plus dietary record for any oral intake. Energy expenditure from doubly labeled water (+/- SD) averaged 3 +/- 6% greater than intake-balance. Doubly labeled water method is a noninvasive, nonrestrictive method for measuring energy expenditure in patients receiving TPN.

  18. Method for labelling leucocytes with indium In-111 oxine

    SciTech Connect

    Kaminsky, D.

    1992-03-03

    This patent describes an improved method for radio-labelling leucocytes with Indium In-111 oxine. It comprises separating the leucocytes from whole blood for obtaining separated leucocytes mixed with residual red blood cells; and then labelling the separated leucocytes with Indium In-111 oxine; wherein the improvement comprises the following further step: depleting residual red blood cells from the separated leucocytes by resuspending the leucocytes in an isotonic saline solution, then rocking the resuspended leucocytes for causing the leucocytes to preferentially settle out, and then removing residual red blood cells which remain suspended within the supernatant isotonic saline solution.

  19. New method of iodine labelling of peptide hormones

    SciTech Connect

    Escher, E.

    1984-01-01

    Usually peptide hormones and related compounds are radioactively labelled with iodine on tyrosine residues of the peptide. However many peptide hormones do not contain tyrosine or the iodinated tyrosine interferes with the biological properties. In order to circumvent these and other problems, a general method is proposed which allows the introduction of iodine into the para-position of phenylalanine with a modified Sandmeyer procedure. This last-step modification together with HPLC purification permits the obtention of carrier-free and metabolically stable labelled products with maximal specific activity possible. The model has been carried out on several peptide-models like angiotensin II, endorphine and head activator peptide.

  20. Volume-labeled nanoparticles and methods of preparation

    DOEpatents

    Wang, Wei; Gu, Baohua; Retterer, Scott T; Doktycz, Mitchel J

    2015-04-21

    Compositions comprising nanosized objects (i.e., nanoparticles) in which at least one observable marker, such as a radioisotope or fluorophore, is incorporated within the nanosized object. The nanosized objects include, for example, metal or semi-metal oxide (e.g., silica), quantum dot, noble metal, magnetic metal oxide, organic polymer, metal salt, and core-shell nanoparticles, wherein the label is incorporated within the nanoparticle or selectively in a metal oxide shell of a core-shell nanoparticle. Methods of preparing the volume-labeled nanoparticles are also described.

  1. Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture

    DOEpatents

    Sanfilippo, Antonio; Calapristi, Augustin J.; Crow, Vernon L.; Hetzler, Elizabeth G.; Turner, Alan E.

    2009-12-22

    Document clustering methods, document cluster label disambiguation methods, document clustering apparatuses, and articles of manufacture are described. In one aspect, a document clustering method includes providing a document set comprising a plurality of documents, providing a cluster comprising a subset of the documents of the document set, using a plurality of terms of the documents, providing a cluster label indicative of subject matter content of the documents of the cluster, wherein the cluster label comprises a plurality of word senses, and selecting one of the word senses of the cluster label.

  2. Accuracy of a combined heart rate and motion sensor for assessing energy expenditure in free-living adults during a double-blind crossover caffeine trial using doubly labeled water as the reference method.

    PubMed

    Silva, A M; Santos, D A; Matias, C N; Júdice, P B; Magalhães, J P; Ekelund, U; Sardinha, L B

    2015-01-01

    A combined heart rate (HR) and motion sensor (Actiheart) has been proposed as an accurate method for assessing total energy expenditure (TEE) and physical activity energy expenditure (PAEE). However, the extent to which factors such as caffeine may affect the accuracy by which the estimated HR-related PAEE contribution will affect TEE and PAEE estimates is unknown. Therefore, we examined the validity of Actiheart in estimating TEE and PAEE in free-living adults under a caffeine trial compared with doubly labeled water (DLW) as reference criterion. Using a double-blind crossover trial (Clinicaltrials.gov ID: #NCT01477294) with two conditions (4-day each with a 3-day-washout period), randomly ordered as caffeine (5 mg/kg per day) and placebo (malt-dextrine) intake, TEE was measured by DLW in 17 physically active men (20-38 years) who were non-caffeine users. In each condition, resting energy expenditure (REE) was assessed by indirect calorimetry and PAEE was calculated as (TEE-(REE+0.1 TEE)). Simultaneously, PAEE and TEE were estimated by Actiheart using an individual calibration (ACC+HRstep). Under caffeine, ACC+HRstep explained 76 and 64% of TEE and PAEE from DLW, respectively; corresponding results for the placebo condition were 82 and 66%. No mean bias was found between ACC+HRstep and DLW for TEE (caffeine:-468 kJ per day; placebo:-407 kJ per day), although PAEE was slightly underestimated (caffeine:-856 kJ per day; placebo:-1147 kJ per day). Similar limits of agreement were observed in both conditions ranging from -2066 to 3002 and from -3488 to 1776 kJ per day for TEE and PAEE, respectively. Regardless of caffeine intake, the combined HR and motion sensor is valid for estimating free-living energy expenditure in a group of healthy men but is less accurate for an individual assessment.

  3. New Methods for Labeling RGD Peptides with Bromine-76

    PubMed Central

    Lang, Lixin; Li, Weihua; Jia, Hong-Mei; Fang, De-Cai; Zhang, Shushu; Sun, Xilin; Zhu, Lei; Ma, Ying; Shen, Baozhong; Kiesewetter, Dale O.; Niu, Gang; Chen, Xiaoyuan

    2011-01-01

    Direct bromination of the tyrosine residues of peptides and antibodies with bromine-76, to create probes for PET imaging, has been reported. For peptides that do not contain tyrosine residues, however, a prosthetic group is required to achieve labeling via conjugation to other functional groups such as terminal α-amines or lysine ε-amines. The goal of this study was to develop new strategies for labeling small peptides with Br-76 using either a direct labeling method or a prosthetic group, depending on the available functional group on the peptides. A new labeling agent, N-succinimidyl-3-[76Br]bromo-2,6-dimethoxybenzoate ([76Br]SBDMB) was prepared for cyclic RGD peptide labeling. N-succinimidyl-2, 6-dimethoxybenzoate was also used to pre-attach a 2, 6-dimethoxybenzoyl (DMB) moiety to the peptide, which could then be labeled with Br-76. A competitive cell binding assay was performed to determine the binding affinity of the brominated peptides. PET imaging of U87MG human glioblastoma xenografted mice was performed using [76Br]-BrE[c(RGDyK)]2 and [76Br]-BrDMB-E[c(RGDyK)]2. An ex vivo biodistribution assay was performed to confirm PET quantification. The mechanisms of bromination reaction between DMB-c(RGDyK) and the brominating agent CH3COOBr were investigated with the SCRF-B3LYP/6-31G* method with the Gaussian 09 program package. The yield for direct labeling of c(RGDyK) and E[c(RGDyK)]2 using chloramine-T and peracetic acid at ambient temperature was greater than 50%. The yield for [76Br]SBDMB was over 60% using peracetic acid. The conjugation yields for labeling c(RGDfK) and c(RGDyK) were over 70% using the prosthetic group at room temperature. Labeling yield for pre-conjugated peptides was over 60%. SDMB conjugation and bromination did not affect the binding affinity of the peptides with integrin receptors. Both [76Br]Br-E[c(RGDyK)]2 and [76Br]BrDMB-E[c(RGDyK)]2 showed high tumor uptake in U87MG tumor bearing mice. The specificity of the imaging tracers was

  4. Labeled ALPHA4BETA2 ligands and methods therefor

    DOEpatents

    Mukherjee, Jogeshwar; Pichika, Ramaiah; Potkin, Steven; Leslie, Frances; Chattopadhyay, Sankha

    2013-02-19

    Contemplated compositions and methods are employed to bind in vitro and in vivo to an .alpha.4.beta.2 nicotinic acetylcholine receptor in a highly selective manner. Where such compounds are labeled, compositions and methods employing such compounds can be used for PET and SPECT analysis. Alternatively, and/or additionally contemplated compounds can be used as antagonists, partial agonists or agonists in the treatment of diseases or conditions associated with .alpha.4.beta..beta.2 dysfunction.

  5. Pulsed EPR Determination of Water Accessibility to Spin-Labeled Amino Acid Residues in LHCIIb

    PubMed Central

    Volkov, A.; Dockter, C.; Bund, T.; Paulsen, H.; Jeschke, G.

    2009-01-01

    Membrane proteins reside in a structured environment in which some of their residues are accessible to water, some are in contact with alkyl chains of lipid molecules, and some are buried in the protein. Water accessibility of residues may change during folding or function-related structural dynamics. Several techniques based on the combination of pulsed electron paramagnetic resonance (EPR) with site-directed spin labeling can be used to quantify such water accessibility. Accessibility parameters for different residues in major plant light-harvesting complex IIb are determined by electron spin echo envelope modulation spectroscopy in the presence of deuterated water, deuterium contrast in transversal relaxation rates, analysis of longitudinal relaxation rates, and line shape analysis of electron-spin-echo-detected EPR spectra as well as by the conventional techniques of measuring the maximum hyperfine splitting and progressive saturation in continuous-wave EPR. Systematic comparison of these parameters allows for a more detailed characterization of the environment of the spin-labeled residues. These techniques are applicable independently of protein size and require ∼10–20 nmol of singly spin-labeled protein per sample. For a residue close to the N-terminus, in a domain unresolved in the existing x-ray structures of light-harvesting complex IIb, all methods indicate high water accessibility. PMID:19186148

  6. Exercise, energy expenditure and energy balance, as measured with doubly labelled water.

    PubMed

    Westerterp, Klaas R

    2017-07-20

    The doubly labelled water method for the measurement of total daily energy expenditure (TDEE) over 1-3 weeks under daily living conditions is the indicated method to study effects of exercise and extreme environments on energy balance. Subjects consume a measured amount of doubly labelled water (2H2 18O) to increase background enrichment of body water for 18O and 2H, and the subsequent difference in elimination rate between 18O and 2H, as measured in urine, saliva or blood samples, is a measure for carbon dioxide production and thus allows calculation of TDEE. The present review describes research showing that physical activity level (PAL), calculated as TDEE (assessed with doubly labelled water) divided by resting energy expenditure (REE, PAL = TDEE/REE), reaches a maximum value of 2·00-2·40 in subjects with a vigorously active lifestyle. Higher PAL values, while maintaining energy balance, are observed in professional athletes consuming additional energy dense foods to compete at top level. Exercise training can increase TDEE/REE in young adults to a value of 2·00-2·40, when energy intake is unrestricted. Furthermore, the review shows an exercise induced increase in activity energy expenditure can be compensated by a reduction in REE and by a reduction in non-exercise physical activity, especially at a negative energy balance. Additionally, in untrained subjects, an exercise-induced increase in activity energy expenditure is compensated by a training-induced increase in exercise efficiency.

  7. SIMPLE: a sequential immunoperoxidase labeling and erasing method.

    PubMed

    Glass, George; Papin, Jason A; Mandell, James W

    2009-10-01

    The ability to simultaneously visualize expression of multiple antigens in cells and tissues can provide powerful insights into cellular and organismal biology. However, standard methods are limited to the use of just two or three simultaneous probes and have not been widely adopted for routine use in paraffin-embedded tissue. We have developed a novel approach called sequential immunoperoxidase labeling and erasing (SIMPLE) that enables the simultaneous visualization of at least five markers within a single tissue section. Utilizing the alcohol-soluble peroxidase substrate 3-amino-9-ethylcarbazole, combined with a rapid non-destructive method for antibody-antigen dissociation, we demonstrate the ability to erase the results of a single immunohistochemical stain while preserving tissue antigenicity for repeated rounds of labeling. SIMPLE is greatly facilitated by the use of a whole-slide scanner, which can capture the results of each sequential stain without any information loss.

  8. Use of doubly labeled water technique in soldiers training for jungle warfare.

    PubMed

    Forbes-Ewan, C H; Morrissey, B L; Gregg, G C; Waters, D R

    1989-07-01

    The doubly labeled water method was used to estimate the energy expended by four members of an Australian Army platoon (34 soldiers) engaged in training for jungle warfare. Each subject received an oral isotope dose sufficient to raise isotope levels by 200-250 (18O) and 100-120 ppm (2H). The experimental period was 7 days. Concurrently, a factorial estimate of the energy expenditure of the platoon was conducted. Also, a food intake-energy balance study was conducted for the platoon. Mean daily energy expenditure by the doubly labeled water method was 4,750 kcal (range 4,152-5,394 kcal). The factorial estimate of mean daily energy expenditure was 4,535 kcal. Because of inherent inaccuracies in the food intake-energy balance technique, we were able to conclude only that energy expenditure, as measured by this method, was greater than the estimated mean daily intake of 4,040 kcal. The doubly labeled water technique was well tolerated, is noninvasive, and appears to be suitable in a wide range of field applications.

  9. Use of doubly labeled water technique in soldiers training for jungle warfare

    SciTech Connect

    Forbes-Ewan, C.H.; Morrissey, B.L.; Gregg, G.C.; Waters, D.R. )

    1989-07-01

    The doubly labeled water method was used to estimate the energy expended by four members of an Australian Army platoon (34 soldiers) engaged in training for jungle warfare. Each subject received an oral isotope dose sufficient to raise isotope levels by 200-250 ({sup 18}O) and 100-120 ppm ({sup 2}H). The experimental period was 7 days. Concurrently, a factorial estimate of the energy expenditure of the platoon was conducted. Also, a food intake-energy balance study was conducted for the platoon. Mean daily energy expenditure by the doubly labeled water method was 4,750 kcal (range 4,152-5,394 kcal). The factorial estimate of mean daily energy expenditure was 4,535 kcal. Because of inherent inaccuracies in the food intake-energy balance technique, we were able to conclude only that energy expenditure, as measured by this method, was greater than the estimated mean daily intake of 4,040 kcal. The doubly labeled water technique was well tolerated, is noninvasive, and appears to be suitable in a wide range of field applications.

  10. Simplified method to substantiate SPF labeling for sunscreen products.

    PubMed

    Sayre, Robert M; Stanfield, Joseph; Lott, Dennis L; Dowdy, John C

    2003-10-01

    Worldwide, sunscreen sun protection factor (SPF) testing is based on 30-year-old technology. At the time the SPF test came into being, the highest SPFs available were in the 6-8 range. The SPF test is reasonably accurate for SPFs up to 15, but is much less reliable for measuring SPFs of 30 and higher. The method we propose addresses two primary reasons for this unreliability: 1) difficulties in applying products uniformly and 2) the subjectivity and variability of perception in evaluating and grading responses to UV doses. Our proposed SPF substantiation method differs from the current SPF test in that sunscreen-protected test sites receive the same UV dose in four uniformly spaced sub-sites, which are graded as passing if no response is seen or failing if any response is seen. The response may be tanning, erythema, or a combination of both. To demonstrate the method, two commercial products with labeled SPFs of 30 and 45 and the P2 sunscreen standard were tested at two different laboratories. The SPF 30 product and SPF 15 standard were shown to be correctly labeled. However, it is questionable as to whether the SPF 45 product provides protection against 45 minimal erythema doses. Our proposed SPF substantiation method is not dependent on subjective evaluation of responses, accounts for non-uniform product application, and provides a conservative estimate of sunscreen protection. The method consists of a systematic repetition of identical tests that are considerably more rigorous than the current methods that are based on single data points per test subject. While the current SPF test remains necessary and valuable as a dose ranging tool, we propose that this SPF substantiation method supersede the old method for final SPF label determination.

  11. A simple method for labeling CT images with respiratory states

    SciTech Connect

    Berlinger, Kajetan; Sauer, Otto; Vences, Lucia; Roth, Michael

    2006-09-15

    A method is described for labeling CT images with their respiratory state by a needle, connected to the patient's chest/abdomen. By means of a leverage the needle follows the abdominal respiratory motion. The needle is visible as a blurred spot in every CT slice. The method was tested with nine patients. A series of volume scans during free breathing was performed. The detected positions of the moving needle in every single slice were compared to each other thus enabling respiratory state assignment. The tool is an inexpensive alternative to complex respiratory measuring tools for four dimensional (4D) CT and was greatly accepted in the clinic due to its simplicity.

  12. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Required label and method of affixing. 300.5 Section 300.5 Commercial Practices FEDERAL TRADE COMMISSION REGULATIONS UNDER SPECIFIC ACTS OF CONGRESS RULES AND REGULATIONS UNDER THE WOOL PRODUCTS LABELING ACT OF 1939 Labeling § 300.5 Required label...

  13. Inspection and Verification Guidance for WaterSense Labeled New Homes (Version 1.2)

    EPA Pesticide Factsheets

    A new home must be built by a WaterSense builder partner, certified by an EPA licensed certification provider, and meet all of the applicable criteria in the specification to become a WaterSense labeled new home.

  14. WaterSense Labeled Weather-Based Irrigation Controller Fact Sheet

    EPA Pesticide Factsheets

    WaterSense labeled irrigation controllers, which act like a thermostat for your sprinkler system by telling it when to turn on and off, use local weather and landscape conditions to tailor watering schedules to actual conditions on the site.

  15. Method for producing labeled single-stranded nucleic acid probes

    DOEpatents

    Dunn, John J.; Quesada, Mark A.; Randesi, Matthew

    1999-10-19

    Disclosed is a method for the introduction of unidirectional deletions in a cloned DNA segment. More specifically, the method comprises providing a recombinant DNA construct comprising a DNA segment of interest inserted in a cloning vector, the cloning vector having an f1 endonuclease recognition sequence adjacent to the insertion site of the DNA segment of interest. The recombinant DNA construct is then contacted with the protein pII encoded by gene II of phage f1 thereby generating a single-stranded nick. The nicked DNA is then contacted with E. coli Exonuclease III thereby expanding the single-stranded nick into a single-stranded gap. The single-stranded gapped DNA is then contacted with a single-strand-specific endonuclease thereby producing a linearized DNA molecule containing a double-stranded deletion corresponding in size to the single-stranded gap. The DNA treated in this manner is then incubated with DNA ligase under conditions appropriate for ligation. Also disclosed is a method for producing single-stranded DNA probes. In this embodiment, single-stranded gapped DNA, produced as described above, is contacted with a DNA polymerase in the presence of labeled nucleotides to fill in the gap. This DNA is then linearized by digestion with a restriction enzyme which cuts outside the DNA segment of interest. The product of this digestion is then denatured to produce a labeled single-stranded nucleic acid probe.

  16. Spectroscopic Methods for Label-Free Optical Nanoscopy

    NASA Astrophysics Data System (ADS)

    Chandler, John E.

    It is becoming increasingly evident that the nanoscale organization and structure of macromolecules play a significant role in determining the function and properties of biological systems. To understand the relationships between biological structure and function at nanometer length scales, there is a need for methods which enable imaging of intact nanoscale biological structure. An ideal technique for these applications is sensitive to nanoscale structure below the resolution limit of conventional optical microscopy ( 200 nm), achieves label-free contrast, is non-perturbing to biological samples, is quantitative, is capable of molecular specificity, is high-throughput, and finally is simple, enabling widespread utilization. Existing techniques meet some of these criteria, but all have limitations. For example, super-resolution optical microscopy methods achieve molecular-specific nanoscale resolution well below the resolution limit of conventional optical microscopes, however, they rely on fluorescent labels often at high densities that can be toxic and can often require potentially damaging illumination intensities for imaging. As a result, there remains a need for label-free optical techniques to study the nanoscale structural properties of cells. To address this need, the development of instrumentation and algorithms for Partial Wave Spectroscopic (PWS) microscopy will be described. PWS is a spectroscopic, label-free, nanoscale sensitive microscope which, senses rather than resolves structure below the resolution limit of conventional microscopes ( 200nm). First, PWS has shown utility as a diagnostic screening tool for cancer due to nanoscale structural alterations that occur in cells as part of the earliest stages of carcinogenesis. Instrumentation and algorithms developed to enable high-throughput cancer screening applications will be described. Further enhancement of data acquisition and analysis speed will then be described through the development of new

  17. Measurement of vascular water transport in human subjects using time-resolved pulsed arterial spin labelling.

    PubMed

    Bibic, Adnan; Knutsson, Linda; Schmidt, Anders; Henningsson, Erik; Månsson, Sven; Abul-Kasim, Kasim; Åkeson, Jonas; Gunther, Matthias; Ståhlberg, Freddy; Wirestam, Ronnie

    2015-08-01

    Most approaches to arterial spin labelling (ASL) data analysis aim to provide a quantitative measure of the cerebral blood flow (CBF). This study, however, focuses on the measurement of the transfer time of blood water through the capillaries to the parenchyma (referred to as the capillary transfer time, CTT) as an alternative parameter to characterise the haemodynamics of the system. The method employed is based on a non-compartmental model, and no measurements need to be added to a common time-resolved ASL experiment. Brownian motion of labelled spins in a potential was described by a one-dimensional general Langevin equation as the starting point, and as a Fokker-Planck differential equation for the averaged distribution of labelled spins at the end point, which takes into account the effects of flow and dispersion of labelled water by the pseudorandom nature of the microvasculature and the transcapillary permeability. Multi-inversion time (multi-TI) ASL data were acquired in 14 healthy subjects on two occasions in a test-retest design, using a pulsed ASL sequence and three-dimensional gradient and spin echo (3D-GRASE) readout. Based on an error analysis to predict the size of a region of interest (ROI) required to obtain reasonably precise parameter estimates, data were analysed in two relatively large ROIs, i.e. the occipital lobe (OC) and the insular cortex (IC). The average values of CTT in OC were 260 ± 60 ms in the first experiment and 270 ± 60 ms in the second experiment. The corresponding IC values were 460 ± 130 ms and 420 ± 139 ms, respectively. Information related to the water transfer time may be important for diagnostics and follow-up of cerebral conditions or diseases characterised by a disrupted blood-brain barrier or disturbed capillary blood flow.

  18. Combinatorial Labeling Method for Improving Peptide Fragmentation in Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Kuchibhotla, Bhanuramanand; Kola, Sankara Rao; Medicherla, Jagannadham V.; Cherukuvada, Swamy V.; Dhople, Vishnu M.; Nalam, Madhusudhana Rao

    2017-06-01

    Annotation of peptide sequence from tandem mass spectra constitutes the central step of mass spectrometry-based proteomics. Peptide mass spectra are obtained upon gas-phase fragmentation. Identification of the protein from a set of experimental peptide spectral matches is usually referred as protein inference. Occurrence and intensity of these fragment ions in the MS/MS spectra are dependent on many factors such as amino acid composition, peptide basicity, activation mode, protease, etc. Particularly, chemical derivatizations of peptides were known to alter their fragmentation. In this study, the influence of acetylation, guanidinylation, and their combination on peptide fragmentation was assessed initially on a lipase (LipA) from Bacillus subtilis followed by a bovine six protein mix digest. The dual modification resulted in improved fragment ion occurrence and intensity changes, and this resulted in the equivalent representation of b- and y-type fragment ions in an ion trap MS/MS spectrum. The improved representation has allowed us to accurately annotate the peptide sequences de novo. Dual labeling has significantly reduced the false positive protein identifications in standard bovine six peptide digest. Our study suggests that the combinatorial labeling of peptides is a useful method to validate protein identifications for high confidence protein inference. [Figure not available: see fulltext.

  19. Assay for vitamin B12 absorption and method of making labeled vitamin B12

    DOEpatents

    Anderson, Peter J [Davis, CA; Dueker, Stephen [Davis, CA; Miller, Joshua [Davis, CA; Green, Ralph [Elmacero, CA; Roth, John [Davis, CA; Carkeet, Colleen [Silver Spring, MD; Buchholz,; Bruce, A [Orinda, CA

    2012-06-19

    The invention provides methods for labeling vitamin B12 with .sup.14C, .sup.13C, tritium, and deuterium. When radioisotopes are used, the invention provides for methods of labeling B12 with high specific activity. The invention also provides labeled vitamin B12 compositions made in accordance with the invention.

  20. Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels

    DOEpatents

    Mathies, Richard A.; Singhal, Pankaj; Xie, Jin; Glazer, Alexander N.

    2002-01-01

    This invention relates to a microfabricated capillary electrophoresis chip for detecting multiple redox-active labels simultaneously using a matrix coding scheme and to a method of selectively labeling analytes for simultaneous electrochemical detection of multiple label-analyte conjugates after electrophoretic or chromatographic separation.

  1. Diffusion of oxygen in water and hydrocarbons using an electron spin resonance spin-label technique.

    PubMed Central

    Subczynski, W K; Hyde, J S

    1984-01-01

    The Smoluchowski equation for the bimolecular collision rate of dissolved oxygen molecules with spin labels yielded values for the diffusion constant of oxygen in water that are in agreement with the Stokes-Einstein equation (D infinity T/eta, where eta is the macroscopic viscosity) and with published values obtained by conventional methods. Heisenberg exchange at an interaction distance of 4.5 A occurs with a probability close to one for each encounter. In mixed hydrocarbons (olive oil, paraffin oils) and sec-butyl benzene, D infinity (T/eta)rho, where rho lies between 0.5 and 1. Oxygen diffuses in the hydrocarbons between 10 and 100 times more rapidly than predicted from the macroscopic viscosity. Similar results would be expected for diffusion of oxygen in model and biological membranes. Parallel measurements of rotational diffusion of the spin labels show little correlation with measurements of translational diffusion of oxygen. Dipolar interactions between spin labels and oxygen appear negligible except in the limit of highest viscosities. PMID:6326877

  2. Multispot, label-free biodetection at a phantom plastic-water interface.

    PubMed

    Giavazzi, Fabio; Salina, Matteo; Cerbino, Roberto; Bassi, Mattia; Prosperi, Davide; Ceccarello, Erica; Damin, Francesco; Sola, Laura; Rusnati, Marco; Chiari, Marcella; Chini, Bice; Bellini, Tommaso; Buscaglia, Marco

    2013-06-04

    Recognizing and quantifying specific biomolecules in aqueous samples are constantly needed in research and diagnostic laboratories. As the typical detection procedures are rather lengthy and involve the use of labeled secondary antibodies or other agents to provide a signal, efforts have been made over the last 10 y to develop alternative label-free methods that enable direct detection. We propose and demonstrate an extremely simple, low-cost, label-free biodetector based on measuring the intensity of light reflected by the interface between a fluid sample and an amorphous fluoropolymer substrate having a refractive index very close to that of water and hosting various antibodies immobilized in spots. Under these index-matching conditions, the amount of light reflected by the interface allows straightforward quantification of the amount of antigen binding to each spot. Using antibodies targeting heterologous immunoglobulins and antigens commonly used as markers for diagnoses of hepatitis B and HIV, we demonstrate the limit of detection of a few picograms per square millimeter of surface-bound molecules. We also show that direct and real-time access to the amount of binding molecules allows the precise extrapolation of adhesion rates, from which the concentrations of antigens in solution can be estimated down to fractions of nanograms per milliliter.

  3. Multispot, label-free biodetection at a phantom plastic–water interface

    PubMed Central

    Giavazzi, Fabio; Salina, Matteo; Cerbino, Roberto; Bassi, Mattia; Prosperi, Davide; Ceccarello, Erica; Damin, Francesco; Sola, Laura; Rusnati, Marco; Chiari, Marcella; Chini, Bice; Bellini, Tommaso; Buscaglia, Marco

    2013-01-01

    Recognizing and quantifying specific biomolecules in aqueous samples are constantly needed in research and diagnostic laboratories. As the typical detection procedures are rather lengthy and involve the use of labeled secondary antibodies or other agents to provide a signal, efforts have been made over the last 10 y to develop alternative label-free methods that enable direct detection. We propose and demonstrate an extremely simple, low-cost, label-free biodetector based on measuring the intensity of light reflected by the interface between a fluid sample and an amorphous fluoropolymer substrate having a refractive index very close to that of water and hosting various antibodies immobilized in spots. Under these index-matching conditions, the amount of light reflected by the interface allows straightforward quantification of the amount of antigen binding to each spot. Using antibodies targeting heterologous immunoglobulins and antigens commonly used as markers for diagnoses of hepatitis B and HIV, we demonstrate the limit of detection of a few picograms per square millimeter of surface-bound molecules. We also show that direct and real-time access to the amount of binding molecules allows the precise extrapolation of adhesion rates, from which the concentrations of antigens in solution can be estimated down to fractions of nanograms per milliliter. PMID:23696673

  4. Measurement of protein synthesis using heavy water labeling and peptide mass spectrometry: Discrimination between major histocompatibility complex allotypes

    PubMed Central

    De Riva, Alessandra; Deery, Michael J.; McDonald, Sarah; Lund, Torben; Busch, Robert

    2010-01-01

    Methodological limitations have hampered the use of heavy water (2H2O), a convenient, universal biosynthetic label, for measuring protein synthesis. Analyses of 2H-labeled amino acids are sensitive to contamination; labeling of peptides has been measured for a few serum proteins, but this approach awaits full validation. Here we describe a method for quantifying protein synthesis by peptide mass spectrometry (MS) after 2H2O labeling, as applied to various proteins of the major histocompatibility complex (MHC). Human and murine antigen-presenting cells were cultured in medium containing 5% 2H2O; class I and class II MHC proteins were immunoprecipitated, bands were excised, and Ala-/Gly-rich, allele-specific tryptic peptides were identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). Mass isotopomer distributions were quantified precisely by LC–MS and shifted markedly on 2H2O labeling. Experimental data agreed closely with models obtained by mass isotopomer distribution analysis (MIDA) and were consistent with contributions from Ala, Gly, and other amino acids to labeling. Estimates of fractional protein synthesis from peptides of the same protein were precise and internally consistent. The method was capable of discriminating between MHC isotypes and alleles, applicable to primary cells, and readily extendable to other proteins. It simplifies measurements of protein synthesis, enabling novel applications in physiology, in genotype/phenotype interactions, and potentially in kinetic proteomics. PMID:20406617

  5. Validation of a non-invasive blood-sampling technique for doubly-labelled water experiments.

    PubMed

    Voigt, Christian C; Helversen, Otto Von; Michener, Robert H; Kunz, Thomas H

    2003-04-01

    Two techniques for bleeding small mammals have been used in doubly-labeled water (DLW) studies, including vena puncture and the use of starved nymphal stages of hematophagous reduviid bugs (Reduviidae, Hemiptera). In this study, we tested the validity of using reduviid bugs in doubly-labeled water experiments. We found that the isotope enrichment in initial blood samples collected with bugs was significantly lower compared to isotope enrichment in blood samples obtained using vena puncture. We therefore used the desiccation method for estimating total body water (TBW) in DLW experiments because TBW calculated using the isotope dilution method was overestimated when blood samples were collected using reduviid bugs. In our validation experiment with nectar-feeding bats (Glossophaga soricina), we compared estimates of daily energy expenditure (DEE) using DLW with those derived from the energy balance method. We considered Speakman's equation (controlling for 25% fractionated water loss) as the most appropriate for our study animal and calculated DEE accordingly. On average, DEE estimated with DLW was not significantly different from the mean value obtained with the energy balance method (mean deviation 1.2%). We conclude that although bug hemolymph or intestinal liquids most likely contaminate the samples, estimates of DEE are still valid because the DLW method does not depend on absolute isotope enrichments but on the rate of isotope decrease over time. However, dilution of blood with intestinal liquids or hemolymph from a bug may lead to larger variation in DEE estimates. We also tested how the relative error of DLW estimates changed with varying assumptions about fractionation. We used three additional equations for calculating DEE in DLW experiments. The basic equation for DLW experiments published by Lifson and McClintock (LM-6) assumes no fractionation, resulted in an overestimate of DEE by 10%. Nagy's equation (N-2) controls for changes in body mass but not for

  6. Foraging flights of the white-tailed tropicbird (Phaethon lepturus): Radiotracking and doubly-labelled water

    USGS Publications Warehouse

    Pennycuick, C.J.; Shaffner, F.C.; Fuller, M.R.; Obrecht, H.H.; Sternberg, L.

    1990-01-01

    Radiotracking transmitters were fitted to White-tailed Tropicbirds nesting at Culebra, Puerto Rico. Foragers were located by light aircraft out to 89 km SSW of the nesting colony, over a deep-water foraging area south of Vieques Island, Puerto Rico and west of St Croix, U. S. Virgin Islands. Two birds were followed out to 176 km NNW from the colony, over the Puerto Rico Trench, but these did not subsequently return. Foragers carrying radio transmitters performed similarly to those without, in terms of duration of absence from the colony, and mass of food brought for the chick. However, measuremetns of energy consumption by the doubly labelled water method indicated that birds with transmitters consumed significantly more energy than those without.

  7. Interdisciplinary Methods in Water Resources

    ERIC Educational Resources Information Center

    Cosens, Barbara; Fiedler, Fritz; Boll, Jan; Higgins, Lorie; Johnson, Gary; Kennedy, Brian; Strand, Eva; Wilson, Patrick; Laflin, Maureen

    2011-01-01

    In the face of a myriad of complex water resource issues, traditional disciplinary separation is ineffective in developing approaches to promote a sustainable water future. As part of a new graduate program in water resources, faculty at the University of Idaho have developed a course on interdisciplinary methods designed to prepare students for…

  8. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... all times during the marketing of a fur product the required label shall have a minimum dimension of... secure manner and with sufficient permanency to remain thereon throughout the sale, resale,...

  9. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... all times during the marketing of a fur product the required label shall have a minimum dimension of... secure manner and with sufficient permanency to remain thereon throughout the sale, resale,...

  10. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... all times during the marketing of a fur product the required label shall have a minimum dimension of... secure manner and with sufficient permanency to remain thereon throughout the sale, resale,...

  11. Simple SPION Incubation as an Efficient Intracellular Labeling Method for Tracking Neural Progenitor Cells Using MRI

    PubMed Central

    D. M., Jayaseema; Lai, Jiann-Shiun; Hueng, Dueng-Yuan; Chang, Chen

    2013-01-01

    Cellular magnetic resonance imaging (MRI) has been well-established for tracking neural progenitor cells (NPC). Superparamagnetic iron oxide nanoparticles (SPIONs) approved for clinical application are the most common agents used for labeling. Conventionally, transfection agents (TAs) were added with SPIONs to facilitate cell labeling because SPIONs in the native unmodified form were deemed inefficient for intracellular labeling. However, compelling evidence also shows that simple SPION incubation is not invariably ineffective. The labeling efficiency can be improved by prolonged incubation and elevated iron doses. The goal of the present study was to establish simple SPION incubation as an efficient intracellular labeling method. To this end, NPCs derived from the neonatal subventricular zone were incubated with SPIONs (Feridex®) and then evaluated in vitro with regard to the labeling efficiency and biological functions. The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake. Evidence from light microscopy, electron microscopy, chemical analysis, and magnetic resonance imaging confirmed the effectiveness of the labeling. Additionally, biological assays showed that the labeled NPCs exhibited unaffected viability, oxidative stress, apoptosis and differentiation. In the demonstrated in vivo cellular MRI experiment, the hypointensities representing the SPION labeled NPCs remained observable throughout the entire tracking period. The findings indicate that simple SPION incubation without the addition of TAs is an efficient intracellular magnetic labeling method. This simple approach may be considered as an alternative approach to the mainstream labeling method that involves the use of TAs. PMID:23468856

  12. Simple SPION incubation as an efficient intracellular labeling method for tracking neural progenitor cells using MRI.

    PubMed

    Chen, Chiao-Chi V; Ku, Min-Chi; D M, Jayaseema; Lai, Jiann-Shiun; Hueng, Dueng-Yuan; Chang, Chen

    2013-01-01

    Cellular magnetic resonance imaging (MRI) has been well-established for tracking neural progenitor cells (NPC). Superparamagnetic iron oxide nanoparticles (SPIONs) approved for clinical application are the most common agents used for labeling. Conventionally, transfection agents (TAs) were added with SPIONs to facilitate cell labeling because SPIONs in the native unmodified form were deemed inefficient for intracellular labeling. However, compelling evidence also shows that simple SPION incubation is not invariably ineffective. The labeling efficiency can be improved by prolonged incubation and elevated iron doses. The goal of the present study was to establish simple SPION incubation as an efficient intracellular labeling method. To this end, NPCs derived from the neonatal subventricular zone were incubated with SPIONs (Feridex®) and then evaluated in vitro with regard to the labeling efficiency and biological functions. The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake. Evidence from light microscopy, electron microscopy, chemical analysis, and magnetic resonance imaging confirmed the effectiveness of the labeling. Additionally, biological assays showed that the labeled NPCs exhibited unaffected viability, oxidative stress, apoptosis and differentiation. In the demonstrated in vivo cellular MRI experiment, the hypointensities representing the SPION labeled NPCs remained observable throughout the entire tracking period. The findings indicate that simple SPION incubation without the addition of TAs is an efficient intracellular magnetic labeling method. This simple approach may be considered as an alternative approach to the mainstream labeling method that involves the use of TAs.

  13. Comparison of energy estimates in chronic kidney disease using doubly-labelled water.

    PubMed

    Sridharan, S; Wong, J; Vilar, E; Farrington, K

    2016-02-01

    Total energy expenditure (TEE) is estimated in clinical practice as a combined measure of resting energy expenditure and physical activity level. Commonly available questionnaires to estimate physical activity level have not been validated in patients with kidney disease using the doubly-labelled water method. This prospective, cross-sectional study was conducted on 40 patients with chronic kidney disease stages 1-5 with the objective of validating two physical activity questionnaires: the Recent Physical Activity Questionnaire (RPAQ) and the Stanford 7-day recall questionnaire. TEE was measured using doubly-labelled water technique. TEE was also estimated using predicted resting energy expenditure and estimated physical activity measures from the questionnaires. Measured TEE correlated better with TEE estimated from RPAQ compared to that from the Stanford questionnaire. In Bland-Altman analysis, TEE estimated from RPAQ had the least bias and narrower limits of agreement compared to the measured TEE. A metabolic equivalent of task value of 1.3 for the unaccounted time in RPAQ provided the best approximation of estimated TEE to the measured TEE. RPAQ is an acceptable questionnaire tool for assessing physical activity level in patients with chronic kidney disease. © 2015 The British Dietetic Association Ltd.

  14. PRN 93-8: Labeling Statement Prohibiting Application to Water; Amendment to PR Notice 93-3

    EPA Pesticide Factsheets

    This notice adds one paragraph to P.R. Notice 93-3, which requested registrants to amend product labeling to include a new labeling statement prohibiting application of a pesticide to water for certain products.

  15. Simple, rapid method for the preparation of isotopically labeled formaldehyde

    SciTech Connect

    Hooker, Jacob Matthew; Schonberger, Matthias; Schieferstein, Hanno; Fowler, Joanna S.

    2011-10-04

    Isotopically labeled formaldehyde (*C.sup..sctn.H.sub.2O) is prepared from labeled methyl iodide (*C.sup..sctn.H.sub.3I) by reaction with an oxygen nucleophile having a pendant leaving group. The mild and efficient reaction conditions result in good yields of *C.sup..sctn.H.sub.2O with little or no *C isotopic dilution. The simple, efficient production of .sup.11CH.sub.2O is described. The use of the .sup.11CH.sub.2O for the formation of positron emission tomography tracer compounds is described. The reaction can be incorporated into automated equipment available to radiochemistry laboratories. The isotopically labeled formaldehyde can be used in a variety of reactions to provide radiotracer compounds for imaging studies as well as for scintillation counting and autoradiography.

  16. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... CONGRESS RULE CONCERNING DISCLOSURES REGARDING ENERGY CONSUMPTION AND WATER USE OF CERTAIN HOME APPLIANCES AND OTHER PRODUCTS REQUIRED UNDER THE ENERGY POLICY AND CONSERVATION ACT (âAPPLIANCE LABELING RULEâ..., clothes washers, and water heaters must contain the model's estimated annual energy consumption as...

  17. 40 CFR 600.115-11 - Criteria for determining the fuel economy label calculation method.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... VEHICLES Fuel Economy and Carbon-Related Exhaust Emission Test Procedures § 600.115-11 Criteria for... derived 5-cycle method for determining fuel economy label values, as specified in § 600.210-08(a)(2) or (b... economy label values must be determined according to the vehicle-specific 5-cycle method specified...

  18. On multilabel classification methods of incompletely labeled biomedical text data.

    PubMed

    Kolesov, Anton; Kamyshenkov, Dmitry; Litovchenko, Maria; Smekalova, Elena; Golovizin, Alexey; Zhavoronkov, Alex

    2014-01-01

    Multilabel classification is often hindered by incompletely labeled training datasets; for some items of such dataset (or even for all of them) some labels may be omitted. In this case, we cannot know if any item is labeled fully and correctly. When we train a classifier directly on incompletely labeled dataset, it performs ineffectively. To overcome the problem, we added an extra step, training set modification, before training a classifier. In this paper, we try two algorithms for training set modification: weighted k-nearest neighbor (WkNN) and soft supervised learning (SoftSL). Both of these approaches are based on similarity measurements between data vectors. We performed the experiments on AgingPortfolio (text dataset) and then rechecked on the Yeast (nontext genetic data). We tried SVM and RF classifiers for the original datasets and then for the modified ones. For each dataset, our experiments demonstrated that both classification algorithms performed considerably better when preceded by the training set modification step.

  19. A Novel Method for Assigning R, S Labels to Enantiomers.

    ERIC Educational Resources Information Center

    Huheey, James E.

    1986-01-01

    Discusses ways of teaching students about how to assign R (rectus) and S (sinister) labels to enantiomers by using their hands as models. The chirality of the human hands follows the Cahn-Ingold-Prelog Rules for assigning enantiomers and infers the correct chirality of molecules shown in two-dimensional drawings. (TW)

  20. A Novel Method for Assigning R, S Labels to Enantiomers.

    ERIC Educational Resources Information Center

    Huheey, James E.

    1986-01-01

    Discusses ways of teaching students about how to assign R (rectus) and S (sinister) labels to enantiomers by using their hands as models. The chirality of the human hands follows the Cahn-Ingold-Prelog Rules for assigning enantiomers and infers the correct chirality of molecules shown in two-dimensional drawings. (TW)

  1. Clean Water Act Analytical Methods

    EPA Pesticide Factsheets

    EPA publishes laboratory analytical methods (test procedures) that are used by industries and municipalities to analyze the chemical, physical and biological components of wastewater and other environmental samples required by the Clean Water Act.

  2. Co-Permeability of 3H-Labeled Water and 14C-Labeled Organic Acids across Isolated Plant Cuticles1

    PubMed Central

    Niederl, Sabine; Kirsch, Thomas; Riederer, Markus; Schreiber, Lukas

    1998-01-01

    Penetration of 3H-labeled water (3H2O) and the 14C-labeled organic acids benzoic acid ([14C]BA), salicylic acid ([14C]SA), and 2,4-dichlorophenoxyacetic acid ([14C]2,4-D) were measured simultaneously in isolated cuticular membranes of Prunus laurocerasus L., Ginkgo biloba L., and Juglans regia L. For each of the three pairs of compounds (3H2O/[14C]BA, 3H2O/[14C]SA, and 3H2O/[14C]2,4-D) rates of cuticular water penetration were highly correlated with the rates of penetration of the organic acids. Therefore, water and organic acids penetrated the cuticles by the same routes. With the combination 3H2O/[14C]BA, co-permeability was measured with isolated cuticles of nine other plant species. Permeances of 3H2O of all 12 investigated species were highly correlated with the permeances of [14C]BA (r2 = 0.95). Thus, cuticular transpiration can be predicted from BA permeance. The application of this experimental method, together with the established prediction equation, offers the opportunity to answer several important questions about cuticular transport physiology in future investigations.

  3. Methods for labeling .beta.-amyloid plaques and neurofibrillary tangles

    DOEpatents

    Barrio, Jorge R.; Petric, Andrej; Satyamurthy, Nagichettiar; Small, Gary W.; Cole, Gregory M.; Huang, Sung-Cheng

    2001-01-01

    A method for labeling .beta.-amyloid plaques and neurofibrillary tangles in vivo and in vitro, comprises contacting a compound of formula (I): ##STR1## with mammalian tissue. In formula (I), R.sub.1 is selected from the group consisting of --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C.dbd.C(CN).sub.2 -alkyl, --C.dbd.C(CN).sub.2 -alkylenyl-R.sub.4 , ##STR2## R.sub.4 is a radical selected from the group consisting of alkyl, substituted alkyl, aryl and substituted aryl; R.sub.5, is a radical selected from the group consisting of --NH.sub.2, --OH, --SH, --NH-alkyl, --NHR.sub.4, --NH-alkylenyl-R.sub.4, --O-alkyl, --O-alkylenyl-R.sub.4, --S-alkyl, and --S-alkylenyl-R.sub.4 ; R.sub.6 is a radical selected from the group consisting of --CN, --COOH, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)-halogen, --C(O)NH , --C(O)NH-alkyl, --C(O)NH-alkylenyl-R.sub.4 ; R.sub.7 is a radical selected from the group consisting of O, NH, and S; and R.sub.8 is N, O or S. R.sub.2 and R.sub.3 are each independently selected from the group consisting of alkyl and alkylenyl-R.sub.10, wherein R.sub.10 is selected from the group consisting of --OH, --OTs, halogen, spiperone, spiperone ketal and spiperone-3-yl. Alternatively, R.sub.2 and R.sub.3 together form a heterocyclic ring, optionally substituted with at least one radical selected from the group consisting of alkyl, alkoxy, OH, OTs, halogen, alkylenyl-R.sub.10, carbonyl, spiperone, spiperone ketal and spiperone-3-yl. In the compounds of formula (I), one or more of the hydrogen, halogen or carbon atoms can, optionally, be replaced with a radiolabel.

  4. Methods for labeling .beta.-amyloid plaques and neurofibrillary tangles

    DOEpatents

    Barrio, Jorge R.; Petric, Andrej; Satyamurthy, Nagichettiar; Small, Gary W.; Cole, Gregory M.; Huang, Sung-Cheng

    2003-12-09

    A method for labeling .beta.-amyloid plaques and neurofibrillary tangles in vivo and in vitro, comprises contacting a compound of formula (I): ##STR1## with mammalian tissue. In formula (I), R.sub.1 is selected from the group consisting of --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C.dbd.C(CN).sub.2 -alkyl, --C.dbd.C(CN).sub.2 -alkylenyl-R.sub.4, ##STR2## R.sub.4 is a radical selected from the group consisting of alkyl, substituted alkyl, aryl and substituted aryl; R.sub.5 is a radical selected from the group consisting of --NH.sub.2, --OH, --SH, --NH-alkyl, --NHR.sub.4, --NH-alkylenyl-R.sub.4, --O-alkyl, --O-alkylenyl-R.sub.4, --S-alkyl, and --S-alkylenyl-R.sub.4 ; R.sub.6 is a radical selected from the group consisting of --CN, --COOH, --C(O)O-alkyl, --C(O)O-alkylenyl-R.sub.4, --C(O)-alkyl, --C(O)-alkylenyl-R.sub.4, --C(O)-halogen, --C(O)NH, --C(O)NH-alkyl, --C(O)NH-alkylenyl-R.sub.4 ; R.sub.7 is a radical selected from the group consisting of O, NH, and S; and R.sub.8 is N, O or S. R.sub.2 and R.sub.3 are each independently selected from the group consisting of alkyl and alkylenyl-R.sub.10, wherein R.sub.10 is selected from the group consisting of --OH, --OTs, halogen, spiperone, spiperone ketal and spiperone-3-yl. Alternatively, R.sub.2 and R.sub.3 together form a heterocyclic ring, optionally substituted with at least one radical selected from the group consisting of alkyl, alkoxy, OH, OTs, halogen, alkylenyl-R.sub.10, carbonyl, spiperone, spiperone ketal and spiperone-3-yl. In the compounds of formula (I), one or more of the hydrogen, halogen or carbon atoms can, optionally, be replaced with a radiolabel.

  5. Novel image processing method study for a label-free optical biosensor

    NASA Astrophysics Data System (ADS)

    Yang, Chenhao; Wei, Li'an; Yang, Rusong; Feng, Ying

    2015-10-01

    Optical biosensor is generally divided into labeled type and label-free type, the former mainly contains fluorescence labeled method and radioactive-labeled method, while fluorescence-labeled method is more mature in the application. The mainly image processing methods of fluorescent-labeled biosensor includes smooth filtering, artificial gridding and constant thresholding. Since some fluorescent molecules may influence the biological reaction, label-free methods have been the main developing direction of optical biosensors nowadays. The using of wider field of view and larger angle of incidence light path which could effectively improve the sensitivity of the label-free biosensor also brought more difficulties in image processing, comparing with the fluorescent-labeled biosensor. Otsu's method is widely applied in machine vision, etc, which choose the threshold to minimize the intraclass variance of the thresholded black and white pixels. It's capacity-constrained with the asymmetrical distribution of images as a global threshold segmentation. In order to solve the irregularity of light intensity on the transducer, we improved the algorithm. In this paper, we present a new image processing algorithm based on a reflectance modulation biosensor platform, which mainly comprises the design of sliding normalization algorithm for image rectification and utilizing the improved otsu's method for image segmentation, in order to implement automatic recognition of target areas. Finally we used adaptive gridding method extracting the target parameters for analysis. Those methods could improve the efficiency of image processing, reduce human intervention, enhance the reliability of experiments and laid the foundation for the realization of high throughput of label-free optical biosensors.

  6. Parallel assessment of nutrition and activity in athletes: validation against doubly labelled water, 24-h urea excretion, and indirect calorimetry.

    PubMed

    Koehler, Karsten; Braun, Hans; De Marees, Markus; Fusch, Gerhard; Fusch, Christoph; Mester, Joachim; Schaenzer, Wilhelm

    2010-11-01

    The assessment of nutrition and activity in athletes requires accurate and precise methods. The aim of this study was to validate a protocol for parallel assessment of diet and exercise against doubly labelled water, 24-h urea excretion, and respiratory gas exchange. The participants were 14 male triathletes under normal training conditions. Energy intake and doubly labelled water were weakly associated with each other (r = 0.69, standard error of estimate [SEE] = 304 kcal x day(-1)). Protein intake was strongly correlated with 24-h urea (r = 0.89) but showed considerable individual variation (SEE = 0.34 g kg(-1) x day(-1)). Total energy expenditure based on recorded activities was highly correlated with doubly labelled water (r = 0.95, SEE = 195 kcal x day(-1)) but was proportionally biased. During running and cycling, estimated exercise energy expenditure was highly correlated with gas exchange (running: r = 0.89, SEE = 1.6 kcal x min(-1); cycling: r = 0.95, SEE = 1.4 kcal x min(-1)). High exercise energy expenditure was slightly underestimated during running. For nutrition data, variations appear too large for precise measurements in individual athletes, which is a common problem of dietary assessment methods. Despite the high correlations of total energy expenditure and exercise energy expenditure with reference methods, a correction for systematic errors is necessary for the valid estimation of energetic requirements in individual athletes.

  7. Lanthanide Label Array Method for Identification and Adulteration of Honey and Cacao.

    PubMed

    Härmä, Harri; Peltomaa, Riikka; Pihlasalo, Sari

    2015-07-07

    A generic, cost-effective, and simple method has been developed to fingerprint liquids to differentiate food brands and ingredients. The method is based on a label array using nonspecific long lifetime unstable luminescent lanthanide labels. The interaction between the liquid sample and the label is typically detrimental to the luminescence of the unstable chelate leading to a sample-dependent luminescence-intensity array. The label-array method is a unique approach as the array of unstable chelates is extremely inexpensive to produce and possesses high sensitivity due to spectral as well as unstable structural properties of the lanthanide label. The global method has been applied to distinguish commercial honey and cacao brands to demonstrate its feasibility as honey and cacao are among the most adulterated food products.

  8. Blood cell labelling. Theory and methods: radiation hazards.

    PubMed

    Trott, N G; Akbari, R B

    1984-02-03

    The chief physical properties of the radionuclide In111 are outlined, and compared with those of three other radionuclides, Tc99m, I131 and Cr51 which have similar applications. It is pointed out that the gamma-rays of In111 are appreciably more penetrating in lead than those of Tc99m and the significance of this, both in the use of shielding on syringes and in the effectiveness of lead glass screens is discussed. Examples are given of the dosimetry for In111 labelled cells in humans and it is noted that the absorbed dose in the spleen per mCi (37 MBq) injected may be some 10 rad (0.1 Gy). The problems that have been noted of damage to cells arising from oxine labelling and now considered to be due to radiation damage are briefly reviewed.

  9. Intrinsic mineral labeling of edible plants: methods and uses

    SciTech Connect

    Weaver, C.M.

    1985-01-01

    The fate of minerals can be conveniently studied through intrinsic labeling techniques. The mineral of interest is biologically incorporated into the food in a form that can be distinguished analytically from the natural form of the element. Radiolabels have traditionally been used to study such problems as the uptake of minerals by plants, the gross and subcellular mineral distribution in plant tissues, the form and associations of the deposited mineral, and the bioavailability of minerals to animals and humans. The use of stable (nonradioactive) isotopes as a label offers the potential of safely studying bioavailability of minerals from individual foods in human population groups of all ages using foods processed in normal food handling and processing facilities. 114 references.

  10. Methods for nanoparticle labeling of ricin and effect on toxicity

    NASA Astrophysics Data System (ADS)

    Wark, Alastair W.; Yu, Jun; Lindsay, Christopher D.; Nativo, Paola; Graham, Duncan

    2009-09-01

    The unique optical properties associated with nanostructured materials that support the excitation of surface plasmons offer many new opportunities for the enhanced optical investigation of biological materials that pose a security threat. In particular, ricin is considered a significant bioterrorism risk due to its high toxicity combined with its ready availability as a byproduct in castor oil production. Therefore, the development of optical techniques capable of rapid on-site toxin detection with high molecular specificity and sensitivity continues to be of significant importance. Furthermore, understanding of the ricin cell entry and intracellular pathways remains poor due to a lack of suitable bioanalytical techniques. Initial work aimed at simultaneously tackling both these issues is described where different approaches for the nanoparticle labeling of ricin are investigated along with changes in ricin toxicity associated with the labeling process.

  11. Water-soluble phosphines for direct labeling of peptides with technetium and rhenium: insights from electrospray mass spectrometry.

    PubMed

    Greenland, William E P; Blower, Philip J

    2005-01-01

    Direct labeling of salmon calcitonin (sCT) is possible in one step using water-soluble phosphines (sulfonated triphenylphosphines) as the reducing agent both for disulfide and for pertechnetate. Phosphines were the most efficient reducing agent for disulfide bonds among those examined. The phosphines both reduced the pertechnetate to Tc(III), and contributed to the technetium coordination sphere in the labeled product. In contrast, the phosphines did not reduce rhenium below oxidation state V, nor did they participate in the rhenium coordination sphere in the labeled peptide. Instead, the expected oxorhenium(V) moiety was incorporated. Both Tc and Re labeling processes gave rise to dimers with two peptides linked by the metal center, as well as simple monomeric species. Positive mode electrospray mass spectrometry not only revealed the presence of phosphine bound to technetium and oxygen bound to rhenium in the metallopeptides but also revealed the oxidation states of the metals. Electrospray mass spectrometry is proving to be an exceptionally valuable technique for characterizing radiopharmaceuticals. Although the one-step direct labeling method described gives mixed products and poor receptor affinity when applied to the small peptide sCT, it might be readily adapted to monoclonal antibodies.

  12. New method for covalent fluorescent biomolecule labeling with hemicyanine dye.

    PubMed

    Kostenko, Olexander M; Kovalska, Vladyslava B; Volkova, Kateryna D; Shaytanov, Pavel; Kocheshev, Igor O; Slominskiy, Yuriy L; Pisareva, Irina V; Yarmoluk, Sergiy M

    2006-07-01

    Fluorescent chromophore, alkylamino-(tetra-hydronaphthalenylidene)- benzothiazolium derivatives (HBTN dyes), are proposed as covalent labels for proteins via aliphatic amino groups. Spectral-luminescent properties of 3-methyl-2-{(E)-[7-(methylamino)-4,4a,5,6-tetra-hydronaphthalen-2(3H)-ylidene]methyl}-1,3-benzothiazol-3-ium chloride (HBTN, R=Me) and its predecessor, 2-[(E)-(7-methoxy-4,4a,5,6-tetrahydronaphthalen-2(3H)-ylidene)methyl]-3-methyl-1,3-benzothiazol-3-ium chloride (ABTN), are studied for free dyes and in the presence of DNA and BSA. Considerable spectral-luminescent changes accompany the transformation of ABTN into HBTN that allows monitoring conjugation reaction. In presence of DNA and BSA the HBTN increases its emission in 15 and 4 times respectively and becomes strongly fluorescent. The conditions for labeling are developed and a model conjugate of HBTN dye with BSA is synthesized. It was shown that using of HBTN dye as a fluorescent label allows detection by eye of about 3 mug/band of BSA on polyacrylamide gel upon UV-irradiation.

  13. Robust method for 3D arterial spin labeling in mice.

    PubMed

    Chugh, Brige Paul; Bishop, Jonathan; Zhou, Yu-Qing; Wu, Jian; Henkelman, R Mark; Sled, John G

    2012-07-01

    Arterial spin labeling is a versatile perfusion quantification methodology, which has the potential to provide accurate characterization of cerebral blood flow (CBF) in mouse models. However, a paucity of physiological data needed for accurate modeling, more stringent requirements for gradient performance, and strong artifacts introduced by magnetization transfer present special challenges for accurate CBF mapping in the mouse. This article describes robust mapping of CBF over three-dimensional brain regions using amplitude-modulated continuous arterial spin labeling. To provide physiological data for CBF modeling, the carotid artery blood velocity distribution was characterized using pulsed-wave Doppler ultrasound. These blood velocity measurements were used in simulations that optimize inversion efficiency for parameters meeting MRI gradient duty cycle constraints. A rapid slice positioning algorithm was developed and evaluated to provide accurate positioning of the labeling plane. To account for enhancement of T(1) due to magnetization transfer, a binary spin bath model of magnetization transfer was used to provide a more accurate estimate of CBF. Finally, a study of CBF was conducted on 10 mice with findings of highly reproducible inversion efficiency (mean ± standard-error-of-the-mean, 0.67 ± 0.03), statistically significant variation in CBF over 12 brain regions (P < 0.0001) and a mean ± standard-error-of-the-mean whole brain CBF of 219 ± 6 mL/100 g/min.

  14. Water-Soluble Lipophilic MR Contrast Agents for Cell Membrane Labeling

    PubMed Central

    Carney, Christiane E.; MacRenaris, Keith W.; Meade, Thomas J.

    2015-01-01

    Long-term cell tracking with MR imaging necessitates the development of contrast agents that both label and are retained by cells. One promising strategy for long-term cell labeling is the development of lipophilic Gd(III)-based contrast agents that anchor into the cell membrane. We have previously reported the efficacy of monomeric and multimeric lipophilic agents and showed that the monomeric agents have improved labeling and contrast enhancement of cell populations. Here, we report on the synthesis, characterization, and in vitro testing of a series of monomeric lipophilic contrast agents with varied alkyl chain compositions. We show that these agents disperse in water, localize to the cell membrane, and label HeLa and MCF7 cells effectively. Additionally, these agents have up to 10-fold improved retention in cells compared to clinically available ProHance®. PMID:26215869

  15. 16 CFR 303.15 - Required label and method of affixing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... CONGRESS RULES AND REGULATIONS UNDER THE TEXTILE FIBER PRODUCTS IDENTIFICATION ACT § 303.15 Required label and method of affixing. (a) A label is required to be affixed to each textile product and, where..., resale and until sold and delivered to the ultimate consumer. (b) Each textile fiber product with a...

  16. A New First-Scan Method for Two-Scan Labeling Algorithms

    NASA Astrophysics Data System (ADS)

    He, Lifeng; Chao, Yuyan; Suzuki, Kenji

    This paper proposes a new first-scan method for two-scan labeling algorithms. In the first scan, our proposed method first scans every fourth image line, and processes the scan line and its two neighbor lines. Then, it processes the remaining lines from top to bottom one by one. Our method decreases the average number of times that must be checked to process a foreground pixel will; thus, the efficiency of labeling can be improved.

  17. The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method.

    PubMed

    do Nascimento, Cássio; Santos Barbosa, Rodrigo Edson; Mardegan Issa, João Paulo; Watanabe, Evandro; Yoko Ito, Izabel; Monesi, Nadia; Albuquerque Junior, Rubens Ferreira de

    2008-01-01

    Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique.

  18. LC/MS Method for the Determination of Stable Isotope Labeled Promethazine in Human Plasma

    NASA Technical Reports Server (NTRS)

    Zuwei, Wang; Boyd, Jason; Berens, Kurt L.; Putcha, Lakshmi

    2004-01-01

    Promethazine (PMZ) is taken by astronauts orally (PO), intramuscularly (IM) or rectally (PR) for space motion sickness. LC/MS method was developed with off-line solid phase extraction to measure plasma concentrations of PMZ given as stable isotope-labeled (SIL) formulations by the three different routes of administration simultaneously. Samples (0.5ml) were loaded on to Waters Oasis HLB co-polymer cartridges and eluted with 1.0 mL methanol. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 6 min. Acetonitrile/ ammonium acetate (30 mM) in water (3:2, v/v), pH 5.6 plus or minus 0.1, was used as the mobile phase for separation. Concentrations of PMZ, PMZ-d4 and PMZ-d7 and chlorpromazine (internal standard) were determined using a Micromass ZMD single quadrupole mass spectrometer with Electrospray Ionization (ESI). ESI mass spectra were acquired in positive ion mode with selected ion monitoring of [M+ H]dot plus. The method is rapid, reproducible and the assay specific parameters are listed in a table. A novel, sensitive and specific method for the measurement of PMZ and SIL PMZ in human plasma is reported.

  19. Use of an automated chromium reduction system for hydrogen isotope ratio analysis of physiological fluids applied to doubly labeled water analysis.

    PubMed

    Schoeller, D A; Colligan, A S; Shriver, T; Avak, H; Bartok-Olson, C

    2000-09-01

    The doubly labeled water method is commonly used to measure total energy expenditure in free-living subjects. The method, however, requires accurate and precise deuterium abundance determinations, which can be laborious. The aim of this study was to evaluate a fully automated, high-throughput, chromium reduction technique for the measurement of deuterium abundances in physiological fluids. The chromium technique was compared with an off-line zinc bomb reduction technique and also subjected to test-retest analysis. Analysis of international water standards demonstrated that the chromium technique was accurate and had a within-day precision of <1 per thousand. Addition of organic matter to water samples demonstrated that the technique was sensitive to interference at levels between 2 and 5 g l(-1). Physiological samples could be analyzed without this interference, plasma by 10000 Da exclusion filtration, saliva by sedimentation and urine by decolorizing with carbon black. Chromium reduction of urine specimens from doubly labeled water studies indicated no bias relative to zinc reduction with a mean difference in calculated energy expenditure of -0.2 +/- 3.9%. Blinded reanalysis of urine specimens from a second doubly labeled water study demonstrated a test-retest coefficient of variation of 4%. The chromium reduction method was found to be a rapid, accurate and precise method for the analysis of urine specimens from doubly labeled water. Copyright 2000 John Wiley & Sons, Ltd.

  20. Using power spectrum analysis to evaluate (18)O-water labeling data acquired from low resolution mass spectrometers.

    PubMed

    Sadygov, Rovshan G; Zhao, Yingxin; Haidacher, Sigmund J; Starkey, Jonathan M; Tilton, Ronald G; Denner, Larry

    2010-08-06

    We describe a method for ratio estimations in (18)O-water labeling experiments acquired from low resolution isotopically resolved data. The method is implemented in a software package specifically designed for use in experiments making use of zoom-scan mode data acquisition. Zoom-scan mode data allow commonly used ion trap mass spectrometers to attain isotopic resolution, which makes them amenable to use in labeling schemes such as (18)O-water labeling, but algorithms and software developed for high resolution instruments may not be appropriate for the lower resolution data acquired in zoom-scan mode. The use of power spectrum analysis is proposed as a general approach that may be uniquely suited to these data types. The software implementation uses a power spectrum to remove high-frequency noise and band-filter contributions from coeluting species of differing charge states. From the elemental composition of a peptide sequence, we generate theoretical isotope envelopes of heavy-light peptide pairs in five different ratios; these theoretical envelopes are correlated with the filtered experimental zoom scans. To automate peptide quantification in high-throughput experiments, we have implemented our approach in a computer program, MassXplorer. We demonstrate the application of MassXplorer to two model mixtures of known proteins and to a complex mixture of mouse kidney cortical extract. Comparison with another algorithm for ratio estimations demonstrates the increased precision and automation of MassXplorer.

  1. Using Power Spectrum Analysis to Evaluate 18O-Water Labeling Data Acquired from Low Resolution Mass Spectrometers

    PubMed Central

    Sadygov, Rovshan G.; Zhao, Yingxin; Haidacher, Sigmund J.; Starkey, Jonathan M.; Tilton, Ronald G.; Denner, Larry

    2010-01-01

    We describe a method for ratio estimations in 18O-water labeling experiments acquired from low resolution isotopically resolved data. The method is implemented in a software package specifically designed for use in experiments making use of zoom-scan mode data acquisition. Zoom-scan mode data allows commonly used ion trap mass spectrometers to attain isotopic resolution, which make them amenable to use in labeling schemes such as 18O-water labeling, but algorithms and software developed for high resolution instruments may not be appropriate for the lower resolution data acquired in zoom-scan mode. The use of power spectrum analysis is proposed as a general approach which may be uniquely suited to these data types. The software implementation uses power spectrum to remove high-frequency noise, and band-filter contributions from co-eluting species of differing charge states. From the elemental composition of a peptide sequence we generate theoretical isotope envelopes of heavy-light peptide pairs in five different ratios; these theoretical envelopes are correlated with the filtered experimental zoom scans. To automate peptide quantification in high-throughput experiments, we have implemented our approach in a computer program, MassXplorer. We demonstrate the application of MassXplorer to two model mixtures of known proteins, and to a complex mixture of mouse kidney cortical extract. Comparison with another algorithm for ratio estimations demonstrates the increased precision and automation of MassXplorer. PMID:20568695

  2. A method to label biological molecules with dsDNA coated gold nanoparticles.

    PubMed

    Xing, Ming; Li, Fangliang; Dong, Yingshan; Yu, Zhenxiang; Chen, Xia

    2015-03-01

    We described a new method to label biological molecules using gold nanoparticles (GNPs) and double stranded DNA. Researchers can conveniently label their own samples with GNPs using this method. The label is based on dsDNA with a 93.5% coverage of GNPs (dsDNA:GNP = 303:1). Antigens, streptavidin and biotin were labeled on GNPs and the success of the method was investigated with agarose gel electrophoresis, laser particle size analysis and ultraviolet spectrophotometry. These analyses confirmed that biological molecules were successfully bound to the GNPs. These molecules retained their biological activity and were able to detect targets on PVDF and NC membranes with excellent selectivity and low levels of background. Modified GNPs were also able to detect targets on nylon membranes, but with some degree of false positives. The maximum limit of detection was 25 ng proteins.

  3. Heavy water labeling of keratin as a non-invasive biomarker of skin turnover in vivo in rodents and humans.

    PubMed

    Lindwall, Glen; Hsieh, Elaine A; Misell, Lisa M; Chai, Christine M; Turner, Scott M; Hellerstein, Marc K

    2006-04-01

    Measurement of skin turnover has been problematic in humans. Heavy water (2H2O) labeling has recently been developed as a safe, simple method to study in vivo kinetics of many biosynthetic processes, including DNA and protein synthesis. Here, we apply this approach to the measurement of 2H incorporation into skin keratin and show close agreement between keratin and keratinocyte turnover data in the epidermis of rodents. Elevated turnover rates of both keratin and keratinocytes were observed in the epidermis of the flaky skin mouse, although topical treatments effective in human psoriasis had no effect on either turnover rate in these mice. In humans, keratin turnover was monitored non-invasively by serial tape stripping during and after 2H2O labeling. Kinetic data were consistent with previous estimates of epidermal turnover, with a lag time of 18 days before label appeared at the skin surface and a transit time of 4-5 weeks. Variability in skin keratin turnover rates was present among healthy individuals. In summary, 2H2O labeling of skin keratin represents a non-invasive approach for assessing skin turnover dynamics in pre-clinical models and in human subjects.

  4. Comparison of Test Procedures and Energy Efficiency Criteria in Selected International Standards and Labeling Programs for Clothes Washers, Water Dispensers, Vending Machines and CFLs

    SciTech Connect

    Fridley, David; Zheng, Nina; Zhou, Nan

    2010-06-01

    Since the late 1970s, energy labeling programs and mandatory energy performance standards have been used in many different countries to improve the efficiency levels of major residential and commercial equipment. As more countries and regions launch programs covering a greater range of products that are traded worldwide, greater attention has been given to harmonizing the specific efficiency criteria in these programs and the test methods for measurements. For example, an international compact fluorescent light (CFL) harmonization initiative was launched in 2006 to focus on collaboration between Australia, China, Europe and North America. Given the long history of standards and labeling programs, most major energy-consuming residential appliances and commercial equipment are already covered under minimum energy performance standards (MEPS) and/or energy labels. For these products, such as clothes washers and CFLs, harmonization may still be possible when national MEPS or labeling thresholds are revised. Greater opportunity for harmonization exists in newer energy-consuming products that are not commonly regulated but are under consideration for new standards and labeling programs. This may include commercial products such as water dispensers and vending machines, which are only covered by MEPS or energy labels in a few countries or regions. As China continues to expand its appliance standards and labeling programs and revise existing standards and labels, it is important to learn from recent international experiences with efficiency criteria and test procedures for the same products. Specifically, various types of standards and labeling programs already exist in North America, Europe and throughout Asia for products in China's 2010 standards and labeling programs, namely clothes washers, water dispensers, vending machines and CFLs. This report thus examines similarities and critical differences in energy efficiency values, test procedure specifications and other

  5. Energetics during hatchling dispersal of the olive ridley turtle Lepidochelys olivacea using doubly labeled water.

    PubMed

    Clusella Trullas, Susana; Spotila, James R; Paladino, Frank V

    2006-01-01

    Studies of metabolism are central to the understanding of the ecology, behavior, and evolution of reptiles. This study focuses on one phase of the sea turtle life cycle, hatchling dispersal, and gives insight into energetic constraints that dispersal imposes on hatchlings. Hatchling dispersal is an energetically expensive phase in the life cycle of the olive ridley turtle Lepidochelys olivacea. Field metabolic rates (FMRs), determined using the doubly labeled water (DLW) method, for L. olivacea hatchlings digging out of their nest chamber, crawling at the sand surface, and swimming were five, four, and seven times, respectively, the resting metabolic rate (RMR). The cost of swimming was 1.5 and 1.8 times the cost of the digging and crawling phases, respectively, and we estimated that if L. olivacea hatchlings swim at frenzy levels, they can rely on yolk reserves to supply energy for only 3-6 d once they reach the ocean. We compared our RMR and FMR values by establishing an interspecific RMR mass-scaling relationship for a wide range of species in the order Testudines and found a scaling exponent of 1.06. This study demonstrates the feasibility of using the DLW method to estimate energetic costs of free-living sea turtle hatchlings and emphasizes the need for metabolic studies in various life-history stages.

  6. A comparative study of fat storage quantitation in nematode Caenorhabditis elegans using label and label-free methods.

    PubMed

    Yen, Kelvin; Le, Thuc T; Bansal, Ankita; Narasimhan, Sri Devi; Cheng, Ji-Xin; Tissenbaum, Heidi A

    2010-09-16

    of fat stores, but rather the sequestration of dyes in lysosome-related organelles. In contrast, fixative staining methods provide reproducible data but are prone to errors due to the interference of autofluorescent species and the non-specific staining of cellular structures other than fat stores. Importantly, both growth conditions and developmental stage should be considered when comparing methods of C. elegans lipid storage. Taken together, we confirm that CARS microscopy provides a direct, non-invasive, and label-free means to quantitatively analyze fat storage in living C. elegans.

  7. A Comparative Study of Fat Storage Quantitation in Nematode Caenorhabditis elegans Using Label and Label-Free Methods

    PubMed Central

    Bansal, Ankita; Narasimhan, Sri Devi; Cheng, Ji-Xin; Tissenbaum, Heidi A.

    2010-01-01

    of fat stores, but rather the sequestration of dyes in lysosome-related organelles. In contrast, fixative staining methods provide reproducible data but are prone to errors due to the interference of autofluorescent species and the non-specific staining of cellular structures other than fat stores. Importantly, both growth conditions and developmental stage should be considered when comparing methods of C. elegans lipid storage. Taken together, we confirm that CARS microscopy provides a direct, non-invasive, and label-free means to quantitatively analyze fat storage in living C. elegans. PMID:20862331

  8. Validation of Web-Based Physical Activity Measurement Systems Using Doubly Labeled Water

    PubMed Central

    Yamaguchi, Yukio; Yamada, Yosuke; Tokushima, Satoru; Hatamoto, Yoichi; Sagayama, Hiroyuki; Kimura, Misaka; Higaki, Yasuki; Tanaka, Hiroaki

    2012-01-01

    Background Online or Web-based measurement systems have been proposed as convenient methods for collecting physical activity data. We developed two Web-based physical activity systems—the 24-hour Physical Activity Record Web (24hPAR WEB) and 7 days Recall Web (7daysRecall WEB). Objective To examine the validity of two Web-based physical activity measurement systems using the doubly labeled water (DLW) method. Methods We assessed the validity of the 24hPAR WEB and 7daysRecall WEB in 20 individuals, aged 25 to 61 years. The order of email distribution and subsequent completion of the two Web-based measurements systems was randomized. Each measurement tool was used for a week. The participants’ activity energy expenditure (AEE) and total energy expenditure (TEE) were assessed over each week using the DLW method and compared with the respective energy expenditures estimated using the Web-based systems. Results The mean AEE was 3.90 (SD 1.43) MJ estimated using the 24hPAR WEB and 3.67 (SD 1.48) MJ measured by the DLW method. The Pearson correlation for AEE between the two methods was r = .679 (P < .001). The Bland-Altman 95% limits of agreement ranged from –2.10 to 2.57 MJ between the two methods. The Pearson correlation for TEE between the two methods was r = .874 (P < .001). The mean AEE was 4.29 (SD 1.94) MJ using the 7daysRecall WEB and 3.80 (SD 1.36) MJ by the DLW method. The Pearson correlation for AEE between the two methods was r = .144 (P = .54). The Bland-Altman 95% limits of agreement ranged from –3.83 to 4.81 MJ between the two methods. The Pearson correlation for TEE between the two methods was r = .590 (P = .006). The average input times using terminal devices were 8 minutes and 10 seconds for the 24hPAR WEB and 6 minutes and 38 seconds for the 7daysRecall WEB. Conclusions Both Web-based systems were found to be effective methods for collecting physical activity data and are appropriate for use in epidemiological studies. Because the measurement

  9. Active-Q: Validation of the Web-Based Physical Activity Questionnaire Using Doubly Labeled Water

    PubMed Central

    Trolle Lagerros, Ylva; Christensen, Sara Elisabeth; Möller, Elisabeth; Wright, Antony; Sjölander, Arvid; Bälter, Katarina

    2012-01-01

    Background Increased use of the Internet provides new opportunities for collecting data in large studies. The aim of our new Web-based questionnaire, Active-Q, is to assess total physical activity and inactivity in adults. Active-Q assesses habitual activity during the past year via questions in four different domains: (1) daily occupation, (2) transportation to and from daily occupation, (3) leisure time activities, and (4) sporting activities. Objective The objective of our study is to validate Active-Q’s energy expenditure estimates using the doubly labeled water (DLW) method, and to assess the reproducibility of Active-Q by comparing the results of the questionnaire completed by the same group on two occasions. Methods The validity and reproducibility of Active-Q were assessed in a group of 37 individuals, aged 20 to 65 years. Active-Q was distributed via email to the participants. The total energy expenditure of the participants was assessed using DLW for 11 consecutive days. Results The median time to complete Active-Q was 6.1 minutes. The majority of participants (27/37, 73%) reported that the questionnaire was “easy” or “very easy” to answer. On average, Active-Q overestimated the total daily energy expenditure by 440 kJ compared with the DLW. The Spearman correlation between the two methods was r = 0.52 (P < .001). The intraclass correlation coefficient for total energy expenditure between the results of Active-Q completed on two occasions was 0.83 (95% CI 0.73-0.93). Conclusions Active-Q is a valid and reproducible method of assessing total energy expenditure. It is also a user-friendly method and suitable for Web-based data collection in large epidemiological studies. PMID:22356755

  10. A simple fluorescence labeling method for studies of protein oxidation, protein modification, and proteolysis.

    PubMed

    Pickering, Andrew M; Davies, Kelvin J A

    2012-01-15

    Proteins are sensitive to oxidation, and oxidized proteins are excellent substrates for degradation by proteolytic enzymes such as the proteasome and the mitochondrial Lon protease. Protein labeling is required for studies of protein turnover. Unfortunately, most labeling techniques involve (3)H or (14)C methylation, which is expensive, exposes researchers to radioactivity, generates large amounts of radioactive waste, and allows only single-point assays because samples require acid precipitation. Alternative labeling methods have largely proven unsuitable, either because the probe itself is modified by the oxidant(s) being studied or because the alternative labeling techniques are too complex or too costly for routine use. What is needed is a simple, quick, and cheap labeling technique that uses a non-radioactive marker, binds strongly to proteins, is resistant to oxidative modification, and emits a strong signal. We have devised a new reductive method for labeling free carboxyl groups of proteins with the small fluorophore 7-amino-4-methycoumarin (AMC). When bound to target proteins, AMC fluoresces very weakly but when AMC is released by proteinases, proteases, or peptidases, it fluoresces strongly. Thus, without acid precipitation, the proteolysis of any target protein can be studied continuously, in multiwell plates. In direct comparisons, (3)H-labeled proteins and AMC-labeled proteins exhibited essentially identical degradation patterns during incubation with trypsin, cell extracts, and purified proteasome. AMC-labeled proteins are well suited to studying increased proteolytic susceptibility after protein modification, because the AMC-protein bond is resistant to oxidizing agents such as hydrogen peroxide and peroxynitrite and is stable over time and to extremes of pH, temperature (even boiling), freeze-thaw, mercaptoethanol, and methanol. Copyright © 2011. Published by Elsevier Inc.

  11. Method for rapid base sequencing in DNA and RNA with two base labeling

    DOEpatents

    Jett, James H.; Keller, Richard A.; Martin, John C.; Posner, Richard G.; Marrone, Babetta L.; Hammond, Mark L.; Simpson, Daniel J.

    1995-01-01

    Method for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used to replicate a single DNA or RNA strand to be sequenced. The bases are then sequentially cleaved from the replicated strand, excited with a chosen spectrum of electromagnetic radiation, and the fluorescence from individual, tagged bases detected in the order of cleavage from the strand.

  12. Method for rapid base sequencing in DNA and RNA with two base labeling

    DOEpatents

    Jett, J.H.; Keller, R.A.; Martin, J.C.; Posner, R.G.; Marrone, B.L.; Hammond, M.L.; Simpson, D.J.

    1995-04-11

    A method is described for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used to replicate a single DNA or RNA strand to be sequenced. The bases are then sequentially cleaved from the replicated strand, excited with a chosen spectrum of electromagnetic radiation, and the fluorescence from individual, tagged bases detected in the order of cleavage from the strand. 4 figures.

  13. Validation of Web-based physical activity measurement systems using doubly labeled water.

    PubMed

    Namba, Hideyuki; Yamaguchi, Yukio; Yamada, Yosuke; Tokushima, Satoru; Hatamoto, Yoichi; Sagayama, Hiroyuki; Kimura, Misaka; Higaki, Yasuki; Tanaka, Hiroaki

    2012-09-25

    Online or Web-based measurement systems have been proposed as convenient methods for collecting physical activity data. We developed two Web-based physical activity systems-the 24-hour Physical Activity Record Web (24hPAR WEB) and 7 days Recall Web (7daysRecall WEB). To examine the validity of two Web-based physical activity measurement systems using the doubly labeled water (DLW) method. We assessed the validity of the 24hPAR WEB and 7daysRecall WEB in 20 individuals, aged 25 to 61 years. The order of email distribution and subsequent completion of the two Web-based measurements systems was randomized. Each measurement tool was used for a week. The participants' activity energy expenditure (AEE) and total energy expenditure (TEE) were assessed over each week using the DLW method and compared with the respective energy expenditures estimated using the Web-based systems. The mean AEE was 3.90 (SD 1.43) MJ estimated using the 24hPAR WEB and 3.67 (SD 1.48) MJ measured by the DLW method. The Pearson correlation for AEE between the two methods was r = .679 (P < .001). The Bland-Altman 95% limits of agreement ranged from -2.10 to 2.57 MJ between the two methods. The Pearson correlation for TEE between the two methods was r = .874 (P < .001). The mean AEE was 4.29 (SD 1.94) MJ using the 7daysRecall WEB and 3.80 (SD 1.36) MJ by the DLW method. The Pearson correlation for AEE between the two methods was r = .144 (P = .54). The Bland-Altman 95% limits of agreement ranged from -3.83 to 4.81 MJ between the two methods. The Pearson correlation for TEE between the two methods was r = .590 (P = .006). The average input times using terminal devices were 8 minutes and 10 seconds for the 24hPAR WEB and 6 minutes and 38 seconds for the 7daysRecall WEB. Both Web-based systems were found to be effective methods for collecting physical activity data and are appropriate for use in epidemiological studies. Because the measurement accuracy of the 24hPAR WEB was moderate to high, it could

  14. Validation of laboratory-scale recycling test method of paper PSA label products

    Treesearch

    Carl Houtman; Karen Scallon; Richard Oldack

    2008-01-01

    Starting with test methods and a specification developed by the U.S. Postal Service (USPS) Environmentally Benign Pressure Sensitive Adhesive Postage Stamp Program, a laboratory-scale test method and a specification were developed and validated for pressure-sensitive adhesive labels, By comparing results from this new test method and pilot-scale tests, which have been...

  15. Energy Expenditure in Older Adults Who Are Frail: A Doubly Labeled Water Study.

    PubMed

    de Carvalho Bastone, Alessandra; Ferriolli, Eduardo; Pfrimer, Karina; de Souza Moreira, Bruno; Diz, Juliano Bergamaschine Mata; Dias, João Marcos Domingues; Dias, Rosângela Corrêa

    2017-08-03

    Frailty is a common and important geriatric syndrome, distinct from any single chronic disease, and an independent predictor of mortality. It is characterized by age-associated decline in physiological reserve and function across multiple systems, culminating in a vicious cycle of altered energy expenditure. The total energy expenditure (TEE) of an individual includes the resting metabolic rate (RMR), the thermic effect of feeding, and the energy expenditure in physical activity (PAEE). The investigation of the energy expenditure of older adults who are frail is essential for better understanding the syndrome. Therefore, we compared the RMR, the PAEE, the physical activity level (PAL), and the TEE of older adults who were frail with those who were not frail. A cross-sectional study was conducted with 26 community-dwelling older adults (66-86 years of age). Older adults in the frail and nonfrail groups were matched for age and gender, and the matched pairs were randomly selected to continue the study. The RMR was measured by indirect calorimetry. The TEE was obtained by the multipoint, doubly labeled water method. After collecting a baseline urine sample, each participant received an oral dose of doubly labeled water composed of deuterium oxide and oxygen-18 (H2O). Subsequently, urine samples were collected on the 1st, 2nd, 3rd, 7th, 12th, 13th, and 14th days after the baseline collection and analyzed by mass spectrometry. The older adults who were frail presented significantly lower PAEE (1453.7 [1561.9] vs 3336.1 [1829.3] kj/d, P < .01), PAL (1.4 [0.3] vs 1.9 [0.6], P = .04), and TEE (7919.0 [2151.9] vs 10442.4 [2148.0] kj/d, P < .01) than the older adults who were nonfrail. There was no difference in their RMRs (5673.3 [1569.2] vs 6062.0 [1891.7] kj/d, P = .57). Frailty has been associated with a smaller lean body mass and with a disease-related hypermetabolic state, which might explain the lack of difference in the RMR. The PAL of the older adults who were frail

  16. Systematic review: reliability of compendia methods for off-label oncology indications.

    PubMed

    Abernethy, Amy P; Raman, Gowri; Balk, Ethan M; Hammond, Julia M; Orlando, Lori A; Wheeler, Jane L; Lau, Joseph; McCrory, Douglas C

    2009-03-03

    The Centers for Medicare & Medicaid Services limit coverage of cancer drugs for off-label indications to indications listed in specified compendia. To assess whether compendia provide comprehensive, research-based, and timely information for off-label prescribing in oncology. 6 drug compendia, English-language literature searches of MEDLINE and the Cochrane Central Register of Controlled Trials from 2006 and 2008, and American Society of Clinical Oncology annual meeting abstracts from 2004 to 2007. Data Assessment: The compendia's stated methods, literature related to off-label indications of 14 cancer drugs in 2006, updated literature related to 1 off-label indication between 2006 and 2008, and completeness of compendia content and citations were assessed. The compendia's stated methods varied greatly from their actual practices. Compendia cited little of the available evidence, often neither the most recent nor that of highest methodological quality. Compendia differed in evidence cited, terminology, detail, presentation, and referencing. For the 14 off-label indications studied, the compendia differed in the indications included and whether and how they recommended particular agents for particular types of cancer. Update schedules varied, and documentation practices made it difficult to determine whether and when compendia content was updated. For 1 indication, compendia citations did not increase between 2006 and 2008 despite newly published articles. The 2006 analysis was limited to 14 off-label indications; the 2008 update examined 1 indication. Only off-label indications for cancer drugs were included, and results cannot be generalized to noncancer drugs or indications. Oncologists rely on compendia for up-to-date access to evidence and reimbursement information for off-label indications. Current compendia lack transparency, cite little current evidence, and lack systematic methods to review or update evidence.

  17. Total energy expenditure as measured by doubly-labeled water in outpatients with bulimia nervosa.

    PubMed

    Kotler, L A; Devlin, M J; Matthews, D E; Walsh, B T

    2001-05-01

    This study measured total energy expenditure (TEE) in symptomatic outpatient women with bulimia nervosa and normal controls. The study aimed to test the conceptual model of bulimia nervosa as an illness characterized by a physiological state of starvation, despite normal weight. Total fat and fat-free mass were measured using hydrodensitometry and total energy expenditure was assessed via the doubly-labeled water method, in nine normal weight outpatient females with DSM-III-R bulimia nervosa and ten healthy female controls. Patients and controls were similar in age, body mass index, weight, lean body mass, and levels of exercise and general activity. Patients had an average baseline binge frequency of 14.7 episodes per week and purge frequency of 16.8 times per week, and had been ill for an average of 11.9 years. Group mean TEE did not differ between patients and controls (patients 2380 +/- 482 kcal/day, controls 2368 +/- 515 kcal day). Observed TEE in the bulimic subjects did not differ significantly from TEE predicted on the basis of data from the controls. This finding of normal TEE in symptomatic outpatients with bulimia nervosa is consistent with a previous study that found no difference in TEE in a sample of symptomatic inpatients with bulimia nervosa. These data suggest that the energy conserving metabolic adaptations characteristic of semi-starvation do not occur in patients with bulimia nervosa. Copyright 2001 by John Wiley & Sons, Inc.

  18. Physical activity, food intake, and body weight regulation: insights from doubly labeled water studies.

    PubMed

    Westerterp, Klaas R

    2010-03-01

    Body weight and energy balance can be maintained by adapting energy intake to changes in energy expenditure and vice versa, whereas short-term changes in energy expenditure are mainly caused by physical activity. This review investigates whether physical activity is affected by over- and undereating, whether intake is affected by an increase or a decrease in physical activity, and whether being overweight affects physical activity. The available evidence is based largely on studies that quantified physical activity with doubly labeled water. Overeating does not affect physical activity, while undereating decreases habitual or voluntary physical activity. Thus, it is easier to gain weight than to lose weight. An exercise-induced increase in energy requirement is typically compensated by increased energy intake, while a change to a more sedentary routine does not induce an equivalent reduction of intake and generally results in weight gain. Overweight and obese subjects tend to have similar activity energy expenditures to lean people despite being more sedentary. There are two ways in which the general population trend towards increasing body weight can be reversed: reduce intake or increase physical activity. The results of the present literature review indicate that eating less is the most effective method for preventing weight gain, despite the potential for a negative effect on physical activity when a negative energy balance is reached.

  19. Using undiagnosed data to enhance computerized breast cancer analysis with a three stage data labeling method

    NASA Astrophysics Data System (ADS)

    Sun, Wenqing; Tseng, Tzu-Liang; Zheng, Bin; Lure, Flemin; Wu, Teresa; Francia, Giulio; Cabrera, Sergio; Zhang, Jianying; Vélez-Reyesv, Miguel; Qian, Wei

    2014-03-01

    A novel three stage Semi-Supervised Learning (SSL) approach is proposed for improving performance of computerized breast cancer analysis with undiagnosed data. These three stages include: (1) Instance selection, which is barely used in SSL or computerized cancer analysis systems, (2) Feature selection and (3) Newly designed `Divide Co-training' data labeling method. 379 suspicious early breast cancer area samples from 121 mammograms were used in our research. Our proposed `Divide Co-training' method is able to generate two classifiers through split original diagnosed dataset (labeled data), and label the undiagnosed data (unlabeled data) when they reached an agreement. The highest AUC (Area Under Curve, also called Az value) using labeled data only was 0.832 and it increased to 0.889 when undiagnosed data were included. The results indicate instance selection module could eliminate untypical data or noise data and enhance the following semi-supervised data labeling performance. Based on analyzing different data sizes, it can be observed that the AUC and accuracy go higher with the increase of either diagnosed data or undiagnosed data, and reach the best improvement (ΔAUC = 0.078, ΔAccuracy = 7.6%) with 40 of labeled data and 300 of unlabeled data.

  20. Special Considerations for Measuring Energy Expenditure with Doubly Labeled Water under Atypical Conditions

    PubMed Central

    Bhutani, Surabhi; Racine, Natalie; Shriver, Tim; Schoeller, Dale A

    2016-01-01

    The global increase in the prevalence of obesity has dramatically increased interest in understanding the factors that influence human total energy expenditure (TEE). This in turn has increased interest in the doubly labeled water (DLW) method, a technique for measurement of total energy expenditure in free-living humans. The increasing use of this method is attributed to its portability, objectivity, minimal invasiveness, high accuracy and good precision. Although a relatively standard protocol for the method has emerged, the new generation of users often is unfamiliar with rationale behind aspects of the protocol as well as the approaches to avoid or correct for in situations that are not covered by the standard protocol procedure. The primary uncommon situations like introduction of isotopically different diet and fluids with or without geographical relocation, seasonal and temperature variations, activity level of participants etc. during or prior to the DLW measurements can lead to shift in baseline abundance of 2H and 18O or tracer elimination, resulting in moderate to large errors in the measured TEE. These unique situations call for special modifications to the conventional protocol to minimize errors. The objective of the present review was to assemble a list of frequently asked questions and the issues they represent, and then examine the available literature to describe and explain the modifications to the standard DLW protocol to maintain the method’s accuracy. This discussion of DLW protocol modification can be an excellent resource for investigators who intend to use this measurement technique for interesting and uncommon study designs. PMID:26962472

  1. Detection of Cu2+ in Water Based on Histidine-Gold Labeled Multiwalled Carbon Nanotube Electrochemical Sensor

    PubMed Central

    Zhu, Rilong; Zhou, Gangqiang; Tang, Fengxia; Wang, Yeyao

    2017-01-01

    Based on the strong interaction between histidine and copper ions and the signal enhancement effect of gold-labeling carbon nanotubes, an electrochemical sensor is established and used to measure copper ions in river water. In this study the results show that the concentrations of copper ion have well linear relationship with the peak current in the range of 10−11–10−7 mol/L, and the limit of detection is 10−12 mol/L. When using this method to detect copper ions in the Xiangjiang River, the test results are consistent with the atomic absorption method. This study shows that the sensor is convenient to be used in daily monitoring of copper ions in river water. PMID:28408929

  2. Detection of Cu(2+) in Water Based on Histidine-Gold Labeled Multiwalled Carbon Nanotube Electrochemical Sensor.

    PubMed

    Zhu, Rilong; Zhou, Gangqiang; Tang, Fengxia; Tong, Chunyi; Wang, Yeyao; Wang, Jinsheng

    2017-01-01

    Based on the strong interaction between histidine and copper ions and the signal enhancement effect of gold-labeling carbon nanotubes, an electrochemical sensor is established and used to measure copper ions in river water. In this study the results show that the concentrations of copper ion have well linear relationship with the peak current in the range of 10(-11)-10(-7 )mol/L, and the limit of detection is 10(-12 )mol/L. When using this method to detect copper ions in the Xiangjiang River, the test results are consistent with the atomic absorption method. This study shows that the sensor is convenient to be used in daily monitoring of copper ions in river water.

  3. Pulse-labelling trees to study carbon allocation dynamics: a review of methods, current knowledge and future prospects.

    PubMed

    Epron, Daniel; Bahn, Michael; Derrien, Delphine; Lattanzi, Fernando Alfredo; Pumpanen, Jukka; Gessler, Arthur; Högberg, Peter; Maillard, Pascale; Dannoura, Masako; Gérant, Dominique; Buchmann, Nina

    2012-06-01

    Pulse-labelling of trees with stable or radioactive carbon (C) isotopes offers the unique opportunity to trace the fate of labelled CO(2) into the tree and its release to the soil and the atmosphere. Thus, pulse-labelling enables the quantification of C partitioning in forests and the assessment of the role of partitioning in tree growth, resource acquisition and C sequestration. However, this is associated with challenges as regards the choice of a tracer, the methods of tracing labelled C in tree and soil compartments and the quantitative analysis of C dynamics. Based on data from 47 studies, the rate of transfer differs between broadleaved and coniferous species and decreases as temperature and soil water content decrease. Labelled C is rapidly transferred belowground-within a few days or less-and this transfer is slowed down by drought. Half-lives of labelled C in phloem sap (transfer pool) and in mature leaves (source organs) are short, while those of sink organs (growing tissues, seasonal storage) are longer. (13)C measurements in respiratory efflux at high temporal resolution provide the best estimate of the mean residence times of C in respiratory substrate pools, and the best basis for compartmental modelling. Seasonal C dynamics and allocation patterns indicate that sink strength variations are important drivers for C fluxes. We propose a conceptual model for temperate and boreal trees, which considers the use of recently assimilated C versus stored C. We recommend best practices for designing and analysing pulse-labelling experiments, and identify several topics which we consider of prime importance for future research on C allocation in trees: (i) whole-tree C source-sink relations, (ii) C allocation to secondary metabolism, (iii) responses to environmental change, (iv) effects of seasonality versus phenology in and across biomes, and (v) carbon-nitrogen interactions. Substantial progress is expected from emerging technologies, but the largest challenge

  4. Cell labeling and tracking method without distorted signals by phagocytosis of macrophages.

    PubMed

    Kang, Sun-Woong; Lee, Sangmin; Na, Jin Hee; Yoon, Hwa In; Lee, Dong-Eun; Koo, Heebeom; Cho, Yong Woo; Kim, Sun Hwa; Jeong, Seo Young; Kwon, Ick Chan; Choi, Kuiwon; Kim, Kwangmeyung

    2014-01-01

    Cell labeling and tracking are important processes in understanding biologic mechanisms and the therapeutic effect of inoculated cells in vivo. Numerous attempts have been made to label and track inoculated cells in vivo; however, these methods have limitations as a result of their biological effects, including secondary phagocytosis of macrophages and genetic modification. Here, we investigated a new cell labeling and tracking strategy based on metabolic glycoengineering and bioorthogonal click chemistry. We first treated cells with tetra-acetylated N-azidoacetyl-D-mannosamine to generate unnatural sialic acids with azide groups on the surface of the target cells. The azide-labeled cells were then transplanted to mouse liver, and dibenzyl cyclooctyne-conjugated Cy5 (DBCO-Cy5) was intravenously injected into mice to chemically bind with the azide groups on the surface of the target cells in vivo for target cell visualization. Unnatural sialic acids with azide groups could be artificially induced on the surface of target cells by glycoengineering. We then tracked the azide groups on the surface of the cells by DBCO-Cy5 in vivo using bioorthogonal click chemistry. Importantly, labeling efficacy was enhanced and false signals by phagocytosis of macrophages were reduced. This strategy will be highly useful for cell labeling and tracking.

  5. Cell Labeling and Tracking Method without Distorted Signals by Phagocytosis of Macrophages

    PubMed Central

    Kang, Sun-Woong; Lee, Sangmin; Na, Jin Hee; Yoon, Hwa In; Lee, Dong-Eun; Koo, Heebeom; Cho, Yong Woo; Kim, Sun Hwa; Jeong, Seo Young; Kwon, Ick Chan; Choi, Kuiwon; Kim, Kwangmeyung

    2014-01-01

    Cell labeling and tracking are important processes in understanding biologic mechanisms and the therapeutic effect of inoculated cells in vivo. Numerous attempts have been made to label and track inoculated cells in vivo; however, these methods have limitations as a result of their biological effects, including secondary phagocytosis of macrophages and genetic modification. Here, we investigated a new cell labeling and tracking strategy based on metabolic glycoengineering and bioorthogonal click chemistry. We first treated cells with tetra-acetylated N-azidoacetyl-D-mannosamine to generate unnatural sialic acids with azide groups on the surface of the target cells. The azide-labeled cells were then transplanted to mouse liver, and dibenzyl cyclooctyne-conjugated Cy5 (DBCO-Cy5) was intravenously injected into mice to chemically bind with the azide groups on the surface of the target cells in vivo for target cell visualization. Unnatural sialic acids with azide groups could be artificially induced on the surface of target cells by glycoengineering. We then tracked the azide groups on the surface of the cells by DBCO-Cy5 in vivo using bioorthogonal click chemistry. Importantly, labeling efficacy was enhanced and false signals by phagocytosis of macrophages were reduced. This strategy will be highly useful for cell labeling and tracking. PMID:24578725

  6. Label-free methods of reporting biomolecular interactions by optical biosensors.

    PubMed

    Citartan, Marimuthu; Gopinath, Subash C B; Tominaga, Junji; Tang, Thean-Hock

    2013-07-07

    Reporting biomolecular interactions has become part and parcel of many applications of science towards an in-depth understanding of disease and gene regulation. Apart from that, in diagnostic applications where biomolecules (antibodies and aptamers) are vastly applied, meticulous monitoring of biomolecular interaction is vital for clear-cut diagnosis. Several currently available methods of analyzing the interaction of the ligands with the appropriate analytes are aided by labeling using fluorescence or luminescence techniques. However, labeling is cumbersome and can occupy important binding sites of interactive molecules to be labeled, which may interfere with the conformational changes of the molecules and increase non-specificity. Optical-based sensing can provide an alternative way as a label-free procedure for monitoring biomolecular interactions. Optical sensors affiliated with different operating principles, including surface plasmon changes, scattering and interferometry, can impart a huge impact for in-house and point-of-care applications. This optical-based biosensing permits real-time monitoring, obviating the use of hazardous labeling molecules such as radioactive tags. Herein, label-free ways of reporting biomolecular interactions by various optical biosensors were gleaned.

  7. Energy expenditure estimated by accelerometry and doubly labeled water: do they agree?

    PubMed

    Leenders, Nicole Y; Sherman, William Michael; Nagaraja, Haikady N

    2006-12-01

    The purpose of this study was to compare energy expenditure derived from regression equations determined from accelerometry with energy expenditure obtained from the doubly labeled water method (DLW). Thirteen subjects participated in a 7-d protocol during which total daily energy expenditure (TDEE) was measured with DLW. Simultaneously, during the 7 d, subjects wore a Tritrac-R3D and an Actigraph (ACT). Pearson and concordance correlations and one-sample t-tests were used to determine the agreement of six Tritrac and eight ACT regression equations that convert body acceleration to energy expenditure with the DLW measurements. Tritrac TDEE determined from the different Tritrac regression equations under- and overestimated TDEE determined with DLW that ranged from -10 to +101%. For ACT, the percent difference between DLW and ACT-TDEE determined with the regression equation developed by Hendelman and Swartz were not statistically significantly different from zero. The mean of the difference was -2 and -4%, but the range of the difference was large for both equations, -29 to +24%. TDEE determined with the six other ACT equations were significantly different compared with DLW. Of the 14 different regression equations from the literature, only two developed for ACT compared favorably with DLW; however, the difference in TDEE between these two methods was variable and rather large. These results reemphasize the difficulty in converting body movement into energy expenditure on an individual basis from accelerometry. These results imply that researchers may want to avoid using accelerometers to predict energy expenditure in free-living conditions, instead using these instruments only to measure patterns of physical activity.

  8. DNA-labeled clay: A sensitive new method for tracing particle transport

    USGS Publications Warehouse

    Mahler, B.J.; Winkler, M.; Bennett, P.; Hillis, D.M.

    1998-01-01

    The behavior of mobile colloids and sediment in most natural environments remains poorly understood, in part because characteristics of existing sediment tracers limit their wide-spread use. Here we describe the development of a new approach that uses a DNA-labeled montmorillonite clay as a highly sensitive and selective sediment tracer that can potentially characterize sediment and colloid transport in a wide variety of environments, including marine, wetland, ground-water, and atmospheric systems. Characteristics of DNA in natural systems render it unsuitable as an aqueous tracer but admirably suited as a label for tracing particulates. The DNA-labeled-clay approach, using techniques developed from molecular biology, has extremely low detection limits, very specific detection, and a virtually infinite number of tracer signatures. Furthermore, DNA-labeled clay has the same physical characteristics as the particles it is designed to trace, it is environmentally benign, and it can be relatively inexpensively produced and detected. Our initial results show that short (500 base pair) strands of synthetically produced DNA reversibly adsorb to both Na-montmorillonite and powdered silica surfaces via a magnesium bridge. The DNA-montmorillonite surface complexes are stable in calcium-bicarbonate spring waters for periods of up to 18 days and only slowly desorb to the aqueous phase, whereas the silica surface complex is stable only in distilled water. Both materials readily release the adsorbed DNA in dilute EDTA solutions for amplification by the polymerase chain reaction (PCR) and quantification. The stability of the DNA-labeled clay complex suggests that this material would be appropriate for use as an extremely sensitive sediment tracer for flow periods of as long as 2 weeks, and possibly longer.

  9. Combustion method for assay of biological materials labeled with carbon-14 or tritium, or double-labeled

    NASA Technical Reports Server (NTRS)

    Huebner, L. G.; Kisieleski, W. E.

    1969-01-01

    Dry catalytic combustion at high temperatures is used for assaying biological materials labeled carbon-14 and tritium, or double-labeled. A modified oxygen-flask technique is combined with standard vacuum-line techniques and includes convenience of direct in-vial collection of final combustion products, giving quantitative recovery of tritium and carbon-14.

  10. A novel single-labeled fluorescent oligonucleotide probe for silver(I) ion detection in water, drugs, and food.

    PubMed

    Bian, Liujiao; Ji, Xu; Hu, Wei

    2014-05-28

    Due to the high toxicity of silver(I) ions, a method for the rapid, sensitive, and selective detection for silver(I) ions in water, pharmaceutical products, and food is of great importance. Herein, a novel single-labeled fluorescent oligonucleotide (OND) probe based on cytosine-Ag(I)-cytosine coordination and the inherent fluorescence quenching ability of the G-quadruplex is designed to detect silver(I) ions. The formation of a hairpin structure in the OND-Ag(I) complex brings the hexachloro fluorescein (HEX) labeled at the 5'-end of the OND probe close to the G-quadruplex located at the 3'-end of the OND probe, leading to a fluorescence quenching due to photoinduced electron transfer between HEX and the G-quadruplex. Through this method, silver(I) ions can be detected quantitatively, the linear response range is from 1 to 100 nmol/L with a detection limit of 50 pmol/L, and no obvious interference occurs with other metal ions with a 10-fold concentration. This assay is simple, sensitive, and selective, and it can be used to detect silver(I) ions in actual water, drug, and food samples.

  11. Comparison of cerebral blood flow measurement with [15O]-water positron emission tomography and arterial spin labeling magnetic resonance imaging: A systematic review.

    PubMed

    Fan, Audrey P; Jahanian, Hesamoddin; Holdsworth, Samantha J; Zaharchuk, Greg

    2016-05-01

    Noninvasive imaging of cerebral blood flow provides critical information to understand normal brain physiology as well as to identify and manage patients with neurological disorders. To date, the reference standard for cerebral blood flow measurements is considered to be positron emission tomography using injection of the [(15)O]-water radiotracer. Although [(15)O]-water has been used to study brain perfusion under normal and pathological conditions, it is not widely used in clinical settings due to the need for an on-site cyclotron, the invasive nature of arterial blood sampling, and experimental complexity. As an alternative, arterial spin labeling is a promising magnetic resonance imaging technique that magnetically labels arterial blood as it flows into the brain to map cerebral blood flow. As arterial spin labeling becomes more widely adopted in research and clinical settings, efforts have sought to standardize the method and validate its cerebral blood flow values against positron emission tomography-based cerebral blood flow measurements. The purpose of this work is to critically review studies that performed both [(15)O]-water positron emission tomography and arterial spin labeling to measure brain perfusion, with the aim of better understanding the accuracy and reproducibility of arterial spin labeling relative to the positron emission tomography reference standard.

  12. Comparison of electron paramagnetic resonance methods to determine distances between spin labels on human carbonic anhydrase II.

    PubMed Central

    Persson, M; Harbridge, J R; Hammarström, P; Mitri, R; Mårtensson, L G; Carlsson, U; Eaton, G R; Eaton, S S

    2001-01-01

    Four doubly spin-labeled variants of human carbonic anhydrase II and corresponding singly labeled variants were prepared by site-directed spin labeling. The distances between the spin labels were obtained from continuous-wave electron paramagnetic resonance spectra by analysis of the relative intensity of the half-field transition, Fourier deconvolution of line-shape broadening, and computer simulation of line-shape changes. Distances also were determined by four-pulse double electron-electron resonance. For each variant, at least two methods were applicable and reasonable agreement between methods was obtained. Distances ranged from 7 to 24 A. The doubly spin-labeled samples contained some singly labeled protein due to incomplete labeling. The sensitivity of each of the distance determination methods to the non-interacting component was compared. PMID:11371461

  13. An Efficient Site-Specific Method for Irreversible Covalent Labeling of Proteins with a Fluorophore.

    PubMed

    Liu, Jiaquan; Hanne, Jeungphill; Britton, Brooke M; Shoffner, Matthew; Albers, Aaron E; Bennett, Jared; Zatezalo, Rachel; Barfield, Robyn; Rabuka, David; Lee, Jong-Bong; Fishel, Richard

    2015-11-19

    Fluorophore labeling of proteins while preserving native functions is essential for bulk Förster resonance energy transfer (FRET) interaction and single molecule imaging analysis. Here we describe a versatile, efficient, specific, irreversible, gentle and low-cost method for labeling proteins with fluorophores that appears substantially more robust than a similar but chemically distinct procedure. The method employs the controlled enzymatic conversion of a central Cys to a reactive formylglycine (fGly) aldehyde within a six amino acid Formylglycine Generating Enzyme (FGE) recognition sequence in vitro. The fluorophore is then irreversibly linked to the fGly residue using a Hydrazinyl-Iso-Pictet-Spengler (HIPS) ligation reaction. We demonstrate the robust large-scale fluorophore labeling and purification of E.coli (Ec) mismatch repair (MMR) components. Fluorophore labeling did not alter the native functions of these MMR proteins in vitro or in singulo. Because the FGE recognition sequence is easily portable, FGE-HIPS fluorophore-labeling may be easily extended to other proteins.

  14. A novel facile method of labeling octreotide with 18F-fluorine

    PubMed Central

    Laverman, Peter; McBride, William J; Sharkey, Robert M; Eek, Annemarie; Joosten, Lieke; Oyen, Wim JG; Goldenberg, David M; Boerman, Otto C

    2010-01-01

    Several methods have been developed to label peptides with fluorine-18. However, in general these are laborious and require a multistep synthesis. We present a facile method based on the chelation of [18F]aluminum fluoride (“Al18F”) by NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid). The method is characterized by labeling NOTA-octreotide (IMP466) with 18F. Methods Octreotide was conjugated with the NOTA chelate and was labeled with 18F in a two-step, one-pot method. The labeling procedure was optimized with regard to the labeling buffer, peptide, and aluminum concentration. Radiochemical yield, specific activity, in vitro stability, and receptor affinity were determined. Biodistribution of 18F-IMP466 was studied in AR42J tumor-bearing mice and compared to that of 68Ga-labeled IMP466. In addition, microPET/CT images were acquired. Results IMP466 was labeled with “Al18F” in a single step with 50% yield. The labeled product was purified by HPLC to remove unbound “Al18F” and unlabeled peptide. The radiolabeling, including purification, was performed in 45 min. The specific activity was 45,000 GBq/mmol and the peptide was stable in serum for 4 h at 37° C. Labeling was performed at pH 4.1 in sodium citrate, sodium acetate, HEPES and MES buffer and was optimal in sodium acetate buffer. The apparent IC50 of the 19F-labeled IMP466 determined on AR42J cells was 3.6 nM. Biodistribution studies at 2 h p.i. showed a high tumor uptake of 18F-IMP466 (28.3 ± 5.2 %ID/g, tumor-to-blood ratio: 300 ± 90), which could be blocked by an excess of unlabeled peptide (8.6 ± 0.7%ID/g), indicating that the accumulation in the tumor was receptor-mediated. Biodistribution of 68Ga-IMP466 was similar to that of 18F-IMP466. 18F-IMP466 was stable in vivo, since bone uptake was only 0.4 ± 0.2 %ID/g, whereas free “Al18F” accumulated rapidly in the bone (36.9 ± 5.0 %ID/g at 2 h p.i.). MicroPET/CT scans showed excellent tumor delineation and high preferential accumulation in

  15. Oligonucleotide labeling methods. 3. Direct labeling of oligonucleotides employing a novel, non-nucleosidic, 2-aminobutyl-1,3-propanediol backbone.

    PubMed Central

    Nelson, P S; Kent, M; Muthini, S

    1992-01-01

    Novel CE-phosphoramidite (7a-e) and CPG (8a, c, d, e) reagents have been prepared from a unique 2-aminobutyl-1,3-propanediol backbone. The reagents have been used to directly label oligonucleotides with fluorescein, acridine, and biotin via automated DNA synthesis. The versatile 2-aminobutyl-1,3-propanediol backbone allows for labeling at any position (5', internal, and 3') during solid phase oligonucleotide synthesis. Multiple labels can be achieved by repetitive coupling cycles. Furthermore, the 3-carbon atom internucleotide phosphate distance is retained when inserted internally. Using this method, individual oligonucleotides possessing two and three different reporter molecules have been prepared. PMID:1475185

  16. A Method to Constrain Genome-Scale Models with 13C Labeling Data

    PubMed Central

    García Martín, Héctor; Kumar, Vinay Satish; Weaver, Daniel; Ghosh, Amit; Chubukov, Victor; Mukhopadhyay, Aindrila; Arkin, Adam; Keasling, Jay D.

    2015-01-01

    Current limitations in quantitatively predicting biological behavior hinder our efforts to engineer biological systems to produce biofuels and other desired chemicals. Here, we present a new method for calculating metabolic fluxes, key targets in metabolic engineering, that incorporates data from 13C labeling experiments and genome-scale models. The data from 13C labeling experiments provide strong flux constraints that eliminate the need to assume an evolutionary optimization principle such as the growth rate optimization assumption used in Flux Balance Analysis (FBA). This effective constraining is achieved by making the simple but biologically relevant assumption that flux flows from core to peripheral metabolism and does not flow back. The new method is significantly more robust than FBA with respect to errors in genome-scale model reconstruction. Furthermore, it can provide a comprehensive picture of metabolite balancing and predictions for unmeasured extracellular fluxes as constrained by 13C labeling data. A comparison shows that the results of this new method are similar to those found through 13C Metabolic Flux Analysis (13C MFA) for central carbon metabolism but, additionally, it provides flux estimates for peripheral metabolism. The extra validation gained by matching 48 relative labeling measurements is used to identify where and why several existing COnstraint Based Reconstruction and Analysis (COBRA) flux prediction algorithms fail. We demonstrate how to use this knowledge to refine these methods and improve their predictive capabilities. This method provides a reliable base upon which to improve the design of biological systems. PMID:26379153

  17. Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells

    PubMed Central

    Leiker, Merced; Suzuki, Gen; Iyer, Vijay S.; Canty, John M.; Lee, Techung

    2010-01-01

    Therapeutic implantation of mesenchymal stem cells (MSCs) is entering the realm of clinical trials for several human diseases, and yet much remains uncertain regarding their dynamic distribution and cell fate after in vivo application. Discrepancies in the literature can be attributed in part to the use of different cell labeling/tracking methods and cell administration protocols. To identify a stem cell detection method suitable for myocardial implantation in a large animal model, we experimented on three different MSC labeling methods: adenovirus-mediated expression of enhanced green fluorescence protein (EGFP) and β-galactosidase (LacZ), and nuclear staining with DAPI. Intramuscular and intracoronary administrations of labeled porcine MSCs identified the nuclear affinity dye to be a reliable stem cell tracking marker. Stem cell identification is facilitated by an optimized live cell labeling condition generating bright blue fluorescence sharply confined to the nucleus. DAPI-labeled MSCs retained full viability, ceased proliferation, and exhibited an increased differentiation potential. The labeled MSCs remained fully active in expressing key growth factor and cytokine genes, and notably exhibited enhanced expression of the chemokine receptor CXCR4 and its ligand SDF1, indicating their competency in response to tissue injury. Histological analysis revealed that approximately half a million MSCs or ∼2% of the administered MSCs remained localized in the normal pig heart 2 weeks after coronary infusion. That the vast majority of these identified MSCs were interstitial indicated the ability of MSCs to migrate across the coronary endothelium. No evidence was obtained indicating MSC differentiation to cardiomyocyte. PMID:19069634

  18. Method for treating waste water

    SciTech Connect

    Masaki, Y.; Odawara, Y.; Shimizu, N.

    1982-10-26

    The invention relates to an improvement of the floc-formation property of activated sludge contained in waste water. A waste water treatment process comprises steps culturing a novel strain-alcaligenes faecalis hrl-1-and adding the cultured cells to to-be-treated waste water.

  19. Energy requirements of lactating women derived from doubly labeled water and milk energy output.

    PubMed

    Butte, N F; Wong, W W; Hopkinson, J M

    2001-01-01

    Instead of using an incremental approach to assess the energy requirements of lactation, a more comprehensive approach may be taken by measuring total energy expenditure (TEE), milk energy output and energy mobilization from tissue stores. The latter approach avoids assumptions regarding energetic efficiency and changes in physical activity and adiposity. The purpose of this study was threefold: to assess the energy requirements of lactation; to compare these estimates with energy requirements in the nonpregnant, nonlactating state and to test for energetic adaptations in basal metabolic rate (BMR) and physical activity during the energy-demanding process of lactation. Milk production and composition, body weight and composition, TEE, BMR and physical activity levels were measured in 24 well-nourished women during exclusive breastfeeding at 3 mo postpartum and after the cessation of breastfeeding at 18 or 24 mo postpartum. TEE was measured by the doubly labeled water method, milk production by 3-d test-weighing, milk energy by bomb calorimetry on a 24-h milk sample, body composition by dual-energy x-ray absorptiometry and BMR by room respiration calorimetry. TEE, BMR and physical activity level (physical activity level = TEE/BMR) did not differ between the lactating and nonlactating state (TEE 10.0 +/- 1.5 versus 10.6 +/- 2.1 MJ/d). Mean milk energy output was equivalent to 2.02 +/- 0.33 MJ/d. Total energy requirements were greater during lactation than afterward (12.0 +/- 1.4 versus 10.6 +/- 2.1 MJ/d, P: = 0.002). Energy mobilization from tissue stores (-0.65 +/- 0.97 MJ/d) resulted in net energy requirements during lactation of 11.4 +/- 1.8 MJ/d. Because adaptations in basal metabolism and physical activity were not evident in these well-nourished women, energy requirements during lactation were met primarily from the diet and only partially by mobilization of tissue stores.

  20. New method for the selective labeling of erythrocytes in whole blood with Tc-99m

    DOEpatents

    Srivastava, S.C.; Babich, J.W.; Straub, R.; Richards, P.

    1984-01-27

    Method and kit are described for the preparation of /sup 99m/Tc labeled red blood cells using whole blood in a closed sterile system containing stannous tin in a form such that it will enter the red blood cells and be available therein for the reduction of technetium.

  1. 40 CFR 600.115-11 - Criteria for determining the fuel economy label calculation method.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... economy label calculation method. 600.115-11 Section 600.115-11 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) ENERGY POLICY FUEL ECONOMY AND GREENHOUSE GAS EXHAUST EMISSIONS OF MOTOR VEHICLES Fuel Economy and Carbon-Related Exhaust Emission Test Procedures § 600.115-11 Criteria...

  2. 40 CFR 600.115-11 - Criteria for determining the fuel economy label calculation method.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 30 2014-07-01 2014-07-01 false Criteria for determining the fuel economy label calculation method. 600.115-11 Section 600.115-11 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) ENERGY POLICY FUEL ECONOMY AND GREENHOUSE GAS EXHAUST EMISSIONS OF...

  3. A method for (13)C-labeling of metabolic carbohydrates within French bean leaves (Phaseolus vulgaris L.) for decomposition studies in soils.

    PubMed

    Girardin, Cyril; Rasse, Daniel P; Biron, Philippe; Ghashghaie, Jaleh; Chenu, Claire

    2009-06-01

    The molecular composition of plant residues is suspected to largely govern the fate of their constitutive carbon (C) in soils. Labile compounds, such as metabolic carbohydrates, are affected differently from recalcitrant and structural compounds by soil-C stabilisation mechanisms. Producing (13)C-enriched plant residues with specifically labeled fractions would help us to investigate the fate in soils of the constitutive C of these compounds. The objective of the present research was to test (13)C pulse chase labeling as a method for specifically enriching the metabolic carbohydrate components of plant residues, i.e. soluble sugars and starch. Bean plants were exposed to a (13)CO(2)-enriched atmosphere for 0.5, 1, 2, 3 and 21 h. The major soluble sugars were then determined on water-soluble extracts, and starch on HCl-hydrolysable extracts. The results show a quick differential labeling between water-soluble and water-insoluble compounds. For both groups, (13)C-labeling increased linearly with time. The difference in delta(13)C signature between water-soluble and insoluble fractions was 7 per thousand after 0.5 h and 70 per thousand after 21 h. However, this clear isotopic contrast masked a substantial labeling variability within each fraction. By contrast, metabolic carbohydrates on the one hand (i.e. soluble sugars + starch) and other fractions (essentially cell wall components) on the other hand displayed quite homogeneous signatures within fractions, and a significant difference in labeling between fractions: delta(13)C = 414 +/- 3.7 per thousand and 56 +/- 5.5 per thousand, respectively. Thus, the technique generates labeled plant residues displaying contrasting (13)C-isotopic signatures between metabolic carbohydrates and other compounds, with homogenous signatures within each group. Metabolic carbohydrates being labile compounds, our findings suggest that the technique is particularly appropriate for investigating the effect of compound lability on the long

  4. Estimating the impact of different cigarette package warning label policies: the auction method.

    PubMed

    Thrasher, James F; Rousu, Matthew C; Anaya-Ocampo, Rafael; Reynales-Shigematsu, Luz Myriam; Arillo-Santillán, Edna; Hernández-Avila, Mauricio

    2007-12-01

    The study estimated the reduction in demand associated with implementing cigarette package warning labels that contain imagery illustrating the consequences of smoking. The experimental auction method was used, wherein adult smokers in Mexico (n=89) placed separate bids on two packs of cigarettes: one with a text-only warning label and the other with a warning label that included text and a graphic image. Differences in the values attributed to each pack were assessed using t-tests and multivariate regression. The pack with the graphic image had a mean attributed value which was 17% lower ($3.21 pesos) than the pack with the text-only warning, and this difference remained statistically significant within subgroups defined by sociodemographics, amount of smoking, number of quit attempts, and levels of perceived smoking risks. In the multivariate model, the difference in attributed values was greater among females than males, but no such differences were found for other sociodemographic or smoking-related variables. The consistently lower value that smokers attributed to cigarette packages with the graphic warning label indicates that these labels are likely to reduce cigarette demand.

  5. A fast, low cost, and highly efficient fluorescent DNA labeling method using methyl green.

    PubMed

    Prieto, Daniel; Aparicio, Gonzalo; Morande, Pablo E; Zolessi, Flavio R

    2014-09-01

    The increasing need for multiple-labeling of cells and whole organisms for fluorescence microscopy has led to the development of hundreds of fluorophores that either directly recognize target molecules or organelles, or are attached to antibodies or other molecular probes. DNA labeling is essential to study nuclear-chromosomal structure, as well as for gel staining, but also as a usual counterstain in immunofluorescence, FISH or cytometry. However, there are currently few reliable red to far-red-emitting DNA stains that can be used. We describe herein an extremely simple, inexpensive and robust method for DNA labeling of cells and electrophoretic gels using the very well-known histological stain methyl green (MG). MG used in very low concentrations at physiological pH proved to have relatively narrow excitation and emission spectra, with peaks at 633 and 677 nm, respectively, and a very high resistance to photobleaching. It can be used in combination with other common DNA stains or antibodies without any visible interference or bleed-through. In electrophoretic gels, MG also labeled DNA in a similar way to ethidium bromide, but, as expected, it did not label RNA. Moreover, we show here that MG fluorescence can be used as a stain for direct measuring of viability by both microscopy and flow cytometry, with full correlation to ethidium bromide staining. MG is thus a very convenient alternative to currently used red-emitting DNA stains.

  6. Referential learning of French and Czech labels in African grey parrots (Psittacus erithacus): different methods yield contrasting results.

    PubMed

    Giret, Nicolas; Péron, Franck; Lindová, Jitka; Tichotová, Lenka; Nagle, Laurent; Kreutzer, Michel; Tymr, Frantisek; Bovet, Dalila

    2010-10-01

    Some African grey parrots (Psittacus erithacus), the most famous being Pepperberg's parrot Alex, are able to imitate human speech and produce labels referentially. In this study, the aim was to teach ten African grey parrots from two laboratories to label items. Training three parrots from the first laboratory for several months with the Model/Rival method, developed by Pepperberg, in which two humans interact in front of the subject to demonstrate the use of a label, led to disappointing results. Similarly, seven parrots from the second laboratory, having been trained with several variants of Model/Rival attained little success. After the informal observation of the efficiency of other methods (i.e. learning to imitate labels either spontaneously or with specific learning methods and use of these labels referentially), four different teaching methods were tested with two birds: the Model/Rival; Repetition/Association which consisted of repeating a label and presenting the item only when the parrot produced the label; Intuitive in which the experimenter handled an item and repeated its name in front of the subject; Diffusion in which labels with either variable or flat intonation were played back daily to parrots. One bird learned three labels, one of which was used referentially, with the Repetition/Association method. He learned one label non-referentially with the Model/Rival but no labels were acquired using the other methods. The second bird did not learn any labels. This study demonstrates that different methods can be efficient to teach labels referentially and it suggests that rearing conditions and interindividual variability are important features when assessing learning ability of African grey parrots.

  7. PRN 81-4: Label Improvement Program - Label Revisions to Accommodate New AOAC Methods of Chemical Analysis

    EPA Pesticide Factsheets

    This notice describes procedures EPA will use to ensure that pesticide label ingredient statements reflect percentages of ingredients determined by the most specific analytical techniques approved by the Association of Official Analytical Chemists (AOCA).

  8. A simple method of labeling amyloid β with quantum dots and ingestion of the labeled amyloid β by astrocytes.

    PubMed

    Zhang, Jing; Jia, Xing; Qing, Hong; Xie, Hai-Yan

    2013-01-25

    Steady labeling of amyloid beta (Aβ) is crucial for studying the ingestion and degradation of Aβ by astrocytes and unraveling a relevant regulation mechanism. Quantum dots (QDs) are an optimum labeling reagent for this because of their strong and steady fluorescence properties. In this paper, Aβ was labeled with QDs by a simple mixed incubation strategy, with a QD labeled Aβ complex (QDs-Aβ) being obtained. In the complex, QDs efficiently restrained the formation of β-folding and fibrils of Aβ, while the graininess, dispersivity and fluorescence properties of the QDs hardly changed. The fluorescence microscopy imaging results showed that the astrocytes could ingest the QDs-Aβ. The QDs and Aβ did not separate from each other during the ingestion process, and the Aβ could be degraded subsequently.

  9. Tracing cellulose elements adsorbed on composite cellulose biomaterials by a new labeling method.

    PubMed

    Joseleau, Jean-Paul; Chevalier-Billosta, Valérie; Ruel, Katia

    2008-03-01

    In view of tracing the fate of cellulose fine elements added to a suspension of cellulose fibers, a novel method for specific labeling of polysaccharides in a composite material was developed. The purpose was to visualize a given cellulose material within a cellulose mixture. The method consists of generating aldehyde groups in the chain by mild periodic acid oxidation followed by biotinylation of the carbonyls. Once added to the composite, the biotinylated molecules are labeled with streptavidin conjugated to a fluorescent probe for confocal microscopy, or streptavidin-gold for electron microscopy observations. In the present work, the fate of fresh fines (never-dried) and dead fines (dried) when they were added to a purified suspension of fibers was followed by observation of the labeling in confocal and electron microscopy. The differential mode of interaction of fresh fines and dead fines with the fibers was correlated to the mechanical characteristics measured on the corresponding papers. The versatility of the new labeling method and its possible generalization to other polysaccharides incorporated to a polysaccharide or nonpolysaccharide material should be of potential interest for the study of composite microstructure.

  10. A simple method for Alexa Fluor dye labelling of dengue virus.

    PubMed

    Zhang, Summer Li-Xin; Tan, Hwee-Cheng; Hanson, Brendon J; Ooi, Eng Eong

    2010-08-01

    Dengue virus causes frequent and cyclical epidemics throughout the tropics, resulting in significant morbidity and mortality rates. There is neither a specific antiviral treatment nor a vaccine to prevent epidemic transmission. The lack of a detailed understanding of the pathogenesis of the disease complicates these efforts. The development of methods to probe the interaction between the virus and host cells would thus be useful. Direct fluorescence labelling of virus would facilitate the visualization of the early events in virus-cell interaction. This report describes a simple method of labelling of dengue virus with Alexa Fluor succinimidyl ester dye dissolved directly in the sodium bicarbonate buffer that yielded highly viable virus after labelling. Alexa Fluor dyes have superior photostability and are less pH-sensitive than the common dyes, such as fluorescein and rhodamine, making them ideal for studies on cellular uptake and endosomal transport of the virus. The conjugation of Alexa Fluor dye did not affect the recognition of labelled dengue virus by virus-specific antibody and its putative receptors in host cells. This method could have useful applications in virological studies.

  11. Method and apparatus for tritiated water separation

    DOEpatents

    Nelson, D.A.; Duncan, J.B.; Jensen, G.A.

    1995-09-19

    The present invention is a membrane method and apparatus for separating isotopic water constituents from light water. The method involves providing a supported membrane of an aromatic polyphosphazene and pressurizing the water on one side of the membrane thereby forcing the light water through the supported membrane while isotopic water constituents are retained or vice versa. The apparatus of the present invention includes an aromatic polyphosphazene placed on a porous support and means for pressurizing water through the membrane while certain isotopic water constituents are retained. 1 fig.

  12. Method and apparatus for tritiated water separation

    DOEpatents

    Nelson, David A.; Duncan, James B.; Jensen, George A.

    1995-01-01

    The present invention is a membrane method and apparatus for separating isotopic water constituents from light water. The method involves providing a supported membrane of an aromatic polyphosphazene and pressurizing the water on one side of the membrane thereby forcing the light water through the supported membrane while isotopic water constituents are retained or vice versa. The apparatus of the present invention includes an aromatic polyphosphazene placed on a porous support and means for pressurizing water through the membrane while certain isotopic water constituents are retained.

  13. A rapid and versatile method to label receptor ligands using "click" chemistry: Validation with the muscarinic M1 antagonist pirenzepine.

    PubMed

    Bonnet, Dominique; Ilien, Brigitte; Galzi, Jean-Luc; Riché, Stéphanie; Antheaune, Cyril; Hibert, Marcel

    2006-01-01

    Tagged biologically active molecules represent powerful pharmacological tools to study and characterize ligand-receptor interactions. However, the labeling of such molecules is not trivial, especially when poorly soluble tags have to be incorporated. The classical method of coupling usually necessitates a tedious final purification step to remove the excess of reagents and to isolate tagged molecules. To overcome this limitation, Cu(I)-catalyzed 1,3-dipolar cycloaddition, referred to as "click" chemistry, was evaluated as a tool to facilitate the access to labeled molecules. In order to validate the approach, we focused our attention on the incorporation of a fluorophore (Lissamine Rhodamine B), a nonfluorescent dye (Patent Blue VF), or biotin into a muscarinic antagonist scaffold derived from pirenzepine. The reaction performed in acetonitrile/water, in the presence of CuSO4 and Cu wire, allowed us to obtain three novel pirenzepine derivatives with high purity and in good yield. No coupling reagents were needed, and the quasi-stoichiometric conditions of the reaction enabled the straightforward isolation of the final product by simple precipitation and its use in bioassays. The affinity of the compounds for the human M1 muscarinic receptor fused to EGFP was checked under classical radioligand and FRET binding conditions. The three pirenzepine constructs display a nanomolar affinity for the M1 receptor. In addition, both dye-labeled derivatives behave as potent acceptors of energy from excited EGFP with a very high quenching efficiency.

  14. Adaboost-LLP: A Boosting Method for Learning With Label Proportions.

    PubMed

    Qi, Zhiquan; Meng, Fan; Tian, Yingjie; Niu, Lingfeng; Shi, Yong; Zhang, Peng

    2017-08-15

    How to solve the classification problem with only label proportions has recently drawn increasing attention in the machine learning field. In this paper, we propose an ensemble learning strategy to deal with the learning problem with label proportions (LLP). In detail, we first give a loss function based on different weights for LLP, and then construct the corresponding weak classifier, at the same time, estimate its conditional probabilities by a standard logistic function. At last, by introducing the maximum likelihood estimation, we propose a new anyboost learning system for LLP (called Adaboost-LLP). Unlike traditional methods, our method does not make any restrictive assumptions on training set; at the same time, compared with alter-∝SVM, Adaboost-LLP exploits more extra weight information and uses multiple weak classifiers that can be solved efficiently to combine a strong classifier. All experiments show that our method outperforms the existing methods in both accuracy and training time.

  15. Gluten and gluten-free: issues and considerations of labeling regulations, detection methods, and assay validation.

    PubMed

    Diaz-Amigo, Carmen; Popping, Bert

    2012-01-01

    Gluten is a commonly used cereal derivative found in bakery products, among other items. In some susceptible individuals, however, it triggers immune responses of different kinds; there is, to a lesser extent, the wheat allergy that is immunoglobulin E (IgE)-mediated and leads to histamine release and typical allergic symptoms. In this case, other water-soluble proteins, like albumins, are also involved. On the other hand, there is, more frequently, celiac disease (CD), where the gluten causes immune reactions in the intestines of certain individuals, leading to degeneration of villi, which typically leads to malabsorption of nutrients and, consequently, malnutrition. The only currently effective health strategy for affected consumers is avoidance of gluten-containing products, based on clear labeling rules. However, despite unanimously accepted Codex definitions by all member jurisdictions, the national implementation of equivalent laws shows significant differences. In the context of CD and in support of the gluten-free statement, regulatory enforcement, as well as manufacturers' quality controls are mostly based on analytical results. However, numerous methods are available, some of which have been validated better than others, and many provide different results on identical samples. Reasons include detection of different gluten components and variability in extraction efficiency due to different buffer compositions, especially from processed foods. Last but not least, the lack of reference materials is hindering the process of generating comparable data across different ELISA kits, as well as other methods. How can such data still be used to support a gluten-free claim? New methodologies, in particular mass spectrometric analysis of gluten derived peptides, are being introduced in numerous laboratories. This methodology is not only capable of detecting gluten derived peptides but can also differentiate between and quantitate wheat, barley, rye, and oat. This

  16. A method to trace root-respired CO2 using a 13C label

    NASA Astrophysics Data System (ADS)

    Cooperdock, S.; Breecker, D.; Litvak, M. E.

    2014-12-01

    In order to partition total soil respiration into root respiration and decomposition under ambient conditions in desert soils, the following method was developed using 13C-labeled CO2 in a modern juniper savannah in central New Mexico. The labeled CO2 was mixed with ambient air and pumped into a small (2.5 m diameter and 1.4 m tall) juniper tree canopy . 10 L of the 13CO2 was sufficient to generate a stream of air at 20 L/min for 1 hour with a CO2 concentration of 540 ppm and a δ13C value of approximately 35,000‰. Plastic tarpaulins were used as a wind block. The 13CO2 -labeled air was applied to the canopy during peak photosynthesis between 10 and 11 am on June 30 2014 during which canopy air CO2 was elevated by approximately 10 ppm over ambient and had δ13C values ranging from 50 to 1000 ‰. Over the next three days, gas and tissue samples were collected in order to trace the 13C label through the juniper tree. Leaf and root samples collected from the labeled tree and from several control trees were loaded into exetainer vials, flushed with CO2-free air and incubated in the dark for 5 hours in order to measure the carbon isotope composition of respired CO2. Samples of soil pore space gas were collected from wells under the labeled tree and a control tree and were transported to the laboratory in He-flushed exetainer vials. The δ13C values of CO2 in the soil gas samples and in the headspace of incubation vials were measured using an isotope ratio mass spectrometer. The δ13C values of foliar respiration were significantly higher than those of the control (by 3.6‰, p < 0.01) one and two days after labeling and δ13C values of root-respired CO2 were significantly higher (by 0.7‰, p = 0.01) than those of the control three days after labeling. In addition, δ13C values of soil respired CO2, determined from measurements of soil pore space CO2 at 50 cm three days after labeling, were significantly higher (by 0.7‰, p < 0.03)) for the labeled tree than control

  17. Method of treating waste water

    DOEpatents

    Deininger, James P.; Chatfield, Linda K.

    1995-01-01

    A process of treating water to remove metal ion contaminants contained therein, said metal ion contaminants selected from the group consisting of metals in Groups 8, 1b, 2b, 4a, 5a, or 6a of the periodic table, lanthanide metals, and actinide metals including transuranic element metals, by adjusting the pH of a metal ion contaminant-containing water source to within the range of about 6.5 to about 14.0, admixing the water source with a mixture of an alkali or alkaline earth ferrate and a water soluble salt, e.g., a zirconium salt, in an amount sufficient to form a precipitate within the water source, the amount the mixture of ferrate and water soluble salt effective to reduce the metal ion contaminant concentration in the water source, permitting the precipitate in the admixture to separate and thereby yield a supernatant liquid having a reduced metal ion contaminant concentration, and separating the supernatant liquid having the reduced metal ion contaminant concentration from the admixture is provided. A composition of matter including an alkali or alkaline earth ferrate and a water soluble salt, e.g., a zirconium salt, is also provided.

  18. A novel decision-tree method for structured continuous-label classification.

    PubMed

    Hu, Hsiao-Wei; Chen, Yen-Liang; Tang, Kwei

    2013-12-01

    Structured continuous-label classification is a variety of classification in which the label is continuous in the data, but the goal is to classify data into classes that are a set of predefined ranges and can be organized in a hierarchy. In the hierarchy, the ranges at the lower levels are more specific and inherently more difficult to predict, whereas the ranges at the upper levels are less specific and inherently easier to predict. Therefore, both prediction specificity and prediction accuracy must be considered when building a decision tree (DT) from this kind of data. This paper proposes a novel classification algorithm for learning DT classifiers from data with structured continuous labels. This approach considers the distribution of labels throughout the hierarchical structure during the construction of trees without requiring discretization in the preprocessing stage. We compared the results of the proposed method with those of the C4.5 algorithm using eight real data sets. The empirical results indicate that the proposed method outperforms the C4.5 algorithm with regard to prediction accuracy, prediction specificity, and computational complexity.

  19. A method for following human lymphocyte traffic using indium-111 oxine labelling.

    PubMed Central

    Wagstaff, J; Gibson, C; Thatcher, N; Ford, W L; Sharma, H; Benson, W; Crowther, D

    1981-01-01

    A method is described whereby large numbers of human lymphocytes are separated from peripheral blood and labelled in vitro with indium-111 oxine. Following autologous reinjection, the distribution within the body is followed by means of serial blood samples, surface-probe counting and gamma camera imaging. The distribution of radioactivity following reinjection of heat-damaged labelled lymphocytes and free indium-111 oxine is different from that of 'normal' lymphocytes. The results suggest that the separation and labelling procedure does not cause significant physical damage to the lymphocytes The importance of restricting the specific lymphocyte activity to 20-40 microCi per 10(8) cells in order to minimize radiation damage to the lymphocytes is emphasized. Good resolution of lymphoid structures is obtained using gamma camera imaging and the changes recorded in organ distribution correlate well with data from animal models of lymphocyte migration. Thus, indium-111 oxine labelling of human lymphocytes provides a non-invasive method whereby the migratory properties of human lymphocytes can be followed. Images Fig. 2 Fig. 3 Fig. 4 PMID:7285387

  20. Synthesis of water-dispersible zinc oxide quantum dots with antibacterial activity and low cytotoxicity for cell labeling

    NASA Astrophysics Data System (ADS)

    Hsu, Shan-hui; Lin, Ying Yi; Huang, Sherry; Lem, Kwok Wai; Huong Nguyen, Dinh; Lee, Dai Soo

    2013-11-01

    Typical photoluminescent semiconductor nanoparticles, called quantum dots (QDs), have potential applications in biological labeling. When used to label stem cells, QDs may impair the differentiation capacity of the stem cells. In this study, we synthesized zinc oxide (ZnO) QDs in methanol with an average size of ∼2 nm. We then employed two different types of polyethylene glycol (PEG) molecules (SH-PEG-NH2 and NH2-PEG-NH2) to conjugate ZnO QDs and made them water-dispersible. Fourier transform infrared spectroscopy spectra indicated the attachment of PEG molecules on ZnO QDs. No obvious size alteration was observed for ZnO QDs after PEG conjugation. The water-dispersible ZnO QDs still retained the antibacterial activity and fluorescence intensity. The cytotoxicity evaluation revealed that ZnO QDs at higher concentrations decreased cell viability but were generally safe at 30 ppm or below. Cell lines of hepatocytes (HepG2), osteoblasts (MC3T3-E1) and mesenchymal stem cells (MSCs) were successfully labeled by the water-dispersible ZnO QDs at 30 ppm. The ZnO QD-labeled MSCs maintained their stemness and differentiation capacity. Therefore, we conclude that the water-dispersible ZnO QDs developed in this study have antibacterial activity, low cytotoxicity, and proper labeling efficiency, and can be used to label a variety of cells including stem cells.

  1. A method to determine plant water source using transpired water

    NASA Astrophysics Data System (ADS)

    Menchaca, L. B.; Smith, B. M.; Connolly, J.; Conrad, M.; Emmett, B.

    2007-04-01

    A method to determine the stable isotope ratio of a plant's water source using the plant's transpired water is proposed as an alternative to standard xylem extraction methods. The method consists of periodically sampling transpired waters from shoots or leaves enclosed in sealed, transparent bags which create a saturated environment, preclude further evaporation and allow the progressive mixing of evaporated transpired water and un-evaporated xylem water. The method was applied on trees and shrubs coexisting in a non-irrigated area where stable isotope ratios of local environmental waters are well characterized. The results show Eucalyptus globulus (tree) and Genista monspessulana (shrub) using water sources of different isotopic ratios congruent with groundwater and soil water respectively. In addition, tritium concentrations indicate that pine trees (Pinus sylvestris) switch water source from soil water in the winter to groundwater in the summer. The method proposed is particularly useful in remote or protected areas and in large scale studies related to water management, environmental compliance and surveillance, because it eliminates the need for destructive sampling and greatly reduces costs associated with laboratory extraction of xylem waters from plant tissues for isotopic analyses.

  2. Method of treating waste water

    DOEpatents

    Deininger, J. Paul; Chatfield, Linda K.

    1991-01-01

    A process of treating water to remove transuranic elements contained therein by adjusting the pH of a transuranic element-containing water source to within the range of about 6.5 to about 14.0, admixing the water source with an alkali or alkaline earth ferrate in an amount sufficient to form a precipitate within the water source, the amount of ferrate effective to reduce the transuranic element concentration in the water source, permitting the precipitate in the admixture to separate and thereby yield a supernatant liquid having a reduced transuranic element concentration, and separating the supernatant liquid having the reduced transuranic element concentration from the admixture is provided. Additionally, a water soluble salt, e.g., a zirconium salt, can be added with the alkali or alkaline earth ferrate in the process to provide greater removal efficiencies. A composition of matter including an alkali or alkaline earth ferrate and a water soluble salt, e.g., a zirconium salt, is also provided.

  3. Liquid state DNP for water accessibility measurements on spin-labeled membrane proteins at physiological temperatures

    NASA Astrophysics Data System (ADS)

    Doll, Andrin; Bordignon, Enrica; Joseph, Benesh; Tschaggelar, René; Jeschke, Gunnar

    2012-09-01

    We demonstrate the application of continuous wave dynamic nuclear polarization (DNP) at 0.35 T for site-specific water accessibility studies on spin-labeled membrane proteins at concentrations in the 10-100 μM range. The DNP effects at such low concentrations are weak and the experimentally achievable dynamic nuclear polarizations can be below the equilibrium polarization. This sensitivity problem is solved with an optimized home-built DNP probe head consisting of a dielectric microwave resonator and a saddle coil as close as possible to the sample. The performance of the probe head is demonstrated with both a modified pulsed EPR spectrometer and a dedicated CW EPR spectrometer equipped with a commercial NMR console. In comparison to a commercial pulsed ENDOR resonator, the home-built resonator has an FID detection sensitivity improvement of 2.15 and an electron spin excitation field improvement of 1.2. The reproducibility of the DNP results is tested on the water soluble maltose binding protein MalE of the ABC maltose importer, where we determine a net standard deviation of 9% in the primary DNP data in the concentration range between 10 and 100 μM. DNP parameters are measured in a spin-labeled membrane protein, namely the vitamin B12 importer BtuCD in both detergent-solubilized and reconstituted states. The data obtained in different nucleotide states in the presence and absence of binding protein BtuF reveal the applicability of this technique to qualitatively extract water accessibility changes between different conformations by the ratio of primary DNP parameters ɛ. The ɛ-ratio unveils the physiologically relevant transmembrane communication in the transporter in terms of changes in water accessibility at the cytoplasmic gate of the protein induced by both BtuF binding at the periplasmic region of the transporter and ATP binding at the cytoplasmic nucleotide binding domains.

  4. Analysis of Hydrogen Isotopic Exchange: Lava Creek Tuff Ash and Isotopically Labeled Water

    NASA Astrophysics Data System (ADS)

    Ross, A. M.; Seligman, A. N.; Bindeman, I. N.; Nolan, G. S.

    2015-12-01

    Nolan and Bindeman (2013) placed secondarily hydrated ash from the 7.7 ka eruption of Mt. Mazama (δD=-149‰, 2.3wt% H2Ot) in isotopically labeled water (+650 ‰ δD, +56 ‰ δ18O) and observed that the H2Ot and δ18O values remained constant, but the δD values of ash increased with the surrounding water at 20, 40 and 70 °C. We expand on this work by conducting a similar experiment with ash from the 640 ka Lava Creek Tuff (LCT, δD of -128 ‰; 2.1 wt.% H2Ot) eruption of Yellowstone to see if significantly older glass (with a hypothesized gel layer on the surface shielding the interior from alteration) produces the same results. We have experiments running at 70, 24, and 5 °C, and periodically remove ~1.5 mg of glass to measure the δD (‰) and H2Ot (wt.%) of water extracted from the glass on a TC/EA MAT 253 continuous flow system. After 600 hours, the δD of the samples left at 5 and 24 °C remains at -128 ‰, but increased 8‰ for the 70 °C run series. However, there is no measurable change in wt.% of H2Ot, indicating that hydrogen exchange is not dictated by the addition of water. We are measuring and will report further progress of isotope exchange. We also plan to analyze the water in the LCT glass for δ18O (‰) to see if, as is the case for the Mt. Mazama glass, the δ18O (‰) remains constant. We also analyzed Mt. Mazama glass from the Nolan and Bindeman (2013) experiments that have now been sitting in isotopically labeled water at room temperature for ~5 years. The water concentration is still unchanged (2.3 wt.% H2Ot), and the δD of the water in the glass is now -111 ‰, causing an increase of 38 ‰. Our preliminary results show that exchange of hydrogen isotopes of hydrated glass is not limited by the age of the glass, and that the testing of hydrogen isotopes of secondarily hydrated glass, regardless of age, may not be a reliable paleoclimate indicator.

  5. Neuronal Tracing with Magnetic Labels: NMR Imaging Methods, Preliminary Results, and New Optimized Coils.

    NASA Astrophysics Data System (ADS)

    Ghosh, Pratik

    1992-01-01

    The investigations focussed on in vivo NMR imaging studies of magnetic particles with and within neural cells. NMR imaging methods, both Fourier transform and projection reconstruction, were implemented and new protocols were developed to perform "Neuronal Tracing with Magnetic Labels" on small animal brains. Having performed the preliminary experiments with neuronal tracing, new optimized coils and experimental set-up were devised. A novel gradient coil technology along with new rf-coils were implemented, and optimized for future use with small animals in them. A new magnetic labelling procedure was developed that allowed labelling of billions of cells with ultra -small magnetite particles in a short time. The relationships among the viability of such cells, the amount of label and the contrast in the images were studied as quantitatively as possible. Intracerebral grafting of magnetite labelled fetal rat brain cells made it possible for the first time to attempt monitoring in vivo the survival, differentiation, and possible migration of both host and grafted cells in the host rat brain. This constituted the early steps toward future experiments that may lead to the monitoring of human brain grafts of fetal brain cells. Preliminary experiments with direct injection of horse radish peroxidase-conjugated magnetite particles into neurons, followed by NMR imaging, revealed a possible non-invasive alternative, allowing serial study of the dynamic transport pattern of tracers in single living animals. New gradient coils were built by using parallel solid-conductor ribbon cables that could be wrapped easily and quickly. Rapid rise times provided by these coils allowed implementation of fast imaging methods. Optimized rf-coil circuit development made it possible to understand better the sample-coil properties and the associated trade -offs in cases of small but conducting samples.

  6. Preparation of lucifer yellow fluorescent liposomes: A method for cells' membrane labeling.

    PubMed

    Menna, C; Calonghi, N; Masotti, L; Neyroz, P

    1993-03-01

    This report describes a method to conjugate lucifer yellow to the external surface of liposomes. The heterobifunctional cross-linking reagentN-succinimidyl 3-(2-pyridyldithio)propionate has been used to activate DMPE molecules. The DMPE-dithiopyridine product has been mixed with DMPC to prepare liposome vesicles. These have been reduced by DTT and finally reacted with lucifer yellow-iodoacetamide to produce the fluorescence-labeled vesicles. The quenching of their fluorescence intensity by Kl is consistent with fully exposed fluorophores. The decay of the fluorescence intensity of the lipid-bound lucifer yellow is biexponential (τ1=7.9 ns; τ2=1.1 ns), with a relative yield of 0.16. When the fluorescent liposomes are mixed with cells, the lucifer yellow-DMPE derivative is transferred. Boar spermatozoa and peripheral human blood lymphocytes have been used as cellular models. The extent of incorporation is dependent on the incubation time and temperature. At 36°C, lucifer yellow fluorescence appears in the spermatozoa cells after 10 min of incubation and reaches its maximum at about 60 min. The fluorescent phospholipid derivative seems to incorporate specifically into membrane structures. The highest labeling ratio is observed with integer, scarcely motile, spermatozoa. A poorer labeling yield (≈15%) is found with lymphocytes. Interestingly, photobleaching due to epiillumination of the labeled cells is apparently negligible and cells are clearly visible after irradiation times ranging from several minutes to few hours.

  7. An optimized method for measuring fatty acids and cholesterol in stable isotope-labeled cells.

    PubMed

    Argus, Joseph P; Yu, Amy K; Wang, Eric S; Williams, Kevin J; Bensinger, Steven J

    2017-02-01

    Stable isotope labeling has become an important methodology for determining lipid metabolic parameters of normal and neoplastic cells. Conventional methods for fatty acid and cholesterol analysis have one or more issues that limit their utility for in vitro stable isotope-labeling studies. To address this, we developed a method optimized for measuring both fatty acids and cholesterol from small numbers of stable isotope-labeled cultured cells. We demonstrate quantitative derivatization and extraction of fatty acids from a wide range of lipid classes using this approach. Importantly, cholesterol is also recovered, albeit at a modestly lower yield, affording the opportunity to quantitate both cholesterol and fatty acids from the same sample. Although we find that background contamination can interfere with quantitation of certain fatty acids in low amounts of starting material, our data indicate that this optimized method can be used to accurately measure mass isotopomer distributions for cholesterol and many fatty acids isolated from small numbers of cultured cells. Application of this method will facilitate acquisition of lipid parameters required for quantifying flux and provide a better understanding of how lipid metabolism influences cellular function. Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.

  8. A Low-Storage-Consumption XML Labeling Method for Efficient Structural Information Extraction

    NASA Astrophysics Data System (ADS)

    Liang, Wenxin; Takahashi, Akihiro; Yokota, Haruo

    Recently, labeling methods to extract and reconstruct the structural information of XML data, which are important for many applications such as XPath query and keyword search, are becoming more attractive. To achieve efficient structural information extraction, in this paper we propose C-DO-VLEI code, a novel update-friendly bit-vector encoding scheme, based on register-length bit operations combining with the properties of Dewey Order numbers, which cannot be implemented in other relevant existing schemes such as ORDPATH. Meanwhile, the proposed method also achieves lower storage consumption because it does not require either prefix schema or any reserved codes for node insertion. We performed experiments to evaluate and compare the performance and storage consumption of the proposed method with those of the ORDPATH method. Experimental results show that the execution times for extracting depth information and parent node labels using the C-DO-VLEI code are about 25% and 15% less, respectively, and the average label size using the C-DO-VLEI code is about 24% smaller, comparing with ORDPATH.

  9. Development of methods to measure virus inactivation in fresh waters.

    PubMed Central

    Ward, R L; Winston, P E

    1985-01-01

    This study concerns the identification and correction of deficiencies in methods used to measure inactivation rates of enteric viruses seeded into environmental waters. It was found that viable microorganisms in an environmental water sample increased greatly after addition of small amounts of nutrients normally present in the unpurified seed virus preparation. This burst of microbial growth was not observed after seeding the water with purified virus. The use of radioactively labeled poliovirus revealed that high percentages of virus particles, sometimes greater than 99%, were lost through adherence to containers, especially in less turbid waters. This effect was partially overcome by the use of polypropylene containers and by the absence of movement during incubation. Adherence to containers clearly demonstrated the need for labeled viruses to monitor losses in this type of study. Loss of viral infectivity in samples found to occur during freezing was avoided by addition of broth. Finally, microbial contamination of the cell cultures during infectivity assays was overcome by the use of gentamicin and increased concentrations of penicillin, streptomycin, and amphotericin B. PMID:3004328

  10. Ionomer-Membrane Water Processing Methods

    NASA Technical Reports Server (NTRS)

    MacCallum, Taber K. (Inventor); Kelsey, Laura (Inventor)

    2016-01-01

    This disclosure provides water processing apparatuses, systems, and methods for recovering water from wastewater such as urine. The water processing apparatuses, systems, and methods can utilize membrane technology for extracting purified water in a single step. A containment unit can include an ionomer membrane, such as Nafion(TradeMark) over a hydrophobic microporous membrane, such as polytetrafluoroethylene (PTFE). The containment unit can be filled with wastewater, and the hydrophobic microporous membrane can be impermeable to liquids and solids of the wastewater but permeable to gases and vapors of the wastewater, and the ionomer membrane can be permeable to water vapor but impermeable to one or more contaminants of the gases and vapors. The containment unit can be exposed to a dry purge gas to maintain a water vapor partial pressure differential to drive permeation of the water vapor, and the water vapor can be collected and processed into potable water.

  11. A new powerful association for immunoassay: organometallic label conjugated to solvent separation method.

    PubMed

    Mariet, F; Brossier, P

    1990-05-01

    The convenient use of solvents as a method to separate free (F) and bound (B) fractions in drug immunoassays which utilize organometallic moieties as labels, has been developed. Two types of drugs labelled with an organometallic complex and denominated metallohaptens (or metallotracers) are studied as model assays: desipramine (tricyclic antidepressant) and phenobarbital (antiepileptic agent). Different organic solvents have potentialities to extract these metallohaptens. To illustrate this solvent procedure we describe antiserum dilution curves and we compare the results with those obtained for the same samples using radiotracers in conjunction with dextran coated charcoal as separation agent. This immunoassay method which associates a metallotracer and B/F solvent separation appears to be fast, cheap, without the step of centrifugation.

  12. Enantiomer labelling, a method for the quantitative analysis of amino acids.

    PubMed

    Frank, H; Nicholson, G J; Bayer, E

    1978-12-21

    Enantiomer labelling a method for the quntitative analysis of optically active natural compounds by gas chromatography, involves the use of the unnatural enantiomer as an internal standard. With Chirasil-Val, a chiral stationary phase that is thermally stable up to up to 240 degrees, the enantiomers of amino acids and a variety of other compounds can be separated and quantitated. Incomplete recovery from the sample, incomplete derivatization, hydrolysis and thermal decomposition of the derivative and shifting response factors can be compensated for by adding the unnatural enantiomer. The accuracy of amino acid analysis by enantiomer labelling is equal or superior to that of hitherto known methods. The procedure affords a complete analysis of peptides with respect to both amino acid composition and the optical purity of each amino acid.

  13. Radiation-Inactivation of Meat Proteases as Determined by a C-14-Labeled Hemoglobin Method

    DTIC Science & Technology

    1975-11-01

    AD i TECHNICAL REPORT I TR-76-33FEL RADIATION-INACTIVATION OF MEAT PROTEASES AS DETERMINED BY A 14C- LABELED HEMOGLOBIN METHOD Irradiated Food ...Products Group ■ Radiation Preservation of Food Division Approved for public release; distribution unlimited. November 1975 UNITED STATES ARMY...NATICK RESEARCH and DEVELOPMENT COMMAND NATICK, MASSACHUSETTS 01760 Food Engineering Laboratory FEL-48 Approved for public release; distribution

  14. Transport of North Pacific 137Cs labeled waters to the south-eastern Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Sanchez-Cabeza, J. A.; Levy, I.; Gastaud, J.; Eriksson, M.; Osvath, I.; Aoyama, M.; Povinec, P. P.; Komura, K.

    2011-04-01

    During the reoccupation of the WOCE transect A10 at 30°S by the BEAGLE2003 cruise, the SHOTS project partners collected a large number of samples for the analysis of isotopic tracers. 137Cs was mostly deposited on the oceans surface during the late 1950s and early 1960s, after the atmospheric detonation of large nuclear devices, which mostly occurred in the Northern Hemisphere. The development of advanced radioanalytical and counting techniques allowed to obtain, for the first time in this region, a zonal section of 137Cs water concentrations, where little information existed before, thus constituting an important benchmark for further studies. 137Cs concentrations in the upper waters (0-1000 m) of the south-eastern Atlantic Ocean are similar to those observed in the south-western Indian Ocean, suggesting transport of 137Cs labeled waters by the Agulhas current to the Benguela Current region. In contrast, bomb radiocarbon data do not show this feature, indicating the usefulness of 137Cs as a radiotracer of water mass transport from the Indian to the South Atlantic Ocean.

  15. Egg labeling methods for gastric emptying scintigraphy are not equivalent in producing a stable solid meal.

    PubMed

    Knight, Linda C; Kantor, Steven; Doma, Siva; Parkman, Henry P; Maurer, Alan H

    2007-11-01

    A wide range of radiolabeled test meals have been used for gastric emptying scintigraphy. The purpose of this study was to test whether (99m)Tc-sulfur colloid-labeled liquid egg white is as stable as 2 fresh whole eggs labeled with (99m)Tc-sulfur colloid and whether the cooking method is important. Whole eggs and liquid egg white were mixed with (99m)Tc-sulfur colloid and cooked by either microwaving or frying on a griddle. The cooked eggs were tested for breakdown after 2 and 4 h of incubation in gastric fluid or HCl. Labeled liquid egg white, prepared by either method of cooking, exhibited less breakdown in gastric fluid than whole eggs. Whole eggs cooked in the microwave exhibited significantly more breakdown than liquid egg white. (99m)Tc-Sulfur colloid binds better to egg whites compared with whole eggs. These results emphasize the need to evaluate the stability of new radiolabeled test meal preparations, including the method of cooking.

  16. SVM model for quality assessment of medium resolution mass spectra from 18O-water labeling experiments.

    PubMed

    Nefedov, Alexey V; Gilski, Miroslaw J; Sadygov, Rovshan G

    2011-04-01

    We describe a method for assessing the quality of mass spectra and improving reliability of relative ratio estimations from (18)O-water labeling experiments acquired from low resolution mass spectrometers. The mass profiles of heavy and light peptide pairs are often affected by artifacts, including coeluting contaminant species, noise signal, instrumental fluctuations in measuring ion position and abundance levels. Such artifacts distort the profiles, leading to erroneous ratio estimations, thus reducing the reliability of ratio estimations in high throughput quantification experiments. We used support vector machines (SVMs) to filter out mass spectra that deviated significantly from expected theoretical isotope distributions. We built an SVM classifier with a decision function that assigns a score to every mass profile based on such spectral features as mass accuracy, signal-to-noise ratio, and differences between experimental and theoretical isotopic distributions. The classifier was trained using a data set obtained from samples of mouse renal cortex. We then tested it on protein samples (bovine serum albumin) mixed in five different ratios of labeled and unlabeled species. We demonstrated that filtering the data using our SVM classifier results in as much as a 9-fold reduction in the coefficient of variance of peptide ratios, thus significantly improving the reliability of ratio estimations.

  17. A calorimetric method to determine water activity

    NASA Astrophysics Data System (ADS)

    Björklund, Sebastian; Wadsö, Lars

    2011-11-01

    A calorimetric method to determine water activity covering the full range of the water activity scale is presented. A dry stream of nitrogen gas is passed either over the solution whose activity should be determined or left dry before it is saturated by bubbling through water in an isothermal calorimeter. The unknown activity is in principle determined by comparing the thermal power of vaporization related to the gas stream with unknown activity to that with zero activity. Except for three minor corrections (for pressure drop, non-perfect humidification, and evaporative cooling) the unknown water activity is calculated solely based on the water activity end-points zero and unity. Thus, there is no need for calibration with references with known water activities. The method has been evaluated at 30 °C by measuring the water activity of seven aqueous sodium chloride solutions ranging from 0.1 mol kg-1 to 3 mol kg-1 and seven saturated aqueous salt solutions (LiCl, MgCl2, NaBr, NaCl, KCl, KNO3, and K2SO4) with known water activities. The performance of the method was adequate over the complete water activity scale. At high water activities the performance was excellent, which is encouraging as many other methods used for water activity determination have limited performance at high water activities.

  18. Label-Free Electrical Immunosensor for Highly Sensitive and Specific Detection of Microcystin-LR in Water Samples.

    PubMed

    Tan, Feng; Saucedo, Nuvia Maria; Ramnani, Pankaj; Mulchandani, Ashok

    2015-08-04

    Microcystin-LR (MCLR) is one of the most commonly detected and toxic cyclic heptapeptide cyanotoxins released by cyanobacterial blooms in surface waters, for which sensitive and specific detection methods are necessary to carry out its recognition and quantification. Here, we present a single-walled carbon nanotube (SWCNTs)-based label-free chemiresistive immunosensor for highly sensitive and specific detection of MCLR in different source waters. MCLR was initially immobilized on SWCNTs modified interdigitated electrode, followed by incubation with monoclonal anti-MCLR antibody. The competitive binding of MCLR in sample solutions induced departure of the antibody from the antibody-antigen complexes formed on SWCNTs, resulting in change in the conductivity between source and drain of the sensor. The displacement assay greatly improved the sensitivity of the sensor compared with direct immunoassay on the same device. The immunosensor exhibited a wide linear response to log value of MCLR concentration ranging from 1 to 1000 ng/L, with a detection limit of 0.6 ng/L. This method showed good reproducibility, stability and recovery. The proposed method provides a powerful tool for rapid and sensitive monitoring of MCLR in environmental samples.

  19. Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids

    NASA Technical Reports Server (NTRS)

    Ni, J.; Lipert, R. J.; Dawson, G. B.; Porter, M. D.

    1999-01-01

    An immunoassay readout method based on surface-enhanced Raman scattering (SERS) is described. The method exploits the SERS-derived signal from reporter molecules that are coimmobilized with biospecific species on gold colloids. This concept is demonstrated in a dualanalyte sandwich assay, in which two different antibodies covalently bound to a solid substrate specifically capture two different antigens from an aqueous sample. The captured antigens in turn bind selectively to their corresponding detection antibodies. The detection antibodies are conjugated with gold colloids that are labeled with different Raman reporter molecules, which serve as extrinsic labels for each type of antibody. The presence of a specific antigen is established by the characteristic SERS spectrum of the reporter molecule. A near-infrared diode laser was used to excite efficiently the SERS signal while minimizing fluorescence interference. We show that, by using different labels with little spectral overlap, two different antigenic species can be detected simultaneously. The potential of this concept to function as a readout strategy for multiple analytes is briefly discussed.

  20. Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids

    NASA Technical Reports Server (NTRS)

    Ni, J.; Lipert, R. J.; Dawson, G. B.; Porter, M. D.

    1999-01-01

    An immunoassay readout method based on surface-enhanced Raman scattering (SERS) is described. The method exploits the SERS-derived signal from reporter molecules that are coimmobilized with biospecific species on gold colloids. This concept is demonstrated in a dualanalyte sandwich assay, in which two different antibodies covalently bound to a solid substrate specifically capture two different antigens from an aqueous sample. The captured antigens in turn bind selectively to their corresponding detection antibodies. The detection antibodies are conjugated with gold colloids that are labeled with different Raman reporter molecules, which serve as extrinsic labels for each type of antibody. The presence of a specific antigen is established by the characteristic SERS spectrum of the reporter molecule. A near-infrared diode laser was used to excite efficiently the SERS signal while minimizing fluorescence interference. We show that, by using different labels with little spectral overlap, two different antigenic species can be detected simultaneously. The potential of this concept to function as a readout strategy for multiple analytes is briefly discussed.

  1. Apparatus and method for reading two-dimensional electrophoretograms containing .beta.-ray-emitting labeled compounds

    DOEpatents

    Anderson, Herbert L.; Kinnison, W. Wayne; Lillberg, John W.

    1987-01-01

    Apparatus and method for electronically reading planar two dimensional .beta.-ray emitter-labeled gel electrophoretograms. A single, flat rectangular multiwire proportional chamber is placed in close proximity to the gel and the assembly placed in an intense uniform magnetic field disposed in a perpendicular manner to the rectangular face of the proportional chamber. Beta rays emitted in the direction of the proportional chamber are caused to execute helical motions which substantially preserve knowledge of the coordinates of their origin in the gel. Perpendicularly oriented, parallel wire, parallel plane cathodes electronically sense the location of the .beta.-rays from ionization generated thereby in a detection gas coupled with an electron avalanche effect resulting from the action of a parallel wire anode located therebetween. A scintillator permits the present apparatus to be rendered insensitive when signals are generated from cosmic rays incident on the proportional chamber. Resolution for concentrations of radioactive compounds in the gel exceeds 700 .mu.m. The apparatus and method of the present invention represent a significant improvement over conventional autoradiographic techniques in dynamic range, linearity and sensitivity of data collection. A concentration and position map for gel electrophoretograms having significant concentrations of labeled compounds and/or highly radioactive labeling nuclides can generally be obtained in less than one hour.

  2. Quantitative study on the fate of residual soil nitrate in winter wheat based on a 15N-labeling method

    PubMed Central

    Zhang, Jing-Ting; Wang, Zhi-Min; Liang, Shuang-Bo; Zhang, Ying-Hua; Lu, Lai-Qing; Wang, Run-Zheng

    2017-01-01

    A considerable amount of surplus nitrogen (N), which primarily takes the form of nitrate, accumulates in the soil profile after harvesting crops from an intensive production system in the North China Plain. The residual soil nitrate (RSN) is a key factor that is included in the N recommendation algorithm. Quantifying the utilization and losses of RSN is a fundamental necessity for optimizing crop N management, improving N use efficiency, and reducing the impact derived from farmland N losses on the environment. In this study, a 15N-labeling method was introduced to study the fate of the RSN quantitatively during the winter wheat growing season by 15N tracer technique combined with a soil column study. A soil column with a 2 m height was vertically divided into 10 20-cm layers, and the RSN in each layer was individually labeled with a 15N tracer before the wheat was sown. The results indicated that approximately 17.68% of the crop N derived from RSN was located in the 0–2 m soil profile prior to wheat sowing. The wheat recovery proportions of RSN at various layers ranged from 0.21% to 33.46%. The percentages that still remained in the soil profile after the wheat harvest ranged from 47.08% to 75.44%, and 19.46–32.64% of the RSN was unaccounted for. Upward and downward movements in the RSN were observed, and the maximum upward and downward distances were 40 cm and 100 cm, respectively. In general, the 15N-labeling method contributes to a deeper understanding of the fates of the RSN. Considering the low crop recovery of the RSN from deep soil layers, water and N saving practices should be adopted during crop production. PMID:28170440

  3. Quantitative study on the fate of residual soil nitrate in winter wheat based on a 15N-labeling method.

    PubMed

    Zhang, Jing-Ting; Wang, Zhi-Min; Liang, Shuang-Bo; Zhang, Ying-Hua; Zhou, Shun-Li; Lu, Lai-Qing; Wang, Run-Zheng

    2017-01-01

    A considerable amount of surplus nitrogen (N), which primarily takes the form of nitrate, accumulates in the soil profile after harvesting crops from an intensive production system in the North China Plain. The residual soil nitrate (RSN) is a key factor that is included in the N recommendation algorithm. Quantifying the utilization and losses of RSN is a fundamental necessity for optimizing crop N management, improving N use efficiency, and reducing the impact derived from farmland N losses on the environment. In this study, a 15N-labeling method was introduced to study the fate of the RSN quantitatively during the winter wheat growing season by 15N tracer technique combined with a soil column study. A soil column with a 2 m height was vertically divided into 10 20-cm layers, and the RSN in each layer was individually labeled with a 15N tracer before the wheat was sown. The results indicated that approximately 17.68% of the crop N derived from RSN was located in the 0-2 m soil profile prior to wheat sowing. The wheat recovery proportions of RSN at various layers ranged from 0.21% to 33.46%. The percentages that still remained in the soil profile after the wheat harvest ranged from 47.08% to 75.44%, and 19.46-32.64% of the RSN was unaccounted for. Upward and downward movements in the RSN were observed, and the maximum upward and downward distances were 40 cm and 100 cm, respectively. In general, the 15N-labeling method contributes to a deeper understanding of the fates of the RSN. Considering the low crop recovery of the RSN from deep soil layers, water and N saving practices should be adopted during crop production.

  4. Lucifer yellow filling of immunohistochemically pre-labeled neurons: a new method to characterize neuronal subpopulations.

    PubMed

    Galuske, R A; Delius, J A; Singer, W

    1993-07-01

    We describe a new technique for the morphological characterization of immunohistochemically labeled neuron populations. We demonstrate that it is possible to fill neurons iontophoretically with Lucifer Yellow (LY) in fixed slices of cat visual cortex after the respective cells have been identified by indirect immunofluorescence for the neural cell adhesion molecule N-CAM 180, with the VC1.1 antibody or with an antibody against glutamate dehydrogenase (GAD). Morphological analysis of the injected cells at the light and electron microscopic level revealed that the N-CAM 180-positive neurons share the features of neuropeptidergic cortical interneurons. Depending on the antibody applied, the immunohistochemical treatment had little or no noticeable effect on the quality of LY filling or on the preservation of morphological details of the pre-labeled cells. This makes the method described ideally suited for the light and electron microscopic examination of selected, immunologically characterized neuron subpopulations.

  5. N-(/sup 3/H)acetyl-labeling, a convenient method for radiolabeling of glycosaminoglycans

    SciTech Connect

    Hook, M.; Riesenfeld, J.; Lindahl, U.

    1982-01-15

    A method for the introduction of N-(/sup 3/H)acetyl groups into glycosaminoglycans is described. The procedure is based on (/sup 3/H)acetylation of N-unsubstituted hexosamine residues by treating the polysaccharides with (/sup 3/H)acetic anhydride. Preparations of heparin and heparin sulfate were found to contain significant numbers of N-unsubstituted hexosamine residues, as isolates. In contrast, such units could not be detected in chondroitin sulfate, dermatan sulfate, or hyaluronic acid. These polysaccharides were therefore subjected to partial N-deacetylation by reaction with hydrazine in the presence of hydrazine sulfate. After treatment with (/sup 3/H)acetic anhydride, the specific activities of the resulting labeled polysaccharide preparations ranged between 0.1 X 10/sup 6/ and 0.6 X 10/sup 6/ cpm /sup 3/H/..mu..g of uronic acid. The /sup 3/H-labeled polysaccharide preparations did not differ significantly from the corresponding unlabeled starting materials with regard to polyanion properties (chromatography on DEAE-cellulose) or polymer chain size (gel chromatography). Further, the radiolabeled polysaccharide derivatives were susceptible to specific enzymatic degradation (chondroitinase ABC and mammalian heparitinase) and retained their ability to interact specifically with certain proteins - for example, (/sup 3/H)heparin with antithrombin (/sup 3/H)hyaluronic acid oligosaccharides with chondroitin sulfate proteoglycan. These findings indicate that the labeling procedures did not induce any major structural derangement of the polysaccharide molecules. The method developed should be useful in providing labeled glycosaminoglycans for metabolic and enzymatic experiments as well as for studies on the interacion between glycosaminoglycans and other bilogical macromolecules.

  6. A multi-label learning based kernel automatic recommendation method for support vector machine.

    PubMed

    Zhang, Xueying; Song, Qinbao

    2015-01-01

    Choosing an appropriate kernel is very important and critical when classifying a new problem with Support Vector Machine. So far, more attention has been paid on constructing new kernels and choosing suitable parameter values for a specific kernel function, but less on kernel selection. Furthermore, most of current kernel selection methods focus on seeking a best kernel with the highest classification accuracy via cross-validation, they are time consuming and ignore the differences among the number of support vectors and the CPU time of SVM with different kernels. Considering the tradeoff between classification success ratio and CPU time, there may be multiple kernel functions performing equally well on the same classification problem. Aiming to automatically select those appropriate kernel functions for a given data set, we propose a multi-label learning based kernel recommendation method built on the data characteristics. For each data set, the meta-knowledge data base is first created by extracting the feature vector of data characteristics and identifying the corresponding applicable kernel set. Then the kernel recommendation model is constructed on the generated meta-knowledge data base with the multi-label classification method. Finally, the appropriate kernel functions are recommended to a new data set by the recommendation model according to the characteristics of the new data set. Extensive experiments over 132 UCI benchmark data sets, with five different types of data set characteristics, eleven typical kernels (Linear, Polynomial, Radial Basis Function, Sigmoidal function, Laplace, Multiquadric, Rational Quadratic, Spherical, Spline, Wave and Circular), and five multi-label classification methods demonstrate that, compared with the existing kernel selection methods and the most widely used RBF kernel function, SVM with the kernel function recommended by our proposed method achieved the highest classification performance.

  7. A Multi-Label Learning Based Kernel Automatic Recommendation Method for Support Vector Machine

    PubMed Central

    Zhang, Xueying; Song, Qinbao

    2015-01-01

    Choosing an appropriate kernel is very important and critical when classifying a new problem with Support Vector Machine. So far, more attention has been paid on constructing new kernels and choosing suitable parameter values for a specific kernel function, but less on kernel selection. Furthermore, most of current kernel selection methods focus on seeking a best kernel with the highest classification accuracy via cross-validation, they are time consuming and ignore the differences among the number of support vectors and the CPU time of SVM with different kernels. Considering the tradeoff between classification success ratio and CPU time, there may be multiple kernel functions performing equally well on the same classification problem. Aiming to automatically select those appropriate kernel functions for a given data set, we propose a multi-label learning based kernel recommendation method built on the data characteristics. For each data set, the meta-knowledge data base is first created by extracting the feature vector of data characteristics and identifying the corresponding applicable kernel set. Then the kernel recommendation model is constructed on the generated meta-knowledge data base with the multi-label classification method. Finally, the appropriate kernel functions are recommended to a new data set by the recommendation model according to the characteristics of the new data set. Extensive experiments over 132 UCI benchmark data sets, with five different types of data set characteristics, eleven typical kernels (Linear, Polynomial, Radial Basis Function, Sigmoidal function, Laplace, Multiquadric, Rational Quadratic, Spherical, Spline, Wave and Circular), and five multi-label classification methods demonstrate that, compared with the existing kernel selection methods and the most widely used RBF kernel function, SVM with the kernel function recommended by our proposed method achieved the highest classification performance. PMID:25893896

  8. Comparison and applications of label-free absolute proteome quantification methods on Escherichia coli.

    PubMed

    Arike, L; Valgepea, K; Peil, L; Nahku, R; Adamberg, K; Vilu, R

    2012-09-18

    Three different label-free proteome quantification methods--APEX, emPAI and iBAQ--were evaluated to measure proteome-wide protein concentrations in the cell. All the methods were applied to a sample from Escherichia coli chemostat culture. A Pearson squared correlation of approximately 0.6 among the three quantification methods was demonstrated. Importantly, the sum of quantified proteins by iBAQ and emPAI corresponded with the Lowry total protein quantification, demonstrating applicability of label-free methods for an accurate calculation of protein concentrations at the proteome level. The iBAQ method showed the best correlation between biological replicates, a normal distribution among all protein abundances, and the lowest variation among ribosomal protein abundances, which are expected to have equal amounts. Absolute quantitative proteome data enabled us to evaluate metabolic cost for protein synthesis and apparent catalytic activities of enzymes by integration with flux analysis. All the methods demonstrated similar ATP costs for protein synthesis for different cellular processes and that costs for expressing biomass synthesis related proteins were higher than those for energy generation. Importantly, catalytic activities of energy metabolism enzymes were an order or two higher than those of monomer synthesis. Interestingly, a staircase-like protein expression was demonstrated for most of the transcription units.

  9. Field measurement of seepage and evapotranspiration rate for a soil under plant cover: A comparison of soil water balance and tritium labeling procedure

    NASA Astrophysics Data System (ADS)

    Kreutzer, K.; Strebel, O.; Renger, M.

    1980-08-01

    Vertical water flux at 90 cm depth and evapotranspiration were measured in a loess Parabraunerde soil profile, under spring wheat and sugar beets, respectively, during a time period of nearly 21 months. Two field methods were compared: the HTO-tracer method (labeling soil water at a depth of 60 cm followed by core sampling) and the soil water balance method (measuring soil water suction and water content as a function of depth and time). Outside the vegetation season the results of the two methods agreed well, but not during the vegetation season. The reason is that the reference soil compartment, with its reference depth of 90 cm, lies within the root zone and the HTO-method does not correctly reflect the water flux through the roots and the water withdrawal by the roots from this reference compartment. It is shown, that after correcting the HTO-values for these root-activity-dependent effects, a good agreement between the two methods was found also during periods with root activity. Investigations with the HTO-method lead to inaccurate results if the reference depth or the median value of the tracer distribution lie within the zone of active roots.

  10. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... manufacturer. (iii) The manufacturer may include the ENERGY STAR logo on the bottom right corner of the label... may add the ENERGY STAR logo to labels on qualifying covered products; such manufacturers may add the ENERGY STAR logo to labels only on those covered products that are contemplated by the Memorandum...

  11. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... manufacturer. (iii) The manufacturer may include the ENERGY STAR logo on the bottom right corner of the label... may add the ENERGY STAR logo to labels on qualifying covered products; such manufacturers may add the ENERGY STAR logo to labels only on those covered products that are contemplated by the Memorandum...

  12. Stereo- and regioselective direct multi-deuterium-labeling methods for sugars.

    PubMed

    Sawama, Yoshinari; Yabe, Yuki; Iwata, Hiroki; Fujiwara, Yuta; Monguchi, Yasunari; Sajiki, Hironao

    2012-12-14

    Deuterium-labeled sugars can be utilized as powerful tools for the architectural analyses of high-sugar-containing molecules represented by the nucleic acids and glycoproteins, and chiral building blocks for the syntheses of new drug candidates (heavy drugs) due to their potential characteristics, such as simplifying the (1)H NMR spectra and the stability of C-D bonds compared with C-H bonds. We have established a direct and efficient synthetic method of deuterated sugars from non-labeled sugars by using the heterogeneous Ru/C-catalyzed H-D exchange reaction in D(2)O under a hydrogen atmosphere with perfect chemo- and stereoselectivities. The direct H-D exchange reaction can selectively proceed on carbons adjacent to the free hydroxyl groups, and the deuterium labeling of various pyranosides (such as glucose and disaccharides), as well as furanosides, represented by ribose and deoxyribose was realized. Furthermore, the desired number of deuterium atoms can be freely incorporated into selected positions by the site-selective protection of the hydroxyl groups using acetal-type protective groups because the deuterium exchange reaction never proceeds on positions adjacent to the protected hydroxyl groups. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Tracking Transplanted Stem Cells Using Magnetic Resonance Imaging and the Nanoparticle Labeling Method in Urology

    PubMed Central

    Kim, Jae Heon; Lee, Hong J.; Song, Yun Seob

    2015-01-01

    A reliable in vivo imaging method to localize transplanted cells and monitor their viability would enable a systematic investigation of cell therapy. Most stem cell transplantation studies have used immunohistological staining, which does not provide information about the migration of transplanted cells in vivo in the same host. Molecular imaging visualizes targeted cells in a living host, which enables determining the biological processes occurring in transplanted stem cells. Molecular imaging with labeled nanoparticles provides the opportunity to monitor transplanted cells noninvasively without sacrifice and to repeatedly evaluate them. Among several molecular imaging techniques, magnetic resonance imaging (MRI) provides high resolution and sensitivity of transplanted cells. MRI is a powerful noninvasive imaging modality with excellent image resolution for studying cellular dynamics. Several types of nanoparticles including superparamagnetic iron oxide nanoparticles and magnetic nanoparticles have been used to magnetically label stem cells and monitor viability by MRI in the urologic field. This review focuses on the current role and limitations of MRI with labeled nanoparticles for tracking transplanted stem cells in urology. PMID:26413510

  14. Tracking Transplanted Stem Cells Using Magnetic Resonance Imaging and the Nanoparticle Labeling Method in Urology.

    PubMed

    Kim, Jae Heon; Lee, Hong J; Song, Yun Seob

    2015-01-01

    A reliable in vivo imaging method to localize transplanted cells and monitor their viability would enable a systematic investigation of cell therapy. Most stem cell transplantation studies have used immunohistological staining, which does not provide information about the migration of transplanted cells in vivo in the same host. Molecular imaging visualizes targeted cells in a living host, which enables determining the biological processes occurring in transplanted stem cells. Molecular imaging with labeled nanoparticles provides the opportunity to monitor transplanted cells noninvasively without sacrifice and to repeatedly evaluate them. Among several molecular imaging techniques, magnetic resonance imaging (MRI) provides high resolution and sensitivity of transplanted cells. MRI is a powerful noninvasive imaging modality with excellent image resolution for studying cellular dynamics. Several types of nanoparticles including superparamagnetic iron oxide nanoparticles and magnetic nanoparticles have been used to magnetically label stem cells and monitor viability by MRI in the urologic field. This review focuses on the current role and limitations of MRI with labeled nanoparticles for tracking transplanted stem cells in urology.

  15. Some quantitative aspects of the labelling of proteins with 125I by the iodine monochloride method

    PubMed Central

    Ceska, M.; Sjödin, A. V.; Grossmüller, F.

    1971-01-01

    The labelling of proteins by the iodine monochloride method was studied by using a mathematical model. The equations used were primarily derived from the mass law equation of the isotopic exchange reaction between [125I]iodide and iodine monochloride. For convenient application, all equations were programmed into a computing desk-top calculator. To support the validity of the theoretical model, a series of iodinations of insulin were performed under various labelling conditions. The results of these experiments compare well with the theoretically derived values. Deviations from the theoretical values occurring at molar ratios of [125I]iodide to iodine monochloride < 0.1 and > 4.0 are explained and suggestions made about how to prevent them. The mathematical model was used to simulate the isotopic exchange, and the iodination reaction under various conditions, to study (a) the influence of the amount of [125I]iodide on the amount of [125I]iodine monochloride formed, (b) the influence of the specific radioactivity of [125I]iodide on the amount of [125I]iodine monochloride formed, and (c) the influence of the specific radioactivity of [125I]iodide on the number of millicuries needed for labelling to a desired extent. PMID:5116527

  16. A COMPARISON OF METHODS FOR TEACHING RECEPTIVE LABELING TO CHILDREN WITH AUTISM SPECTRUM DISORDERS

    PubMed Central

    Grow, Laura L; Carr, James E; Kodak, Tiffany M; Jostad, Candice M; Kisamore, April N

    2011-01-01

    Many early intervention curricular manuals recommend teaching auditory-visual conditional discriminations (i.e., receptive labeling) using the simple-conditional method in which component simple discriminations are taught in isolation and in the presence of a distracter stimulus before the learner is required to respond conditionally. Some have argued that this procedure might be susceptible to faulty stimulus control such as stimulus overselectivity (Green, 2001). Consequently, there has been a call for the use of alternative teaching procedures such as the conditional-only method, which involves conditional discrimination training from the onset of intervention. The purpose of the present study was to compare the simple-conditional and conditional-only methods for teaching receptive labeling to 3 young children diagnosed with autism spectrum disorders. The data indicated that the conditional-only method was a more reliable and efficient teaching procedure. In addition, several error patterns emerged during training using the simple-conditional method. The implications of the results with respect to current teaching practices in early intervention programs are discussed. PMID:21941380

  17. A comparison of methods for teaching receptive labeling to children with autism spectrum disorders.

    PubMed

    Grow, Laura L; Carr, James E; Kodak, Tiffany M; Jostad, Candice M; Kisamore, April N

    2011-01-01

    Many early intervention curricular manuals recommend teaching auditory-visual conditional discriminations (i.e., receptive labeling) using the simple-conditional method in which component simple discriminations are taught in isolation and in the presence of a distracter stimulus before the learner is required to respond conditionally. Some have argued that this procedure might be susceptible to faulty stimulus control such as stimulus overselectivity (Green, 2001). Consequently, there has been a call for the use of alternative teaching procedures such as the conditional-only method, which involves conditional discrimination training from the onset of intervention. The purpose of the present study was to compare the simple-conditional and conditional-only methods for teaching receptive labeling to 3 young children diagnosed with autism spectrum disorders. The data indicated that the conditional-only method was a more reliable and efficient teaching procedure. In addition, several error patterns emerged during training using the simple-conditional method. The implications of the results with respect to current teaching practices in early intervention programs are discussed.

  18. Environmental labeling of car tires--toxicity to Daphnia magna can be used as a screening method.

    PubMed

    Wik, Anna; Dave, Göran

    2005-02-01

    Car tires contain several water-soluble compounds that can leach into water and have toxic effects on aquatic organisms. Due to tire wear, 10,000 tonnes of rubber particles end up along the Swedish roads every year. This leads to a diffuse input of emissions of several compounds. Emissions of polyaromatic hydrocarbons (PAHs) are of particular concern. PAHs are ingredients of the high aromatic oil (HA oil) that is used in the rubber as a softener and as a filler. The exclusion of HA oils from car tires has started, and an environmental labeling of tires could make HA oils obsolete. The toxicity to Daphnia magna from 12 randomly selected car tires was tested in this study. Rubber from the tread of the tires was grated into small pieces, to simulate material from tire wear, and the rubber was equilibrated with dilution water for 72 h before addition of test organisms. The 24-h EC50s of the rubber pieces ranged from 0.29 to 32 gl-1, and the 48-h EC50s ranged from 0.0625 to 2.41 gl-1. Summer tires were more toxic than winter tires. After the 48-h exposure, the daphnids were exposed to UV-light for 2 h, to determine if the tires contained compounds that were phototoxic. After UV-activation the EC50s ranged from 0.0625 to 0.38 gl-1. Four of the 12 tires had a very distinct photoactivation, with a toxicity increase of >10 times. This study has shown that the used method for toxicity testing with Daphnia magna according to ISO 6341 could be used as a basis for environmental labeling of car tires.

  19. Family-Joining: A Fast Distance-Based Method for Constructing Generally Labeled Trees

    PubMed Central

    Kalaghatgi, Prabhav; Pfeifer, Nico; Lengauer, Thomas

    2016-01-01

    The widely used model for evolutionary relationships is a bifurcating tree with all taxa/observations placed at the leaves. This is not appropriate if the taxa have been densely sampled across evolutionary time and may be in a direct ancestral relationship, or if there is not enough information to fully resolve all the branching points in the evolutionary tree. In this article, we present a fast distance-based agglomeration method called family-joining (FJ) for constructing so-called generally labeled trees in which taxa may be placed at internal vertices and the tree may contain polytomies. FJ constructs such trees on the basis of pairwise distances and a distance threshold. We tested three methods for threshold selection, FJ-AIC, FJ-BIC, and FJ-CV, which minimize Akaike information criterion, Bayesian information criterion, and cross-validation error, respectively. When compared with related methods on simulated data, FJ-BIC was among the best at reconstructing the correct tree across a wide range of simulation scenarios. FJ-BIC was applied to HIV sequences sampled from individuals involved in a known transmission chain. The FJ-BIC tree was found to be compatible with almost all transmission events. On average, internal branches in the FJ-BIC tree have higher bootstrap support than branches in the leaf-labeled bifurcating tree constructed using RAxML. 36% and 25% of the internal branches in the FJ-BIC tree and RAxML tree, respectively, have bootstrap support greater than 70%. To the best of our knowledge the method presented here is the first attempt at modeling evolutionary relationships using generally labeled trees. PMID:27436007

  20. Family-Joining: A Fast Distance-Based Method for Constructing Generally Labeled Trees.

    PubMed

    Kalaghatgi, Prabhav; Pfeifer, Nico; Lengauer, Thomas

    2016-10-01

    The widely used model for evolutionary relationships is a bifurcating tree with all taxa/observations placed at the leaves. This is not appropriate if the taxa have been densely sampled across evolutionary time and may be in a direct ancestral relationship, or if there is not enough information to fully resolve all the branching points in the evolutionary tree. In this article, we present a fast distance-based agglomeration method called family-joining (FJ) for constructing so-called generally labeled trees in which taxa may be placed at internal vertices and the tree may contain polytomies. FJ constructs such trees on the basis of pairwise distances and a distance threshold. We tested three methods for threshold selection, FJ-AIC, FJ-BIC, and FJ-CV, which minimize Akaike information criterion, Bayesian information criterion, and cross-validation error, respectively. When compared with related methods on simulated data, FJ-BIC was among the best at reconstructing the correct tree across a wide range of simulation scenarios. FJ-BIC was applied to HIV sequences sampled from individuals involved in a known transmission chain. The FJ-BIC tree was found to be compatible with almost all transmission events. On average, internal branches in the FJ-BIC tree have higher bootstrap support than branches in the leaf-labeled bifurcating tree constructed using RAxML. 36% and 25% of the internal branches in the FJ-BIC tree and RAxML tree, respectively, have bootstrap support greater than 70%. To the best of our knowledge the method presented here is the first attempt at modeling evolutionary relationships using generally labeled trees.

  1. Assessing of distribution, mobility and bioavailability of exogenous Pb in agricultural soils using isotopic labeling method coupled with BCR approach.

    PubMed

    Huang, Zhi-Yong; Xie, Hong; Cao, Ying-Lan; Cai, Chao; Zhang, Zhi

    2014-02-15

    The contamination of Pb in agricultural soils is one of the most important ecological problems, which potentially results in serious health risk on human health through food chain. Hence, the fate of exogenous Pb contaminated in agricultural soils is needed to be deeply explored. By spiking soils with the stable enriched isotopes of (206)Pb, the contamination of exogenous Pb(2+) ions in three agricultural soils sampled from the estuary areas of Jiulong River, China was simulated in the present study, and the distribution, mobility and bioavailability of exogenous Pb in the soils were investigated using the isotopic labeling method coupled with a four-stage BCR (European Community Bureau of Reference) sequential extraction procedure. Results showed that about 60-85% of exogenous Pb was found to distribute in reducible fractions, while the exogenous Pb in acid-extractable fractions was less than 1.0%. After planting, the amounts of exogenous Pb presenting in acid-extractable, reducible and oxidizable fractions in rhizospheric soils decreased by 60-66%, in which partial exogenous Pb was assimilated by plants while most of the metal might transfer downward due to daily watering and applying fertilizer. The results show that the isotopic labeling technique coupled with sequential extraction procedures enables us to explore the distribution, mobility and bioavailability of exogenous Pb contaminated in soils, which may be useful for the further soil remediation. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Total energy expenditure in burned children using the doubly labeled water technique

    SciTech Connect

    Goran, M.I.; Peters, E.J.; Herndon, D.N.; Wolfe, R.R. )

    1990-10-01

    Total energy expenditure (TEE) was measured in 15 burned children with the doubly labeled water technique. Application of the technique in burned children required evaluation of potential errors resulting from nutritional intake altering background enrichments during studies and from the high rate of water turnover relative to CO2 production. Five studies were discarded because of these potential problems. TEE was 1.33 +/- 0.27 times predicted basal energy expenditure (BEE), and in studies where resting energy expenditure (REE) was simultaneously measured, TEE was 1.18 +/- 0.17 times REE, which in turn was 1.16 +/- 0.10 times predicted BEE. TEE was significantly correlated with measured REE (r2 = 0.92) but not with predicted BEE. These studies substantiate the advantage of measuring REE to predict TEE in severely burned patients as opposed to relying on standardized equations. Therefore we recommend that optimal nutritional support will be achieved in convalescent burned children by multiplying REE by an activity factor of 1.2.

  3. Microwave accelerated labeling methods in the synthesis of radioligands for positron emission tomography imaging.

    PubMed

    Kallmerten, Amy E; Alexander, Abigail; Wager, Krista M; Jones, Graham B

    2011-10-01

    Nuclear imaging using positron emission tomography [PET] is a powerful technique with clinical applications which include oncology, cardiovascular disease and CNS disorders. Conventional chemical syntheses of the short half-life radionuclides used in the process however imposes numerous limitations on scope of available ligands. By utilizing microwave assisted synthesis methods many of these limitations can be overcome, paving the way for the design of diverse families of agents with defined cellular targets. This review will survey recent developments in the field with emphasis on the period 2006-2011. Positron emission tomography [PET] has become one of the most powerful in vivo imaging modalities, capable of delivering mm3 resolution of radiotracer distribution and metabolism [1]. When combined with anatomic imaging methods (MRI, CT) co-registered multimode images offer the potential to track metabolic and physiologic events in diseased states and guide and accelerate clinical trials of investigational new drugs. Also, this same methodology can be used to evaluate first pass pharmacokinetics/pharmacodynamics in early stage drug discovery. Though powerful as a technique only a limited number of drugs have seen clinical use and to date only one drug 2-fluoro-deoxy-D-glucose (FDG) has received FDA approval [2]. One of the drawbacks of PET imaging is the need for tracers labeled with an appropriate nuclide and the half-lives of these agents places special constraints on the chemical synthesis. Among the most popular are 11C (t½ =20.4 min) and 18F (t ½ =109.8 min) labeled compounds and this has resulted in a resurgence of interest in practical application of their chemistries [3,4]. This review will focus on microwave mediated methods of acceleration of organic reactions used for the production of labeled PET image contrast agents, with emphasis on the five year period 2006 to 2011.

  4. A mild and reliable method to label enveloped virus with quantum dots by copper-free click chemistry.

    PubMed

    Hao, Jian; Huang, Li-Li; Zhang, Rui; Wang, Han-Zhong; Xie, Hai-Yan

    2012-10-02

    Real-time tracking of the dynamic process of virus invasion is crucial to understanding the infection mechanism. For successful tracking, efficient labeling methods are indispensable. In this paper, we report a mild and reliable method for labeling viruses, especially with regard to easily disabled enveloped viruses. The copper-free click chemistry has been used to label enveloped viruses with quantum dots (QDs) by linking virions modified with azide to the QDs derived with dibenzocyclooctynes (DBCO). Both vaccinia virus (VACV) and avian influenza A virus (H9N2) can be specifically and rapidly labeled under mild conditions, with a labeling efficiency of more than 80%. The labeled virions were of intact infectivity, and their fluorescence was strong enough to realize single-virion tracking. Compared to previously reported methods, our method is less destructive, reliable, and universal, without specific requirements for the type and structure of viruses to be labeled, which has laid the foundation for long-term dynamic visualization of virus infection process.

  5. Enrichment of H(2)(17)O from tap water, characterization of the enriched water, and properties of several (17)O-labeled compounds.

    PubMed

    Prasad, Brinda; Lewis, Andrew R; Plettner, Erika

    2011-01-01

    A low-abundance form of water, H(2)(17)O, was enriched from 0.04% to ∼90% by slow evaporation and fractional distillation of tap water. The density and refractive index for H(2)(17)O are reported. Gas chromatography-mass spectrometry (GC-MS) of (16)O- and (17)O-1-hexanols and their trimethyl silyl ethers and of (16)O- and (17)O-hexamethyl disiloxanes was used to determine the percentage of (17)O enrichment in the H(2)(17)O. Furthermore, the chemical shifts of labeled and nonlabeled water dissolved in CDCl(3) differed sufficiently that we could verify the enrichment of H(2)(17)O. (17)O hexanol was synthesized by the reaction of iodohexane with Na(17)OH. (17)O-Labeled trimethylsilanol and (17)O-labeled hexamethyldisiloxane were prepared by the reaction of H(2)(17)O with bis(trimethylsilyl)trifluoroacetamide (BSTFA). To generate standards for (17)O NMR, H(2)(17)O(2), and (17)O camphor were prepared. H(2)(17)O was electrolyzed to form (17)O-labeled hydrogen peroxide which was quantified using two colorimetric assays. (17)O-Labeled camphor was prepared by exchanging the ketone oxygen of camphor using H(2)(17)O. The (17)O-labeled compounds were characterized using (17)O, (1)H, and (13)C NMR and GC-MS. While we were characterizing the labeled camphor, we also detected an unexpected oxygen exchange reaction of primary alcohols, catalyzed by electrophilic ketones such as camphor. The reaction is a displacement of the alcohol OH group by water. This is an example of the usefulness of (17)O NMR in the study of a reaction mechanism that has not been noticed previously.

  6. A coding method for efficient subgraph querying on vertex- and edge-labeled graphs.

    PubMed

    Zhu, Lei; Song, Qinbao; Guo, Yuchen; Du, Lei; Zhu, Xiaoyan; Wang, Guangtao

    2014-01-01

    Labeled graphs are widely used to model complex data in many domains, so subgraph querying has been attracting more and more attention from researchers around the world. Unfortunately, subgraph querying is very time consuming since it involves subgraph isomorphism testing that is known to be an NP-complete problem. In this paper, we propose a novel coding method for subgraph querying that is based on Laplacian spectrum and the number of walks. Our method follows the filtering-and-verification framework and works well on graph databases with frequent updates. We also propose novel two-step filtering conditions that can filter out most false positives and prove that the two-step filtering conditions satisfy the no-false-negative requirement (no dismissal in answers). Extensive experiments on both real and synthetic graphs show that, compared with six existing counterpart methods, our method can effectively improve the efficiency of subgraph querying.

  7. Intein applications: from protein purification and labeling to metabolic control methods.

    PubMed

    Wood, David W; Camarero, Julio A

    2014-05-23

    The discovery of inteins in the early 1990s opened the door to a wide variety of new technologies. Early engineered inteins from various sources allowed the development of self-cleaving affinity tags and new methods for joining protein segments through expressed protein ligation. Some applications were developed around native and engineered split inteins, which allow protein segments expressed separately to be spliced together in vitro. More recently, these early applications have been expanded and optimized through the discovery of highly efficient trans-splicing and trans-cleaving inteins. These new inteins have enabled a wide variety of applications in metabolic engineering, protein labeling, biomaterials construction, protein cyclization, and protein purification.

  8. .sup.123m Te-Labeled biochemicals and method of preparation

    DOEpatents

    Knapp, Jr., Furn F.

    1980-01-01

    A novel class of .sup.123m Te-labeled steroids and amino acids is provided by the method of reacting a .sup.123m Te symmetric diorgano ditelluride with a hydride reducing agent and a source of alkali metal ions to form an alkali metal organo telluride. The alkali metal organo telluride is reacted with a primary halogenated steroidal side chain, amino acid, or amino acid precursor such as hydantoin. The novel compounds are useful as biological tracers and as organal imaging agents.

  9. Automatic brain extraction methods for T1 magnetic resonance images using region labeling and morphological operations.

    PubMed

    Somasundaram, K; Kalaiselvi, T

    2011-08-01

    In this work we propose two brain extraction methods (BEM) that solely depend on the brain anatomy and its intensity characteristics. Our methods are simple, unsupervised and knowledge based. Using an adaptive intensity thresholding method on the magnetic resonance images of head scans, a binary image is obtained. The binary image is labeled using the anatomical facts that the scalp is the boundary between head and background, and the skull is the boundary separating brain and scalp. A run length scheme is applied on the labeled image to get a rough brain mask. Morphological operations are then performed to obtain the fine brain on the assumption that brain is the largest connected component (LCC). But the LCC concept failed to work on some slices where brain is composed of more than one connected component. To solve this problem a 3-D approach is introduced in the BEM. Experimental results on 61 sets of T1 scans taken from MRI scan center and neuroimage web services showed that our methods give better results than the popular methods, FSL's Brain Extraction Tool (BET), BrainSuite's Brain Surface Extractor (BSE) gives results comparable to that of Model-based Level Sets (MLS) and works well even where MLS failed. The average Dice similarity index computed using the "Gold standard" and the specificity values are 0.938 and 0.992, respectively, which are higher than that for BET, BSE and MLS. The average processing time by one of our methods is ≈1s/slice, which is smaller than for MLS, which is ≈4s/slice. One of our methods produces the lowest false positive rate of 0.075, which is smaller than that for BSE, BET and MLS. It is independent of imaging orientation and works well for slices with abnormal features like tumor and lesion in which the existing methods fail in certain cases.

  10. Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests

    PubMed Central

    Laitinen, Mika PA; Salmela, Jari; Gilbert, Leona; Kaivola, Risto; Tikkala, Topi; Oker-Blom, Christian; Pekola, Jukka; Vuento, Matti

    2009-01-01

    A rapid method and instrumentation for quantification of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The first anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the first anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane. The amount of superparamagnetic particle-labelled monoclonal anti-hCG bound to the detection zone was directly proportional to the amount of hCG present in the sample as detected by measuring magnetization in the detector coil. The method had a practical detection limit of 20 U/l (54 nM) of hCG per 5 μl of human urine and a linear range of three decades from 20 U/l to 10 000 U/l. In addition, the analysis was completed within less than 10 minutes. Thus, the test format should be suitable for fast detection and monitoring of a large variety of clinically important parameters and analytes. PMID:24198463

  11. Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests.

    PubMed

    Laitinen, Mika Pa; Salmela, Jari; Gilbert, Leona; Kaivola, Risto; Tikkala, Topi; Oker-Blom, Christian; Pekola, Jukka; Vuento, Matti

    2009-01-01

    A rapid method and instrumentation for quantification of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The first anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the first anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane. The amount of superparamagnetic particle-labelled monoclonal anti-hCG bound to the detection zone was directly proportional to the amount of hCG present in the sample as detected by measuring magnetization in the detector coil. The method had a practical detection limit of 20 U/l (54 nM) of hCG per 5 μl of human urine and a linear range of three decades from 20 U/l to 10 000 U/l. In addition, the analysis was completed within less than 10 minutes. Thus, the test format should be suitable for fast detection and monitoring of a large variety of clinically important parameters and analytes.

  12. Reported Energy Intake Accuracy Compared to Doubly Labeled Water and Usability of the Mobile Food Record among Community Dwelling Adults.

    PubMed

    Boushey, Carol J; Spoden, Melissa; Delp, Edward J; Zhu, Fengqing; Bosch, Marc; Ahmad, Ziad; Shvetsov, Yurii B; DeLany, James P; Kerr, Deborah A

    2017-03-22

    The mobile Food Record (mFR) is an image-based dietary assessment method for mobile devices. The study primary aim was to test the accuracy of the mFR by comparing reported energy intake (rEI) to total energy expenditure (TEE) using the doubly labeled water (DLW) method. Usability of the mFR was assessed by questionnaires before and after the study. Participants were 45 community dwelling men and women, 21-65 years. They were provided pack-out meals and snacks and encouraged to supplement with usual foods and beverages not provided. After being dosed with DLW, participants were instructed to record all eating occasions over a 7.5 days period using the mFR. Three trained analysts estimated rEI from the images sent to a secure server. rEI and TEE correlated significantly (Spearman correlation coefficient of 0.58, p < 0.0001). The mean percentage of underreporting below the lower 95% confidence interval of the ratio of rEI to TEE was 12% for men (standard deviation (SD) ± 11%) and 10% for women (SD ± 10%). The results demonstrate the accuracy of the mFR is comparable to traditional dietary records and other image-based methods. No systematic biases could be found. The mFR was received well by the participants and usability was rated as easy.

  13. Reported Energy Intake Accuracy Compared to Doubly Labeled Water and Usability of the Mobile Food Record among Community Dwelling Adults

    PubMed Central

    Boushey, Carol J.; Spoden, Melissa; Delp, Edward J.; Zhu, Fengqing; Bosch, Marc; Ahmad, Ziad; Shvetsov, Yurii B.; DeLany, James P.; Kerr, Deborah A.

    2017-01-01

    The mobile Food Record (mFR) is an image-based dietary assessment method for mobile devices. The study primary aim was to test the accuracy of the mFR by comparing reported energy intake (rEI) to total energy expenditure (TEE) using the doubly labeled water (DLW) method. Usability of the mFR was assessed by questionnaires before and after the study. Participants were 45 community dwelling men and women, 21–65 years. They were provided pack-out meals and snacks and encouraged to supplement with usual foods and beverages not provided. After being dosed with DLW, participants were instructed to record all eating occasions over a 7.5 days period using the mFR. Three trained analysts estimated rEI from the images sent to a secure server. rEI and TEE correlated significantly (Spearman correlation coefficient of 0.58, p < 0.0001). The mean percentage of underreporting below the lower 95% confidence interval of the ratio of rEI to TEE was 12% for men (standard deviation (SD) ± 11%) and 10% for women (SD ± 10%). The results demonstrate the accuracy of the mFR is comparable to traditional dietary records and other image-based methods. No systematic biases could be found. The mFR was received well by the participants and usability was rated as easy. PMID:28327502

  14. A deuterium-based labeling technique for the investigation of rooting depths, water uptake dynamics and unsaturated zone water transport in semiarid environments

    NASA Astrophysics Data System (ADS)

    Beyer, M.; Koeniger, P.; Gaj, M.; Hamutoko, J. T.; Wanke, H.; Himmelsbach, T.

    2016-02-01

    Non- or minimum-invasive methods for the quantification of rooting depths of plants are rare, in particular in (semi-)arid regions; yet, this information is crucial for the parameterization of SVAT (Soil-Vegetation-Atmosphere Transfer) models and understanding of processes within the hydrological cycle. We present a technique utilizing the stable isotope deuterium (2H) applied as artificial tracer to investigate the vertical extent of the root zone, characterize water uptake dynamics of trees and shrubs at different depths and monitor transport of water through the unsaturated zone of dry environments. One liter of 35% deuterated water (2H2O) was punctually applied at several depths (0.5 m, 1 m, 2 m, 2.5 m and 4 m) at six different plots at a natural forested site in the Cuvelai-Etosha Basin (CEB), Namibia/Angola. Subsequently, uptake of the tracer was monitored by collecting plant samples (xylem and transpired water) up to seven days after tracer injection. Soil profiles at the plots were taken after the campaign and again after six months in order to evaluate the transport and distribution of 2H within the unsaturated zone. Of 162 plant samples taken, 31 samples showed clear signals of artificially introduced 2H, of which all originate from the plots labeled up to 2 m depth. No artificially injected 2H was found in plants when tracer application occurred deeper than 2 m. Results further indicate a sharing of water resources between the investigated shrubs and trees in the upper 1 m whilst tree roots seem to have better access to deeper layers of the unsaturated zone. The soil profiles taken after six months reveal elevated 2H-concentrations from depths as great as 4 m up to 1 m below surface indicating upward transport of water vapor. Purely diffuse transport towards the soil surface yielded an estimated 0.4 mm over the dry season. Results are of particular significance for a more precise parameterization of SVAT models and the formulation of water balances in

  15. Facile strain analysis of largely bending films by a surface-labelled grating method

    NASA Astrophysics Data System (ADS)

    Akamatsu, Norihisa; Tashiro, Wataru; Saito, Keisuke; Mamiya, Jun-Ichi; Kinoshita, Motoi; Ikeda, Tomiki; Takeya, Jun; Fujikawa, Shigenori; Priimagi, Arri; Shishido, Atsushi

    2014-06-01

    Mechanical properties of flexible films, for example surface strain of largely bending films, are key to design of stretchable electronic devices, wearable biointegrated devices, and soft microactuators/robots. However, existing methods are mainly based on strain-gauge measurements that require miniaturized array sensors, lead wires, and complicated calibrations. Here we introduce a facile method, based on surface-labelled gratings, for two-dimensional evaluation of surface strains in largely bending films. With this technique, we demonstrate that soft-matter mechanics can be distinct from the mechanics of hard materials. In particular, liquid-crystalline elastomers may undergo unconventional bending in three dimensions, in which both the inner and outer surfaces of the bending film are compressed. We also show that this method can be applied to amorphous elastomeric films, which highlights the general importance of this new mechanical evaluation tool in designing soft-matter-based electronic/photonic as well as biointegrated materials.

  16. Facile strain analysis of largely bending films by a surface-labelled grating method

    PubMed Central

    Akamatsu, Norihisa; Tashiro, Wataru; Saito, Keisuke; Mamiya, Jun-ichi; Kinoshita, Motoi; Ikeda, Tomiki; Takeya, Jun; Fujikawa, Shigenori; Priimagi, Arri; Shishido, Atsushi

    2014-01-01

    Mechanical properties of flexible films, for example surface strain of largely bending films, are key to design of stretchable electronic devices, wearable biointegrated devices, and soft microactuators/robots. However, existing methods are mainly based on strain-gauge measurements that require miniaturized array sensors, lead wires, and complicated calibrations. Here we introduce a facile method, based on surface-labelled gratings, for two-dimensional evaluation of surface strains in largely bending films. With this technique, we demonstrate that soft-matter mechanics can be distinct from the mechanics of hard materials. In particular, liquid-crystalline elastomers may undergo unconventional bending in three dimensions, in which both the inner and outer surfaces of the bending film are compressed. We also show that this method can be applied to amorphous elastomeric films, which highlights the general importance of this new mechanical evaluation tool in designing soft-matter-based electronic/photonic as well as biointegrated materials. PMID:24948462

  17. Humidity-conditioned gravimetric method to measure the water content of hydrogel contact lens materials.

    PubMed

    Galas, S L; Enns, J B

    1993-07-01

    A method to determine the humidity-conditioned gravimetric water content of hydrogel contact lens materials has been developed, in which errors due to blotting have been eliminated by conditioning the lens in a series of relative humidity (RH) environments before measuring the water content gravimetrically, and then extrapolating the water content to 100% RH. This method has been used to determine the water contents of representative materials from each of the four FDA lens groups, which were compared with their labeled values, as well as with values obtained from refractive index measurements. The deviation of the water content of soft contact lenses as measured by refractive index from that obtained gravimetrically increased as the water content decreased. The humidity-conditioned gravimetric method to determine water content of hydrophilic contact lenses is being proposed as an International Organization for Standardization (ISO) standard, as an improvement over the gravimetric and refractive index methods.

  18. Development of deuterium labeling method based on the heterogeneous platinum group metal-catalyzed C-H activation.

    PubMed

    Sajiki, Hironao

    2013-01-01

    Deuterium (D) labeled compounds are utilized in various scientific fields such as mechanistic elucidation of reactions, preparation of new functional materials, tracers for microanalysis, deuterium labeled heavy drugs and so on. Although the H-D exchange reaction is a straightforward method to produce deuterated organic compounds, many precedent methods require expensive deuterium gas and/or harsh reaction conditions. A part of our leading research agendas is intended to the development of novel and functional heterogeneous platinum-group catalysts and the reclamation of unknown functionalities of existing heterogeneous platinum-group catalysts. During the course of the study, benzylic positions of substrates were site-selectively deuterated under mild and palladium-on-carbon (Pd/C)-catalyzed hydrogenation conditions in heavy water (D2O). Heat conditions promoted the H-D exchange reactivity and facilitated the H-D exchange reaction at not only the benzylic sites but also inactive C-H bonds and heterocyclic nuclei. It is noteworthy that platinum-on-carbon (Pt/C) indicated a quite high affinity toward aromatic nuclei, and the H-D exchange reaction was strongly enhanced by the use of Pt/C as a catalyst under milder conditions. The mixed use of Pd/C and Pt/C was found to be more efficient in the H-D exchange reaction compared to the independent use of Pd/C or Pt/C. Furthermore, simple alkanes could also be efficiently deuterated under rhodium-on-carbon (Rh/C)-catalyzed conditions. The use of ruthenium-on-carbon (Ru/C) enabled the regiospecific and efficient deuterium incorporation at α-positions of alcohols and results were applied as a regio- and stereoselective multi-deuteration method of sugar derivatives.

  19. Improved method for the direct labeling of antibodies with Tc-99m

    SciTech Connect

    Rhodes, B.A.; Hawkins, E.; Budd, P. ); Deleide, G.; Seccamani, E.; Bonino, C. )

    1990-01-01

    Antibodies and antibody fragments have been treated with stannous chloride or organic reducing agents to reduce disulfide bonds, providing sulfhydryl groups for binding reduced Tc-99m. The reduced antibody, additives and stannous salts are lyophilized. To radiolabel, sodium pertechnetate solution is added, which dissolves the protein and other reagents. The pertechnetate is reduced by the stannous ions and becomes bound to the antibody. After radiolabeling the shelf-life of the product exceeds the half-life of the radionuclide. One of the more effective additives is human serum albumin, which serves as a carrier protein, agent to protect against autoradiolysis and possibly as a transfer ligand. Many different antibodies have been labeled using this method. The most widely studied antibody is an anti-melanoma fragment which has now been used clinically in more than 1000 cases and has been proven effective for the diagnostic localization of melanoma. In summary, a single vial, one step procedure for the direct labeling of antibodies in the presence of human serum albumin has been developed, extensively characterized, and clinically validated. The method is used with murine monoclonal IgG fragments, IgM and human gamma globulin. 4 refs., 10 figs.

  20. Indirect MRI of (17) o-labeled water using steady-state sequences: Signal simulation and preclinical experiment.

    PubMed

    Kudo, Kohsuke; Harada, Taisuke; Kameda, Hiroyuki; Uwano, Ikuko; Yamashita, Fumio; Higuchi, Satomi; Yoshioka, Kunihiro; Sasaki, Makoto

    2017-08-31

    Few studies have been reported for T2 -weighted indirect (17) O imaging. To evaluate the feasibility of steady-state sequences for indirect (17) O brain imaging. Signal simulation, phantom measurements, and prospective animal experiments were performed in accordance with the institutional guidelines for animal experiments. Signal simulations of balanced steady-state free precession (bSSFP) were performed for concentrations of (17) O ranging from 0.037-1.600%. Phantom measurements with concentrations of (17) O water ranging from 0.037-1.566% were also conducted. Six healthy beagle dogs were scanned with intravenous administration of 20% (17) O-labeled water (1 mL/kg). Dynamic 3D-bSSFP scans were performed at 3T MRI. (17) O-labeled water was injected 60 seconds after the scan start, and the total scan duration was 5 minutes. Based on the result of signal simulation and phantom measurement, signal changes in the beagle dogs were measured and converted into (17) O concentrations. The (17) O concentrations were averaged for every 15 seconds, and compared to the baseline (30-45 sec) with Dunnett's multiple comparison tests. Signal simulation revealed that the relationships between (17) O concentration and the natural logarithm of relative signals were linear. The intraclass correlation coefficient between relative signals in phantom measurement and signal simulations was 0.974. In the animal experiments, significant increases in (17) O concentration (P < 0.05) were observed 60 seconds after the injection of (17) O. At the end of scanning, mean respective (17) O concentrations of 0.084 ± 0.026%, 0.117 ± 0.038, 0.082 ± 0.037%, and 0.049 ± 0.004% were noted for the cerebral cortex, cerebellar cortex, cerebral white matter, and ventricle. Dynamic steady-state sequences were feasible for indirect (17) O imaging, and absolute quantification was possible. This method can be applied for the measurement of permeability and blood flow in the brain, and for

  1. Determinants of the energy costs of light activities: inferences for interpreting doubly labeled water data.

    PubMed

    Schoeller, D A; Jefford, G

    2002-01-01

    To investigate the controversy regarding the means of adjusting the energy cost of physical activity measured by doubly labeled water for differences in body size. We performed a cross-sectional study of the energy costs of carefully reproduced light activities with careful control for fidgeting and other unnecessary movement. :The study was performed in 23 healthy, young to middle aged adults with body masses between 39 and 118 kg. Energy expenditure was measured by respiratory gas exchange while subjects performed controlled light activities representative of activities of daily life. Body composition was measured by bioelectrical impedance analysis. The energy costs of individual controlled light activities were proportional to body weight except for mock vacuum cleaning. When the energy costs of all five activities were taken together, allometric regression analysis indicated that the energy cost of these light activities was proportional to body weight (slope=0.88+/-0.07), but not to fat-free mass (1.24+/-0.10), fat mass (0.27+/-0.03) or resting metabolic rate (1.43+/-0.12). Normalization of energy expenditure of physical activity by division by body weight is an appropriate means for comparing the volume (intensity x time) of physical activity between individuals of different body size.

  2. (Bio)degradation of glyphosate in water-sediment microcosms - A stable isotope co-labeling approach.

    PubMed

    Wang, Shizong; Seiwert, Bettina; Kästner, Matthias; Miltner, Anja; Schäffer, Andreas; Reemtsma, Thorsten; Yang, Qi; Nowak, Karolina M

    2016-08-01

    Glyphosate and its metabolite aminomethylphosphonic acid (AMPA) are frequently detected in water and sediments. Up to date, there are no comprehensive studies on the fate of glyphosate in water-sediment microcosms according to OECD 308 guideline. Stable isotope co-labeled (13)C3(15)N-glyphosate was used to determine the turnover mass balance, formation of metabolites, and formation of residues over a period of 80 days. In the water-sediment system, 56% of the initial (13)C3-glyphosate equivalents was ultimately mineralized, whereas the mineralization in the water system (without sediment) was low, reaching only 2% of (13)C-glyphosate equivalents. This finding demonstrates the key role of sediments in its degradation. Glyphosate was detected below detection limit in the water compartment on day 40, but could still be detected in the sediments, ultimately reaching 5% of (13)C3(15)N-glyphosate equivalents. A rapid increase in (13)C(15)N-AMPA was noted after 10 days, and these transformation products ultimately constituted 26% of the (13)C3-glyphosate equivalents and 79% of the (15)N-glyphosate equivalents. In total, 10% of the (13)C label and 12% of the (15)N label were incorporated into amino acids, indicating no risk bearing biogenic residue formation from (13)C3(15)N-glyphosate. Initially, glyphosate was biodegraded via the sarcosine pathway related to microbial growth, as shown by co-labeled (13)C(15)N-glycine and biogenic residue formation. Later, degradation via AMPA dominated under starvation conditions, as shown by the contents of (13)C-glycine. The presented data provide the first evidence of the speciation of the non-extractable residues as well as the utilization of glyphosate as a carbon and nitrogen source in the water-sediment system. This study also highlights the contribution of both the sarcosine and the AMPA degradation pathways under these conditions.

  3. An automatic labeling bifurcation method for intracoronary optical coherence tomography images

    NASA Astrophysics Data System (ADS)

    Macedo, Maysa M. G.; Takimura, Celso K.; Lemos, Pedro A.; Gutierrez, Marco A.

    2015-03-01

    Vessel branchings are critical vascular locations from the clinical point of view. In these sites, the arterial hemodynamic plays a relevant role in the progression of atherosclerosis, an important vascular pathology. In this paper, a fully automatic approach for the bifurcation classification in human Intravascular Optical Coherence Tomography (IV-OCT) sequences is introduced. Given the lumen contours, the method is capable of labeling the bifurcation slices. A geometric feature extraction was performed and the Forward Regression Orthogonal Least Squares method (FROLS) was applied to analyze the best features and to determine the appropriated weights in a binary classifier. A cross-validation scheme is applied in order to evaluate the performance of the classification approach and the results have shown a sensitivity of 86% and specificity of 92% to FROLS.

  4. A highly sensitive method for analyses of sugar moieties of glycoproteins by fluorescence labeling.

    PubMed

    Hase, S; Ikenaka, T; Matsushima, Y

    1981-08-01

    The sensitivity of a fluorescence labeling method ((1979). J. Biochem. 85, 989--994; 995--1002) for structure analyses of asparagine-linked sugar moieties of glycoproteins was increased by using HPLC with a fluorescence detector. Sugar moieties were separated from polypeptide portions by hydrazinolysis. Free amino groups thus exposed were acetylated and the reducing ends of sugar chains were reductively aminated with a fluorescent reagent, 2-aminopyridine, by the use of sodium cyanoborohydride. The pyridylamino derivatives were purified on a Dowex 1 column to eliminate undesired substances. The separation and identification of the pyridylamino derivatives were carried out by HPLC with a column of C18 reversed phase or gel permeation phase. As little as 0.1 pmol of pyridylamino derivatives can be detected. Ten microgram of Taka-amylase A was easily detected by this system. The method was also applied to some other glycoproteins.

  5. Adjustment method for microarray data generated using two-cycle RNA labeling protocol.

    PubMed

    Wang, Fugui; Chen, Rui; Ji, Dong; Bai, Shunong; Qian, Minping; Deng, Minghua

    2013-01-16

    Microarray technology is widely utilized for monitoring the expression changes of thousands of genes simultaneously. However, the requirement of relatively large amount of RNA for labeling and hybridization makes it difficult to perform microarray experiments with limited biological materials, thus leads to the development of many methods for preparing and amplifying mRNA. It is addressed that amplification methods usually bring bias, which may strongly hamper the following interpretation of the results. A big challenge is how to correct for the bias before further analysis. In this article, we observed the bias in rice gene expression microarray data generated with the Affymetrix one-cycle, two-cycle RNA labeling protocols, followed by validation with Real Time PCR. Based on these data, we proposed a statistical framework to model the processes of mRNA two-cycle linear amplification, and established a linear model for probe level correction. Maximum Likelihood Estimation (MLE) was applied to perform robust estimation of the Retaining Rate for each probe. After bias correction, some known pre-processing methods, such as PDNN, could be combined to finish preprocessing. Then, we evaluated our model and the results suggest that our model can effectively increase the quality of the microarray raw data: (i) Decrease the Coefficient of Variation for PM intensities of probe sets; (ii) Distinguish the microarray samples of five stages for rice stamen development more clearly; (iii) Improve the correlation coefficients among stamen microarray samples. We also discussed the necessity of model adjustment by comparing with another simple adjustment method. We conclude that the adjustment model is necessary and could effectively increase the quality of estimation for gene expression from the microarray raw data.

  6. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... covered by this section shall be process yellow or equivalent and process black. The label shall be printed full bleed process yellow. All type and graphics shall be print process black. (d) Label types... volume (VT) as determined in accordance § 305.7. (5) Unless otherwise indicated in this...

  7. Spiked proteomic standard dataset for testing label-free quantitative software and statistical methods.

    PubMed

    Ramus, Claire; Hovasse, Agnès; Marcellin, Marlène; Hesse, Anne-Marie; Mouton-Barbosa, Emmanuelle; Bouyssié, David; Vaca, Sebastian; Carapito, Christine; Chaoui, Karima; Bruley, Christophe; Garin, Jérôme; Cianférani, Sarah; Ferro, Myriam; Dorssaeler, Alain Van; Burlet-Schiltz, Odile; Schaeffer, Christine; Couté, Yohann; Gonzalez de Peredo, Anne

    2016-03-01

    This data article describes a controlled, spiked proteomic dataset for which the "ground truth" of variant proteins is known. It is based on the LC-MS analysis of samples composed of a fixed background of yeast lysate and different spiked amounts of the UPS1 mixture of 48 recombinant proteins. It can be used to objectively evaluate bioinformatic pipelines for label-free quantitative analysis, and their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. More specifically, it can be useful for tuning software tools parameters, but also testing new algorithms for label-free quantitative analysis, or for evaluation of downstream statistical methods. The raw MS files can be downloaded from ProteomeXchange with identifier PXD001819. Starting from some raw files of this dataset, we also provide here some processed data obtained through various bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold) in different workflows, to exemplify the use of such data in the context of software benchmarking, as discussed in details in the accompanying manuscript [1]. The experimental design used here for data processing takes advantage of the different spike levels introduced in the samples composing the dataset, and processed data are merged in a single file to facilitate the evaluation and illustration of software tools results for the detection of variant proteins with different absolute expression levels and fold change values.

  8. Spiked proteomic standard dataset for testing label-free quantitative software and statistical methods

    PubMed Central

    Ramus, Claire; Hovasse, Agnès; Marcellin, Marlène; Hesse, Anne-Marie; Mouton-Barbosa, Emmanuelle; Bouyssié, David; Vaca, Sebastian; Carapito, Christine; Chaoui, Karima; Bruley, Christophe; Garin, Jérôme; Cianférani, Sarah; Ferro, Myriam; Dorssaeler, Alain Van; Burlet-Schiltz, Odile; Schaeffer, Christine; Couté, Yohann; Gonzalez de Peredo, Anne

    2015-01-01

    This data article describes a controlled, spiked proteomic dataset for which the “ground truth” of variant proteins is known. It is based on the LC-MS analysis of samples composed of a fixed background of yeast lysate and different spiked amounts of the UPS1 mixture of 48 recombinant proteins. It can be used to objectively evaluate bioinformatic pipelines for label-free quantitative analysis, and their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. More specifically, it can be useful for tuning software tools parameters, but also testing new algorithms for label-free quantitative analysis, or for evaluation of downstream statistical methods. The raw MS files can be downloaded from ProteomeXchange with identifier PXD001819. Starting from some raw files of this dataset, we also provide here some processed data obtained through various bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold) in different workflows, to exemplify the use of such data in the context of software benchmarking, as discussed in details in the accompanying manuscript [1]. The experimental design used here for data processing takes advantage of the different spike levels introduced in the samples composing the dataset, and processed data are merged in a single file to facilitate the evaluation and illustration of software tools results for the detection of variant proteins with different absolute expression levels and fold change values. PMID:26862574

  9. Validation of the use of doubly labeled water for estimating metabolic rate in the green turtle (Chelonia mydas L.): a word of caution.

    PubMed

    Jones, T Todd; Hastings, Mervin D; Bostrom, Brian L; Andrews, Russel D; Jones, David R

    2009-08-01

    Marine turtles often have extremely high water turnover accompanied by a low field metabolic rate (FMR), a combination that can contraindicate the use of doubly labelled water (DLW). Therefore, we conducted a validation study to assess the suitability of the DLW technique for determining FMR of marine turtles. Six green turtles (22.42+/-3.13 kg) were injected with DLW and placed in a tank of seawater with a respirometer for continuous monitoring of oxygen consumption (MR) over a 5-day period. Trials were conducted for turtles in both fed and fasted states. Respiratory exchange ratio (RER) was determined in a dry respirometer and used to calculate energy expenditure. For fed and fasted turtles, total body water (TBW) was 66.67+/-3.37% and 58.70+/-7.63% of body mass, and water flux rates were 9.57+/-1.33% and 6.14+/-0.65% TBW day(-1), respectively. Water turnover in fasted turtles was 36% lower than that of fed turtles but MR (from oxygen consumption) of fasted turtles (13.77+/-1.49 kJ kg(-1) day(-1)) was 52% lower than in fed turtles (28.66+/-5.31 kJ kg(-1) day(-1)). Deuterium to oxygen-18 turnover rate (k(d):k(o)) ratios averaged 0.91+/-0.02 for fed turtles and 1.07+/-0.16 for fasted turtles. Fed turtles had a mean group difference of 8% and a mean individual difference of 53% between DLW and respirometry. The DLW method gave negative MR values in fasted turtles and could not be compared with respirometry data. Researchers should use caution when applying the DLW method in marine reptiles, especially when high water flux causes >90% of the labeled oxygen turnover to be due to water exchange.

  10. Radiometric method for determining solubility of organic solvents in water

    SciTech Connect

    Lo, J.M.; Tseng, C.L.; Yang, J.Y.

    1986-06-01

    Cobalt-60 labeled cobalt(III) pyrrolidinecarbodithioate (/sup 60/Co(PDC)/sub 3/) has a peculiar stability during storage in organic solvent and when its organic solution is shaken with an aqueous solution containing different acids or ions. Using these characteristics, the authors have attempted to use /sup 60/Co(PDC)/sub 3/ as a radioagent for determining solubilities of various organic solvents in water. The radioagent was first dissolved in the organic solvent under investigation before pure water was added. The solution mixture was shaken vigorously in order to let the organic phase contact with water sufficiently. Some of the organic solvent would dissolve in water after shaking, resulting in volume reduction of the organic phase. However, the radioagent was found not to accompany the organic solvent molecules going into water; i.e., all the radioactivity of /sup 60/Co(PDC)/sub 3/ would be retained in the organic phase. Solubility of the organic solvent in water therefore can be calculated from the value of the volume change of the organic phase divided by the water volume. Direct measurement of a small change in volume of organic phase with high accuracy is generally very difficult; alternatively, the authors have measured the specific activities of /sup 60/Co(PDC)/sub 3/ (cpm/mL) in the original and the final organic solutions, and the counting results were used to estimate the decrease in volume of the organic phase. Several commonly used organic solvents were selected to test the applicability of the proposed radiometric method. The solubilities of the organic solvents selected for this study range from very small values (10/sup -4/) to relatively large values (10/sup -2/), 6 references, 1 table.

  11. An Improved Method for Determining Absolute Phosphorylation Stoichiometry Using Bayesian Statistics and Isobaric Labeling.

    PubMed

    Lim, Matthew Y; O'Brien, Jonathon; Paulo, Joao A; Gygi, Steven P

    2017-10-06

    Phosphorylation stoichiometry, or occupancy, is one element of phosphoproteomics that can add useful biological context(1). We previously developed a method to assess phosphorylation stoichiometry on a proteome-wide scale(2). The stoichiometry calculation relies on identifying and measuring the levels of each nonphosphorylated counterpart peptide with and without phosphatase treatment. The method, however, is problematic in that low stoichiometry phosphopeptides can return negative stoichiometry values if measurement error is larger than the percent stoichiometry. Here, we have improved the stoichiometry method through the use of isobaric labeling with 10-plex TMT reagents. In this way, 5 phosphatase treated and 5 untreated samples are compared simultaneously so that each stoichiometry is represented by 5 ratio measurements with no missing values. We applied the method to determine basal stoichiometries of HCT116 cells growing in culture. With this method, we analyzed 5 biological replicates simultaneously with no need for phosphopeptide enrichment. Additionally, we developed a Bayesian model to estimate phosphorylation stoichiometry as a parameter confined to an interval between 0 and 1 implemented as an R/Stan script. Consequently, both point and interval estimates are consistent with the plausible range of values for stoichiometry. Finally, we report absolute stoichiometry measurements with credible intervals for 6,772 phosphopeptides containing at least a single phosphorylation site.

  12. Label-free DNA hybridization detection by various spectroscopy methods using triphenylmethane dyes as a probe

    NASA Astrophysics Data System (ADS)

    Tu, Jiaojiao; Cai, Changqun; Ma, Ying; Luo, Lin; Weng, Chao; Chen, Xiaoming

    2012-12-01

    A new assay is developed for direct detection of DNA hybridization using triphenylmethane dye as a probe. It is based on various spectroscopic methods including resonance light scattering (RLS), circular dichroism (CD), ultraviolet spectra and fluorescence spectra, as well as atomic force microscopy (AFM), six triphenylmethane dyes interact with double strand DNA (dsDNA) and single strand DNA (ssDNA) were investigated, respectively. The interaction results in amplified resonance light scattering signals and enables the detection of hybridization without the need for labeling DNA. Mechanism investigations have shown that groove binding occurs between dsDNA and these triphenylmethane dyes, which depends on G-C sequences of dsDNA and the molecular volumes of triphenylmethane dyes. Our present approaches display the advantages of simple and fast, accurate and reliable, and the artificial samples were determined with satisfactory results.

  13. Label-free DNA hybridization detection by various spectroscopy methods using triphenylmethane dyes as a probe.

    PubMed

    Tu, Jiaojiao; Cai, Changqun; Ma, Ying; Luo, Lin; Weng, Chao; Chen, Xiaoming

    2012-12-01

    A new assay is developed for direct detection of DNA hybridization using triphenylmethane dye as a probe. It is based on various spectroscopic methods including resonance light scattering (RLS), circular dichroism (CD), ultraviolet spectra and fluorescence spectra, as well as atomic force microscopy (AFM), six triphenylmethane dyes interact with double strand DNA (dsDNA) and single strand DNA (ssDNA) were investigated, respectively. The interaction results in amplified resonance light scattering signals and enables the detection of hybridization without the need for labeling DNA. Mechanism investigations have shown that groove binding occurs between dsDNA and these triphenylmethane dyes, which depends on G-C sequences of dsDNA and the molecular volumes of triphenylmethane dyes. Our present approaches display the advantages of simple and fast, accurate and reliable, and the artificial samples were determined with satisfactory results. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Predicting free-living energy expenditure using a miniaturized ear-worn sensor: an evaluation against doubly labeled water.

    PubMed

    Bouarfa, Loubna; Atallah, Louis; Kwasnicki, Richard Mark; Pettitt, Claire; Frost, Gary; Yang, Guang-Zhong

    2014-02-01

    Accurate estimation of daily total energy expenditure (EE)is a prerequisite for assisted weight management and assessing certain health conditions. The use of wearable sensors for predicting free-living EE is challenged by consistent sensor placement, user compliance, and estimation methods used. This paper examines whether a single ear-worn accelerometer can be used for EE estimation under free-living conditions.An EE prediction model as first derived and validated in a controlled setting using healthy subjects involving different physical activities. Ten different activities were assessed showing a tenfold cross validation error of 0.24. Furthermore, the EE prediction model shows a mean absolute deviation(MAD) below 1.2 metabolic equivalent of tasks. The same model was applied to a free-living setting with a different population for further validation. The results were compared against those derived from doubly labeled water. In free-living settings, the predicted daily EE has a correlation of 0.74, p 0.008, and a MAD of 272 kcal day. These results demonstrate that laboratory-derived prediction models can be used to predict EE under free-living conditions [corrected].

  15. A new model for predicting energy requirements of children during catch-up growth developed using doubly labeled water.

    PubMed

    Fjeld, C R; Schoeller, D A; Brown, K H

    1989-05-01

    Energy partitioned to maintenance plus activity, tissue synthesis, and storage was measured in 41 children in early recovery [W/L (wt/length) less than 5th percentile] from severe protein-energy malnutrition and in late recovery (W/L = 25th percentile) to determine energy requirements during catch-up growth. Metabolizable energy intake was measured by bomb calorimetry and metabolic collections. Energy expended (means +/- SD) for maintenance and activity estimated by the doubly labeled water method was 97 +/- 12 kcal/kg FFM (fat-free mass) in early recovery and 98 +/- 12 kcal/kg FFM in late recovery (p greater than 0.5). Energy stored was 5-6 kcal/g of wt gain. Tissue synthesis increased energy expenditure by 1 +/- 0.7 kcal/g gain in both early and late recovery. From these data a mathematical model was developed to predict energy requirements for children during catch-up growth as a function of initial body composition and rate and composition of wt gain. The model for predicting metabolizable energy requirements is [(98 x FFM + A (11.1 B + 2.2 C)], kcal/kg.d, where FFM is fat-free mass expressed as a percentage of body wt, A is wt gain (g/kg.d), B and C are percentage of wt gain/100 as fat and FFM, respectively. The model was tested retrospectively in separate studies of malnourished children.

  16. Assessing Energy Requirements in Women With Polycystic Ovary Syndrome: A Comparison Against Doubly Labeled Water.

    PubMed

    Broskey, Nicholas T; Klempel, Monica C; Gilmore, L Anne; Sutton, Elizabeth F; Altazan, Abby D; Burton, Jeffrey H; Ravussin, Eric; Redman, Leanne M

    2017-06-01

    Weight loss is prescribed to offset the deleterious consequences of polycystic ovary syndrome (PCOS), but a successful intervention requires an accurate assessment of energy requirements. Describe energy requirements in women with PCOS and evaluate common prediction equations compared with doubly labeled water (DLW). Cross-sectional study. Academic research center. Twenty-eight weight-stable women with PCOS completed a 14-day DLW study along with measures of body composition and resting metabolic rate and assessment of physical activity by accelerometry. Total daily energy expenditure (TDEE) determined by DLW. TDEE was 2661 ± 373 kcal/d. TDEE estimated from four commonly used equations was within 4% to 6% of the TDEE measured by DLW. Hyperinsulinemia (fasting insulin and homeostatic model assessment of insulin resistance) was associated with TDEE estimates from all prediction equations (both r = 0.45; P = 0.02) but was not a significant covariate in a model that predicts TDEE. Similarly, hyperandrogenemia (total testosterone, free androgen index, and dehydroepiandrosterone sulfate) was not associated with TDEE. In weight-stable women with PCOS, the following equation derived from DLW can be used to determine energy requirements: TDEE (kcal/d) = 438 - [1.6 * Fat Mass (kg)] + [35.1 * Fat-Free Mass (kg)] + [16.2 * Age (y)]; R2 = 0.41; P = 0.005. Established equations using weight, height, and age performed well for predicting energy requirements in weight-stable women with PCOS, but more precise estimates require an accurate assessment of physical activity. Our equation derived from DLW data, which incorporates habitual physical activity, can also be used in women with PCOS; however, additional studies are needed for model validation.

  17. Evaluating real-time immunohistochemistry on multiple tissue samples, multiple targets and multiple antibody labeling methods

    PubMed Central

    2013-01-01

    Background Immunohistochemistry (IHC) is a well-established method for the analysis of protein expression in tissue specimens and constitutes one of the most common methods performed in pathology laboratories worldwide. However, IHC is a multi-layered method based on subjective estimations and differences in staining and interpretation has been observed between facilities, suggesting that the analysis of proteins on tissue would benefit from protocol optimization and standardization. Here we describe how the emerging and operator independent tool of real-time immunohistochemistry (RT-IHC) reveals a time resolved description of antibody interacting with target protein in formalin fixed paraffin embedded tissue. The aim was to understand the technical aspects of RT-IHC, regarding generalization of the concept and to what extent it can be considered a quantitative method. Results Three different antibodies labeled with fluorescent or radioactive labels were applied on nine different tissue samples from either human or mouse, and the results for all RT-IHC analyses distinctly show that the method is generally applicable. The collected binding curves showed that the majority of the antibody-antigen interactions did not reach equilibrium within 3 hours, suggesting that standardized protocols for immunohistochemistry are sometimes inadequately optimized. The impact of tissue size and thickness as well as the position of the section on the glass petri dish was assessed in order for practical details to be further elucidated for this emerging technique. Size and location was found to affect signal magnitude to a larger extent than thickness, but the signal from all measurements were still sufficient to trace the curvature. The curvature, representing the kinetics of the interaction, was independent of thickness, size and position and may be a promising parameter for the evaluation of e.g. biopsy sections of different sizes. Conclusions It was found that RT-IHC can be used

  18. Evaluating real-time immunohistochemistry on multiple tissue samples, multiple targets and multiple antibody labeling methods.

    PubMed

    Dubois, Louise; Andersson, Karl; Asplund, Anna; Björkelund, Hanna

    2013-12-18

    Immunohistochemistry (IHC) is a well-established method for the analysis of protein expression in tissue specimens and constitutes one of the most common methods performed in pathology laboratories worldwide. However, IHC is a multi-layered method based on subjective estimations and differences in staining and interpretation has been observed between facilities, suggesting that the analysis of proteins on tissue would benefit from protocol optimization and standardization. Here we describe how the emerging and operator independent tool of real-time immunohistochemistry (RT-IHC) reveals a time resolved description of antibody interacting with target protein in formalin fixed paraffin embedded tissue. The aim was to understand the technical aspects of RT-IHC, regarding generalization of the concept and to what extent it can be considered a quantitative method. Three different antibodies labeled with fluorescent or radioactive labels were applied on nine different tissue samples from either human or mouse, and the results for all RT-IHC analyses distinctly show that the method is generally applicable. The collected binding curves showed that the majority of the antibody-antigen interactions did not reach equilibrium within 3 hours, suggesting that standardized protocols for immunohistochemistry are sometimes inadequately optimized. The impact of tissue size and thickness as well as the position of the section on the glass petri dish was assessed in order for practical details to be further elucidated for this emerging technique. Size and location was found to affect signal magnitude to a larger extent than thickness, but the signal from all measurements were still sufficient to trace the curvature. The curvature, representing the kinetics of the interaction, was independent of thickness, size and position and may be a promising parameter for the evaluation of e.g. biopsy sections of different sizes. It was found that RT-IHC can be used for the evaluation of a number

  19. A label fusion method using conditional random fields with higher-order potentials: Application to hippocampal segmentation.

    PubMed

    Platero, Carlos; Carmen Tobar, M

    2015-06-01

    The objective of this study is to develop a probabilistic modeling framework for segmenting structures of interest from a collection of atlases. We present a label fusion method that is based on minimizing an energy function using graph-cut techniques. We use a conditional random field (CRF) model that allows us to efficiently incorporate shape, appearance and context information. This model is characterized by a pseudo-Boolean function defined on unary, pairwise and higher-order potentials. Given a subset of registered atlases in the target image for a particular region of interest (ROI), we first derive an appearance-shape model from these registered atlases. The unary potentials combine an appearance model based on multiple features with a label prior using a weighted voting method. The pairwise terms are defined from a Finsler metric that minimizes the surface of separation between voxels whose labels are different. The higher-order potentials used in our framework are based on the robust P(n) model proposed by Kohli et al. The higher-order potentials enforce label consistency in cliques; hence, the proposed method can be viewed as an approach to integrate high-level information with images based on low-level features. To evaluate the performance and the robustness of the proposed label fusion method, we employ two available databases of T1-weighted (T1W) magnetic resonance (MR) images of human brains. We compare our approach with other label fusion methods in the automatic hippocampal segmentation from T1W-MR images. Our label fusion method yields mean Dice coefficients of 0.829 and 0.790 for the two databases used with mean times of approximately 80 and 160s, respectively. We introduce a new label fusion method based on a CRF model and on ROIs. The CRF model is characterized by a pseudo-Boolean function defined on unary, pairwise and higher-order potentials. The proposed Boolean function is representable by graphs. A globally optimal binary labeling is found

  20. Rapid and simple methods for labeling white blood cells and platelets with indium-111-oxine

    SciTech Connect

    Steffel, F.G.; Rao, S.A.

    1987-06-01

    Simple procedures in a kit form for labeling white blood cells (WBCs) and platelets with commercially available indium-111 (/sup 111/In)-oxine have been developed for the convenience of small community hospitals. The time required for the labeling procedure is less than 2 hr. The resulting scintigrams from the clinical studies in both WBCs and platelets showed that the /sup 111/In-labeled cells have a high degree of viability.

  1. Water treatment: Chitosan associated with electrochemical methods

    NASA Astrophysics Data System (ADS)

    Tamiasso-Martinhon, Priscila; Marques Teixeira de Souza, João; Cruzeiro da Silva, Silvia Maria; Pellegrini Pessoa, Fernando Luiz; Sousa, Célia

    2017-04-01

    Pollution of water bodies due to the presence of toxic metals and organic compounds, bring out a series of environmental problems of public, government and social character. In addition, water pollution, has become the target and source of concern in many industrial sectors. Therefore, it is essential to develop technologies for treatment and purification of water. Chitosan is a natural product derived from chitin, extracted mainly from the shells of crustaceans. It is a low cost, renewable and biodegradable biopolymer of great socioeconomic and environmental importance. The classic treatment of wastewater containing metals involves physical chemistry processes of precipitation, ion exchange and electrochemistry. Electrochemical technology has been presented as the most promising methods for treating wastewater polluted with metals, colloids, dyes or oil in water emulsions; besides being used in removing organic compounds. Alternative methods like adsorption with biosorbents have been investigated. The great advantage of this latter over other techniques is the low generation of residues, easy recovery of metals and the possibility of reuse of the adsorbent. This article aimed to carry out an exploratory study, of bibliographical nature, on the use of chitosan in electrochemical methods for water treatment.

  2. A simplified method for peptide de novo sequencing using (18)O labeling.

    PubMed

    Voráĉ, Aleš; Sedo, Ondrej; Havliš, Jan; Zdráhal, Zbyněk

    2014-01-01

    Incorporation of an (18)O atom into a peptide C-terminus by proteolytic cleavage in the presence of H2(18)O is one of the most effective ways of enhancing tandem mass spectrometry (MS/MS)-based de novo sequencing. Incorporation is usually accomplished by procedures including vacuum-assisted drying of tryptic peptides extracted from gels, their subsequent reconstitution in a H2(16)O/H2(18)O mixture and re-treatment with trypsin. In the present work, we propose a simplified procedure for (18)O incorporation into tryptic peptides by adding H2(18)O and trypsin to the original digest solution. In comparison to published methods, the proposed protocol for peptide de novo sequencing brings significant advantages in analysis and workflow with no deterioration in method performance. We show that labeling by this simplified method leads to a highlighting of the y-ion fragment series in the peptide matrix-assisted laser desorption/ionization (MALDI)- MS/MS data, which facilitates MS/MS data interpretation. We also prove that eliminating acid extraction of peptides from gels does not result in a decrease in sequence coverage or a qualitative loss of particular peptides detectable by MALDI-MS. The method was examined by MALDI-MS/MS on bovine serum albumin and recombinant histidine kinase CKI1 from Arabidopsis thaliana, and was verified by de novo sequencing of tryptic peptides originating from Apodemus sylvaticus salivary proteins.

  3. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... size dimensions for all labels shall be as follows: width must be between 5 1/4 inches and 5 1/2 inches (13.34 cm. and 13.97 cm.); length must be between 7 3/8 inches (18.78 cm.) and 7 5/8 (19.34 cm.). Copy... process yellow or equivalent and process black. The label shall be printed full bleed process yellow....

  4. Selective spectroscopic methods for water analysis

    SciTech Connect

    Vaidya, Bikas

    1997-06-24

    This dissertation explores in large part the development of a few types of spectroscopic methods in the analysis of water. Methods for the determination of some of the most important properties of water like pH, metal ion content, and chemical oxygen demand are investigated in detail. This report contains a general introduction to the subject and the conclusions. Four chapters and an appendix have been processed separately. They are: chromogenic and fluorogenic crown ether compounds for the selective extraction and determination of Hg(II); selective determination of cadmium in water using a chromogenic crown ether in a mixed micellar solution; reduction of chloride interference in chemical oxygen demand determination without using mercury salts; structural orientation patterns for a series of anthraquinone sulfonates adsorbed at an aminophenol thiolate monolayer chemisorbed at gold; and the role of chemically modified surfaces in the construction of miniaturized analytical instrumentation.

  5. Comparing model-based and model-free analysis methods for QUASAR arterial spin labeling perfusion quantification.

    PubMed

    Chappell, Michael A; Woolrich, Mark W; Petersen, Esben T; Golay, Xavier; Payne, Stephen J

    2013-05-01

    Amongst the various implementations of arterial spin labeling MRI methods for quantifying cerebral perfusion, the QUASAR method is unique. By using a combination of labeling with and without flow suppression gradients, the QUASAR method offers the separation of macrovascular and tissue signals. This permits local arterial input functions to be defined and "model-free" analysis, using numerical deconvolution, to be used. However, it remains unclear whether arterial spin labeling data are best treated using model-free or model-based analysis. This work provides a critical comparison of these two approaches for QUASAR arterial spin labeling in the healthy brain. An existing two-component (arterial and tissue) model was extended to the mixed flow suppression scheme of QUASAR to provide an optimal model-based analysis. The model-based analysis was extended to incorporate dispersion of the labeled bolus, generally regarded as the major source of discrepancy between the two analysis approaches. Model-free and model-based analyses were compared for perfusion quantification including absolute measurements, uncertainty estimation, and spatial variation in cerebral blood flow estimates. Major sources of discrepancies between model-free and model-based analysis were attributed to the effects of dispersion and the degree to which the two methods can separate macrovascular and tissue signal. Copyright © 2012 Wiley Periodicals, Inc.

  6. Methods for Chemical Analysis of Fresh Waters.

    ERIC Educational Resources Information Center

    Golterman, H. L.

    This manual, one of a series prepared for the guidance of research workers conducting studies as part of the International Biological Programme, contains recommended methods for the analysis of fresh water. The techniques are grouped in the following major sections: Sample Taking and Storage; Conductivity, pH, Oxidation-Reduction Potential,…

  7. Methods for Chemical Analysis of Fresh Waters.

    ERIC Educational Resources Information Center

    Golterman, H. L.

    This manual, one of a series prepared for the guidance of research workers conducting studies as part of the International Biological Programme, contains recommended methods for the analysis of fresh water. The techniques are grouped in the following major sections: Sample Taking and Storage; Conductivity, pH, Oxidation-Reduction Potential,…

  8. A Comparison of Methods for Teaching Receptive Labeling to Children with Autism Spectrum Disorders: A Systematic Replication

    ERIC Educational Resources Information Center

    Grow, Laura L.; Kodak, Tiffany; Carr, James E.

    2014-01-01

    Previous research has demonstrated that the conditional-only method (starting with a multiple-stimulus array) is more efficient than the simple-conditional method (progressive incorporation of more stimuli into the array) for teaching receptive labeling to children with autism spectrum disorders (Grow, Carr, Kodak, Jostad, & Kisamore, 2011).…

  9. Points of Convergence in Music Education: The Use of Data Labels as a Strategy for Mixed Methods Integration

    ERIC Educational Resources Information Center

    Fitzpatrick, Kate R.

    2016-01-01

    Although the mixing of quantitative and qualitative data is an essential component of mixed methods research, the process of integrating both types of data in meaningful ways can be challenging. The purpose of this article is to describe the use of data labels in mixed methods research as a technique for the integration of qualitative and…

  10. Points of Convergence in Music Education: The Use of Data Labels as a Strategy for Mixed Methods Integration

    ERIC Educational Resources Information Center

    Fitzpatrick, Kate R.

    2016-01-01

    Although the mixing of quantitative and qualitative data is an essential component of mixed methods research, the process of integrating both types of data in meaningful ways can be challenging. The purpose of this article is to describe the use of data labels in mixed methods research as a technique for the integration of qualitative and…

  11. Method for sequential staining of GTL-banded metaphases with fluorescent-labeled chromosome-specific paint probes.

    PubMed

    Jalal, S M; Law, M E; Christensen, E R; Spurbeck, J L; Dewald, G W

    1993-04-01

    We describe a method for use of fluorescent-labeled whole chromosome-specific paint probes on GTL-banded metaphases to utilize the combined potential of these techniques for defining chromosome abnormalities. The efficacy of this method was tested on 6 cases involving different chromosome abnormalities and various tissues, including blood, amniotic fluid, skin fibroblasts, and bone marrow.

  12. Surface Functionalization Methods to Enhance Bioconjugation in Metal-Labeled Polystyrene Particles

    PubMed Central

    Abdelrahman, Ahmed I.; Thickett, Stuart C.; Liang, Yi; Ornatsky, Olga; Baranov, Vladimir; Winnik, Mitchell A.

    2011-01-01

    Lanthanide-encoded polystyrene particles synthesized by dispersion polymerization are excellent candidates for mass cytometry based immunoassays, however they have previously lacked the ability to conjugate biomolecules to the particle surface. We present here three approaches to post-functionalize these particles, enabling the covalent attachment of proteins. Our first approach used partially hydrolyzed poly(N-vinylpyrrolidone) as a dispersion polymerization stabilizer to synthesize particles with high concentration of -COOH groups on the particle surface. In an alternative strategy to provide -COOH functionality to the lanthanide-encoded particles, we employed seeded emulsion polymerization to graft poly(methacrylic acid) (PMAA) chains onto the surface of these particles. However, these two approaches gave little to no improvement in the extent of bioconjugation. In our third approach, seeded emulsion polymerization was subsequently used as a method to grow a functional polymer shell (in this case, poly(glycidyl methacrylate) (PGMA)) onto the surface of these particles, which proved highly successful. The epoxide-rich PGMA shell permitted extensive surface bioconjugation of NeutrAvidin, as probed by an Lu-labeled biotin reporter (ca. 7 × 105 binding events per particle with a very low amount of non-specific binding) and analyzed by mass cytometry. It was shown that coupling agents such as EDC were not needed, such was the reactivity of the particle surface. These particles were stable and the addition of a polymeric shell was shown did not affect the narrow lanthanide ion distribution within the particle interior as analyzed by mass cytometry. These particles represent the most promising candidates for the development of a highly multiplexed bioassay based on lanthanide-labeled particles to date. PMID:21799543

  13. A novel label-free fluorescence assay for one-step sensitive detection of Hg2+ in environmental drinking water samples

    PubMed Central

    Li, Ya; Liu, Nan; Liu, Hui; Wang, Yu; Hao, Yuwei; Ma, Xinhua; Li, Xiaoli; Huo, Yapeng; Lu, Jiahai; Tang, Shuge; Wang, Caiqin; Zhang, Yinhong; Gao, Zhixian

    2017-01-01

    A novel label-free fluorescence assay for detection of Hg2+ was developed based on the Hg2+-binding single-stranded DNA (ssDNA) and SYBR Green I (SG I). Differences from other assays, the designed rich-thymine (T) ssDNA probe without fluorescent labelling can be rapidly formed a T-Hg2+-T complex and folded into a stable hairpin structure in the presence of Hg2+ in environmental drinking water samples by facilitating fluorescence increase through intercalating with SG I in one-step. In the assay, the fluorescence signal can be directly obtained without additional incubation within 1 min. The dynamic quantitative working ranges was 5–1000 nM, the determination coefficients were satisfied by optimization of the reaction conditions. The lowest detection limit of Hg2+ was 3 nM which is well below the standard of U.S. Environmental Protection Agency. This method was highly specific for detecting of Hg2+ without being affected by other possible interfering ions from different background compositions of water samples. The recoveries of Hg2+ spiked in these samples were 95.05–103.51%. The proposed method is more viable, low-costing and simple for operation in field detection than the other methods with great potentials, such as emergency disposal, environmental monitoring, surveillance and supporting of ecological risk assessment and management. PMID:28378768

  14. A novel label-free fluorescence assay for one-step sensitive detection of Hg2+ in environmental drinking water samples

    NASA Astrophysics Data System (ADS)

    Li, Ya; Liu, Nan; Liu, Hui; Wang, Yu; Hao, Yuwei; Ma, Xinhua; Li, Xiaoli; Huo, Yapeng; Lu, Jiahai; Tang, Shuge; Wang, Caiqin; Zhang, Yinhong; Gao, Zhixian

    2017-04-01

    A novel label-free fluorescence assay for detection of Hg2+ was developed based on the Hg2+-binding single-stranded DNA (ssDNA) and SYBR Green I (SG I). Differences from other assays, the designed rich-thymine (T) ssDNA probe without fluorescent labelling can be rapidly formed a T-Hg2+-T complex and folded into a stable hairpin structure in the presence of Hg2+ in environmental drinking water samples by facilitating fluorescence increase through intercalating with SG I in one-step. In the assay, the fluorescence signal can be directly obtained without additional incubation within 1 min. The dynamic quantitative working ranges was 5-1000 nM, the determination coefficients were satisfied by optimization of the reaction conditions. The lowest detection limit of Hg2+ was 3 nM which is well below the standard of U.S. Environmental Protection Agency. This method was highly specific for detecting of Hg2+ without being affected by other possible interfering ions from different background compositions of water samples. The recoveries of Hg2+ spiked in these samples were 95.05-103.51%. The proposed method is more viable, low-costing and simple for operation in field detection than the other methods with great potentials, such as emergency disposal, environmental monitoring, surveillance and supporting of ecological risk assessment and management.

  15. A novel label-free fluorescence assay for one-step sensitive detection of Hg(2+) in environmental drinking water samples.

    PubMed

    Li, Ya; Liu, Nan; Liu, Hui; Wang, Yu; Hao, Yuwei; Ma, Xinhua; Li, Xiaoli; Huo, Yapeng; Lu, Jiahai; Tang, Shuge; Wang, Caiqin; Zhang, Yinhong; Gao, Zhixian

    2017-04-05

    A novel label-free fluorescence assay for detection of Hg(2+) was developed based on the Hg(2+)-binding single-stranded DNA (ssDNA) and SYBR Green I (SG I). Differences from other assays, the designed rich-thymine (T) ssDNA probe without fluorescent labelling can be rapidly formed a T-Hg(2+)-T complex and folded into a stable hairpin structure in the presence of Hg(2+) in environmental drinking water samples by facilitating fluorescence increase through intercalating with SG I in one-step. In the assay, the fluorescence signal can be directly obtained without additional incubation within 1 min. The dynamic quantitative working ranges was 5-1000 nM, the determination coefficients were satisfied by optimization of the reaction conditions. The lowest detection limit of Hg(2+) was 3 nM which is well below the standard of U.S. Environmental Protection Agency. This method was highly specific for detecting of Hg(2+) without being affected by other possible interfering ions from different background compositions of water samples. The recoveries of Hg(2+) spiked in these samples were 95.05-103.51%. The proposed method is more viable, low-costing and simple for operation in field detection than the other methods with great potentials, such as emergency disposal, environmental monitoring, surveillance and supporting of ecological risk assessment and management.

  16. Water Polo Injuries and Training Methods.

    PubMed

    Spittler, Jack; Keeling, James

    Water polo is a unique team sport combining swimming sprints and eggbeater kicking, frequent overhead movements and throwing, and regular physical contact with minimal protective equipment. Accordingly, a wide variety of training methods attempt to enhance all of these skill sets. This usually includes some combination of aerobic/anaerobic fitness (via swimming), sport-specific skills, strengthening, and nutrition. In addition, injuries in water polo are somewhat diverse. Physical contact is responsible for the majority of acute injuries, most frequently being injuries to the head and face. The high prevalence of shoulder pain in water polo is likely related to increased shoulder mobility and subsequent instability and stress on shoulder structures, yet the underlying causation is not certain. The unique aspect of shoulder injuries occurring in water polo players is that they may be due to a combination of swimming-related overuse conditions, overhead throwing, and acute trauma-related conditions. Although there is generally minimal evidence-based information available, this article attempts to highlight the current knowledge that we have in regard to water polo injuries and training methods.

  17. Methods for the synthesis of tritium-labelled fatty acids and their derivatives, oxylipins and steroids

    NASA Astrophysics Data System (ADS)

    Shevchenko, Valerii P.; Nagaev, Igor Yu; Myasoedov, Nikolai F.

    1999-10-01

    The achievements in the field of synthesis and application of tritium-labelled oxylipins, steroids, fatty acids, phospho-, sphingo- and other lipids are reviewed. The importance of these studies for the solution of current problems of biochemistry, biology and pharmacology is exemplified in the application of labelled compounds. The bibliography includes 148 references.

  18. Mass Spectrometric Quantification of Histone Post-translational Modifications by a Hybrid Chemical Labeling Method

    PubMed Central

    Maile, Tobias M.; Izrael-Tomasevic, Anita; Cheung, Tommy; Guler, Gulfem D.; Tindell, Charles; Masselot, Alexandre; Liang, Jun; Zhao, Feng; Trojer, Patrick; Classon, Marie; Arnott, David

    2015-01-01

    Mass spectrometry is a powerful alternative to antibody-based methods for the analysis of histone post-translational modifications (marks). A key development in this approach was the deliberate propionylation of histones to improve sequence coverage across the lysine-rich and hydrophilic tails that bear most modifications. Several marks continue to be problematic however, particularly di- and tri-methylated lysine 4 of histone H3 which we found to be subject to substantial and selective losses during sample preparation and liquid chromatography-mass spectrometry. We developed a new method employing a “one-pot” hybrid chemical derivatization of histones, whereby an initial conversion of free lysines to their propionylated forms under mild aqueous conditions is followed by trypsin digestion and labeling of new peptide N termini with phenyl isocyanate. High resolution mass spectrometry was used to collect qualitative and quantitative data, and a novel web-based software application (Fishtones) was developed for viewing and quantifying histone marks in the resulting data sets. Recoveries of 53 methyl, acetyl, and phosphoryl marks on histone H3.1 were improved by an average of threefold overall, and over 50-fold for H3K4 di- and tri-methyl marks. The power of this workflow for epigenetic research and drug discovery was demonstrated by measuring quantitative changes in H3K4 trimethylation induced by small molecule inhibitors of lysine demethylases and siRNA knockdown of epigenetic modifiers ASH2L and WDR5. PMID:25680960

  19. A rapid method for manual or automated purification of fluorescently labeled nucleic acids for sequencing, genotyping, and microarrays.

    PubMed

    Springer, Amy L; Booth, Lisa R; Braid, Michael D; Houde, Christiane M; Hughes, Karin A; Kaiser, Robert J; Pedrak, Casandra; Spicer, Douglas A; Stolyar, Sergey

    2003-03-01

    Fluorescent dyes provide specific, sensitive, and multiplexed detection of nucleic acids. To maximize sensitivity, fluorescently labeled reaction products (e.g., cycle sequencing or primer extension products) must be purified away from residual dye-labeled precursors. Successful high-throughput analyses require that this purification be reliable, rapid, and amenable to automation. Common methods for purifying reaction products involve several steps and require processes that are not easily automated. Prolinx, Inc. has devel oped RapXtract superparamagnetic separation technology affording rapid and easy-to-perform methods that yield high-quality product and are easily automated. The technology uses superparamagnetic particles that specifically remove unincorporated dye-labeled precursors. These particles are efficiently pelleted in the presence of a magnetic field, making them ideal for purification because of the rapid separations that they allow. RapXtract-purified sequencing reactions yield data with good signal and high Phred quality scores, and they work with various sequencing dye chemistries, including BigDye and near-infrared fluorescence IRDyes. RapXtract technology can also be used to purify dye primer sequencing reactions, primer extension reactions for genotyping analysis, and nucleic acid labeling reactions for microarray hybridization. The ease of use and versatility of RapXtract technology makes it a good choice for manual or automated purification of fluorescently labeled nucleic acids.

  20. Assesment of perfusion in glial tumors with arterial spin labeling; comparison with dynamic susceptibility contrast method.

    PubMed

    Cebeci, H; Aydin, O; Ozturk-Isik, E; Gumus, C; Inecikli, F; Bekar, A; Kocaeli, H; Hakyemez, B

    2014-10-01

    Arterial spin labeling perfusion imaging (ASL-PI) is a non-invasive perfusion imaging method that can be used for evaluation and quantification of cerebral blood flow (CBF). Aim of our study was to evaluating the efficiency of ASL in histopathological grade estimation of glial tumors and comparing findings with dynamic susceptibility contrast perfusion imaging (DSC-PI) method. This study involved 33 patients (20 high-grade and 13 low-grade gliomas). Multiphase multislice pulsed ASL MRI sequence and a first-passage gadopentetate dimeglumine T2*-weighted gradient-echo single-shot echo-planar sequence were acquired for all the patients. For each patient, perfusion relative signal intensity (rSI), CBF and relative CBF (rCBF) on ASL-PI and relative cerebral blood volume (rCBV) and relative cerebral blood flow (rCBF) values on DSC-PI were determined. The relative signal intensity of each tumor was determined as the maximal SI within the tumor divided by SI within symetric region in the contralateral hemisphere on ASL-PI. rCBV and rCBF were calculated by deconvolution of an arterial input function. Relative values of the lesions were obtained by dividing the values to the normal appearing symmetric region on the contralateral hemisphere. For statistical analysis, Mann-Whitney ranksum test was carried out. Receiver operating characteristic curve (ROC) analysis was performed to assess the relationship between the rCBF-ASL, rSI-ASL, rCBV and rCBF ratios and grade of gliomas. Their cut-off values permitting best discrimination was calculated. The correlation between rCBV, rCBF, rSI-ASL and rCBF-ASL and glioma grade was assessed using Spearman correlation analysis. There was a statistically significant difference between low and high-grade tumors for all parameters. Correlation analyses revealed significant positive correlations between rCBV and rCBF-ASL (r=0.81, p<0.001). However correlation between rCBF and rCBF-ASL was weaker (r=0.64, p<0.001). Arterial spin labeling is an

  1. Method of removing oxidized contaminants from water

    DOEpatents

    Amonette, James E.; Fruchter, Jonathan S.; Gorby, Yuri A.; Cole, Charles R.; Cantrell, Kirk J.; Kaplan, Daniel I.

    1998-01-01

    The present invention is a method for removing oxidized contaminant(s) from water. More specifically, the invention has the steps of contacting water containing the oxidized contaminant(s) with a layered aluminosilicate having Fe(II). The aluminosilicate may contain naturally occurring Fe(II), or the Fe(II) may be produced by reducing Fe(III) that is initially present. Reduction may be either by exposure to a chemical or biological reductant. Contacting the water containing oxidized contaminant(s) may be by (1) injection of Fe(II)-containing layered aluminosilicate, via a well, into a saturated zone where it is likely to intercept the contaminated water; (2) injection of contaminated water into a vessel containing the Fe(II)-bearing layered aluminosilicate; and (3) first reducing Fe(III) in the layered aluminosilicate to Fe(II) by injection of a biological or chemical reductant, into an aquifer or vessel having sufficient Fe(III)-bearing aluminosilicate to produce the necessary Fe(II).

  2. Method of removing oxidized contaminants from water

    DOEpatents

    Amonette, J.E.; Fruchter, J.S.; Gorby, Y.A.; Cole, C.R.; Cantrell, K.J.; Kaplan, D.I.

    1998-07-21

    The present invention is a method for removing oxidized contaminant(s) from water. More specifically, the invention has the steps of contacting water containing the oxidized contaminant(s) with a layered aluminosilicate having Fe(II). The aluminosilicate may contain naturally occurring Fe(II), or the Fe(II) may be produced by reducing Fe(III) that is initially present. Reduction may be either by exposure to a chemical or biological reductant. Contacting the water containing oxidized contaminant(s) may be by (1) injection of Fe(II)-containing layered aluminosilicate, via a well, into a saturated zone where it is likely to intercept the contaminated water; (2) injection of contaminated water into a vessel containing the Fe(II)-bearing layered aluminosilicate; and (3) first reducing Fe(III) in the layered aluminosilicate to Fe(II) by injection of a biological or chemical reductant, into an aquifer or vessel having sufficient Fe(III)-bearing aluminosilicate to produce the necessary Fe(II). 8 figs.

  3. Extraction and labeling methods for microarrays using small amounts of plant tissue.

    PubMed

    Stimpson, Alexander J; Pereira, Rhea S; Kiss, John Z; Correll, Melanie J

    2009-03-01

    Procedures were developed to maximize the yield of high-quality RNA from small amounts of plant biomass for microarrays. Two disruption techniques (bead milling and pestle and mortar) were compared for the yield and the quality of RNA extracted from 1-week-old Arabidopsis thaliana seedlings (approximately 0.5-30 mg total biomass). The pestle and mortar method of extraction showed enhanced RNA quality at the smaller biomass samples compared with the bead milling technique, although the quality in the bead milling could be improved with additional cooling steps. The RNA extracted from the pestle and mortar technique was further tested to determine if the small quantity of RNA (500 ng-7 microg) was appropriate for microarray analyses. A new method of low-quantity RNA labeling for microarrays (NuGEN Technologies, Inc.) was used on five 7-day-old seedlings (approximately 2.5 mg fresh weight total) of Arabidopsis that were grown in the dark and exposed to 1 h of red light or continued dark. Microarray analyses were performed on a small plant sample (five seedlings; approximately 2.5 mg) using these methods and compared with extractions performed with larger biomass samples (approximately 500 roots). Many well-known light-regulated genes between the small plant samples and the larger biomass samples overlapped in expression changes, and the relative expression levels of selected genes were confirmed with quantitative real-time polymerase chain reaction, suggesting that these methods can be used for plant experiments where the biomass is extremely limited (i.e. spaceflight studies).

  4. Measurement of quantity of iron in magnetically labeled cells: comparison among different UV/VIS spectrometric methods.

    PubMed

    Rad, Ali M; Janic, Branislava; Iskander, A S M; Soltanian-Zadeh, Hamid; Arbab, Ali S

    2007-11-01

    Cell labeling with superparamagnetic iron oxides (SPIO) is becoming a routine procedure in cellular magnetic resonance imaging (MRI). Quantifying the intracellular iron in labeled cells is a prerequisite for determining the number of accumulated cells by quantitative MRI studies. To establish the most sensitive and reproducible method for measuring iron concentration in magnetically labeled cells, we investigated and compared four different methods using an ultraviolet-visible (UV/VIS) spectrophotometer. Background spectra were obtained for 5 and 10 M hydrochloric acids, a mixture of 100 mM citric acid plus ascorbic acid and bathophenanthroline sulphonate (BPS), and a mixture of 5 M hydrochloric acid plus 5% ferrocyanide. Spectra of the same solutions containing either 10 or 5 microg/mL iron oxides were also created to determine the peak absorbance wavelengths for the dissolved iron. In addition, different known iron concentrations were used to obtain calibration lines for each method. Based on the calibration factors, iron was measured in samples with a known amount of iron and in labeled cells. Methods based on the use of 10 M hydrochloric acid underestimated iron concentration in all experiments; for this method to give an accurate measurement, iron concentration in sample needs to be at least 3 microg/mL.

  5. Extracorporeal adsorption therapy: A Method to improve targeted radiation delivered by radiometal-labeled monoclonal antibodies.

    SciTech Connect

    Nemecek, Eneida R.; Green, Damian J.; Fisher, Darrell R.; Pagal, John M.; Lin, Yukang; Gopal, A. K.; Durack, Lawrence D.; Rajendran, Joseph G.; Wilbur, D. S.; Nilsson, Rune; Sandberg, Bengt; Press, Oliver W.

    2008-04-01

    Many investigators have demonstrated the ability to treat hematologic malignancies with radiolabeled monoclonal antibodies targeting hematopoietic antigens such as anti-CD20 and anti-CD45. [1-5] Although the remission rates achieved with radioimmunotherapy (RIT) are relatively high, many patients subsequently relapse presumably due to suboptimal delivery of enough radiation to eradicate the malignancy. The dose-response of leukemia and lymphoma to radiation has been proven. Substantial amounts of radiation can be delivered by RIT if followed by hematopoietic cell transplantation to rescue the bone marrow from myeloablation.[ref] However, the maximum dose of RIT that can be used is still limited by toxicity to normal tissues affected by nonspecific delivery of radiation. Efforts to improve RIT focus on improving the therapeutic ratios of radiation in target versus non-target tissues by removing the fraction of radioisotope that fails to bind to target tissues and circulates freely in the bloodstream perfusing non-target tissues. Our group and others have explored several alternatives for removal of unbound circulating antibody. [refs] One such method, extracorporeal adsorption therapy (ECAT) consists of removing unbound antibody by a method similar to plasmapheresis after critical circulation time and distribution of antibody into target tissues have been achieved. Preclinical studies of ECAT in murine xenograft models demonstrated significant improvement in therapeutic ratios of radioactivity. Chen and colleagues demonstrated that a 2-hour ECAT procedure could remove 40 to 70% of the radioactivity from liver, lung and spleen. [ref] Although isotope concentration in the tumor was initially unaffected, a 50% decrease was noted approximately 36 hours after the procedure. This approach was also evaluated in a limited phase I pilot study of patients with refractory B-cell lymphoma. [ref] After radiographic confirmation of tumor localization of a test dose of anti-CD20

  6. Study of the temperature influence on catalase using spin labelling method

    NASA Astrophysics Data System (ADS)

    Bartoszek, M.; Kściuczyk, M.

    2005-06-01

    Electron paramagnetic resonance (EPR) spectroscopy has been used to study the temperature influence on spin labelled catalase. The measurements were made in the temperature range 300-345 K. The spin label technique allows to observe the structural changes in catalase with increasing temperature. The rotational correlation time of the 3-(2-iodoacetamido)-proxyl spin marker placed in metalloenzyme was determined. The details of ESR spectra contain information on the character of the spin label motion. It indicates the changes in the structure of catalase before the denaturation temperature, determined with dsc microcalorimetry.

  7. Competitive labelling, a method for determining the reactivity of individual groups in proteins. The amino groups of porcine elastase

    PubMed Central

    Kaplan, H.; Stevenson, K. J.; Hartley, B. S.

    1971-01-01

    1. A method is described for determining the ionization constants and reactivities of individual amino groups in proteins. The principle is that in the presence of a trace amount of radioactive label, the various reactive groups in a protein molecule will compete for the label and the amount incorporated into any one group will be determined by its nucleophilicity, pK and micro-environment. The relative amounts of label incorporated into various groups will be proportional to their second-order rate constants and by comparing these rate constants with those expected on the basis of a linear free-energy relationship obtained with a series of standard compounds, the micro-environment can be defined for a particular amino group. 2. The method consists of treating a protein and an internal standard with a limiting amount of radioactive reagent and then with an excess of unlabelled reagent to yield a chemically homogeneous but heterogeneously labelled compound. After appropriate enzymic digestion peptides containing each labelled group are isolated and their rates of reaction, relative to the internal standard, are determined from their specific radioactivities. The entire procedure is repeated at several pH values. 3. When the method was applied to the amino groups of porcine elastase by using tritiated acetic anhydride as the labelling reagent, the N-terminus was found to have pKa 9.7 and a much lower than normal reactivity. Lysine-87 and lysine-224 were found to have pKa 10.3 and normal reactivities. At pH values greater than 10.5 there are discontinuities in all the titration curves, indicating that the entire molecule is undergoing a structural reorganization. ImagesPLATE 3PLATE 1PLATE 2 PMID:5158490

  8. Screening and determinations of tissue polypeptide antigen by label-free optical immunosensing method.

    PubMed

    Yao, Zufu; Huang, Kelong; Guo, Jun; Li, Yanhua; Yu, Jingang; Wu, Fangping

    2012-01-01

    Based on the specificity of the immunoreaction of anti-body and antigen, and the resulting localized surface plasmon resonance extinction response of functionalized nano-Au monolayer on glass chip, a novel label-free optical immunosensor with amplified sensitivity has been developed for the detection of tissue polypeptide antigen (TPA). The nano-Au monolayer on glass chip was controllably prepared using 3-mercaptopropyltrimethoxysilane (MPTMOS) as linker by a solution self-assembly method. To analyze its quality, the nano-Au monolayer was characterized by UV-visible spectrum and atomic force microscopy (AFM). The resulting chip was modified by tissue polypeptide antigen antibody (anti-TPA), and then bovine serum albumin (BSA) was employed to block the possible remaining active sites of the nano-Au monolayer. An immunosensor was constructed with biocompatible monolayer nano-Au membrane and the desirable TPA antibody/BSA composite membrane, and it showed good selectivity, high sensitivity and a wide linear response to TPA in the range of 1-1000 ng L(-1) with a detection limit of 1.2 x 10(-3) ng L(-1), as well as good stability and long-term life. Owing to its ease of operation, low detection limit and low cost, it is expected that the proposed procedure may hold great promise in both research-based and clinical assays.

  9. A Simple Method for Labeling Human Embryonic Stem Cells Destined to Lose Undifferentiated Potency

    PubMed Central

    Kumagai, Ayako; Suga, Mika; Yanagihara, Kana; Itoh, Yumi; Furue, Miho K.

    2016-01-01

    Mitochondrial oxidative phosphorylation is a major source of cellular ATP. Its usage as an energy source varies, not only according to the extracellular environment, but also during development and differentiation, as indicated by the reported changes in the flux ratio of glycolysis to oxidative phosphorylation during embryonic stem (ES) cell differentiation. The fluorescent probe JC-1 allows visualization of changes in the mitochondrial membrane potential produced by oxidative phosphorylation. Strong JC-1 signals were localized in the differentiated cells located at the edge of H9 ES colonies that expressed vimentin, an early differentiation maker. The JC-1 signals were further intensified when individual adjacent colonies were in contact with each other. Time-lapse analyses revealed that JC-1-labeled H9 cells under an overconfluent condition were highly differentiated after subculture, suggesting that monitoring oxidative phosphorylation in live cells might facilitate the prediction of induced pluripotent stem cells, as well as ES cells, that are destined to lose their undifferentiated potency. Significance Skillful cell manipulation is a major factor in both maintaining and disrupting the undifferentiation potency of human embryonic stem (hES) cells. Staining with JC-1, a mitochondrial membrane potential probe, is a simple monitoring method that can be used to predict embryonic stem cell quality under live conditions, which might help ensure the future use of hES and human induced pluripotent stem cells after subculture. PMID:26819254

  10. A simple method for N-15 labelling of exocyclic amino groups in synthetic oligodeoxynucleotides

    PubMed Central

    Acedo, Montse; Fàbrega, Carme; Aviño, Anna; Goodman, Myron; Fagan, Patricia; Wemmer, David; Eritja, Ramon

    1994-01-01

    The use of the ammonia deprotection step to introduce 15N labels at specific exocyclic amino positions of adenine, cytosine, guanine or 2-aminopurine of oligodeoxynucleotides is described. PMID:8065910

  11. A rapid and robust method for selective isotope labeling of proteins

    PubMed Central

    Lin, Myat T.; Sperling, Lindsay J.; Frericks Schmidt, Heather L.; Tang, Ming; Samoilova, Rimma I.; Kumasaka, Takashi; Iwasaki, Toshio; Dikanov, Sergei A.; Rienstra, Chad M.; Gennis, Robert B.

    2011-01-01

    Amino-acid selective isotope labeling of proteins offers numerous advantages in mechanistic studies by revealing structural and functional information unattainable from a crystallographic approach. However, efficient labeling of proteins with selected amino acids necessitates auxotrophic hosts, which are often not available. We have constructed a set of auxotrophs in a commonly used Escherichia coli expression strain C43(DE3), a derivative of E. coli BL21(DE3), which can be used for isotopic labeling of individual amino acids or sets of amino acids. These strains have general applicability to either soluble or membrane proteins that can be expressed in E. coli. We present examples in which proteins are selectively labeled with 13C- and 15N-amino acids and studied using magic-angle spinning solid-state NMR and pulsed EPR, demonstrating the utility of these strains for biophysical characterization of membrane proteins, radical-generating enzymes and metalloproteins. PMID:21925267

  12. A rapid and robust method for selective isotope labeling of proteins.

    PubMed

    Lin, Myat T; Sperling, Lindsay J; Frericks Schmidt, Heather L; Tang, Ming; Samoilova, Rimma I; Kumasaka, Takashi; Iwasaki, Toshio; Dikanov, Sergei A; Rienstra, Chad M; Gennis, Robert B

    2011-12-01

    Amino-acid selective isotope labeling of proteins offers numerous advantages in mechanistic studies by revealing structural and functional information unattainable from a crystallographic approach. However, efficient labeling of proteins with selected amino acids necessitates auxotrophic hosts, which are often not available. We have constructed a set of auxotrophs in a commonly used Escherichia coli expression strain C43(DE3), a derivative of E. coli BL21(DE3), which can be used for isotopic labeling of individual amino acids or sets of amino acids. These strains have general applicability to either soluble or membrane proteins that can be expressed in E. coli. We present examples in which proteins are selectively labeled with (13)C- and (15)N-amino acids and studied using magic-angle spinning solid-state NMR and pulsed EPR, demonstrating the utility of these strains for biophysical characterization of membrane proteins, radical-generating enzymes and metalloproteins. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Method and kit for the selective labeling of red blood cells in whole blood with Tc-99m

    DOEpatents

    Srivastava, S.C.; Babich, J.W.; Straub, R.; Richards, P.

    1988-07-05

    Disclosed herein are a method and kit for the preparation of [sup 99m]Tc labeled red blood cells using whole blood in a closed sterile system containing stannous tin in a form such that it will enter the red blood cells and be available for the reduction of technetium. No Drawings

  14. Method and kit for the selective labeling of red blood cells in whole blood with TC-99M

    DOEpatents

    Srivastava, Suresh C.; Babich, John W.; Straub, Rita; Richards, Powell

    1988-01-01

    Disclosed herein are a method and kit for the preparation of .sup.99m Tc labeled red blood cells using whole blood in a closed sterile system containing stannous tin in a form such that it will enter the red blood cells and be available therein for the reduction of technetium.

  15. Methods of sequencing and detection using energy transfer labels with cyanine dyes as donor chromophores

    DOEpatents

    Glazer, Alexander N.; Mathies, Richard A.; Hung, Su-Chun; Ju, Jingyue

    2000-01-01

    Cyanine dyes are used as the donor fluorophore in energy transfer labels in which light energy is absorbed by a donor fluorophore and transferred to an acceptor fluorophore which responds to the transfer by emitting fluorescent light for detection. The cyanine dyes impart an unusually high sensitivity to the labels thereby improving their usefulness in a wide variety of biochemical procedures, particularly nucleic acid sequencing, nucleic acid fragment sizing, and related procedures.

  16. Evaluating Propensity Score Methods in a Quasi-Experimental Study of the Impact of Menu-Labeling

    PubMed Central

    Mayne, Stephanie L.; Lee, Brian K.; Auchincloss, Amy H.

    2015-01-01

    Background Quasi-experimental studies of menu labeling have found mixed results for improving diet. Differences between experimental groups can hinder interpretation. Propensity scores are an increasingly common method to improve covariate balance, but multiple methods exist and the improvements associated with each method have rarely been compared. In this re-analysis of the impact of menu labeling, we compare multiple propensity score methods to determine which methods optimize balance between experimental groups. Methods Study participants included adult customers who visited full-service restaurants with menu labeling (treatment) and without (control). We compared the balance between treatment groups obtained by four propensity score methods: 1) 1:1 nearest neighbor matching (NN), 2) augmented 1:1 NN (using caliper of 0.2 and an exact match on an imbalanced covariate), 3) full matching, and 4) inverse probability weighting (IPW). We then evaluated the treatment effect on differences in nutrients purchased across the different methods. Results 1:1 NN resulted in worse balance than the original unmatched sample (average standardized absolute mean distance [ASAM]: 0.185 compared to 0.171). Augmented 1:1 NN improved balance (ASAM: 0.038) but resulted in a large reduction in sample size. Full matching and IPW improved balance over the unmatched sample without a reduction in sample size (ASAM: 0.049 and 0.031, respectively). Menu labeling was associated with decreased calories, fat, sodium and carbohydrates in the unmatched analysis. Results were qualitatively similar in the propensity score matched/weighted models. Conclusions While propensity scores offer an increasingly popular tool to improve causal inference, choosing the correct method can be challenging. Our results emphasize the benefit of examining multiple methods to ensure results are consistent, and considering approaches beyond the most popular method of 1:1 NN matching. PMID:26677849

  17. Lanthanide-labeled clay: A new method for tracing sediment transport in Karst

    USGS Publications Warehouse

    Mahler, B.J.; Bennett, P.C.; Zimmerman, M.

    1998-01-01

    Mobile sediment is a fundamental yet poorly characterized aspect of mass transport through karst aquifers. Here the development and field testing of an extremely sensitive particle tracer that may be used to characterize sediment transport in karst aquifers is described. The tracer consists of micron-size montmorillonite particles homoionized to the lanthanide form; after injection and retrieval from a ground water system, the lanthanide ions are chemically stripped from the clay and quantified by high performance liquid chromatography. The tracer meets the following desired criteria: low detection limit; a number of differentiable signatures; inexpensive production and quantification using standard methods; no environmental risks; and hydrodynamic properties similar to the in situ sediment it is designed to trace. The tracer was tested in laboratory batch experiments and field tested in both surface water and ground water systems. In surface water, arrival times of the tracer were similar to those of a conservative water tracer, although a significant amount of material was lost due to settling. Two tracer tests were undertaken in a karst aquifer under different flow conditions. Under normal flow conditions, the time of arrival and peak concentration of the tracer were similar to or preceded that of a conservative water tracer. Under low flow conditions, the particle tracer was not detected, suggesting that in low flow the sediment settles out of suspension and goes into storage.Mobile sediment is a fundamental yet poorly characterized aspect of mass transport through karst aquifers. Here the development and field testing of an extremely sensitive particle tracer that may be used to characterize sediment transport in karst aquifers is described. The tracer consists of micron-size montmorillonite particles homoionized to the lanthanide form; after injection and retrieval from a ground water system, the lanthanide ions are chemically stripped from the clay and

  18. Evaluation of a High Intensity Focused Ultrasound-Immobilized Trypsin Digestion and 18 O-Labeling Method for Quantitative Proteomics

    SciTech Connect

    Lopez-Ferrer, Daniel; Hixson, Kim K.; Smallwood, Heather S.; Squier, Thomas C.; Petritis, Konstantinos; Smith, Richard D.

    2009-08-01

    A new method that uses immobilized trypsin concomitant with ultrasonic irradiation results in ultra-rapid digestion and thorough 18O labeling for quantitative protein comparisons. The reproducible and highly efficient method provided effective digestions in <1 min and minimized the amount of enzyme required compared to traditional methods. This method was demonstrated for digestion of both simple and complex protein mixtures, including bovine serum albumin, a global proteome extract from bacteria Shewanella oneidensis, and mouse plasma, as well as for the labeling of complex protein mixtures, which validated the application of this method for differential proteomic measurements. This approach is simple, reproducible, cost effective, and rapid, and thus well-suited for automation.

  19. Tracing nitrogenous disinfection byproducts after medium pressure UV water treatment by stable isotope labeling and high resolution mass spectrometry.

    PubMed

    Kolkman, Annemieke; Martijn, Bram J; Vughs, Dennis; Baken, Kirsten A; van Wezel, Annemarie P

    2015-04-07

    Advanced oxidation processes are important barriers for organic micropollutants (e.g., pharmaceuticals, pesticides) in (drinking) water treatment. Studies indicate that medium pressure (MP) UV/H2O2 treatment leads to a positive response in Ames mutagenicity tests, which is then removed after granulated activated carbon (GAC) filtration. The formed potentially mutagenic substances were hitherto not identified and may result from the reaction of photolysis products of nitrate with (photolysis products of) natural organic material (NOM). In this study we present an innovative approach to trace the formation of disinfection byproducts (DBPs) of MP UV water treatment, based on stable isotope labeled nitrate combined with high resolution mass spectrometry. It was shown that after MP UV treatment of artificial water containing NOM and nitrate, multiple nitrogen containing substances were formed. In total 84 N-DBPs were detected at individual concentrations between 1 to 135 ng/L bentazon-d6 equivalents, with a summed concentration of 1.2 μg/L bentazon-d6 equivalents. The chemical structures of three byproducts were confirmed. Screening for the 84 N-DBPs in water samples from a full-scale drinking water treatment plant based on MP UV/H2O2 treatment showed that 22 of the N-DBPs found in artificial water were also detected in real water samples.

  20. A method for discovering and inferring appropriate eligibility criteria in clinical trial protocols without labeled data

    PubMed Central

    2013-01-01

    Background We consider the user task of designing clinical trial protocols and propose a method that discovers and outputs the most appropriate eligibility criteria from a potentially huge set of candidates. Each document d in our collection D is a clinical trial protocol which itself contains a set of eligibility criteria. Given a small set of sample documentsD′,|D′|≪|D|, a user has initially identified as relevant e.g., via a user query interface, our scoring method automatically suggests eligibility criteria from D, D ⊃ D', by ranking them according to how appropriate they are to the clinical trial protocol currently being designed. The appropriateness is measured by the degree to which they are consistent with the user-supplied sample documents D'. Method We propose a novel three-step method called LDALR which views documents as a mixture of latent topics. First, we infer the latent topics in the sample documents using Latent Dirichlet Allocation (LDA). Next, we use logistic regression models to compute the probability that a given candidate criterion belongs to a particular topic. Lastly, we score each criterion by computing its expected value, the probability-weighted sum of the topic proportions inferred from the set of sample documents. Intuitively, the greater the probability that a candidate criterion belongs to the topics that are dominant in the samples, the higher its expected value or score. Results Our experiments have shown that LDALR is 8 and 9 times better (resp., for inclusion and exclusion criteria) than randomly choosing from a set of candidates obtained from relevant documents. In user simulation experiments using LDALR, we were able to automatically construct eligibility criteria that are on the average 75% and 70% (resp., for inclusion and exclusion criteria) similar to the correct eligibility criteria. Conclusions We have proposed LDALR, a practical method for discovering and inferring appropriate eligibility criteria in clinical

  1. New methods of nitrate removal from water.

    PubMed

    Shrimali, M; Singh, K P

    2001-01-01

    Nitrate contamination in groundwater resources originates mainly from the excessive use of fertilisers and uncontrolled land discharges of treated wastewater. This can cause potential health hazards to infants and pregnant women, thus limiting the direct use of the groundwater resources for the human consumption in several parts of the world, including India. The conventional processes used to eliminate nitrate from water are ion exchange, reverse osmosis and electro-dialysis. The utility of these processes has been limited due to their expensive operation and subsequent disposal problem of the generated nitrate waste brine. This paper presents a comprehensive account of the methods/techniques used for the removal of nitrate ion from water during the last 10 years with special reference to the biological denitrification and fate of the metals in decontamination processes.

  2. Method of arsenic removal from water

    DOEpatents

    Gadgil, Ashok

    2010-10-26

    A method for low-cost arsenic removal from drinking water using chemically prepared bottom ash pre-treated with ferrous sulfate and then sodium hydroxide. Deposits on the surface of particles of bottom ash form of activated iron adsorbent with a high affinity for arsenic. In laboratory tests, a miniscule 5 grams of pre-treated bottom ash was sufficient to remove the arsenic from 2 liters of 2400 ppb (parts per billion) arsenic-laden water to a level below 50 ppb (the present United States Environmental Protection Agency limit). By increasing the amount of pre-treated bottom ash, even lower levels of post-treatment arsenic are expected. It is further expected that this invention supplies a very low-cost solution to arsenic poisoning for large population segments.

  3. Optical methods for quantitative and label-free sensing in living human tissues: principles, techniques, and applications.

    PubMed

    Wilson, Robert H; Vishwanath, Karthik; Mycek, Mary-Ann

    2016-01-01

    We present an overview of quantitative and label-free optical methods used to characterize living biological tissues, with an emphasis on emerging applications in clinical tissue diagnostics. Specifically, this review focuses on diffuse optical spectroscopy, imaging, and tomography, optical coherence-based techniques, and non-linear optical methods for molecular imaging. The potential for non- or minimally-invasive assessment, quantitative diagnostics, and continuous monitoring enabled by these tissue-optics technologies provides significant promise for continued clinical translation.

  4. Water recycling system using thermopervaporation method

    NASA Technical Reports Server (NTRS)

    Nitta, K.; Ashida, A.; Mitani, K.; Ebara, K.; Yamada, A.

    1986-01-01

    A water recycling system concept for the crew of the space station is presented. A thermopervaporation method is a new key technology used for the distillation process, utilizing a hydrophobic membrane. An experimental study of thermopervaporation revealed that the permeation depends on the gap between the membrane and the cooling surface in the condensation room: the steam diffusion occurs with gaps less than 5 mm while natural convection becomes dominant with gaps more than 5 mm. A brief discussion of the system operation is also described.

  5. An isotope-dilution standard GC/MS/MS method for steroid hormones in water

    USGS Publications Warehouse

    Foreman, William T.; Gray, James L.; ReVello, Rhiannon C.; Lindley, Chris E.; Losche, Scott A.

    2013-01-01

    An isotope-dilution quantification method was developed for 20 natural and synthetic steroid hormones and additional compounds in filtered and unfiltered water. Deuterium- or carbon-13-labeled isotope-dilution standards (IDSs) are added to the water sample, which is passed through an octadecylsilyl solid-phase extraction (SPE) disk. Following extract cleanup using Florisil SPE, method compounds are converted to trimethylsilyl derivatives and analyzed by gas chromatography with tandem mass spectrometry. Validation matrices included reagent water, wastewater-affected surface water, and primary (no biological treatment) and secondary wastewater effluent. Overall method recovery for all analytes in these matrices averaged 100%; with overall relative standard deviation of 28%. Mean recoveries of the 20 individual analytes for spiked reagent-water samples prepared along with field samples analyzed in 2009–2010 ranged from 84–104%, with relative standard deviations of 6–36%. Detection levels estimated using ASTM International’s D6091–07 procedure range from 0.4 to 4 ng/L for 17 analytes. Higher censoring levels of 100 ng/L for bisphenol A and 200 ng/L for cholesterol and 3-beta-coprostanol are used to prevent bias and false positives associated with the presence of these analytes in blanks. Absolute method recoveries of the IDSs provide sample-specific performance information and guide data reporting. Careful selection of labeled compounds for use as IDSs is important because both inexact IDS-analyte matches and deuterium label loss affect an IDS’s ability to emulate analyte performance. Six IDS compounds initially tested and applied in this method exhibited deuterium loss and are not used in the final method.

  6. Efficient method for iodine radioisotope labeling of cyclooctyne-containing molecules using strain-promoted copper-free click reaction.

    PubMed

    Jeon, Jongho; Kang, Jung Ae; Shim, Ha Eun; Nam, You Ree; Yoon, Seonhye; Kim, Hye Rim; Lee, Dong Eun; Park, Sang Hyun

    2015-07-01

    Herein we report an efficient method for iodine radioisotope labeling of cyclooctyne-containing molecules using copper-free click reaction. For this study, radioiodination using the tin precursor 2 was carried out at room temperature to give (125)I-labeled azide ([(125)I]1) with high radiochemical yield (85%) and excellent radiochemical purity. Dibenzocyclooctyne (DBCO) containing cRGD peptide and gold nanoparticle were labeled with [(125)I]1 at 37°C for 30min to give triazoles with good radiochemical yields (67-95%). We next carried out tissue biodistribution study of [(125)I]1 in normal ICR mice to investigate the level of organ accumulation which needs to be considered for pre-targeted in vivo imaging. Large amount of [(125)I]1 distributed rapidly in liver and kidney from bloodstream and underwent rapid renal and hepatobiliary clearance. Moreover [(125)I]1 was found to be highly stable (>92%) in mouse serum for 24h. Therefore [(125)I]1 could be used as a potentially useful radiotracer for pre-targeted imaging. Those results clearly indicated that the present radiolabeling method using copper free click reaction would be quite useful for both in vitro and in vivo labeling of DBCO group containing molecules with iodine radioisotopes.

  7. The beauty of being (label)-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics

    PubMed Central

    Campbell, Kate; Deery, Michael J.; Lilley, Kathryn S.; Ralser, Markus

    2014-01-01

    The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA) and data independent (SWATH) acquisition on a TripleTOF 5600 instrument. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides. PMID:24741437

  8. Quantitative estimation of hemorrhage in chronic subdural hematoma using the /sup 51/Cr erythrocyte labeling method

    SciTech Connect

    Ito, H.; Yamamoto, S.; Saito, K.; Ikeda, K.; Hisada, K.

    1987-06-01

    Red cell survival studies using an infusion of chromium-51-labeled erythrocytes were performed to quantitatively estimate hemorrhage in the chronic subdural hematoma cavity of 50 patients. The amount of hemorrhage was determined during craniotomy. Between 6 and 24 hours after infusion of the labeled red cells, hemorrhage accounted for a mean of 6.7% of the hematoma content, indicating continuous or intermittent hemorrhage into the cavity. The clinical state of the patients and the density of the chronic subdural hematoma on computerized tomography scans were related to the amount of hemorrhage. Chronic subdural hematomas with a greater amount of hemorrhage frequently consisted of clots rather than fluid.

  9. Catalytic Enzyme-Based Methods for Water Treatment and Water Distribution System Decontamination. 1. Literature Survey

    DTIC Science & Technology

    2006-06-01

    4. TITLE AND SUBTITLE Sa. CONTRACT NUMBER Catalytic Enzyme-Based Methods for Water Treatment and Water Distribution System Decontamination 1...report was authorized under the "Catalytic Enzyme-Based Methods for Water Treatment and Water Distribution System Decontamination" project funded by the...ENZYME-BASED METHODS FOR WATER TREATMENT AND WATER DISTRIBUTION SYSTEM DECONTAMINATION 1. LITERATURE SURVEY 1. INTRODUCTION Drinking water distribution

  10. Shotgun label-free quantitative proteomics of water-deficit-stressed midmature peanut (Arachis hypogaea L.) seed.

    PubMed

    Kottapalli, Kameswara Rao; Zabet-Moghaddam, Masoud; Rowland, Diane; Faircloth, Wilson; Mirzaei, Mehdi; Haynes, Paul A; Payton, Paxton

    2013-11-01

    Legume seeds and peanuts, in particular, are an inexpensive source of plant proteins and edible oil. A comprehensive understanding of seed metabolism and the effects of water-deficit stress on the incorporation of the main storage reserves in seeds, such as proteins, fatty acids, starch, and secondary metabolites, will enhance our ability to improve seed quality and yield through molecular breeding programs. In the present study, we employed a label-free quantitative proteomics approach to study the functional proteins altered in the midmature (65-70 days postanthesis) peanut seed grown under water-deficit stress conditions. We created a pod-specific proteome database and identified 93 nonredundant, statistically significant, and differentially expressed proteins between well-watered and drought-stressed seeds. Mapping of these differential proteins revealed three candidate biological pathways (glycolysis, sucrose and starch metabolism, and fatty acid metabolism) that were significantly altered due to water-deficit stress. Differential accumulation of proteins from these pathways provides insight into the molecular mechanisms underlying the observed physiological changes, which include reductions in pod yield and biomass, reduced germination, reduced vigor, decreased seed membrane integrity, increase in storage proteins, and decreased total fatty acid content. Some of the proteins encoding rate limiting enzymes of biosynthetic pathways could be utilized by breeders to improve peanut seed production during water-deficit conditions in the field. The data have been deposited to the ProteomeXchange with identifier PXD000308.

  11. A comparison of methods for teaching receptive labeling to children with autism spectrum disorders: a systematic replication.

    PubMed

    Grow, Laura L; Kodak, Tiffany; Carr, James E

    2014-01-01

    Previous research has demonstrated that the conditional-only method (starting with a multiple-stimulus array) is more efficient than the simple-conditional method (progressive incorporation of more stimuli into the array) for teaching receptive labeling to children with autism spectrum disorders (Grow, Carr, Kodak, Jostad, & Kisamore,). The current study systematically replicated the earlier study by comparing the 2 approaches using progressive prompting with 2 boys with autism. The results showed that the conditional-only method was a more efficient and reliable teaching procedure than the simple-conditional method. The results further call into question the practice of teaching simple discriminations to facilitate acquisition of conditional discriminations.

  12. A method for the 32P labeling of peptides or peptide nucleic acid oligomers

    NASA Technical Reports Server (NTRS)

    Kozlov, I. A.; Nielsen, P. E.; Orgel, L. E.; Bada, J. L. (Principal Investigator)

    1998-01-01

    A novel approach to the radioactive labeling of peptides and PNA oligomers is described. It is based on the conjugation of a deoxynucleoside 3'-phosphate with the terminal amine of the substrate, followed by phosphorylation of the 5'-hydroxyl group of the nucleotide using T4 polynucleotide kinase and [gamma-32P]ATP.

  13. A Method for Label-Free, Differential Top-Down Proteomics

    PubMed Central

    Ntai, Ioanna; Toby, Timothy K.; LeDuc, Richard D.; Kelleher, Neil L.

    2016-01-01

    Biomarker discovery in the translational research has heavily relied on labeled and label-free quantitative bottom-up proteomics. Here, we describe a new approach to biomarker studies that utilizes high-throughput top-down proteomics and is the first to offer whole protein characterization and relative quantitation within the same experiment. Using yeast as a model, we report procedures for a label-free approach to quantify the relative abundance of intact proteins ranging from 0 to 30 kDa in two different states. In this chapter, we describe the integrated methodology for the large-scale profiling and quantitation of the intact proteome by liquid chromatography-mass spectrometry (LC-MS) without the need for metabolic or chemical labeling. This recent advance for quantitative top-down proteomics is best implemented with a robust and highly controlled sample preparation workflow before data acquisition on a high-resolution mass spectrometer, and the application of a hierarchical linear statistical model to account for the multiple levels of variance contained in quantitative proteomic comparisons of samples for basic and clinical research. PMID:26867742

  14. A Method for Label-Free, Differential Top-Down Proteomics.

    PubMed

    Ntai, Ioanna; Toby, Timothy K; LeDuc, Richard D; Kelleher, Neil L

    2016-01-01

    Biomarker discovery in the translational research has heavily relied on labeled and label-free quantitative bottom-up proteomics. Here, we describe a new approach to biomarker studies that utilizes high-throughput top-down proteomics and is the first to offer whole protein characterization and relative quantitation within the same experiment. Using yeast as a model, we report procedures for a label-free approach to quantify the relative abundance of intact proteins ranging from 0 to 30 kDa in two different states. In this chapter, we describe the integrated methodology for the large-scale profiling and quantitation of the intact proteome by liquid chromatography-mass spectrometry (LC-MS) without the need for metabolic or chemical labeling. This recent advance for quantitative top-down proteomics is best implemented with a robust and highly controlled sample preparation workflow before data acquisition on a high-resolution mass spectrometer, and the application of a hierarchical linear statistical model to account for the multiple levels of variance contained in quantitative proteomic comparisons of samples for basic and clinical research.

  15. A simple method to determine mineralization of (14) C-labeled compounds in soil.

    PubMed

    Myung, Kyung; Madary, Michael W; Satchivi, Norbert M

    2014-06-01

    Degradation of organic compounds in soil is often determined by measuring the decrease of the parent compound and analyzing the occurrence of its metabolites. However, determining carbon species as end products of parent compound dissipation requires using labeled materials that allow more accurate determination of the environmental fate of the compound of interest. The current conventional closed system widely used to monitor degradation of (14) C-labeled compounds in soil is complex and expensive and requires a specialized apparatus and facility. In the present study, the authors describe a simple system that facilitates measurement of mineralization of (14) C-labeled compounds applied to soil samples. In the system, soda lime pellets to trap mineralized (14) C-carbon species, including carbon dioxide, were placed in a cup, which was then inserted above the treated soil sample in a tube. Mineralization of [(14) C]2,4-D applied to soil samples in the simple system was compared with that in the conventional system. The simple system provided an equivalent detection of (14) C-carbon species mineralized from the parent compound. The results demonstrate that this cost- and space-effective simple system is suitable for examining degradation and mineralization of (14) C-labeled compounds in soil and could potentially be used to investigate their mineralization in other biological matrices. © 2014 SETAC.

  16. A simple method for in vivo labelling of infiltrating leukocytes in the mouse retina using indocyanine green dye

    PubMed Central

    Sim, Dawn A.; Chu, Colin J.; Selvam, Senthil; Powner, Michael B.; Liyanage, Sidath; Copland, David A.; Keane, Pearse A.; Tufail, Adnan; Egan, Catherine A.; Bainbridge, James W. B.; Lee, Richard W.; Dick, Andrew D.; Fruttiger, Marcus

    2015-01-01

    ABSTRACT We have developed a method to label and image myeloid cells infiltrating the mouse retina and choroid in vivo, using a single depot injection of indocyanine green dye (ICG). This was demonstrated using the following ocular models of inflammation and angiogenesis: endotoxin-induced uveitis, experimental autoimmune uveoretinitis and laser-induced choroidal neovascularization model. A near-infrared scanning ophthalmoscope was used for in vivo imaging of the eye, and flow cytometry was used on blood and spleen to assess the number and phenotype of labelled cells. ICG was administered 72 h before the induction of inflammation to ensure clearance from the systemic circulation. We found that in vivo intravenous administration failed to label any leukocytes, whereas depot injection, either intraperitoneal or subcutaneous, was successful in labelling leukocytes infiltrating into the retina. Progression of inflammation in the retina could be traced over a period of 14 days following a single depot injection of ICG. Additionally, bright-field microscopy, spectrophotometry and flow cytometric analysis suggest that the predominant population of cells stained by ICG are circulating myeloid cells. The translation of this approach into clinical practice would enable visualization of immune cells in situ. This will not only provide a greater understanding of pathogenesis, monitoring and assessment of therapy in many human ocular diseases but might also open the ability to image immunity live for neurodegenerative disorders, cardiovascular disease and systemic immune-mediated disorders. PMID:26398933

  17. A simple method for in vivo labelling of infiltrating leukocytes in the mouse retina using indocyanine green dye.

    PubMed

    Sim, Dawn A; Chu, Colin J; Selvam, Senthil; Powner, Michael B; Liyanage, Sidath; Copland, David A; Keane, Pearse A; Tufail, Adnan; Egan, Catherine A; Bainbridge, James W B; Lee, Richard W; Dick, Andrew D; Fruttiger, Marcus

    2015-11-01

    We have developed a method to label and image myeloid cells infiltrating the mouse retina and choroid in vivo, using a single depot injection of indocyanine green dye (ICG). This was demonstrated using the following ocular models of inflammation and angiogenesis: endotoxin-induced uveitis, experimental autoimmune uveoretinitis and laser-induced choroidal neovascularization model. A near-infrared scanning ophthalmoscope was used for in vivo imaging of the eye, and flow cytometry was used on blood and spleen to assess the number and phenotype of labelled cells. ICG was administered 72 h before the induction of inflammation to ensure clearance from the systemic circulation. We found that in vivo intravenous administration failed to label any leukocytes, whereas depot injection, either intraperitoneal or subcutaneous, was successful in labelling leukocytes infiltrating into the retina. Progression of inflammation in the retina could be traced over a period of 14 days following a single depot injection of ICG. Additionally, bright-field microscopy, spectrophotometry and flow cytometric analysis suggest that the predominant population of cells stained by ICG are circulating myeloid cells. The translation of this approach into clinical practice would enable visualization of immune cells in situ. This will not only provide a greater understanding of pathogenesis, monitoring and assessment of therapy in many human ocular diseases but might also open the ability to image immunity live for neurodegenerative disorders, cardiovascular disease and systemic immune-mediated disorders. © 2015. Published by The Company of Biologists Ltd.

  18. Measurement of Energy Expenditure under Field Conditions Using Doubly Labeled Water

    DTIC Science & Technology

    1988-08-31

    nstitutes of Health . j ’PI Signature D ’ 1m TABLE OF CONTENTS Page IWTROUCTION 3 I Hypothesis 3 II Objectives 3 III Military Significance 3 VI Background 3...Surface water for use as drinking water was abundant during this field training exercise. The study was conducted in late September and October, with...8used to correct isotope abundances in subjects receiving H 0. These changes are quite close to the anticipated change. Although actual drinking water

  19. The water method combined with chromoendoscopy enhances adenoma detection

    PubMed Central

    Ransibrahmanakul, Kanat; Toomsen, Lee; Mann, Surinder K; Siao-Salera, Rodelei

    2011-01-01

    Background The water method is easy-to-learn and improves colonoscopy outcomes. Dye-spray chromoendoscopy enhances ADR but has not been widely accepted for routine application in screening or surveillance colonoscopy. Hypothesis With dye added to the water used in the water method, ADR can be enhanced compared with the water or air method alone. Objective To compare ADR determined by the air method, water method alone, and water method with indigo carmine (0.008%) added. Design Review of prospectively collected data in a performance improvement program. Setting VA endoscopy unit. Patient Screening or surveillance colonoscopy. Methods Patients (n=50/group) underwent colonoscopy with each of the three methods. Water method involved warm water infusion in lieu of air insufflation coupled with removal of residual air by suction and residual feces by water exchange. ADR and procedural data were collected prospectively to monitor performance. Main Outcome Measurements ADR. Results ADR in the air method, water method alone and water method with indigo carmine were 36%, 40% and 62%, respectively. Water method with indigo carmine produced significantly higher ADR than the air or water method alone (p<0.05). Limitations Non-randomized data, single VA site, retrospective comparison. Absence of significant difference between air and water methods could be a type II error due to small number of patients Conclusions The approach with indigo carmine added to the water used in the water method yielded significantly higher ADR than the water or the air method alone. The data suggest that a prospective RCT to compare the different methods is warranted. PMID:21776426

  20. Membrane filter-fluorescent-antibody method for detection and enumeration of bacteria in water.

    PubMed

    Guthrie, R K; Reeder, D J

    1969-03-01

    A technique which employs nonfluorescing membrane filters and specific fluoresceinisothiocynate-labeled antiserum has been successfully used in the identification and enumeration of known species of Escherichia coli which have been added to natural populations of bacteria found in water. The quantitative results compared favorably with those of standard tests. The use of a dissecting microscope with an external lighting arrangement provided a simple requirement for equipment. This method may be useful in monitoring specific bacterial types from waters which were being monitored for specific pollution.

  1. A new automated NaCl based robust method for routine production of gallium-68 labeled peptides

    PubMed Central

    Schultz, Michael K.; Mueller, Dirk; Baum, Richard P.; Watkins, G. Leonard; Breeman, Wouter A. P.

    2017-01-01

    A new NaCl based method for preparation of gallium-68 labeled radiopharmaceuticals has been adapted for use with an automated gallium-68 generator system. The method was evaluated based on 56 preparations of [68Ga]DOTATOC and compared to a similar acetone-based approach. Advantages of the new NaCl approach include reduced preparation time (< 15 min) and removal of organic solvents. The method produces high peptide-bound % (> 97%), and specific activity (> 40 MBq nmole−1 [68Ga]DOTATOC) and is well-suited for clinical production of radiopharmaceuticals. PMID:23026223

  2. A new automated NaCl based robust method for routine production of gallium-68 labeled peptides.

    PubMed

    Schultz, Michael K; Mueller, Dirk; Baum, Richard P; Leonard Watkins, G; Breeman, Wouter A P

    2013-06-01

    A new NaCl based method for preparation of gallium-68 labeled radiopharmaceuticals has been adapted for use with an automated gallium-68 generator system. The method was evaluated based on 56 preparations of [(68)Ga]DOTATOC and compared to a similar acetone-based approach. Advantages of the new NaCl approach include reduced preparation time (<15 min) and removal of organic solvents. The method produces high peptide-bound % (>97%), and specific activity (>40 MBq nmole(-1) [(68)Ga]DOTATOC) and is well-suited for clinical production of radiopharmaceuticals. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. All-optical recognition method of double two-dimensional optical orthogonal codes-based labels using four-wave mixing

    NASA Astrophysics Data System (ADS)

    Zhang, Chongfu; Wang, Leyang; Perumal, Sathishkumar; Qiu, Kun; Zhou, Heng

    2011-08-01

    A novel all-optical label recognition method is proposed and demonstrated experimentally which is based on fiber Bragg gratings (FBGs)-based encoder/decoder and semiconductor optical amplifier (SOA). In this scheme, the optical label is firstly decoded properly, the decoded signal then generates the 1st and the 2nd order four-wave mixing (FWM) effect in different SOA, any of the frequencies achieved by the 2nd order FWM is extracted to recognize the optical label. The proposed solution can favor hardware simplicity over bandwidth efficiency in order to achieve the double two-dimensional optical orthogonal codes (2D-OOCs)-based optical label recognition in an optical packet switching (OPS) system where the bandwidth efficiency can be improved by FWM effect in SOA to achieve optical label processing and reasonable spacing of wavelengths for the payloads and optical label. The feasibility of the proposed method is validated by two experiments of the double 2D-OOCs-based optical label generation and recognition, the effect of the optical label on the payloads is also considered. These results show that the proposed method can (1) reduce effectively the code auto-correlation /cross-correlation requirements of the optical label identification and remove the cross-correlation pulses after optical decoding, (2) increase greatly the coding capacity and the number of the available optical labels, (3) improve the reliability and bandwidth efficiency of the optical label identification. The experimental results also show that the optical label has a high extinction ratio and can be operated easily.

  4. A new metabolic cell-wall labelling method reveals peptidoglycan in Chlamydia trachomatis.

    PubMed

    Liechti, G W; Kuru, E; Hall, E; Kalinda, A; Brun, Y V; VanNieuwenhze, M; Maurelli, A T

    2014-02-27

    Peptidoglycan (PG), an essential structure in the cell walls of the vast majority of bacteria, is critical for division and maintaining cell shape and hydrostatic pressure. Bacteria comprising the Chlamydiales were thought to be one of the few exceptions. Chlamydia harbour genes for PG biosynthesis and exhibit susceptibility to 'anti-PG' antibiotics, yet attempts to detect PG in any chlamydial species have proven unsuccessful (the 'chlamydial anomaly'). We used a novel approach to metabolically label chlamydial PG using d-amino acid dipeptide probes and click chemistry. Replicating Chlamydia trachomatis were labelled with these probes throughout their biphasic developmental life cycle, and the results of differential probe incorporation experiments conducted in the presence of ampicillin are consistent with the presence of chlamydial PG-modifying enzymes. These findings culminate 50 years of speculation and debate concerning the chlamydial anomaly and are the strongest evidence so far that chlamydial species possess functional PG.

  5. New Method for Producing Significant Amounts of RNA Labeled at Specific Sites | Center for Cancer Research

    Cancer.gov

    Among biomacromolecules, RNA is the most versatile, and it plays indispensable roles in almost all aspects of biology. For example, in addition to serving as mRNAs coding for proteins, RNAs regulate gene expression, such as controlling where, when, and how efficiently a gene gets expressed, participate in RNA processing, encode the genetic information of some viruses, serve as scaffolds, and even possess enzymatic activity. To study these RNAs and their biological functions and to make use of those RNA activities for biomedical applications, researchers first need to make various types of RNA. For structural biologists incorporating modified or labeled nucleotides at specific sites in RNA molecules of interest is critical to gain structural insight into RNA functions. However, placing labeled or modified residue(s) in desired positions in a large RNA has not been possible until now.

  6. Carboxyfluorescein diacetate succinimidyl ester labeling method to study the interaction between Leptospira and macrophages.

    PubMed

    Liu, Boyu; Wang, Yanchun; Guo, Xiaokui; Zhu, Weinan; Zhang, Yan; He, Ping

    2014-12-01

    Leptospirosis, which is caused by pathogenic species of the genus Leptospira, has emerged as one of the most widespread zoonotic diseases in the world. The exact mechanism of pathogenesis remains unknown, and the interaction between Leptospira and macrophages is not well understood. In this study, we report that carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) can efficiently label different Leptospira interrogans strains without affecting bacterial motility, viability, or virulence. Following co-incubation, CFDA-SE-labeled leptospires associated with macrophages were quantified by flow cytometry or confocal microscopy. In addition, we showed that trypan blue efficiently quenched the extracellular fluorescence from the adherent leptospires, which enabled intracellular and extracellular bacteria to be distinguished. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Anterograde Tracing Method using DiI to Label Vagal Innervation of the Embryonic and Early Postnatal Mouse Gastrointestinal Tract

    PubMed Central

    Murphy, Michelle C.; Fox, Edward A.

    2007-01-01

    The mouse is an extremely valuable model for studying vagal development in relation to strain differences, genetic variation, gene manipulations, or pharmacological manipulations. Therefore, a method using 1, 1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) was developed for labeling vagal innervation of the gastrointestinal (GI) tract in embryonic and postnatal mice. DiI labeling was adapted and optimized for this purpose by varying several facets of the method. For example, insertion and crushing of DiI crystals into the nerve led to faster DiI diffusion along vagal axons and diffusion over longer distances as compared with piercing the nerve with a micropipette tip coated with dried DiI oil. Moreover, inclusion of EDTA in the fixative reduced leakage of DiI out of nerve fibers that occurred with long incubations. Also, mounting labeled tissue in PBS was superior to glycerol with n-propyl gallate, which resulted in reduced clarity of DiI labeling that may have been due to DiI leaking out of fibers. Optical sectioning of flattened wholemounts permitted examination of individual tissue layers of the GI tract wall. This procedure aided identification of nerve ending types because in most instances each type innervates a different tissue layer. Between embryonic day 12.5 and postnatal day 8, growth of axons into the GI tract, formation and patterning of fiber bundles in the myenteric plexus and early formation of putative afferent and efferent nerve terminals were observed. Thus, the DiI tracing method developed here has opened up a window for investigation during an important phase of vagal development. PMID:17418900

  8. Method for preparing radionuclide-labeled chelating agent-ligand complexes

    DOEpatents

    Meares, Claude F.; Li, Min; DeNardo, Sally J.

    1999-01-01

    Radionuclide-labeled chelating agent-ligand complexes that are useful in medical diagnosis or therapy are prepared by reacting a radionuclide, such as .sup.90 Y or .sup.111 In, with a polyfunctional chelating agent to form a radionuclide chelate that is electrically neutral; purifying the chelate by anion exchange chromatography; and reacting the purified chelate with a targeting molecule, such as a monoclonal antibody, to form the complex.

  9. Methods for Environments and Contaminants: Drinking Water

    EPA Pesticide Factsheets

    EPA’s Safe Drinking Water Information System Federal Version (SDWIS/FED) includes information on populations served and violations of maximum contaminant levels or required treatment techniques by the nation’s 160,000 public water systems.

  10. Deep Label Distribution Learning With Label Ambiguity

    NASA Astrophysics Data System (ADS)

    Gao, Bin-Bin; Xing, Chao; Xie, Chen-Wei; Wu, Jianxin; Geng, Xin

    2017-06-01

    Convolutional Neural Networks (ConvNets) have achieved excellent recognition performance in various visual recognition tasks. A large labeled training set is one of the most important factors for its success. However, it is difficult to collect sufficient training images with precise labels in some domains such as apparent age estimation, head pose estimation, multi-label classification and semantic segmentation. Fortunately, there is ambiguous information among labels, which makes these tasks different from traditional classification. Based on this observation, we convert the label of each image into a discrete label distribution, and learn the label distribution by minimizing a Kullback-Leibler divergence between the predicted and ground-truth label distributions using deep ConvNets. The proposed DLDL (Deep Label Distribution Learning) method effectively utilizes the label ambiguity in both feature learning and classifier learning, which help prevent the network from over-fitting even when the training set is small. Experimental results show that the proposed approach produces significantly better results than state-of-the-art methods for age estimation and head pose estimation. At the same time, it also improves recognition performance for multi-label classification and semantic segmentation tasks.

  11. Biomonitoring Methods for Drinking Water Protection

    DTIC Science & Technology

    2004-12-01

    aquatic biomonitor continuously monitors water at fixed facilities such as water treatment plants by identifying changes in fish ventilatory and...for use in rear areas and at fixed Army facilities such as water treatment plants . To decrease the size, weight, and logistic requirements of the...conjunction with one of the two sets of eight fish in the biomonitor at the Fort Detrick water treatment plant ; the other set of eight fish was exposed

  12. Methods for virus recovery in water

    USDA-ARS?s Scientific Manuscript database

    Food safety is intimately connected to water sanitary quality as water is used at almost every node in the food production process. Common contaminating pathogens in water are human enteric viruses, many of which are responsible for foodborne disease outbreaks in the United States and other high-inc...

  13. Label-free high-throughput screening via mass spectrometry: a single cystathionine quantitative method for multiple applications.

    PubMed

    Holt, Tom G; Choi, Bernard K; Geoghagen, Neil S; Jensen, Kristian K; Luo, Qi; LaMarr, William A; Makara, Gergely M; Malkowitz, Lorraine; Ozbal, Can C; Xiong, Yusheng; Dufresne, Claude; Luo, Ming-Juan

    2009-10-01

    Label-free mass spectrometric (MS) technologies are particularly useful for enzyme assay design for drug discovery screens. MS permits the selective detection of enzyme substrates or products in a wide range of biological matrices without need for derivatization, labeling, or capture technologies. As part of a cardiovascular drug discovery effort aimed at finding modulators of cystathionine beta-synthase (CBS), we used the RapidFire((R)) label-free high-throughput MS (HTMS) technology to develop a high-throughput screening (HTS) assay for CBS activity. The in vitro assay used HTMS to quantify the unlabeled product of the CBS reaction, cystathionine. Cystathionine HTMS analyses were carried out with a throughput of 7 s per sample and quantitation over a linear range of 80-10,000 nM. A compound library of 25,559 samples (or 80 384-well plates) was screened as singlets using the HTMS assay in a period of 8 days. With a hit rate of 0.32%, the actives showed a 90% confirmation rate. The in vitro assay was applied to secondary screens in more complex matrices with no additional analytical development. Our results show that the HTMS method was useful for screening samples containing serum, for cell-based assays, and for liver explants. The novel extension of the in vitro analytical method, without modification, to secondary assays resulted in a significant and advantageous economy of development time for the drug discovery project.

  14. Methods for labeling skeletal muscle ion channels site-specifically with fluorophores suitable for FRET-based structural analysis.

    PubMed

    Mahalingam, Mohana; Fessenden, James D

    2015-01-01

    Skeletal muscle excitation-contraction coupling is triggered by the concerted action of two enormous Ca(2+) channel complexes, the dihydropyridine receptor and the type 1 ryanodine receptor. Recent advances in our understanding of the structure of these large Ca(2+) channels have been driven by fluorescence resonance energy transfer (FRET)-based analysis. A methodological challenge in conducting these FRET measurements is the ability to site-specifically label these huge ion channels with donor and acceptor fluorophores capable of undergoing energy transfer. In this chapter, we detail specific protocols for tagging large membrane proteins with these fluorescent probes using three orthogonal labeling methods: fluorescent protein fusions, biarsenical reagents directed to engineered tetracysteine tags, and Cy3/5 nitrilotriacetic acid conjugates that bind to poly-histidine tags. © 2015 Elsevier Inc. All rights reserved.

  15. Sparse Markov chain-based semi-supervised multi-instance multi-label method for protein function prediction.

    PubMed

    Han, Chao; Chen, Jian; Wu, Qingyao; Mu, Shuai; Min, Huaqing

    2015-10-01

    Automated assignment of protein function has received considerable attention in recent years for genome-wide study. With the rapid accumulation of genome sequencing data produced by high-throughput experimental techniques, the process of manually predicting functional properties of proteins has become increasingly cumbersome. Such large genomics data sets can only be annotated computationally. However, automated assignment of functions to unknown protein is challenging due to its inherent difficulty and complexity. Previous studies have revealed that solving problems involving complicated objects with multiple semantic meanings using the multi-instance multi-label (MIML) framework is effective. For the protein function prediction problems, each protein object in nature may associate with distinct structural units (instances) and multiple functional properties (class labels) where each unit is described by an instance and each functional property is considered as a class label. Thus, it is convenient and natural to tackle the protein function prediction problem by using the MIML framework. In this paper, we propose a sparse Markov chain-based semi-supervised MIML method, called Sparse-Markov. A sparse transductive probability graph is constructed to encode the affinity information of the data based on ensemble of Hausdorff distance metrics. Our goal is to exploit the affinity between protein objects in the sparse transductive probability graph to seek a sparse steady state probability of the Markov chain model to do protein function prediction, such that two proteins are given similar functional labels if they are close to each other in terms of an ensemble Hausdorff distance in the graph. Experimental results on seven real-world organism data sets covering three biological domains show that our proposed Sparse-Markov method is able to achieve better performance than four state-of-the-art MIML learning algorithms.

  16. Heavy water and 15N labeling with NanoSIMS analysis reveals growth-rate dependent metabolic heterogeneity in chemostats

    PubMed Central

    McGlynn, Shawn E.; Green-Saxena, Abigail

    2015-01-01

    To measure single cell microbial activity and substrate utilization patterns in environmental systems, we employ a new technique using stable isotope labeling of microbial populations with heavy water (a passive tracer) and 15N ammonium in combination with multi-isotope imaging mass spectrometry. We demonstrate simultaneous NanoSIMS analysis of hydrogen, carbon and nitrogen at high spatial and mass resolution, and report calibration data linking single cell isotopic compositions to the corresponding bulk isotopic equivalents for Pseudomonas aeruginosa and Staphylococcus aureus. Our results show that heavy water is capable of quantifying in situ single cell microbial activities ranging from generational time scales of minutes to years, with only light isotopic incorporation (∼0.1 atom % 2H). Applying this approach to study the rates of fatty acid biosynthesis by single cells of S. aureus growing at different rates in chemostat culture (∼6 hours, 1 day and 2 week generation times), we observe the greatest anabolic activity diversity in the slowest growing populations. By using heavy water to constrain cellular growth activity, we can further infer the relative contributions of ammonium vs. amino acid assimilation to the cellular nitrogen pool. The approach described here can be applied to disentangle individual cell activities even in nutritionally complex environments. PMID:25655651

  17. A method for double-labeling sputum cells for p53 and cytokeratin

    SciTech Connect

    Neft, R.E.; Tierney, L.A.; Belinsky, S.A.

    1995-12-01

    Molecular and immunological techniques may enhance the usefulness of sputum cytology as a screening tool for lung cancer. These techniques may also be useful in detecting and following the early progression of disease from metaplasia to dysplasia, carcinoma in situ, and finally to invasive carcinoma. Longitudinal information on the evolution of these malignant changes in the respiratory epithelium can be gained by prospective study of populations at high risk for lung cancer. This work is significant because double-labeling of cells in sputum with p53 and cytokeratin antibodies facilitates rapid screening of p53 positive neoplastic and preneoplastic lung cells by brightfield and fluorescence microscopy.

  18. Microneedle Biosensor: A Method for Direct Label-free Real Time Protein Detection

    PubMed Central

    Esfandyarpour, Rahim; Esfandyarpour, Hesaam; Javanmard, Mehdi; Harris, James S.; Davis, Ronald W.

    2012-01-01

    Here we present the development of an array of electrical micro-biosensors in a microfluidic channel, called microneedle biosensors. A microneedle biosensor is a real-time, label-free, direct electrical detection platform, which is capable of high sensitivity detection, measuring the change in ionic current and impedance modulation, due to the presence or reaction of biomolecules such as proteins and nucleic acids. In this study, we successfully fabricated and electrically characterized the sensors and demonstrated successful detection of target protein. In this study, we used biotinylated bovine serum albumin as the receptor and streptavidin as the target analyte PMID:23355762

  19. Household adoption of energy and water-efficient appliances: An analysis of attitudes, labelling and complementary green behaviours in selected OECD countries.

    PubMed

    Dieu-Hang, To; Grafton, R Quentin; Martínez-Espiñeira, Roberto; Garcia-Valiñas, Maria

    2017-03-28

    Using a household-based data set of more than 12,000 households from 11 OECD countries, we analyse the factors underlying the decision by households to adopt energy-efficient and water-efficient equipment. We evaluate the roles of both attitudes and labelling schemes on the adoption of energy and water-efficient equipment, and also the interaction and complementarity between energy and water conservation behaviours. Our findings show: one, 'green' social norms and favourable attitudes towards the environment are associated with an increased likelihood of households' adoption of energy and water-efficient appliances; two, households' purchase decisions are positively affected by their awareness, understanding, and trust of labelling schemes; and three, there is evidence of complementarity between energy conservation and water conservation behaviours.

  20. Use of isotopically labeled fertilizer to trace nitrogen fertilizer contributions to surface, soil, and ground water

    USGS Publications Warehouse

    Wilkison, D.H.; Blevins, D.W.; Silva, S.R.

    2000-01-01

    The fate and transport of a single N fertilizer application through plants, soil, runoff, and the unsaturated and saturated zones was determined for four years at a field site under continuous corn (Zea mays L.) management. Claypan soils, which underlie the site, were hypothesized to restrict the movement of agrichemicals from the soil surface to ground water. However, N fertilizer moved rapidly through preferential flow paths in the soil and into the underlying glacial till aquifer. Most N transport occurred during the fall and winter when crops were not available to use excess N. Forty months after application, 33 percent of the fertilizer had been removed by grain harvests, 30 percent had been transpired to the atmosphere, and 33 percent had migrated to ground water. Although runoff volumes were 50 percent greater than infiltration, less than 2 percent of the fertilizer was lost to runoff. Small measured denitrification rates and large measured dissolved oxygen concentrations in ground water favor the long-term stability of NO3-1 in ground water. Successive fertilizer applications, in areas that lack the ability to moderate N concentrations through consumptive N reactions, risk the potential of N-saturated ecosystems.

  1. Study methods for disinfection water for injection

    NASA Astrophysics Data System (ADS)

    Grishkanich, Alexander; Zhevlakov, Alexander; Kascheev, Sergey; Polyakov, Vladimir; Sidorov, Igor; Ruzankina, Julia; Yakovlev, Alexey; Mak, Andrey

    2016-04-01

    Experimental results presented in this study tends to explore viruses in the water for their further decontamination under the influence of laser radiation (λ=220-390 nm). Conducted a series of experiments to study the dependence of water quality from the effects of laser radiation. Correlation between degree of survival of viruses and power density. The results showed that all the analyzed samples of water is clearing from bacteria to 98%. Preliminary tests of the prototype laboratory system UFOVI has opened up new opportunities for water sterilizing.

  2. Electrophoretic method for the determination of the proportion of gamma-aminobutyric acid in a mixture of labeled neurotransmitter and its catabolites

    SciTech Connect

    Cupello, A.; Rapallino, M.V.; Besio, G.; Mainardi, P.

    1987-01-01

    An electrophoretic method for the separation of gamma-aminobutyric acid (GABA) from its metabolites after GABA-transaminase attack is presented. The method is based on the fact that at neutral pH GABA has no net electrical charge, whereas its major metabolites, succinic acid and Krebs cycle intermediates, are negatively charged. The method appears to be especially suitable for evaluation of true-labeled neurotransmitter within the radioactivity which is found in synaptosomes after labeled GABA-uptake studies.

  3. Direct methods and residue type specific isotope labeling in NMR structure determination and model-driven sequential assignment.

    PubMed

    Schedlbauer, Andreas; Auer, Renate; Ledolter, Karin; Tollinger, Martin; Kloiber, Karin; Lichtenecker, Roman; Ruedisser, Simon; Hommel, Ulrich; Schmid, Walther; Konrat, Robert; Kontaxis, Georg

    2008-10-01

    Direct methods in NMR based structure determination start from an unassigned ensemble of unconnected gaseous hydrogen atoms. Under favorable conditions they can produce low resolution structures of proteins. Usually a prohibitively large number of NOEs is required, to solve a protein structure ab-initio, but even with a much smaller set of distance restraints low resolution models can be obtained which resemble a protein fold. One problem is that at such low resolution and in the absence of a force field it is impossible to distinguish the correct protein fold from its mirror image. In a hybrid approach these ambiguous models have the potential to aid in the process of sequential backbone chemical shift assignment when (13)C(beta) and (13)C' shifts are not available for sensitivity reasons. Regardless of the overall fold they enhance the information content of the NOE spectra. These, combined with residue specific labeling and minimal triple-resonance data using (13)C(alpha) connectivity can provide almost complete sequential assignment. Strategies for residue type specific labeling with customized isotope labeling patterns are of great advantage in this context. Furthermore, this approach is to some extent error-tolerant with respect to data incompleteness, limited precision of the peak picking, and structural errors caused by misassignment of NOEs.

  4. A Quick and Parallel Analytical Method Based on Quantum Dots Labeling for ToRCH-Related Antibodies

    NASA Astrophysics Data System (ADS)

    Yang, Hao; Guo, Qing; He, Rong; Li, Ding; Zhang, Xueqing; Bao, Chenchen; Hu, Hengyao; Cui, Daxiang

    2009-12-01

    Quantum dot is a special kind of nanomaterial composed of periodic groups of II-VI, III-V or IV-VI materials. Their high quantum yield, broad absorption with narrow photoluminescence spectra and high resistance to photobleaching, make them become a promising labeling substance in biological analysis. Here, we report a quick and parallel analytical method based on quantum dots for ToRCH-related antibodies including Toxoplasma gondii, Rubella virus, Cytomegalovirus and Herpes simplex virus type 1 (HSV1) and 2 (HSV2). Firstly, we fabricated the microarrays with the five kinds of ToRCH-related antigens and used CdTe quantum dots to label secondary antibody and then analyzed 100 specimens of randomly selected clinical sera from obstetric outpatients. The currently prevalent enzyme-linked immunosorbent assay (ELISA) kits were considered as “golden standard” for comparison. The results show that the quantum dots labeling-based ToRCH microarrays have comparable sensitivity and specificity with ELISA. Besides, the microarrays hold distinct advantages over ELISA test format in detection time, cost, operation and signal stability. Validated by the clinical assay, our quantum dots-based ToRCH microarrays have great potential in the detection of ToRCH-related pathogens.

  5. Toward a Noninvasive, Label-Free Screening Method for Determining Spore Inoculum Quality of Penicillium chrysogenum Using Raman Spectroscopy.

    PubMed

    Wieland, Karin; Kuligowski, Julia; Ehgartner, Daniela; Ramer, Georg; Koch, Cosima; Ofner, Johannes; Herwig, Christoph; Lendl, Bernhard

    2017-01-01

    We report on a label-free, noninvasive method for determination of spore inoculum quality of Penicillium chrysogenum prior to cultivation/germination. Raman microspectroscopy providing direct, molecule-specific information was used to extract information on the viability state of spores sampled directly from the spore inoculum. Based on the recorded Raman spectra, a supervised classification method was established for classification between living and dead spores and thus determining spore inoculum quality for optimized process control. A fast and simple sample preparation method consisting of one single dilution step was employed to eliminate interfering signals from the matrix and to achieve isolation of single spores on the sample carrier (CaF2). Aiming to avoid any influence of the killing procedure in the Raman spectrum of the spore, spores were considered naturally dead after more than one year of storage time. Fluorescence staining was used as reference method. A partial least squares discriminant analysis classifier was trained with Raman spectra of 258 living and dead spores (178 spectra for calibration, 80 spectra for validation). The classifier showed good performance when being applied to a 1 µL droplet taken from a 1:1 mixture of living and dead spores. Of 135 recorded spectra, 51% were assigned to living spores while 49% were identified as dead spores by the classifier. The results obtained in this work are a fundamental step towards developing an automated, label-free, and noninvasive screening method for assessing spore inoculum quality.

  6. Label-free detection of zinc oxide nanowire using a graphene wrapping method.

    PubMed

    You, Juneseok; Jang, Kuewhan; Lee, Sangmyung; Bang, Doyeon; Haam, Seungjoo; Choi, Chang-Hwan; Park, Jinsung; Na, Sungsoo

    2015-06-15

    Zinc oxide nanowires (ZnO NWs) have been attempted to various applications, such as piezoelectric devices, energy harvesting devices, self-powered nanosensors, and biomedical devices. However, recent reports have shown the toxic effect of ZnO NWs. In this report, we described the detection of ZnO NWs, for the first time using reduced graphene oxide (RGO) wrapping method. By wrapping RGO to ZnO NW (RGO-ZnO NW), we are able to aggregate ZnO NWs and increase the sensing performance. The detection measurement is based on the resonance frequency shift derived from mass variation of RGO-ZnO NW adsorption on the DNA immobilized resonator. The resonator is able to detect ZnO NWs with detection limit of 100 ng mL(-1) which is 2 order below the fatal toxic concentration of ZnO NWs in Human Monocyte Macrophages (HMMs). Furthermore, the resonator is able to detect ZnO NWs in real tap water, showing the potential as ZnO NWs screening platform in real environmental aqua system.

  7. Experimental investigation of rates and mechanisms of isotope exchange (O, H) between volcanic ash and isotopically-labeled water

    NASA Astrophysics Data System (ADS)

    Nolan, Gary S.; Bindeman, Ilya N.

    2013-06-01

    The hydrogen and oxygen isotope ratios in hydrous minerals and volcanic glass are routinely used as paleo-proxies to infer the isotopic values of meteoric waters and thus paleo-climatic conditions. We report a series of long-term exposure experiments of distal 7700 BP Mt. Mazama ash (-149‰ δ2H, +7‰ δ18O, 3.8 wt.% H2O) with isotopically-labeled water (+650‰ δ2H, +56‰ δ18O). Experiments were done at 70, 40 and 20 °C, and ranged in duration from 1 to 14454 h (˜20 months), to evaluate the rates of deuterium and 18O exchange, and investigate the relative role of exchange and diffusion. We also investigate the effect of drying on H2Otot and δ2H in native and reacted ash that can be used in defining the protocols for natural sample preparation. We employ Thermal Conversion Elemental Analyzer (TCEA) mass spectrometry, thermogravimetric analysis and a KBr pellet technique with infrared spectroscopy to measure the evolution of δ2H, total water, and OH water peaks in the course of exposure experiments, and in varying lengths of vacuum drying. Time series experiments aided by infrared measurements demonstrate the following new results: (i) It wasobserved that from 5 to >100‰ δ2H increases with time, with faster deuterium exchange at higher temperatures. Times at 15% of theoretical "full δ2H exchange" are: 15.8 years at 20 °C, 5.2 years at 40 °C, and 0.4 years at 70 °C. (ii) Even at extended exposure durations experiments show no net increase in water weight percent nor in δ18O in ash; water released from ash rapidly by thermal decomposition is not enriched in δ18O. This observation clearly suggests that it is hydrogen exchange, and not water addition or oxygen exchange that characterizes the process. (iii) Our time series drying, Fourier transform infrared (FTIR)-KBr and Thermogravimetric Analyzer (TGA) analyses collectively suggest a simple mechanistic view that there are three kinds of "water" in ash: water (mostly H2O) that is less strongly bonded

  8. Deuterium labeling of soil water movement in the Cuvelai-Etosha Basin, Namibia

    NASA Astrophysics Data System (ADS)

    Beyer, Matthias; Gaj, Marcel; Koeniger, Paul; Hamutoko, Josefina; Uugulu, Shoopi; Wanke, Heike; Lohe, Christoph; Himmelsbach, Thomas; Billib, Max

    2014-05-01

    Groundwater recharge estimations in semi-arid areas are challenging, especially in developing countries such as large parts of Sub-Saharan Africa, where data is generally scarce. Due to high heterogeneity in soil characteristics, vegetation and land use as well as spatially and temporally highly variable rainfall, precise site studies are necessary in order to characterize processes and quantify groundwater recharge rates. The stable isotope deuterium, 2H has been shown to be particularly suitable for such investigations. In this study, a field experiment using deuterium as an artificial tracer (2H2O, 70% deuterated water) was conducted to characterize movement of water during and after a synthetic rain event. The study was carried out in the framework of the project SASSCAL (Southern African Science Service Centre for Climate Change and Adaptive Land Management) in the Niipele catchment of the Cuvelai-Etosha Basin in Namibia at two locations differing in both soil and vegetation type: A forest site dominated by terminalia sericea, baikiaea plurijuga, burkea africana and acacia erioloba with deep pure sand soil and a shrub-/woodland site characterized by smaller burkea africana, borchemia discolor and acacia erioloba on a dark loamy sand soil underlain by a thick layer of calcrete. At both locations, soils were first saturated to trigger typical rainy season conditions and avoid immediate evaporation of the deuterated water. Subsequently, 500 ml of 2H2O was applied homogenously over a 0.25 m2 test plot at 25 cm depth. Finally, a 10 mm artificial rain event was applied onto the plot. Soil samples were collected every 10 cm to a maximum depth of 2.5 m with an Eijkelkamp hand auger after 1, 2, and 5 (respective 10) days. From these, soil water is extracted in the laboratory and subsequently analyzed for deuterium concentrations using a Picarro L2120-i cavity-ringdown (CRD) water vapor analyzer after vaporization. Additionally, grain size distribution, water content

  9. Shake-flask test for determination of biodegradation rates of (14)C-labeled chemicals at low concentrations in surface water systems.

    PubMed

    Ingerslev, F; Nyholm, N

    2000-03-01

    A simple shake-flask surface water biodegradability die away test with (14)C-labeled chemicals added to microgram per liter concentrations (usually 1-100 microg/L) is described and evaluated. The aim was to provide information on biodegradation behavior and kinetic rates at environmental (low) concentrations in surface water systems. The basic principle of measurement was to determine evolved CO(2) indirectly from measurements of total organic activity in subsamples after stripping off their content of CO(2). Used with surface water alone the test simulates a pelagic environment and amended with sediments (0.1-1 dry weight/L) the test is intended to simulate a water environment with suspended solids (e.g., resuspended sediments). A protocol of the test used with the (14)C technique or with specific chemical analysis was recently developed by the International Organization for Standardization. Practical experience with the method is presented for a set of reference substances. These substances could be ranked in five groups of decreasing biodegradability: aniline>p-nitrophenol, 2, 4-dichlorophenoxyacetic acid>4-chloroaniline>maleic hydrazide, pentachlorophenol>atrazine. It was found that degradation rates and lag periods varied considerably among sampling sites and sometimes also among samples from the same site. No significant correlation could be established between degradation rates and microbial biomass estimates. Even small portions of added sediments greatly enhanced biodegradation of the absorbable compound pentachlorophenol, probably by providing sites for microbial attachment. Repeated tests indicated consistent degradation behavior for the readily degradable substances, whereas degradation sometimes stopped or failed with the more recalcitrant substances. A preadaptation step involving regular reinoculation with freshly collected surface water could, however, overcome the problems of false-negative results.

  10. Water-induced degradation of polymer solar cells studied by H2(18)O labeling.

    PubMed

    Norrman, Kion; Gevorgyan, Suren A; Krebs, Frederik C

    2009-01-01

    Water-induced degradation of polymer photovoltaics based on the active materials poly(3-hexylthiophene) (P3HT) or poly[2-methoxy-5-(2'-ethylhexyloxy)-1,4-phenylenevinylene] (MEHPPV) was studied. The solar cell devices comprised a bulk heterojunction formed by the active material and [6,6]-phenyl-C61-butyric acid methyl ester (PCBM) in a standard device geometry. The use of H2(18)O in conjunction with time-of-flight secondary ion mass spectrometry enabled mapping of the parts of the device that were induced by water. A comparison was made between the two active materials and between devices that were kept in the dark and devices that had been subjected to illumination under simulated sunlight. Devices that were exposed to ambient humidity were compared to devices exposed to saturated humidity. Finally, a comparison was made between results obtained using H2(18)O and earlier work involving 18O2. Water was found to have behavior similar to but not identical with molecular oxygen.

  11. A New Method for Preparing Mesenchymal Stem Cells and Labeling with Ferumoxytol for Cell Tracking by MRI.

    PubMed

    Liu, Li; Tseng, Lanya; Ye, Qing; Wu, Yijen L; Bain, Daniel J; Ho, Chien

    2016-05-18

    Mesenchymal stem cells (MSCs) are among the major stem cells used for cell therapy and regenerative medicine. In-vivo cell-tracking by magnetic resonance imaging (MRI) is crucial for regenerative medicine, allowing verification that the transplanted cells reach the targeted sites. Cellular MRI combined with superparamagnetic iron-oxide (SPIO) contrast agents is an effective cell-tracking method. Here, we are reporting a new "bio-mimicry" method by making use of the "in-vivo environment" of MSCs to prepare native MSCs, so that (i) the phagocytic activity of cultured MSCs can be recovered and expanded MSCs can be ex-vivo labeled with Ferumoxytol, which is currently the only FDA approved SPIO nanoparticles for human use. Using our new method, 7-day cultured MSCs regain the capability to take up Ferumoxytol and exhibit an intracellular iron concentration of 2.50 ± 0.50 pg/MSC, comparable to that obtained by using Ferumoxytol-heparin-protamine nanocomplex; and (ii) cells can be re-sized to more native size, reducing from 32.0 ± 7.2 μm to 19.5 ± 5.2 μm. Our method can be very useful for expanding MSCs and labeling with Ferumoxytol, without the need for transfection agents and/or electroporation, allowing cell-tracking by MRI in both pre-clinical and clinical studies.

  12. A New Method for Preparing Mesenchymal Stem Cells and Labeling with Ferumoxytol for Cell Tracking by MRI

    PubMed Central

    Liu, Li; Tseng, Lanya; Ye, Qing; Wu, Yijen L.; Bain, Daniel J.; Ho, Chien

    2016-01-01

    Mesenchymal stem cells (MSCs) are among the major stem cells used for cell therapy and regenerative medicine. In-vivo cell-tracking by magnetic resonance imaging (MRI) is crucial for regenerative medicine, allowing verification that the transplanted cells reach the targeted sites. Cellular MRI combined with superparamagnetic iron-oxide (SPIO) contrast agents is an effective cell-tracking method. Here, we are reporting a new “bio-mimicry” method by making use of the “in-vivo environment” of MSCs to prepare native MSCs, so that (i) the phagocytic activity of cultured MSCs can be recovered and expanded MSCs can be ex-vivo labeled with Ferumoxytol, which is currently the only FDA approved SPIO nanoparticles for human use. Using our new method, 7-day cultured MSCs regain the capability to take up Ferumoxytol and exhibit an intracellular iron concentration of 2.50 ± 0.50 pg/MSC, comparable to that obtained by using Ferumoxytol-heparin-protamine nanocomplex; and (ii) cells can be re-sized to more native size, reducing from 32.0 ± 7.2 μm to 19.5 ± 5.2 μm. Our method can be very useful for expanding MSCs and labeling with Ferumoxytol, without the need for transfection agents and/or electroporation, allowing cell-tracking by MRI in both pre-clinical and clinical studies. PMID:27188664

  13. Evidence of negative energy balance using doubly labelled water in elite Kenyan endurance runners prior to competition.

    PubMed

    Fudge, Barry W; Westerterp, Klaas R; Kiplamai, Festus K; Onywera, Vincent O; Boit, Michael K; Kayser, Bengt; Pitsiladis, Yannis P

    2006-01-01

    Previous studies have found Kenyan endurance runners to be in negative energy balance during training and prior to competition. The aim of the present study was to assess energy balance in nine elite Kenyan endurance runners during heavy training. Energy intake and expenditure were determined over 7 d using weighed dietary intake and doubly labelled water, respectively. Athletes were on average in negative energy balance (mean energy intake 13 241 (SD 1330) kJ/d v. mean energy expenditure 14 611 (SD 1043) kJ/d; P=0.046), although there was no loss in body mass (mean 56.0 (SD 3.4) kg v. 55.7 (SD 3.6) kg; P=0.285). The calculation of underreporting was 13 % (range -24 to +9 %) and almost entirely accounted for by undereating (9 % (range -55 to +39 %)) as opposed to a lack of significant underrecording (i.e. total water intake was no different from water loss (mean 4.2 (SD 0.6) l/d v. 4.5 (SD 0.8) l/d; P=0.496)). Fluid intake was modest and consisted mainly of water (0.9 (SD 0.5) l/d) and milky tea (0.9 (SD 0.3) l/d). The diet was high in carbohydrate (67.3 (SD 7.8) %) and sufficient in protein (15.3 (SD 4.0) %) and fat (17.4 (SD 3.9) %). These results confirm previous observations that Kenyan runners are in negative energy balance during periods of intense training. A negative energy balance would result in a reduction in body mass, which, when combined with a high carbohydrate diet, would have the potential in the short term to enhance endurance running performance by reducing the energy cost of running.

  14. Comparison of electrical conductivity calculation methods for natural waters

    USGS Publications Warehouse

    McCleskey, R. Blaine; Nordstrom, D. Kirk; Ryan, Joseph N.

    2012-01-01

    The capability of eleven methods to calculate the electrical conductivity of a wide range of natural waters from their chemical composition was investigated. A brief summary of each method is presented including equations to calculate the conductivities of individual ions, the ions incorporated, and the method's limitations. The ability of each method to reliably predict the conductivity depends on the ions included, effective accounting of ion pairing, and the accuracy of the equation used to estimate the ionic conductivities. The performances of the methods were evaluated by calculating the conductivity of 33 environmentally important electrolyte solutions, 41 U.S. Geological Survey standard reference water samples, and 1593 natural water samples. The natural waters tested include acid mine waters, geothermal waters, seawater, dilute mountain waters, and river water impacted by municipal waste water. The three most recent conductivity methods predict the conductivity of natural waters better than other methods. Two of the recent methods can be used to reliably calculate the conductivity for samples with pH values greater than about 3 and temperatures between 0 and 40°C. One method is applicable to a variety of natural water types with a range of pH from 1 to 10, temperature from 0 to 95°C, and ionic strength up to 1 m.

  15. Fate of the (14)C-labeled herbicide prosulfocarb in a soil and in a sediment-water system.

    PubMed

    Braun, Karsten E; Luks, Ann-Katrin; Schmidt, Burkhard

    2017-02-01

    The fate of (14)C-labeled herbicide prosulfocarb was studied in an agricultural soil and in a sediment-water system, the sediment part of which was derived from Yangtze Three Gorges Reservoir, China. Time-course studies were performed for 28 d and 49 d, respectively. Main transformation routes of (14)C-prosulfocarb were mineralization to (14)CO2 and formation of nonextractable residues amounting to 12.13% and 10.43%, respectively, after 28 days (soil), and 9.40% and 11.98%, respectively, after 49 d (sediment-water system). Traces of prosulfocarbsulfoxide were detected by means of TLC, HPLC, and LC-MS; other transformation products were not found. Initial extraction of soil assays using 0.01 M CaCl2 solution showed that the bioavailability of the herbicide was considerably low; immediately after application (0.1 d of incubation), only 4.78% of applied radioactivity were detected in this aqueous fraction. DT50 values of (14)C-prosulfocarb estimated from radio-TLC and -HPLC analyses were above 28 d in soil and ranged between 29 d and 49 d in the sediment-water system. Partitioning of (14)C from water to sediment phase occurred with DT50 slightly above 2 d. With regard to the sediment-water system, adsorption occurred with log Koc = 1.38 (calculated from 2 day assays) and 2.35 (49 d assays). As similarly estimated from portions of (14)C found in CaCl2 extracts of the 0.1 d assays, (14)C-prosulfocarb's log Koc in soil was 2.96. With both experiments, similar portions of nonextractable radioactivity were associated with all soil organic matter fractions, i.e. nonhumics, fulvic acids, humic acids, and humin/minerals. Throughout all sample preparation, the experiments were severely impaired by losses of radioactivity especially with concentration of samples containing water in vacuo. All findings pointed to volatility of parent prosulfocarb in presence of water rather than volatility of transformation products. According to literature data, this behavior of

  16. Label-Free Fluorescence Assay of S1 Nuclease and Hydroxyl Radicals Based on Water-Soluble Conjugated Polymers and WS2 Nanosheets

    PubMed Central

    Li, Junting; Zhao, Qi; Tang, Yanli

    2016-01-01

    We developed a new method for detecting S1 nuclease and hydroxyl radicals based on the use of water-soluble conjugated poly[9,9-bis(6,6-(N,N,N-trimethylammonium)-fluorene)-2,7-ylenevinylene-co-alt-2,5-dicyano-1,4-phenylene)] (PFVCN) and tungsten disulfide (WS2) nanosheets. Cationic PFVCN is used as a signal reporter, and single-layer WS2 is used as a quencher with a negatively charged surface. The ssDNA forms complexes with PFVCN due to much stronger electrostatic interactions between cationic PFVCN and anionic ssDNA, whereas PFVCN emits yellow fluorescence. When ssDNA is hydrolyzed by S1 nuclease or hydroxyl radicals into small fragments, the interactions between the fragmented DNA and PFVCN become weaker, resulting in PFVCN being adsorbed on the surface of WS2 and the fluorescence being quenched through fluorescence resonance energy transfer. The new method based on PFVCN and WS2 can sense S1 nuclease with a low detection limit of 5 × 10−6 U/mL. Additionally, this method is cost-effective by using affordable WS2 as an energy acceptor without the need for dye-labeled ssDNA. Furthermore, the method provides a new platform for the nuclease assay and reactive oxygen species, and provides promising applications for drug screening. PMID:27304956

  17. Water turbine system and method of operation

    DOEpatents

    Costin, Daniel P [Montpelier, VT

    2011-05-10

    A system for providing electrical power from a current turbine is provided. The system includes a floatation device and a mooring. A water turbine structure is provided having an upper and lower portion wherein the lower portion includes a water fillable chamber. A plurality of cables are used to couple the system where a first cable couples the water turbine to the mooring and a second cable couples the floatation device to the first cable. The system is arranged to allow the turbine structure to be deployed and retrieved for service, repair, maintenance and redeployment.

  18. Water turbine system and method of operation

    DOEpatents

    Costin, Daniel P.

    2010-06-15

    A system for providing electrical power from a current turbine is provided. The system includes a floatation device and a mooring. A water turbine structure is provided having an upper and lower portion wherein the lower portion includes a water fillable chamber. A plurality of cables are used to couple the system where a first cable couples the water turbine to the mooring and a second cable couples the floatation device to the first cable. The system is arranged to allow the turbine structure to be deployed and retrieved for service, repair, maintenance and redeployment.

  19. Water turbine system and method of operation

    DOEpatents

    Costin, Daniel P [Montpelier, VT

    2009-02-10

    A system for providing electrical power from a current turbine is provided. The system includes a floatation device and a mooring. A water turbine structure is provided having an upper and lower portion wherein the lower portion includes a water fillable chamber. A plurality of cables are used to couple the system where a first cable couples the water turbine to the mooring and a second cable couples the floatation device to the first cable. The system is arranged to allow the turbine structure to be deployed and retrieved for service, repair, maintenance and redeployment.

  20. Food Labels

    MedlinePlus

    ... Loss Surgery? A Week of Healthy Breakfasts Shyness Food Labels KidsHealth > For Teens > Food Labels Print A ... have at least 95% organic ingredients. continue Making Food Labels Work for You The first step in ...

  1. Total Energy Expenditure in Obese Kuwaiti Primary School Children Assessed by the Doubly-Labeled Water Technique

    PubMed Central

    Davidsson, Lena; Al-Ghanim, Jameela; Al-Ati, Tareq; Al-Hamad, Nawal; Al-Mutairi, Anwar; Al-Olayan, Lulwa; Preston, Thomas

    2016-01-01

    The aim of this pilot study was to assess body composition and total energy expenditure (TEE) in 35 obese 7–9 years old Kuwaiti children (18 girls and 17 boys). Total body water (TBW) and TEE were assessed by doubly-labeled water technique. TBW was derived from the intercept of the elimination rate of deuterium and TEE from the difference in elimination rates of 18O and deuterium. TBW was used to estimate fat-free mass (FFM), using hydration factors for different ages and gender. Fat mass (FM) was calculated as the difference between body weight and FFM. Body weight was not statistically different but TBW was significantly higher (p = 0.018) in boys (44.9% ± 3.3%) than girls (42.4% ± 3.0%), while girls had significantly higher estimated FM (45.2 ± 3.9 weight % versus 41.6% ± 4.3%; p = 0.014). TEE was significantly higher in boys (2395 ± 349 kcal/day) compared with girls (1978 ± 169 kcal/day); p = 0.001. Estimated physical activity level (PAL) was significantly higher in boys; 1.61 ± 0.167 versus 1.51 ± 0.870; p = 0.034. Our results provide the first dataset of TEE in 7–9 years old obese Kuwaiti children and highlight important gender differences to be considered during the development of school based interventions targeted to combat childhood obesity. PMID:27754397

  2. Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

    PubMed

    Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

    2015-10-01

    White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.

  3. Method Development and Monitoring of Cyanotoxins in Water ...

    EPA Pesticide Factsheets

    Increasing occurrence of cyanobacterial harmful algal blooms (HABs) in ambient waters has become a worldwide concern. Numerous cyanotoxins can be produced during HAB events which are toxic to animals and humans. Validated standardized methods that are rugged, selective and sensitive are needed for these cyanotoxins in drinking and ambient waters. EPA Drinking Water Methods 544 (six microcystins [MCs] and nodularin) and 545 (cylindrospermopsin [CYL] and anatoxin-a [ANA]) have been developed using liquid chromatography/tandem mass spectrometry (LC/MS/MS). This presentation will describe the adaptation of Methods 544 and 545 to ambient waters and application of these ambient water methods to seven bodies of water across the country with visible cyanobacterial blooms.Several changes were made to Method 544 to accommodate the increased complexity of ambient water. The major changes were to reduce the sample volume from 500 to 100 mL for ambient water analyses and to incorporate seven additional MCs in an effort to capture data for more MC congeners in ambient waters. The major change to Method 545 for ambient water analyses was the addition of secondary ion transitions for each of the target analytes for confirmation purposes. Both methods have been ruggedly tested in bloom samples from multiple bodies of water, some with multiple sample locations and sampling days. For ambient water bloom samples spiked with MCs (>800 congener measurements), 97% of the measurements

  4. Simple method for preparation of fluor/hapten-labeled dUTP.

    PubMed

    Nimmakayalu, M; Henegariu, O; Ward, D C; Bray-Ward, P

    2000-03-01

    Many projects, such as multiplex-fluorescence in situ hybridization (M-FISH) karyotyping, require the use of relatively large amounts of multiple fluor- or hapten-labeled nucleotides for the preparation of DNA probes. Such a requirement makes these experimental approaches prohibitively expensive for many researchers. The cost of such nucleotides can be reduced approximately 99% by purchasing the chemical precursors, fluor or hapten succinimidyl esters and 5-(3-aminoallyl)-2'-deoxyuridine 5' triphosphate (AA-dUTP), and performing the simple coupling/purification described here. It is possible to finish four to ten different fluor/hapten dUTP preparations of 2.5 microM scale within a 24 h period. The reagent cost for each preparation ranges from $33-$237 per microM, depending on the fluor/hapten. This laboratory uses such nucleotide preparations to prepare FISH probes by nick translation or PCR amplification.

  5. Label-free relative quantification method for low-abundance glycoproteins in human serum by micrOTOF-Q.

    PubMed

    Hao, Piliang; Ren, Yan; Xie, Yongming

    2009-06-01

    In this study, a label-free relative quantification strategy was developed for quantifying low-abundance glycoproteins in human serum. It included three steps: (1) immunodepletion of 12 high-abundance proteins, (2) enrichment of low-abundance glycoproteins by multi-lectin column, (3) relative quantification of them between different samples by micrOTOF-Q. We also evaluated the specificity and efficiency of immunodepletion, the accuracy of protein quantification and the possible influence of immunodepletion, glycoprotein enrichment, trypsin digestion and peptide ionization on quantification. In conclusion, the relative quantification method can be effectively applied to the screening of low-abundance biomarkers.

  6. Two methods that facilitate autoradiography of small /sup 32/P-labeled DNA fragments following electrophoresis in agarose gels

    SciTech Connect

    Cockerill, P.N.

    1988-02-01

    Two methods which permit detection by autoradiography of small /sup 32/P-labeled DNA fragments resolved by agarose gel electrophoresis are described. Agarose gel electrophoresis poses problems for autoradiography as (i) the gels are normally too thick to allow autoradiography without being dried first, and (ii) fragments of DNA of 1000 bp or less in length are readily lost during drying. In this study DNA fragments as small as 121 bp have been retained in agarose gels upon drying. This has been achieved by either (i) first fixing the DNA with the cationic detergent cetyltrimethylammonium bromide, or (ii) drying the agarose gels onto Zeta-Probe charge-modified membranes.

  7. Human Health Water Quality Criteria and Methods for Toxics

    EPA Pesticide Factsheets

    Documents pertaining to Human Health Water Quality Criteria and Methods for Toxins. Includes 2015 Update for Water Quality Criteria, 2002 National Recommended Human Health Criteria, and 2000 EPA Methodology.

  8. EPA METHODS FOR VIRUS DETECTION IN WATER

    EPA Science Inventory

    A number of different types of human enteric viruses cause waterborne outbreaks when individuals are exposed to contaminated drinking and recreational waters. Members of the enterovirus group cause numerous diseases, including gastroenteritis, encephalitis, meningitis, myocard...

  9. EPA METHODS FOR VIRUS DETECTION IN WATER

    EPA Science Inventory

    A number of different types of human enteric viruses cause waterborne outbreaks when individuals are exposed to contaminated drinking and recreational waters. Members of the enterovirus group cause numerous diseases, including gastroenteritis, encephalitis, meningitis, myocard...

  10. Description of an Advantageous Optical Label-Free Biosensing Interferometric Read-Out Method to Measure Biological Species

    PubMed Central

    Holgado, Miguel; Sanza, Francisco J.; López, Ana; Lavín, Álvaro; Casquel, Rafael; Laguna, María F.

    2014-01-01

    In this article we report a new, simple, and reliable optical read-out detection method able to assess Rotavirus present in human sera as well as in the viral pollution sources. It is based on the interference of two interferometers used as biophotonic transducers. The method significantly improves the optical label-free biosensing response measuring both, the concentration of the AgR and its corresponding size. Two different immunoassays were carried out: Bovine Serum Albumin (BSA), and the recognition by its antibody (anti-BSA); and Rotavirus (AgR) and the recognition by its antibody (anti-AgR). In the cases studied, and using as model interferometer a simple Fabry-Perot transducer, we demonstrate a biosensing enhancement of two orders of magnitude in the Limit of Detection (LoD). In fact, this read-out optical method may have significant implications to enhance other optical label-free photonic transducers reported in the scientific literature. PMID:24566633

  11. Water quality assessment in Qu River based on fuzzy water pollution index method.

    PubMed

    Li, Ranran; Zou, Zhihong; An, Yan

    2016-12-01

    A fuzzy improved water pollution index was proposed based on fuzzy inference system and water pollution index. This method can not only give a comprehensive water quality rank, but also describe the water quality situation with a quantitative value, which is convenient for the water quality comparison between the same ranks. This proposed method is used to assess water quality of Qu River in Sichuan, China. Data used in the assessment were collected from four monitoring stations from 2006 to 2010. The assessment results show that Qu River water quality presents a downward trend and the overall water quality in 2010 is the worst. The spatial variation indicates that water quality of Nanbashequ section is the pessimal. For the sake of comparison, fuzzy comprehensive evaluation and grey relational method were also employed to assess water quality of Qu River. The comparisons of these three approaches' assessment results show that the proposed method is reliable. Copyright © 2016. Published by Elsevier B.V.

  12. Comparative analysis of statistical methods used for detecting differential expression in label-free mass spectrometry proteomics.

    PubMed

    Langley, Sarah R; Mayr, Manuel

    2015-11-03

    Label-free LC-MS/MS proteomics has proven itself to be a powerful method for evaluating protein identification and quantification from complex samples. For comparative proteomics, several methods have been used to detect the differential expression of proteins from such data. We have assessed seven methods used across the literature for detecting differential expression from spectral count quantification: Student's t-test, significance analysis of microarrays (SAM), normalised spectral abundance factor (NSAF), normalised spectral abundance factor-power law global error model (NSAF-PLGEM), spectral index (SpI), DESeq and QSpec. We used 2000 simulated datasets as well as publicly available data from a proteomic standards study to assess the ability of these methods to detect differential expression in varying effect sizes and proportions of differentially expressed proteins. At two false discovery rate (FDR) levels, we find that several of the methods detect differential expression within the data with reasonable precision, others detect differential expression at the expense of low precision, and finally, others which fail to identify any differentially expressed proteins. The inability of these seven methods to fully capture the differential landscape, even at the largest effect size, illustrates some of the limitations of the existing technologies and the statistical methodologies. In label-free mass spectrometry experiments, protein identification and quantification have always been important, but there is now a growing focus on comparative proteomics. Detecting differential expression in protein levels can inform on important biological mechanisms and provide direction for further study. Given the high cost and labour intensive nature of validation experiments, statistical methods are important for prioritising proteins of interest. Here, we have performed a comparative analysis to investigate the statistical methodologies for detecting differential expression

  13. Non-labeled monitoring of targeted liposome interactions with a model receptor surface: effect of flow rate and water content.

    PubMed

    Liang, Huamin; Tuppurainen, Jussi-Pekka; Lehtinen, Julia; Viitala, Tapani; Yliperttula, Marjo

    2013-11-20

    In this study, we present a novel in vitro approach that utilizes two surface-sensitive and label-free techniques, i.e. surface plasmon resonance (SPR) and quartz crystal microbalance (QCM), to study the interfacial events during liposome-target surface interactions. The flow channels of SPR and QCM devices were first synchronized via hydrodynamic modeling. Biotin-streptavidin was used as a model pair and self-assembled monolayers (SAMs) were utilized as model surfaces for targeted liposome-surface interaction studies. The interactions between biotin-liposomes and the streptavidin-biotin-SAM surfaces were investigated under controlled shear flows using the synchronized SPR and QCM devices. The response of the liposome interaction was monitored as a function of the flow rate. The affinity and the amount of bound liposome indicated that the increased flow rate improved the binding of the targeted liposomes to the model membrane surfaces. The combined use of the synchronized SPR and QCM devices for nanoparticle interaction studies clearly demonstrates the effect of the flow rate (or the shear stress) on the liposome binding. Our results suggest that the binding of liposomes to the model membranes is flow rate and shear stress regulated. Thus, the flow rate (or the shear stress), which is usually neglected, should be taken into account during the development and optimization of targeted liposome formulations. In addition, the water content within the liposome layer (including the water inside the liposomes and the water between the liposomes) had a significant influence on the visco-elasticity and the binding kinetics to the SAM surfaces.

  14. Quantitative Detection Method of Hydroxyapatite Nanoparticles Based on Eu(3+) Fluorescent Labeling in Vitro and in Vivo.

    PubMed

    Xie, Yunfei; Perera, Thalagalage Shalika Harshani; Li, Fang; Han, Yingchao; Yin, Meizhen

    2015-11-04

    One major challenge for application of hydroxyapatite nanoparticles (nHAP) in nanomedicine is the quantitative detection method. Herein, we exploited one quantitative detection method for nHAP based on the Eu(3+) fluorescent labeling via a simple chemical coprecipitation method. The trace amount of nHAP in cells and tissues can be quantitatively detected on the basis of the fluorescent quantitative determination of Eu(3+) ions in nHAP crystal lattice. The lowest concentration of Eu(3+) ions that can be quantitatively detected is 0.5 nM using DELFIA enhancement solution. This methodology can be broadly applicable for studying the tissue distribution and metabolization of nHAP in vivo.

  15. Rapid direct methods for enumeration of specific, active bacteria in water and biofilms

    NASA Technical Reports Server (NTRS)

    McFeters, G. A.; Pyle, B. H.; Lisle, J. T.; Broadaway, S. C.

    1999-01-01

    Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between

  16. Rapid direct methods for enumeration of specific, active bacteria in water and biofilms

    NASA Technical Reports Server (NTRS)

    McFeters, G. A.; Pyle, B. H.; Lisle, J. T.; Broadaway, S. C.

    1999-01-01

    Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between

  17. New effects in Langmuir and Langmuir-Blodgett monolayers from fluorescently labelled phospholipids and their possible use for water quality control

    NASA Astrophysics Data System (ADS)

    Ivanov, G. R.; Geshev, N. I.

    2016-02-01

    Secondary water contamination poses significant challenges to the sensitivity and selectivity of sensors used for its detection and monitoring. Currently only lab tests can detect these contaminants and by the time this happens the contaminated water has entered the city water supply system. Fluorescent chromophore NitroBenzoxaDiazole (NBD) is very suitable and had been successfully used in biosensor applications due to its high sensitivity to close proximity polarity of the medium. Over the years we have discovered 3 new effects in NBD- labelled phospholipids which can significantly improve the performance of biosensors. The phospholipid matrix provides flexible biocompatible environment for immobilization of selectively reacting enzymes, microorganisms, DNA, immunoagents, whole cells. Use of single layer (3.1 nm thickness) films at the air-water interface (Langmuir films) or deposited on solid support as Langmuir-Blodgett (LB) film gives fast response times for real time monitoring (no slow diffusion processes) and precise molecule ordering and orientation. The first new effect was fluorescence self-quenching in Langmuir and LB films. In the liquid phase films exhibit normal fluorescence. Upon transition to solid phase fluorescence intensity is almost completely self-quenched and fluorescence lifetimes in the nanosecond region decrease 2 times. This is easily measured. Usually large heavy metal atoms quench fluorescence. We observed the opposite new effect when LB film is deposited in the solid phase from a subphase containing heavy metals. The third new effect is the obtaining of nanosized cylinders with bilayer thickness, which remain stable at least for months, when LB monolayer is deposited in the phase coexistence region at thermodynamic equilibrium. This greatly increases reacting surface and sensitivity of possible sensors. Almost all possible optical experimental methods were used for this research. This includes polarized ATR FTIR and polarized UV

  18. Synchrotron X-ray footprinting as a method to visualize water in proteins

    SciTech Connect

    Gupta, Sayan; Feng, Jun; Chan, Leanne Jade G.; Petzold, Christopher J.; Ralston, Corie Y.

    2016-07-27

    The vast majority of biomolecular processes are controlled or facilitated by water interactions. In enzymes, regulatory proteins, membrane-bound receptors and ion-channels, water bound to functionally important residues creates hydrogen-bonding networks that underlie the mechanism of action of the macromolecule. High-resolution X-ray structures are often difficult to obtain with many of these classes of proteins because sample conditions, such as the necessity of detergents, often impede crystallization. Other biophysical techniques such as neutron scattering, nuclear magnetic resonance and Fourier transform infrared spectroscopy are useful for studying internal water, though each has its own advantages and drawbacks, and often a hybrid approach is required to address important biological problems associated with protein–water interactions. One major area requiring more investigation is the study of bound water molecules which reside in cavities and channels and which are often involved in both the structural and functional aspects of receptor, transporter and ion channel proteins. Recently, significant progress has been made in synchrotron-based radiolytic labeling and mass spectroscopy techniques for both the identification of bound waters and for characterizing the role of water in protein conformational changes at a high degree of spatial and temporal resolution. Finally, here the latest developments and future capabilities of this method for investigating water–protein interactions and its synergy with other synchrotron-based methods are discussed.

  19. Synchrotron X-ray footprinting as a method to visualize water in proteins

    DOE PAGES

    Gupta, Sayan; Feng, Jun; Chan, Leanne Jade G.; ...

    2016-07-27

    The vast majority of biomolecular processes are controlled or facilitated by water interactions. In enzymes, regulatory proteins, membrane-bound receptors and ion-channels, water bound to functionally important residues creates hydrogen-bonding networks that underlie the mechanism of action of the macromolecule. High-resolution X-ray structures are often difficult to obtain with many of these classes of proteins because sample conditions, such as the necessity of detergents, often impede crystallization. Other biophysical techniques such as neutron scattering, nuclear magnetic resonance and Fourier transform infrared spectroscopy are useful for studying internal water, though each has its own advantages and drawbacks, and often a hybrid approachmore » is required to address important biological problems associated with protein–water interactions. One major area requiring more investigation is the study of bound water molecules which reside in cavities and channels and which are often involved in both the structural and functional aspects of receptor, transporter and ion channel proteins. Recently, significant progress has been made in synchrotron-based radiolytic labeling and mass spectroscopy techniques for both the identification of bound waters and for characterizing the role of water in protein conformational changes at a high degree of spatial and temporal resolution. Finally, here the latest developments and future capabilities of this method for investigating water–protein interactions and its synergy with other synchrotron-based methods are discussed.« less

  20. Synchrotron X-ray footprinting as a method to visualize water in proteins

    PubMed Central

    Gupta, Sayan; Feng, Jun; Chan, Leanne Jade G.; Petzold, Christopher J.; Ralston, Corie Y.

    2016-01-01

    The vast majority of biomolecular processes are controlled or facilitated by water interactions. In enzymes, regulatory proteins, membrane-bound receptors and ion-channels, water bound to functionally important residues creates hydrogen-bonding networks that underlie the mechanism of action of the macromolecule. High-resolution X-ray structures are often difficult to obtain with many of these classes of proteins because sample conditions, such as the necessity of detergents, often impede crystallization. Other biophysical techniques such as neutron scattering, nuclear magnetic resonance and Fourier transform infrared spectroscopy are useful for studying internal water, though each has its own advantages and drawbacks, and often a hybrid approach is required to address important biological problems associated with protein–water interactions. One major area requiring more investigation is the study of bound water molecules which reside in cavities and channels and which are often involved in both the structural and functional aspects of receptor, transporter and ion channel proteins. In recent years, significant progress has been made in synchrotron-based radiolytic labeling and mass spectroscopy techniques for both the identification of bound waters and for characterizing the role of water in protein conformational changes at a high degree of spatial and temporal resolution. Here the latest developments and future capabilities of this method for investigating water–protein interactions and its synergy with other synchrotron-based methods are discussed. PMID:27577756

  1. Thermochemical method for producing hydrogen from water

    SciTech Connect

    Fujii, K.; Kondo, W.; Kumagai, T.

    1980-02-12

    A closed system for obtaining hydrogen from water is provided by combining a first step of obtaining hydrogen by reacting water and ferrous halide, a second step of converting triiron tetraoxide produced as a by-product in the first step to ferrous sulfate, a third step of obtaining oxygen and by-products by thermally decomposing said ferrous sulfate, and a fourth step of returning said by-products by thermally decomposing said ferrous sulfate, and a fourth step of returning said by-products obtained in the third step to any of the previous steps.

  2. Using doubly-labelled water to measure free-living energy expenditure: Some old things to remember and some new things to consider.

    PubMed

    Speakman, John R; Hambly, Catherine

    2016-12-01

    The doubly-labelled water (DLW) method provides the ability to measure the energy expenditure of free-living animals based only on the injection of two isotopes in water (one of oxygen and one of hydrogen) and traditionally the collection of 2 blood samples. We review here the fundamental basis of how the method works, and highlight how the choice of the appropriate calculation equation can have a large impact on the resultant estimates, particularly in species where the difference between the isotope elimination constants is small. This knowledge is not new, but is worth reiterating given the potential for error by making the wrong choice. In particular, it is important to remember that for mammals weighing less than 5kg, and birds weighing less than 2kg, the single pool models perform best in validation studies, while in mammals above 15kg the two-pool models perform best. Above 2kg in birds and between 5 and 15kg in mammals, however, the model superiority is uncertain. Even where the choice based on body mass would appear clear, the decision may need to be tempered by species specific information regarding potential additional sources for hydrogen turnover, such as de novo lipogenesis or methanogenesis. Recent advances in the technique have included attempts to make the method less invasive by using innovative methods for dosing and sample collection. In addition, the advent of laser spectroscopy, as a replacement technology for mass spectrometry, may open up many new opportunities in the field. These potentially include direct sampling of breath in the field and tracking background isotope drift using (17)oxygen levels. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. A facile label-free G-quadruplex based fluorescent aptasensor method for rapid detection of ATP

    NASA Astrophysics Data System (ADS)

    Liu, Haisheng; Ma, Changbei; Ning, Feng; Chen, Hanchun; He, Hailun; Wang, Kemin; Wang, Jun

    2017-03-01

    The present work demonstrates a simple, rapid and label-free ATP detection method using a fluorescent aptasensor that is based on G-quadruplex formation. In the absence of ATP, the Thioflavin T (ThT) dye binds to the G-rich ATP aptamer and forms an ATP aptamer/ThT G-quadruplex complex, which results in high fluorescence intensity. Upon addition of ATP, the ATP aptamer/ThT complex will be replaced by the formation of an ATP aptamer/ATP complex. During this process, separation of the ThT dye from the ATP aptamer/ThT complex decreases the fluorescence intensity of the reaction mixture dramatically. This fluorescence aptasensor is highly sensitive and rapid, with a detection limit of 18 nM and a total reaction time of only 10 min. Furthermore, this method is cost-effective and simple, removing the requirement for labeling the detection reagents with a fluorophore-quencher pair.

  4. Comparing the Ability of Enhanced Sampling Molecular Dynamics Methods To Reproduce the Behavior of Fluorescent Labels on Proteins.

    PubMed

    Walczewska-Szewc, Katarzyna; Deplazes, Evelyne; Corry, Ben

    2015-07-14

    Adequately sampling the large number of conformations accessible to proteins and other macromolecules is one of the central challenges in molecular dynamics (MD) simulations; this activity can be difficult, even for relatively simple systems. An example where this problem arises is in the simulation of dye-labeled proteins, which are now being widely used in the design and interpretation of Förster resonance energy transfer (FRET) experiments. In this study, MD simulations are used to characterize the motion of two commonly used FRET dyes attached to an immobilized chain of polyproline. Even in this simple system, the dyes exhibit complex behavior that is a mixture of fast and slow motions. Consequently, very long MD simulations are required to sufficiently sample the entire range of dye motion. Here, we compare the ability of enhanced sampling methods to reproduce the behavior of fluorescent labels on proteins. In particular, we compared Accelerated Molecular Dynamics (AMD), metadynamics, Replica Exchange Molecular Dynamics (REMD), and High Temperature Molecular Dynamics (HTMD) to equilibrium MD simulations. We find that, in our system, all of these methods improve the sampling of the dye motion, but the most significant improvement is achieved using REMD.

  5. Apparatus and method for reading two-dimensional electrophoretograms containing. beta. -ray-emitting labeled compounds

    SciTech Connect

    Anderson, H.L.; Kinnison, W.W.; Lillberg, J.W.

    1987-08-11

    An apparatus for reading flat, substantially planar two-dimensional gel electrophoretograms containing ..beta..-ray-emitting labeled compounds, the apparatus comprising in combination: a. a single, substantially flat and rectangular proportional chamber radiation detection means substantially parallel to and approximately coextensive with the gel electrophoretogram for receiving ..beta..-rays emitted from the electrophoretogram and cosmic rays and generating electrical signals according to the two dimensional coordinates of each ..beta..-ray and cosmic ray received thereby; b. substantially flat and rectangular scintillator means located nearby to the substantially flat and rectangular proportional chamber radiation detection means for detecting the incidence of cosmic rays on the substantially flat and rectangular proportional chamber radiation detection means and having an electrical output in response thereto, yet having substantially no sensitivity to the ..beta..-rays emitted from the gel; c. magnetic field generating means for providing a substantially uniform magnetic field having dimensions substantially coextensive with the rectangular dimension of the substantially flat and rectangular proportional chamber radiation detection means and passing substantially perpendicularly therethrough and through the scintillator means.

  6. Label-free method for cell counting in crude biological samples via paramagnetic bead aggregation.

    PubMed

    Li, Jingyi; Liu, Qian; Xiao, Li; Haverstick, Doris M; Dewald, Alison; Columbus, Linda; Kelly, Kimberly; Landers, James P

    2013-12-03

    Under chaotropic conditions, DNA released from lysed cells causes the aggregation of paramagnetic beads in a rotating magnetic field in a manner that is independent of the presence of other cellular components. The extent of aggregation correlates with the mass of DNA in a quantitative manner (Leslie, D. C. et al., J. Am. Chem. Soc. 2012, 134, 5689-96), and from this, the number of DNA-containing cells in the sample can be enumerated. Microbial growth testing is demonstrated by monitoring bead aggregation with E. coli in the presence of ampicillin. Without the need for fluorescent labeling or Coulter counting, the white blood cell count can be defined directly from a microliter of crude whole blood. Specificity is brought to the process by coupling bead-based immunocapture with DNA-bead aggregation allowing for the enumeration of CD4+ T cells from human blood samples. The results of DNA-induced bead aggregation had a 95% correlation with those generated by flow cytometry. With the process requiring only inexpensive, widely available benchtop laboratory hardware, a digital camera, and a simple algorithm, this provided a highly accessible alternative to more expensive cell-counting techniques.

  7. A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes

    PubMed Central

    Shahidi, Réza; Williams, Elizabeth A; Conzelmann, Markus; Asadulina, Albina; Verasztó, Csaba; Jasek, Sanja; Bezares-Calderón, Luis A; Jékely, Gáspár

    2015-01-01

    Electron microscopy-based connectomics aims to comprehensively map synaptic connections in neural tissue. However, current approaches are limited in their capacity to directly assign molecular identities to neurons. Here, we use serial multiplex immunogold labeling (siGOLD) and serial-section transmission electron microscopy (ssTEM) to identify multiple peptidergic neurons in a connectome. The high immunogenicity of neuropeptides and their broad distribution along axons, allowed us to identify distinct neurons by immunolabeling small subsets of sections within larger series. We demonstrate the scalability of siGOLD by using 11 neuropeptide antibodies on a full-body larval ssTEM dataset of the annelid Platynereis. We also reconstruct a peptidergic circuitry comprising the sensory nuchal organs, found by siGOLD to express pigment-dispersing factor, a circadian neuropeptide. Our approach enables the direct overlaying of chemical neuromodulatory maps onto synaptic connectomic maps in the study of nervous systems. DOI: http://dx.doi.org/10.7554/eLife.11147.001 PMID:26670546

  8. Metabolic fate of the (14)C-labeled herbicide clodinafop-propargyl in a sediment-water system.

    PubMed

    Yuan, Ye; Weitzel, Pascal; Schäffer, Andreas; Schmidt, Burkhard

    2015-01-01

    The metabolic fate of (14)C-phenyl-labeled herbicide clodinafop-propargyl ((14)C-CfP) was studied for 28 days in lab assays using a sediment-water system derived from a German location. Mineralization was 5.21% of applied (14)C after 28 days exhibiting a distinct lag phase until day 14 of incubation. Portions of radioactivity remaining in water phases decreased at moderate rate to 18.48% after 28 days; 62.46% were still detected in water after 14 days. Soxhlet extraction of the sediment using acetonitrile released 35.56% of applied (14)C with day 28, while 33.99% remained as non-extractable residues. A remarkable increase of bound (14)C was observed between 14 and 28 days correlating with the distinct increase of mineralization. No correlation was found throughout incubation with microbial activity of the sediment as determined by dimethyl sulfoxide reduction. Dissolved oxygen and pH value of water phases remained almost constant for 28 days. Analyses of Soxhlet extracts of the sediment and ethyl acetate extracts of water phases by radio-TLC and radio-HPLC revealed that CfP was rapidly cleaved to free acid clodinafop (Cf), which was further (bio-) transformed. DT50 values (based on radio-HPLC) were below 1 day (CfP) and slightly above 28 days (Cf). Further metabolites were not detected. Fractionation of humic and non-humic components of the sediment demonstrated that CfP's non-extractable residues were predominantly associated with fulvic acids up to 14 days of incubation (3.36%), whereas after 28 days, the majority of radioactivity was found in the humin/mineral fraction (13.30% of applied (14)C). Due to high-performance size-exclusion chromatography of the fulvic acids fraction derived from assays incubated for 28 days, this portion of (14)C was firmly, possibly covalently bound to fulvic acids and did not consist of CfP or Cf. Using an isolation strategy comprising preincubation of sediment with CfP and mineralization of (14)C-CfP as criterion, a microorganism

  9. A method for evaluating the host range of bacteriophages using phages fluorescently labeled with 5-ethynyl-2'-deoxyuridine (EdU).

    PubMed

    Ohno, Sayaka; Okano, Hironori; Tanji, Yasunori; Ohashi, Akiyoshi; Watanabe, Kazuya; Takai, Ken; Imachi, Hiroyuki

    2012-08-01

    The evaluation of bacteriophage (phage) host range is a significant issue in understanding phage and prokaryotic community interactions. However, in conventional methods, such as plaque assay, target host strains must be isolated, although almost all environmental prokaryotes are recalcitrant to cultivation. Here, we introduce a novel phage host range evaluation method using fluorescently labeled phages (the FLP method), which consists of the following four steps: (i) Fluorescently labeled phages are added to a microbial consortium, and host cells are infected and fluorescently labeled. (ii) Fluorescent cells are sorted by fluorescence-activated cell sorting. (iii) 16S rRNA gene sequences retrieved from sorted cells are analyzed, and specific oligonucleotide probes for fluorescence in situ hybridization (FISH) are designed. (iv) Cells labeled with both fluorescently labeled phage and FISH probe are identified as host cells. To verify the feasibility of this method, we used T4 phage and Escherichia coli as a model. We first used nucleic acid stain reagents for phage labeling; however, the reagents also stained non-host cells. Next, we employed the Click-iT EdU (5-ethynyl-2'-deoxyuridine) assay kit from Invitrogen for phage labeling. Using EdU-labeled T4 phage, we could specifically detect E. coli cells in a complex microbial consortium from municipal sewage. We also confirmed that FISH could be applied to the infected E. coli cells. These results suggest that this FLP method using the EdU assay kit is a useful method for evaluating phage host range and may have a potential application for various types of phages, even if their prokaryotic hosts are currently unculturable.

  10. Other Clean Water Act Test Methods: Biosolids

    EPA Pesticide Factsheets

    Methods for analysis of chemical pollutants in biosolids (municipal sewage sludge). These methods are not approved under 40 CFR Part 136, but may be of interest to regulated entities, permitting authorities and the public.

  11. Evaluation of Chemical and Microbiological Quality in 21 Brands of Iranian Bottled Drinking Waters in 2012: A Comparison Study on Label and Real Contents

    PubMed Central

    Moazeni, M.; Atefi, M.; Ebrahimi, A.; Razmjoo, P.; Vahid Dastjerdi, M.

    2013-01-01

    The purpose of this study was to evaluate and compare chemical and microbiological quality of the 21 Iranian bottled drinking waters reported on manufacturer's labeling and standards in 2012. Samples were analyzed for chemical properties K+, F−, SO4 2−, Cl−, Mg2+, Ca2+, and pH. Total and fecal coliform and heterotrophic plate counts of selected samples were analyzed by MPN and HPC tests, respectively, for three months. Finally, the labeled and real contents of the samples were compared. Potassium and sulfate ions about 43 and 52 percent of studied sample contents had values higher than label amounts, respectively. Ca2+, Cl− ions, and pH were about 71, 48, and 67 percent, respectively, less than label values. Total and fecal coliforms had negative results. The mean concentrations and standard deviations for K+, Cl−, pH, Ca2+, Mg2+, SO4 2−, and HPC were 1.13 ± 1.06, 16.39 ± 31.97, 6.6 ± 0.7, 28.35 ± 10.34, 86.58 ± 33.21, 24.17 ± 17.30 mg/L, and 16855 ± 25603 cfu/mL, respectively. Thus, there is possibility of microorganisms' growth in favorite conditions in bottled water. It was imperative to assess the public health risks in bottled water in Iran. PMID:23690802

  12. How to save water by choice of irrigation application method

    USDA-ARS?s Scientific Manuscript database

    It is known that irrigation application method can impact crop water use and water use efficiency, but the mechanisms involved are incompletely understood, particularly in terms of the water and energy balances during the growing season from pre-irrigation through planting, early growth and yield de...

  13. Evaluation of a high-intensity focused ultrasound-immobilized trypsin digestion and 18O-labeling method for quantitative proteomics.

    PubMed

    López-Ferrer, Daniel; Hixson, Kim K; Smallwood, Heather; Squier, Thomas C; Petritis, Konstantinos; Smith, Richard D

    2009-08-01

    A new method that uses immobilized trypsin concomitant with ultrasonic irradiation results in ultrarapid digestion and more thorough (18)O labeling for quantitative protein comparisons. The method was reproducible and provided effective digestions within <1 min with lower amounts of enzyme, compared to traditional methods. This method was demonstrated for digestion of both simple and complex protein mixtures, including bovine serum albumin, a global proteome extract from the bacteria Shewanella oneidensis, and mouse plasma, as well as (18)O labeling of complex protein mixtures, validating this method for differential proteomic measurements. This approach is simple, reproducible, cost-effective, rapid, and well suited for automation.

  14. Method for thermochemical decomposition of water

    DOEpatents

    Abraham, Bernard M.; Schreiner, Felix

    1977-01-11

    Water is thermochemically decomposed to produce hydrogen by the following sequence of reactions: KI, NH.sub.3, CO.sub. 2 and water in an organic solvent such as ethyl or propyl alcohol are reacted to produce KHCO 3 and NH.sub.4 I. The KHCO.sub.3 is thermally decomposed to K.sub.2 CO.sub.3, H.sub.2 O and CO.sub.2, while the NH.sub.4 I is reacted with Hg to produce HgI.sub.2, NH.sub.3 and H.sub.2. The K.sub.2 CO.sub.3 obtained by calcining KHCO.sub.3 is then reacted with HgI.sub.2 to produce Hg, KI, CO and O.sub.2. All products of the reaction are recycled except hydrogen and oxygen.

  15. A rapid improved method for gamma-spectrometric determination of 202Tl impurities in [201Tl]-labelled radiopharmaceuticals.

    PubMed

    Bonardi, Mauro; Groppi, Flavia; Birattari, Claudio

    2002-11-01

    Despite the cyclotron production method and the efficiency of the radiochemical procedures adopted, the long-lived radio-isotopic impurity 202Tl is always present in [201Tl]-labelled radio-pharmaceuticals, together with other short-lived impurities like, 200Tl. Rapid determination of the 202Tl impurity, can be achieved using HPGe gamma spectrometry and a detector shielded by a 5 mm thick envelope of lead. In this way, dead-time correction errors, Compton and X-ray background, are very efficiently avoided and suppressed. The same method could be applied routinely in nuclear medicine, to determine the radioisotopic purity of 201Tl by means of an ionisation chamber dose calibrator.

  16. A sensitive and selective resonance Rayleigh scattering method for quick detection of avidin using affinity labeling Au nanoparticles

    NASA Astrophysics Data System (ADS)

    Wang, Qi; Huang, Xi; Fu, Xuan; Deng, Huan; Ma, Meihu; Cai, Zhaoxia

    2016-06-01

    Avidin is a glycoprotein with antinutritional property, which should be limited in daily food. We developed an affinity biosensor system based on resonance Rayleigh scattering (RRS) and using affinity biotin labeling Au nanoparticles (AuNPs). This method was selective and sensitive for quick avidin detection due to the avidin-biotin affinitive interaction. Under optimal conditions, RRS intensity of biotin-AuNPs increase linearly with an increasing concentration of avidin from 5 to 160 ng/mL. The lower limit of detection was 0.59 ng/mL. This rapid and selective avidin detection method was used in synthetic samples and egg products with recoveries of between 102.97 and 107.92%, thereby demonstrating the feasible and practical application of this assay.

  17. Simple Method of Synthesizing Nickel-Nitrilotriacetic Acid Gold Nanoparticles with a Narrow Size Distribution for Protein Labeling

    NASA Astrophysics Data System (ADS)

    Kitai, Toshiyuki; Watanabe, Yuta; Toyoshima, Yoko Y.; Kobayashi, Takuya; Murayama, Takashi; Sakaue, Hiroyuki; Suzuki, Hitoshi; Takahagi, Takayuki

    2011-09-01

    We developed a simple method to synthesize nickel-nitrilotriacetic acid gold nanoparticles (Ni-NTA Au NPs) with a narrow size distribution for site-specific labeling in protein complexes. Au NPs were synthesized by the reduction of HAuCl4 using trisodium citrate and tannin acid. Then, the nanoparticle surfaces were modified with NTA and subsequent complexation with Ni2+. The mean diameter of the synthesized Ni-NTA Au NPs was 4.3 nm, and the coefficient of variation was 9%. The specific binding of the Ni-NTA Au NPs to polyhistidine-tagged (His-tagged) proteins was determined by transmission electron microscopy using kinesin and the p62 subunit of dynactin. Consequently, our method is useful for analyzing the substructures of protein complexes.

  18. A sensitive and selective resonance Rayleigh scattering method for quick detection of avidin using affinity labeling Au nanoparticles.

    PubMed

    Wang, Qi; Huang, Xi; Fu, Xuan; Deng, Huan; Ma, Meihu; Cai, Zhaoxia

    2016-06-05

    Avidin is a glycoprotein with antinutritional property, which should be limited in daily food. We developed an affinity biosensor system based on resonance Rayleigh scattering (RRS) and using affinity biotin labeling Au nanoparticles (AuNPs). This method was selective and sensitive for quick avidin detection due to the avidin-biotin affinitive interaction. Under optimal conditions, RRS intensity of biotin-AuNPs increase linearly with an increasing concentration of avidin from 5 to 160 ng/mL. The lower limit of detection was 0.59 ng/mL. This rapid and selective avidin detection method was used in synthetic samples and egg products with recoveries of between 102.97 and 107.92%, thereby demonstrating the feasible and practical application of this assay.

  19. Alternative cooling tower water treatment methods

    SciTech Connect

    Wilsey, C.A.

    1996-11-01

    The factors that contribute to proper water balance include total alkalinity, calcium hardness, and pH. In order to keep the cooling tower from scaling or corroding, a manipulation of these components is often necessary. This has traditionally been achieved with the use of chemicals, including but not limited to the following: acid, soda ash, sodium bicarbonate, calcium bicarbonate, algicide, and bactericide. Extensive research has shown that a balanced water system can also be achieved by using the proper combination of copper with a known halogen. Microbiologists have determined that a small amount of copper, acting as a supplement to chlorine at 0.4 ppm, has the same efficiency as 2.0 ppm free chlorine. Therefore, by using the following combination of components and procedures, the desired results can still be achieved: production of copper compound ions as a supplement to the chemical regimen; analysis and manipulation of make-up water; the use of copper as a coagulant for reduction of scale; copper as a supplemental bacterial disinfectant; and copper as an algicide.

  20. A fluorophore-labelled copper complex: crystal structure, hybrid cyclic water-perchlorate cluster and biological properties.

    PubMed

    Bhat, Satish S; Revankar, Vidyanand K; Shivalingegowda, Naveen; Lokanath, N K

    2017-09-01

    A fluorophore-labelled copper(II) complex, aquabis(dimethylformamide-κO)(perchlorato-κO)[2-(quinolin-2-yl)-1,3-oxazolo[4,5-f][1,10]phenanthroline]copper(II) perchlorate monohydrate, [Cu(ClO4)(C22H12N4O)(C3H7NO)2(H2O)]ClO4·H2O, has been synthesized and characterized. A cyclic hydrogen-bonded water-perchlorate anionic cluster, i.e. [(ClO4)2(H2O)2](2-), has been identified within the structure. Each cyclic anionic cluster unit is interconnected by hydrogen bonding to the cation. The cations join into an infinite hydrogen-bonded chain running in the [010] direction. Furthermore, interaction of the complex with calf-thymus DNA (CT-DNA) and cellular localization within the cells was explored. Spectroscopic studies indicate that the compound has a good affinity for DNA and stains the nucleus of the cells.

  1. Comparison of energy expenditure by the doubly labeled water technique with energy intake, heart rate, and activity recording in man

    SciTech Connect

    Schulz, S.; Westerterp, K.R.; Brueck, K.

    1989-06-01

    Average daily energy expenditure determined by the doubly labeled water technique (dlwEE) was compared in six subjects (aged 20-30 y) over 2 wk under usual living conditions; average food energy intake and energy expenditure estimated from individual diary records of physical activity. In addition, energy expenditure was estimated from 24-h heart rate recordings carried out on two randomly chosen days of the 2-wk period. The group means of the dlwEE were 1.94 +/- 0.24 (means +/- SD) times larger than resting metabolic rate (= 1.94 met) and nearly identical to the average daily energy intake (1.93 +/- 0.23 met). Energy expenditure estimated from the diaries of activity and from the 24-h heart rate recording varied between 1.67 and 2.24 met depending on the calculation procedure. The dlwEE (1.94 +/- 0.24 met) is much higher than that recently determined for sedentary people (1.25 met) and thus explains that young students may achieve body weight balance with a relatively high daily food energy intake.

  2. DNA hybridization detection with water-soluble conjugated polymers and chromophore-labeled single-stranded DNA.

    PubMed

    Gaylord, Brent S; Heeger, Alan J; Bazan, Guillermo C

    2003-01-29

    A sensor is provided that detects single-stranded deoxyribonucleic acid (ssDNA) with a specific base sequence. The ssDNA sequence sensor comprises an aqueous solution containing a cationic water-soluble conjugated polymer [in this case, poly(9,9-bis(6'-N,N,N-trimethylammonium)-hexyl)-fluorene phenylene), 1] with a ssDNA labeled with a dye (in this case, fluorescein). The emission of light from the sensor solution with the wavelength characteristic of the probe oligonucleotide indicates the presence of ssDNA with a specific base sequence complementary to that of the probe ssDNA-fluorescein. Maximum energy transfer from 1 to the signaling chromophore occurs when the ratio of polymer chains to DNA strands is approximately 1:1. Energy transfer from 1 results in a fluorescein emission that is more intense than that observed by direct excitation of the chromophore. Furthermore, the decrease in energy transfer upon addition of electrolyte indicates that electrostatic forces dominate the interactions between 1 and DNA.

  3. Measurement of myocardial blood flow with oxygen-15 labelled water: comparison of different administration protocols.

    PubMed

    Hermansen, F; Rosen, S D; Fath-Ordoubadi, F; Kooner, J S; Clark, J C; Camici, P G; Lammertsma, A A

    1998-07-01

    Positron emission tomography (PET) in conjunction with C15O2 or H215O can be used to measure myocardial blood flow (MBF) and tissue fraction (TF), i.e. the fraction of the tissue mass in the volume of the region of interest. However, with C15O2 inhalation, the tissue fraction in the septum is overestimated. Bolus injection of H215O together with arterial cannulation gives very precise results but is invasive. The purpose of this study was to develop a method which circumvents these problems. A four-parameter model with parameters for MBF, TF and spill-over fractions from both left and right ventricular cavities was developed. This method was compared with a three-parameter model (no right ventricular cavity spill-over) in both septal and non-septal regions of interest for three different administration protocols: bolus injection of H215O, infusion of H215O and inhalation of C15O2. It was found that MBF can be measured with intravenous administration of H215O without the requirement for arterial cannulation. The four-parameter protocol with bolus injection was stable in clinical studies. The four-parameter model proved essential for the septum, where it gave highly significantly better fits than did the three-parameter model (P<0.00003 in each of 15 subjects). Administration of H215O together with this four-parameter model also circumvented the problem of overestimation of TF in the septum seen with C15O2 inhalation. In addition, the radiation dose of H215O protocols is lower than that of C15O2 inhalation. Using a left atrial input curve instead of a left ventricular cavity input curve gave the same mean MBF and TF.

  4. Complex admixtures of clathrate hydrates in a water desalination method

    DOEpatents

    Simmons, Blake A.; Bradshaw, Robert W.; Dedrick, Daniel E.; Anderson, David W.

    2009-07-14

    Disclosed is a method that achieves water desalination by utilizing and optimizing clathrate hydrate phenomena. Clathrate hydrates are crystalline compounds of gas and water that desalinate water by excluding salt molecules during crystallization. Contacting a hydrate forming gaseous species with water will spontaneously form hydrates at specific temperatures and pressures through the extraction of water molecules from the bulk phase followed by crystallite nucleation. Subsequent dissociation of pure hydrates yields fresh water and, if operated correctly, allows the hydrate-forming gas to be efficiently recycled into the process stream.

  5. An innovative, quick and convenient labeling method for the investigation of pharmacological behavior and the metabolism of poly(DL-lactide-co-glycolide) nanospheres

    NASA Astrophysics Data System (ADS)

    Stevanović, Magdalena; Maksin, Tatjana; Petković, Jana; Filipič, Metka; Uskoković, Dragan

    2009-08-01

    Nanoparticles of poly(DL-lactide-co-glycolide) (PLGA) in the size range 90-150 nm were produced using the physicochemical method with solvent/non-solvent systems. The encapsulation of the ascorbic acid in the polymer matrix was performed by homogenization of the water and organic phases. In vitro degradation and release tests of PLGA nanoparticles with and without encapsulated ascorbic acid were studied for more than 60 days in PBS and it has been determined that PLGA completely degrades within this period, fully releasing all encapsulated ascorbic acid. The cytotoxicity of PLGA and PLGA/ascorbic acid 85/15% nanoparticles was examined with human hepatoma cell lines (HepG2 ECACC), in vitro. The obtained results indicate that neither PLGA nanospheres nor PLGA/ascorbic acid 85/15% nanoparticles significantly affected the viability of the HepG2 cells. The investigation of the distribution and pharmacokinetics of PLGA is crucial for the effective prediction of host responses to PLGA in particular applications. Thus we present a method of labeling PLGA nanospheres and PLGA/ascorbic acid 85/15 wt% nanoparticles by 99mTc which binds outside, leaving the cage intact. This enables a quick and convenient investigation of the pharmacological behavior and metabolism of PLGA. The biodistribution of 99mTc-labeled PLGA particles with and without encapsulated ascorbic acid after different periods of time of their installation into rats was examined. PLGA nanospheres with encapsulated ascorbic acid exhibit prolonged blood circulation accompanied by time-dependent reduction in the lungs, liver and spleen, and addition in the kidney, stomach and intestine. The samples were characterized by x-ray diffraction, scanning electron microscopy, stereological analysis, transmission electron microscopy, ultraviolet spectroscopy and instant thin layer chromatography.

  6. Methods for collection and analysis of water samples

    USGS Publications Warehouse

    Rainwater, Frank Hays; Thatcher, Leland Lincoln

    1960-01-01

    This manual contains methods used by the U.S. Geological Survey to collect, preserve, and analyze water samples. Throughout, the emphasis is on obtaining analytical results that accurately describe the chemical composition of the water in situ. Among the topics discussed are selection of sampling sites, frequency of sampling, field equipment, preservatives and fixatives, analytical techniques of water analysis, and instruments. Seventy-seven laboratory and field procedures are given for determining fifty-three water properties.

  7. Measuring Plant Water Status: A Simple Method for Investigative Laboratories.

    ERIC Educational Resources Information Center

    Mansfield, Donald H.; Anderson, Jay E.

    1980-01-01

    Describes a method suitable for quantitative studies of plant water status conducted by high school or college students and the calculation of the relative water content (RWC) of a plant. Materials, methods, procedures, and results are discussed, with sample data figures provided. (CS)

  8. Measuring Plant Water Status: A Simple Method for Investigative Laboratories.

    ERIC Educational Resources Information Center

    Mansfield, Donald H.; Anderson, Jay E.

    1980-01-01

    Describes a method suitable for quantitative studies of plant water status conducted by high school or college students and the calculation of the relative water content (RWC) of a plant. Materials, methods, procedures, and results are discussed, with sample data figures provided. (CS)

  9. The Leucine Incorporation Method Estimates Bacterial Growth Equally Well in Both Oxic and Anoxic Lake Waters

    PubMed Central

    Bastviken, David; Tranvik, Lars

    2001-01-01

    Bacterial biomass production is often estimated from incorporation of radioactively labeled leucine into protein, in both oxic and anoxic waters and sediments. However, the validity of the method in anoxic environments has so far not been tested. We compared the leucine incorporation of bacterial assemblages growing in oxic and anoxic waters from three lakes differing in nutrient and humic contents. The method was modified to avoid O2 contamination by performing the incubation in syringes. Isotope saturation levels in oxic and anoxic waters were determined, and leucine incorporation rates were compared to microscopically observed bacterial growth. Finally, we evaluated the effects of O2 contamination during incubation with leucine, as well as the potential effects of a headspace in the incubation vessel. Isotope saturation occurred at a leucine concentration of above about 50 nM in both oxic and anoxic waters from all three lakes. Leucine incorporation rates were linearly correlated to observed growth, and there was no significant difference between oxic and anoxic conditions. O2 contamination of anoxic water during 1-h incubations with leucine had no detectable impact on the incorporation rate, while a headspace in the incubation vessel caused leucine incorporation to increase in both anoxic and O2-contaminated samples. The results indicate that the leucine incorporation method relates equally to bacterial growth rates under oxic and anoxic conditions and that incubation should be performed without a headspace. PMID:11425702

  10. Labeling technique for nonplanar surfaces based on the combination of a diffractive axilens with digital holography methods

    NASA Astrophysics Data System (ADS)

    Marin, Pablo; Lizana, Angel; Peinado, Alba; Mora-González, Miguel; Campos, Juan

    2015-05-01

    A technique based on a spatial light modulator is proposed for the labeling of nonplanar surfaces. This technique arises from the combination of a general method for generation of digital holograms with a diffractive axilens. Unlike existing methods in which hologram contrast critically decreases out of the focal plane, our proposed approach allows the system to increase the focus depth, resulting in enhanced hologram images out of the focal plane. Two existing methods for hologram generation are theoretically and experimentally compared in terms of efficiency and contrast: the iterative Fourier transform algorithm-based method and the random multiplexing of different lenses method. The most suitable one is selected for the implementation of the new axilens-based technique. Experimental results obtained with the axilens method provide a significant enhancement of the image quality of holograms obtained out of the focal plane when compared with an existing technique. The approach proposed in this manuscript may be of interest in industrial applications, where alphanumeric data must be registered on nonflat surfaces, as in the cases of beverage cans or bottles.

  11. Aquifer water abundance evaluation using a fuzzy- comprehensive weighting method

    NASA Astrophysics Data System (ADS)

    Wei, Z.

    2016-08-01

    Aquifer water abundance evaluation is a highly relevant issue that has been researched for many years. Despite prior research, problems with the conventional evaluation method remain. This paper establishes an aquifer water abundance evaluation method that combines fuzzy evaluation with a comprehensive weighting method to overcome both the subjectivity and lack of conformity in determining weight by pure data analysis alone. First, this paper introduces the principle of a fuzzy-comprehensive weighting method. Second, the example of well field no. 3 (of a coalfield) is used to illustrate the method's process. The evaluation results show that this method is can more suitably meet the real requirements of aquifer water abundance assessment, leading to more precise and accurate evaluations. Ultimately, this paper provides a new method for aquifer water abundance evaluation.

  12. Whole-brain neural network analysis (connectomics) using cell lineage-based neuron-labeling method.

    PubMed

    Ito, Kei; Ito, Masayoshi

    2014-11-01

    The brain is a computing machine that receives input signals from sensory neurons, calculates best responses to changing environments, and sends output signals to motor muscles. How such computation is materialized remains largely unknown. Understanding the entire wiring network of neural connections in the brain, which is recently called the connectomics (connection + omics), should provide indispensable insights on this problem.To resolve the circuit diagram from the tangled thickets of neural fibers, only a small subset of neurons should be visualized at one time. Previous studies visualized such selective cells by injecting dyes or by detecting specific molecules or gene expression patterns using antibodies and expression driver strains. These approaches were unfortunately not efficient enough for identifying all the brain cells in a comprehensive and systematic manner.Neurons are generated by neural stem cells. The entire neural population can therefore be divided into a finite number of families - or clones - of the cells that are the progeny of each single stem cell. The central brain of the fruit fly Drosophila melanogaster consists of about 15,000 neurons per side and is made by utmost 100 stem cells. By genetically labeling one of such stem cells and tracing the projection patterns of its progeny in the adult brain, we were able to identify the neural projections of almost all the clonal cell groups.To visualize these neural projections, we made serial optical sections of the fly brain using laser confocal microscopy. Because of its relatively small size (0.6-mm wide and less than 0.3-mm thick), the entire fly brain can be imaged using high-resolution objectives with n.a. 1.2. Neuronal fibers are visualized by ectopically expressed cytoplasmic and membrane-bound fluorescent proteins, and the output synaptic sites are visualized with ectopically expressed tag proteins that are fused with the proteins associated with synaptic vesicles. In addition, density

  13. Application of improved extension evaluation method to water quality evaluation

    NASA Astrophysics Data System (ADS)

    Wong, Heung; Hu, Bao Qing

    2014-02-01

    The extension evaluation method (EEM) has been developed and applied to evaluate water quality. There are, however, negative values in the correlative degree (water quality grades from EEM) after the calculation. This is not natural as the correlative degree is essentially an index based on grades (rankings) of water quality by different methods, which are positive. To overcome this negativity issue, the interval clustering approach (ICA) was introduced, which is based on the grey clustering approach (GCA) and interval-valued fuzzy sets. However, the computing process and formulas of ICA are rather complex. This paper provides a novel method, i.e., improved extension evaluation method, so as to avoid negative values in the correlative degree. To demonstrate our proposed approach, the improved EEM is applied to evaluate the water quality of three different cross-sections of the Fen River, the second major branch river of the Yellow River in China and the Han Jiang River, one of the major branch rivers of the Yangtse River in China. The results of the improved evaluation method are basically the same as the official water quality. The proposed method possesses also the same merit as the EEM and ICA method, which can be applied to assess water quality when the levels of attributes are defined in terms of intervals in the water quality criteria. Existing methods are mostly applicable to data in the form of single numeric values.

  14. In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: the application of the enzyme-labeled antigen method

    PubMed Central

    Mizutani, Y; Tsuge, S; Takeda, H; Hasegawa, Y; Shiogama, K; Onouchi, T; Inada, K; Sawasaki, T; Tsutsumi, Y

    2014-01-01

    Porphyromonas gingivalis is a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme-labeled antigen method was applied to detecting plasma cells producing P. gingivalis-specific antibodies in biopsied gingival tissue of periodontitis. N-terminally biotinylated P. gingivalis antigens, Ag53 and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro and Lys-hgp) were prepared by the cell-free protein synthesis system using wheatgerm extract. With these five labeled proteins as probes, 20 lesions of periodontitis were evaluated. With the AlphaScreen method, antibodies against any one of the five P. gingivalis antigens were detected in 11 (55%) serum samples and 17 (85%) tissue extracts. Using the enzyme-labeled antigen method on paraformaldehyde-fixed frozen sections of gingival tissue, plasma cells were labeled with any one of the five antigens in 17 (94%) of 18 specimens, in which evaluable plasma cells were detected. The positivity rates in periodontitis were significantly higher than those found previously in radicular cysts (20% in sera and 33% in tissue extracts with the AlphaScreen method, and 25% with the enzyme-labeled antigen method). Our findings directly indicate that antibodies reactive to P. gingivalis are locally produced in the gingival lesions, and that inflammatory reactions against P. gingivalis are involved in periodontitis. PMID:24698402

  15. A faster plant stem-water extraction method.

    PubMed

    Vendramini, Patricia F; Sternberg, Leonel da S L

    2007-01-01

    Oxygen and hydrogen isotope ratios of stem water have been used by several studies which relate the ecophysiology of plants to their water source. Undoubtedly, there are several other applications and research areas which could use this type of analysis. However, the most often used methods of extracting stem water are slow, limiting the rate of sampling and consequently preventing a deeper understanding of spatial and temporal plant water source use. We have developed a faster batch method of stem-water extraction and compare it with the most commonly used online method of stem-water extraction. Samples are sealed in 18 cm long ampoules having their extremities placed sample end in a heating block and the condensing end in a cooling block, and allowed to distill overnight. Up to 72 samples can be distilled overnight and sealed the next morning. The isotope ratios of water distilled by the batch method introduced here compared with those from the online method were in excellent agreement. In addition to being faster, this method does not need the monitoring of hot water baths and liquid nitrogen traps during distillation and does not require a complex vacuum system. Copyright (c) 2006 John Wiley & Sons, Ltd.

  16. Detection of coliform organisms in drinking water by radiometric method.

    PubMed

    Khurshid, S J; Bibi, S

    1991-07-01

    The radiometric method has been used for detection of coliform bacteria in water. The method is based on measuring the released metabolic 14CO2 from 14C-lactose in growth media containing coliform organisms incubated at 37 degrees C under continuous shaking. This rapid and sensitive radiometric method permits the detection of even single coliform organisms within 6 hours of incubation. Using this automated method, a total of 102 samples (in duplicate) collected from different areas in and around Rawalpindi and Islamabad were assessed for coliform bacteria. Of these 102 samples, 50 were tap water samples, 40 from wells and 6 each were from Rawal and Simly dams. About 47% and 67% tap water samples, while 62% and 74% well water samples were found unsatisfactory from around Islamabad and Rawalpindi areas, respectively. About 83% and 66% water samples from Rawal dam and Simly dam respectively were found to be unsatisfactory.

  17. Compositions and methods for removing arsenic in water

    DOEpatents

    Gadgil, Ashok Jagannth [El Cerrito, CA

    2011-02-22

    Compositions and methods and for contaminants from water are provided. The compositions comprise ferric hydroxide and ferric oxyhydride coated substrates for use in removing the contaminant from the water. Contacting water bearing the contaminant with the substrates can substantially reduce contaminant levels therein. Methods of oxidizing the contaminants in water to facilitate their removal by the ferric hydroxide and ferric oxyhydride coated substrates are also provided. The contaminants include, but are not limited to, arsenic, selenium, uranium, lead, cadmium, nickel, copper, zinc, chromium and vanadium, their oxides and soluble salts thereof.

  18. High performance liquid chromatography-tandem mass spectrometry method for ex vivo metabolic studies of a rhenium-labeled radiopharmaceutical for liver cancer.

    PubMed

    Chen, Wei-Hsi; Liao, Chen-Wei; Luo, Tsai-Yueh; Chang, Yu; Men, Lee-Chung; Hsieh, Yi-Cheng

    2014-01-01

    The radio-isotope rhenium-labeled N-[2-(triphenylmethyl)thioethyl]-3-aza-19-ethyloxycarbonyl-3-[2-(triphenylmethyl)thioethyl] octadecanoate) ligand (188Re-MN-16ET) is a novel therapeutic agent under preclinical evaluation for hepatoma. A reversed-phase high performance liquid chromatography coupled with a tandem mass spectrometric analysis method and diode array detector (DAD) involving a T type splitter was developed to characterize this pharmaceutical in rat liver tissue solution and determine its biotransformation rate. The separation was accomplished on a C18 column (chromolith silica, 4.6 mm x 100 mm) using an acetonitrile-ammonium acetate buffer gradient as the mobile phase. The detection was achieved by DAD set at 250nm and tandem mass spectrometry using electrospray ionization in the positive ion mode. Re-MN-16ET displayed a retention time of 23.2 min and a transition ion pair corresponding to m/z677 --> 631 for multiple reaction monitoring. Its biotransformation reaction in rat liver homogenate proceeded for 90 min in a 37°C water bath. The characterization was conducted using aliquots that were extracted and concentrated from the reaction mixture for various incubation times. Re-MN-16ET exhibited a biotransformation half-life (t1/2) of 8-9 min in liver tissue solution and was almost completely exhausted after 90 min. Two of its metabolites, consisting of the Re-labeled carboxylic acid derivative, predominately, and its corresponding demetallized disulfide ligand were found in the liver homogenate, providing a metabolism pathway for the radio-pharmaceutical.

  19. Improvements to the DRASTIC ground-water vulnerability mapping method

    USGS Publications Warehouse

    Rupert, Michael G.

    1999-01-01

    Ground-water vulnerability maps are designed to show areas of greatest potential for ground-water contamination on the basis of hydrogeologic and anthropogenic (human) factors. The maps are developed by using computer mapping hardware and software called a geographic information system (GIS) to combine data layers such as land use, soils, and depth to water. Usually, ground-water vulnerability is determined by assigning point ratings to the individual data layers and then adding the point ratings together when those layers are combined into a vulnerability map. Probably the most widely used ground-water vulnerability mapping method is DRASTIC, named for the seven factors considered in the method: Depth to water, net Recharge, Aquifer media, Soil media, Topography, Impact of vadose zone media, and hydraulic Conductivity of the aquifer (Aller and others, 1985, p. iv). The DRASTIC method has been used to develop ground-water vulnerability maps in many parts of the Nation; however, the effectiveness of the method has met with mixed success (Koterba and others, 1993, p. 513; U.S. Environmental Protection Agency, 1993; Barbash and Resek, 1996; Rupert, 1997). DRASTIC maps usually are not calibrated to measured contaminant concentrations. The DRASTIC ground-water vulnerability mapping method was improved by calibrating the point rating scheme to measured nitrite plus nitrate as nitrogen (NO2+NO3–N) concentrations in ground water on the basis of statistical correlations between NO2+NO3–N concentrations and land use, soils, and depth to water (Rupert, 1997). This report describes the calibration method developed by Rupert and summarizes the improvements in results of this method over those of the uncalibrated DRASTIC method applied by Rupert and others (1991) in the eastern Snake River Plain, Idaho.

  20. Multi-label spacecraft electrical signal classification method based on DBN and random forest.

    PubMed

    Li, Ke; Yu, Nan; Li, Pengfei; Song, Shimin; Wu, Yalei; Li, Yang; Liu, Meng

    2017-01-01

    In spacecraft electrical signal characteristic data, there exists a large amount of data with high-dimensional features, a high computational complexity degree, and a low rate of identification problems, which causes great difficulty in fault diagnosis of spacecraft electronic load systems. This paper proposes a feature extraction method that is based on deep belief networks (DBN) and a classification method that is based on the random forest (RF) algorithm; The proposed algorithm mainly employs a multi-layer neural network to reduce the dimension of the original data, and then, classification is applied. Firstly, we use the method of wavelet denoising, which was used to pre-process the data. Secondly, the deep belief network is used to reduce the feature dimension and improve the rate of classification for the electrical characteristics data. Finally, we used the random forest algorithm to classify the data and comparing it with other algorithms. The experimental results show that compared with other algorithms, the proposed method shows excellent performance in terms of accuracy, computational efficiency, and stability in addressing spacecraft electrical signal data.

  1. Multi-label spacecraft electrical signal classification method based on DBN and random forest

    PubMed Central

    Li, Ke; Yu, Nan; Li, Pengfei; Song, Shimin; Wu, Yalei; Li, Yang; Liu, Meng

    2017-01-01

    In spacecraft electrical signal characteristic data, there exists a large amount of data with high-dimensional features, a high computational complexity degree, and a low rate of identification problems, which causes great difficulty in fault diagnosis of spacecraft electronic load systems. This paper proposes a feature extraction method that is based on deep belief networks (DBN) and a classification method that is based on the random forest (RF) algorithm; The proposed algorithm mainly employs a multi-layer neural network to reduce the dimension of the original data, and then, classification is applied. Firstly, we use the method of wavelet denoising, which was used to pre-process the data. Secondly, the deep belief network is used to reduce the feature dimension and improve the rate of classification for the electrical characteristics data. Finally, we used the random forest algorithm to classify the data and comparing it with other algorithms. The experimental results show that compared with other algorithms, the proposed method shows excellent performance in terms of accuracy, computational efficiency, and stability in addressing spacecraft electrical signal data. PMID:28486479

  2. Validation of an Online Food Frequency Questionnaire against Doubly Labelled Water and 24 h Dietary Recalls in Pre-School Children

    PubMed Central

    Delisle Nyström, Christine; Henriksson, Hanna; Alexandrou, Christina; Bergström, Anna; Bonn, Stephanie; Bälter, Katarina; Löf, Marie

    2017-01-01

    The development of easy-to-use and accurate methods to assess the intake of energy, foods and nutrients in pre-school children is needed. KidMeal-Q is an online food frequency questionnaire developed for the LifeGene prospective cohort study in Sweden. The aims of this study were to compare: (i) energy intake (EI) obtained using KidMeal-Q to total energy expenditure (TEE) measured via doubly labelled water and (ii) the intake of certain foods measured using KidMeal-Q to intakes acquired by means of 24 h dietary recalls in 38 children aged 5.5 years. The mean EI calculated using KidMeal-Q was statistically different (p < 0.001) from TEE (4670 ± 1430 kJ/24 h and 6070 ± 690 kJ/24 h, respectively). Significant correlations were observed for vegetables, fruit juice and candy between KidMeal-Q and 24 h dietary recalls. Only sweetened beverage consumption was significantly different in mean intake (p < 0.001), as measured by KidMeal-Q and 24 h dietary recalls. In conclusion, KidMeal-Q had a relatively short answering time and comparative validity to other food frequency questionnaires. However, its accuracy needs to be improved before it can be used in studies in pre-school children. PMID:28098765

  3. Nanostructured luminescently labeled nucleic acids.

    PubMed

    Kricka, Larry J; Fortina, Paolo; Park, Jason Y

    2017-03-01

    Important and emerging trends at the interface of luminescence, nucleic acids and nanotechnology are: (i) the conventional luminescence labeling of nucleic acid nanostructures (e.g. DNA tetrahedron); (ii) the labeling of bulk nucleic acids (e.g. single-stranded DNA, double-stranded DNA) with nanostructured luminescent labels (e.g. copper nanoclusters); and (iii) the labeling of nucleic acid nanostructures (e.g. origami DNA) with nanostructured luminescent labels (e.g. silver nanoclusters). This review surveys recent advances in these three different approaches to the generation of nanostructured luminescently labeled nucleic acids, and includes both direct and indirect labeling methods. Copyright © 2016 John Wiley & Sons, Ltd.

  4. Pharmacokinetics study of calf thymus DNA in rats and beagle dogs with (3)H-labeling and tracing method.

    PubMed

    Yang, Shuoye; Talbi, Amer; Wang, Xin; Song, Hanlin; Chen, Xijing

    2014-01-01

    This study developed a radioisotope detection and tracing method to investigate the pharmacokinetic properties of calf thymus DNA (ctDNA) in rats and beagle dogs. The radioactivity labeling result was detected through gel electrophoresis analysis, and pharmacokinetic analytical methods for (3)H-ctDNA in rat and beagle dog plasma were developed, respectively. Full method validation indicated that the established radioisotope method was sensitive, specific, rapid and reliable, and the results were all in accordance with the analysis requirement in biological samples. After intravenous administration of the planned doses of (3)H-ctDNA to the rats and beagle dogs, plasma concentrations from the various dose groups declined rapidly. In addition, the radioactive concentration of (3)H-ctDNA in the plasma from single and multiple dosings decreased in a similar trend. Through comparative analysis of the pharmacokinetic parameters, we inferred that the elimination of ctDNA accorded with the linear pharmacokinetic characteristic. The results demonstrated that ctDNA was rapidly eliminated in rat and beagle dog plasma and would not accumulate, indicating the safe use of ctDNA as an immunoadsorptive material without bringing out potential risk.

  5. SWeRF--A method for estimating the relevant fine particle fraction in bulk materials for classification and labelling purposes.

    PubMed

    Pensis, Ingeborg; Luetzenkirchen, Frank; Friede, Bernd

    2014-05-01

    In accordance with the European regulation for classification, labelling and packaging of substances and mixtures (CLP) as well as the criteria as set out in the Globally Harmonized System (GHS), fine fraction of crystalline silica (CS) has been classified as a specific target organ toxicity, the specific organ in this case being the lung. Generic cut-off values for products containing a fine fraction of CS trigger the need for a method for the quantification of the fine fraction of CS in bulk materials. This article describes the so-called SWeRF method, the size-weighted relevant fine fraction. The SWeRF method combines the particle size distribution of a powder with probability factors from the EN 481 standard and allows the relevant fine fraction of a material to be calculated. The SWeRF method has been validated with a number of industrial minerals. This will enable manufacturers and blenders to apply the CLP and GHS criteria for the classification of mineral products containing RCS a fine fraction of CS.

  6. SWeRF—A Method for Estimating the Relevant Fine Particle Fraction in Bulk Materials for Classification and Labelling Purposes

    PubMed Central

    2014-01-01

    In accordance with the European regulation for classification, labelling and packaging of substances and mixtures (CLP) as well as the criteria as set out in the Globally Harmonized System (GHS), fine fraction of crystalline silica (CS) has been classified as a specific target organ toxicity, the specific organ in this case being the lung. Generic cut-off values for products containing a fine fraction of CS trigger the need for a method for the quantification of the fine fraction of CS in bulk materials. This article describes the so-called SWeRF method, the size-weighted relevant fine fraction. The SWeRF method combines the particle size distribution of a powder with probability factors from the EN 481 standard and allows the relevant fine fraction of a material to be calculated. The SWeRF method has been validated with a number of industrial minerals. This will enable manufacturers and blenders to apply the CLP and GHS criteria for the classification of mineral products containing RCS a fine fraction of CS. PMID:24389081

  7. Calibration of the DRASTIC ground water vulnerability mapping method

    USGS Publications Warehouse

    Rupert, M.G.

    2001-01-01

    Ground water vulnerability maps developed using the DRASTIC method have been produced in many parts of the world. Comparisons of those maps with actual ground water quality data have shown that the DRASTIC method is typically a poor predictor of ground water contamination. This study significantly improved the effectiveness of a modified DRASTIC ground water vulnerability map by calibrating the point rating schemes to actual ground water quality data by using nonparametric statistical techniques and a geographic information system. Calibration was performed by comparing data on nitrite plus nitrate as nitrogen (NO2 + NO3-N) concentrations in ground water to land-use, soils, and depth to first-encountered ground water data. These comparisons showed clear statistical differences between NO2 + NO3-N concentrations and the various categories. Ground water probability point ratings for NO2 + NO3-N contamination were developed from the results of these comparisons, and a probability map was produced. This ground water probability map was then correlated with an independent set of NO2 + NO3-N data to demonstrate its effectiveness in predicting elevated NO2 + NO3-N concentrations in ground water. This correlation demonstrated that the probability map was effective, but a vulnerability map produced with the uncalibrated DRASTIC method in the same area and using the same data layers was not effective. Considerable time and expense have been outlaid to develop ground water vulnerability maps with the DRASTIC method. This study demonstrates a cost-effective method to improve and verify the effectiveness of ground water vulnerability maps.

  8. A facile method for expression and purification of 15N isotope-labeled human Alzheimer's β-amyloid peptides from E. coli for NMR-based structural analysis

    PubMed Central

    Armand, Tara; Ball, K. Aurelia; Chen, Anna; Pelton, Jeffrey G.; Wemmer, David E.; Head-Gordon, Teresa

    2016-01-01

    Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting millions of people worldwide. AD is characterized by the presence of extracellular plaques composed of aggregated/oligomerized β-amyloid peptides with Aβ42 peptide representing a major isoform in the senile plaques. Given the pathological significance of Aβ42 in the progression of AD, there is considerable interest in understanding the structural ensembles for soluble monomer and oligomeric forms of Aβ42. This report describes an efficient method to express and purify high quality 15N isotope-labeled Aβ42 for structural studies by NMR. The protocol involves utilization of an auto induction system with 15N isotope labeled medium, for high-level expression of Aβ42 as a fusion with IFABP. After the over-expression of the 15N isotope-labeled IFABP-Aβ42 fusion protein in the inclusion bodies, pure 15N isotope-labeled Aβ42 peptide is obtained following a purification method that is streamlined and improved from the method originally developed for the isolation of unlabeled Aβ42 peptide (Garai et al., 2009). We obtain a final yield of ∼6 mg/L culture for 15N isotope-labeled Aβ42 peptide. Mass spectrometry and 1H–15N HSQC spectra of monomeric Aβ42 peptide validate the uniform incorporation of the isotopic label. The method described here is equally applicable for the uniform isotope labeling with 15N and 13C in Aβ42 peptide as well as its other variants including any Aβ42 peptide mutants. PMID:26231074

  9. Dual Labeling Biotin Switch Assay to Reduce Bias Derived from Different Cysteine Subpopulations: A Method to Maximize S-Nitrosylation Detection

    PubMed Central

    Chung, Heaseung Sophia; Murray, Christopher I.; Venkatraman, Vidya; Crowgey, Erin L.; Rainer, Peter P.; Cole, Robert N.; Bomgarden, Ryan D.; Rogers, John C.; Balkan, Wayne; Hare, Joshua M.; Kass, David A.; Van Eyk, Jennifer E.

    2016-01-01

    Rationale S-nitrosylation (SNO), an oxidative post-translational modification of cysteine residues, responds to changes in the cardiac redox-environment. Classic biotin switch assay and its derivatives are the most common methods used for detecting SNO. In this approach, the labile SNO group is selectively replaced with a single stable tag. To date, a variety of thiol-reactive tags have been introduced. However, these methods have not produced a consistent dataset which suggests an incomplete capture by a single tag and potentially the presence of different cysteine subpopulations. Objective To investigate potential labeling bias in the existing methods with a single tag to detect SNO, explore if there are distinct cysteine subpopulations, and then, develop a strategy to maximize the coverage of SNO proteome. Methods and Results We obtained SNO-modified cysteine datasets for wild-type and S-nitrosoglutathione reductase (GSNOR) knock-out mouse hearts (GSNOR is a negative regulator of GSNO production) and NO-induced human embryonic kidney cell using two labeling reagents; the cysteine-reactive pyridyldithiol and iodoacetyl based tandem mass tags. Comparison revealed that <30% of the SNO-modified residues were detected by both tags, while the remaining SNO sites were only labeled by one reagent. Characterization of the two distinct subpopulations of SNO residues indicated that pyridyldithiol reagent preferentially labels cysteine residues that are more basic and hydrophobic. Based on this observation, we proposed a parallel dual labeling strategy followed by an optimized proteomics workflow. This enabled the profiling of 493 SNO-sites in GSNOR knock-out hearts. Conclusions Using a protocol comprising two tags for dual labeling maximizes overall detection of SNO by reducing the previously unrecognized labeling bias derived from different cysteine subpopulations. PMID:26338901

  10. Extending electromagnetic methods to map coastal pore water salinities

    USGS Publications Warehouse

    Greenwood, Wm. J.; Kruse, S.; Swarzenski, P.

    2006-01-01

    The feasibility of mapping pore water salinity based on surface electromagnetic (EM) methods over land and shallow marine water is examined in a coastal wetland on Tampa Bay, Florida. Forward models predict that useful information on seabed conductivity can be obtained through <1.5 m of saline water, using floating EM-31 and EM-34 instruments from Geonics Ltd. The EM-31 functioned as predicted when compared against resistivity soundings and pore water samples and proved valuable for profiling in otherwise inaccessible terrain due to its relatively small size. Experiments with the EM-34 in marine water, however, did not reproduce the theoretical instrument response. The most effective technique for predicting pore water conductivities based on EM data entailed (1) computing formation factors from resistivity surveys and pore water samples at representative sites and (2) combining these formation factors with onshore and offshore EM-31 readings for broader spatial coverage. This method proved successful for imaging zones of elevated pore water conductivities/ salinities associated with mangrove forests, presumably caused by salt water exclusion by mangrove roots. These zones extend 5 to 10 m seaward from mangrove trunks fringing Tampa Bay. Modeling indicates that EM-31 measurements lack the resolution necessary to image the subtle pore water conductivity variations expected in association with diffuse submarine ground water discharge of fresher water in the marine water of Tampa Bay. The technique has potential for locating high-contrast zones and other pore water salinity anomalies in areas not accessible to conventional marine- or land-based resistivity arrays and hence may be useful for studies of coastal-wetland ecosystems. Copyright ?? 2005 National Ground Water Association.

  11. An Image Analysis Method for the Precise Selection and Quantitation of Fluorescently Labeled Cellular Constituents

    PubMed Central

    Agley, Chibeza C.; Velloso, Cristiana P.; Lazarus, Norman R.

    2012-01-01

    The accurate measurement of the morphological characteristics of cells with nonuniform conformations presents difficulties. We report here a straightforward method using immunofluorescent staining and the commercially available imaging program Adobe Photoshop, which allows objective and precise information to be gathered on irregularly shaped cells. We have applied this measurement technique to the analysis of human muscle cells and their immunologically marked intracellular constituents, as these cells are prone to adopting a highly branched phenotype in culture. Use of this method can be used to overcome many of the long-standing limitations of conventional approaches for quantifying muscle cell size in vitro. In addition, wider applications of Photoshop as a quantitative and semiquantitative tool in immunocytochemistry are explored. PMID:22511600

  12. SEC-TID: A Label-Free Method for Small-Molecule Target Identification.

    PubMed

    Salcius, Michael; Bauer, Andras J; Hao, Qin; Li, Shu; Tutter, Antonin; Raphael, Jacob; Jahnke, Wolfgang; Rondeau, Jean-Michel; Bourgier, Emmanuelle; Tallarico, John; Michaud, Gregory A

    2014-07-01

    Bioactive small molecules are an invaluable source of therapeutics and chemical probes for exploring biological pathways. Yet, significant hurdles in drug discovery often come from lacking a comprehensive view of the target(s) for both early tool molecules and even late-stage drugs. To address this challenge, a method is provided that allows for assessing the interactions of small molecules with thousands of targets without any need to modify the small molecule of interest or attach any component to a surface. We describe size-exclusion chromatography for target identification (SEC-TID), a method for accurately and reproducibly detecting ligand-macromolecular interactions for small molecules targeting nucleic acid and several protein classes. We report the use of SEC-TID, with a library consisting of approximately 1000 purified proteins derived from the protein databank (PDB), to identify the efficacy targets tankyrase 1 and 2 for the Wnt inhibitor XAV939. In addition, we report novel interactions for the tumor-vascular disrupting agent vadimezan/ASA404 (interacting with farnesyl pyrophosphate synthase) and the diuretic mefruside (interacting with carbonic anhydrase XIII). We believe this method can dramatically enhance our understanding of the mechanism of action and potential liabilities for small molecules in drug discovery pipelines through comprehensive profiling of candidate druggable targets.

  13. Chemical Methods for the Direct Detection and Labeling of S-Nitrosothiols

    PubMed Central

    Bechtold, Erika

    2012-01-01

    Abstract Significance: Posttranslational modification of proteins through phosphorylation, glycosylation, and oxidation adds complexity to the proteome by reversibly altering the structure and function of target proteins in a highly controlled fashion. Recent Advances: The study of reversible cysteine oxidation highlights a role for this oxidative modification in complex signal transduction pathways. Nitric oxide (NO), and its respective metabolites (including reactive nitrogen species), participates in a variety of these cellular redox processes, including the reversible oxidation of cysteine to S-nitrosothiols (RSNOs). RSNOs act as endogenous transporters of NO, but also possess beneficial effects independent of NO-related signaling, which suggests a complex and versatile biological role. In this review, we highlight the importance of RSNOs as a required posttranslational modification and summarize the current methods available for detecting S-nitrosation. Critical Issues: Given the limitations of these indirect detection methods, the review covers recent developments toward the direct detection of RSNOs by phosphine-based chemical probes. The intrinsic properties that dictate this phosphine/RSNO reactivity are summarized. In general, RSNOs (both small molecule and protein) react with phosphines to yield reactive S-substituted aza-ylides that undergo further reactions leading to stable RSNO-based adducts. Future Directions: This newly explored chemical reactivity forms the basis of a number of exciting potential chemical methods for protein RSNO detection in biological systems. Antioxid. Redox Signal. 17, 981–991. PMID:22356122

  14. Detection of uracil within DNA using a sensitive labeling method for in vitro and cellular applications

    PubMed Central

    Róna, Gergely; Scheer, Ildikó; Nagy, Kinga; Pálinkás, Hajnalka L.; Tihanyi, Gergely; Borsos, Máté; Békési, Angéla; Vértessy, Beáta G.

    2016-01-01

    The role of uracil in genomic DNA has been recently re-evaluated. It is now widely accepted to be a physiologically important DNA element in diverse systems from specific phages to antibody maturation and Drosophila development. Further relevant investigations would largely benefit from a novel reliable and fast method to gain quantitative and qualitative information on uracil levels in DNA both in vitro and in situ, especially since current techniques does not allow in situ cellular detection. Here, starting from a catalytically inactive uracil-DNA glycosylase protein, we have designed several uracil sensor fusion proteins. The designed constructs can be applied as molecular recognition tools that can be detected with conventional antibodies in dot-blot applications and may also serve as in situ uracil-DNA sensors in cellular techniques. Our method is verified on numerous prokaryotic and eukaryotic cellular systems. The method is easy to use and can be applied in a high-throughput manner. It does not require expensive equipment or complex know-how, facilitating its easy implementation in any basic molecular biology laboratory. Elevated genomic uracil levels from cells of diverse genetic backgrounds and/or treated with different drugs can be demonstrated also in situ, within the cell. PMID:26429970

  15. A Highly Selective and Sensitive Fluorescence Detection Method of Glyphosate Based on an Immune Reaction Strategy of Carbon Dot Labeled Antibody and Antigen Magnetic Beads.

    PubMed

    Wang, Duo; Lin, Bixia; Cao, Yujuan; Guo, Manli; Yu, Ying

    2016-08-03

    A sensitive fluorescence detection method for glyphosate (GLY) was established based on immune reaction. First, carbon dot labeled antibodies (lgG-CDs) which were able to specifically identify glyphosate were prepared with the environmentally friendly carbon dots (CDs) and glyphosate antibody (lgG). lgG-CDs could be used to in situ visualize the distribution of glyphosate in plant tissues. In order to eliminate the effects of excess lgG-CDs on the determination of GLY, antigen magnetic beads Fe3O4-GLY based on magnetic nanoparticles Fe3O4 and glyphosate were constructed and utilized to couple with the excess lgG-CDs. After magnetic separation to remove antigen magnetic beads, there was a linear relationship between the fluorescence intensity of lgG-CDs and the logarithmic concentration of glyphosate in the range of 0.01-80 μg/mL with a detection limit of 8 ng/mL. The method was used for the detection of glyphosate in Pearl River water, tea, and soil samples with satisfactory recovery ratio between 87.4% and 103.7%.

  16. Indium-111-labeled autologous leukocyte imaging and fecal excretion. Comparison with conventional methods of assessment of inflammatory bowel disease

    SciTech Connect

    Leddin, D.J.; Paterson, W.G.; DaCosta, L.R.; Dinda, P.K.; Depew, W.T.; Markotich, J.; McKaigney, J.P.; Groll, A.; Beck, I.T.

    1987-04-01

    This study was designed to evaluate the role of /sup 111/In-labeled leukocyte imaging and fecal excretion in the assessment of inflammatory bowel disease. We compared these tests to various indices of disease activity in Crohn's disease, to Truelove's grading in ulcerative colitis, and to endoscopy, x-ray, and pathology in both diseases. Eleven controls, 16 patients with Crohn's disease, 13 with ulcerative colitis, and 3 with other types of acute bowel inflammation were studied (positive controls). Indium scanning was performed at 1, 4, and 24 hr. Fourteen of 16 patients with active Crohn's disease had positive scans but in only five was localization accurate. One patient had inactive ulcerative colitis, and the scan was negative. Of 12 patients with active ulcerative colitis, 10 had positive scans but disease localization was accurate in only four. Disease extent was correctly defined in 1 of the 3 Positive Controls. There was no significant difference in the accuracy of scanning at 1, 4, or 24 hr. /sup 111/In fecal excretion was significantly higher in patients with inflammatory bowel disease than in controls, and there was correlation between /sup 111/In fecal excretion and most of the indices of disease activity in Crohn's disease. In ulcerative colitis, /sup 111/In fecal excretion did not correlate with Truelove's grading but reflected colonoscopic assessment of severity. In conclusion, /sup 111/In-labeled leukocyte scanning lacks sensitivity with respect to disease extent, but fecal excretion of /sup 111/In correlates well with disease severity as determined by other methods.

  17. Waste Water Treatment Apparatus and Methods

    NASA Technical Reports Server (NTRS)

    Littman, Howard (Inventor); Plawsky, Joel L. (Inventor); Paccione, John D. (Inventor)

    2014-01-01

    An improved draft tube spout fluid bed (DTSFB) mixing, handling, conveying, and treating apparatus and systems, and methods for operating are provided. The apparatus and systems can accept particulate material and pneumatically or hydraulically conveying the material to mix and/or treat the material. In addition to conveying apparatus, a collection and separation apparatus adapted to receive the conveyed particulate material is also provided. The collection apparatus may include an impaction plate against which the conveyed material is directed to improve mixing and/or treatment. The improved apparatus are characterized by means of controlling the operation of the pneumatic or hydraulic transfer to enhance the mixing and/or reacting by controlling the flow of fluids, for example, air, into and out of the apparatus. The disclosed apparatus may be used to mix particulate material, for example, mortar; react fluids with particulate material; coat particulate material, or simply convey particulate material.

  18. ISO standards on test methods for water radioactivity monitoring.

    PubMed

    Calmet, D; Ameon, R; Bombard, A; Forte, M; Fournier, M; Herranz, M; Jerome, S; Kwakman, P; Llaurado, M; Tokonami, S

    2013-11-01

    Water is vital to humans and each of us needs at least 1.5L of safe water a day to drink. Beginning as long ago as 1958 the World Health Organization (WHO) has published guidelines to help ensure water is safe to drink. Focused from the start on monitoring radionuclides in water, and continually cooperating with WHO, the International Standardization Organization (ISO) has been publishing standards on radioactivity test methods since 1978. As reliable, comparable and 'fit for purpose' results are an essential requirement for any public health decision based on radioactivity measurements, international standards of tested and validated radionuclide test methods are an important tool for production of such measurements. This paper presents the ISO standards already published that could be used as normative references by testing laboratories in charge of radioactivity monitoring of drinking water as well as those currently under drafting and the prospect of standardized fast test methods in response to a nuclear accident.

  19. Rapid and label-free bioanalytical method of alpha fetoprotein detection using LSPR chip

    NASA Astrophysics Data System (ADS)

    Kim, Dongjoo; Kim, Jinwoon; Kwak, Cheol Hwan; Heo, Nam Su; Oh, Seo Yeong; Lee, Hoomin; Lee, Go-Woon; Vilian, A. T. Ezhil; Han, Young-Kyu; Kim, Woo-Sik; Kim, Gi-bum; Kwon, Soonjo; Huh, Yun Suk

    2017-07-01

    Alpha fetoprotein (AFP) is a cancer marker, particularly for hepatocellular carcinoma. Normal levels of AFP are less than 20 ng/mL; however, its levels can reach more than 400 ng/mL in patients with HCC. Enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) have been employed for clinical diagnosis of AFP; however, these methods are time consuming and labor intensive. In this study, we developed a localized surface plasmon resonance (LSPR) based biosensor for simple and rapid detection of AFP. This biosensor consists of a UV-Vis spectrometer, a cuvette cell, and a biosensor chip nanopatterned with gold nanoparticles (AuNPs). In our LSPR biosensor, binding of AFP to the surface of the sensor chip led to an increasing magnitude of the LSPR signals, which was measured by an ultraviolet-visible (UV-Vis) spectrometer. Our LSPR biosensor showed sufficient detectability of AFP at concentrations of 1 ng/mL to 1 μg/mL. Moreover, the overall procedure for detection of AFP was completed within 20 min. This biosensor could also be utilized for a point of care test (POCT) by employing a portable UV-Vis spectrometer. Owing to the simplicity and rapidity of the detection process, our LSPR biosensor is expected to replace traditional diagnostic methods for the early detection of diseases.

  20. QPROT: Statistical method for testing differential expression using protein-level intensity data in label-free quantitative proteomics.

    PubMed

    Choi, Hyungwon; Kim, Sinae; Fermin, Damian; Tsou, Chih-Chiang; Nesvizhskii, Alexey I

    2015-11-03

    We introduce QPROT, a statistical framework and computational tool for differential protein expression analysis using protein intensity data. QPROT is an extension of the QSPEC suite, originally developed for spectral count data, adapted for the analysis using continuously measured protein-level intensity data. QPROT offers a new intensity normalization procedure and model-based differential expression analysis, both of which account for missing data. Determination of differential expression of each protein is based on the standardized Z-statistic based on the posterior distribution of the log fold change parameter, guided by the false discovery rate estimated by a well-known Empirical Bayes method. We evaluated the classification performance of QPROT using the quantification calibration data from the clinical proteomic technology assessment for cancer (CPTAC) study and a recently published Escherichia coli benchmark dataset, with evaluation of FDR accuracy in the latter. QPROT is a statistical framework with computational software tool for comparative quantitative proteomics analysis. It features various extensions of QSPEC method originally built for spectral count data analysis, including probabilistic treatment of missing values in protein intensity data. With the increasing popularity of label-free quantitative proteomics data, the proposed method and accompanying software suite will be immediately useful for many proteomics laboratories. This article is part of a Special Issue entitled: Computational Proteomics. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Improved methods for national water assessment, water resources contract: WR15249270

    USGS Publications Warehouse

    Thomas, Harold A.

    1981-01-01

    The purpose of our research is to develop methods to make National Water Assessment more useful in estimating water availability for economic growth and more helpful in determining the effect of water resource development upon the environmental quality of related land resources. There are serious questions pertaining to the 1975 Water Assessment and these amplify the significance of decisions made as to the planning and scheduling of the next assessment.

  2. [Three quantitative methods to continuously monitor Legionella in spring water].

    PubMed

    Yan, Ge-bin; Wang, Huan-xin; Qin, Tian; Zhou, Hai-jian; Li, Ma-chao; Xu, Ying; Zhao, Ming-qiang; Shao, Zhu-jun; Ren, Hong-yu

    2013-07-01

    To compare the detection effect of Legionella pollution in spring water by three methods, namely traditional plating method, fluorescent quantitation PCR method and ethidium monoazide (EMA) fluorescent quantitation PCR method. Every month (except May), we collected 11 water samples from the 5 selected hot spring pools in one hot spring resort in Beijing in 2011. A total of 121 water samples were collected, and then were detected by the above three methods qualitatively and quantitatively. In our study, the Legionella pollution rate was separately 74.4% (90/121), 100.0% (121/121) and 100.0% (121/121) by the above three methods. The quantitative value of Legionella in the 121 water samples detected by the three methods were around 0.10-216.00 colony-forming units (CFU)/ml, 1.47-1557.75 gene units (GU)/ml and 0.20-301.69 GU/ml, respectively. The median (25th and 75th percentiles) was 75.30 (32.51-192.10) GU/ml, 36.46 (16.08-91.21) GU/ml and 5.30 (0.00-33.70) CFU/ml, respectively. The difference in the quantitative value of Legionella detected by the three methods showed statistical significance (χ(2) = 187.900, P < 0.01). The quantitative value of Legionella detected by fluorescent quantitation PCR method was the highest, followed by the value Legionella detected by EMA-fluorescent quantitation PCR method and traditional plating method. The sensitivity of the PCR methods was higher than traditional plating method, in detecting Legionella pollution in spring water, especially the EMA- fluorescent quantitation PCR method, which was more suitable for detecting Legionella in water.

  3. Ranking filter methods for concentrating pathogens in lake water

    USDA-ARS?s Scientific Manuscript database

    Accurately comparing filtration methods for concentrating waterborne pathogens is difficult because of two important water matrix effects on recovery measurements, the effect on PCR quantification and the effect on filter performance. Regarding the first effect, we show how to create a control water...

  4. River Pollution: Part II. Biological Methods for Assessing Water Quality.

    ERIC Educational Resources Information Center

    Openshaw, Peter

    1984-01-01

    Discusses methods used in the biological assessment of river quality and such indicators of clean and polluted waters as the Trent Biotic Index, Chandler Score System, and species diversity indexes. Includes a summary of a river classification scheme based on quality criteria related to water use. (JN)

  5. River Pollution: Part II. Biological Methods for Assessing Water Quality.

    ERIC Educational Resources Information Center

    Openshaw, Peter

    1984-01-01

    Discusses methods used in the biological assessment of river quality and such indicators of clean and polluted waters as the Trent Biotic Index, Chandler Score System, and species diversity indexes. Includes a summary of a river classification scheme based on quality criteria related to water use. (JN)

  6. Multi-Isotope Secondary Ion Mass Spectrometry Combining Heavy Water 2H with 15N Labeling As Complementary Tracers for Metabolic Heterogeneity at the Single-Cell Level

    NASA Astrophysics Data System (ADS)

    Kopf, S.; McGlynn, S.; Cowley, E.; Green, A.; Newman, D. K.; Orphan, V. J.

    2014-12-01

    Metabolic rates of microbial communities constitute a key physiological parameter for understanding the in situ growth constraints for life in any environment. Isotope labeling techniques provide a powerful approach for measuring such biological activity, due to the use of isotopically enriched substrate tracers whose incorporation into biological materials can be detected with high sensitivity by isotope-ratio mass spectrometry. Nano-meter scale secondary ion mass spectrometry (NanoSIMS) combined with stable isotope labeling provides a unique tool for studying the spatiometabolic activity of microbial populations at the single cell level in order to assess both community structure and population diversity. However, assessing the distribution and range of microbial activity in complex environmental systems with slow-growing organisms, diverse carbon and nitrogen sources, or heterotrophic subpopulations poses a tremendous technical challenge because the introduction of isotopically labeled substrates frequently changes the nutrient availability and can inflate or bias measures of activity. Here, we present the use of hydrogen isotope labeling with deuterated water as an important new addition to the isotopic toolkit and apply it for the determination of single cell microbial activities by NanoSIMS imaging. This tool provides a labeling technique that minimally alters any aquatic chemical environment, can be administered with strong labels even in minimal addition (natural background is very low), is an equally universal substrate for all forms of life even in complex, carbon and nitrogen saturated systems, and can be combined with other isotopic tracers. The combination of heavy water labeling with the most commonly used NanoSIMS tracer, 15N, is technically challenging but opens up a powerful new set of multi-tracer experiments for the study of microbial activity in complex communities. We present the first truly simultaneous single cell triple isotope system

  7. A rapid and sensitive GC-MS/MS method to measure deuterium labeled deoxyadenosine in DNA from limited mouse cell populations

    PubMed Central

    Farthing, Don E.; Buxbaum, Nataliya P.; Bare, Catherine V.; Treadwell, Shirin M.; Kapoor, Veena; Williams, Kirsten M.; Gress, Ronald E.

    2013-01-01

    A rapid and sensitive GC-MS/MS method was developed to quantitatively measure low levels of DNA base deoxyadenosine (dA) and its isotopologues (e.g. dA M+1) from limited mouse cell populations. Mice undergoing allogeneic hematopoietic transplantation (AHSCT) received deuterated water at biologically relevant time intervals post AHSCT, allowing labeling of DNA upon cell division, which was detected as the dA M+1 isotopologue. Targeted mouse cell populations were isolated from lymphoid organs and purified by multi-parameter fluorescence activated cell sorting. Cell lysis, DNA extraction and hydrolysis were accomplished using available commercial procedures. The novel analytical method utilized a hydrophilic-lipophilic balanced sample preparation, rapid on-line hot GC inlet gas phase sample derivatization, fast GC low thermal mass technology, and a recently marketed GC-MS/MS system. Calibration standards containing dA and fortified with relevant levels of dA M+1 (0.25–20%) and dA M+5 (internal standard) were used for sample quantitation. The method employed a quadratic fit for calibration of dA M+1 (0.25–20%) and dA, demonstrated excellent accuracy and precision, and had limits of detection of 100 fg on-column for the dA isotopologues. The method was validated and required only 20,000 cells to characterize population dynamics of cells involved in the biology of chronic graft-versus-host disease, the main cause of late morbidity and non-relapse-mortality following AHSCT. The high sensitivity and specificity of the method makes it useful for investigating in vivo kinetics on limited and important cell populations (e.g. T regulatory cells) from disease conditions or in disease models that are immune-mediated, such as diabetes, HIV/AIDS, arthritis, inflammatory bowel disease, and multiple sclerosis. PMID:23541182

  8. AN IMPROVED METHOD FOR DETECTING VIRUSES IN WATER

    EPA Science Inventory

    Enteroviruses are important etiological agents of waterborne disease and are responsible for outbreaks of gastroenteritis. However, the prevalence and occurrence of these pathogens in raw drinking water sources is poorly understood. This is primarily due to the limited methods ...

  9. AN IMPROVED METHOD FOR DETECTING VIRUSES IN WATER

    EPA Science Inventory

    Enteroviruses are important etiological agents of waterborne disease and are responsible for outbreaks of gastroenteritis. However, the prevalence and occurrence of these pathogens in raw drinking water sources is poorly understood. This is primarily due to the limited methods ...

  10. REMOVAL OF URANIUM FROM DRINKING WATER BY CONVENTIONAL TREATMENT METHODS

    EPA Science Inventory

    The USEPA currently does not regulate uranium in drinking water but will be revising the radionuclide regulations during 1989 and will propose a maximum contaminant level for uranium. The paper presents treatment technology information on the effectiveness of conventional method...

  11. REMOVAL OF URANIUM FROM DRINKING WATER BY CONVENTIONAL TREATMENT METHODS

    EPA Science Inventory

    The USEPA currently does not regulate uranium in drinking water but will be revising the radionuclide regulations during 1989 and will propose a maximum contaminant level for uranium. The paper presents treatment technology information on the effectiveness of conventional method...

  12. A bootstrap method for estimating uncertainty of water quality trends

    USGS Publications Warehouse

    Hirsch, Robert M.; Archfield, Stacey A.; DeCicco, Laura

    2015-01-01

    Estimation of the direction and magnitude of trends in surface water quality remains a problem of great scientific and practical interest. The Weighted Regressions on Time, Discharge, and Season (WRTDS) method was recently introduced as an exploratory data analysis tool to provide flexible and robust estimates of water quality trends. This paper enhances the WRTDS method through the introduction of the WRTDS Bootstrap Test (WBT), an extension of WRTDS that quantifies the uncertainty in WRTDS-estimates of water quality trends and offers various ways to visualize and communicate these uncertainties. Monte Carlo experiments are applied to estimate the Type I error probabilities for this method. WBT is compared to other water-quality trend-testing methods appropriate for data sets of one to three decades in length with sampling frequencies of 6–24 observations per year. The software to conduct the test is in the EGRETci R-package.

  13. New Remote Sensing Methods for Labeling Disturbance Agents in Appalachian Forests

    NASA Astrophysics Data System (ADS)

    Hughes, M. J.; Hayes, D. J.

    2014-12-01

    Forests in the eastern United States are species rich and affected by a variety of disturbance agents such as fire, invasive insects, diseases, and storm events. Millions of hectares of forest are disturbed each year, altering the forest carbon sink and changing forest nutrient cycles. The magnitude and direction of these changes, though, can be different for different disturbance agents. For example, trees that burn in severe fire rapidly release stored carbon into the atmosphere whereas standing deadwood from insect attacks decompose slowly while atmospheric carbon is fixed in regenerating vegetation. The diagnosis and attribution of these processes require accurate and reliable estimates of the extent and frequency of different disturbance agents. Here, a new method is presented that classifies disturbance events identified using time-series analysis of Landsat TM imagery. The method exploits information about changes in the canopy heterogeneity as measured by several texture indices within forest patches. Classifiers were trained using data from the US Forest Service Aerial Detection Surveys and currently differentiate between fires, southern pine beetle, gypsy moth, hemlock woolly adelgid, beech bark disease, anthracnose, and storm events. In addition, the classifier returns a value of 'uncertain' when it is unable to make a clear determination, which is currently approximately 10% of identified disturbances. Classification accuracy for the remainder is 81%, though is variable between agents. For example, the classifier performs well in identifying southern pine beetle and gypsy moth affected areas, but poorly in identifying storms. Reliabilities are similar to accuracies for each agent. The results presented are the first yearly, regional-scale estimates of forest disturbance partitioned by disturbance agent. We find good correspondence with previously described patterns of disturbance and distribution, including direct observational evidence of their

  14. Detection method for avian influenza viruses in water.

    PubMed

    Rönnqvist, Maria; Ziegler, Thedi; von Bonsdorff, Carl-Henrik; Maunula, Leena

    2012-03-01

    Recent events have shown that humans may become infected with some pathogenic avian influenza A viruses (AIV). Since soil and water, including lakes, rivers, and seashores, may be contaminated by AIV excreted by birds, effective methods are needed for monitoring water for emerging viruses. Combining water filtration with molecular methods such as PCR is a fast and effective way for detecting viruses. The objective of this study was to apply a convenient method for the detection of AIV in natural water samples. Distilled water and lake, river, and seawater were artificially contaminated with AIV (H5N3) and passed through a filter system. AIV was detected from filter membrane by real-time RT-PCR. The performance of Zetapor, SMWP, and Sartobind D5F membranes in recovering influenza viruses was first evaluated using contaminated distilled water. SWMP, which gave the highest virus recoveries, was then compared with a pre-filter combined GF/F filter membrane in a trial using natural water samples. In this study, the cellulose membrane SMWP was found to be practical for recovery of AIVs in water. Viral yields varied between 62.1 and 65.9% in distilled water and between 1 and 16.7% in natural water samples. The borosilicate glass membrane GF/F combined with pre-filter was also feasible in filtering natural water samples with viral yields from 1.98 to 7.33%. The methods described can be used for monitoring fresh and seawater samples for the presence of AIV and to determine the source of AIV transmission in an outbreak situation.

  15. A simple isotopic labeling method to study cysteine oxidation in Alzheimer's disease: oxidized cysteine-selective dimethylation (OxcysDML).

    PubMed

    Gu, Liqing; Robinson, Renã A S

    2016-04-01

    Cysteine is widely involved in redox signaling pathways through a number of reversible and irreversible modifications. Reversible modifications (e.g., S-glutathionylation, S-nitrosylation, disulfide bonds, and sulfenic acid) are used to protect proteins from oxidative attack and maintain cellular homeostasis, while irreversible oxidations (e.g., sulfinic acid and sulfonic acid) serve as hallmarks of oxidative stress. Proteomic analysis of cysteine-enriched peptides coupled with reduction of oxidized thiols can be used to measure the oxidation states of cysteine, which is helpful for elucidating the role that oxidative stress plays in biology and disease. As an extension of our previously reported cysDML method, we have developed oxidized cysteine-selective dimethylation (OxcysDML), to investigate the site-specific total oxidation of cysteine residues in biologically relevant samples. OxcysDML employs (1) blocking of free thiols by a cysteine-reactive reagent, (2) enrichment of peptides containing reversibly oxidized cysteine by a solid phase resin, and (3) isotopic labeling of peptide amino groups to quantify cysteine modifications arising from different biological conditions. On-resin enrichment and labeling minimizes sample handing time and improves efficiency in comparison with other redox proteomic methods. OxcysDML is also inexpensive and flexible, as it can accommodate the exploration of various cysteine modifications. Here, we applied the method to liver tissues from a late-stage Alzheimer's disease (AD) mouse model and wild-type (WT) controls. Because we have previously characterized this proteome using the cysDML approach, we are able here to probe deeper into the redox status of cysteine in AD. OxcysDML identified 1129 cysteine sites (from 527 proteins), among which 828 cysteine sites underwent oxidative modifications. Nineteen oxidized cysteine sites had significant alteration levels in AD and represent proteins involved in metabolic processes. Overall

  16. The physics of photons and neutrons with applications of deuterium labeling methods to polymers

    SciTech Connect

    Wignall, G.D.

    1986-12-01

    Over the past decade small-angle neutron scattering (SANS), has found numerous applications in the fields of biology, polymer science, physical chemistry, materials science, metallurgy, colloids, and solid state physics. A number of excellent references are available which contain basic neutron scattering theory though these text books reflect the origins of the technique and the examples are largely drawn from physics e.g., single crystals, simple liquids, monatomic gases, liquid metals, magnetic materials, etc. in view of the large numbers of nonspecialists who are increasingly using neutron scattering, the need has become apparent for presentations which can provide rapid access to the method without unnecessary detail and mathematical rigor. This article is meant to serve as a general introduction to the symposium ''Scattering Deformation and Fracture in Polymers,'' and is intended to aid potential users who have a general scientific background, but no specialist knowledge of scattering, to apply the technique to provide new information in areas of their own particular interests. In view of space limitations, the general theory will be given in the case for neutron scattering and analogies and differences with photon scattering (x-rays) will be pointed out at the appropriate point. 90 refs., 6 figs.

  17. Radioactive labeling of proteins in cultured postimplantation mouse embryos. I. Influence of the embryo preparation method

    SciTech Connect

    Nowak, J.; Klose, J. )

    1989-07-01

    Conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations followed by fluorography. The aim was to obtain highly radioactive proteins under conditions as physiological as possible. Embryos at Days 10, 11, and 12 of gestation were prepared in different ways and incubated for 4 h in Tyrode's solution containing ({sup 3}H)amino acids (mixture) at a concentration of 27 microCi/ml medium. The preparations were: (a) yolk sac opened, placenta and blood circulation intact; (b) yolk sac and amnion opened, placenta and blood circulation intact (Day 10 embryos only); (c) placenta, yolk sac, and amnion removed (embryo naked); (d) naked embryos cut randomly into pieces (Day 10 embryos only). After incubation whole embryos or certain parts (tail, liver, rest body) were investigated by determining the radioactivity taken up by the protein. The results are given in dpm per mg protein per embryo. Radioactivity of proteins was about 3 times higher in naked embryos than in embryos left in their yolk sacs. This was true for all three stages investigated. However, the degree of radioactivity in the various parts of naked embryos differed by a factor of 15, whereas radioactivity was evenly distributed in embryos incubated in their yolk sacs. Therefore, embryos prepared according to the first method (see above) fulfilled the conditions required at the best.

  18. Radioactive labeling of proteins in cultured postimplantation mouse embryos. I. Influence of the embryo preparation method.

    PubMed

    Nowak, J; Klose, J

    1989-07-01

    Conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations followed by fluorography. The aim was to obtain highly radioactive proteins under conditions as physiological as possible. Embryos at Days 10, 11, and 12 of gestation were prepared in different ways and incubated for 4 h in Tyrode's solution containing [3H]amino acids (mixture) at a concentration of 27 microCi/ml medium. The preparations were: a) yolk sac opened, placenta and blood circulation intact; b) yolk sac and amnion opened, placenta and blood circulation intact (Day 10 embryos only); c) placenta, yolk sac, and amnion removed (embryo "naked"); d) naked embryos cut randomly into pieces (Day 10 embryos only). After incubation whole embryos or certain parts (tail, liver, rest body) were investigated by determining the radioactivity taken up by the protein. The results are given in dpm per mg protein per embryo. Radioactivity of proteins was about 3 times higher in naked embryos than in embryos left in their yolk sacs. This was true for all three stages investigated. However, the degree of radioactivity in the various parts of naked embryos differed by a factor of 15, whereas radioactivity was evenly distributed in embryos incubated in their yolk sacs. Therefore, embryos prepared according to the first method (see above) fulfilled the conditions required at the best.

  19. Automated Analysis of Human Sperm Number and Concentration (Oligospermia) Using Otsu Threshold Method and Labelling

    NASA Astrophysics Data System (ADS)

    Susrama, I. G.; Purnama, K. E.; Purnomo, M. H.

    2016-01-01

    Oligospermia is a male fertility issue defined as a low sperm concentration in the ejaculate. Normally the sperm concentration is 20-120 million/ml, while Oligospermia patients has sperm concentration less than 20 million/ml. Sperm test done in the fertility laboratory to determine oligospermia by checking fresh sperm according to WHO standards in 2010 [9]. The sperm seen in a microscope using a Neubauer improved counting chamber and manually count the number of sperm. In order to be counted automatically, this research made an automation system to analyse and count the sperm concentration called Automated Analysis of Sperm Concentration Counters (A2SC2) using Otsu threshold segmentation process and morphology. Data sperm used is the fresh sperm directly in the analysis in the laboratory from 10 people. The test results using A2SC2 method obtained an accuracy of 91%. Thus in this study, A2SC2 can be used to calculate the amount and concentration of sperm automatically

  20. METHOD OF OPERATING A HEAVY WATER MODERATED REACTOR

    DOEpatents

    Vernon, H.C.

    1962-08-14

    A method of removing fission products from the heavy water used in a slurry type nuclear reactor is described. According to the process the slurry is steam distilled with carbon tetrachloride so that at least a part of the heavy water and carbon tetrachloride are vaporized; the heavy water and carbon tetrachloride are separated; the carbon tetrachloride is returned to the steam distillation column at different points in the column to aid in depositing the slurry particles at the bottom of the column; and the heavy water portion of the condensate is purified. (AEC)

  1. Systems and Methods for Automated Water Detection Using Visible Sensors

    NASA Technical Reports Server (NTRS)

    Rankin, Arturo L. (Inventor); Matthies, Larry H. (Inventor); Bellutta, Paolo (Inventor)

    2016-01-01

    Systems and methods are disclosed that include automated machine vision that can utilize images of scenes captured by a 3D imaging system configured to image light within the visible light spectrum to detect water. One embodiment includes autonomously detecting water bodies within a scene including capturing at least one 3D image of a scene using a sensor system configured to detect visible light and to measure distance from points within the scene to the sensor system, and detecting water within the scene using a processor configured to detect regions within each of the at least one 3D images that possess at least one characteristic indicative of the presence of water.

  2. Composition and method for detecting cancer with technetium labeled antibody fragments

    SciTech Connect

    Burchiel, S. W.; Crockford, D. R.; Rhodes, B. A.

    1984-10-23

    F(ab')/sub 2/ or Fab fragments of antibodies to: (a) human chorionic gonadotropin (hCG), hCG alpha subunit, hCG beta subunit, or an hCG-like material; or (b) other tumor specific or tumor associated molecules, to include carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), human melanoma associated antigens, human sarcoma associated antigens or other antigens, are radiolabeled with technetium-99m (Tc-99m). When the F(ab')/sub 2/ or Fab fragments of antibody to such tumor associated antigens are injected intravenously into a patient, the radiolabeled composition accumulates at tumor sites. The accumulation of the cancer seeking radiopharmaceutical at tumor sites permits detection by external gamma scintigraphy. Thus, the composition is useful in the monitoring, localization and detection of cancer in the body. In an alternative composition, a double antibody approach to tumor localization using radiolabeled F(ab')/sub 2/ or Fab fragments is utilized. In this approach, a tumor specific antibody in the form of IgG, F(ab')/sub 2/ or Fab is first administered to a patient intravenously. Following a sufficient period of time, a second antibody in the form of F(ab')/sub 2/ or Fab is administered. The second antibody is radiolabeled with Tc-99m and has the property that it is reactive with the first antibody. This double antibody method has the advantage over a single antibody approach in that smaller tumors can be localized and detected and that the total amount of radioactive trace localized at the cancer site is increased.

  3. Dietary energy requirements of young adult men, determined by using the doubly labeled water method

    SciTech Connect

    Roberts, S.B.; Heyman, M.B.; Evans, W.J.; Fuss, P.; Tsay, R.; Young, V.R. )

    1991-09-01

    The autors examined the hypothesis that current recommendations on dietary energy requirements may underestimate the total energy needs of young adult men, by measuring total energy expenditure (TEE) and resting energy expenditure (REE) in 14 weight-maintaining healthy subjects leading unrestricted lives. TEE and body composition were measured by using 2H(2)18O, and REE was measured by using indirect calorimetry. All subjects had sedentary full-time occupations and participated in strenuous leisure activities for 34 {plus minus} 6 (SE) min/d. TEE and REE were 14.61 {plus minus} 0.76 and 7.39 {plus minus} 0.26 MJ/d, respectively, and 202 {plus minus} 2 and 122 {plus minus} 2 kJ.kg-1.d-1. There were significant relationships between TEE and both body fat-free mass (r = 0.732, P less than 0.005) and measured REE (r = 0.568, P less than 0.05). Measured TEE:REE values were significantly higher than the recommended energy requirement (1.98 {plus minus} 0.09, compared with 1.55 or 1.67, P less than 0.005). These results are consistent with the suggestion that the current recommended energy intake for young adult men may underestimate total energy needs.

  4. Comparison of Protein N-Homocysteinylation in Rat Plasma under Elevated Homocysteine Using a Specific Chemical Labeling Method

    PubMed Central

    Zang, Tianzhu; Pottenplackel, Ligi Paul; Handy, Diane E.; Loscalzo, Joseph; Dai, Shujia; Deth, Richard C.; Zhou, Zhaohui Sunny; Ma, Jisheng

    2017-01-01

    Elevated blood concentrations of homocysteine have been well established as a risk factor for cardiovascular diseases and neuropsychiatric diseases, yet the etiologic relationship of homocysteine to these disorders remains poorly understood. Protein N-homocysteinylation has been hypothesized as a contributing factor; however, it has not been examined globally owing to the lack of suitable detection methods. We recently developed a selective chemical method to label N-homocysteinylated proteins with a biotin-aldehyde tag followed by Western blotting analysis, which was further optimized in this study. We then investigated the variation of protein N-homocysteinylation in plasma from rats on a vitamin B12 deficient diet. Elevated “total homocysteine” concentrations were determined in rats with a vitamin B12 deficient diet. Correspondingly, overall levels of plasma protein N-homocysteinylation displayed an increased trend, and furthermore, more pronounced and statistically significant changes (e.g., 1.8-fold, p-value: 0.03) were observed for some individual protein bands. Our results suggest that, as expected, a general metabolic correlation exists between “total homocysteine” and N-homocysteinylation, although other factors are involved in homocysteine/homocysteine thiolactone metabolism, such as the transsulfuration of homocysteine by cystathionine β-synthase or the hydrolysis of homocysteine thiolactone by paraoxonase 1 (PON1), may play more significant or direct roles in determining the level of N-homocysteinylation. PMID:27617989

  5. An electron paramagnetic resonance method for measuring the affinity of a spin-labeled analog of cholesterol for phospholipids.

    PubMed

    Williams, Justin A; Wassall, Cynthia D; Kemple, Marvin D; Wassall, Stephen R

    2013-09-01

    Cholesterol (chol)-lipid interactions are thought to play an intrinsic role in determining lateral organization within cellular membranes. Steric compatibility of the rigid steroid moiety for ordered saturated chains contributes to the high affinity that holds chol and sphingomyelin together in lipid rafts whereas, conversely, poor affinity of the sterol for highly disordered polyunsaturated fatty acids (PUFAs) is hypothesized to drive the formation of PUFA-containing phospholipid domains depleted in chol. Here, we describe a novel method using electron paramagnetic resonance (EPR) to measure the relative affinity of chol for different phospholipids. We monitor the partitioning of 3β-doxyl-5α-cholestane (chlstn), a spin-labeled analog of chol, between large unilamellar vesicles (LUVs) and cyclodextrin (mβCD) through analysis of EPR spectra. Because the shape of the EPR spectrum for chlstn is sensitive to the very different tumbling rates of the two environments, the ratio of the population of chlstn in LUVs and mβCD can be determined directly from spectra. Partition coefficients (K(B)(A)) between lipids derived from our results for chlstn agree with values obtained for chol and confirm that decreased affinity for the sterol accompanies increasing acyl chain unsaturation. The virtue of this EPR method is that it provides a measure of chol binding that is quick, employs a commercially available probe and avoids the necessity for physical separation of LUVs and mβCD.

  6. Comparative evaluation of specific methods for labeling of Encephalitozoon cuniculi in paraffin wax-embedded tissue samples.

    PubMed

    Habenbacher, Bettina; Klang, Andrea; Fragner, Karin; Dinhopl, Nora; Künzel, Frank; Weissenböck, Herbert

    2012-03-01

    Detection of the microsporidian Encephalitozoon cuniculi in tissue samples is considered difficult. The aim of the current study was to determine whether immunohistochemistry (IHC) and in situ hybridization (ISH) represent reliable methods for the detection of E. cuniculi in postmortem tissue samples of rabbits. Paraffin-embedded tissue sections of brain and kidneys of 48 naturally infected pet rabbits, 10 negative controls, and the eyes of 3 further rabbits were used for all investigations. By IHC in 19 animals (37.3%), spores could be clearly detected and were all equally stained. By ISH using a digoxigenin-labeled oligonucleotide probe, only 6 animals (11.8%) proved undoubtedly positive. In these cases, many parasite-like objects revealed strong typical purple-black positive signals. However, several of the examined samples showed only partial staining of the pathogen or unclear results. Thus, in order to find an explanation for these inconsistent ISH results and to take a more detailed look at the different developmental stages of the organism, electron microscopy was applied. Empty spores, which had already discharged their polar filaments, prevailed in total number. Taken together, both techniques are rather insensitive, but under the condition that sufficient numbers of microsporidia are present, IHC can be recommended for specific identification of E. cuniculi in tissue samples. In contrast, ISH failed to detect some developmental stages of the organism, and, as such, ISH is therefore considered an inappropriate diagnostic method.

  7. Label-free optical characterization methods for detecting amine silanization-driven gold nanoparticle self-assembly.

    PubMed

    Roy, Shibsekhar; Dixit, Chandra K; Woolley, Robert; O'Kennedy, Richard; McDonagh, Colette

    2011-09-06

    Fluorescence lifetime correlation spectroscopy (FLCS) is presented as a single-step label-free detection method for probing the amine silanization-driven spontaneous 3D self-assembly of freestanding gold nanoparticles (GNPs) in solution. Unlike the conventional methods of studying self-assembly, for example, UV-vis spectroscopy and electron microscopy, FLCS utilizes the intrinsic gold fluorescence. The significance of this approach is to amalgamate the measurement of optical and hydrodynamic size properties simultaneously to achieve a more coherent description of the self-assembly pathway. GNP self-assembly has two-stage kinetics. Electrostatic interaction drives the initial amine silanization, and this is followed by siloxane bond formation between hydrolyzed ethoxy groups of GNP-attached APTES, resulting in the formation of micrometer-sized superstructures. The self-assembly has resulted in a 5-fold increase in the fluorescence lifetime (FL), and the FLCS study has shown an 8- to 10-fold increase in the diffusion coefficient using the pure diffusion model. This result is consistent with the transmission electron microscopy (TEM) observation, which shows a few hundred fold increase in the diameter due to assembly formation by the GNPs.

  8. Exploring the nitrogen ingestion of aphids--a new method using electrical penetration graph and (15)N labelling.

    PubMed

    Kuhlmann, Franziska; Opitz, Sebastian E W; Inselsbacher, Erich; Ganeteg, Ulrika; Näsholm, Torgny; Ninkovic, Velemir

    2013-01-01

    Studying plant-aphid interactions is challenging as aphid feeding is a complex process hidden in the plant tissue. Here we propose a combination of two well established methods to study nutrient acquisition by aphids focusing on the uptake of isotopically labelled nitrogen ((15)N). We combined the Electrical Penetration Graph (EPG) technique that allows detailed recording of aphid feeding behaviour and stable isotope ratio mass spectrometry (IRMS) to precisely measure the uptake of nitrogen. Bird cherry-oat aphids Rhopalosiphum padi L. (Hemiptera, Aphididae) fed for 24 h on barley plants (Hordeum vulgare L., cultivar Lina, Poaceae) that were cultivated with a (15)N enriched nutrient solution. The time aphids fed in the phloem was strongly positive correlated with their (15)N uptake. All other single behavioural phases were not correlated with (15)N enrichment in the aphids, which corroborates their classification as non-feeding EPG phases. In addition, phloem-feeding and (15)N enrichment of aphids was divided into two groups. One group spent only short time in the phloem phase and was unsuccessful in nitrogen acquisition, while the other group displayed longer phloem-feeding phases and was successful in nitrogen acquisition. This suggests that several factors such as the right feeding site, time span of feeding and individual conditions play a role for the aphids to acquire nutrients successfully. The power of this combination of methods for studying plant-aphid interactions is discussed.

  9. Exploring the Nitrogen Ingestion of Aphids — A New Method Using Electrical Penetration Graph and 15N Labelling

    PubMed Central

    Kuhlmann, Franziska; Opitz, Sebastian E. W.; Inselsbacher, Erich; Ganeteg, Ulrika; Näsholm, Torgny; Ninkovic, Velemir

    2013-01-01

    Studying plant-aphid interactions is challenging as aphid feeding is a complex process hidden in the plant tissue. Here we propose a combination of two well established methods to study nutrient acquisition by aphids focusing on the uptake of isotopically labelled nitrogen (15N). We combined the Electrical Penetration Graph (EPG) technique that allows detailed recording of aphid feeding behaviour and stable isotope ratio mass spectrometry (IRMS) to precisely measure the uptake of nitrogen. Bird cherry-oat aphids Rhopalosiphum padi L. (Hemiptera, Aphididae) fed for 24 h on barley plants (Hordeum vulgare L., cultivar Lina, Poaceae) that were cultivated with a 15N enriched nutrient solution. The time aphids fed in the phloem was strongly positive correlated with their 15N uptake. All other single behavioural phases were not correlated with 15N enrichment in the aphids, which corroborates their classification as non-feeding EPG phases. In addition, phloem-feeding and 15N enrichment of aphids was divided into two groups. One group spent only short time in the phloem phase and was unsuccessful in nitrogen acquisition, while the other group displayed longer phloem-feeding phases and was successful in nitrogen acquisition. This suggests that several factors such as the right feeding site, time span of feeding and individual conditions play a role for the aphids to acquire nutrients successfully. The power of this combination of methods for studying plant-aphid interactions is discussed. PMID:24376642

  10. A novel method to measure isotopic labeled gas-phase nitrous acid (HO15NO) in biogeochemical studies

    NASA Astrophysics Data System (ADS)

    Wu, Dianming; Kampf, Christopher; Pöschl, Ulrich; Oswald, Robert; Cui, Junfang; Ermel, Michael; Hu, Chunsheng; Trebs, Ivonne; Sörgel, Matthias

    2014-05-01

    We developed a new method (gas-phase stripping-derivatization coupled to LC-MS) to measure the 15N atom percent excess (APE) of HONO in the gas-phase. Gaseous HONO is quantitatively collected and transferred to an azo dye by the well-known Griess reaction in the Long Path Absorption Photometer (LOPAP). The reaction solutions containing the dye are collected at the outflow of the LOPAP, purified by solid-phase extraction and analyzed using high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The unlabeled azo dye (C18H19O2N5S) with a monoisotopic molecular mass of 369.41 g mol-1 can be detected as its protonated molecular ion ([M+H+], M) by HPLC-MS at a retention time of 2.8 min. Due to the natural isotope distribution M + 0, M + 1, M + 2, and M + 3 ions were considered for the calculation of the 15N APE. The optimal working range was found to be between 20 and 50% for the 15N/14N ratio. The optimum pH and solvents for extraction by SPE and potential interferences are discussed. The method has been applied for the measurement of HO15NO emissions from soil in a dynamic chamber with and without spiking 15N labeled urea. Our results confirm biogenic HONO emissions from soil as HO15NO was measured after addition of 15N urea.

  11. New methods of subcooled water recognition in dew point hygrometers

    NASA Astrophysics Data System (ADS)

    Weremczuk, Jerzy; Jachowicz, Ryszard

    2001-08-01

    Two new methods of sub-cooled water recognition in dew point hygrometers are presented in this paper. The first one- impedance method use a new semiconductor mirror in which the dew point detector, the thermometer and the heaters were integrated all together. The second one an optical method based on a multi-section optical detector is discussed in the report. Experimental results of both methods are shown. New types of dew pont hydrometers of ability to recognized sub-cooled water were proposed.

  12. Rapid method for the preparation of 125I-labelled human growth hormone for receptor studies, using reverse-phase high performance liquid chromatography

    SciTech Connect

    Ilondo, M.M.; Dehart, I.; De Meyts, P.

    1986-01-29

    Human growth hormone was labelled with 125 Iodine by the stoichiometric modification of the chloramine-T method to a specific activity of 50-80 microCi/microgram, and the iodinated mixture was purified by reverse-phase high performance liquid chromatography using a C18 column (SynChropak RP-P) and a linear gradient. Compared with the usual Sephadex G-100 chromatography, HPLC gave a much better separation, with a higher yield and a considerably reduced analysis time (30 min vs 5 h). The (125I)-labelled preparation had