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Sample records for labeled water method

  1. Validation of doubly labeled water method using a ruminant

    SciTech Connect

    Fancy, S.G.; Blanchard, J.M.; Holleman, D.F.; Kokjer, K.J.; White, R.G.

    1986-07-01

    CO/sub 2/ production (CDP, ml CO/sub 2/ . g-1 . h-1) by captive caribou and reindeer (Rangifer tarandus) was measured using the doubly labeled water method (/sup 3/H/sub 2/O and H2(18)O) and compared with CO/sub 2/ expiration rates (VCO/sub 2/), adjusted for CO/sub 2/ losses in CH4 and urine, as determined by open-circuit respirometry. CDP calculated from samples of blood or urine from a reindeer in winter was 1-3% higher than the adjusted VCO/sub 2/. Differences between values derived by the two methods of 5-20% were found in summer trials with caribou. None of these differences were statistically significant (P greater than 0.05). Differences in summer could in part be explained by the net deposition of /sup 3/H, 18O, and unlabeled CO/sub 2/ in antlers and other growing tissues. Total body water volumes calculated from /sup 3/H/sub 2/O dilution were up to 15% higher than those calculated from H/sub 2/(18)O dilution. The doubly labeled water method appears to be a reasonably accurate method for measuring CDP by caribou and reindeer in winter when growth rates are low, but the method may overestimate CDP by rapidly growing and/or fattening animals.

  2. 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.

  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. 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.

  6. Dual-Label Radioisotope Method for Simultaneously Measuring Bacterial Production and Metabolism in Natural Waters

    PubMed Central

    Jonas, Robert B.; Tuttle, Jon H.; Stoner, Daphne L.; Ducklow, Hugh W.

    1988-01-01

    Bacterial production and amino acid metabolism in aquatic systems can be estimated by simultaneous incubation of water samples with both tritiated methyl-thymidine and 14C-labeled amino acids. This dual-label method not only saves time, labor, and materials, but also allows determination of these two parameters in the same microbial subcommunity. Both organic carbon incorporation and respiration can be estimated. The results obtained with the dual-label technique are not significantly different from single-radiolabel methods over a wide range of bacterial activity. The method is particularly suitable for large-scale field programs and has been used successfully with eutrophic estuarine samples as well as with oligotrophic oceanic water. In the mesohaline portion of Chesapeake Bay, thymidine incorporation ranged seasonally from 2 to 635 pmol liter−1 h−1 and amino acid turnover rates ranged from 0.01 to 28.4% h−1. Comparison of thymidine incorporation with amino acid turnover measurements made at a deep, midbay station in 1985 suggested a close coupling between bacterial production and amino acid metabolism during most of the year. However, production-specific amino acid turnover rates increased dramatically in deep bay waters during the spring phytoplankton bloom, indicating transient decoupling of bacterial production from metabolism. Ecological features such as this are readily detectable with the dual-label method. PMID:16347587

  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. Dual-label radioisotope method for simultaneously measuring bacterial production and metabolism in natural waters

    SciTech Connect

    Jonas, B.J.; Tuttle, J.H.; Stoner, D.L.; Ducklow, H.W.

    1988-03-01

    Bacterial production and amino acid metabolism in aquatic systems can be estimated by simultaneous incubation of water samples with both tritiated methyl-thymidine and /sup 14/C-labeled amino acids. This dual-label method not only saves time, labor, and materials, but also allows determination of these two parameters in the same microbial subcommunity. Both organic carbon incorporation and respiration can be estimated. The method is particularly suitable for large-scale field programs and has been used successfully with eutrophic estuarine samples as well as with oligotrophic oceanic water. In the mesohaline portion of Chesapeake Bay, thymidine incorporation ranged seasonally from 2 to 635 pmol liter/sup -1/ h/sup -1/ and amino acid turnover rates ranged from 0.01 to 28.4% h/sup -1/. Comparison of thymidine incorporation with amino acid turnover measurements made at a deep, midbay station in 1985 suggested a close coupling between bacterial production and amino acid metabolism during most of the year. However, production-specific amino acid turnover rates increased dramatically in deep bay waters during the spring phytoplankton bloom, indicating transient decoupling of bacterial production from metabolism. Ecological features such as this are readily detectable with the dual-label method.

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

    PubMed

    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. PMID:2496076

  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. 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.

  13. Applicability of the doubly labelled water method to the rhinoceros auklet, Cerorhinca monocerata

    PubMed Central

    Shirai, Masaki; Ito, Motohiro; Yoda, Ken; Niizuma, Yasuaki

    2012-01-01

    Summary The doubly labelled water (DLW) method is an isotope-based technique that is used to measure the metabolic rates of free-living animals. We validated the DLW method for measuring metabolic rates in five rhinoceros auklets (Cerorhinca monocerata) compared with simultaneous measurements using the respirometric method. We calculated the CO2 production rate of four auklets (mean initial body mass: 552 g±36 s.d.) injected with DLW, using the one- and two-pool models. The metabolic rate during the 24-h measurements in a respirometric chamber for resting auklets averaged 16.30±1.66 kJ h−1 (n = 4). The metabolic rates determined using the one- and two-pool models in the DLW method for the same period as the respirometric measurement averaged 16.61±2.13 kJ h−1 (n = 4) and 16.16±2.10 kJ h−1 (n = 4), respectively. The mean absolute percent error between the DLW and respirometric methods was 8.04% using the one-pool model and was slightly better than that with the two-pool model. The differences in value between the DLW and respirometric methods are probably due to oxygen isotope turnover, which eliminated only 10–14% of the initial enrichment excess. PMID:23213394

  14. Methane production in ruminants: Its effect on the doubly labeled water method

    SciTech Connect

    Midwood, A.J.; Haggarty, P.; McGaw, B.A.; Robinson, J.J. )

    1989-12-01

    The doubly labeled water (DLW) technique for measuring CO2 production (rCO2) in free-living animals requires an assessment of the elimination of both 2H and 18O from the body over a long period of time. To calculate rCO2, it is necessary to calculate water turnover (rH2O) from the 2H flux rate. In ruminant animals, the accuracy of this calculation is affected by the loss of 2H in methane. We have quantified the effect of methane production (rCH4) on the 2H flux rate, determined in four sheep given 2H2O. The methane produced was depleted in 2H relative to the urine. A relationship between the enrichment of the methane and urine was established. The ratio of urine to methane enrichment was found on average to be 0.6536, and this value was unaffected by the level of rCH4 but showed some dependence on the absolute concentration of 2H in urine. For this reason, the ratio value obtained from four sheep not given 2H2O was different, a mean of 0.6886 was measured, this ratio was unaffected by changes in the diet supplied to the animals. Computer modeling was used to illustrate the dependence of the isotopically derived value for rCO2 on not only rCH4 but also the magnitude of rCO2 itself. The effect of rCH4 on the DLW method can be predicted from the observed ratio of rCO2 to rCH4 and the value of 0.6536 obtained for the ratio of methane to urine enrichment. With the use of data from several studies at this Institute, a limited range of 10 to 20 was found for rCO2/rCH4 in animals fed at or above maintenance.

  15. A Novel Method for Relative Quantitation of N-Glycans by Isotopic Labeling Using 18O-Water

    PubMed Central

    Tao, Shujuan; Orlando, Ron

    2014-01-01

    Quantitation is an essential aspect of comprehensive glycomics study. Here, a novel isotopic-labeling method is described for N-glycan quantitation using 18O-water. The incorporation of the 18O-labeling into the reducing end of N-glycans is simply and efficiently achieved during peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase F release. This process provides a 2-Da mass difference compared with the N-glycans released in 16O-water. A mathematical calculation method was also developed to determine the 18O/16O ratios from isotopic peaks. Application of this method to several standard glycoprotein mixtures and human serum demonstrated that this method can facilitate the relative quantitation of N-glycans over a linear dynamic range of two orders, with high accuracy and reproducibility. PMID:25365792

  16. Total energy expenditure of elite synchronized swimmers measured by the doubly labeled water method.

    PubMed

    Ebine, N; Feng, J Y; Homma, M; Saitoh, S; Jones, P J

    2000-09-01

    To determine the daily energy requirement of elite synchronized swimmers during moderate-intensity training, the average daily energy expenditure measured by the doubly labeled water method, was calculated for nine female Japanese national team synchronized swimmers [four senior; mean (SD) 22.5 (1.0) years old, 52.2 (3.6) kg, and five junior; 17.6 (1.1) years old, 52.8 (2.3) kg]. Their total energy expenditure (TEE) was 11.5 (2.8) MJ x day(-1) [2738 (672) kcal day(-1)]. When compared with estimated energy requirements derived from "Recommended Dietary Allowances for the Japanese", 12.1 (0.6) MJ day(-1) [2897 (139) kcal day(-1)], there was no difference between mean actual and estimated energy requirements. However, there were considerable differences observed on an individual basis. Their energy intake, estimated from 7- day self-reported dietary records, was 8.9 (1.7) MJ day(-1) [2128 (395) kcal x day(-1)], which was significantly lower than their TEE (P<0.05). Resting energy expenditure (REE), as determined by indirect calorimetry, was 5.2 (0.3) MJ x day(-1) [1247 (75) kcal x day(-1)]. Their physical activity level (TEE/REE) was 2.18 (0.43). These results demonstrate that the TEE values of elite female synchronized swimmers are not dissimilar to those reported for athletes participating in other sports, especially competitive swimmers during moderate-intensity training. PMID:11072766

  17. Methane production in ruminants: its effect on the doubly labeled water method.

    PubMed

    Midwood, A J; Haggarty, P; McGaw, B A; Robinson, J J

    1989-12-01

    The doubly labeled water (DLW) technique for measuring CO2 production (rCO2) in free-living animals requires an assessment of the elimination of both 2H and 18O from the body over a long period of time. To calculate rCO2, it is necessary to calculate water turnover (rH2O) from the 2H flux rate. In ruminant animals, the accuracy of this calculation is affected by the loss of 2H in methane. We have quantified the effect of methane production (rCH4) on the 2H flux rate, determined in four sheep given 2H2O. The methane produced was depleted in 2H relative to the urine. A relationship between the enrichment of the methane and urine was established. The ratio of urine to methane enrichment was found on average to be 0.6536, and this value was unaffected by the level of rCH4 but showed some dependence on the absolute concentration of 2H in urine. For this reason, the ratio value obtained from four sheep not given 2H2O was different, a mean of 0.6886 was measured, this ratio was unaffected by changes in the diet supplied to the animals. Computer modeling was used to illustrate the dependence of the isotopically derived value for rCO2 on not only rCH4 but also the magnitude of rCO2 itself. The effect of rCH4 on the DLW method can be predicted from the observed ratio of rCO2 to rCH4 and the value of 0.6536 obtained for the ratio of methane to urine enrichment. With the use of data from several studies at this Institute, a limited range of 10 to 20 was found for rCO2/rCH4 in animals fed at or above maintenance.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Dilution space ratio of 2H and 18O of doubly labeled water method in humans.

    PubMed

    Sagayama, Hiroyuki; Yamada, Yosuke; Racine, Natalie M; Shriver, Timothy C; Schoeller, Dale A

    2016-06-01

    Variation of the dilution space ratio (Nd/No) between deuterium ((2)H) and oxygen-18 ((18)O) impacts the calculation of total energy expenditure (TEE) by doubly labeled water (DLW). Our aim was to examine the physiological and methodological sources of variation of Nd/No in humans. We analyzed data from 2,297 humans (0.25-89 yr old). This included the variables Nd/No, total body water, TEE, body mass index (BMI), and percent body fat (%fat). To differentiate between physiologic and methodologic sources of variation, the urine samples from 54 subjects were divided and blinded and analyzed separately, and repeated DLW dosing was performed in an additional 55 participants after 6 mo. Sex, BMI, and %fat did not significantly affect Nd/No, for which the interindividual SD was 0.017. The measurement error from the duplicate urine sample sets was 0.010, and intraindividual SD of Nd/No in repeats experiments was 0.013. An additional SD of 0.008 was contributed by calibration of the DLW dose water. The variation of measured Nd/No in humans was distributed within a small range and measurement error accounted for 68% of this variation. There was no evidence that Nd/No differed with respect to sex, BMI, and age between 1 and 80 yr, and thus use of a constant value is suggested to minimize the effect of stable isotope analysis error on calculation of TEE in the DLW studies in humans. Based on a review of 103 publications, the average dilution space ratio is 1.036 for individuals between 1 and 80 yr of age. PMID:26989221

  19. 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.

  20. Energy budget in free-living animals: a novel approach based on the doubly labeled water method.

    PubMed

    Kam, M; Degen, A A

    1997-04-01

    We provide a theoretical and practical model for the calculation of energy balance of free-living animals using the doubly labeled water method. Energy expenditure, metabolizable energy intake, and body energy balance (energy retention, negative or positive) of animals are estimated using CO2 production, water influx, and dietary habits. This model accounts for CO2 produced from the 1) oxidation of dietary substrates, 2) catabolism of body tissue, and 3) deposition of body energy. We examined the model using data from studies on five homeotherms reported in the literature. The ratios between daily energy expenditure using our model and that presented in the reports ranged between 0.76 and 1.18. Metabolizable energy intakes were as low as 43% of energy expenditure in negative energy-balanced hummingbirds and as high as 245% of energy expenditure in positive energy-balanced koala bears. This model is the first that allows theoretical calculation of all energy budget components, including energy retention, in free-living animals using the doubly labeled water method. PMID:9140038

  1. 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

  2. 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.

  3. 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.

  4. 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).

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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. PMID:27573183

  12. 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

  13. 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

  14. 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.

  15. Comparison of physical activity energy expenditure in Japanese adolescents assessed by EW4800P triaxial accelerometry and the doubly labelled water method.

    PubMed

    Ishikawa-Takata, Kazuko; Kaneko, Kayoko; Koizumi, Kayo; Ito, Chinatsu

    2013-10-01

    The present study compared the accuracy of triaxial accelerometry and the doubly labelled water (DLW) method for measuring physical activity (PA) in Japanese adolescents. A total of sixty adolescents aged 12-15 years were analysed. The total energy expenditure (TEE) was measured over 7 d by the DLW method and with an EW4800P triaxial accelerometer (Panasonic Corporation). The measured (RMR)(m) and predicted RMR (RMR(p)) were 5·7 (SD 0·9) and 6·0 (SD 1·0) MJ/d, respectively. TEE measured by the DLW method and accelerometry using RMR(m) or RMR(p) were 11·0 (SD 2·6), 10·3 (SD 1·9), and 10·7 (SD 2·1) MJ/d, respectively. The PA levels (PAL) measured by the DLW method using RMR(m) or RMR(p) were 1·97 (SD 0·31) and 1·94 (SD 0·31) in subjects who exercised, and 1·85 (SD 0·27) and 1·74 (SD 0·29) in subjects who did not exercise. The percentage of body fat correlated significantly with the percentage difference between RMR(m) v. RMR(p), TEE, PA energy expenditure (PAEE) and PAL using RMR(p), and PAL using RMR(m) assessed by the DLW method and accelerometry. The present data showed that while accelerometry estimated TEE accurately, it did not provide the precise measurement of PAEE and PAL. The error in accelerometry was attributed to the prediction error of RMR and assessment in exercise. PMID:23544366

  16. Comparison of energy intake by semiquantitative food-frequency questionnaire with total energy expenditure by the doubly labeled water method in young children.

    PubMed

    Kaskoun, M C; Johnson, R K; Goran, M I

    1994-07-01

    We assessed the validity of a semiquantitative food-frequency questionnaire to estimate energy intake in young children by comparison with total energy expenditure (TEE). TEE was measured in 45 children (22 males and 23 females; 4.2-6.9 y of age) by the doubly labeled water method and body composition was estimated from bioelectrical resistance (20.2 +/- 4.0 kg body weight, 4.6 +/- 2.1 kg fat mass, and 15.6 +/- 3.1 kg fat-free mass). The sample included 36 white children and 9 Mohawk Native American children. The children's mothers completed one Willett food-frequency questionnaire to reflect the child's usual dietary intake over the last year. Total energy intake by food-frequency questionnaire (9.12 +/- 2.28 MJ/d) was significantly higher than TEE (5.74 +/- 1.13 MJ/d; P < 0.001). Misreporting of intake by food-frequency questionnaire ranged from 9.57 MJ/d overestimation to 1.58 MJ/d underestimation and was not significantly influenced by sex or body composition of the children. We conclude that use of the food-frequency questionnaire significantly overestimates energy intake in children. PMID:8017336

  17. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 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...

  18. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 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...

  19. 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...

  20. 16 CFR 301.27 - Label and method of affixing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 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...

  1. 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...

  2. Comparison of energy intake in toddlers assessed by food frequency questionnaire and total energy expenditure measured by the doubly labeled water method.

    PubMed

    Collins, Clare E; Burrows, Tracy L; Truby, Helen; Morgan, Philip J; Wright, Ian M R; Davies, Peter S W; Callister, Robin

    2013-03-01

    The ability of parents to accurately report energy intake in toddlers has rarely been validated using the gold-standard doubly labeled water (DLW) method to assess total energy expenditure (TEE). The aim of the study was to evaluate the accuracy of toddler energy intake (EI), estimated using the Australian Child and Adolescent Eating Survey (ACAES) food frequency questionnaire (FFQ) by parent report compared with a weighed food record (WFR) and TEE measured by DLW. Twelve toddlers had TEE assessed over 10 days using DLW. Usual energy intake was estimated by the primary caregiver, using standard toddler portions in ACAES-FFQ and a 4-day WFR and daily EI (in kilocalories) derived using national nutrient databases. Accuracy of reporting was calculated from absolute (EI-TEE) and percentage (EI/TEE×100) differences between EI and TEE and Pearson correlations and limits of agreement from Bland-Altman plots. Toddlers (n=12, 7 boys) had a mean age of 3.2±0.5 years, body mass index 16.2±0.9 kg, body mass index z score 0.1±0.8, EI from ACAES-FFQ 1,183±281kcal/day, and WFR 1,179±278 kcal/day and DLW TEE 1,251±149 kcal/day. The mean difference and limits of agreement (±2 standard deviations) compared with DLW was -68 (-623, 488) kcal/day for the FFQ and for the WFR -72 (-499, 354) kcal/day. Although both a semiquantitative FFQ and WFR can adequately estimate toddler energy intake at the group level in this population, toddler-specific portion size estimates should be assigned to foods listed in the FFQ. Choice of method is likely to depend on practical issues, including cost and burden. PMID:23317500

  3. 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

  4. 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.

  5. 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.

  6. Cost-benefit analysis of mollusc eating in a shorebird. I. Foraging and processing costs estimated by the doubly labelled water method.

    PubMed

    Piersma, Theunis; Dekinga, Anne; van Gils, Jan A; Achterkamp, Bart; Visser, G Henk

    2003-10-01

    Although the energy costs of foraging and food processing in vertebrates may be considerable, they have rarely been quantified separately. Here we present estimates for both cost factors based on a series of trials with a shorebird, the red knot Calidris canutus, fed natural and artificial prey types under naturalistic but fully controlled indoor aviary conditions. During eight 1-day trials we successfully manipulated the extent to which the five red knots were (1) actively probing and walking (i.e. foraging) and (2) actually ingesting prey (i.e. processing food) that was (3) either hard-shelled or not (i.e. crushing). Energy expenditures, estimated by the doubly labelled water (DLW) method, calibrated for use in this particular condition, varied between 1.5 and 4 W. A hierarchical analysis of variance indicated that the crushing of hard-shelled prey entailed no extra cost. We arrived at the following breakdown of cost components under the thermoneutral conditions of the experiment: a cost of active rest/maintenance of 1.665 W, an additional cost of foraging of 0.602 W and an additional digestive processing cost of 1.082 W. These cost levels are all well within the range of expectation and are consistent with the results of a separate outdoor aviary experiment in which the thermostatic costs needed separate estimation. On the basis of the cost and performance functions of gizzards of different mass, it was shown that under the conditions of this experiment the red knots expended the bare minimum for a balanced budget, maintaining the smallest possible gizzard. Under field conditions a larger gizzard would be required.

  7. 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

  8. 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

    2015-01-01

    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. PMID:25913933

  9. 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.

  10. Robust labeling methods for copy protection of images

    NASA Astrophysics Data System (ADS)

    Langelaar, Gerhard C.; van der Lubbe, Jan C. A.; Lagendijk, Reginald L.

    1997-01-01

    In the European project SMASH a mass multimedia storage device for home usage is being developed. The success of such a storage system depends not only on technical advances, but also on the existence of an adequate copy protection method. Copy protection for visual data requires fast and robust labeling techniques. In this paper, two new labeling techniques are proposed. The first method extends an existing spatial labeling technique. This technique divides the image into blocks and searches an optimal label- embedding level for each block instead of using a fixed embedding-level for the complete image. The embedding-level for each block is dependent on a lower quality JPEG compressed version of the labeled block. The second method removes high frequency DCT-coefficients in some areas to embed a label. A JPEG quality factor and the local image structure determine how many coefficients are discarded during the labeling process. Using both methods a perceptually invisible label of a few hundred bits was embedded in a set of true color images. The label added by the spatial method is very robust against JPEG compression. However, this method is not suitable for real-time applications. Although the second DCT-based method is slightly less resistant to JPEG compression, it is more resistant to line-shifting and cropping than the first one and is suitable for real-time labeling.

  11. 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

  12. [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.

  13. 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.

  14. A diagram retrieval method with multi-label learning

    NASA Astrophysics Data System (ADS)

    Fu, Songping; Lu, Xiaoqing; Liu, Lu; Qu, Jingwei; Tang, Zhi

    2015-01-01

    In recent years, the retrieval of plane geometry figures (PGFs) has attracted increasing attention in the fields of mathematics education and computer science. However, the high cost of matching complex PGF features leads to the low efficiency of most retrieval systems. This paper proposes an indirect classification method based on multi-label learning, which improves retrieval efficiency by reducing the scope of compare operation from the whole database to small candidate groups. Label correlations among PGFs are taken into account for the multi-label classification task. The primitive feature selection for multi-label learning and the feature description of visual geometric elements are conducted individually to match similar PGFs. The experiment results show the competitive performance of the proposed method compared with existing PGF retrieval methods in terms of both time consumption and retrieval quality.

  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. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. Protease- and Acid-catalyzed Labeling Workflows Employing 18O-enriched Water

    PubMed Central

    Klingler, Diana; Hardt, Markus

    2013-01-01

    Stable isotopes are essential tools in biological mass spectrometry. Historically, 18O-stable isotopes have been extensively used to study the catalytic mechanisms of proteolytic enzymes1-3. With the advent of mass spectrometry-based proteomics, the enzymatically-catalyzed incorporation of 18O-atoms from stable isotopically enriched water has become a popular method to quantitatively compare protein expression levels (reviewed by Fenselau and Yao4, Miyagi and Rao5 and Ye et al.6). 18O-labeling constitutes a simple and low-cost alternative to chemical (e.g. iTRAQ, ICAT) and metabolic (e.g. SILAC) labeling techniques7. Depending on the protease utilized, 18O-labeling can result in the incorporation of up to two 18O-atoms in the C-terminal carboxyl group of the cleavage product3. The labeling reaction can be subdivided into two independent processes, the peptide bond cleavage and the carboxyl oxygen exchange reaction8. In our PALeO (protease-assisted labeling employing 18O-enriched water) adaptation of enzymatic 18O-labeling, we utilized 50% 18O-enriched water to yield distinctive isotope signatures. In combination with high-resolution matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS/MS), the characteristic isotope envelopes can be used to identify cleavage products with a high level of specificity. We previously have used the PALeO-methodology to detect and characterize endogenous proteases9 and monitor proteolytic reactions10-11. Since PALeO encodes the very essence of the proteolytic cleavage reaction, the experimental setup is simple and biochemical enrichment steps of cleavage products can be circumvented. The PALeO-method can easily be extended to (i) time course experiments that monitor the dynamics of proteolytic cleavage reactions and (ii) the analysis of proteolysis in complex biological samples that represent physiological conditions. PALeO-TimeCourse experiments help identifying rate-limiting processing

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... method of affixing. (a) A label is required to be affixed to each wool product and, where required, to... until sold and delivered to the ultimate consumer. (b) Each wool product with a neck must have a label... label or labels on the inside or outside of the product. The country of origin disclosure must...

  9. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... method of affixing. (a) A label is required to be affixed to each wool product and, where required, to... until sold and delivered to the ultimate consumer. (b) Each wool product with a neck must have a label... label or labels on the inside or outside of the product. The country of origin disclosure must...

  10. 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.

  11. Labeling DNA for single-molecule experiments: methods of labeling internal specific sequences on double-stranded DNA

    NASA Astrophysics Data System (ADS)

    Zohar, Hagar; Muller, Susan J.

    2011-08-01

    This review is a practical guide for experimentalists interested in specifically labeling internal sequences on double-stranded (ds) DNA molecules for single-molecule experiments. We describe six labeling approaches demonstrated in a single-molecule context and discuss the merits and drawbacks of each approach with particular attention to the amount of specialized training and reagents required. By evaluating each approach according to criteria relevant to single-molecule experiments, including labeling yield and compatibility with cofactors such as Mg2+, we provide a simple reference for selecting a labeling method for given experimental constraints. Intended for non-specialists seeking accessible solutions to DNA labeling challenges, the approaches outlined emphasize simplicity, robustness, suitability for use by non-biologists, and utility in diverse single-molecule experiments.

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

    DOEpatents

    Anderson, Peter J.; Dueker, Stephen; Miller, Joshua; Green, Ralph; Roth, John; Carkeet, Colleen; Buchholz,; Bruce A.

    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.

  13. 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.

  14. 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.

  15. A (18)F-labeled glucose analog: synthesis using a click labeling method and in vitro evaluation.

    PubMed

    Kim, Dong Hyun; Choe, Yearn Seong; Jung, Kyung-Ho; Lee, Kyung-Han; Choi, Joon Young; Choi, Yong; Kim, Byung-Tae

    2008-05-01

    A (18)F-labeled glucose analog, 4-[(2-[(18)F]fluoroethyl)-1-(beta-D: -glucopyranosyl)]-1H-1,2,3-triazole ([(18)F]1), was synthesized using a click labeling method and evaluated in vitro for its cellular transportation via glucose transporter (Glut-1) and its potential as a hexokinase substrate. The click labeling method was superior to conventional labeling method, due to a higher decay-corrected radiochemical yield (30% vs. 21%), higher specific activity (59.9 GBq/mumol vs. 23.5 GBq/mumol), and shorter synthesis time (75-80 min vs. 95-100 min). In vitro evaluation demonstrated that [(18)F]1 does not act as a hexokinase substrate and has low and non-specific uptake by SNU-C5 cells. These results suggest that click chemistry offers a rapid and efficient radiolabeling method which does not require the protection of functional groups, although a triazole moiety at C1 of [(18)F]1 is incompatible for hexokinase phosphorylation and facilitative diffusion via Glut-1. PMID:18481013

  16. 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

  17. 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.

  18. A novel method for selectively labelling olivocochlear collaterals in the rat.

    PubMed

    Baashar, Ahmaed; Robertson, Donald; Mulders, Wilhelmina H A M

    2015-07-01

    Axons of olivocochlear neurons originate from the brainstem and project to the cochlea. A subpopulation, medial olivocochlear (MOC) neurons, also projects collateral branches to the cochlear nucleus. The precise targets of these collaterals are as yet unknown. Previous methods for labelling these collaterals include firstly, cochlear injections of retrograde tracers, but this is technically demanding and can also label afferent projections or secondly, labelling by injecting tracers into the nuclei of origin of MOC neurons. However, this latter method is non-specific because it also labels non-MOC projections. A technique was used to specifically label MOC collaterals, which involved injections of the tracer biocytin at the floor of the fourth ventricle and fixation 3 hours later. Biocytin injections resulted in labelled neurons in the ventral nucleus of the trapezoid body and rostral periolivary nucleus, confirming MOC axonal labelling. Labelled neurons in dorsal cochlear nucleus indicated labelling of the dorsal acoustic stria and these injections were discarded. After selective MOC labelling, collateral branches were found to innervate granule cell regions, medial edge and core of the ventral cochlear nucleus, as well as the dorsal cochlear nucleus, in agreement with previous data. Therefore we conclude that injections at the floor of the fourth ventricle provide a simple, rapid and specific technique for labelling the majority of MOC axons and their collaterals and this technique may assist in defining the precise neuronal targets of olivocochlear collaterals in cochlear nucleus.

  19. A novel method for selectively labelling olivocochlear collaterals in the rat.

    PubMed

    Baashar, Ahmaed; Robertson, Donald; Mulders, Wilhelmina H A M

    2015-07-01

    Axons of olivocochlear neurons originate from the brainstem and project to the cochlea. A subpopulation, medial olivocochlear (MOC) neurons, also projects collateral branches to the cochlear nucleus. The precise targets of these collaterals are as yet unknown. Previous methods for labelling these collaterals include firstly, cochlear injections of retrograde tracers, but this is technically demanding and can also label afferent projections or secondly, labelling by injecting tracers into the nuclei of origin of MOC neurons. However, this latter method is non-specific because it also labels non-MOC projections. A technique was used to specifically label MOC collaterals, which involved injections of the tracer biocytin at the floor of the fourth ventricle and fixation 3 hours later. Biocytin injections resulted in labelled neurons in the ventral nucleus of the trapezoid body and rostral periolivary nucleus, confirming MOC axonal labelling. Labelled neurons in dorsal cochlear nucleus indicated labelling of the dorsal acoustic stria and these injections were discarded. After selective MOC labelling, collateral branches were found to innervate granule cell regions, medial edge and core of the ventral cochlear nucleus, as well as the dorsal cochlear nucleus, in agreement with previous data. Therefore we conclude that injections at the floor of the fourth ventricle provide a simple, rapid and specific technique for labelling the majority of MOC axons and their collaterals and this technique may assist in defining the precise neuronal targets of olivocochlear collaterals in cochlear nucleus. PMID:25814172

  20. A novel method for observing proteins in vivo using a small fluorescent label and multiphoton imaging.

    PubMed

    Botchway, Stanley W; Barba, Ignasi; Jordan, Randolf; Harmston, Rebecca; Haggie, Peter M; Williams, Simon-Peter; Fulton, Alexandra M; Parker, Anthony W; Brindle, Kevin M

    2005-09-15

    A novel method for the fluorescence detection of proteins in cells is described in the present study. Proteins are labelled by the selective biosynthetic incorporation of 5-hydroxytryptophan and the label is detected via selective two-photon excitation of the hydroxyindole and detection of its fluorescence emission at 340 nm. The method is demonstrated in this paper with images of a labelled protein in yeast cells.

  1. A novel dual-isotope labelling method for distinguishing between soil sources of N2O.

    PubMed

    Wrage, N; van Groenigen, J W; Oenema, O; Baggs, E M

    2005-01-01

    We present a novel 18O-15N-enrichment method for the distinction between nitrous oxide (N2O) from nitrification, nitrifier denitrification and denitrification based on a method with single- and double-15N-labelled ammonium nitrate. We added a new treatment with 18O-labelled water to quantify N2O from nitrifier denitrification. The theory behind this is that ammonia oxidisers use oxygen (O2) from soil air for the oxidation of ammonia (NH3), but use H2O for the oxidation of the resulting hydroxylamine (NH2OH) to nitrite (NO2-). Thus, N2O from nitrification would therefore be expected to reflect the 18O signature of soil O2, whereas the 18O signature of N2O from nitrifier denitrification would reflect that of both soil O2 and H2O. It was assumed that (a) there would be no preferential removal of 18O or 16O during nitrifier denitrification or denitrification, (b) the 18O signature of the applied 18O-labelled water would remain constant over the experimental period, and (c) any O exchange between H(2)18O and NO3- would be negligible under the chosen experimental conditions. These assumptions were tested and validated for a silt loam soil at 50% water-filled pore space (WFPS) following application of 400 mg N kg-1 dry soil. We compared the results of our new method with those of a conventional inhibition method using 0.02% v/v acetylene (C2H2) and 80% v/v O2 in helium. Both the 18O-15N-enrichment and inhibitor methods identified nitrifier denitrification to be a major source of N2O, accounting for 44 and 40%, respectively, of N2O production over 24 h. However, compared to our 18O-15N-method, the inhibitor method overestimated the contribution from nitrification at the expense of denitrification, probably due to incomplete inhibition of nitrifier denitrification and denitrification by large concentrations of O2 and a negative effect of C2H2 on denitrification. We consider our new 18O-15N-enrichment method to be more reliable than the use of inhibitors; it enables the

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

    DOEpatents

    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.

  3. Nanoparticles labeled with Positron Emitting Nuclides: Advantages, Methods, and Applications

    PubMed Central

    Liu, Yongjian; Welch, Michael J.

    2012-01-01

    Over the past decade, positron emitter labeled nanoparticles have been widely used in and substantially improved for a range of diagnostic biomedical research. However, given growing interest in personalized medicine and translational research, a major challenge in the field will be to develop disease specific nanoprobes with facile and robust radiolabeling strategies and that provide imaging stability, enhanced sensitivity for disease early stage detection, optimized in vivo pharmacokinetics for reduced non-specific organ uptake, and improved targeting for elevated efficacy. This review briefly summarizes the major applications of nanoparticles labeled with positron emitters for cardiovascular imaging, lung diagnosis and tumor theranostics. PMID:22242601

  4. 40 CFR 600.115-11 - Criteria for determining the fuel economy label calculation method.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 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...

  5. Energy expenditure by doubly labeled water: validation in humans and proposed calculation

    SciTech Connect

    Schoeller, D.A.; Ravussin, E.; Schutz, Y.; Acheson, K.J.; Baertschi, P.; Jequier, E.

    1986-05-01

    To further validate the doubly labeled water method for measurement of CO/sub 2/ production and energy expenditure in humans, we compared it with near-continuous respiratory gas exchange in nine healthy young adult males. Subjects were housed in a respiratory chamber for 4 days. Each received /sup 2/H/sub 2/(18)O at either a low (n = 6) or a moderate (n = 3) isotope dose. Low and moderate doses produced initial /sup 2/H enrichments of 5 and 10 X 10(-3) atom percent excess, respectively, and initial 18O enrichments of 2 and 2.5 X 10(-2) atom percent excess, respectively. Total body water was calculated from isotope dilution in saliva collected at 4 and 5 h after the dose. CO/sub 2/ production was calculated by the two-point method using the isotopic enrichments of urines collected just before each subject entered and left the chamber. Isotope enrichments relative to predose samples were measured by isotope ratio mass spectrometry. At low isotope dose, doubly labeled water overestimated average daily energy expenditure by 8 +/- 9% (SD) (range -7 to 22%). At moderate dose the difference was reduced to +4 +/- 5% (range 0-9%). The isotope elimination curves for /sup 2/H and 18O from serial urines collected from one of the subjects showed expected diurnal variations but were otherwise quite smooth. The overestimate may be due to approximations in the corrections for isotope fractionation and isotope dilution. An alternative approach to the corrections is presented that reduces the overestimate to 1%.

  6. Preparation of 99mTc-TRODAT-1 with high labeling yield in boiling water bath: a new formulation.

    PubMed

    Erfani, Mostafa; Shafiei, Mohammad

    2014-04-01

    A new formulation for preparation of (99m)Tc-labeled tropane derivative, (99m)Tc-TRODAT-1, which is useful as a potential CNS dopamine transporter imaging agent, was evaluated and characterized. Preparation of (99m)Tc-TRODAT-1 was attained previously by a formulation in which vial has to be autoclaved at 121 °C for 30 min. It is highly desirable to further improve the preparation method by developing a simplified one vial formulation which will be labeled in boiling water bath (95 °C) for 15 min and a high labeling yield will be achieved. A formulation contained 10 μg of TRODAT-1, 20 μg tricine, 40 μg SnCl2 and 20mg manitol was prepared. Labeling was performed at 95 °C for 15 min and radiochemical analysis involved ITLC and HPLC methods. The stability of radioconjugate was checked in the presence of human serum at 37 °C up to 24h. (99m)Tc-TRODAT-1 was prepared with a radiochemical purity of more than 95% and specific activity of 64.3 MBq/nmol. Biodistribution studies of this new formulation in rats revealed similar regional brain distribution as compared with those obtained with the previous preparation in which brain uptake was high in striatum and striatum to cerebellum ratio was high. Requiring no autoclave facility for labeling, this new formulation will significantly improve the using feasibility of this radiopharmaceutical in clinic.

  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. 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.

  9. 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.

  10. 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.

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

    PubMed

    Härmä, Harri; Peltomaa, Riikka; Pihlasalo, Sari

    2015-07-01

    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. PMID:26102618

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

    PubMed

    Härmä, Harri; Peltomaa, Riikka; Pihlasalo, Sari

    2015-07-01

    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.

  13. 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.

  14. 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.

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

    PubMed

    Trott, N G; Akbari, R B

    1984-02-01

    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. PMID:6719926

  16. Energy expenditure and fluid production in hyperbaric He-O2 environments using doubly labeled water.

    PubMed

    Seale, J L; Thorp, J W; Conway, J M; Rumpler, W V; Haberman, K J

    1994-06-01

    Energy expenditure (EE), carbon dioxide production (rCO2), water turnover (rH2O), and urine production (UP) were measured to determine nutrient requirements of U.S. Navy divers during saturation dives. Parameters were measured in a normal surface environment (n = 10) and in 0.56 MPa (n = 9) and 3.17 MPa (n = 11) helium-oxygen environments. Daily EE, rCO2, and rH2O were measured with the doubly labeled water method for 10-14 days in each environment. Daily UP was determined by 24-h urine collection for 5- to 10-day periods in each environment. Divers consumed a mixed diet composed of 30% calories from fat, 15% protein, and 55% carbohydrate. Both EE and rCO2 increased significantly relative to surface conditions at 0.56 MPa (13 +/- 4% and 11 +/- 4%) and 3.17 MPa (14 +/- 4% and 11 +/- 3%), but there was no difference between dives. Water turnover was not significantly affected by the hyperbaric environment. UP was significantly greater than surface conditions at 0.56 MPa (53 +/- 19%) but not at 3.17 MPa (38 +/- 18%). Increased EE is attributed to thermal stress caused by the helium-oxygen environment. Increased UP may have been caused by decreased evaporative water loss. PMID:7914783

  17. Fluorescent labelling of ciprofloxacin and norfloxacin and its application for residues analysis in surface water.

    PubMed

    Prutthiwanasan, Brompoj; Phechkrajang, Chutima; Suntornsuk, Leena

    2016-10-01

    Sensitivity enhancement for residue analysis of ciprofloxacin and norfloxacin in surface water was performed by liquid chromatography with fluorescent detection (LC-FD). Labelling of both drugs were studied with fluorescent probes (e.g. Nile blue perchlorate (NBP) and 4- (N,N-Dimethylaminosulfonyl)-7-(N-chloroformylmethyl-N-methylamino)-2,1,3-benzoxadiazole (DBD-COCl). Factors affecting the derivatization (e.g. stoichiometric ratios, reaction time and base catalysts) were optimized. The derivatization was achieved in 15min using a stoichiometric ratio between the substrate and DBD-COCl of 1:3, whereas NBP gave unsatisfactory results. Separation of the derivatives by LC was achieved (resolution (RS) > 1.8) on a C8 column using a mobile phase consisting of 50mM formic acid and acetonitrile (ACN) (68:32% v/v) in 20min. The method was linear (r(2) > 0.99) in a range of 200-2,000µg/L, precise (%RSD < 9.17) and accurate (%recovery of 102.5-122.2%) for the determination of the derivatives. The uses of fluoroquinolone molecularly imprinted polymer in conjunction with hydrophilic-lipophilic balance sorbents demonstrated an efficient procedure for sample pre-concentration and clean-up for water sample resulting in the improved percent recovery. Applications of the proposed method was shown in surface water samples in Thailand. PMID:27474281

  18. 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...

  19. Label-free detection repeatability of protein microarrays by oblique-incidence reflectivity difference method

    NASA Astrophysics Data System (ADS)

    Dai, Jun; Li, Lin; Wang, JingYi; He, LiPing; Lu, HuiBin; Ruan, KangCheng; Jin, KuiJuan; Yang, GuoZhen

    2012-12-01

    We examine the repeatabilities of oblique-incidence reflectivity difference (OIRD) method for label-free detecting biological molecular interaction using protein microarrays. The experimental results show that the repeatabilities are the same in a given microarray or microarray-microarray and are consistent, indicating that OIRD is a promising label-free detection technique for biological microarrays.

  20. 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. PMID:20967672

  1. Ultrasensitive one-step rapid visual detection of bisphenol A in water samples by label-free aptasensor.

    PubMed

    Mei, Zhanlong; Chu, Huaqin; Chen, Wei; Xue, Feng; Liu, Jian; Xu, Huaneng; Zhang, Rui; Zheng, Lei

    2013-01-15

    A simple, one-step, rapid method to detect bisphenol A (BPA) using a label-free aptasensor is presented. A high selective anti-BPA aptamer was added to gold nanoparticles (GNPs) to prepare the label-free aptasensor for BPA, which maintains good tolerance of GNPs under aqueous conditions with high salt concentrations. With the presence of BPA in the aptasensor system, the GNPs would aggregate by competitive binding of BPA and aptamer. Detection results can be visualized by the aggregation-induced color change of GNPs without the use of any instrumentation. The limit of visual detection (LOD) was found to be 0.1ng/mL by naked-eye observation, which was competitive to some current rapid BPA detection methods, even some instrumental based methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for BPA and are comparable to the traditional ELISA and instrument-based methods. The proposed one-step, label-free method was successfully used to determine BPA in actual water samples. PMID:22794930

  2. 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.

  3. 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. PMID:11006607

  4. 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.

  5. Using Doubly-Labeled Water to Measure Energy Expenditure in an Important Small Ectotherm Drosophila melanogaster

    PubMed Central

    Piper, Matthew D.W.; Selman, Colin; Speakman, John R.; Partridge, Linda

    2014-01-01

    Energy expenditure is a key variable in the study of ageing, and the fruit fly Drosophila melanogaster is a model organism that has been used to make step changes in our understanding of the ageing process. Standard methods for measurement of energy expenditure involve placing individuals in metabolic chambers where their oxygen consumption and CO2 production can be quantified. These measurements require separating individuals from any social context, and may only poorly reflect the environment in which the animals normally live. The doubly-labeled water (DLW) method is an isotope-based technique for measuring energy expenditure which overcomes these problems. However, technical challenges mean that the smallest animals this method has been previously applied to weighed 50–200 mg. We overcame these technical challenges to measure energy demands in Drosophila weighing 0.78 mg. Mass-specific energy expenditure varied between 43 and 65 mW·g−1. These estimates are considerably higher than estimates using indirect calorimetry of Drosophila in small metabolic chambers (around 18 mW·g−1). The methodology we have established extends downwards by three orders of magnitude the size of animals that can be measured using DLW. This approach may be of considerable value in future ageing research attempting to understand the genetic and genomic basis of ageing. PMID:25269676

  6. Using doubly-labeled water to measure energy expenditure in an important small ectotherm Drosophila melanogaster.

    PubMed

    Piper, Matthew D W; Selman, Colin; Speakman, John R; Partridge, Linda

    2014-09-20

    Energy expenditure is a key variable in the study of ageing, and the fruit fly Drosophila melanogaster is a model organism that has been used to make step changes in our understanding of the ageing process. Standard methods for measurement of energy expenditure involve placing individuals in metabolic chambers where their oxygen consumption and CO2 production can be quantified. These measurements require separating individuals from any social context, and may only poorly reflect the environment in which the animals normally live. The doubly-labeled water (DLW) method is an isotope-based technique for measuring energy expenditure which overcomes these problems. However, technical challenges mean that the smallest animals this method has been previously applied to weighed 50-200 mg. We overcame these technical challenges to measure energy demands in Drosophila weighing 0.78 mg. Mass-specific energy expenditure varied between 43 and 65 mW·g(-1). These estimates are considerably higher than estimates using indirect calorimetry of Drosophila in small metabolic chambers (around 18 mW·g(-1)). The methodology we have established extends downwards by three orders of magnitude the size of animals that can be measured using DLW. This approach may be of considerable value in future ageing research attempting to understand the genetic and genomic basis of ageing.

  7. 78 FR 43974 - Energy and Water Use Labeling for Consumer Products Under the Energy Policy and Conservation Act...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-23

    ... new range for instantaneous electric water heaters based on data submitted by industry. \\10\\ 77 FR... CFR Part 305 Energy and Water Use Labeling for Consumer Products Under the Energy Policy and Conservation Act (Energy Labeling Rule) AGENCY: Federal Trade Commission (``FTC'' or ``Commission'')....

  8. Total energy expenditure assessment in elite junior basketball players: a validation study using doubly labeled water.

    PubMed

    Silva, Analiza M; Santos, Diana A; Matias, Catarina N; Minderico, Cláudia S; Schoeller, Dale A; Sardinha, Luís B

    2013-07-01

    An accurate assessment of total energy expenditure (TEE) during a competitive season is required. We aimed to validate TEE estimated by self-reported energy intake (EI) and the dietary reference intake (DRI) method in 19 elite basketball players (aged 16-18 years) using doubly labeled water (DLW) as the reference method. The DRI models and EI from dietary records over a 7-day period were simultaneously assessed for TEE estimation. Resting energy expenditure was assessed by indirect calorimetry. Fat and fat-free mass (FFM) were determined by a 4-compartment model (body volume by air displacement plethysmography, bone mineral by DXA, and water by deuterium dilution). Fat and FFM ranged from 4 to 19 kg and from 47 to 81 kg, respectively. The physical activity level ranged from 2.2 to 3.7 with a mean value of 2.8 ± 0.4. Total energy expenditure from DLW (17,598 ± 3,298 kJ·d) was significantly underestimated by EI (11,274 ± 2,567 kJ·d), whereas no differences were found using DRI (17,008 ± 3,206 kJ·d). The EI and DRI methods explained TEE from DLW by 34% (p = 0.057) and 44% (p = 0.002), respectively, and wide limits of agreement were observed. Our findings suggested that EI is not a valid tool for TEE assessment. The DRI method may be valid at a group level but inaccurate for estimating individual TEE in young players during a demanding competitive season period.

  9. Robust MS quantification method for phospho-peptides using 18O/16O labeling

    PubMed Central

    Andersen, Claus A; Gotta, Stefano; Magnoni, Letizia; Raggiaschi, Roberto; Kremer, Andreas; Terstappen, Georg C

    2009-01-01

    Background Quantitative measurements of specific protein phosphorylation sites, as presented here, can be used to investigate signal transduction pathways, which is an important aspect of cell dynamics. The presented method quantitatively compares peptide abundances from experiments using 18O/16O labeling starting from elaborated MS spectra. It was originally developed to study signaling cascades activated by amyloid-β treatment of neurons used as a cellular model system with relevance to Alzheimer's disease, but is generally applicable. Results The presented method assesses, in complete cell lysates, the degree of phosphorylation of specific peptide residues from MS spectra using 18O/16O labeling. The abundance of each observed phospho-peptide from two cell states was estimated from three overlapping isotope contours. The influence of peptide-specific labeling efficiency was removed by performing a label swapped experiment and assuming that the labeling efficiency was unchanged upon label swapping. Different degrees of phosphorylation were reported using the fold change measure which was extended with a confidence interval found to reflect the quality of the underlying spectra. Furthermore a new way of method assessment using simulated data is presented. Using simulated data generated in a manner mimicking real data it was possible to show the method's robustness both with increasing noise levels and with decreasing labeling efficiency. Conclusion The fold change error assessable on simulated data was on average 0.16 (median 0.10) with an error-to-signal ratio and labeling efficiency distributions similar to the ones found in the experimentally observed spectra. Applied to experimentally observed spectra a very good match was found to the model (<10% error for 85% of spectra) with a high degree of robustness, as assessed by data removal. This new method can thus be used for quantitative signal cascade analysis of total cell extracts in a high throughput mode

  10. 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

  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. Improved 2-nitrobenzenesulfenyl method: optimization of the protocol and improved enrichment for labeled peptides.

    PubMed

    Matsuo, Ei-Ichi; Toda, Chikako; Watanabe, Makoto; Iida, Tetsuo; Masuda, Taro; Minohata, Toshikazu; Ando, Eiji; Tsunasawa, Susumu; Nishimura, Osamu

    2006-01-01

    We have developed the NBS (2-nitrobenzenesulfenyl) method, a quantitative proteome analysis method utilizing stable isotope labeling followed by mass spectrometry. The potential of this method was reported previously, and the procedure has now been further optimized. Here, we describe a procedure utilizing urea or guanidine hydrochloride as a protein denaturant, in conjunction with an improved chromatographic enrichment method for the NBS-labeled peptides using a phenyl resin column. By using this new protocol, both sample loss throughout the protocol and the elution of unwanted unlabeled peptides can be minimized, improving the efficiency of the analysis significantly.

  14. A new method to specifically label thiophosphorylatable proteins with extrinsic probes. Labeling of serine-19 of the regulatory light chain of smooth muscle myosin.

    PubMed

    Facemyer, K C; Cremo, C R

    1992-01-01

    We present a new method to specifically and stably label proteins by attaching extrinsic probes to amino acids that are thiophosphorylated by protein kinases and ATP gamma S. The method was demonstrated for labeling of a thiophosphorylatable serine of the isolated regulatory light chain of smooth muscle myosin. We stoichiometrically blocked the single thiol (Cys-108) either by forming a reversible intermolecular disulfide bond or by reacting with iodoacetic acid. The protein was stoichiometrically thiophosphorylated at Ser-19 by myosin light chain kinase and ATP gamma S. The nucleophilic sulfur of the protein phosphorothioate was coupled at pH 7.9 and 25 degrees C to the fluorescent haloacetate [3H]-5-[[2-[(iodoacetyl)-amino]ethyl]amino]naphthalene-1- sulfonic acid ([3H]IAEDANS) by displacement of the iodide. Typical labeling efficiencies were 70-100%. The labeling was specific for the thiophosphorylated Ser-19, as determined from the sequences of two labeled peptides isolated from a tryptic digest of the labeled protein. [3H]IAEDANS attached to the thiophosphorylated Ser-19 was stable at pH 3-10 at 25 degrees C, and to boiling in high concentrations of reductant. The labeled light chains were efficiently exchanged for unlabeled regulatory light chains of the whole myosin molecule. The resulting labeled myosin had normal ATPase activities in the absence of actin, indicating that the modification of Ser-19 and the exchange of the labeled light chain into myosin did not significantly disrupt the protein. The labeled myosin partially retained the elevated actin-activated Mg(2+)-ATPase activity which is characteristic of thiophosphorylated myosin. This indicates that labeling of the thiophosphate group with [3H]IAEDANS did not completely disrupt the functional properties of the thiophosphorylated protein in the presence of actin.

  15. 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

  16. 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

  17. Improvement of the periodate-borohydride surface-labeling method for human blood platelets

    SciTech Connect

    Steiner, B.; Clemetson, K.J.; Luescher, E.F.

    1983-01-01

    The periodate/sodium boro(/sup 3/H)hydride ((/sup 3/H)-NaBH4) method is extensively used for the specific labeling of cell surface glycoproteins. Reduction with tritiated borohydride is also used in other surface-labeling techniques, the neuraminidase/galactose oxidase/(/sup 3/H)-NaBH4 method (specific for terminal galactose and N-acetyl-galactosamine residues) and the pyridoxal phosphate/(/sup 3/H)-NaBH4 method (specific for protein). By modification of the reaction conditions during the periodate-oxidation and borohydride-reduction, the ratio of the incorporated to the total added radioactivity could be increased by a factor of 50, while the specific activity of the labeled material was twice as high as in the original method. Alternatively, by another modification, the specific activity of the labeled material could be increased about 10-fold. The influence of the most important parameters was investigated in detail. Sodium dodecyl sulfate gel electrophoresis and fluorography demonstrate that the labeling pattern of the membrane glycoproteins is the same as with the conventional method.

  18. Method for treating waste water

    SciTech Connect

    Helke, R.C.

    1980-12-02

    A method useful for treating waste water is disclosed wherein waste water is collected in a first vessel and a portion of the large solid particles are filtered from said waste waters. The liquid waste including suspended solid particles is combined with a solids coagulant, effective in coagulating solid particles, and the waste is disinfected. In one embodiment, coagulation and disinfection occurs simultaneously in a single treatment vessel. In the treatment vessel, the disinfectant and the coagulant are reacted with the waste waters to form gas bubbles and coagulated solid particles. The reaction of the disinfectant causes a substantial portion of the coagulated solids contained in the treatment vessel to float to the upper portion of the treatment vessel as a result of being carried to the surface by the gas bubbles. The clarified waste water is then removed from an outer chamber in the treatment vessel. In another embodiment, waste water is disinfected by radiation so that gas bubbles are not formed by the disinfection reaction. In this embodiment the coagulated solids are floated to the surface of the treatment vessel by providing small gas, i.e., air, bubbles in the treatment vessel generated from an extraneous source.

  19. 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

  20. Use of the doubly labeled water technique in humans during heavy sustained exercise

    SciTech Connect

    Westerterp, K.R.; Saris, W.H.; van Es, M.; ten Hoor, F.

    1986-12-01

    We measured energy expenditure with the doubly labeled water technique during heavy sustained exercise in the Tour de France, a bicycle race lasting more than 3 wk. Four subjects were observed for consecutive intervals of 7, 8, and 7 days. Each interval started with an oral isotope dose to reach an excess isotope level of 200 ppm 18O and 130 ppm 2H. The biological half-lives of the isotopes were between 2.25 and 3.80 days. Energy expenditure was compared with simultaneous measurements of energy intake, and body mass and body composition did not change significantly. The doubly labeled water technique gave higher values for energy expenditure than the food record technique. The discrepancy showed a systematic increment from the first to the third interval, being 12.9 +/- 7.9, 21.4 +/- 9.8, and 35.3 +/- 4.4% of the energy expenditure calculated from dietary intake, respectively. Possible explanations for the discrepancy are discussed. The subjects reached an average daily metabolic rate of 3.4-3.9 or 4.3-5.3 times basal metabolic rate based on the food record technique and the doubly labeled water technique, respectively. Thus, when measured with the same technique, the energetic ceiling for performance in humans is comparable with that of animals like birds.

  1. 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

  2. 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.

  3. Evaluation of red blood cell labelling methods based on a statistical model for red blood cell survival.

    PubMed

    Korell, Julia; Coulter, Carolyn V; Duffull, Stephen B

    2011-12-21

    The aim of this work is to compare different labelling methods that are commonly used to estimate the lifespan of red blood cells (RBCs), e.g. in anaemia of renal failure, where the effect of treatment with erythropoietin depends on the lifespan of RBCs. A previously developed model for the survival time of RBCs that accounts for plausible physiological processes of RBC destruction was used to simulate ideal random and cohort labelling methods for RBCs, as well as the flaws associated with these methods (e.g. reuse of label and loss of the label from the surviving RBCs). Random labelling with radioactive chromium and cohort labelling using heavy nitrogen were considered. Blood sampling times were determined for RBC survival studies using both labelling methods by applying the theory of optimal design. It was assessed whether the underlying parameter values of the model are estimable from these studies, and the precision of the parameter estimates were calculated. In theory, parameter estimation would be possible for both types of ideal labelling methods without flaws. However, flaws associated with random labelling are significant and not all parameters controlling RBC survival in the model can be estimated with good precision. In contrast, cohort labelling shows good precision in the parameter estimates even in the presence of reuse and prolonged incorporation of the label. A model based analysis of RBC survival studies is recommended in future to account for limitations in methodology as well as likely causes of RBC destruction. PMID:21945607

  4. 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.

  5. Magnetic resonance imaging of ultrasmall superparamagnetic iron oxide-labeled exosomes from stem cells: a new method to obtain labeled exosomes

    PubMed Central

    Busato, Alice; Bonafede, Roberta; Bontempi, Pietro; Scambi, Ilaria; Schiaffino, Lorenzo; Benati, Donatella; Malatesta, Manuela; Sbarbati, Andrea; Marzola, Pasquina; Mariotti, Raffaella

    2016-01-01

    Purpose Recent findings indicate that the beneficial effects of adipose stem cells (ASCs), reported in several neurodegenerative experimental models, could be due to their paracrine activity mediated by the release of exosomes. The aim of this study was the development and validation of an innovative exosome-labeling protocol that allows to visualize them with magnetic resonance imaging (MRI). Materials and methods At first, ASCs were labeled using ultrasmall superparamagnetic iron oxide nanoparticles (USPIO, 4–6 nm), and optimal parameters to label ASCs in terms of cell viability, labeling efficiency, iron content, and magnetic resonance (MR) image contrast were investigated. Exosomes were then isolated from labeled ASCs using a standard isolation protocol. The efficiency of exosome labeling was assessed by acquiring MR images in vitro and in vivo as well as by determining their iron content. Transmission electron microscopy images and histological analysis were performed to validate the results obtained. Results By using optimized experimental parameters for ASC labeling (200 µg Fe/mL of USPIO and 72 hours of incubation), it was possible to label 100% of the cells, while their viability remained comparable to unlabeled cells; the detection limit of MR images was of 102 and 2.5×103 ASCs in vitro and in vivo, respectively. Exosomes isolated from previously labeled ASCs retain nanoparticles, as demonstrated by transmission electron microscopy images. The detection limit by MRI was 3 µg and 5 µg of exosomes in vitro and in vivo, respectively. Conclusion We report a new approach for labeling of exosomes by USPIO that allows detection by MRI while preserving their morphology and physiological characteristics. PMID:27330291

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

    PubMed Central

    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

    2015-01-01

    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. PMID:26582263

  7. 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

    2015-01-01

    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. PMID:26582263

  8. 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

  9. A dual labelling method for measuring uptake of low molecular weight compounds into the pathogenic yeast Candida albicans.

    PubMed

    Ziegelbauer, K

    1998-10-01

    In contrast to other eukaryotic cells the pathogenic yeast Candida albicans is resistant to many structurally unrelated metabolic inhibitors. Reduced permeability due to the cell wall and/or altered plasma membrane composition is thought to be at least partly responsible for this phenomenon. To study the uptake of low molecular weight compounds into C. albicans we developed a dual labelling method. Intact cells, metabolically inactivated cells, spheroplasts or membrane fragments of C. albicans were incubated with various [14C]-labelled compound in the presence of [3H]-labelled water. After separation of cells and supernatant isotope ratios [3H]/[14C] were determined. Quotients of the isotope ratios from cells and supernatant, called enrichment coefficients, were calculated under all four conditions. The enrichment coefficients indicated whether a compound can enter C. albicans cells, is trapped within the cell wall, is enriched in the lipophilic membrane compartment, is actively accumulated or actively exported by multidrug resistance carriers. We used six structurally unrelated compounds to test our method. We found no evidence for a general impermeability of C. albicans.

  10. 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

  11. 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...

  12. 40 CFR 600.115-11 - Criteria for determining the fuel economy label calculation method.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 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...

  13. 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.

  14. Use of stable isotope-labelled cells to identify active grazers of picocyanobacteria in ocean surface waters

    PubMed Central

    Frias-Lopez, Jorge; Thompson, Anne; Waldbauer, Jacob; Chisholm, Sallie W

    2009-01-01

    Prochlorococcus and Synechococcus are the two most abundant marine cyanobacteria. They represent a significant fraction of the total primary production of the world oceans and comprise a major fraction of the prey biomass available to phagotrophic protists. Despite relatively rapid growth rates, picocyanobacterial cell densities in open-ocean surface waters remain fairly constant, implying steady mortality due to viral infection and consumption by predators. There have been several studies on grazing by specific protists on Prochlorococcus and Synechococcus in culture, and of cell loss rates due to overall grazing in the field. However, the specific sources of mortality of these primary producers in the wild remain unknown. Here, we use a modification of the RNA stable isotope probing technique (RNA-SIP), which involves adding labelled cells to natural seawater, to identify active predators that are specifically consuming Prochlorococcus and Synechococcus in the surface waters of the Pacific Ocean. Four major groups were identified as having their 18S rRNA highly labelled: Prymnesiophyceae (Haptophyta), Dictyochophyceae (Stramenopiles), Bolidomonas (Stramenopiles) and Dinoflagellata (Alveolata). For the first three of these, the closest relative of the sequences identified was a photosynthetic organism, indicating the presence of mixotrophs among picocyanobacterial predators. We conclude that the use of RNA-SIP is a useful method to identity specific predators for picocyanobacteria in situ, and that the method could possibly be used to identify other bacterial predators important in the microbial food-web. PMID:19196281

  15. Use of stable isotope-labelled cells to identify active grazers of picocyanobacteria in ocean surface waters.

    PubMed

    Frias-Lopez, Jorge; Thompson, Anne; Waldbauer, Jacob; Chisholm, Sallie W

    2009-02-01

    Prochlorococcus and Synechococcus are the two most abundant marine cyanobacteria. They represent a significant fraction of the total primary production of the world oceans and comprise a major fraction of the prey biomass available to phagotrophic protists. Despite relatively rapid growth rates, picocyanobacterial cell densities in open-ocean surface waters remain fairly constant, implying steady mortality due to viral infection and consumption by predators. There have been several studies on grazing by specific protists on Prochlorococcus and Synechococcus in culture, and of cell loss rates due to overall grazing in the field. However, the specific sources of mortality of these primary producers in the wild remain unknown. Here, we use a modification of the RNA stable isotope probing technique (RNA-SIP), which involves adding labelled cells to natural seawater, to identify active predators that are specifically consuming Prochlorococcus and Synechococcus in the surface waters of the Pacific Ocean. Four major groups were identified as having their 18S rRNA highly labelled: Prymnesiophyceae (Haptophyta), Dictyochophyceae (Stramenopiles), Bolidomonas (Stramenopiles) and Dinoflagellata (Alveolata). For the first three of these, the closest relative of the sequences identified was a photosynthetic organism, indicating the presence of mixotrophs among picocyanobacterial predators. We conclude that the use of RNA-SIP is a useful method to identity specific predators for picocyanobacteria in situ, and that the method could possibly be used to identify other bacterial predators important in the microbial food-web.

  16. CONSTANd : A Normalization Method for Isobaric Labeled Spectra by Constrained Optimization.

    PubMed

    Maes, Evelyne; Hadiwikarta, Wahyu Wijaya; Mertens, Inge; Baggerman, Geert; Hooyberghs, Jef; Valkenborg, Dirk

    2016-08-01

    In quantitative proteomics applications, the use of isobaric labels is a very popular concept as they allow for multiplexing, such that peptides from multiple biological samples are quantified simultaneously in one mass spectrometry experiment. Although this multiplexing allows that peptide intensities are affected by the same amount of instrument variability, systematic effects during sample preparation can also introduce a bias in the quantitation measurements. Therefore, normalization methods are required to remove this systematic error. At present, a few dedicated normalization methods for isobaric labeled data are at hand. Most of these normalization methods include a framework for statistical data analysis and rely on ANOVA or linear mixed models. However, for swift quality control of the samples or data visualization a simple normalization technique is sufficient. To this aim, we present a new and easy-to-use data-driven normalization method, named CONSTANd. The CONSTANd method employs constrained optimization and prior information about the labeling strategy to normalize the peptide intensities. Further, it allows maintaining the connection to any biological effect while reducing the systematic and technical errors. As a result, peptides can not only be compared directly within a multiplexed experiment, but are also comparable between other isobaric labeled datasets from multiple experimental designs that are normalized by the CONSTANd method, without the need to include a reference sample in every experimental setup. The latter property is especially useful when more than six, eight or ten (TMT/iTRAQ) biological samples are required to detect differential peptides with sufficient statistical power and to optimally make use of the multiplexing capacity of isobaric labels. PMID:27302888

  17. 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.

  18. 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. PMID:17630221

  19. 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

  20. 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.

  1. 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. PMID:11208938

  2. 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.

  3. A simple method for the measurement of labelled compound incorporation into cells in culture.

    PubMed

    Suplisson, A; Boissel, J P

    1976-02-10

    A simple method for the measurement of labelled compound incorporation into cells in layer culture was developed. Compared to other methods it proves to spare time and to be more sensitive owing to the fact that cells are not detached from the culture vials until the end of the manipulation as these are dissolved in the scintillation medium together with the cells just before counting.

  4. 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

  5. A fully enzymatic method for site-directed spin labeling of long RNA.

    PubMed

    Lebars, Isabelle; Vileno, Bertrand; Bourbigot, Sarah; Turek, Philippe; Wolff, Philippe; Kieffer, Bruno

    2014-09-01

    Site-directed spin labeling is emerging as an essential tool to investigate the structural and dynamical features of RNA. We propose here an enzymatic method, which allows the insertion of a paramagnetic center at a specific position in an RNA molecule. The technique is based on a segmental approach using a ligation protocol with T4 RNA ligase 2. One transcribed acceptor RNA is ligated to a donor RNA in which a thio-modified nucleotide is introduced at its 5'-end by in vitro transcription with T7 RNA polymerase. The paramagnetic thiol-specific reagent is subsequently attached to the RNA ligation product. This novel strategy is demonstrated by introducing a paramagnetic probe into the 55 nucleotides long RNA corresponding to K-turn and Specifier Loop domains from the Bacillus subtilis tyrS T-Box leader RNA. The efficiency of the coupling reaction and the quality of the resulting spin-labeled RNA were assessed by Mass Spectrometry, Electron Paramagnetic Resonance (EPR) and Nuclear Magnetic Resonance (NMR). This method enables various combinations of isotopic segmental labeling and spin labeling schemes, a strategy that will be of particular interest to investigate the structural and dynamical properties of large RNA complexes by NMR and EPR spectroscopies.

  6. 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.

  7. 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.

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

    PubMed

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

    2013-11-29

    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.

  9. Interactions of 14C-labeled multi-walled carbon nanotubes with soil minerals in water.

    PubMed

    Zhang, Liwen; Petersen, Elijah J; Zhang, Wen; Chen, Yongsheng; Cabrera, Miguel; Huang, Qingguo

    2012-07-01

    Carbon nanotubes are often modified to be stable in the aqueous phase by adding extensive hydrophilic surface functional groups. The stability of such CNTs in water with soil or sediment is one critical factor controlling their environmental fate. We conducted a series of experiments to quantitatively assess the association between water dispersed multi-walled carbon nanotubes (MWCNTs) and three soil minerals (kaolinite, smectite, or shale) in aqueous solution under different sodium concentrations. (14)C-labeling was used in these experiments to unambiguously quantify MWCNTs. The results showed that increasing ionic strength strongly promoted the removal of MWCNTs from aqueous phase. The removal tendency is inversely correlated with the soil minerals' surface potential and directly correlated with their hydrophobicity. This removal can be interpreted by the extended Derjaguin-Landau-Verwey-Overbeek (EDLVO) theory especially for kaolinite and smectite. Shale, which contains large and insoluble organic materials, sorbed MWCNTs the most strongly.

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

    PubMed

    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 B(12) 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 ϵ. The ϵ-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.

  11. 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.

  12. 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.

  13. Label-free method for anti-glucopeptide antibody detection in Multiple Sclerosis

    PubMed Central

    Real-Fernández, Feliciana; Rossi, Giada; Lolli, Francesco; Papini, Anna Maria; Rovero, Paolo

    2015-01-01

    Surface plasmon resonance technique is particularly interesting in immunology because it has the potential to visualize label-free antigen-antibody interactions in real-time, thus enabling antibody detection and monitoring. Herein we release the guidelines for the correct use of a method to detect specific antibodies directly in Multiple Sclerosis patients’ sera using a glucopeptide-based label-free biosensor. The protocol describes the strategy employed for the immobilization of glucopeptide antigen onto a gold sensor chip and the evaluation of the specific binding of serum antibodies to the immobilized antigen. • Label-free method for the real time screening of disease-specific antibodies within a few minutes; • The described protocol employs small quantities of glucopeptide antigen and blood serum samples saving method-cost; • Stability of the immobilized glucopeptide antigen guarantees the regeneration of the surface allowing re-use the immunosensor with high automated throughput. The antibodies detected using the described methodology can be evaluated as biomarkers of Multiple Sclerosis. The SPR detection system is able to characterize antibodies significantly different from those evaluated in the classical enzyme-linked immunosorbent assays (ELISA). PMID:26150982

  14. Label-free method for anti-glucopeptide antibody detection in Multiple Sclerosis.

    PubMed

    Real-Fernández, Feliciana; Rossi, Giada; Lolli, Francesco; Papini, Anna Maria; Rovero, Paolo

    2015-01-01

    Surface plasmon resonance technique is particularly interesting in immunology because it has the potential to visualize label-free antigen-antibody interactions in real-time, thus enabling antibody detection and monitoring. Herein we release the guidelines for the correct use of a method to detect specific antibodies directly in Multiple Sclerosis patients' sera using a glucopeptide-based label-free biosensor. The protocol describes the strategy employed for the immobilization of glucopeptide antigen onto a gold sensor chip and the evaluation of the specific binding of serum antibodies to the immobilized antigen. •Label-free method for the real time screening of disease-specific antibodies within a few minutes;•The described protocol employs small quantities of glucopeptide antigen and blood serum samples saving method-cost;•Stability of the immobilized glucopeptide antigen guarantees the regeneration of the surface allowing re-use the immunosensor with high automated throughput. The antibodies detected using the described methodology can be evaluated as biomarkers of Multiple Sclerosis. The SPR detection system is able to characterize antibodies significantly different from those evaluated in the classical enzyme-linked immunosorbent assays (ELISA). PMID:26150982

  15. Human polyclonal immunoglobulin labelled with technetium-99m via NHS-MAG3: a comparison of radiochemical behavior and biological efficacy with other labelling methods.

    PubMed

    Gano, L; Patrício, L; Marques, E; Cantinho, G; Pena, H; Martins, T; Hnatowich, D J

    1998-05-01

    The aim of this study was to evaluate the radiochemical behavior, biological distribution, and localization in infection sites in mice of a human polyclonal immunoglobulin (HIG) labelled with 99mTc by a novel MAG3-labelling method. The resulting [99mTc]MAG3-HIG was compared with [99mTc]HIG preparations radiolabelled directly via 2-mercaptoethanol (2-Me) or stannous ion (Sn) reduction and indirectly via 2-iminothiolane (2-Im) conjugation. All preparations showed similar UV and radioactivity HPLC profile to that of native HIG except for 2-Im-HIG, which showed aggregates. The stabilities of the label to challenge with cysteine were similar for all the preparations. By nondenaturing SDS-PAGE, all preparations other than MAG3-HIG showed evidence of lower molecular weight fragments. The tissue distribution 4 and 24 h after intravenous administration of the four preparations were compared in mice previously administered with an isolate of Staphylococcus aureus in one thigh. The pharmacokinetics varied among the different preparations. When prepared via 2-Me, Sn, and 2-Im, both blood clearance and urinary excretion were faster than that of labelled MAG3-HIG. The absolute uptake in the infected thigh at 24 h was significantly higher for HIG labelled via MAG3 and 2-Me vs. the remaining methods. The infected thigh/normal thigh radioactivity ratios were similar at both time points for labelled HIG prepared via 2-Me, 2-Im, and NHS-MAG, methods but was significantly lower at 24 h for HIG prepared via Sn. The radioactive HPLC profiles of serum at 4 and 24 h were similar to that of the radiolabelled injectates. Based on these data we conclude that each radiolabelled HIG preparation studied showed increased localization in infectious foci although [99Tc]MAG3-HIG showed superior radiochemical and biological characteristics under the conditions of this investigation.

  16. Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids. Part Two: Colorigenic methods and comparison with chemiluminescent methods.

    PubMed

    Rihn, B; Bottin, M C; Coulais, C; Martinet, N

    1995-06-01

    The diagnosis of genetic infections and cancerous diseases is carried out more and more often at a molecular level using Southern's technique which is based on the use of 32P-labelled DNA. In order to circumvent the risks and rapid decrease in radioactivity associated with these latter techniques, new colorigenic methods have been developed. In this work, we describe the use of dTTP analogues (digoxigenin-dUTP and biotin-dUTP) for the labelling of probes and detection of target DNA. Using digoxigenin-11-dUTP, 0.1 aM of a 561 bp target DNA was detected by using a modified Southern procedure. The reliability and the high sensitivity of such methods make them a good tool for DNA investigation in research as well as in testing laboratories.

  17. 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.

  18. A general method for the covalent labeling of fusion proteins with small molecules in vivo.

    PubMed

    Keppler, Antje; Gendreizig, Susanne; Gronemeyer, Thomas; Pick, Horst; Vogel, Horst; Johnsson, Kai

    2003-01-01

    Characterizing the movement, interactions, and chemical microenvironment of a protein inside the living cell is crucial to a detailed understanding of its function. Most strategies aimed at realizing this objective are based on genetically fusing the protein of interest to a reporter protein that monitors changes in the environment of the coupled protein. Examples include fusions with fluorescent proteins, the yeast two-hybrid system, and split ubiquitin. However, these techniques have various limitations, and considerable effort is being devoted to specific labeling of proteins in vivo with small synthetic molecules capable of probing and modulating their function. These approaches are currently based on the noncovalent binding of a small molecule to a protein, the formation of stable complexes between biarsenical compounds and peptides containing cysteines, or the use of biotin acceptor domains. Here we describe a general method for the covalent labeling of fusion proteins in vivo that complements existing methods for noncovalent labeling of proteins and that may open up new ways of studying proteins in living cells.

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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. PMID:22771841

  6. 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.

  7. 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

    2015-08-01

    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.

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

    PubMed

    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 [(125)I]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 [(125)I]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 [(125)I]iodide on the amount of [(125)I]iodine monochloride formed, (b) the influence of the specific radioactivity of [(125)I]iodide on the amount of [(125)I]iodine monochloride formed, and (c) the influence of the specific radioactivity of [(125)I]iodide on the number of millicuries needed for labelling to a desired extent.

  9. 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

  10. 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.

  11. 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

  12. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...) The manufacturer or private labeler may include the ENERGY STAR logo on the bottom right corner of the... may add the ENERGY STAR logo to labels on certified covered products; such manufacturers may add the ENERGY STAR logo to labels only on those covered products that are contemplated by the Memorandum...

  13. 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...

  14. 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...

  15. 16 CFR 305.11 - Labeling for refrigerators, refrigerator-freezers, freezers, dishwashers, clothes washers, water...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... bottom). Depth is variable but should follow closely the prototype labels appearing at the end of this... 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...

  16. .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.

  17. 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.

  18. 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. PMID:21724183

  19. Rapid triple-labeling method combining in situ hybridization and double immunocytochemistry.

    PubMed

    Lopez-Sanchez, Carmen; Garcia-Martinez, Virginio; Lawson, Aaron; Chapman, Susan C; Schoenwolf, Gary C

    2004-06-01

    A new, rapid method is described for combining in situ hybridization and immunocytochemistry to define cell populations and to map three-dimensional movements of groups of labeled cells within developing chick embryos. The method allows fluorescently labeled cells to be followed in living embryos and subsequently detected as a permanent reaction product for detailed three-dimensional analysis by immunocytochemistry in histological serial sections. Cell identity can be ascertained using a specific riboprobe and in situ hybridization. With this approach, the movements of two groups of cells can be mapped simultaneously (using two different fluorescent trackers and, subsequently, two different chromogens for immunocytochemistry) to analyze relative movements within an embryo, and when combined with in situ hybridization with a specific riboprobe for cell identity, allows fate mapping studies to be conducted using molecular criteria, rather than solely at morphological/positional criteria. The improved method enables the investigator to extract substantially more information from individual embryos, maximizing the results obtained from labor-intensive fate mapping studies.

  20. 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.

  1. Comparison of quantification methods of 111In-labelled platelet deposition in peripheral bypass grafts.

    PubMed

    Mortelmans, L; Verbruggen, A; De Bakker, C; Vandecruys, A; Joosten, J; Nevelsteen, A; Noyez, L; Verstraete, M; Vermylen, J; De Roo, M

    1987-04-01

    The action of antithrombotic drugs can be evaluated by measuring the deposition of 111In-labelled platelets on peripheral bypass grafts several days after injection. This evaluation can be performed qualitatively (visual interpretation on the daily images) or quantitatively. Four different methods which calculate the ratio of platelet uptake with a reference region are compared: two methods use a gamma camera and two a detector. A blood sample or the region under the sternal angle are used as reference. The daily ratio of the counts, recorded by a gamma camera in a region of interest covering the graft, and the blood radioactivity interpolated from a platelet survival curve appears to be the most reliable method. The information of all the ratios can be combined in a single thrombogenicity index which reflects the daily rise of a linear or exponential regression versus time. PMID:3588323

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

    PubMed

    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.

  3. 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

  4. 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.

  5. 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.

  6. New silver-gold intensification method of diaminobenzidine for double-labeling immunoelectron microscopy.

    PubMed

    Dobó, Endre; Takács, Virág T; Gulyás, Attila I; Nyiri, Gábor; Mihály, András; Freund, Tamás F

    2011-03-01

    The available methods for double-labeling preembedding immunoelectron microscopy are highly limited because not only should the ultrastructure be preserved, but also the different antigens should be visualized by reaction end products that can be clearly distinguished in gray-scale images. In these procedures, one antigen is detected with 3,3'-diaminobenzidine (DAB) chromogen, resulting in a homogeneous deposit, whereas the other is labeled with either a gold-tagged immunoreagent, or DAB polymer, on the surface of which metallic silver is precipitated. The detection of the second antigen is usually impeded by the first, leading to false-negative results. The authors aimed to diminish this hindrance by a new silver intensification technique of DAB polymer, which converts the deposit from amorphous to granular. The method includes three major postdevelopmental steps: (1) treatment of nickel-enhanced DAB with sulfide, (2) silver deposition in the presence of hydroquinone under acidic conditions, and (3) precious metal replacement with gold thiocyanate. This new sulfide-silver-gold intensification of DAB (SSGI) allows a subsequent detection of other antigens using DAB. In conclusion, the new technique loads fine gold particles onto the DAB deposit at a very low background level, thereby allowing a reliable discernment between the elements stained for the two antigens at the ultrastructural level. PMID:21378280

  7. 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. PMID:21128590

  8. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the neck. The fiber content... another conspicuous and readily accessible label or labels on the inside or outside of the garment. On...

  9. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the neck. The fiber content... another conspicuous and readily accessible label or labels on the inside or outside of the garment. On...

  10. 16 CFR 300.5 - Required label and method of affixing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the neck. The fiber content... another conspicuous and readily accessible label or labels on the inside or outside of the garment. On...

  11. 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.

  12. Analytical methods for detection of gluten in food--method developments in support of food labeling legislation.

    PubMed

    Haraszi, Reka; Chassaigne, Hubert; Maquet, Alain; Ulberth, Franz

    2011-01-01

    The current essential therapy of celiac disease is a strict adherence to a gluten-free diet. Besides food products that are naturally gluten-free, "very low gluten" and "gluten-free" bakery products have become available. The availability of immunochemical and other analytical methods to determine gluten markers in foods is of utmost importance to ensure the well being of gluten-sensitive individuals. The aim of this review was to evaluate if currently available methodologies are suitable to meet the requirements of food labeling standards for individual gluten source declaration, in order to achieve policy objectives. Codex Alimentarius and European Union (EU) legislation and gluten detection methodologies applicable at present have been summarized and compared. In 2009, the European Commission issued Regulation No. 41/2009 concerning the composition and labeling of foodstuffs suitable for people intolerant to gluten. This review constitutes a basis to investigate the possibility to develop a proteomic-based method for the specific detection of gluten-containing cereals in food products, especially at or around the limits specified in EU legislation.

  13. 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.

  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. Evaluation of methods for detection of fluorescence labeled subcellular objects in microscope images

    PubMed Central

    2010-01-01

    Background Several algorithms have been proposed for detecting fluorescently labeled subcellular objects in microscope images. Many of these algorithms have been designed for specific tasks and validated with limited image data. But despite the potential of using extensive comparisons between algorithms to provide useful information to guide method selection and thus more accurate results, relatively few studies have been performed. Results To better understand algorithm performance under different conditions, we have carried out a comparative study including eleven spot detection or segmentation algorithms from various application fields. We used microscope images from well plate experiments with a human osteosarcoma cell line and frames from image stacks of yeast cells in different focal planes. These experimentally derived images permit a comparison of method performance in realistic situations where the number of objects varies within image set. We also used simulated microscope images in order to compare the methods and validate them against a ground truth reference result. Our study finds major differences in the performance of different algorithms, in terms of both object counts and segmentation accuracies. Conclusions These results suggest that the selection of detection algorithms for image based screens should be done carefully and take into account different conditions, such as the possibility of acquiring empty images or images with very few spots. Our inclusion of methods that have not been used before in this context broadens the set of available detection methods and compares them against the current state-of-the-art methods for subcellular particle detection. PMID:20465797

  16. (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. PMID:27140906

  17. (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.

  18. An optical method to assess water clarity in coastal waters.

    PubMed

    Kulshreshtha, Anuj; Shanmugam, Palanisamy

    2015-12-01

    Accurate estimation of water clarity in coastal regions is highly desired by various activities such as search and recovery operations, dredging and water quality monitoring. This study intends to develop a practical method for estimating water clarity based on a larger in situ dataset, which includes Secchi depth (Z sd ), turbidity, chlorophyll and optical properties from several field campaigns in turbid coastal waters. The Secchi depth parameter is found to closely vary with the concentration of suspended sediments, vertical diffuse attenuation coefficient K d (m(-1)) and beam attenuation coefficient c (m(-1)). The optical relationships obtained for the selected wavelengths (i.e. 520, 530 and 540 nm) exhibit an inverse relationship between Secchi depth and the length attenuation coefficient (1/(c + K d )). The variation in Secchi depth is expressed in terms of undetermined coupling coefficient which is composed of light penetration factor (expressed by z(1%)K d (λ)) and a correction factor (ξ) (essentially governed by turbidity of the water column). This method of estimating water clarity was validated using independent in situ data from turbid coastal waters, and its results were compared with those obtained from the existing methods. The statistical analysis of the measured and the estimated Z sd showed that the present method yields lower error when compared to the existing methods. The spatial structures of the measured and predicted Z sd are also highly consistent with in situ data, which indicates the potential of the present method for estimating the water clarity in turbid coastal and associated lagoon waters.

  19. Intein Applications: From Protein Purification and Labeling to Metabolic Control Methods*

    PubMed Central

    Wood, David W.; Camarero, Julio A.

    2014-01-01

    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. PMID:24700459

  20. An economical method for (15)N/(13)C isotopic labeling of proteins expressed in Pichia pastoris.

    PubMed

    Rodriguez, E; Krishna, N R

    2001-07-01

    We report a new and cost-effective approach to prepare (15)N/(13)C labeled proteins for NMR using the Pichia pastoris expression system. Four protocols (P1 to P4) were defined and compared using recombinant Ovine interferon-tau (rOvIFN-tau). Our results demonstrate that in order to get full incorporation of (15)N and (13)C, the isotopes are not totally required during the initial growth phase of P. pastoris culture. The addition of small amounts of (15)N and (13)C compounds 6 h prior to the methanol induction phase is sufficient to obtain 99% incorporation of heavy isotopes into the protein. Our optimized protocol P4 is two-thirds less costly than the classical method using (15)N and (13)C isotopes during the entire growth phase.

  1. Immunocytochemical localization of water-soluble glycoproteins, including group 1 allergen, in pollen of ryegrass, Lolium perenne, using ferritin-labelled antibody.

    PubMed

    Vithanage, H I; Howlett, B J; Jobson, S; Knox, R B

    1982-11-01

    The cellular sites of the glycoproteins Group 1 allergen (glycoprotein 1) and Antigen A (glycoprotein 2) in mature ryegrass pollen have been investigated by immunoelectron microscopy. Radioimmunoassays confirm previous findings of cross-reactivity between the purified glycoprotein antigens at the high immunoglobulin G (IgG) concentrations used for localization. Freeze-drying of anthers followed by anhydrous processing has been employed because of the water solubility and mobility of the glycoproteins. A double-embedding technique has been developed. This involves, first, embedding anthers in the water-soluble plastic resin JB-4, sectioning and incubating in ferritin-labelled antisera by the indirect method. The sections are then embedded in Spurr's resin for ultra-thin sectioning. Both glycoproteins are found in the following sites: (1) exine and intine wall layers; (2) pollen cytoplasm; (3) the orbicules and anther loculus; and (4) the anther cuticle. In the exine arcades and surface and in the anther loculus, the ferritin label is bound to pollenkitt. The finding that the glycoproteins are in similar sites is predictable in view of the cross-specificity of the antisera. The extent of antibody penetration of the plastic sections has been examined; labelling is confined to cut grains and absent from intact grains.

  2. Label-Free and High-Throughput Detection of Protein Microarrays by Oblique-Incidence Reflectivity Difference Method

    NASA Astrophysics Data System (ADS)

    Wang, Xu; Lu, Heng; Wen, Juan; Yuan, Kun; LÜ, Hui-Bin; Jin, Kui-Juan; Zhou, Yue-Liang; Yang, Guo-Zhen

    2010-10-01

    We label-free detected the biological process of preparing a microarray that includes 400 spots of mouse immunoglobulin G (IgG) as well as the specific hybridization between mouse IgG and goat anti-mouse IgG by an oblique-incidence reflectivity difference (OI-RD) method. The detection results after each process including printing, washing, blocking, and hybridization, demonstrate that the OI-RD method can trace the preparation process of a microarray and detect the specific hybridization between antigens and antibodies. OI-RD is a promising method for label-free and high-throughput detection of biological microarrays.

  3. 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.

  4. 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).…

  5. 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…

  6. 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. PMID:2497436

  7. 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.

  8. Immunoscintigraphy with indium-111 labeled monoclonal antibodies: The importance of a good display method

    SciTech Connect

    Liehn, J.C.; Hannequin, P.; Nasca, S.; Lebrun, D.; Fernandez-Valoni, A.; Valeyre, J. )

    1989-03-01

    A major drawback of In-111-labeled monoclonal antibodies (MoAb) is the presence of intense liver, renal, and bone marrow nonspecific activity. This makes the display of the images hardly optimal and their visual interpretation difficult. In this study, the intrinsic color scale (which consists of selecting the limits of the color scale as the highest and the lowest pixel value of the image) was compared to a new, simple algorithm for the determination of the limits of the color scale. This algorithm was based on the count density in the iliac crest areas. OC-125 or anti-CEA In-111 MoAb F(ab')2 fragments were used in 32 patients with suspected recurrence of ovarian (19 patients) or colorectal cancer (13 patients). Final diagnosis was assessed by surgery (21 patients), biopsy (five patients), or followup (six patients). A 10-minute abdomino-pelvic anterior view was recorded two days after injection. These views are displayed using the two methods and interpreted by two observers. Using their responses in each quadrant of the pelvis, the authors calculated two ROC curves. The comparison of the ROC curves showed better performances for the new method. For example, for the same specificity (73%), the sensitivity of the new method was significantly better (78% versus 68%). This result confirmed the importance of a good methodology for displaying immunoscintigraphic images.

  9. 16 CFR 303.15 - Required label and method of affixing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., resale and until sold and delivered to the ultimate consumer. (b) Each textile fiber product with a neck must have a label disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the...

  10. 16 CFR 303.15 - Required label and method of affixing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., resale and until sold and delivered to the ultimate consumer. (b) Each textile fiber product with a neck must have a label disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the...

  11. 16 CFR 303.15 - Required label and method of affixing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., resale and until sold and delivered to the ultimate consumer. (b) Each textile fiber product with a neck must have a label disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the...

  12. 16 CFR 303.15 - Required label and method of affixing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ..., resale and until sold and delivered to the ultimate consumer. (b) Each textile fiber product with a neck must have a label disclosing the country of origin affixed to the inside center of the neck midway between the shoulder seams or in close proximity to another label affixed to the inside center of the...

  13. 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

  14. Label-Free Optical Method for Quantifying Molecular Transport Across Cellular Membranes In Vitro.

    PubMed

    Sharifian Gh, Mohammad; Wilhelm, Michael J; Dai, Hai-Lung

    2016-09-01

    We demonstrate a nonlinear optical method for the label-free quantification of membrane transport rates of small/medium size molecules in living cells. Specifically, second-harmonic generation (SHG) laser scattering permits surface-specific characterization of transport across membranes. Unfortunately, most biologically relevant molecules are SHG-inactive. In the interest of extending this methodology for characterizing transport of any molecule, we monitor the SHG produced from an SHG-active reference molecule, in the presence of an SHG-inactive target molecule-of-interest as both molecules compete to cross a membrane. Of significance, the SHG-inactive target transport rate can be deduced as a perturbation in the measured transport rate of the reference. As proof-of-principle, we examine competitive transport of the strongly SHG-active cation, malachite green (MG), in the presence of a weakly SHG-active dication, propidium (Pro), across the outer-membrane protein channels in living bacteria. Comparison of the extracted and directly measured Pro transport rates validates the effectiveness of the method. PMID:27518496

  15. Water-soluble poly(2,7-dibenzosilole) as an ultra-bright fluorescent label for antibody-based flow cytometry.

    PubMed

    Wang, Xin; Hu, Yi-Zhen; Chen, Aimei; Wu, Yexin; Aggeler, Robert; Low, Quentin; Kang, Hee Chol; Gee, Kyle R

    2016-03-14

    A series of novel water-soluble PEGylated dibenzosilole-based conjugated polymers were prepared as ultra-bright fluorescent labels for biomolecules. Due to their superior solubility and brightness, antibody conjugates labeled with functionalized polymers showed significantly enhanced signal and sensitivity relative to traditional fluorophores in functional flow cytometry applications. PMID:26888307

  16. 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

  17. 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.

  18. Label fusion in atlas-based segmentation using a selective and iterative method for performance level estimation (SIMPLE).

    PubMed

    Langerak, Thomas Robin; van der Heide, Uulke A; Kotte, Alexis N T J; Viergever, Max A; van Vulpen, Marco; Pluim, Josien P W

    2010-12-01

    In a multi-atlas based segmentation procedure, propagated atlas segmentations must be combined in a label fusion process. Some current methods deal with this problem by using atlas selection to construct an atlas set either prior to or after registration. Other methods estimate the performance of propagated segmentations and use this performance as a weight in the label fusion process. This paper proposes a selective and iterative method for performance level estimation (SIMPLE), which combines both strategies in an iterative procedure. In subsequent iterations the method refines both the estimated performance and the set of selected atlases. For a dataset of 100 MR images of prostate cancer patients, we show that the results of SIMPLE are significantly better than those of several existing methods, including the STAPLE method and variants of weighted majority voting. PMID:20667809

  19. 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

  20. 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.

  1. Interaction of ethanol with 111In-labelled membranes: evaluation by the perturbed angular correlation-sum peak ratio method.

    PubMed

    Jay, M; Woodward, M A

    1985-08-01

    The interaction of ethanol with erythrocyte ghosts and vesicles composed of brain lipid extracts labelled with indium-111 was studied using the sum peak ratio method of perturbed angular correlation measurements. Membranes from animals that were fed diets containing ethanol for 10 days demonstrated resistance to the decrease in sum peak ratio values observed in control animals. Thus, repeated administration of ethanol induces changes in the properties of biological membranes, possibly by altering phospholipid composition, which is reflected in the anisotropy of membrane-associated 111In-labelled nuclei as measured by sum peak ratios.

  2. Analysis of the interaction of reserpine with actin by the photoaffinity labelling method.

    PubMed

    Ohmi, K; Nakamura, S

    1991-08-14

    The interaction of reserpine with one of the cytoskeletal proteins, actin, was analyzed by the photoaffinity labelling method using [3H]reserpine. Reserpine bound sufficiently to G- or oligomeric actin, but hardly to F-actin under the same experimental conditions. This result could be explained if reserpine binds to a specific region of the G-actin molecule that is responsible for actin-actin interactions. It is concordant with this idea that [3H]reserpine bound only to specific proteolytic fragments of actin. When reserpine was mixed with crude extracts of two kinds of tissues, chicken gizzard smooth muscle and bovine adrenal medulla, it bound to the 42 kDa protein of sodium dodecyl sulfate-polyacrylamide gel electrophoresis in both cases. Its molecular size and abundance suggest strongly that this 42 kDa protein is actin. The binding of reserpine to actin was inhibited by dopamine in a dose-dependent manner. These results suggest that actin could be one of the target molecules of reserpine.

  3. A broadband capacitive sensing method for label-free bacterial LPS detection.

    PubMed

    Rydosz, Artur; Brzozowska, Ewa; Górska, Sabina; Wincza, Krzysztof; Gamian, Andrzej; Gruszczynski, Slawomir

    2016-01-15

    In this paper, the authors present a new type of highly sensitive label-free microwave sensor in a form of interdigital capacitor coated with T4 bacteriophage gp37 adhesin. The adhesin binds Escherichia coli B (E. coli B) by precise recognizing its bacterial host lipopolysaccharide (LPS). The C-terminal part of the adhesin consists of the receptor-binding amino acid residues which are involved in a specific interaction with two terminal glucose residues of the bacterial LPS. The change of the sensors' capacitance and conductance as a subject to LPS presence is an indicator of the detection. The measurements in the frequency range of 0-3GHz utilizing vector network analyzer have been carried out at different concentrations to verify experimentally the proposed method. The measured capacitance change between the reference and the biofunctionalized sensor equals 15% in the entire frequency range and the measured conductance change exceeds 19%. The changes of both parameters can be used as good indicators of the LPS detection. The selectivity has been confirmed by the ELISA experiments and tested by sensor measurements with lipopolysaccharide (LPS) from E. coli B, E. coli 056, E. coli 0111, Pseudomonas aeruginosa NBRC 13743 and Hafnia alvei 1185. PMID:26339930

  4. 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

  5. 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.

  6. 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

  7. Dual-labeling method for electron microscopy to characterize synaptic connectivity using genetically encoded fluorescent reporters in Drosophila.

    PubMed

    Tanaka, Nobuaki K; Dye, Louis; Stopfer, Mark

    2011-01-15

    Light and electron microscopy (LM and EM) both offer important advantages for characterizing neuronal circuitry in intact brains: LM can reveal the general patterns neurons trace between brain areas, and EM can confirm synaptic connections between identified neurons within a small area. In a few species, genetic labeling with fluorescent proteins has been used with LM to visualize many kinds of neurons and to analyze their morphologies and projection patterns. However, combining these large-scale patterns with the fine detail available in EM analysis has been a technical challenge. To analyze the synaptic connectivity of neurons expressing fluorescent markers with EM, we developed a dual-labeling method for use with pre-embedded brains. In Drosophila expressing genetic labels and also injected with markers we visualized synaptic connections among two populations of neurons in the AL, one of which has been shown to mediate a specific function, odor evoked neural oscillation.

  8. Dual-labeling method for electron microscopy to characterize synaptic connectivity using genetically encoded fluorescent reporters in Drosophila

    PubMed Central

    Tanaka, Nobuaki K.; Dye, Louis; Stopfer, Mark

    2010-01-01

    Light and electron microscopy (LM and EM) both offer important advantages for characterizing neuronal circuitry in intact brains: LM can reveal the general patterns neurons trace between brain areas, and EM can confirm synaptic connections between identified neurons within a small area. In a few species, genetic labeling with fluorescent proteins has been used with LM to visualize many kinds of neurons and to analyze their morphologies and projection patterns. However, combining these large-scale patterns with the fine detail available in EM analysis has been a technical challenge. To analyze the synaptic connectivity of neurons expressing fluorescent markers with EM, we developed a dual-labeling method for use with pre-embedded brains. In Drosophila expressing genetic labels and also injected with markers we visualized synaptic connections among two populations of neurons in the AL, one of which has been shown to mediate a specific function, odor evoked neural oscillation. PMID:21074556

  9. 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.

  10. 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,…

  11. 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

  12. Method for the Determination of ¹⁵N Incorporation Percentage in Labeled Peptides and Proteins.

    PubMed

    Kilpatrick, Eric L

    2016-01-01

    Use of labeled (15)N proteins and peptides as internal standards in isotope-dilution mass spectrometry for the quantification of proteins has been increasing and is now accepted as a gold standard for this analysis. As a necessary reagent in this process, stable heavy isotope-labeled internal standards must be rigorously characterized in a number of ways including identity, concentration, purity, and structure. Additionally, the degree of the incorporation of the heavy isotope is a critical feature to consider. For proteins that are (15)N labeled, the percentage of incorporation is a valid measurement used to assess the fitness-to-purpose of the material. This measurement should be objective, repeatable, and based on empirical analysis. One means of assigning this value is to compare a mass spectrum of the isotopic profile of a peptide against a series of theoretical profiles containing different enrichment rates. This comparison can be made using the Pearson product-moment correlation coefficient (r) to find the best match between the empirical and theoretical profiles. Theoretical profiles can be generated using probability multinomial analysis but are computationally intensive and require the use of computers for practical use. The method described in this chapter describes the development and use of a computer program to calculate the percentage of (15)N enrichment of a labeled internal standard. Additionally, methods will be described for the empirical determination of an isotopic profile using a variety of mass spectrometry techniques.

  13. 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

  14. The beauty of being (label)-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics.

    PubMed

    Vowinckel, Jakob; Capuano, Floriana; Campbell, Kate; Deery, Michael J; Lilley, Kathryn S; Ralser, Markus

    2013-01-01

    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

  15. 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. PMID:25960325

  16. 10 CFR 431.106 - Uniform test method for the measurement of energy efficiency of commercial water heaters and hot...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... and Unfired Hot Water Storage Tanks Test Procedures § 431.106 Uniform test method for the measurement..., pursuant to EPCA, you are measuring the thermal efficiency or standby loss, or both, of a storage or... procedures in subsection labeledMethod of Test” of With these additional stipulations Gas-fired Storage...

  17. 10 CFR 431.106 - Uniform test method for the measurement of energy efficiency of commercial water heaters and hot...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... and Unfired Hot Water Storage Tanks Test Procedures § 431.106 Uniform test method for the measurement..., pursuant to EPCA, you are measuring the thermal efficiency or standby loss, or both, of a storage or... procedures in subsection labeledMethod of Test” of With these additional stipulations Gas-fired Storage...

  18. 10 CFR 431.106 - Uniform test method for the measurement of energy efficiency of commercial water heaters and hot...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... and Unfired Hot Water Storage Tanks Test Procedures § 431.106 Uniform test method for the measurement..., pursuant to EPCA, you are measuring the thermal efficiency or standby loss, or both, of a storage or... procedures in subsection labeledMethod of Test” of With these additional stipulations Gas-fired Storage...

  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

    2015-04-01

    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. Characterization of Metabolically Quiescent Leishmania Parasites in Murine Lesions Using Heavy Water Labeling

    PubMed Central

    Kloehn, Joachim; Saunders, Eleanor C.; O’Callaghan, Sean; Dagley, Michael J.; McConville, Malcolm J.

    2015-01-01

    Information on the growth rate and metabolism of microbial pathogens that cause long-term chronic infections is limited, reflecting the absence of suitable tools for measuring these parameters in vivo. Here, we have measured the replication and physiological state of Leishmania mexicana parasites in murine inflammatory lesions using 2H2O labeling. Infected BALB/c mice were labeled with 2H2O for up to 4 months, and the turnover of parasite DNA, RNA, protein and membrane lipids estimated from the rate of deuterium enrichment in constituent pentose sugars, amino acids, and fatty acids, respectively. We show that the replication rate of parasite stages in these tissues is very slow (doubling time of ~12 days), but remarkably constant throughout lesion development. Lesion parasites also exhibit markedly lower rates of RNA synthesis, protein turnover and membrane lipid synthesis than parasite stages isolated from ex vivo infected macrophages or cultured in vitro, suggesting that formation of lesions induces parasites to enter a semi-quiescent physiological state. Significantly, the determined parasite growth rate accounts for the overall increase in parasite burden indicating that parasite death and turnover of infected host cells in these lesions is minimal. We propose that the Leishmania response to lesion formation is an important adaptive strategy that minimizes macrophage activation, providing a permissive environment that supports progressive expansion of parasite burden. This labeling approach can be used to measure the dynamics of other host-microbe interactions in situ. PMID:25714830

  1. Ultrasensitive and rapid screening of mercury(II) ions by dual labeling colorimetric method in aqueous samples and applications in mercury-poisoned animal tissues.

    PubMed

    Deng, Yi; Wang, Xin; Xue, Feng; Zheng, Lei; Liu, Jian; Yan, Feng; Xia, Fan; Chen, Wei

    2015-04-01

    Rapid and ultrasensitive detection of trace heavy metal mercury(II) ions (Hg(2+)) are of significant importance due to the induced serious risks for environment and human health. This presented article reports the gold nanoparticle-based dual labeling colorimetric method (Dual-COLO) for ultrasensitive and rapid detection of Hg(2+) using the specific thymine-Hg(2+)-thymine (T-Hg(2+)-T) as recognition system and the dual labeling strategy for signal amplification. Both qualitative and quantitative detections of Hg(2+) are achieved successfully in aqueous samples. More importantly, the achieved detection limit of 0.005 ng mL(-1) (0.025 nM) without any instruments is very competitive to other rapid detection methods even ICP-MS based methods. This Dual-COLO method is also applied directly for real water sample monitoring and, more importantly, applied in analysis of mercury poisoned animal tissues and body fluidic samples, indicating a potentially powerful and promising tool for environmental monitoring and food safety control.

  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. 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).

  4. 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.

  5. 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.

  6. 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

  7. Comparison of oxine and tropolone methods for labeling human platelets with indium-111

    SciTech Connect

    Kotze, H.F.; Heyns, A.D.; Loetter, M.G.P.; Pieters, H.; Roodt, J.P.; Sweetlove, M.A.; Badenhorst, P.N. )

    1991-01-01

    The effect of the chelates oxine and tropolone, used to label platelets, on the kinetics of indium-111-({sup 111}In) labeled platelets was studied in twelve normal human subjects. Autologous platelets were labeled either in saline with {sup 111}In-oxine or in plasma with {sup 111}In-tropolone. Mean platelet lifespan was estimated by fitting the disappearance curve of platelets from the circulation to the multiple hit and other mathematical models. The in vivo distribution of platelets was quantitatively imaged with a scintillation camera. The in vivo recovery of {sup 111}In-oxine and {sup 111}In-tropolone did not differ, and the mean platelet lifespan was also similar ({sup 111}In-oxine: 230 +/- 29 hr; {sup 111}In-tropolone: 226 +/- 13 hr). At equilibrium (90 min after reinjection of labeled platelets) and at the end of platelet lifespan, {sup 111}In-oxine and {sup 111}In-tropolone radioactivities in the spleen and liver were similar. These results demonstrate that the results of kinetics measured with {sup 111}In-oxine or {sup 111}In-tropolone do not differ significantly.

  8. 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.

  9. 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.

  10. Four-day multimedia diet records underestimate energy needs in middle-aged and elderly women as determined by doubly-labeled water.

    PubMed

    Kaczkowski, C H; Jones, P J; Feng, J; Bayley, H S

    2000-04-01

    Systematic problems exist in the quantification of food intake in populations using traditional self-reported measures. The objective of this study was to determine the effectiveness of an innovative multimedia diet record (MMDR) for dietary energy intake assessment. Dietary intake was estimated by combining the use of a microcassette tape recorder and 35-mm camera in 53 women whose ages ranged from 50 to 93 y (64.9 +/- 11.3 y), with body weights of 62.4 +/- 12.2 kg and body mass indexes (BMI) of 24.4 +/- 4.0 kg/m(2). Using household measures, subjects voice-recorded and photographed all food and beverages consumed for four consecutive days. A two-point doubly-labeled water (DLW) method was used over 13 d to calculate carbon dioxide production, total body water, and subsequently, total energy expenditure (TEE) through the use of a food quotient. Mean body weight did not change between d 1 and 14. TEE and reported energy intake were compared using MMDR. Mean reported energy intakes 7.5 +/- 1.9 MJ/d (1774 +/- 476 kcal/d) were lower (P < 0.01) than TEE by 10.4 +/- 3.1 MJ/d (2477 +/- 736 kcal/d), indicating underreporting of food intake. Reporting accuracy (reported energy intake/TEE' 100%) was 76.0 +/- 22.9%. Mean energy expenditure (MJ/d), as determined by doubly-labeled water, was higher (P < 0.01) in each stratified age range when compared to reported energy intake by MMDR. There were no significant differences in reporting accuracy among the stratified age groups. Using the MMDR method, this population of weight-stable women underreported their food intakes compared to their determined energy expenditure estimated by DLW.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. Accumulation and fate of green fluorescent labeled Escherichia coli in laboratory-scale drinking water biofilters.

    PubMed

    Li, J; McLellan, S; Ogawa, S

    2006-09-01

    Biological filters combining microbial activity and rapid sand filtration are used in drinking water treatment plants for enhanced biodegradable organic matters (BOM) removal. Biofilms formed on filter media comprised of bacteria enclosed in a polymeric matrix are responsible for the adsorption of BOM and attachment of planktonic microorganisms. This study investigated the removal of Escherichia coli cells injected into laboratory-scale biofilters and the role of biofilm in retaining the injected E. coli. Green fluorescent protein was used as a specific marker to detect and quantify E. coli in the biofilms. About 35% of the total injected E. coli cells were observed in the filter effluents, when initial cell concentrations were measured at 7.4 x 10(6) CFU/mL and 1.6 x 10(7) CFU/mL in two separate experiments. The results from real-time PCR and plate count analysis indicated that 95% of the E. coli retained inside the filters were either non-viable or could not be recovered by colony counting techniques. Injected cells were unevenly distributed inside the filter with more than 70% located at the top 1/5 of the filter. Images obtained from an epifluorescent microscope showed that E. coli cells were embedded inside the biofilm matrix and presented mainly as microcolonies intertwined with other microorganisms, which was consistent with findings from standard plate count methods and qPCR.

  16. 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.

  17. Evaluation of chemical labeling methods for identifying functional arginine residues of proteins by mass spectrometry.

    PubMed

    Wanigasekara, Maheshika S K; Chowdhury, Saiful M

    2016-09-01

    Arginine residues undergo several kinds of post-translational modifications (PTMs). These PTMs are associated with several inflammatory diseases, such as rheumatoid arthritis, atherosclerosis, and diabetes. Mass spectrometric studies of arginine modified proteins and peptides are very important, not only to identify the reactive arginine residues but also to understand the tandem mass spectrometry behavior of these peptides for assigning the sequences unambiguously. Herein, we utilize tandem mass spectrometry to report the performance of two widely used arginine labeling reagents, 1,2-cyclohexanedione (CHD) and phenylglyoxal (PG) with several arginine containing peptides and proteins. Time course labeling studies were performed to demonstrate the selectivity of the reagents in proteins or protein digests. Structural studies on the proteins were also explored to better understand the reaction sites and position of arginine residues. We found CHD showed better labeling efficiencies compared to phenylglyoxal. Reactive arginine profiling on a purified albumin protein clearly pointed out the cellular glycation modification site for this protein with high confidence. We believe these detailed mass-spectrometric studies will provide significant input to profile reactive arginine residues in large-scale studies; therefore, targeted proteomics can be performed to the short listed reactive sites for cellular arginine modifications. PMID:27543028

  18. 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.

  19. Label-acquired magnetorotation as a signal transduction method for protein detection: aptamer-based detection of thrombin

    PubMed Central

    Hecht, Ariel; Akshay Kumar, Anand; Kopelman, Raoul

    2011-01-01

    This paper presents a new signal transduction method, called Label-Acquired Magnetorotation (LAM), for the measurement of proteins in solution. We demonstrate the use of LAM to detect the protein thrombin using aptamers, with an LOD (limit of detection) of 300 pM. LAM is modeled after a sandwich assay, with a 10 µm nonmagnetic “mother” sphere as the capture component, and with 1 µm magnetic “daughter” beads as the labels. The protein-mediated attachment of daughter beads to the mother sphere forms a rotating sandwich complex. In a rotating magnetic field, the rotational frequency of a sandwich complex scales with the number of attached magnetic beads, which scales with the concentration of the protein present in solution. This paper represents the first instance of the detection of a protein using LAM. PMID:21805996

  20. 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.

  1. Elevated BMI and Male Sex Are Associated with Greater Underreporting of Caloric Intake as Assessed by Doubly Labeled Water12

    PubMed Central

    Stice, Eric; Palmrose, Christina A; Burger, Kyle S

    2015-01-01

    Background: Inaccuracies in energy intake (EI) measurement hinder identification of risk factors that predict weight gain and evaluation of obesity prevention and treatment interventions. Research has used objective measures of EI to identify underreporting correlates, producing mixed results, suggesting the need to examine novel potential correlates. Objective: With the use of an objective measure of EI from doubly labeled water (DLW) this report examined multiple potential underreporting correlates. Methods: Adolescents from 2 studies (study 1, n = 91; mean age: 18.4 ± 0.58 y; 100% female; study 2, n = 162; mean age: 15.2 ± 1.99 y; 82 female adolescents; 80 male adolescents) completed a DLW assessment of EI, a food-frequency questionnaire, and measures of perceived pressure for thinness, thin-ideal internalization, body dissatisfaction, dieting, food-cue reactivity, eating disorder symptoms, socioeconomic status, and neural response to food; BMI (in kg/m2) was measured over a 2-y follow-up. Results: Elevated BMI correlated with underreported EI in study 1 (r = 0.26, P < 0.05) and study 2 (r = 0.20, P = 0.01), as did male sex in study 2 (r = 0.24, P < 0.01); the other survey measures did not. Underreporting correlated negatively (r = −0.29; uncorr P < 0.001) with responsivity of brain regions implicated in motor control to palatable food receipt and positively (r = 0.31; uncorr P < 0.001) with responsivity of a region implicated in taste processing to cues signaling impending milkshake receipt. Underreporting did not predict future change in BMI in either study. Conclusions: Findings document marked underreporting and replicate evidence that BMI correlates positively with underreporting and extends this literature by revealing that several novel factors were unrelated to underreporting and further that neural responsivity to food correlated with underreporting, suggesting that adolescents who showed reduced responsivity in a motor control region to food

  2. 14C-most-probable-number method for enumeration of active heterotrophic microorganisms in natural waters.

    PubMed Central

    Lehmicke, L G; Williams, R T; Crawford, R L

    1979-01-01

    A most-probable-number method using 14C-labeled substrates is described for the enumeration of aquatic populations of heterotrophic microorganisms. Natural populations of microorganisms are inoculated into dilution replicates prepared from the natural water from which the organisms originated. The natural water is supplemented with a 14C-labeled compound added so as to approximate a true environmental concentration. 14CO2 evolved by individual replicates is trapped in NaOH and counted by liquid scintillation techniques for use in scoring replicates as positive or negative. Positives (14CO2 evolution) are easily distinguished from negatives (no 14CO2 evolution). The results from a variety of environments using the 14CO2 procedure agreed well with previously described methods, in most instances. The 14C-most-probable-number method described here reduces handling procedures over previously described most-probable-number procedures using 14C-labeled substrates. It also appears to have advantages over other enumeration methods in its attempt to approximate natural conditions more closely. PMID:120133

  3. 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.

  4. 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.

  5. 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.

  6. Method for single illumination source combined optical coherence tomography and fluorescence imaging of fluorescently labeled ocular structures in transgenic mice.

    PubMed

    McNabb, Ryan P; Blanco, Tomas; Bomze, Howard M; Tseng, Henry C; Saban, Daniel R; Izatt, Joseph A; Kuo, Anthony N

    2016-10-01

    In vivo imaging permits longitudinal study of ocular disease processes in the same animal over time. Two different in vivo optical imaging modalities - optical coherence tomography (OCT) and fluorescence - provide important structural and cellular data respectively about disease processes. In this Methods in Eye Research article, we describe and demonstrate the combination of these two modalities producing a truly simultaneous OCT and fluorescence imaging system for imaging of fluorescently labeled animal models. This system uses only a single light source to illuminate both modalities, and both share the same field of view. This allows simultaneous acquisition of OCT and fluorescence images, and the benefits of both techniques are realized without incurring increased costs in variability, light exposure, time, and post-processing effort as would occur when the modalities are used separately. We then utilized this system to demonstrate multi-modal imaging in a progression of samples exhibiting both fluorescence and OCT scattering beginning with resolution targets, ex vivo thy1-YFP labeled neurons in mouse eyes, and finally an in vivo longitudinal time course of GFP labeled myeloid cells in a mouse model of ocular allergy. PMID:27519152

  7. 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.

  8. A label-free turn-on fluorescence probe for rapidly distinguishing cysteine over glutathione in water solution.

    PubMed

    Yan, Liqiang; Kong, Zhineng; Shen, Wei; Du, Wenqi; Zhou, Yan; Qi, Zhengjian

    2016-05-01

    A novel label-free fluorescent chemodosimeter (C1) was synthesized, based on coumarin and N-(4-aminobenzoyl)-β-alanine, for the selective detection of cysteine (Cys) over glutathione (GSH), which involved a click reaction of Cys to CN of a Schiff base. The probe C1 featured a fast response (about 3 min), emission in the visible region, and high selectivity. Addition of Cys in HEPES-NaOH solution (pH 7.4) to C1 in water resulted in the appearance of a new emission peak at 445 nm, in company with remarkable enhancement of fluorescence intensity, while other amino acids did not induce any significant fluorescence change. Meanwhile, the addition reaction of Cys to C1 elicited 90.8-fold fluorescence intensity enhancement, which resulted in a change of emission color from orange to blue. PMID:26869082

  9. A label-free turn-on fluorescence probe for rapidly distinguishing cysteine over glutathione in water solution.

    PubMed

    Yan, Liqiang; Kong, Zhineng; Shen, Wei; Du, Wenqi; Zhou, Yan; Qi, Zhengjian

    2016-05-01

    A novel label-free fluorescent chemodosimeter (C1) was synthesized, based on coumarin and N-(4-aminobenzoyl)-β-alanine, for the selective detection of cysteine (Cys) over glutathione (GSH), which involved a click reaction of Cys to CN of a Schiff base. The probe C1 featured a fast response (about 3 min), emission in the visible region, and high selectivity. Addition of Cys in HEPES-NaOH solution (pH 7.4) to C1 in water resulted in the appearance of a new emission peak at 445 nm, in company with remarkable enhancement of fluorescence intensity, while other amino acids did not induce any significant fluorescence change. Meanwhile, the addition reaction of Cys to C1 elicited 90.8-fold fluorescence intensity enhancement, which resulted in a change of emission color from orange to blue.

  10. Confocal laser scanning microscopic photoconversion: a new method to stabilize fluorescently labeled cellular elements for electron microscopic analysis

    PubMed Central

    Colello, Raymond J.; Tozer, Jordan; Henderson, Scott C.

    2012-01-01

    Photoconversion, the method by which a fluorescent dye is transformed into a stable, osmiophilic product that can be visualized by electron microscopy, is the most widely used method to enable the ultrastructural analysis of fluorescently-labeled cellular structures. Nevertheless, the conventional method of photoconversion using widefield fluorescence microscopy requires long reaction times and results in low resolution cell targeting. Accordingly, we have developed a photoconversion method that ameliorates these limitations by adapting confocal laser scanning microscopy to the procedure. We have found that this method greatly reduces photoconversion times as compared to conventional wide field microscopy. Moreover, region of interest scanning capabilities of a confocal microscope facilitate the targeting of the photoconversion process to individual cellular or subcellular elements within a fluorescent field. This reduces the area of the cell exposed to light energy, thereby reducing the ultrastructural damage common to this process when widefield microscopes are employed. PMID:23042499

  11. 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.

  12. 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. PMID:25636019

  13. 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

  14. 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

  15. METHODS FOR DETERMINING RECREATIONAL WATER QUALITY

    EPA Science Inventory

    The goal of the clean water act of 1972 was to restore and maintain physical, chemical & biological quality of waters in the U.S. Although great progress has been made in cleaning up lakes, rivers and coastal waters many still do not meet water quality standards. Most beaches ha...

  16. Localization of cannabinoid CB1 receptor mRNA using ribonucleotide probes: methods for double- and single-label in situ hybridization.

    PubMed

    Hohmann, Andrea G

    2006-01-01

    This chapter presents a reliable, detailed method for performing double-label in situ hybridization (ISH) that has been validated for use in studies identifying the co-localization of cannabinoid CB1 receptor mRNA with other distinct species of mRNAs. This method permits simultaneous detection of two different species of mRNA within the same tissue section. Double-label ISH may be accomplished by hybridizing tissue sections with a combination of radiolabeled and digoxigenin-labeled RNA probes that are complementary to their target mRNAs. Single-label ISH may be accomplished by following the procedures described for use with radioisotopic probes (here [35S]-labeled) only. Silver grains derived from conventional emulsion autoradiography are used to detect the radiolabeled cRNA probe. An alkaline phosphatase-dependent chromogen reaction product is used to detect the nonisotopic (here, digoxigenin-labeled) cRNA probe. Necessary controls that are required to document the specificity of the labeling of the digoxigenin and radiolabeled probes are described. The methods detailed herein may be employed to detect even low levels of a target mRNA. These methods may be utilized to study co-localization and coregulation of expression of a particular gene within identified neurons in multiple systems.

  17. Method of extracting uranium from sea water

    SciTech Connect

    Paschke, M.; Wagener, K.; Wald, M.

    1981-04-21

    A method is described for producing a matrix of micro-organisms for extracting and gaining uranium from sea water, according to which the matrix is made of micro-organisms occurring in nature that are exposed to x-ray irradiations. These predetermined micro-organisms accordingly form colonies or threads and are filtered easily and cultivated. These microorganisms have a ratio of surface to volume that is relatively high. Subsequently, the irradiated micro-organisms are placed on a nutrient medium rich in uranium and are left there until colonies are formed. Thereupon, the surviving colonies of mutants are inoculated in a nutrient solution, and are then cultivated to produce the matrix.

  18. Synthesis of [sup 14]C-labeled C[sub 60], its suspension in water, and its uptake by human keratinocytes

    SciTech Connect

    Scrivens, W.A.; Tour, J.M.; Creek, K.E.; Pirisi, L. )

    1994-05-18

    We describe how, in order for us to determine whether unfunctionalized parent C[sub 60] had any biological activity, we synthesized [sup 14]C-labeled C[sub 60], developed a method to produce a fine aqueous suspension of the labeled material, and then monitored the uptake of the labeled C[sub 60] by human cells. The C[sub 60] became rapidly cell-associated, though it did not affect the proliferation of human keratinocytes or human fibroblasts, indicating that the rapid accumulation of C[sub 60] in human cells does not result in acute toxicity. 17 refs., 1 fig.

  19. 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.

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

    PubMed

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

    2016-03-01

    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.

  1. 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.

  2. Stable isotope labeling method for the investigation of protein haptenation by electrophilic skin sensitizers.

    PubMed

    Parkinson, Erika; Boyd, Pete; Aleksic, Maja; Cubberley, Richard; O'Connor, David; Skipp, Paul

    2014-11-01

    The risk of contact sensitization is a major consideration in the development of new formulations for personal care products. However, developing a mechanistic approach for non-animal risk assessment requires further understanding of haptenation of skin proteins by sensitizing chemicals, which is the molecular initiating event causative of skin sensitization. The non-stoichiometric nature of protein haptenation results in relatively low levels of modification, often of low abundant proteins, presenting a major challenge for their assignment in complex biological matrices such as skin. Instrumental advances over the last few years have led to a considerable increase in sensitivity of mass spectrometry (MS) techniques. We have combined these advancements with a novel dual-labeling/LC-MS(E) approach to provide an in-depth direct comparison of human serum albumin (HSA), 2,4-dinitro-1-chlorobenzene (DNCB), 5-chloro-2-methyl-4-isothiazolin-3-one (MCI), trans-cinnamaldehyde, and 6-methyl coumarin. These data have revealed novel insights into the differences in protein haptenation between sensitizers with different reaction mechanisms and sensitizing potency; the extreme sensitizers DNCB and MCI were shown to modify a greater number of nucleophilic sites than the moderate sensitizer cinnamaldehyde; and the weak/non-sensitizer 6-methyl coumarin was restricted to only a single nucleophilic residue within HSA. The evaluation of this dual labeling/LC-MS(E) approach using HSA as a model protein has also demonstrated that this strategy could be applied to studying global haptenation in complex mixtures of skin-related proteins by different chemicals.

  3. 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

  4. 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

  5. Label-Free Fluorescence Assay of S1 Nuclease and Hydroxyl Radicals Based on Water-Soluble Conjugated Polymers and WS₂ Nanosheets.

    PubMed

    Li, Junting; Zhao, Qi; Tang, Yanli

    2016-06-13

    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.

  6. Methods for virus recovery in water

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  7. Detection of follicular apoptosis in water buffalo (Bubalus bubalis) ovary by histology and nick end labelling technique.

    PubMed

    Sreejalekshmi, P; Raghavendra, B S; Subramani, T Siva; Murthy, V Chandrashekara; Jamuna, K V; Prasad, R V; Ravindra, J P; Selvaraju, S

    2011-02-01

    The objective of this experiment was to assess the features and extent of follicular apoptosis in the water buffalo (Bubalus bubalis) ovary using classical histology and nick end labelling technique. Ovaries (n=40) procured from the slaughterhouse were used for the study. The sections (5 μm) were used for detection of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) and classical histology (H&E). Those follicles showing ≥ 5% TUNEL positivity (TUNEL assay) and pyknotic nuclei (histology) in granulosa cells were classified as atretic. Based on histology, the atretic primary and secondary follicles (%) were 93.82 and 95.62 respectively. The histology study reveals that the rates (%) of atresia in <1, 1-3, 3-5 mm and >5 mm were 36.90, 40.50, 62.84 and 74.5 respectively. Further the atretic tertiary follicles (%) were significantly lower than the primary and secondary classes of follicles. TUNEL assay reveals that the atretic rate (%) of tertiary follicles in <1, 1-3, 3-5 and ≥ 5 mm class follicles were 50.88, 53.84, 81.81 and 36.36 respectively. The percentage of atresia in >5 mm diameter follicles were significantly lower in TUNEL than histology. Percentages of granulosa and thecal cells positive for atresia by TUNEL were 30.7 ± 0.53 and 13.82 ± 0.18 respectively per follicle. The initial structural changes in atretic follicles were seen primarily in the granulosa cells. In severely atretic follicles TUNEL positive granulosa cells along with theca cells have to be considered in assessing the rate and extent of atresia.

  8. 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

  9. 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

  10. Remote loading of diclofenac, insulin and fluorescein isothiocyanate labeled insulin into liposomes by pH and acetate gradient methods.

    PubMed

    Hwang, S H; Maitani, Y; Qi, X R; Takayama, K; Nagai, T

    1999-03-01

    Remote loading of the model drugs diclofenac, insulin and fluorescein isothiocyanate labeled insulin (FITC-insulin) into liposomes by formation of transmembrane gradients were examined. A trapping efficiency of almost 100% was obtained for liposomal diclofenac, by the calcium acetate gradient method, whereas liposomes prepared by the conventional reverse-phase evaporation vesicle method had 1-8% trapping efficiencies. Soybean-derived sterol was a better stabilizer of the dipalmitoylphosphatidylcholine bilayer membrane than cholesterol, as shown from trapping efficiencies and drug release. The pH gradient method resulted in a 5-50% of FITC-insulin liposomal trapping efficiency, while insulin could not be loaded by this method. Liposomes released calcein in response to insulin, showing insulin interacts with the liposomal membrane in the presence of a transmembrane gradient. The present work has demonstrated a remote loading method for weak acids such as diclofenac into liposomes by the acetate gradient method. From the result of remote loading of FITC-insulin into liposomes by the pH gradient method, this method may be available for the preparation of liposomal peptides.

  11. 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.

  12. 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.

  13. 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

  14. Synchrotron X-ray footprinting as a method to visualize water in proteins.

    PubMed

    Gupta, Sayan; Feng, Jun; Chan, Leanne Jade G; Petzold, Christopher J; Ralston, Corie Y

    2016-09-01

    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

  15. Synchrotron X-ray footprinting as a method to visualize water in proteins.

    PubMed

    Gupta, Sayan; Feng, Jun; Chan, Leanne Jade G; Petzold, Christopher J; Ralston, Corie Y

    2016-09-01

    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.

  16. Surface water quality assessment by environmetric methods.

    PubMed

    Boyacioglu, Hülya; Boyacioglu, Hayal

    2007-08-01

    This environmetric study deals with the interpretation of river water monitoring data from the basin of the Buyuk Menderes River and its tributaries in Turkey. Eleven variables were measured to estimate water quality at 17 sampling sites. Factor analysis was applied to explain the correlations between the observations in terms of underlying factors. Results revealed that, water quality was strongly affected from agricultural uses. Cluster analysis was used to classify stations with similar properties and results distinguished three groups of stations. Water quality at downstream of the river was quite different from the other part. It is recommended to involve the environmetric data treatment as a substantial procedure in assessment of water quality data.

  17. 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

  18. An innovative, quick and convenient labeling method for the investigation of pharmacological behavior and the metabolism of poly(DL-lactide-co-glycolide) nanospheres.

    PubMed

    Stevanović, Magdalena; Maksin, Tatjana; Petković, Jana; Filipic, Metka; Uskoković, Dragan

    2009-08-19

    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 (99m)Tc 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 (99m)Tc-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. PMID:19636100

  19. International experiences in assessing vitamin A status and applying the vitamin A-labeled isotope dilution method.

    PubMed

    Lopez-Teros, Veronica; Chileshe, Justin; Idohou-Dossou, Nicole; Fajarwati, Tetra; Medoua Nama, Gabriel; Newton, Sam; Vinod Kumar, Malavika; Wang, Zhixu; Wasantwisut, Emorn; Hunt, Janet R

    2014-01-01

    Inadequate vitamin A (VA) nutrition continues to be a major problem worldwide, and many interventions being implemented to improve VA status in various populations need to be evaluated. The interpretation of results after an intervention depends greatly on the method selected to assess VA status. To evaluate the effect of an intervention on VA status, researchers in Cameroon, India, Indonesia, Mexico, Senegal and Zambia have used serum retinol as an indicator, and have not always found improvement in response to supplementation. One problem is that homeostatic control of serum retinol may mask positive effects of treatment in that changes in concentration are observed only when status is either moderately to severely depleted or excessive. Because VA is stored mainly in the liver, measurements of hepatic VA stores are the “gold standard” for assessing VA status. Dose response tests such as the relative dose response (RDR) and the modified relative dose response (MRDR), allow a qualitative assessment of VA liver stores. On the other hand, the use of the vitamin A-labeled isotope dilution (VALID) technique, (using 13C or 2H-labeled retinyl acetate) serves as an indirect method to quantitatively estimate total body and liver VA stores. Countries including Cameroon, China, Ghana, Mexico, Thailand and Zambia are now applying the VALID method to sensitively assess changes in VA status during interventions, or to estimate a population’s dietary requirement for VA. Transition to the use of more sensitive biochemical indicators of VA status such as the VALID technique is needed to effectively assess interventions in populations where mild to moderate VA deficiency is more prevalent than severe deficiency. PMID:25537105

  20. 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.

  1. 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.

  2. Water turbine system and method of operation

    DOEpatents

    Costin, Daniel P.

    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.

  3. Water turbine system and method of operation

    DOEpatents

    Costin, Daniel P.

    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.

  4. 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...

  5. 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

  6. Numerical evaluation of subsoil diffusion of (15) N labelled denitrification products during employment of the (15) N gas flux method in the field

    NASA Astrophysics Data System (ADS)

    Well, Reinhard; Buchen, Caroline; Lewicka-Szczebak, Dominika; Ruoss, Nicolas

    2016-04-01

    Common methods for measuring soil denitrification in situ include monitoring the accumulation of 15N labelled N2 and N2O evolved from 15N labelled soil nitrate pool in soil surface chambers. Gas diffusion is considered to be the main accumulation process. Because accumulation of the gases decreases concentration gradients between soil and chamber over time, gas production rates are underestimated if calculated from chamber concentrations. Moreover, concentration gradients to the non-labelled subsoil exist, inevitably causing downward diffusion of 15N labelled denitrification products. A numerical model for simulating gas diffusion in soil was used in order to determine the significance of this source of error. Results show that subsoil diffusion of 15N labelled N2 and N2O - and thus potential underestimation of denitrification derived from chamber fluxes - increases with cover closure time as well as with increasing diffusivity. Simulations based on the range of typical gas diffusivities of unsaturated soils show that the fraction of subsoil diffusion after chamber closure for 1 hour is always significant with values up to >30 % of total production of 15N labelled N2 and N2O. Field experiments for measuring denitrification with the 15N gas flux method were conducted. The ability of the model to predict the time pattern of gas accumulation was evaluated by comparing measured 15N2 concentrations and simulated values.

  7. 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

  8. Method Development and Monitoring of Cyanotoxins in Water

    EPA Science Inventory

    This presentation describes method development of two ambient water LC/MS/MS methods for microcystins, cylindrospermopsin and anatoxin-a. Ruggedness of the methods will be demonstrated by evaluation of quality control samples derived from various water bodies across the country.

  9. A facile method for expression and purification of (15)N isotope-labeled human Alzheimer's β-amyloid peptides from E. coli for NMR-based structural analysis.

    PubMed

    Sharma, Sudhir C; Armand, Tara; Ball, K Aurelia; Chen, Anna; Pelton, Jeffrey G; Wemmer, David E; Head-Gordon, Teresa

    2015-12-01

    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 (15)N isotope-labeled Aβ42 for structural studies by NMR. The protocol involves utilization of an auto induction system with (15)N isotope labeled medium, for high-level expression of Aβ42 as a fusion with IFABP. After the over-expression of the (15)N isotope-labeled IFABP-Aβ42 fusion protein in the inclusion bodies, pure (15)N 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 (15)N isotope-labeled Aβ42 peptide. Mass spectrometry and (1)H-(15)N 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 (15)N and (13)C in Aβ42 peptide as well as its other variants including any Aβ42 peptide mutants.

  10. 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

  11. 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.

  12. 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

  13. INCLUSION OF 13C12-LABELLED MONO-, DI-, AND TRI-CHLORINATED DIBENZO-P-DIOXIN AND DIBENZOFURAN STANDARDS IN U.S. EPA METHODS 0023A/8290

    EPA Science Inventory

    13C12-labeled mono-, di-, and tri-chlorinated dibenzo-p-dioxin (CDD) and -chlorinated dibenzofuran (CDF) standards have been tested for their applicability to standard EPA sampling and analytical Methods 0023A/8290. These methods target for analysis only the tetra- through octa-C...

  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. 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.

  16. Potential of 99mTc-LDLs labeled by two different methods for scintigraphic detection of experimental atherosclerosis in rabbits.

    PubMed

    Atsma, D E; Feitsma, R I; Camps, J; van't Hooft, F M; van der Wall, E E; Nieuwenhuizen, W; Pauwels, E K

    1993-01-01

    In this study we evaluated two different 99mTc-labeling techniques to produce 99mTc-low density lipoprotein (99mTc-LDL) suitable for the scintigraphic delineation of experimental atherosclerotic lesions. The two methods are 1) a procedure that uses stannous chloride and sodium borohydride (borohydride method) and 2) a procedure that uses sodium dithionite as a reducing agent and that has been successfully applied in previous scintigraphic atherosclerosis detection (dithionite method). 99mTc-LDL produced by either method was injected into New Zealand White rabbits with diet-induced atherosclerotic plaques and in control rabbits. Scintigraphic images were taken 10 minutes (t = 0) and 1, 4, 8, 16, and 24 hours after injection. Clearance of plasma radioactivity was also studied. Stability of the 99mTc-LDL complex in the circulation was examined by size exclusion chromatography of plasma samples. After scintigraphy, the animals were killed, and the biodistribution of radioactivity was determined. The thoracic and abdominal aortas appeared in scintigraphic images to accumulate 99mTc over their entire length with either 99mTc-LDL preparation. The sparse imaging of focal atherosclerosis was found to be due to the fact that the aortas were covered with confluent atherosclerotic lesions. Scintigraphic image analysis showed that 24 hours after injection, the accumulated radioactivity in the abdominal aorta of the atherosclerotic rabbits was 57% and 54%, respectively, of the accumulated radioactivity in the abdominal aorta at t = 0 when the borohydride versus the dithionite method was used. In the control animals this value was 25% for the dithionite method, whereas in the borohydride method the aortas could not be detected in the images at t = 24 hours.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8422342

  17. A novel multiplex RT-qPCR method based on dual-labelled probes suitable for typing all known genotypes of viral haemorrhagic septicaemia virus.

    PubMed

    Vázquez, D; López-Vázquez, C; Skall, H F; Mikkelsen, S S; Olesen, N J; Dopazo, C P

    2016-04-01

    Viral haemorrhagic septicaemia (VHS) is a notifiable fish disease, whose causative agent is a rhabdovirus isolated from a wide range of fish species, not only in fresh but also in marine and brackish waters. Phylogenetic studies have identified four major genotypes, with a strong geographical relationship. In this study, we have designed and validated a new procedure--named binary multiplex RT-qPCR (bmRT-qPCR)--for simultaneous detection and typing of all four genotypes of VHSV by real-time RT-PCR based on dual-labelled probes and composed by two multiplex systems designed for European and American/Asiatic isolates, respectively, using a combination of three different fluorophores. The specificity of the procedure was assessed by including a panel of 81 VHSV isolates covering all known genotypes and subtypes of the virus, and tissue material from experimentally infected rainbow trout, resulting in a correct detection and typing of all strains. The analytical sensitivity was evaluated in a comparative assay with titration in cell culture, observing that both methods provided similar limits of detection. The proposed method can be a powerful tool for epidemiological analysis of VHSV by genotyping unknown samples within a few hours.

  18. 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

    2016-08-01

    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%. PMID:27403652

  19. 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

    2016-08-01

    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%.

  20. 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.

  1. 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.

  2. How to save water by choice of irrigation application method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. Review of methods for assessing nonpoint-source contaminated ground-water discharge to surface water

    SciTech Connect

    Not Available

    1991-04-01

    The document provides an overview of selected methods that have been used for assessing nonpoint source contaminated ground water discharge to surface water. EPA undertook the project in response to the growing awareness that contaminated ground water discharge is a significant source of nonpoint source contaminant loading to surface water in many parts of the country.

  4. Study of carbohydrate structure and reactivity by modern NMR methods and isotopic labeling

    SciTech Connect

    Snyder, J.R.

    1987-01-01

    Chemical methods are described for preparing unenriched and (1-/sup 13/C)-enriched 5-deoxy- and 5-O-methylpentoses in the D or L configuration. The /sup 1/H and /sup 13/C NMR spectra of these compounds have been interpreted and the carbon spectra assigned with the aid of 2D /sup 13/C-/sup 1/H chemical-shift correlation spectroscopy. The tautomeric composition in /sup 2/H/sub 2/O has been quantitated with the aid of (1-/sup 13/C)-enriched derivatives. The branched-chain pentose, DL-apiose has been synthesized in good yield by a new and simple chemical method that can be adapted to prepare (1-/sup 13/C)-, (2-/sup 13/C)-, (1-/sup 2/H)- and/or (2-/sup 2/H)-enriched derivatives. The solution composition of D-idose in D/sup 2/O has been examined by /sup 13/C NMR spectroscopy using (/sup 13/C)-enriched compounds. In addition to two furanoses and two pyranoses, aldehyde and hydrate forms have been detected and quantified. The solution composition of D-talose has been investigated by /sup 13/C NMR spectroscopy using (1-/sup 13/C)talose. The tautomeric composition has been determined at 28/sup 0/, and the results show equivalent amounts of the acyclic aldehyde and hydrate. Several structurally modified furanose sugars were synthesized to assess the extent the Thorpe-Ingold effect promotes rings formation and enhances rates of ring-closure.

  5. 99m tc labeled liposomes

    SciTech Connect

    Phillips, W.T.; Klipper, R.W.; Timmons, J.H.; Rudolph, A.S.

    1992-10-27

    This patent describes a method of preparing stable gamma-emitting radionuclide-labeled alkyleneamine oxime, the incubating being for a period of time sufficient to form labeled liposome-encapsulated protein.

  6. 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.

  7. 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

  8. 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.

  9. 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

  10. General, Label-Free Method for Determining K(d) and Ligand Concentration Simultaneously.

    PubMed

    Jalali-Yazdi, Farzad; Takahashi, Terry T; Roberts, Richard W

    2015-12-01

    Some of the most commonly used affinity reagents (e.g., antibodies) are often developed and used in conditions where their input concentrations ([L]0) and affinities (K(d)) are not known. Here, we have developed a general approach to determine both [L]0 and K(d) values simultaneously for affinity reagents (small molecules, proteins, and antibodies). To do this, we perform quantitative equilibrium exclusion immunoassays with two different concentrations of target and fit the data simultaneously to determine K(d) and [L]0. The results give accurate and reproducible measures of both values compared to established methods. By performing detailed error analysis, we demonstrate that our fitting gives unique solutions and indicates where K(d) and [L]0 measures are reliable. Furthermore, we found that a divalent model of antibody binding gives accurate K(d) and [L]0 values in both the forward (antibody immobilized) and the reverse (target immobilized) assays-addressing the long-term problem of obtaining quantitative data from reverse assays.

  11. 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.

  12. Use of Dimedone-Based Chemical Probes for Sulfenic Acid Detection: Methods to Visualize and Identify Labeled Proteins

    PubMed Central

    Nelson, Kimberly J.; Klomsiri, Chananat; Codreanu, Simona G.; Soito, Laura; Liebler, Daniel C.; Rogers, LeAnn C.; Daniel, Larry W.; Poole, Leslie B.

    2013-01-01

    Reversible thiol modification is a major component of the modulation of cell-signaling pathways by reactive oxygen species. Hydrogen peroxide, peroxynitrite, or lipid hydroperoxides are all able to oxidize cysteines to form cysteine sulfenic acids; this reactive intermediate can be directly reduced to thiol by cellular reductants such as thioredoxin or further participate in disulfide bond formation with glutathione or cysteine residues in the same or another protein. To identify the direct protein targets of cysteine modification and the conditions under which they are oxidized, a series of dimedone-based reagents linked to affinity or fluorescent tags have been developed that specifically alkylate and trap cysteine sulfenic acids. In this chapter, we provide detailed methods using one of our biotin-tagged reagents, DCP-Bio1, to identify and monitor proteins that are oxidized in vitro and in vivo. Using streptavidin-linked agarose beads, this biotin-linked reagent can be used to affinity capture labeled proteins. Stringent washing of the beads prior to elution minimizes the contamination of the enriched material with unlabeled proteins through coimmunoprecipitation or nonspecific binding. In particular, we suggest including DTT in one of the washes to remove proteins covalently linked to biotinylated proteins through a disulfide bond, except in cases where these linked proteins are of interest. We also provide methods for targeted approaches monitoring cysteine oxidation in individual proteins, global approaches to follow total cysteine oxidation in the cell, and guidelines for proteomic analyses to identify novel proteins with redox sensitive cysteines. PMID:20513473

  13. Comparison of Protein N-Homocysteinylation in Rat Plasma under Elevated Homocysteine Using a Specific Chemical Labeling Method.

    PubMed

    Zang, Tianzhu; Pottenplackel, Ligi Paul; Handy, Diane E; Loscalzo, Joseph; Dai, Shujia; Deth, Richard C; Zhou, Zhaohui Sunny; Ma, Jisheng

    2016-01-01

    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

  14. 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.

  15. 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.

  16. 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.

  17. 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)

  18. 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

  19. 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.

  20. Label fusion strategy selection.

    PubMed

    Robitaille, Nicolas; Duchesne, Simon

    2012-01-01

    Label fusion is used in medical image segmentation to combine several different labels of the same entity into a single discrete label, potentially more accurate, with respect to the exact, sought segmentation, than the best input element. Using simulated data, we compared three existing label fusion techniques-STAPLE, Voting, and Shape-Based Averaging (SBA)-and observed that none could be considered superior depending on the dissimilarity between the input elements. We thus developed an empirical, hybrid technique called SVS, which selects the most appropriate technique to apply based on this dissimilarity. We evaluated the label fusion strategies on two- and three-dimensional simulated data and showed that SVS is superior to any of the three existing methods examined. On real data, we used SVS to perform fusions of 10 segmentations of the hippocampus and amygdala in 78 subjects from the ICBM dataset. SVS selected SBA in almost all cases, which was the most appropriate method overall. PMID:22518113

  1. Nutrition Labeling

    NASA Astrophysics Data System (ADS)

    Metzger, Lloyd E.

    Nutrition labeling regulations differ in countries around the world. The focus of this chapter is on nutrition labeling regulations in the USA, as specified by the Food and Drug Administration (FDA) and the Food Safety and Inspection Service (FSIS) of the United States Department of Agriculture (USDA). A major reason for analyzing the chemical components of foods in the USA is nutrition labeling regulations. Nutrition label information is not only legally required in many countries, but also is of increasing importance to consumers as they focus more on health and wellness.

  2. Methods for the determination of organic compounds in drinking water, Supplement 1

    SciTech Connect

    Not Available

    1990-07-01

    Nine analytical methods covering 54 organic contaminants which may be present in drinking water or drinking water sources are described in detail. Seven of these methods cover compounds designated for regulation under the Safe Drinking Water Act Amendments of 1986. Regulations for this group are in the proposal stages with promulgation scheduled for June 1992. The other two methods are for chlorination disinfection byproducts and may be regulated as part of EPA's disinfectants and disinfectant byproducts rule scheduled for proposal early in 1992. Most of the analytes may be classified as non-volatile and three of the methods entail separations by high performance liquid chromatography. The remainder employ capillary column gas chromatography. One of these requires detection of a potentially very toxic contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin, at the low parts per trillion level. Labeled isotopes of this analyte are employed as tracers and high resolution mass spectrometry is required for detection and unambiguous identification. Three of the methods herein offer new and simplified liquid-solid extraction procedures, a trend which is likely to become even more pronounced in the future.

  3. 40 CFR 600.115-08 - Criteria for determining the fuel economy label calculation method for 2011 and later model year...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... economy label calculation method for 2011 and later model year vehicles. 600.115-08 Section 600.115-08 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) ENERGY POLICY FUEL ECONOMY AND CARBON-RELATED EXHAUST EMISSIONS OF MOTOR VEHICLES Fuel Economy and Carbon-Related Exhaust Emission...

  4. 40 CFR 600.115-08 - Criteria for determining the fuel economy label calculation method for 2011 and later model year...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... economy label calculation method for 2011 and later model year vehicles. 600.115-08 Section 600.115-08 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) ENERGY POLICY FUEL ECONOMY AND CARBON-RELATED EXHAUST EMISSIONS OF MOTOR VEHICLES Fuel Economy Regulations for 1978 and Later Model...

  5. Novel tracer method to measure isotopic labeled gas-phase nitrous acid (HO15NO) in biogeochemical studies.

    PubMed

    Wu, Dianming; Kampf, Christopher J; Pöschl, Ulrich; Oswald, Robert; Cui, Junfang; Ermel, Michael; Hu, Chunsheng; Trebs, Ivonne; Sörgel, Matthias

    2014-07-15

    Gaseous nitrous acid (HONO), the protonated form of nitrite, contributes up to ∼60% to the primary formation of hydroxyl radical (OH), which is a key oxidant in the degradation of most air pollutants. Field measurements and modeling studies indicate a large unknown source of HONO during daytime. Here, we developed a new tracer method based on gas-phase stripping-derivatization coupled to liquid chromatography-mass spectrometry (LC-MS) to measure the 15N relative exceedance, ψ(15N), of HONO in the gas-phase. Gaseous HONO is quantitatively collected and transferred to an azo dye, purified by solid phase extraction (SPE), and analyzed using high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). In the optimal working range of ψ(15N)=0.2-0.5, the relative standard deviation of ψ(15N) is <4%. The optimum pH and solvents for extraction by SPE and potential interferences are discussed. The method was applied to measure HO15NO emissions from soil in a dynamic chamber with and without spiking 15) labeled urea. The identification of HO15NO from soil with 15N urea addition confirmed biogenic emissions of HONO from soil. The method enables a new approach of studying the formation pathways of HONO and its role for atmospheric chemistry (e.g., ozone formation) and environmental tracer studies on the formation and conversion of gaseous HONO or aqueous NO2- as part of the biogeochemical nitrogen cycle, e.g., in the investigation of fertilization effects on soil HONO emissions and microbiological conversion of NO2- in the hydrosphere.

  6. A quantitative method to measure human platelet chemotaxis using indium-111-oxine-labeled gel-filtered platelets

    SciTech Connect

    Lowenhaupt, R.W.; Silberstein, E.B.; Sperling, M.I.; Mayfield, G.

    1982-12-01

    Human blood platelets have been shown to migrate directionally and specifically toward collagen in plasma in vitro. We have developed a new system to monitor this behavior using a linear 7-compartment chamber with /sup 111/In-oxine-labeled gel-filtered platelets. The compartments are separated by various Nuclepore and Millipore filter membranes. Radiolabeled platelets suspended in plasma are placed in the central compartment and the other compartments are filled with platelet-free plasma. When collagen is added to an end compartment, platelets migrate toward that end. The degree of this directed movement or chemotaxis can be measured by counting the radioactivity of the contents of each compartment and then comparing the counts from radiolabeled platelets that have moved to the end that holds the chemotactic inducer with those that have randomly migrated to the opposite end, containing only plasma. This assay system allows quantitative comparisons between the chemotaxis-inducing abilities of different substances and permits the study of soluble materials. Experiments to determine the optimal conditons for the procedure are reported, and the advantages of this new method for the investigation of platelet chemotaxis and the identification of chemotaxins are discussed.

  7. A quantitative method to measure human platelet chemotaxis using /sup 111/In-oxine-labeled gel-filtered platelets

    SciTech Connect

    Lowenhaupt, R.W.; Silberstein, E.B.; Sperling, M.I.; Mayfield, G.

    1982-12-01

    Human blood platelets have been shown to migrate directionally and specifically toward collagen in plasma in vitro. We have developed a new system to monitor this behavior using a linear 7-compartment chamber with /sup 111/In-oxine-labeled gel-filtered platelets. The compartments are separated by various Nuclepore and Millipore filter membranes. Radiolabeled platelets suspended in plasma are placed in the central compartment and the other compartments are filled with platelet-free plasma. When collagen is added to an end compartment, platelets migrate toward that end. The degree of this directed movement or chemotaxis can be measured by counting the radioactivity of the contents of each compartment and then comparing the counts from radiolabeled platelets that have moved to the end that holds the chemotactic inducer with those that have randomly migrated to the opposite end, containing only plasma. This assay system allows quantitative comparisons between the chemotaxis-inducing abilities of different substances and permits the study of soluble materials. Experiments to determine the optimal conditions for the procedure are reported, and the advantages of this new method for the investigation of platelet chemotaxis and the identification of chemotaxins are discussed.

  8. Examining the Interactions of the Splicing Factor MBNL1 with Target RNA Sequences via a Label-Free, Multiplex Method

    PubMed Central

    Yadav, Amrita R.; Mace, Charles R.; Miller, Benjamin L.

    2014-01-01

    The near-ubiquity of the involvement of RNA in crucial biological processes is accepted. It is important, therefore, to study and understand the biophysical principles that regulate the function of RNA and its interactions with other molecules (e.g., proteins and antibiotics). Methods enabling the high-throughput determination of RNA–protein binding kinetics and thermodynamics would greatly accelerate understanding of these interactions. To that end, we describe the development of a real-time biomolecular interaction analysis platform based on arrayed imaging reflectometry (AIR) for multiplex analysis of RNA–protein interactions. We demonstrate the use of aqueous AIR by measuring the binding kinetics between muscleblind-like 1 (MBNL1), a splicing regulator protein that plays a pivotal role in the Myotonic Dystrophies and Huntington's Disease, and several of its RNA targets simultaneously on a microarrayed chip. Using this approach, we observe that the kinetics of MBNL1 binding isolated CUG and repeat CUG RNA sequences (as models for “normal” and “pathogenic” RNA, respectively) are different even though their steady state binding constants are similar. The ability to compare binding kinetics between RNA sequences rapidly and easily may provide insight into the molecular basis of MBNL1-RNA binding, and more generally suggests that AIR can be a powerful tool to enable the label-free, real-time analysis of biomolecular interactions in a high throughput format. PMID:24377303

  9. Compositions and methods for removing arsenic in water

    DOEpatents

    Gadgil, Ashok Jagannth

    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.

  10. Validation of Cross-Sectional Time Series and Multivariate Adaptive Regression Splines Models for the Prediction of Energy Expenditure in Children and Adolescents Using Doubly Labeled Water12

    PubMed Central

    Butte, Nancy F.; Wong, William W.; Adolph, Anne L.; Puyau, Maurice R.; Vohra, Firoz A.; Zakeri, Issa F.

    2010-01-01

    Accurate, nonintrusive, and inexpensive techniques are needed to measure energy expenditure (EE) in free-living populations. Our primary aim in this study was to validate cross-sectional time series (CSTS) and multivariate adaptive regression splines (MARS) models based on observable participant characteristics, heart rate (HR), and accelerometer counts (AC) for prediction of minute-by-minute EE, and hence 24-h total EE (TEE), against a 7-d doubly labeled water (DLW) method in children and adolescents. Our secondary aim was to demonstrate the utility of CSTS and MARS to predict awake EE, sleep EE, and activity EE (AEE) from 7-d HR and AC records, because these shorter periods are not verifiable by DLW, which provides an estimate of the individual's mean TEE over a 7-d interval. CSTS and MARS models were validated in 60 normal-weight and overweight participants (ages 5–18 y). The Actiheart monitor was used to simultaneously measure HR and AC. For prediction of TEE, mean absolute errors were 10.7 ± 307 kcal/d and 18.7 ± 252 kcal/d for CSTS and MARS models, respectively, relative to DLW. Corresponding root mean square error values were 305 and 251 kcal/d for CSTS and MARS models, respectively. Bland-Altman plots indicated that the predicted values were in good agreement with the DLW-derived TEE values. Validation of CSTS and MARS models based on participant characteristics, HR monitoring, and accelerometry for the prediction of minute-by-minute EE, and hence 24-h TEE, against the DLW method indicated no systematic bias and acceptable limits of agreement for pediatric groups and individuals under free-living conditions. PMID:20573939

  11. Methods for tritium labeling

    DOEpatents

    Andres, Hendrik; Morimoto, Hiromi; Williams, Philip G.

    1993-01-01

    Reagents and processes for reductively introducing deuterium or tritium into organic molecules are described. The reagents are deuterium or tritium analogs of trialkyl boranes, borane or alkali metal aluminum hydrides. The process involves forming these reagents in situ from alkali metal tritides or deuterides.

  12. Estimation of Free-Living Energy Expenditure by Heart Rate and Movement Sensing: A Doubly-Labelled Water Study

    PubMed Central

    Brage, Søren; Westgate, Kate; Franks, Paul W.; Stegle, Oliver; Wright, Antony; Ekelund, Ulf; Wareham, Nicholas J.

    2015-01-01

    Background Accurate assessment of energy expenditure (EE) is important for the study of energy balance and metabolic disorders. Combined heart rate (HR) and acceleration (ACC) sensing may increase precision of physical activity EE (PAEE) which is the most variable component of total EE (TEE). Objective To evaluate estimates of EE using ACC and HR data with or without individual calibration against doubly-labelled water (DLW) estimates of EE. Design 23 women and 23 men (22–55 yrs, 48–104 kg, 8–46%body fat) underwent 45-min resting EE (REE) measurement and completed a 20-min treadmill test, an 8-min step test, and a 3-min walk test for individual calibration. ACC and HR were monitored and TEE measured over 14 days using DLW. Diet-induced thermogenesis (DIT) was calculated from food-frequency questionnaire. PAEE (TEE ÷ REE ÷ DIT) and TEE were compared to estimates from ACC and HR using bias, root mean square error (RMSE), and correlation statistics. Results Mean(SD) measured PAEE and TEE were 66(25) kJ·day-1·kg-1, and 12(2.6) MJ·day-1, respectively. Estimated PAEE from ACC was 54(15) kJ·day-1·kg-1 (p<0.001), with RMSE 24 kJ·day-1·kg-1 and correlation r = 0.52. PAEE estimated from HR and ACC+HR with treadmill calibration were 67(42) and 69(25) kJ·day-1·kg-1 (bias non-significant), with RMSE 34 and 20 kJ·day-1·kg-1 and correlations r = 0.58 and r = 0.67, respectively. Similar results were obtained with step-calibrated and walk-calibrated models, whereas non-calibrated models were less precise (RMSE: 37 and 24 kJ·day-1·kg-1, r = 0.40 and r = 0.55). TEE models also had high validity, with biases <5%, and correlations r = 0.71 (ACC), r = 0.66–0.76 (HR), and r = 0.76–0.83 (ACC+HR). Conclusions Both accelerometry and heart rate may be used to estimate EE in adult European men and women, with improved precision if combined and if heart rate is individually calibrated. PMID:26349056

  13. 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.

  14. 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.

  15. 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.

  16. Dosimetry of intravenously administered oxygen-15 labelled water in man: a model based on experimental human data from 21 subjects.

    PubMed

    Smith, T; Tong, C; Lammertsma, A A; Butler, K R; Schnorr, L; Watson, J D; Ramsay, S; Clark, J C; Jones, T

    1994-10-01

    Models based on uniform distribution of tracer in total body water underestimate the absorbed dose from H2(15)O because of the short half-life (2.04 min) of 15O, which leads to non-uniform distribution of absorbed dose and also complicates the direct measurement of organ retention curves. However, organ absorbed doses can be predicted by the present kinetic model based on the convolution technique. The measured time course of arterial H2(15)O concentration following intravenous administration represents the input function to organs. The impulse response of a given organ is its transit time function determined by blood flow and the partition of water between tissue and blood. Values of these two parameters were taken from the literature. Integrals of the arterial input function and organ transit time functions were used to derive integrals of organ retention functions (organ residence times). The latter were used with absorbed dose calculation software (MIRDOSE-2) to obtain estimates for 24 organs. From the mean values of organ absorbed doses, the effective dose equivalent (EDE) and effective dose (ED) were calculated. From measurements on 21 subjects, the average value for both EDE and ED was calculated to be 1.2 microSv.MBq-1 compared with a value of about 0.5 microSv.MBq-1 predicted by uniform water distribution models. Based on the human data, a method of approximating H2(15)O absorbed dose values from body surface area is described.

  17. 78 FR 78305 - Energy and Water Use Labeling for Consumer Products Under the Energy Policy and Conservation Act...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-26

    ... proceeding, DOE announced plans to develop a new test procedure. 74 FR 53640, 53641 (Oct. 20, 2009). \\2\\ 16... published on October 25, 2013 (78 FR 63823). These amendments will ensure the Rule's television labeling... televisions in 2011 (76 FR 1038 (Jan. 6, 2011)), no DOE test procedure existed for such products....

  18. 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)

  19. 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...

  20. 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.

  1. 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 ...

  2. Inulin determination for food labeling.

    PubMed

    Zuleta, A; Sambucetti, M E

    2001-10-01

    Inulin and oligofructose exhibit valuable nutritional and functional attributes, so they are used as supplements as soluble fiber or as macronutrient substitutes. As classic analytical methods for dietary fiber measurement are not effective, several specific methods have been proposed. These methods measure total fructans and are based on one or more enzymatic sample treatments and determination of released sugars. To determine inulin for labeling purposes, we developed an easy and rapid anion-exchange high-performance liquid chromatography (HPLC) method following water extraction of inulin. HPLC conditions included an Aminex HPX- 87C column (Bio-Rad), deionized water at 85 degrees C as the mobile phase and a refractive index detector. The tested foods included tailor-made food products containing known amounts of inulin and commercial products (cookies, milk, ice creams, cheese, and cereal bars). The average recovery was 97%, and the coefficient of variation ranged from 1.1 to 5% in the food matrixes. The obtained results showed that this method provides an easier, faster and cheaper alternative than previous techniques for determining inulin with enough accuracy and precision for routine labeling purposes by direct determination of inulin by HPLC with refractive index detection. PMID:11599989

  3. Synthesis of a Potent Aminopyridine-Based nNOS-Inhibitor by Two Recent No-Carrier-Added (18)F-Labelling Methods.

    PubMed

    Drerup, Christian; Ermert, Johannes; Coenen, Heinz H

    2016-01-01

    Nitric oxide (NO), an important multifunctional signaling molecule, is produced by three isoforms of NO-synthase (NOS) and has been associated with neurodegenerative disorders. Selective inhibitors of the subtypes iNOS (inducible) or nNOS (neuronal) are of great interest for decoding neurodestructive key factors, and (18)F-labelled analogues would allow investigating the NOS-function by molecular imaging with positron emission tomography. Especially, the highly selective nNOS inhibitor 6-((3-((3-fluorophenethylamino)methyl)phenoxy)methyl)-4-methylpyridin-2-amine (10) lends itself as suitable compound to be (18)F-labelled in no-carrier-added (n.c.a.) form. For preparation of the (18)F-labelled nNOS-Inhibitor [(18)F]10 a "build-up" radiosynthesis was developed based on a corresponding iodonium ylide as labelling precursor. The such activated phenethyl group of the compound was efficiently and regioselectively labelled with n.c.a. [(18)F]fluoride in 79% radiochemical yield (RCY). After conversion by reductive amination and microwave assisted displacement of the protecting groups, the desired nNOS-inhibitor was obtained in about 15% total RCY. Alternatively, for a simplified "late-stage" (18)F-labelling procedure a corresponding boronic ester precursor was synthesized and successfully used in a newer, copper(II) mediated n.c.a. (18)F-fluoro-deboroniation reaction, achieving the same total RCY. Thus, both methods proved comparatively suited to provide the highly selective NOS-inhibitor [(18)F]10 as probe for preclinical in vivo studies. PMID:27598109

  4. Synthesis of a Potent Aminopyridine-Based nNOS-Inhibitor by Two Recent No-Carrier-Added (18)F-Labelling Methods.

    PubMed

    Drerup, Christian; Ermert, Johannes; Coenen, Heinz H

    2016-09-01

    Nitric oxide (NO), an important multifunctional signaling molecule, is produced by three isoforms of NO-synthase (NOS) and has been associated with neurodegenerative disorders. Selective inhibitors of the subtypes iNOS (inducible) or nNOS (neuronal) are of great interest for decoding neurodestructive key factors, and (18)F-labelled analogues would allow investigating the NOS-function by molecular imaging with positron emission tomography. Especially, the highly selective nNOS inhibitor 6-((3-((3-fluorophenethylamino)methyl)phenoxy)methyl)-4-methylpyridin-2-amine (10) lends itself as suitable compound to be (18)F-labelled in no-carrier-added (n.c.a.) form. For preparation of the (18)F-labelled nNOS-Inhibitor [(18)F]10 a "build-up" radiosynthesis was developed based on a corresponding iodonium ylide as labelling precursor. The such activated phenethyl group of the compound was efficiently and regioselectively labelled with n.c.a. [(18)F]fluoride in 79% radiochemical yield (RCY). After conversion by reductive amination and microwave assisted displacement of the protecting groups, the desired nNOS-inhibitor was obtained in about 15% total RCY. Alternatively, for a simplified "late-stage" (18)F-labelling procedure a corresponding boronic ester precursor was synthesized and successfully used in a newer, copper(II) mediated n.c.a. (18)F-fluoro-deboroniation reaction, achieving the same total RCY. Thus, both methods proved comparatively suited to provide the highly selective NOS-inhibitor [(18)F]10 as probe for preclinical in vivo studies.

  5. 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. PMID:23412765

  6. 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.

  7. Label-free high-throughput and real-time detections of protein interactions by oblique-incidence reflectivity difference method

    NASA Astrophysics Data System (ADS)

    He, LiPing; Liu, Shuang; Dai, Jun; Lü, HuiBin; Jin, KuiJuan; Yang, GuoZhen

    2014-04-01

    Selected Mouse IgG of 1 mg/mL as target was fabricated on microarray for 500 sample dots. Label-free and real-time reaction dynamic processes were detected between the microarrays with Goat Anti-mouse IgG of 0.02 mg/mL using the oblique-incidence reflectivity difference (OIRD) method. We obtained the reaction results and the reaction dynamic curves of 500 protein dots. In addition, we also used label-free detection of protein microarrays of 10080 sample dots, including BSA and different concentrations of Mouse IgG and Rabbit IgG, by OIRD. The obtained reaction results between the protein microarray with 1 mg/mL Goat Anti-mouse IgG and 1 mg/mL Goat Anti-rabbit IgG are reported herein. Experimental results show that OIRD can be not only label-free high-throughput detection method for biological microarrays but also label-free real-time detection in the interaction processes of biomolecules.

  8. Method for evaluating the potential of C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometry.

    PubMed

    Janle, Elsa M; Lila, Mary Ann; Grannan, Michael; Wood, Lauren; Higgins, Aine; Yousef, Gad G; Rogers, Randy B; Kim, Helen; Jackson, George S; Weaver, Connie M

    2010-04-01

    Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. (14)C-labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions.To study the pharmacokinetics and tissue distribution of (14)C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine (14)C in blood and ISF with a β-counter. However, brain microdialysate samples (14)C levels on the order of 10(7) atoms/sample required AMS technology. The Brain Microdialysate(AUC)/Serum(AUC) ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain. PMID:20419067

  9. Method for evaluating the potential of 14C labeled plant polyphenols to cross the blood-brain barrier using accelerator mass spectrometry

    NASA Astrophysics Data System (ADS)

    Janle, Elsa M.; Lila, Mary Ann; Grannan, Michael; Wood, Lauren; Higgins, Aine; Yousef, Gad G.; Rogers, Randy B.; Kim, Helen; Jackson, George S.; Weaver, Connie M.

    2010-04-01

    Bioactive compounds in botanicals may be beneficial in preventing age-related neurodegenerative diseases, but for many compounds conventional methods may be inadequate to detect if these compounds cross the blood-brain barrier or to track the pharmacokinetics in the brain. By combining a number of unique technologies it has been possible to utilize the power of AMS to study the pharmacokinetics of bioactive compounds in the brain at very low concentrations. 14C labeled compounds can be biosynthesized by plant cell suspension cultures co-incubated with radioisotopically-labeled sucrose and isolated and separated into a series of bioactive fractions. To study the pharmacokinetics and tissue distribution of 14C labeled plant polyphenols, rats were implanted with jugular catheters, subcutaneous ultrafiltration probes and brain microdialysis probes. Labeled fractions were dosed orally. Interstitial fluid (ISF) and brain microdialysate samples were taken in tandem with blood samples. It was often possible to determine 14C in blood and ISF with a β-counter. However, brain microdialysate samples 14C levels on the order of 10 7 atoms/sample required AMS technology. The Brain Microdialysate AUC/Serum AUC ranged from .021- to .029, with the higher values for the glycoside fractions. By using AMS in combination with traditional methods, it is possible to study uptake by blood, distribution to ISF and determine the amount of a dose which can reach the brain and follow the pharmacokinetics in the brain.

  10. MaXIC-Q Web: a fully automated web service using statistical and computational methods for protein quantitation based on stable isotope labeling and LC-MS.

    PubMed

    Tsou, Chih-Chiang; Tsui, Yin-Hao; Yian, Yi-Hwa; Chen, Yi-Ju; Yang, Han-Yin; Yu, Chuan-Yih; Lynn, Ke-Shiuan; Chen, Yu-Ju; Sung, Ting-Yi; Hsu, Wen-Lian

    2009-07-01

    Isotope labeling combined with liquid chromatography-mass spectrometry (LC-MS) provides a robust platform for analyzing differential protein expression in proteomics research. We present a web service, called MaXIC-Q Web (http://ms.iis.sinica.edu.tw/MaXIC-Q_Web/), for quantitation analysis of large-scale datasets generated from proteomics experiments using various stable isotope-labeling techniques, e.g. SILAC, ICAT and user-developed labeling methods. It accepts spectral files in the standard mzXML format and search results from SEQUEST, Mascot and ProteinProphet as input. Furthermore, MaXIC-Q Web uses statistical and computational methods to construct two kinds of elution profiles for each ion, namely, PIMS (projected ion mass spectrum) and XIC (extracted ion chromatogram) from MS data. Toward accurate quantitation, a stringent validation procedure is performed on PIMSs to filter out peptide ions interfered with co-eluting peptides or noise. The areas of XICs determine ion abundances, which are used to calculate peptide and protein ratios. Since MaXIC-Q Web adopts stringent validation on spectral data, it achieves high accuracy so that manual validation effort can be substantially reduced. Furthermore, it provides various visualization diagrams and comprehensive quantitation reports so that users can conveniently inspect quantitation results. In summary, MaXIC-Q Web is a user-friendly, interactive, robust, generic web service for quantitation based on ICAT and SILAC labeling techniques.

  11. Wearable cameras can reduce dietary under-reporting: doubly labelled water validation of a camera-assisted 24 h recall.

    PubMed

    Gemming, Luke; Rush, Elaine; Maddison, Ralph; Doherty, Aiden; Gant, Nicholas; Utter, Jennifer; Ni Mhurchu, Cliona

    2015-01-28

    Preliminary research has suggested that wearable cameras may reduce under-reporting of energy intake (EI) in self-reported dietary assessment. The aim of the present study was to test the validity of a wearable camera-assisted 24 h dietary recall against the doubly labelled water (DLW) technique. Total energy expenditure (TEE) was assessed over 15 d using the DLW protocol among forty adults (n 20 males, age 35 (sd 17) years, BMI 27 (sd 4) kg/m2 and n 20 females, age 28 (sd 7) years, BMI 22 (sd 2) kg/m2). EI was assessed using three multiple-pass 24 h dietary recalls (MP24) on days 2-4, 8-10 and 13-15. On the days before each nutrition assessment, participants wore an automated wearable camera (SenseCam (SC)) in free-living conditions. The wearable camera images were viewed by the participants following the completion of the dietary recall, and their changes in self-reported intakes were recorded (MP24+SC). TEE and EI assessed by the MP24 and MP24+SC methods were compared. Among men, the MP24 and MP24+SC measures underestimated TEE by 17 and 9%, respectively (P< 0.001 and P= 0.02). Among women, these measures underestimated TEE by 13 and 7%, respectively (P< 0.001 and P= 0.004). The assistance of the wearable camera (MP24+SC) reduced the magnitude of under-reporting by 8% for men and 6% for women compared with the MP24 alone (P< 0.001 and P< 0.001). The increase in EI was predominantly from the addition of 265 unreported foods (often snacks) as revealed by the participants during the image review. Wearable cameras enhance the accuracy of self-report by providing passive and objective information regarding dietary intake. High-definition image sensors and increased imaging frequency may improve the accuracy further.

  12. Metabolic power of European starlings Sturnus vulgaris during flight in a wind tunnel, estimated from heat transfer modelling, doubly labelled water and mask respirometry.

    PubMed

    Ward, S; Möller, U; Rayner, J M V; Jackson, D M; Nachtigall, W; Speakman, J R

    2004-11-01

    It is technically demanding to measure the energetic cost of animal flight. Each of the previously available techniques has some disadvantage as well advantages. We compared measurements of the energetic cost of flight in a wind tunnel by four European starlings Sturnus vulgaris made using three independent techniques: heat transfer modelling, doubly labelled water (DLW) and mask respirometry. We based our heat transfer model on thermal images of the surface temperature of the birds and air flow past the body and wings calculated from wing beat kinematics. Metabolic power was not sensitive to uncertainty in the value of efficiency when estimated from heat transfer modelling. A change in the assumed value of whole animal efficiency from 0.19 to 0.07 (the range of estimates in previous studies) only altered metabolic power predicted from heat transfer modelling by 13%. The same change in the assumed value of efficiency would cause a 2.7-fold change in metabolic power if it were predicted from mechanical power. Metabolic power did not differ significantly between measurements made using the three techniques when we assumed an efficiency in the range 0.11-0.19, although the DLW results appeared to form a U-shaped power-speed curve while the heat transfer model and respirometry results increased linearly with speed. This is the first time that techniques for determining metabolic power have been compared using data from the same birds flying under the same conditions. Our data provide reassurance that all the techniques produce similar results and suggest that heat transfer modelling may be a useful method for estimating metabolic rate.

  13. Metabolic power of European starlings Sturnus vulgaris during flight in a wind tunnel, estimated from heat transfer modelling, doubly labelled water and mask respirometry.

    PubMed

    Ward, S; Möller, U; Rayner, J M V; Jackson, D M; Nachtigall, W; Speakman, J R

    2004-11-01

    It is technically demanding to measure the energetic cost of animal flight. Each of the previously available techniques has some disadvantage as well advantages. We compared measurements of the energetic cost of flight in a wind tunnel by four European starlings Sturnus vulgaris made using three independent techniques: heat transfer modelling, doubly labelled water (DLW) and mask respirometry. We based our heat transfer model on thermal images of the surface temperature of the birds and air flow past the body and wings calculated from wing beat kinematics. Metabolic power was not sensitive to uncertainty in the value of efficiency when estimated from heat transfer modelling. A change in the assumed value of whole animal efficiency from 0.19 to 0.07 (the range of estimates in previous studies) only altered metabolic power predicted from heat transfer modelling by 13%. The same change in the assumed value of efficiency would cause a 2.7-fold change in metabolic power if it were predicted from mechanical power. Metabolic power did not differ significantly between measurements made using the three techniques when we assumed an efficiency in the range 0.11-0.19, although the DLW results appeared to form a U-shaped power-speed curve while the heat transfer model and respirometry results increased linearly with speed. This is the first time that techniques for determining metabolic power have been compared using data from the same birds flying under the same conditions. Our data provide reassurance that all the techniques produce similar results and suggest that heat transfer modelling may be a useful method for estimating metabolic rate. PMID:15531650

  14. 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)

  15. 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.

  16. 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.

  17. Extraction of water labeled with oxygen 15 during single-capillary transit. Influence of blood pressure, osmolarity, and blood-brain barrier damage

    SciTech Connect

    Go, K.G.; Lammertsma, A.A.; Paans, A.M.; Vaalburg, W.; Woldring, M.G.

    1981-09-01

    By external detection, the influence of arterial blood pressure (BP), osmolarity, and cold-induced blood-brain barrier damage was assessed on the extraction of water labeled with oxygen 15 during single-capillary transit in the rat. There was an inverse relation between arterial BP and extraction that was attributable to the influence of arterial BP on cerebral blood flow (CBF) and the relation between CBF and extraction. Neither arterial BP nor osmolarity of the injected bolus had any direct effect on extraction of water 15O, signifying that the diffusional exchange component (determined by blood flow) of extraction greatly surpasses the convection flow contribution by hydrostatic or osmotic forces. Damage to the blood-brain barrier did not change its permeability to water.

  18. General method for labeling siRNA by click chemistry with fluorine-18 for the purpose of PET imaging.

    PubMed

    Mercier, Frédéric; Paris, Jérôme; Kaisin, Geoffroy; Thonon, David; Flagothier, Jessica; Teller, Nathalie; Lemaire, Christian; Luxen, André

    2011-01-19

    The alkyne-azide Cu(I)-catalyzed Huisgen cycloaddition, a click-type reaction, was used to label a double-stranded oligonucleotide (siRNA) with fluorine-18. An alkyne solid support CPG for the preparation of monostranded oligonucleotides functionalized with alkyne has been developed. Two complementary azide labeling agents (1-(azidomethyl)-4-[(18)F]fluorobenzene) and 1-azido-4-(3-[(18)F]fluoropropoxy)benzene have been produced with 41% and 35% radiochemical yields (decay-corrected), respectively. After annealing with the complementary strand, the siRNA was directly labeled by click chemistry with [(18)F]fluoroazide to produce the [(18)F]-radiolabeled siRNA with excellent radiochemical yield and purity.

  19. Method of sealing an ultracapacitor substantially free of water

    DOEpatents

    Chapman-Irwin, Patricia; Feist, Thomas Paul

    2002-04-02

    A method of sealing an ultracapacitor substantially free of water is disclosed. The method includes providing a multilayer cell comprising two solid, non porous current collectors, separated by two porous electrodes with a separator between the two electrodes, sealing the cell with a reclosable hermetic closure. Water inside the closure is dissociated by an applied voltage to the cell and escapes in the form of hydrogen and oxygen when the closure is unmated, the closure is then mated to hermetically seal the cell which is substantially free of water.

  20. Method to detect the end-point for PCR DNA amplification using an ionically labeled probe and measuring impedance change

    DOEpatents

    Miles, Robin R.; Belgrader, Phillip; Fuller, Christopher D.

    2007-01-02

    Impedance measurements are used to detect the end-point for PCR DNA amplification. A pair of spaced electrodes are located on a surface of a microfluidic channel and an AC or DC voltage is applied across the electrodes to produce an electric field. An ionically labeled probe will attach to a complementary DNA segment, and a polymerase enzyme will release the ionic label. This causes the conductivity of the solution in the area of the electrode to change. This change in conductivity is measured as a change in the impedance been the two electrodes.

  1. Biological Stability of Drinking Water: Controlling Factors, Methods, and Challenges.

    PubMed

    Prest, Emmanuelle I; Hammes, Frederik; van Loosdrecht, Mark C M; Vrouwenvelder, Johannes S

    2016-01-01

    Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order

  2. Biological Stability of Drinking Water: Controlling Factors, Methods, and Challenges

    PubMed Central

    Prest, Emmanuelle I.; Hammes, Frederik; van Loosdrecht, Mark C. M.; Vrouwenvelder, Johannes S.

    2016-01-01

    Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order

  3. A discovery of an ultra-pure water detection method based on water mark

    NASA Astrophysics Data System (ADS)

    Cao, Hui-Wen; Jing, Yu-Peng; Zhao, Shi-Rui; Xu, Xin-Wei; Tian, He; Xin, Xin; Li, Xiao-Ning; Liu, Bo; Liu, Rui-Tao; Wang, Gang; Ge, Jie; Cai, Hua-Lin; Yang, Yi; Ren, Tian-Ling

    2015-02-01

    The purity evaluation of deionized (DI) water is highly desirable for VLSI or ULSI industry, as the traditional "reverse osmosis filter" cannot always meet the requirement towards the DI water. The filtered DI water may still contain many contaminations which are not up to the standard for the wet cleaning of wafer surface. A novel method is presented by analyzing the residues of a water droplet after the low-temperature evaporation. The contamination contained in the water will remain during the gasification. By analyzing the residual contamination's morphology, the purity of the DI water can be estimated by employing merely a 3D laser microscope. Compared to the traditional fluorescence detecting system for water quality monitoring, it is simpler and has a lower cost. The paper describes an excellent water detection method which is meaningful for preparing ultra-pure water. Experimental results have shown that the deionized distilled (DID) water can repeatedly get a higher purity using this detection method. The DID water can be applied to the wet cleaning of wafer surface, preparation of chemical reagents and many other aspects.

  4. A simple method for locating the fresh water-salt water interface using pressure data.

    PubMed

    Kim, Kue-Young; Chon, Chul-Min; Park, Ki-Hwa

    2007-01-01

    Salt water intrusion is a key issue in dealing with exploitation, restoration, and management of fresh ground water in coastal aquifers. Constant monitoring of the fresh water-salt water interface is necessary for proper management of ground water resources. This study presents a simple method to estimate the depth of the fresh water-salt water interface in coastal aquifers using two sets of pressure data obtained from the fresh and saline zones within a single borehole. This method uses the density difference between fresh water and saline water and can practically be used at coastal aquifers that have a relatively sharp fresh water-salt water interface with a thin transition zone. The proposed method was applied to data collected from a coastal aquifer on Jeju Island, Korea, to estimate the variations in the depth of the interface. The interface varied with daily tidal fluctuations and heavy rainfall in the rainy season. The estimated depth of the interface showed a good agreement with the measured electrical conductivity profile.

  5. Ultrastructural localization of wheat germ agglutinin binding sites on the sperm surface of water buffalo (Bubalus bubalis). A fracture label study.

    PubMed

    Bains, H K; Pabst, M A; Werner, G; Bawa, S R

    1993-10-01

    In the present study we have examined the plasma membrane surface organization employing fluorescein isothiocyanate linked wheat germ agglutinin (WGA) of the cauda epididymal and ejaculated spermatozoa of water buffalo. Intramembrane particle distribution pattern in the various segments of the spermatozoa has also been observed. WGA-ovomucoid gold has been used to study the distribution of sialoproteins on the sperm surface. With fracture label, WGA receptor sites have been identified on the fractured membrane halves of the sperm plasma membrane overlying the acrosome as well as the middle piece and the principle piece.

  6. Dosimetry of intravenously administered oxygen-15 labelled water in man: a model based on experimental human data from 21 subjects.

    PubMed

    Smith, T; Tong, C; Lammertsma, A A; Butler, K R; Schnorr, L; Watson, J D; Ramsay, S; Clark, J C; Jones, T

    1994-10-01

    Models based on uniform distribution of tracer in total body water underestimate the absorbed dose from H2(15)O because of the short half-life (2.04 min) of 15O, which leads to non-uniform distribution of absorbed dose and also complicates the direct measurement of organ retention curves. However, organ absorbed doses can be predicted by the present kinetic model based on the convolution technique. The measured time course of arterial H2(15)O concentration following intravenous administration represents the input function to organs. The impulse response of a given organ is its transit time function determined by blood flow and the partition of water between tissue and blood. Values of these two parameters were taken from the literature. Integrals of the arterial input function and organ transit time functions were used to derive integrals of organ retention functions (organ residence times). The latter were used with absorbed dose calculation software (MIRDOSE-2) to obtain estimates for 24 organs. From the mean values of organ absorbed doses, the effective dose equivalent (EDE) and effective dose (ED) were calculated. From measurements on 21 subjects, the average value for both EDE and ED was calculated to be 1.2 microSv.MBq-1 compared with a value of about 0.5 microSv.MBq-1 predicted by uniform water distribution models. Based on the human data, a method of approximating H2(15)O absorbed dose values from body surface area is described. PMID:7828623

  7. Methods for the determination of organic compounds in drinking water

    SciTech Connect

    Not Available

    1988-12-01

    Thirteen analytical methods for the identification and measurement of organic compounds in drinking water are described in detail. Six of the methods are for volatile organic compounds (VOCs) and certain disinfection by-products. These methods were cited in the Federal Register of July 8, 1987, under the National Primary Drinking Water Regulations. The other seven methods are designed for the determination of a variety of synthetic organic compounds and pesticides, and these methods were cited in proposed drinking-water regulations in the Federal Register of May 22, 1989. Five of the methods utilize the inert gas purge-and-trap extraction procedure for VOCs, six methods employ a classical liquid-liquid extraction, one method uses a new liquid-solid extraction technique, and one method is for direct aqueous analysis. Of the 13 methods, 12 use either packed or capillary gas-chromatography column separations followed by detection with mass spectrometry or a selective gas-chromatography detector. One method is based on a high-performance liquid-chromatography separation.

  8. A CCG-Based Method for Training a Semantic Role Labeler in the Absence of Explicit Syntactic Training Data

    ERIC Educational Resources Information Center

    Boxwell, Stephen A.

    2011-01-01

    Treebanks are a necessary prerequisite for many NLP tasks, including, but not limited to, semantic role labeling. For many languages, however, treebanks are either nonexistent or too small to be useful. Time-critical applications may require rapid deployment of natural language software for a new critical language--much faster than the development…

  9. Tectonic influences on ground water quality: insight from complementary methods.

    PubMed

    Earman, Sam; McPherson, Brian J O L; Phillips, Fred M; Ralser, Steve; Herrin, James M; Broska, James

    2008-01-01

    A study using multiple techniques provided insight into tectonic influences on ground water systems; the results can help to understand ground water systems in the tectonically active western United States and other parts of the world. Ground water in the San Bernardino Valley (Arizona, United States and Sonora, Mexico) is the main source of water for domestic use, cattle ranching (the primary industry), and the preservation of threatened and endangered species. To improve the understanding of ground water occurrence, movement, and sustainability, an investigation was conducted using a number of complementary methods, including major ion geochemistry, isotope hydrology, analysis of gases dissolved in ground water, aquifer testing, geophysics, and an examination of surface and subsurface geology. By combining information from multiple lines of investigation, a more complete picture of the basin hydrogeology was assembled than would have been possible using fewer methods. The results show that the hydrogeology of the San Bernardino Valley is markedly different than that of its four neighboring basins in the United States. The differences include water quality, chemical evolution, storage, and residence time. The differences result from the locally unique geology of the San Bernardino Valley, which is due to the presence of a magmatically active accommodation zone (a zone separating two regions of normal faults with opposite dips). The geological differences and the resultant hydrological differences between the San Bernardino Valley and its neighboring basins may serve as a model for the distinctive nature of chemical evolution of ground water in other basins with locally distinct tectonic histories.

  10. The Halliwick Method: Water Freedom for Individuals with Disabilities.

    ERIC Educational Resources Information Center

    Grosse, Susan J.

    This publication explains the Halliwick Method of swim instruction for people with disabilities. The Halliwick Method emphasizes independent functioning in the water, obtained through the development of control over body movements and establishing physical and psychological comfort. Containing over 75 photographs, including underwater shots, this…

  11. GROUND WATER PURGING AND SAMPLING METHODS: HISTORY VS. HYSTERIA

    EPA Science Inventory

    It has been over 10 years since the low-flow ground water purging and sampling method was initially reported in the literature. The method grew from the recognition that well purging was necessary to collect representative samples, bailers could not achieve well purging, and high...

  12. Laser labeling, a safe technology to label produce

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laser labeling of fruits and vegetables is an alternative means to label produce. Low energy CO2 laser beams etch the surface showing the contrasting underlying layer. These etched surfaces can promote water loss and potentially allow for entry of decay organisms. The long-term effects of laser labe...

  13. Learning with imperfectly labeled patterns

    NASA Technical Reports Server (NTRS)

    Chittineni, C. B.

    1979-01-01

    The problem of learning in pattern recognition using imperfectly labeled patterns is considered. The performance of the Bayes and nearest neighbor classifiers with imperfect labels is discussed using a probabilistic model for the mislabeling of the training patterns. Schemes for training the classifier using both parametric and non parametric techniques are presented. Methods for the correction of imperfect labels were developed. To gain an understanding of the learning process, expressions are derived for success probability as a function of training time for a one dimensional increment error correction classifier with imperfect labels. Feature selection with imperfectly labeled patterns is described.

  14. Label-free, real-time detection of the dynamic processes of protein degradation using oblique-incidence reflectivity difference method

    NASA Astrophysics Data System (ADS)

    Liu, S.; Zhu, J. H.; He, L. P.; Dai, J.; Lu, H. B.; Wu, L.; Jin, K. J.; Yang, G. Z.; Zhu, H.

    2014-04-01

    Based on the requirements for studying the dynamic process of proteinase action substrates in life science, we selected six random proteins including 1L-10, SCGB2A2, CENPQ, GST, HK1, KLHL7, as well as five different concentrations of 1L-10 proteins of 1 mg/ml, 0.5 mg/ml, 0.25 mg/ml, 0.125 mg/ml, and 0.0625 mg/ml, and fabricated two types of substrate protein microarrays, respectively. We detected the dynamic processes of proteins degraded by proteinase K using oblique-incidence reflectivity difference (OIRD) method in a label-free and real-time manner. We obtained the relevant degradation velocities and the degradation time. The experimental results demonstrate that OIRD has the ability to study proteinase action substrates which is out of reach of label methods and is expected to offer opportunities to determine protease-substrate relationships on the systems biology level.

  15. A method to extract soil water for stable isotope analysis

    USGS Publications Warehouse

    Revesz, K.; Woods, P.H.

    1990-01-01

    A method has been developed to extract soil water for determination of deuterium (D) and 18O content. The principle of this method is based on the observation that water and toluene form an azeotropic mixture at 84.1??C, but are completely immiscible at ambient temperature. In a specially designed distillation apparatus, the soil water is distilled at 84.1??C with toluene and is separated quantitatively in the collecting funnel at ambient temperature. Traces of toluene are removed and the sample can be analyzed by mass spectrometry. Kerosene may be substituted for toluene. The accuracy of this technique is ?? 2 and ?? 0.2???, respectively, for ??D and ??18O. Reduced accuracy is obtained at low water contents. ?? 1990.

  16. Apparatus and a method for biological treatment of waste waters

    SciTech Connect

    Besik, F.

    1983-12-20

    An apparatus and a method for biological treatment of waste waters achieving biological oxidation of organic matter, biological nitrification and denitrification of nitrogenous compounds and biological removal of phosphorus and clarification of the treated waste water in a single reaction tank in a single suspended growth sludge system without the use of traditional compressors, mixers, recirculation pumps, piping and valving and without the use of the traditional clarifier.

  17. Characterization of biodegradation intermediates of nonionic surfactants by MALDI-MS. 2. Oxidative biodegradation profiles of uniform octylphenol polyethoxylate in 18O-labeled water.

    PubMed

    Sato, Hiroaki; Shibata, Atsushi; Wang, Yang; Yoshikawa, Hiromichi; Tamura, Hiroto

    2003-01-01

    This paper reports the characterization of the biodegradation intermediates of octylphenol octaethoxylate (OP(8)EO) by means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The biodegradation test study was carried out in a pure culture (Pseudomonas putida S-5) under aerobic conditions using OP(8)EO as the sole carbon source and (18)O-labeled water as an incubation medium. In the MALDI-MS spectra of biodegraded samples, a series of OP(n)EO molecules with n = 2-8 EO units and their corresponding carboxylic acid products (OP(n)EC) were observed. The use of purified OP(8)EO enabled one to distinguish the shortened OPEO molecules as biodegradation intermediates. Furthermore, the formation of OP(8)EC (the oxidized product of OP(8)EO) supported the notion that terminal oxidation is a step in the biodegradation process. When biodegradation study was carried out in (18)O-labeled water, incorporation of (18)O atoms into the carboxyl group was observed for OPEC, while no incorporation was observed for the shortened OPEO products. These results could provide some rationale to the biodegradation mechanism of alkylphenol polyethoxylates. PMID:12523845

  18. Investigations into water mitigation using a meshless particle method

    NASA Astrophysics Data System (ADS)

    Liu, M. B.; Liu, G. R.; Lam, K. Y.

    It is very difficult for traditional numerical methods to simulate the problems of water mitigation which has been increasingly used to reduce blast effects. This paper studies water mitigation problems by using smoothed particle hydrodynamics (SPH), which is a meshless, Lagrangian method appealing in treating large deformation explosion events with significant inhomogeneities. Numerical verifications considering high explosive detonation and underwater explosion shock waves have demonstrated the effectiveness of the SPH method, the solution procedure and the code. Contact and non-contact water mitigation simulations have been carried out and are compared with the case without mitigation. For either contact or non-contact water shield, the peak shock pressure and the equilibrium gas pressure are reduced to different levels according to the relevant geometry of the system setup. An optimum contact water shield thickness is found to produce the best mitigation effect for a given high explosive charge, while the non-contact water shield, if properly designed, can result in further reduction of the peak shock pressure and the equilibrium gas pressure.

  19. Quantitation of a spin polarization-induced nuclear Overhauser effect (SPINOE) between a hyperpolarized 13C-labeled cell metabolite and water protons

    PubMed Central

    Marco-Rius, Irene; Bohndiek, Sarah E; Kettunen, Mikko I; Larkin, Timothy J; Basharat, Meer; Seeley, Colm; Brindle, Kevin M

    2014-01-01

    The spin polarization-induced nuclear Overhauser effect (SPINOE) describes the enhancement of spin polarization of solvent nuclei by the hyperpolarized spins of a solute. In this communication we demonstrate that SPINOEs can be observed between [1,4-13C2]fumarate, hyperpolarized using the dissolution dynamic nuclear polarization technique, and solvent water protons. We derive a theoretical expression for the expected enhancement and demonstrate that this fits well with experimental measurements. Although the magnitude of the effect is relatively small (around 2% measured here), the SPINOE increases at lower field strengths, so that at clinically relevant magnetic fields (1.5–3 T) it may be possible to track the passage through the circulation of a bolus containing a hyperpolarized 13C-labeled substrate through the increase in solvent water 1H signal. © 2014 The Authors. Contrast Media & Molecular Imaging published by John Wiley and Sons, Ltd. PMID:24523064

  20. Water footprint assessment for wastewater treatment: method, indicator, and application.

    PubMed

    Shao, Ling; Chen, G Q

    2013-07-16

    The water footprint in terms of the sum of both direct and indirect water cost of wastewater treatment is for the first time accounted in this work. On the basis of the hybrid method as a combination of process analysis and input-output analysis, a detailed water footprint accounting procedure is provided to cover the supply chain of a wastewater treatment plant. A set of indices intending to reveal the efficiency as well as renewability of wastewater treatment systems are devised as parallels of corresponding indicators in net energy analysis for energy supply systems. A case study is carried out for the Beijing Space City wastewater treatment plant as a landmark project. The high WROI (water return on investment) and low WIWP (water investment in water purified) indicate a high efficiency and renewability of the case system, illustrating the fundamental function of wastewater treatment for water reuse. The increasing of the wastewater and sludge treatment rates are revealed in an urgent need to reduce the water footprint of China and to improve the performance of wastewater treatment.

  1. Isolation of Legionella from water samples using various culture methods.

    PubMed

    Kusnetsov, J M; Jousimies-Somer, H R; Nevalainen, A I; Martikainen, P J

    1994-02-01

    The efficacy of a non-selective medium and two selective media were compared for the isolation of legionellas from water samples. The effect of acid wash treatment for decontamination of the water samples on the isolation frequency of legionellas was also studied. The 236 samples were taken from cooling, humidifying and drinking water systems; 21% were legionella-positive when inoculated directly on modified Wadowsky-Yee (MWY) medium and 26% were positive when concentrated (x 200) before cultivation on MWY or CCVC media. Inoculation on MWY medium after concentration followed by decontamination by the acid-wash technique gave the highest isolation frequency (31%). The lowest frequency (8%) was found with the non-selective BCYE alpha medium. An isolation frequency of 28% was achieved with the BCYE alpha medium after concentration and acid-wash treatment of the samples. Forty per cent of the samples were positive for legionellas when the results from all the culture methods were combined. Not all the legionella-positive samples were identified by a single culture method. Ninety-three of the 95 positive samples were detected with the two best combinations of three culture methods. The best culture method for detecting legionellas depended on the source of the water sample. Some water quality characteristics, like temperature and organic matter content, affected the isolation frequency of Legionella spp.

  2. General Characterization Methods for Photoelectrochemical Cells for Solar Water Splitting.

    PubMed

    Shi, Xinjian; Cai, Lili; Ma, Ming; Zheng, Xiaolin; Park, Jong Hyeok

    2015-10-12

    Photoelectrochemical (PEC) water splitting is a very promising technology that converts water into clean hydrogen fuel and oxygen by using solar light. However, the characterization methods for PEC cells are diverse and a systematic introduction to characterization methods for PEC cells has rarely been attempted. Unlike most other review articles that focus mainly on the material used for the working electrodes of PEC cells, this review introduces general characterization methods for PEC cells, including their basic configurations and methods for characterizing their performance under various conditions, regardless of the materials used. Detailed experimental operation procedures with theoretical information are provided for each characterization method. The PEC research area is rapidly expanding and more researchers are beginning to devote themselves to related work. Therefore, the content of this Minireview can provide entry-level knowledge to beginners in the area of PEC, which might accelerate progress in this area.

  3. General Characterization Methods for Photoelectrochemical Cells for Solar Water Splitting.

    PubMed

    Shi, Xinjian; Cai, Lili; Ma, Ming; Zheng, Xiaolin; Park, Jong Hyeok

    2015-10-12

    Photoelectrochemical (PEC) water splitting is a very promising technology that converts water into clean hydrogen fuel and oxygen by using solar light. However, the characterization methods for PEC cells are diverse and a systematic introduction to characterization methods for PEC cells has rarely been attempted. Unlike most other review articles that focus mainly on the material used for the working electrodes of PEC cells, this review introduces general characterization methods for PEC cells, including their basic configurations and methods for characterizing their performance under various conditions, regardless of the materials used. Detailed experimental operation procedures with theoretical information are provided for each characterization method. The PEC research area is rapidly expanding and more researchers are beginning to devote themselves to related work. Therefore, the content of this Minireview can provide entry-level knowledge to beginners in the area of PEC, which might accelerate progress in this area. PMID:26365789

  4. Autoradiographic detection of (/sup 125/I)-secondary antiserum: a sensitive light and electron microscopic labeling method compatible with peroxidase immunocytochemistry for dual localization of neuronal antigens

    SciTech Connect

    Pickel, V.M.; Chan, J.; Milner, T.A.

    1986-06-01

    We examined whether autoradiographic localization of (/sup 125/I)-antirabbit immunoglobulin (IgG) was suitable for light and electron microscopic detection of a rabbit antiserum to the catecholamine-synthesizing enzyme, tyrosine hydroxylase (TH), and whether autoradiographic and peroxidase labeling could be combined for simultaneous immunocytochemical identification of TH and neuropeptides in brain. Adult rat brains were fixed by aortic arch perfusion with acrolein and paraformaldehyde. Vibratome sections of the fixed tissues were incubated with various dilutions of TH antiserum followed by (/sup 125/I)-secondary IgG. These sections were then directly processed for autoradiography or were incubated with rabbit antiserum to substance P (SP) or methionine (Met5)-enkephalin (ME). These latter sections were then processed by the peroxidase-antiperoxidase (PAP) or conjugated peroxidase methods followed by autoradiography. Exposure periods of 12-20 days for light microscopy or 90 days for electron microscopy yielded substantial accumulations of silver grains even at the highest (1:30,000) dilution of TH antiserum. At this dilution, immunoreactivity for TH was virtually nondetectable by PAP and conjugated peroxidase methods. The differential sensitivities of the autoradiographic versus peroxidase methods provided a means for separable identification of rabbit antiserum to TH and to SP or ME. Ultrastructural analysis of the catecholaminergic neurons in the medial nuclei of the solitary tract (NTS) showed selective cytoplasmic localization of silver grains for (/sup 125/I)-labeling of TH in perikarya, dendrites, and terminals. Within single thin sections prepared for dual labeling, the peroxidase marker for SP and for ME was differentially localized with respect to autoradiographic labeling of TH.

  5. Appliance energy labeling takes effect

    SciTech Connect

    Not Available

    1980-06-01

    Consumers buying household appliances will be helped by energy-efficiency labels and minimum efficiency standards required for refrigerators and refrigerator/freezers, freezers, dishwashers, water heaters, clothes washers, room air conditioners, and furnaces. The ENERGYGUIDE labels must be displayed in the store and in catalogs. Two voluntary efficiency programs were combined in the Energy Policy and Conservation Act (EPCA) requiring labels by 1980. Shoppers may compare the efficiencies of appliances and compute the actual cost differential over the lifetime of the equipment. Manufacturers have responded with more-efficient models, but the impact of efficient appliances on energy consumption will be small. A sample label with the required information is illustrated. (DCK)

  6. A method for determining the periodicity of a troponin component in isolated insect flight muscle thin filaments by gold/Fab labelling.

    PubMed

    Newman, R; Butcher, G W; Bullard, B; Leonard, K R

    1992-03-01

    Insect flight muscle has a large component (Tn-H) in the tropomyosin-troponin complex that is not present in vertebrate striated muscle thin filaments. Tn-H is shown by gold/Fab labelling to be present at regular intervals in insect flight muscle thin filaments. The Fab fragment of a monoclonal antibody to Tn-H was conjugated directly with colloidal gold and this probe used to label isolated thin filaments from the flight muscle of Lethocerus indicus (water bug). The distribution of gold particles seen in electron microscope images of negatively stained thin filaments was analysed to show that the probe bound to sites having a periodicity of approximately 40 nm, which is the expected value for the tropomyosin-troponin repeat. Conjugates of Fab with colloidal gold particles of 3 nm diameter labelled almost all sites. Conjugates with gold particles of 5 nm and 10 nm diameter labelled less efficiently (70% and 30%, respectively) but analysis of the distribution of inter-particle intervals among a number of filaments again gave the same fundamental spacing of 40 nm. The error in the measurements (standard deviation approximately +/- 4.2 for 5 nm gold/Fab) is less than earlier estimates for the size of the gold/Fab complex. Measurements on gold/Fab in negative stain suggest that the bound Fab contributes a shell about 2 nm in thickness around the gold particle. The radius of the probe (about 4.5 nm for 5 nm gold/Fab) would then be consistent with the value of error found. The size of the probe suggests that the gold particle binds to the side of the Fab molecule, rather close to the antibody combining site. The potential resolution of the technique may thus be better than originally expected.

  7. Methods for enumerating Escherichia coli in subtropical waters.

    PubMed

    Cheung, W H; Ha, D K; Yeung, K Y; Hung, R P

    1991-04-01

    The standard membrane filtration method of the UK has been modified in order to improve its specificity for enumerating Escherichia coli in the subtropical waters of Hong Kong. This involves incorporating into the membrane lauryl sulphate (mLS) method either an in situ urease test (the mLS-UA method), or an in situ beta-glucuronidase test (the mLS-GUD method). The false-positive errors of the mLS-UA and mLS-GUD methods are low, ranging from 3-5%. A comparison between the membrane filtration (mLS-UA) method and the multiple tube technique in testing E. coli in subtropical beach-waters has demonstrated that the former can give much more precise counts, and is the method of choice for such a purpose. The mLS-GUD method, for which automated counting of E. coli colonies is possible, is a good alternative to mLS-UA in routine enumeration of this bacterial indicator in environmental waters.

  8. [ A method for detection and quantitative analysis of two-dimensional patterns of labeled neuron distribution in the cerebral cortex].

    PubMed

    Merkul'eva, N S; Nikitina, N I

    2010-01-01

    The aim of the present investigation was to elaborate an algorithm for labeled neuron 2-D pattern reconstruction and its analysis. Using the study of cat cortico-cortical connections between the areas 17, 18 and posteromedial lateral suprasylvian area as an example, it was shown that proposed algorithm is adequate for present aims of initial neuron pattern reconstruction and analysis, and that it can be applied as a working algorithm for highly specialized computer program elaboration.

  9. Detection of Legionella pneumophila by PCR-ELISA method in industrial cooling tower water.

    PubMed

    Soheili, Majid; Nejadmoghaddam, Mohammad Reza; Babashamsi, Mohammad; Ghasemi, Jamileh; Jeddi Tehrani, Mahmood

    2007-11-15

    Water supply and Cooling Tower Water (CTW) are among the most common sources of Legionella pneumophila (LP) contamination. A nonradio active method is described to detect LP in industrial CTW samples. DNA was purified and amplified by nested -PCR with amplimers specific for the 16s rRNA gene of LP. The 5' end biotinylated oligomer probe was immobilized on sterptavidin B coated microtiter plates. The nested-PCR product was labeled with digoxigenin and then hybridized with 5'-biotinylated probes. The amplification products were detected by using proxidase-labled anti dioxygenin antibody in a colorimetric reaction. The assay detected LP present in 1 L of 5 CTW samples examined. All of the samples were Legionella positive in both culture and PCR-ELISA methods. The PCR-ELISA assay appears to exhibit high specificity and is a more rapid technique in comparison with bacterial culture method. Thus could prove suitable for use in the routine examination of industrial CTW contamination. PMID:19090273

  10. Comparison of beach bacterial water quality indicator measurement methods.

    PubMed

    Noble, Rachel T; Weisberg, Stephen B; Leecaster, Molly K; McGee, Charles D; Ritter, Kerry; Walker, Kathy O; Vainik, Patricia M

    2003-01-01

    Three methods (membrane filtration, multiple tube fermentation, and chromogenic substrate technology kits manufactured by IDEXX Laboratories, Inc.) are routinely used to measure indicator bacteria for beach water quality. To assess comparability of these methods, quantify within-laboratory variability for each method, and place that variability into context of variability among laboratories using the same method, 22 southern California laboratories participated in a series of intercalibration exercises. Each laboratory processed three to five replicates from thirteen samples, with total coliforms, fecal coliforms or enterococci measured depending on the sample. Results were generally comparable among methods, though membrane filtration appeared to underestimate the other two methods for fecal coliforms, possibly due to clumping. Variability was greatest for the multiple tube fermentation method. For all three methods, within laboratory variability was greater than among laboratories variability.

  11. A novel method for measuring polymer-water partition coefficients.

    PubMed

    Zhu, Tengyi; Jafvert, Chad T; Fu, Dafang; Hu, Yue

    2015-11-01

    Low density polyethylene (LDPE) often is used as the sorbent material in passive sampling devices to estimate the average temporal chemical concentration in water bodies or sediment pore water. To calculate water phase chemical concentrations from LDPE concentrations accurately, it is necessary to know the LDPE-water partition coefficients (KPE-w) of the chemicals of interest. However, even moderately hydrophobic chemicals have large KPE-w values, making direct measurement experimentally difficult. In this study we evaluated a simple three phase system from which KPE-w can be determined easily and accurately. In the method, chemical equilibrium distribution between LDPE and a surfactant micelle pseudo-phase is measured, with the ratio of these concentrations equal to the LDPE-micelle partition coefficient (KPE-mic). By employing sufficient mass of polymer and surfactant (Brij 30), the mass of chemical in the water phase remains negligible, albeit in equilibrium. In parallel, the micelle-water partition coefficient (Kmic-w) is determined experimentally. KPE-w is the product of KPE-mic and Kmic-w. The method was applied to measure values of KPE-w for 17 polycyclic aromatic hydrocarbons, 37 polychlorinated biphenyls, and 9 polybrominated diphenylethers. These values were compared to literature values. Mass fraction-based chemical activity coefficients (γ) were determined in each phase and showed that for each chemical, the micelles and LDPE had nearly identical affinity.

  12. A simplified molecular method for distinguishing among species and ploidy levels in European water frogs (Pelophylax).

    PubMed

    Hauswaldt, J Susanne; Höer, Manuela; Ogielska, Maria; Christiansen, Ditte G; Dziewulska-Szwajkowska, Daria; Czernicka, Elżbieta; Vences, Miguel

    2012-09-01

    Western Palearctic water frogs in the genus Pelophylax are a set of morphologically similar anuran species that form hybridogenetic complexes. Fully reliable identification of species and especially of hybrid ploidy depends on karyological and molecular methods. In central Europe, native water frog populations consist of the Pelophylax esculentus complex, that is, P. lessonae (LL), P. ridibundus (RR) and the hybrid form P. esculentus that can have different karyotypes (RL, LLR and RRL). We developed existing molecular methods further and propose a simple PCR method based on size-differences in the length of the serum albumin intron-1 and the RanaCR1, a non-LTR retrotransposon of the chicken repeat (CR) family. This PCR yields taxon-specific banding patterns that can easily be screened by standard agarose gel electrophoresis and correctly identify species in all of the 160 samples that had been identified to karyotype with other methods. To distinguish ploidy levels in LR, LLR and RRL specimens, we used the ratio of the peak heights of the larger (ridibundus specific) to the smaller (lessonae specific) bands of fluorescently labelled PCR products resolved on a capillary DNA sequencer and obtained a correct assignment of the karyotype in 93% of cases. Our new method will cut down time and expenses drastically for a reliable identification of water frogs of the P. esculentus complex and potentially for identification of other hybridogenetic complexes and/or taxa, and it even serves as a good indicator of the ploidy status of hybrid individuals.

  13. Aerodynamic method for obtaining the soil water retention curve

    NASA Astrophysics Data System (ADS)

    Alekseev, V. V.; Maksimov, I. I.

    2013-07-01

    A new method for the rapid plotting of the soil water retention curve (SWRC) has been proposed that considers the soil water as an environment limited by the soil solid phase on one side and by the soil air on the other side. Both contact surfaces have surface energies, which play the main role in water retention. The use of an idealized soil model with consideration for the nonequilibrium thermodynamic laws and the aerodynamic similarity principles allows us to estimate the volumetric specific surface areas of soils and, using the proposed pedotransfer function (PTF), to plot the SWRC. The volumetric specific surface area of the solid phase, the porosity, and the specific free surface energy at the water-air interface are used as the SWRC parameters. Devices for measuring the parameters are briefly described. The differences between the proposed PTF and the experimental data have been analyzed using the statistical processing of the data.

  14. Method for rapid, high sensitivity tritiated water extraction

    SciTech Connect

    Failor, R.; Belovodsky, L.; Gaevoy, V.; Golubev, A.

    1997-04-20

    We have developed a thermal vacuum desorption process to rapidly extract water from environmental samples for tritium analysis. Thermal vacuum desorption allows for extraction of the moisture from the sample within a few hours in a form and quantity suitable for liquid scintillation counting and allows detection of tritium at the levels of <2 Bq/L of milk, <0.5 Bq/gm of vegetation, and < 0.5 Bq/gin of soil. We developed a prototype unit that can process batches of twenty or more samples within 24 hours. Early data shows that a high percentage of water is extracted reproducibly without enrichment or depletion of the tritium content. The quench coefficient of the extracted water is low allowing for accurate, direct liquid scintillation counting. Excellent comparison has been observed with results using freeze-dry lypholization as the water extraction method.

  15. Method and apparatus for extracting water from air

    DOEpatents

    Spletzer, Barry L.; Callow, Diane Schafer; Marron, Lisa C.; Salton, Jonathan R.

    2002-01-01

    The present invention provides a method and apparatus for extracting liquid water from moist air using minimal energy input. The method comprises compressing moist air under conditions that foster the condensation of liquid water. The air can be decompressed under conditions that do not foster the vaporization of the condensate. The decompressed, dried air can be exchanged for a fresh charge of moist air and the process repeated. The liquid condensate can be removed for use. The apparatus can comprise a compression chamber having a variable internal volume. An intake port allows moist air into the compression chamber. An exhaust port allows dried air out of the compression chamber. A condensation device fosters condensation at the desired conditions. A condensate removal port allows liquid water to be removed.

  16. Method and apparatus for extracting water from air

    DOEpatents

    Spletzer, Barry L.

    2001-01-01

    The present invention provides a method and apparatus for extracting liquid water from moist air using minimal energy input. The method comprises compressing moist air under conditions that foster the condensation of liquid water (ideally isothermal to a humidity of 1.0, then adiabatic thereafter). The air can be decompressed under conditions that do not foster the vaporization of the condensate. The decompressed, dried air can be exchanged for a fresh charge of moist air and the process repeated. The liquid condensate can be removed for use. The apparatus can comprise a compression chamber having a variable internal volume. An intake port allows moist air into the compression chamber. An exhaust port allows dried air out of the compression chamber. A condensation device fosters condensation at the desired conditions. A condensate removal port allows liquid water to be removed.

  17. Geomatic methods at the service of water resources modelling

    NASA Astrophysics Data System (ADS)

    Molina, José-Luis; Rodríguez-Gonzálvez, Pablo; Molina, Mª Carmen; González-Aguilera, Diego; Espejo, Fernando

    2014-02-01

    Acquisition, management and/or use of spatial information are crucial for the quality of water resources studies. In this sense, several geomatic methods arise at the service of water modelling, aiming the generation of cartographic products, especially in terms of 3D models and orthophotos. They may also perform as tools for problem solving and decision making. However, choosing the right geomatic method is still a challenge in this field. That is mostly due to the complexity of the different applications and variables involved for water resources management. This study is aimed to provide a guide to best practices in this context by tackling a deep review of geomatic methods and their suitability assessment for the following study types: Surface Hydrology, Groundwater Hydrology, Hydraulics, Agronomy, Morphodynamics and Geotechnical Processes. This assessment is driven by several decision variables grouped in two categories, classified depending on their nature as geometric or radiometric. As a result, the reader comes with the best choice/choices for the method to use, depending on the type of water resources modelling study in hand.

  18. Screening Methods for Metal-Containing Nanoparticles in Water

    EPA Science Inventory

    Screening-level analysis of water for metal-containing nanoparticles is achieved with single particle-inductively coupled plasma mass spectrometry (SP-ICPMS). This method measures both the concentration of nanoparticles containing an analyte metal and the mass of the metal in eac...

  19. Determination of the Electronics Charge--Electrolysis of Water Method.

    ERIC Educational Resources Information Center

    Venkatachar, Arun C.

    1985-01-01

    Presents an alternative method for measuring the electronic charge using data from the electrolysis of acidified distilled water. The process (carried out in a commercially available electrolytic cell) has the advantage of short completion time so that students can determine electron charge and mass in one laboratory period. (DH)

  20. Comparison of commonly used procedures, including the doubly-labelled water technique, in the estimation of total energy expenditure of women with special reference to the significance of body fatness.

    PubMed

    Lof, Marie; Hannestad, Ulf; Forsum, Elisabet

    2003-11-01

    According to the report of the World Health Organization (1985), total energy expenditure (TEE) in human subjects can be calculated as BMR x physical activity level (PAL). However, other reports have pointed out limitations in the suggested procedure related to the % body fat of the subjects. The purpose of the present study was to evaluate the World Health Organization (1985) procedure in thirty-four healthy women with BMI 18-39 kg/m2. BMR and TEE were measured using indirect calorimetry (BMRmeas) and the doubly-labelled water method (TEEref) respectively. When assessed using the doubly-labelled water and skinfold-thickness methods, the women had 34 (SD 8) and 33 (SD 6) % body fat respectively. On the basis of guidelines provided by the World Health Organization (1985), 1.64 was selected to represent the average PAL of the women. Furthermore, PAL was also assessed by means of an accelerometer (PALacc), heart-rate recordings (PAL(HR)) and a questionnaire (PALq). These estimates were: PALacc 1.71 (SD 0.17), PAL(HR) 1.76 (SD 0.24), PALq 1.86 (SD 0.27). These values were lower than TEEref/BMRref, which was 1.98 (SD 0.21). BMR assessed using equations recommended by the World Health Organization (1985) (BMRpredicted) overestimated BMR by 594 (SD 431) kJ/24 h. However, when TEE was calculated as BMRpredicted x PALacc, BMRpredicted x PAL(HR) and BMRpredicted x PALq respectively, average results were in agreement with TEEref. Furthermore, TEE values based on BMRpredicted and PALacc, PAL(HR), PALq as well as on PAL = 1.64, minus TEEref, were significantly correlated with body fatness. When the same PAL value (1.64) was used for all subjects, this correlation was particularly strong. Thus, the World Health Organization (1985) procedure may give TEE results that are biased with respect to the body fatness of subjects.

  1. Principles of protein labeling techniques.

    PubMed

    Obermaier, Christian; Griebel, Anja; Westermeier, Reiner

    2015-01-01

    Protein labeling methods prior to separation and analysis have become indispensable approaches for proteomic profiling. Basically, three different types of tags are employed: stable isotopes, mass tags, and fluorophores. While proteins labeled with stable isotopes and mass tags are measured and differentiated by mass spectrometry, fluorescent labels are detected with fluorescence imagers. The major purposes for protein labeling are monitoring of biological processes, reliable quantification of compounds and specific detection of protein modifications and isoforms in multiplexed samples, enhancement of detection sensitivity, and simplification of detection workflows. Proteins can be labeled during cell growth by incorporation of amino acids containing different isotopes, or in biological fluids, cells or tissue samples by attaching specific groups to the ε-amino group of lysine, the N-terminus, or the cysteine residues. The principles and the modifications of the different labeling approaches on the protein level are described; benefits and shortcomings of the methods are discussed.

  2. Development of Fluorescence Label and Con-focal Laser Scanning Microscopy Method for Non-Destructive Local Impurity Distribution Analysis in Protein Crystals

    NASA Astrophysics Data System (ADS)

    Iimura, Yoshikazu; Yoshizaki, Izumi; Nakamura, Hirohiko; Yoda, Shinichi; Komatsu, Hiroshi

    2003-09-01

    A new method for quantitative analysis of the impurity concentration in protein crystals and solutions was developed. This technique utilizes fluorescence label (FL) with con-focal laser scanning microscopy (CLSM), which is more effective than SDS-PAGE analysis currently used for this purpose. The advantages of CLSM are that, it is non-destructive so that the impurity incorporation and local distribution could be observed in situ, and also that only a micro-quantity of protein solution is needed. The impurity protein is labeled with fluorescence material, and mixed with the crystallization solution. The solution and the crystal are observed by CLSM, and the fluorescence intensity from the labeled impurity is then converted to the impurity concentration by using calibration curves. A case study using Hen Egg White Lysozyme as a sample is reported. Calibration curves were obtained by comparing the fluorescence intensity and the actual impurity concentration determined by the absorbance at 280 nm and SDS-PAGE. A few factors such as the numerical aperture of the objective lens or the pinhole size were fixed. The utilization of this technique leads to the understanding of the effect of impurities on protein crystal growth.

  3. Differential quantitative proteomics of Porphyromonas gingivalis by linear ion trap mass spectrometry: Non-label methods comparison, q-values and LOWESS curve fitting

    NASA Astrophysics Data System (ADS)

    Xia, Qiangwei; Wang, Tiansong; Park, Yoonsuk; Lamont, Richard J.; Hackett, Murray

    2007-01-01

    Differential analysis of whole cell proteomes by mass spectrometry has largely been applied using various forms of stable isotope labeling. While metabolic stable isotope labeling has been the method of choice, it is often not possible to apply such an approach. Four different label free ways of calculating expression ratios in a classic "two-state" experiment are compared: signal intensity at the peptide level, signal intensity at the protein level, spectral counting at the peptide level, and spectral counting at the protein level. The quantitative data were mined from a dataset of 1245 qualitatively identified proteins, about 56% of the protein encoding open reading frames from Porphyromonas gingivalis, a Gram-negative intracellular pathogen being studied under extracellular and intracellular conditions. Two different control populations were compared against P. gingivalis internalized within a model human target cell line. The q-value statistic, a measure of false discovery rate previously applied to transcription microarrays, was applied to proteomics data. For spectral counting, the most logically consistent estimate of random error came from applying the locally weighted scatter plot smoothing procedure (LOWESS) to the most extreme ratios generated from a control technical replicate, thus setting upper and lower bounds for the region of experimentally observed random error.

  4. Method for Water Management Considering Long-term Probabilistic Forecasts

    NASA Astrophysics Data System (ADS)

    Hwang, J.; Kang, J.; Suh, A. S.

    2015-12-01

    This research is aimed at predicting the monthly inflow of the Andong-dam basin in South Korea using long-term probabilistic forecasts to apply long-term forecasts to water management. Forecasted Cumulative Distribution Functions (CDFs) of monthly precipitation are plotted by combining the range of monthly precipitation based on proper Probability Density Function (PDF) in past data with probabilistic forecasts in each category. Ensembles of inflow are estimated by entering generated ensembles of precipitation based on the CDFs into the 'abcd' water budget model. The bias and RMSE between averages in past data and observed inflow are compared to them in forecasted ensembles. In our results, the bias and RMSE of average precipitation in the forecasted ensemble are bigger than in past data, whereas the average inflow in the forecasted ensemble is smaller than in past data. This result could be used for reference data to apply long-term forecasts to water management, because of the limit in the number of forecasted data for verification and differences between the Andong-dam basin and the forecasted regions. This research has significance by suggesting a method of applying probabilistic information in climate variables from long-term forecasts to water management in Korea. Original data of a climate model, which produces long-term probabilistic forecasts should be verified directly as input data of a water budget model in the future, so that a more scientific response in water management against uncertainty of climate change could be reached.

  5. Scatterscore : A reconnaissance method to evaluate changes in water quality

    SciTech Connect

    Kim, A.G.; Cardone, C.R.

    2005-12-01

    Water quality data collected in periodic monitoring programs are often difficult to evaluate, especially if the number of parameters is large, the sampling schedule varies, and values are of different orders of magnitude. The Scatterscore Water Quality Evaluation was developed to yield a quantitative score, based on all measured variables in periodic water quality reports, indicating positive, negative or random change. This new methodology calculates a reconnaissance score based on the differences between up-gradient (control) versus down-gradient (treatment) water quality data sets. All parameters measured over a period of time at two or more sampling points are compared. The relationship between the ranges of measured values and the ratio of the medians for each parameter produces a data point that falls into one of four sections on a scattergram. The number and average values of positive, negative and random change points is used to calculate a Scatterscore that indicates the magnitude and direction of overall change in water quality. The Scatterscore Water Quality Evaluation, a reconnaissance method to track general changes, has been applied to 20 sites at which coal utilization by-products (CUB) were used to control acid mine drainage (AMD).

  6. Development of a Rapid Assimilable Organic Carbon Method for Water

    PubMed Central

    LeChevallier, Mark W.; Shaw, Nancy E.; Kaplan, Louis A.; Bott, Thomas L.

    1993-01-01

    A rapid method for measurement of assimilable organic carbon (AOC) is proposed. The time needed to perform the assay is reduced by increasing the incubation temperature and increasing the inoculum density. The ATP luciferin-luciferase method quickly enumerates the test organisms without the need for plate count media or dilution bottles. There was no significant difference between AOC values determined with strain P17 for the ATP and plate count procedures. For strain NOX, the plate count procedure underestimated bacterial levels in some samples. Comparison of AOC values obtained by the Belleville laboratory (by the ATP technique) and the Stroud Water Research Center (by plate counts) showed that values were significantly correlated and not significantly different. The study concludes that the rapid AOC method can quickly determine the bacterial growth potential of water within 2 to 4 days. PMID:16348936

  7. Predicting recreational water quality advisories: A comparison of statistical methods

    USGS Publications Warehouse

    Brooks, Wesley R.; Corsi, Steven R.; Fienen, Michael N.; Carvin, Rebecca B.

    2016-01-01

    Epidemiological studies indicate that fecal indicator bacteria (FIB) in beach water are associated with illnesses among people having contact with the water. In order to mitigate public health impacts, many beaches are posted with an advisory when the concentration of FIB exceeds a beach action value. The most commonly used method of measuring FIB concentration takes 18–24 h before returning a result. In order to avoid the 24 h lag, it has become common to ”nowcast” the FIB concentration using statistical regressions on environmental surrogate variables. Most commonly, nowcast models are estimated using ordinary least squares regression, but other regression methods from the statistical and machine learning literature are sometimes used. This study compares 14 regression methods across 7 Wisconsin beaches to identify which consistently produces the most accurate predictions. A random forest model is identified as the most accurate, followed by multiple regression fit using the adaptive LASSO.

  8. Method to estimate water storage capacity of capillary barriers - Discussion

    SciTech Connect

    Gee, Glendon W. ); Ward, Anderson L. ); Meyer, Philip D. )

    1998-11-01

    This is a brief comment on a previously published paper. The paper by Stormont and Morris[JGGE 124 (4):297-302] provides an interesting approach to computing water storage capacity of capillary barriers used as landfill covers. They correctly show that available water storage capacity can be increased up to a factor of two for a silt loam soil, when it is used in a capillary barrier as compared to existing as a deep soil profile. For this very reason such a capillary barrier, utilizing silt loam soil, was constructed and successfully tested at the U. S. Department of Energy?s Hanford Site in southeastern Washington State. Silt loam soil provides optimal water storage for capillary barriers and ensures minimal drainage. Less benefits are obtained when capillary barriers utilize more sandy soils. We would endorse a limited application of the method of Stormont and Morris. We suggest that there will be large uncertainties in field capacity, wilting point and water retention characteristics and only when these uncertainties are accounted for can such a method be used to provide sound engineering judgement for cover design. A recommended procedure for using this method would include actual field measurements of the soil hydraulic properties of the cover materials.

  9. Rapid quantification method for Legionella pneumophila in surface water.

    PubMed

    Wunderlich, Anika; Torggler, Carmen; Elsässer, Dennis; Lück, Christian; Niessner, Reinhard; Seidel, Michael

    2016-03-01

    World-wide legionellosis outbreaks caused by evaporative cooling systems have shown that there is a need for rapid screening methods for Legionella pneumophila in water. Antibody-based methods for the quantification of L. pneumophila are rapid, non-laborious, and relatively cheap but not sensitive enough for establishment as a screening method for surface and drinking water. Therefore, preconcentration methods have to be applied in advance to reach the needed sensitivity. In a basic test, monolithic adsorption filtration (MAF) was used as primary preconcentration method that adsorbs L. pneumophila with high efficiency. Ten-liter water samples were concentrated in 10 min and further reduced to 1 mL by centrifugal ultrafiltration (CeUF). The quantification of L. pneumophila strains belonging to the monoclonal subtype Bellingham was performed via flow-based chemiluminescence sandwich microarray immunoassays (CL-SMIA) in 36 min. The whole analysis process takes 90 min. A polyclonal antibody (pAb) against L. pneumophila serogroup 1-12 and a monoclonal antibody (mAb) against L. pneumophila SG 1 strain Bellingham were immobilized on a microarray chip. Without preconcentration, the detection limit was 4.0 × 10(3) and 2.8 × 10(3) CFU/mL determined by pAb and mAb 10/6, respectively. For samples processed by MAF-CeUF prior to SMIA detection, the limit of detection (LOD) could be decreased to 8.7 CFU/mL and 0.39 CFU/mL, respectively. A recovery of 99.8 ± 15.9% was achieved for concentrations between 1-1000 CFU/mL. The established combined analytical method is sensitive for rapid screening of surface and drinking water to allow fast hygiene control of L. pneumophila. PMID:26873217

  10. Rapid quantification method for Legionella pneumophila in surface water.

    PubMed

    Wunderlich, Anika; Torggler, Carmen; Elsässer, Dennis; Lück, Christian; Niessner, Reinhard; Seidel, Michael

    2016-03-01

    World-wide legionellosis outbreaks caused by evaporative cooling systems have shown that there is a need for rapid screening methods for Legionella pneumophila in water. Antibody-based methods for the quantification of L. pneumophila are rapid, non-laborious, and relatively cheap but not sensitive enough for establishment as a screening method for surface and drinking water. Therefore, preconcentration methods have to be applied in advance to reach the needed sensitivity. In a basic test, monolithic adsorption filtration (MAF) was used as primary preconcentration method that adsorbs L. pneumophila with high efficiency. Ten-liter water samples were concentrated in 10 min and further reduced to 1 mL by centrifugal ultrafiltration (CeUF). The quantification of L. pneumophila strains belonging to the monoclonal subtype Bellingham was performed via flow-based chemiluminescence sandwich microarray immunoassays (CL-SMIA) in 36 min. The whole analysis process takes 90 min. A polyclonal antibody (pAb) against L. pneumophila serogroup 1-12 and a monoclonal antibody (mAb) against L. pneumophila SG 1 strain Bellingham were immobilized on a microarray chip. Without preconcentration, the detection limit was 4.0 × 10(3) and 2.8 × 10(3) CFU/mL determined by pAb and mAb 10/6, respectively. For samples processed by MAF-CeUF prior to SMIA detection, the limit of detection (LOD) could be decreased to 8.7 CFU/mL and 0.39 CFU/mL, respectively. A recovery of 99.8 ± 15.9% was achieved for concentrations between 1-1000 CFU/mL. The established combined analytical method is sensitive for rapid screening of surface and drinking water to allow fast hygiene control of L. pneumophila.

  11. A new method for water desalination using microbial desalination cells.

    PubMed

    Cao, Xiaoxin; Huang, Xia; Liang, Peng; Xiao, Kang; Zhou, Yingjun; Zhang, Xiaoyuan; Logan, Bruce E

    2009-09-15

    Current water desalination techniques are energy intensive and some use membranes operated at high pressures. It is shown here that water desalination can be accomplished without electrical energy input or high water pressure by using a source of organic matter as the fuel to desalinate water. A microbial fuel cell was modified by placing two membranes between the anode and cathode, creating a middle chamber for water desalination between the membranes. An anion exchange membrane was placed adjacent to the anode, and a cation exchange membrane was positioned next to the cathode. When current was produced by bacteria on the anode, ionic species in the middle chamber were transferred into the two electrode chambers, desalinating the water in the middle chamber. Proof-of-concept experiments for this approach, using what we call a microbial desalination cell (MDC), was demonstrated using water at different initial salt concentrations (5, 20, and 35 g/L) with acetate used as the substrate for the bacteria. The MDC produced a maximum of 2 W/m2 (31 W/m3) while at the same time removing about 90% of the salt in a single desalination cycle. As the salt was removed from the middle chamber the ohmic resistance of the MDC (measured using electrochemical impedance spectroscopy) increased from 25 Omega to 970 Omega at the end of the cycle. This increased resistance was reflected by a continuous decrease in the voltage produced over the cycle. These results demonstrate for the first time the possibility for a new method for water desalination and power production that uses only a source of biodegradable organic matter and bacteria.

  12. Comparison and verification of bacterial water quality indicator measurement methods using ambient coastal water samples.

    PubMed

    Griffith, John F; Aumand, Larissa A; Lee, Ioannice M; McGee, Charles D; Othman, Laila L; Ritter, Kerry J; Walker, Kathy O; Weisberg, Stephen B

    2006-05-01

    More than 30 laboratories routinely monitor water along southern California's beaches for bacterial indicators of fecal contamination. Data from these efforts frequently are combined and compared even though three different methods (membrane filtration (MF), multiple tube fermentation (MTF), and chromogenic substrate (CS) methods) are used. To assess data comparability and quantify variability within method and across laboratories, 26 laboratories participated in an intercalibration exercise. Each laboratory processed three replicates from eight ambient water samples employing the method or methods they routinely use for water quality monitoring. Verification analyses also were conducted on a subset of wells from the CS analysis to confirm or exclude the presence of the target organism. Enterococci results were generally comparable across methods. Confirmation revealed a 9% false positive rate and a 4% false negative rate in the CS method for enterococci, though these errors were small in the context of within- and among-laboratory variability. Fecal coliforms also were comparable across all methods, though CS underestimated the other methods by about 10%, probably because it measures only E. coli, rather than the larger fecal coliform group measured by MF and MTF. CS overestimated total coliforms relative to the other methods by several fold and was found to have a 40% false positive rate in verification. Across-laboratory variability was small relative to within- and among-method variability, but only after data entry errors were corrected. One fifth of the laboratories committed data entry errors that were much larger than any method-related errors. These errors are particularly significant because these data were submitted in a test situation where laboratories were aware they would be under increased scrutiny. Under normal circumstances, it is unlikely that these errors would have been detected and managers would have been obliged to issue beach water quality

  13. Water accessibility in a membrane-inserting peptide comparing Overhauser DNP and pulse EPR methods.

    PubMed

    Segawa, Takuya F; Doppelbauer, Maximilian; Garbuio, Luca; Doll, Andrin; Polyhach, Yevhen O; Jeschke, Gunnar

    2016-05-21

    Water accessibility is a key parameter for the understanding of the structure of biomolecules, especially membrane proteins. Several experimental techniques based on the combination of electron paramagnetic resonance (EPR) spectroscopy with site-directed spin labeling are currently available. Among those, we compare relaxation time measurements and electron spin echo envelope modulation (ESEEM) experiments using pulse EPR with Overhauser dynamic nuclear polarization (DNP) at X-band frequency and a magnetic field of 0.33 T. Overhauser DNP transfers the electron spin polarization to nuclear spins via cross-relaxation. The change in the intensity of the (1)H NMR spectrum of H2O at a Larmor frequency of 14 MHz under a continuous-wave microwave irradiation of the nitroxide spin label contains information on the water accessibility of the labeled site. As a model system for a membrane protein, we use the hydrophobic α-helical peptide WALP23 in unilamellar liposomes of DOPC. Water accessibility measurements with all techniques are conducted for eight peptides with different spin label positions and low radical concentrations (10-20 μM). Consistently in all experiments, the water accessibility appears to be very low, even for labels positioned near the end of the helix. The best profile is obtained by Overhauser DNP, which is the only technique that succeeds in discriminating neighboring positions in WALP23. Since the concentration of the spin-labeled peptides varied, we normalized the DNP parameter ϵ, being the relative change of the NMR intensity, by the electron spin concentration, which was determined from a continuous-wave EPR spectrum.

  14. Water accessibility in a membrane-inserting peptide comparing Overhauser DNP and pulse EPR methods

    NASA Astrophysics Data System (ADS)

    Segawa, Takuya F.; Doppelbauer, Maximilian; Garbuio, Luca; Doll, Andrin; Polyhach, Yevhen O.; Jeschke, Gunnar

    2016-05-01

    Water accessibility is a key parameter for the understanding of the structure of biomolecules, especially membrane proteins. Several experimental techniques based on the combination of electron paramagnetic resonance (EPR) spectroscopy with site-directed spin labeling are currently available. Among those, we compare relaxation time measurements and electron spin echo envelope modulation (ESEEM) experiments using pulse EPR with Overhauser dynamic nuclear polarization (DNP) at X-band frequency and a magnetic field of 0.33 T. Overhauser DNP transfers the electron spin polarization to nuclear spins via cross-relaxation. The change in the intensity of the 1H NMR spectrum of H2O at a Larmor frequency of 14 MHz under a continuous-wave microwave irradiation of the nitroxide spin label contains information on the water accessibility of the labeled site. As a model system for a membrane protein, we use the hydrophobic α-helical peptide WALP23 in unilamellar liposomes of DOPC. Water accessibility measurements with all techniques are conducted for eight peptides with different spin label positions and low radical concentrations (10-20 μM). Consistently in all experiments, the water accessibility appears to be very low, even for labels positioned near the end of the helix. The best profile is obtained by Overhauser DNP, which is the only technique that succeeds in discriminating neighboring positions in WALP23. Since the concentration of the spin-labeled peptides varied, we normalized the DNP parameter ɛ, being the relative change of the NMR intensity, by the electron spin concentration, which was determined from a continuous-wave EPR spectrum.

  15. Rapid synthesis of water-dispersible superparamagnetic iron oxide nanoparticles by a microwave-assisted route for safe labeling of endothelial progenitor cells.

    PubMed

    Carenza, Elisa; Barceló, Verónica; Morancho, Anna; Montaner, Joan; Rosell, Anna; Roig, Anna

    2014-08-01

    We synthesize highly crystalline citrate-coated iron oxide superparamagnetic nanoparticles that are stable and readily dispersible in water by an extremely fast microwave-assisted route and investigate the uptake of magnetic nanoparticles by endothelial cells. Nanoparticles form large aggregates when added to complete endothelial cell medium. The size of the aggregates was controlled by adjusting the ionic strength of the medium. The internalization of nanoparticles into endothelial cells was then investigated by transmission electron microscopy, magnetometry and chemical analysis, together with cell viability assays. Interestingly, a sevenfold more efficient uptake was found for systems with larger nanoparticle aggregates, which also showed significantly higher magnetic resonance imaging effectiveness without compromising cell viability and functionality. We are thus presenting an example of a straightforward microwave synthesis of citrate-coated iron oxide nanoparticles for safe endothelial progenitor cell labeling and good magnetic resonance cell imaging with potential application for magnetic cell guidance and in vivo cell tracking.

  16. Label and Label-Free Detection Techniques for Protein Microarrays

    PubMed Central

    Syahir, Amir; Usui, Kenji; Tomizaki, Kin-ya; Kajikawa, Kotaro; Mihara, Hisakazu

    2015-01-01

    Protein microarray technology has gone through numerous innovative developments in recent decades. In this review, we focus on the development of protein detection methods embedded in the technology. Early microarrays utilized useful chromophores and versatile biochemical techniques dominated by high-throughput illumination. Recently, the realization of label-free techniques has been greatly advanced by the combination of knowledge in material sciences, computational design and nanofabrication. These rapidly advancing techniques aim to provide data without the intervention of label molecules. Here, we present a brief overview of this remarkable innovation from the perspectives of label and label-free techniques in transducing nano-biological events.

  17. A Simple and Efficient In Vivo Non-viral RNA Transfection Method for Labeling the Whole Axonal Tree of Individual Adult Long-Range Projection Neurons.

    PubMed

    Porrero, César; Rodríguez-Moreno, Javier; Quetglas, José I; Smerdou, Cristian; Furuta, Takahiro; Clascá, Francisco

    2016-01-01

    We report a highly efficient, simple, and non-infective method for labeling individual long-range projection neurons (LRPNs) in a specific location with enough sparseness and intensity to allow complete and unambiguous reconstructions of their entire axonal tree. The method is based on the "in vivo" transfection of a large RNA construct that drives the massive expression of green fluorescent protein. The method combines two components: injection of a small volume of a hyperosmolar NaCl solution containing the Pal-eGFP-Sindbis RNA construct (Furuta et al., 2001), followed by the application of high-frequency electric current pulses through the micropipette tip. We show that, although each component alone increases transfection efficacy, compared to simple volume injections of standard RNA solution, the highest efficacy (85.7%) is achieved by the combination of both components. In contrast with the infective viral Sindbis vector, RNA transfection occurs exclusively at the position of the injection micropipette tip. This method simplifies consistently labeling one or a few isolated neurons per brain, a strategy that allows unambiguously resolving and quantifying the brain-wide and often multi-branched monosynaptic circuits created by LRPNs.

  18. A Simple and Efficient In Vivo Non-viral RNA Transfection Method for Labeling the Whole Axonal Tree of Individual Adult Long-Range Projection Neurons

    PubMed Central

    Porrero, César; Rodríguez-Moreno, Javier; Quetglas, José I.; Smerdou, Cristian; Furuta, Takahiro; Clascá, Francisco

    2016-01-01

    We report a highly efficient, simple, and non-infective method for labeling individual long-range projection neurons (LRPNs) in a specific location with enough sparseness and intensity to allow complete and unambiguous reconstructions of their entire axonal tree. The method is based on the “in vivo” transfection of a large RNA construct that drives the massive expression of green fluorescent protein. The method combines two components: injection of a small volume of a hyperosmolar NaCl solution containing the Pal-eGFP-Sindbis RNA construct (Furuta et al., 2001), followed by the application of high-frequency electric current pulses through the micropipette tip. We show that, although each component alone increases transfection efficacy, compared to simple volume injections of standard RNA solution, the highest efficacy (85.7%) is achieved by the combination of both components. In contrast with the infective viral Sindbis vector, RNA transfection occurs exclusively at the position of the injection micropipette tip. This method simplifies consistently labeling one or a few isolated neurons per brain, a strategy that allows unambiguously resolving and quantifying the brain-wide and often multi-branched monosynaptic circuits created by LRPNs. PMID:27047347

  19. A Simple and Efficient In Vivo Non-viral RNA Transfection Method for Labeling the Whole Axonal Tree of Individual Adult Long-Range Projection Neurons.

    PubMed

    Porrero, César; Rodríguez-Moreno, Javier; Quetglas, José I; Smerdou, Cristian; Furuta, Takahiro; Clascá, Francisco

    2016-01-01

    We report a highly efficient, simple, and non-infective method for labeling individual long-range projection neurons (LRPNs) in a specific location with enough sparseness and intensity to allow complete and unambiguous reconstructions of their entire axonal tree. The method is based on the "in vivo" transfection of a large RNA construct that drives the massive expression of green fluorescent protein. The method combines two components: injection of a small volume of a hyperosmolar NaCl solution containing the Pal-eGFP-Sindbis RNA construct (Furuta et al., 2001), followed by the application of high-frequency electric current pulses through the micropipette tip. We show that, although each component alone increases transfection efficacy, compared to simple volume injections of standard RNA solution, the highest efficacy (85.7%) is achieved by the combination of both components. In contrast with the infective viral Sindbis vector, RNA transfection occurs exclusively at the position of the injection micropipette tip. This method simplifies consistently labeling one or a few isolated neurons per brain, a strategy that allows unambiguously resolving and quantifying the brain-wide and often multi-branched monosynaptic circuits created by LRPNs. PMID:27047347

  20. Polyoxometalate water oxidation catalysts and methods of use thereof

    DOEpatents

    Hill, Craig L.; Gueletii, Yurii V.; Musaev, Djamaladdin G.; Yin, Qiushi; Botar, Bogdan

    2014-09-02

    Homogeneous water oxidation catalysts (WOCs) for the oxidation of water to produce hydrogen ions and oxygen, and methods of making and using thereof are described herein. In a preferred embodiment, the WOC is a polyoxometalate WOC which is hydrolytically stable, oxidatively stable, and thermally stable. The WOC oxidized waters in the presence of an oxidant. The oxidant can be generated photochemically, using light, such as sunlight, or electrochemically using a positively biased electrode. The hydrogen ions are subsequently reduced to form hydrogen gas, for example, using a hydrogen evolution catalyst (HEC). The hydrogen gas can be used as a fuel in combustion reactions and/or in hydrogen fuel cells. The catalysts described herein exhibit higher turn over numbers, faster turn over frequencies, and/or higher oxygen yields than prior art catalysts.

  1. Thermal Methods for Investigating Ground-Water Recharge

    USGS Publications Warehouse

    Blasch, Kyle W.; Constantz, Jim; Stonestrom, David A.

    2007-01-01

    Recharge of aquifers within arid and semiarid environments is defined as the downward flux of water across the regional water table. The introduction of recharging water at the land surface can occur at discreet locations, such as in stream channels, or be distributed over the landscape, such as across broad interarroyo areas within an alluvial ground-water basin. The occurrence of recharge at discreet locations is referred to as focused recharge, whereas the occurrence of recharge over broad regions is referred to as diffuse recharge. The primary interest of this appendix is focused recharge, but regardless of the type of recharge, estimation of downward fluxes is essential to its quantification. Like chemical tracers, heat can come from natural sources or be intentionally introduced to infer transport properties and aquifer recharge. The admission and redistribution of heat from natural processes such as insolation, infiltration, and geothermal activity can be used to quantify subsurface flow regimes. Heat is well suited as a ground-water tracer because it provides a naturally present dynamic signal and is relatively harmless over a useful range of induced perturbations. Thermal methods have proven valuable for recharge investigations for several reasons. First, theoretical descriptions of coupled water-and-heat transport are available for the hydrologic processes most often encountered in practice. These include land-surface mechanisms such as radiant heating from the sun, radiant cooling into space, and evapotranspiration, in addition to the advective and conductive mechanisms that usually dominate at depth. Second, temperature is theoretically well defined and readily measured. Third, thermal methods for depths ranging from the land surface to depths of hundreds of meters are based on similar physical principles. Fourth, numerical codes for simulating heat and water transport have become increasingly reliable and widely available. Direct measurement of water

  2. Geophysical Methods for Investigating Ground-Water Recharge

    USGS Publications Warehouse

    Ferre, Ty P.A.; Binley, Andrew M.; Blasch, Kyle W.; Callegary, James B.; Crawford, Steven M.; Fink, James B.; Flint, Alan L.; Flint, Lorraine E.; Hoffmann, John P.; Izbicki, John A.; Levitt, Marc T.; Pool, Donald R.; Scanlon, Bridget R.

    2007-01-01

    While numerical modeling has revolutionized our understanding of basin-scale hydrologic processes, such models rely almost exclusively on traditional measurements?rainfall, streamflow, and water-table elevations?for calibration and testing. Model calibration provides initial estimates of ground-water recharge. Calibrated models are important yet crude tools for addressing questions about the spatial and temporal distribution of recharge. An inverse approach to recharge estimation is taken of necessity, due to inherent difficulties in making direct measurements of flow across the water table. Difficulties arise because recharging fluxes are typically small, even in humid regions, and because the location of the water table changes with time. Deep water tables in arid and semiarid regions make recharge monitoring especially difficult. Nevertheless, recharge monitoring must advance in order to improve assessments of ground-water recharge. Improved characterization of basin-scale recharge is critical for informed water-resources management. Difficulties in directly measuring recharge have prompted many efforts to develop indirect methods. The mass-balance approach of estimating recharge as the residual of generally much larger terms has persisted despite the use of increasing complex and finely gridded large-scale hydrologic models. Geophysical data pertaining to recharge rates, timing, and patterns have the potential to substantially improve modeling efforts by providing information on boundary conditions, by constraining model inputs, by testing simplifying assumptions, and by identifying the spatial and temporal resolutions needed to predict recharge to a specified tolerance in space and in time. Moreover, under certain conditions, geophysical measurements can yield direct estimates of recharge rates or changes in water storage, largely eliminating the need for indirect measures of recharge. This appendix presents an overview of physically based, geophysical methods

  3. 13C-labeled gluconate tracing as a direct and accurate method for determining the pentose phosphate pathway split ratio in Penicillium chrysogenum.

    PubMed

    Kleijn, Roelco J; van Winden, Wouter A; Ras, Cor; van Gulik, Walter M; Schipper, Dick; Heijnen, Joseph J

    2006-07-01

    In this study we developed a new method for accurately determining the pentose phosphate pathway (PPP) split ratio, an important metabolic parameter in the primary metabolism of a cell. This method is based on simultaneous feeding of unlabeled glucose and trace amounts of [U-13C]gluconate, followed by measurement of the mass isotopomers of the intracellular metabolites surrounding the 6-phosphogluconate node. The gluconate tracer method was used with a penicillin G-producing chemostat culture of the filamentous fungus Penicillium chrysogenum. For comparison, a 13C-labeling-based metabolic flux analysis (MFA) was performed for glycolysis and the PPP of P. chrysogenum. For the first time mass isotopomer measurements of 13C-labeled primary metabolites are reported for P. chrysogenum and used for a 13C-based MFA. Estimation of the PPP split ratio of P. chrysogenum at a growth rate of 0.02 h(-1) yielded comparable values for the gluconate tracer method and the 13C-based MFA method, 51.8% and 51.1%, respectively. A sensitivity analysis of the estimated PPP split ratios showed that the 95% confidence interval was almost threefold smaller for the gluconate tracer method than for the 13C-based MFA method (40.0 to 63.5% and 46.0 to 56.5%, respectively). From these results we concluded that the gluconate tracer method permits accurate determination of the PPP split ratio but provides no information about the remaining cellular metabolism, while the 13C-based MFA method permits estimation of multiple fluxes but provides a less accurate estimate of the PPP split ratio.

  4. Treatment methods for breaking certain oil and water emulsions

    DOEpatents

    Sealock, Jr., L. John; Baker, Eddie G.; Elliott, Douglas C.

    1992-01-01

    Disclosed are treatment methods for breaking emulsions of petroleum oil and salt water, fatty oil and water, and those resulting from liquefication of organic material. The emulsions are broken by heating to a predetermined temperature at or above about 200.degree. C. and pressurizing to a predetermined pressure above the vapor pressure of water at the predetermined temperature to produce a heated and pressurized fluid. The heated and pressurized fluid is contained in a single vessel at the predetermined temperature and pressure for a predetermined period of time to effectively separate the emulsion into substantially distinct first and second phases, the first phase comprising primarily the petroleum oil, the second phase comprising primarily the water. The first and second phases are separately withdrawn from the vessel at a withdraw temperature between about 200.degree. C. and 374.degree. C. and a withdraw pressure above the vapor pressure of water at the withdraw temperature. Where solids are present in the certain emulsions, the above described treatment may also effectively separate the certain emulsion into a substantially distinct third phase comprising primarily the solids.

  5. Simulating Space Capsule Water Landing with Explicit Finite Element Method

    NASA Technical Reports Server (NTRS)

    Wang, John T.; Lyle, Karen H.

    2007-01-01

    A study of using an explicit nonlinear dynamic finite element code for simulating the water landing of a space capsule was performed. The finite element model contains Lagrangian shell elements for the space capsule and Eulerian solid elements for the water and air. An Arbitrary Lagrangian Eulerian (ALE) solver and a penalty coupling method were used for predicting the fluid and structure interaction forces. The space capsule was first assumed to be rigid, so the numerical results could be correlated with closed form solutions. The water and air meshes were continuously refined until the solution was converged. The converged maximum deceleration predicted is bounded by the classical von Karman and Wagner solutions and is considered to be an adequate solution. The refined water and air meshes were then used in the models for simulating the water landing of a capsule model that has a flexible bottom. For small pitch angle cases, the maximum deceleration from the flexible capsule model was found to be significantly greater than the maximum deceleration obtained from the corresponding rigid model. For large pitch angle cases, the difference between the maximum deceleration of the flexible model and that of its corresponding rigid model is smaller. Test data of Apollo space capsules with a flexible heat shield qualitatively support the findings presented in this paper.

  6. A label-free fluorescent assay for free chlorine in drinking water based on protein-stabilized gold nanoclusters.

    PubMed

    Xiong, Xiaoli; Tang, Yan; Zhang, Liangliang; Zhao, Shulin

    2015-01-01

    Bovine serum albumin stabilized Au nanoclusters (BSA-AuNCs) were demonstrated as a novel fluorescence probe for sensitive and selective detection of free chlorine in drinking water. The fluorescence of BSA-AuNCs was found to be quenched effectively by the free chlorine, and the decrease in fluorescence intensity of BSA-AuNCs allowed the sensitive detection of free chlorine in the range of 0.8-800 μM. The detection limit is 0.50 μM at a signal-to-noise ratio of 3. The present fluorescent assay for free chlorine possesses low detection limit, wide linear range and good selectivity. Real tap water samples were analyzed with satisfactory results, which suggested its potential for water quality analysis.

  7. Pharmacokinetic profile of rizatriptan 10-mg tablet and 10-mg orally disintegrating tablet administered with or without water in healthy subjects: an open-label, randomized, single-dose, 3-period crossover study.

    PubMed

    Swan, Suzanne K; Alcorn, Harry; Rodgers, Anthony; Hustad, Carolyn M; Ramsey, Karen E; Woll, Susan; Skobieranda, Franck

    2006-02-01

    This open-label, 3-period crossover study compared the plasma concentration profiles of rizatriptan tablet, orally disintegrating tablet with water (ODTc), and ODT without water (ODTs) in 24 healthy volunteers aged 18 to 45 years. At each period, subjects received a single dose of either 10-mg rizatriptan tablet, 10-mg rizatriptan ODTs, or 10-mg rizatriptan ODTc. The authors hypothesized that ODTc has a greater geometric mean AUC(0-2h) than ODTs and that ODTc has a greater geometric mean AUC(0-1h) than tablet. A secondary end point was to compare the time of occurrence of the maximum rizatriptan plasma concentration (t(max)) of each dosing method. ODTc had a statistically significantly greater geometric mean AUC(0-2h) compared with ODTs (33.84 h x ng/mL vs 18.83 h x ng/mL; P < .001). ODTc had a slightly, but not statistically significantly, greater geometric mean AUC(0-1h) compared with rizatriptan tablet (17.07 h x ng/mL vs 13.32 h x ng/mL). The median t(max) was 0.67 hours for ODTc and tablet and 1.33 hours for ODTs. ODTc showed a slightly, but not significantly, faster rate of absorption compared with tablet. ODTs with water had a faster rate of absorption than ODTc. Future studies are needed to determine whether this pharmacokinetic difference produces differential efficacy in a clinical setting. PMID:16432269

  8. An automated dynamic water vapor permeation test method

    NASA Astrophysics Data System (ADS)

    Gibson, Phillip; Kendrick, Cyrus; Rivin, Donald; Charmchii, Majid; Sicuranza, Linda

    1995-05-01

    This report describes an automated apparatus developed to measure the transport of water vapor through materials under a variety of conditions. The apparatus is more convenient to use than the traditional test methods for textiles and clothing materials, and allows one to use a wider variety of test conditions to investigate the concentration-dependent and nonlinear transport behavior of many of the semipermeable membrane laminates which are now available. The dynamic moisture permeation cell (DMPC) has been automated to permit multiple setpoint testing under computer control, and to facilitate investigation of transient phenomena. Results generated with the DMPC are in agreement with and of comparable accuracy to those from the ISO 11092 (sweating guarded hot plate) method of measuring water vapor permeability.

  9. Enzyme-labeled Antigen Method: Development and Application of the Novel Approach for Identifying Plasma Cells Locally Producing Disease-specific Antibodies in Inflammatory Lesions

    PubMed Central

    Mizutani, Yasuyoshi; Shiogama, Kazuya; Onouchi, Takanori; Sakurai, Kouhei; Inada, Ken-ichi; Tsutsumi, Yutaka

    2016-01-01

    In chronic inflammatory lesions of autoimmune and infectious diseases, plasma cells are frequently observed. Antigens recognized by antibodies produced by the plasma cells mostly remain unclear. A new technique identifying these corresponding antigens may give us a breakthrough for understanding the disease from a pathophysiological viewpoint, simply because the immunocytes are seen within the lesion. We have developed an enzyme-labeled antigen method for microscopic identification of the antigen recognized by specific antibodies locally produced in plasma cells in inflammatory lesions. Firstly, target biotinylated antigens were constructed by the wheat germ cell-free protein synthesis system or through chemical biotinylation. Next, proteins reactive to antibodies in tissue extracts were screened and antibody titers were evaluated by the AlphaScreen method. Finally, with the enzyme-labeled antigen method using the biotinylated antigens as probes, plasma cells producing specific antibodies were microscopically localized in fixed frozen sections. Our novel approach visualized tissue plasma cells that produced 1) autoantibodies in rheumatoid arthritis, 2) antibodies against major antigens of Porphyromonas gingivalis in periodontitis or radicular cyst, and 3) antibodies against a carbohydrate antigen, Strep A, of Streptococcus pyogenes in recurrent tonsillitis. Evaluation of local specific antibody responses expectedly contributes to clarifying previously unknown processes in inflammatory disorders. PMID:27006517

  10. A new tritiated water measurement method with plastic scintillator pellets.

    PubMed

    Furuta, Etsuko; Iwasaki, Noriko; Kato, Yuka; Tomozoe, Yusuke

    2016-01-01

    A new tritiated water measurement method with plastic scintillator pellets (PS-pellets) by using a conventional liquid scintillation counter was developed. The PS-pellets used were 3 mm in both diameter and length. A low potassium glass vial was filled full with the pellets, and tritiated water was applied to the vial from 5 to 100 μl. Then, the sample solution was scattered in the interstices of the pellets in a vial. This method needs no liquid scintillator, so no liquid organic waste fluid is generated. The counting efficiency with the pellets was approximately 48 % when a 5 μl solution was used, which was higher than that of conventional measurement using liquid scintillator. The relationship between count rate and activity showed good linearity. The pellets were able to be used repeatedly, so few solid wastes are generated with this method. The PS-pellets are useful for tritiated water measurement; however, it is necessary to develop a new device which can be applied to a larger volume and measure low level concentration like an environmental application. PMID:26856930

  11. Cartesian Methods for the Shallow Water Equations on a Sphere

    SciTech Connect

    Drake, J.B.

    2000-02-14

    The shallow water equations in a spherical geometry are solved using a 3-dimensional Cartesian method. Spatial discretization of the 2-dimensional, horizontal differential operators is based on the Cartesian form of the spherical harmonics and an icosahedral (spherical) grid. Computational velocities are expressed in Cartesian coordinates so that a problem with a singularity at the pole is avoided. Solution of auxiliary elliptic equations is also not necessary. A comparison is made between the standard form of the Cartesian equations and a rotational form using a standard set of test problems. Error measures and conservation properties of the method are reported for the test problems.

  12. Evaluation of different field methods for measuring soil water infiltration

    NASA Astrophysics Data System (ADS)

    Pla-Sentís, Ildefonso; Fonseca, Francisco

    2010-05-01

    Soil infiltrability, together with rainfall characteristics, is the most important hydrological parameter for the evaluation and diagnosis of the soil water balance and soil moisture regime. Those balances and regimes are the main regulating factors of the on site water supply to plants and other soil organisms and of other important processes like runoff, surface and mass erosion, drainage, etc, affecting sedimentation, flooding, soil and water pollution, water supply for different purposes (population, agriculture, industries, hydroelectricity), etc. Therefore the direct measurement of water infiltration rates or its indirect deduction from other soil characteristics or properties has become indispensable for the evaluation and modelling of the previously mentioned processes. Indirect deductions from other soil characteristics measured under laboratory conditions in the same soils, or in other soils, through the so called "pedo-transfer" functions, have demonstrated to be of limited value in most of the cases. Direct "in situ" field evaluations have to be preferred in any case. In this contribution we present the results of past experiences in the measurement of soil water infiltration rates in many different soils and land conditions, and their use for deducing soil water balances under variable climates. There are also presented and discussed recent results obtained in comparing different methods, using double and single ring infiltrometers, rainfall simulators, and disc permeameters, of different sizes, in soils with very contrasting surface and profile characteristics and conditions, including stony soils and very sloping lands. It is concluded that there are not methods universally applicable to any soil and land condition, and that in many cases the results are significantly influenced by the way we use a particular method or instrument, and by the alterations in the soil conditions by the land management, but also due to the manipulation of the surface

  13. Application of an enzyme-labeled antigen method for visualizing plasma cells producing antibodies against Strep A, a carbohydrate antigen of Streptococcus pyogenes, in recurrent tonsillitis.

    PubMed

    Onouchi, Takanori; Mizutani, Yasuyoshi; Shiogama, Kazuya; Inada, Ken-ichi; Okada, Tatsuyoshi; Naito, Kensei; Tsutsumi, Yutaka

    2015-01-01

    Streptococcus pyogenes is the main causative pathogen of recurrent tonsillitis. Histologically, lesions of recurrent tonsillitis contain numerous plasma cells. Strep A is an antigenic carbohydrate molecule on the cell wall of S. pyogenes. As expected, plasma cells in subjects with recurrent tonsillitis secrete antibodies against Strep A. The enzyme-labeled antigen method is a novel histochemical technique that visualizes specific antibody-producing cells in tissue sections by employing a biotin-labeled antigen as a probe. The purpose of the present study was to visualize plasma cells producing antibodies reactive with Strep A in recurrent tonsillitis. Firstly, the lymph nodes of rats immunized with boiled S. pyogenes were paraformaldehyde-fixed and specific plasma cells localized in frozen sections with biotinylated Strep A. Secondly, an enzyme-labeled antigen method was used on human tonsil surgically removed from 12 patients with recurrent tonsillitis. S. pyogenes genomes were PCR-detected in all 12 specimens. The emm genotypes belonged to emm12 in nine specimens and emm1 in three. Plasma cells producing anti-Strep A antibodies were demonstrated in prefixed frozen sections of rat lymph nodes, 8/12 human specimens from patients with recurrent tonsillitis but not in two control tonsils. In human tonsils, Strep A-reactive plasma cells were observed within the reticular squamous mucosa and just below the mucosa, and the specific antibodies belonged to either IgA or IgG classes. Our technique is effective in visualizing immunocytes producing specific antibodies against the bacterial carbohydrate antigen, and is thus a novel histochemical tool for analyzing immune reactions in infectious disorders.

  14. A label-free differential quantitative mass spectrometry method for the characterization and identification of protein changes during citrus fruit development

    PubMed Central

    2010-01-01

    Background Citrus is one of the most important and widely grown commodity fruit crops. In this study a label-free LC-MS/MS based shot-gun proteomics approach was taken to explore three main stages of citrus fruit development. These approaches were used to identify and evaluate changes occurring in juice sac cells in various metabolic pathways affecting citrus fruit development and quality. Results Protein changes in citrus juice sac cells were identified and quantified using label-free shotgun methodologies. Two alternative methods, differential mass-spectrometry (dMS) and spectral counting (SC) were used to analyze protein changes occurring during earlier and late stages of fruit development. Both methods were compared in order to develop a proteomics workflow that could be used in a non-model plant lacking a sequenced genome. In order to resolve the bioinformatics limitations of EST databases from species that lack a full sequenced genome, we established iCitrus. iCitrus is a comprehensive sequence database created by merging three major sources of sequences (HarvEST:citrus, NCBI/citrus/unigenes, NCBI/citrus/proteins) and improving the annotation of existing unigenes. iCitrus provided a useful bioinformatics tool for the high-throughput identification of citrus proteins. We have identified approximately 1500 citrus proteins expressed in fruit juice sac cells and quantified the changes of their expression during fruit development. Our results showed that both dMS and SC provided significant information on protein changes, with dMS providing a higher accuracy. Conclusion Our data supports the notion of the complementary use of dMS and SC for label-free comparative proteomics, broadening the identification spectrum and strengthening the identification of trends in protein expression changes during the particular processes being compared. PMID:21162737

  15. Assessment of Crop Water Requirement Methods for Annual Agricultural Water Allocation Planning

    NASA Astrophysics Data System (ADS)

    Aghdasi, F.; Sharifi, M. A.; van der Tol, C.

    2010-05-01

    The potential use of remote sensing in water resource and in particular in irrigation management has been widely acknowledged. However, in reality, operational applications of remote sensing in irrigation management are few. In this study, the applicability of the main available remote sensing based techniques of irrigation management is evaluated in a pilot area in Iran. The evaluated techniques include so called Crop Water Requirement "CWR" methods for the planning of annual water allocation in irrigated agriculture. A total of 40 years of historical weather data were classified into wet, normal, and dry years using a Standardised Precipitation Index (SPI). For each of these three classes the average CWR was calculated. Next, by applying Markov Chain Process to the time series of precipitation, the expected CWR for the forthcoming planning year was estimated. Using proper interpolation techniques the expected CWR at each station was converted to CWR map of the area, which was then used for annual water allocation planning. To estimate the crop water requirement, methods developed for the DEMETER project (DEMonstration of Earth observation Technologies in Routine irrigation advisory services) and Surface Energy Balance System "SEBS" algorithm were used, and their results were compared with conventional methods, including FAO-56 and lysimeter data amongst others. Use was made of both ASTER and MODIS images to determine crop water requirement at local and regional scales. Four methods of estimating crop coefficients were used: DEMETER Kc-NDVI, DEMETER Kc-analytical, FAO-56 and SEBS algorithm. Results showed that DEMETER (analytical approach) and FAO methods with lowest RMSE are more suitable methods for determination of crop coefficient than SEBS, which gives actual rather than potential evapotranspiration. The use of ASTER and MODIS images did not result in significantly different crop coefficients in the pilot area for the DEMETER analytical approach (α=0

  16. A hydrogen gas-water equilibration method produces accurate and precise stable hydrogen isotope ratio measurements in nutrition studies.

    PubMed

    Wong, William W; Clarke, Lucinda L

    2012-11-01

    Stable hydrogen isotope methodology is used in nutrition studies to measure growth, breast milk intake, and energy requirement. Isotope ratio MS is the best instrumentation to measure the stable hydrogen isotope ratios in physiological fluids. Conventional methods to convert physiological fluids to hydrogen gas (H(2)) for mass spectrometric analysis are labor intensive, require special reagent, and involve memory effect and potential isotope fractionation. The objective of this study was to determine the accuracy and precision of a platinum catalyzed H(2)-water equilibration method for stable hydrogen isotope ratio measurements. Time to reach isotopic equilibrium, day-to-day and week-to-week reproducibility, accuracy, and precision of stable hydrogen isotope ratio measurements by the H(2)-water equilibration method were assessed using a Thermo DELTA V Advantage continuous-flow isotope ratio mass spectrometer. It took 3 h to reach isotopic equilibrium. The day-to-day and week-to-week measurements on water and urine samples with natural abundance and enriched levels of deuterium were highly reproducible. The method was accurate to within 2.8 (o)/oo and reproducible to within 4.0 (o)/oo based on analysis of international references. All the outcome variables, whether in urine samples collected in 10 doubly labeled water studies or plasma samples collected in 26 body water studies, did not differ from those obtained using the reference zinc reduction method. The method produced highly accurate estimation on ad libitum energy intakes, body composition, and water turnover rates. The method greatly reduces the analytical cost and could easily be adopted by laboratories equipped with a continuous-flow isotope ratio mass spectrometer.

  17. A Hydrogen Gas-Water Equilibration Method Produces Accurate and Precise Stable Hydrogen Isotope Ratio Measurements in Nutrition Studies12

    PubMed Central

    Wong, William W.; Clarke, Lucinda L.

    2012-01-01

    Stable hydrogen isotope methodology is used in nutrition studies to measure growth, breast milk intake, and energy requirement. Isotope ratio MS is the best instrumentation to measure the stable hydrogen isotope ratios in physiological fluids. Conventional methods to convert physiological fluids to hydrogen gas (H2) for mass spectrometric analysis are labor intensive, require special reagent, and involve memory effect and potential isotope fractionation. The objective of this study was to determine the accuracy and precision of a platinum catalyzed H2-water equilibration method for stable hydrogen isotope ratio measurements. Time to reach isotopic equilibrium, day-to-day and week-to-week reproducibility, accuracy, and precision of stable hydrogen isotope ratio measurements by the H2-water equilibration method were assessed using a Thermo DELTA V Advantage continuous-flow isotope ratio mass spectrometer. It took 3 h to reach isotopic equilibrium. The day-to-day and week-to-week measurements on water and urine samples with natural abundance and enriched levels of deuterium were highly reproducible. The method was accurate to within 2.8 o/oo and reproducible to within 4.0 o/oo based on analysis of international references. All the outcome variables, whether in urine samples collected in 10 doubly labeled water studies or plasma samples collected in 26 body water studies, did not differ from those obtained using the reference zinc reduction method. The method produced highly accurate estimation on ad libitum energy intakes, body composition, and water turnover rates. The method greatly reduces the analytical cost and could easily be adopted by laboratories equipped with a continuous-flow isotope ratio mass spectrometer. PMID:23014490

  18. Synthesis of Isotopically Labeled (13)C3-Simazine and Development of a Simultaneous UPLC-MS/MS Method for the Analysis of Simazine in Soil.

    PubMed

    Song, Yan; Guo, Yangzhen; Zhang, Xia; Yang, Yue; Chen, Shuo; She, Gaimei; She, Dongmei

    2016-01-14

    The isotope dilution mass spectrometry (IDMS) is a highly efficient method for tackling the ion suppression in complex matrix by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), but a lack of commercial internal standards is a limiting factor for these analyses. Herein, an economical and efficient strategy for the synthesis of (13)C3-simazine via a three-step procedure was developed. The isotope-labeled internal standard was used for determination of simazine residue in soil samples. The quantitation method has a limit of detection of 0.015 μg/kg and quantitation of 0.08 μg/kg. The inter-day and intra-day precision of the method were below 4.6%. Recovery values were ranged between 92.9% and 99.2%. All the samples obtained from six provinces in China contained from 1 to 62 μg/kg of simazine.

  19. A Spacecraft Electrical Characteristics Multi-Label Classification Method Based on Off-Line FCM Clustering and On-Line WPSVM.

    PubMed

    Li, Ke; Liu, Yi; Wang, Quanxin; Wu, Yalei; Song, Shimin; Sun, Yi; Liu, Tengchong; Wang, Jun; Li, Yang; Du, Shaoyi

    2015-01-01

    This paper proposes a novel multi-label classification method for resolving the spacecraft electrical characteristics problems which involve many unlabeled test data processing, high-dimensional features, long computing time and identification of slow rate. Firstly, both the fuzzy c-means (FCM) offline clustering and the principal component feature extraction algorithms are applied for the feature selection process. Secondly, the approximate weighted proximal support vector machine (WPSVM) online classification algorithms is used to reduce the feature dimension and further improve the rate of recognition for electrical characteristics spacecraft. Finally, the data capture contribution method by using thresholds is proposed to guarantee the validity and consistency of the data selection. The experimental results indicate that the method proposed can obtain better data features of the spacecraft electrical characteristics, improve the accuracy of identification and shorten the computing time effectively. PMID:26544549

  20. A Spacecraft Electrical Characteristics Multi-Label Classification Method Based on Off-Line FCM Clustering and On-Line WPSVM.

    PubMed

    Li, Ke; Liu, Yi; Wang, Quanxin; Wu, Yalei; Song, Shimin; Sun, Yi; Liu, Tengchong; Wang, Jun; Li, Yang; Du, Shaoyi

    2015-01-01

    This paper proposes a novel multi-label classification method for resolving the spacecraft electrical characteristics problems which involve many unlabeled test data processing, high-dimensional features, long computing time and identification of slow rate. Firstly, both the fuzzy c-means (FCM) offline clustering and the principal component feature extraction algorithms are applied for the feature selection process. Secondly, the approximate weighted proximal support vector machine (WPSVM) online classification algorithms is used to reduce the feature dimension and further improve the rate of recognition for electrical characteristics spacecraft. Finally, the data capture contribution method by using thresholds is proposed to guarantee the validity and consistency of the data selection. The experimental results indicate that the method proposed can obtain better data features of the spacecraft electrical characteristics, improve the accuracy of identification and shorten the computing time effectively.

  1. A Spacecraft Electrical Characteristics Multi-Label Classification Method Based on Off-Line FCM Clustering and On-Line WPSVM

    PubMed Central

    Li, Ke; Liu, Yi; Wang, Quanxin; Wu, Yalei; Song, Shimin; Sun, Yi; Liu, Tengchong; Wang, Jun; Li, Yang; Du, Shaoyi

    2015-01-01

    This paper proposes a novel multi-label classification method for resolving the spacecraft electrical characteristics problems which involve many unlabeled test data processing, high-dimensional features, long computing time and identification of slow rate. Firstly, both the fuzzy c-means (FCM) offline clustering and the principal component feature extraction algorithms are applied for the feature selection process. Secondly, the approximate weighted proximal support vector machine (WPSVM) online classification algorithms is used to reduce the feature dimension and further improve the rate of recognition for electrical characteristics spacecraft. Finally, the data capture contribution method by using thresholds is proposed to guarantee the validity and consistency of the data selection. The experimental results indicate that the method proposed can obtain better data features of the spacecraft electrical characteristics, improve the accuracy of identification and shorten the computing time effectively. PMID:26544549

  2. A lab-on-chip cell-based biosensor for label-free sensing of water toxicants.

    PubMed

    Liu, F; Nordin, A N; Li, F; Voiculescu, I

    2014-04-01

    This paper presents a lab-on-chip biosensor containing an enclosed fluidic cell culturing well seeded with live cells for rapid screening of toxicants in drinking water. The sensor is based on the innovative placement of the working electrode for the electrical cell-substrate impedance sensing (ECIS) technique as the top electrode of a quartz crystal microbalance (QCM) resonator. Cell damage induced by toxic water will cause a decrease in impedance, as well as an increase in the resonant frequency. For water toxicity tests, the biosensor's unique capabilities of performing two complementary measurements simultaneously (impedance and mass-sensing) will increase the accuracy of detection while decreasing the false-positive rate. Bovine aortic endothelial cells (BAECs) were used as toxicity sensing cells. The effects of the toxicants, ammonia, nicotine and aldicarb, on cells were monitored with both the QCM and the ECIS technique. The lab-on-chip was demonstrated to be sensitive to low concentrations of toxicants. The responses of BAECs to toxic samples occurred during the initial 5 to 20 minutes depending on the type of chemical and concentrations. Testing the multiparameter biosensor with aldicarb also demonstrated the hypothesis that using two different sensors to monitor the same cell monolayer provides cross validation and increases the accuracy of detection. For low concentrations of aldicarb, the variations in impedance measurements are insignificant in comparison with the shifts of resonant frequency monitored using the QCM resonator. A highly linear correlation between signal shifts and chemical concentrations was demonstrated for each toxicant.

  3. A lab-on-chip cell-based biosensor for label-free sensing of water toxicants.

    PubMed

    Liu, F; Nordin, A N; Li, F; Voiculescu, I

    2014-04-01

    This paper presents a lab-on-chip biosensor containing an enclosed fluidic cell culturing well seeded with live cells for rapid screening of toxicants in drinking water. The sensor is based on the innovative placement of the working electrode for the electrical cell-substrate impedance sensing (ECIS) technique as the top electrode of a quartz crystal microbalance (QCM) resonator. Cell damage induced by toxic water will cause a decrease in impedance, as well as an increase in the resonant frequency. For water toxicity tests, the biosensor's unique capabilities of performing two complementary measurements simultaneously (impedance and mass-sensing) will increase the accuracy of detection while decreasing the false-positive rate. Bovine aortic endothelial cells (BAECs) were used as toxicity sensing cells. The effects of the toxicants, ammonia, nicotine and aldicarb, on cells were monitored with both the QCM and the ECIS technique. The lab-on-chip was demonstrated to be sensitive to low concentrations of toxicants. The responses of BAECs to toxic samples occurred during the initial 5 to 20 minutes depending on the type of chemical and concentrations. Testing the multiparameter biosensor with aldicarb also demonstrated the hypothesis that using two different sensors to monitor the same cell monolayer provides cross validation and increases the accuracy of detection. For low concentrations of aldicarb, the variations in impedance measurements are insignificant in comparison with the shifts of resonant frequency monitored using the QCM resonator. A highly linear correlation between signal shifts and chemical concentrations was demonstrated for each toxicant. PMID:24463940

  4. Label-Free and Real-Time Monitor of Binding and Dissociation Processes between Protein A and Swine IgG by Oblique-Incidence Reflectivity Difference Method

    NASA Astrophysics Data System (ADS)

    He, Li-Ping; Liu, Shuang; Dai, Jun; Wu, Lin; Liu, Guo-Zhen; Wei, Han-Fu; Lu, Hui-Bin; Jin, Kui-Juan; Yang, Guo-Zhen

    2015-02-01

    Life science has a need for detection methods that are label-free and real-time. In this paper, we have selected staphylococcal protein A (SPA) and swine immunoglobulin G (IgG), and monitor the bindings between SPA and swine IgG with different concentrations, as well as the dissociations of SPA-swine IgG complex in different pH values of phosphate buffer by oblique-incidence reflectivity difference (OIRD) in a label-free and real-time fashion. We obtain the ON and OFF reaction dynamic curves corresponding to the bindings and dissociations of SPA and swine IgG. Through our analysis of the experimental results, we have been able to obtain the damping coefficients and the dissociation time of SPA and swine IgG for different pH values of the phosphate buffer. The results prove that the OIRD technique is a competing method for monitoring the dynamic processes of biomolecule interaction and achieving the quantitative information of reaction kinetics.

  5. Analytical Method for Measuring Cosmogenic (35)S in Natural Waters.

    PubMed

    Urióstegui, Stephanie H; Bibby, Richard K; Esser, Bradley K; Clark, Jordan F

    2015-06-16

    Cosmogenic sulfur-35 in water as dissolved sulfate ((35)SO4) has successfully been used as an intrinsic hydrologic tracer in low-SO4, high-elevation basins. Its application in environmental waters containing high SO4 concentrations has been limited because only small amounts of SO4 can be analyzed using current liquid scintillation counting (LSC) techniques. We present a new analytical method for analyzing large amounts of BaSO4 for (35)S. We quantify efficiency gains when suspending BaSO4 precipitate in Inta-Gel Plus cocktail, purify BaSO4 precipitate to remove dissolved organic matter, mitigate interference of radium-226 and its daughter products by selection of high purity barium chloride, and optimize LSC counting parameters for (35)S determination in larger masses of BaSO4. Using this improved procedure, we achieved counting efficiencies that are comparable to published LSC techniques despite a 10-fold increase in the SO4 sample load. (35)SO4 was successfully measured in high SO4 surface waters and groundwaters containing low ratios of (35)S activity to SO4 mass demonstrating that this new analytical method expands the analytical range of (35)SO4 and broadens the utility of (35)SO4 as an intrinsic tracer in hydrologic settings. PMID:25981756

  6. Development of an LC-MS/MS method for the determination of endogenous cortisol in hair using (13)C3-labeled cortisol as surrogate analyte.

    PubMed

    Binz, Tina M; Braun, Ueli; Baumgartner, Markus R; Kraemer, Thomas

    2016-10-15

    Hair cortisol levels are increasingly applied as a measure for stress in humans and mammals. Cortisol is an endogenous compound and is always present within the hair matrix. Therefore, "cortisol-free hair matrix" is a critical point for any analytical method to accurately quantify especially low cortisol levels. The aim of this project was to modify current methods used for hair cortisol analysis to more accurately determine low endogenous cortisol concentrations in hair. For that purpose, (13)C3-labeled cortisol, which is not naturally present in hair (above 13C natural abundance levels), was used for calibration and comparative validation applying cortisol versus (13)C3-labeled cortisol. Cortisol was extracted from 20mg hair (standard sample amount) applying an optimized single step extraction protocol. An LC-MS/MS method was developed for the quantitative analysis of cortisol using either cortisol or (13)C3-cortisol as calibrators and D7-cortisone as internal standard (IS). The two methods (cortisol/(13)C3-labeled cortisol) were validated in a concentration range up to 500pg/mg and showed good linearity for both analytes (cortisol: R(2)=0.9995; (13)C3-cortisol R(2)=0.9992). Slight differences were observed for limit of detection (LOD) (0.2pg/mg/0.1pg/mg) and limit of quantification (LOQ) (1pg/mg/0.5pg/mg). Precision was good with a maximum deviation of 8.8% and 10% for cortisol and (13)C3-cortisol respectively. Accuracy and matrix effects were good for both analytes except for the quality control (QC) low cortisol. QC low (2.5pg/mg) showed matrix effects (126.5%, RSD 35.5%) and accuracy showed a deviation of 26% when using cortisol to spike. These effects are likely to be caused by the unknown amount of endogenous cortisol in the different hair samples used to determine validation parameters like matrix effect, LOQ and accuracy. No matrix effects were observed for the high QC (400pg/mg) samples. Recovery was good with 92.7%/87.3% (RSD 9.9%/6.2%) for QC low and

  7. A study of the elimination of water from lithium-cationized tripeptide methyl esters by means of tandem mass spectrometry and isotope labeling.

    PubMed

    Talaty, Erach R; Cooper, Travis J; Piland, Debra L; Bateman, David J; Syed, Adeel; Stevenson, William; Van Stipdonk, Michael J

    2006-01-01

    Extensive isotope labeling (2H, 13C and 15N), collision-induced dissociation (CID) and multiple-stage tandem mass spectrometry were used to investigate the elimination of H2O from a series of model, metal-cationized tripeptide methyl esters. The present results corroborate our earlier suggestion that loss of water from lithiated peptides is initiated by a nucleophilic attack from the N-terminal side upon an amide carbonyl carbon atom to form a five-membered ring as an intermediate followed by 1,2-elimination of water. We show that the nucleophilic atom is the oxygen atom of the N-terminal amide group in the fragmentation of [AcGGGOMe+Li]+ as well as [GGGOMe+Li]+. However, the subsequent fragmentation is markedly different in the two cases as a result of the absence and presence of a free amino group. In particular, extensive scrambling of protons in the alpha-positions of GGGOMe is observed, presumably as a consequence of intervention of the basic amino group.

  8. Novel spectrophotometric determination of chloramphenicol and dexamethasone in the presence of non labeled interfering substances using univariate methods and multivariate regression model updating

    NASA Astrophysics Data System (ADS)

    Hegazy, Maha A.; Lotfy, Hayam M.; Rezk, Mamdouh R.; Omran, Yasmin Rostom

    2015-04-01

    Smart and novel spectrophotometric and chemometric methods have been developed and validated for the simultaneous determination of a binary mixture of chloramphenicol (CPL) and dexamethasone sodium phosphate (DSP) in presence of interfering substances without prior separation. The first method depends upon derivative subtraction coupled with constant multiplication. The second one is ratio difference method at optimum wavelengths which were selected after applying derivative transformation method via multiplying by a decoding spectrum in order to cancel the contribution of non labeled interfering substances. The third method relies on partial least squares with regression model updating. They are so simple that they do not require any preliminary separation steps. Accuracy, precision and linearity ranges of these methods were determined. Moreover, specificity was assessed by analyzing synthetic mixtures of both drugs. The proposed methods were successfully applied for analysis of both drugs in their pharmaceutical formulation. The obtained results have been statistically compared to that of an official spectrophotometric method to give a conclusion that there is no significant difference between the proposed methods and the official ones with respect to accuracy and precision.

  9. Novel spectrophotometric determination of chloramphenicol and dexamethasone in the presence of non labeled interfering substances using univariate methods and multivariate regression model updating.

    PubMed

    Hegazy, Maha A; Lotfy, Hayam M; Rezk, Mamdouh R; Omran, Yasmin Rostom

    2015-04-01

    Smart and novel spectrophotometric and chemometric methods have been developed and validated for the simultaneous determination of a binary mixture of chloramphenicol (CPL) and dexamethasone sodium phosphate (DSP) in presence of interfering substances without prior separation. The first method depends upon derivative subtraction coupled with constant multiplication. The second one is ratio difference method at optimum wavelengths which were selected after applying derivative transformation method via multiplying by a decoding spectrum in order to cancel the contribution of non labeled interfering substances. The third method relies on partial least squares with regression model updating. They are so simple that they do not require any preliminary separation steps. Accuracy, precision and linearity ranges of these methods were determined. Moreover, specificity was assessed by analyzing synthetic mixtures of both drugs. The proposed methods were successfully applied for analysis of both drugs in their pharmaceutical formulation. The obtained results have been statistically compared to that of an official spectrophotometric method to give a conclusion that there is no significant difference between the proposed methods and the official ones with respect to accuracy and precision.

  10. Improved biostability assessment of drinking water with a suite of test methods at a water supply treating eutrophic lake water.

    PubMed

    van der Kooij, Dick; Martijn, Bram; Schaap, Peter G; Hoogenboezem, Wim; Veenendaal, Harm R; van der Wielen, Paul W J J

    2015-12-15

    Assessment of drinking-water biostability is generally based on measuring bacterial growth in short-term batch tests. However, microbial growth in the distribution system is affected by multiple interactions between water, biofilms and sediments. Therefore a diversity of test methods was applied to characterize the biostability of drinking water distributed without disinfectant residual at a surface-water supply. This drinking water complied with the standards for the heterotrophic plate count and coliforms, but aeromonads periodically exceeded the regulatory limit (1000 CFU 100 mL(-1)). Compounds promoting growth of the biopolymer-utilizing Flavobacterium johnsoniae strain A3 accounted for c. 21% of the easily assimilable organic carbon (AOC) concentration (17 ± 2 μg C L(-1)) determined by growth of pure cultures in the water after granular activated-carbon filtration (GACF). Growth of the indigenous bacteria measured as adenosine tri-phosphate in water samples incubated at 25 °C confirmed the low AOC in the GACF but revealed the presence of compounds promoting growth after more than one week of incubation. Furthermore, the concentration of particulate organic carbon in the GACF (83 ± 42 μg C L(-1), including 65% carbohydrates) exceeded the AOC concentration. The increased biomass accumulation rate in the continuous biofouling monitor (CBM) at the distribution system reservoir demonstrated the presence of easily biodegradable by-products related to ClO2 dosage to the GACF and in the CBM at 42 km from the treatment plant an iron-associated biomass accumulation was observed. The various methods applied thus distinguished between easily assimilable compounds, biopolymers, slowly biodegradable compounds and biomass-accumulation potential, providing an improved assessment of the biostability of the water. Regrowth of aeromonads may be related to biomass-turnover processes in the distribution system, but establishment of quantitative relationships is needed for

  11. A method for the detection of bacteriophages from ocean water

    NASA Astrophysics Data System (ADS)

    Moebus, K.

    1980-03-01

    A method for the isolation of bacteriophages from ocean water is described. It precludes sample storage before starting phage-enrichment cultures and provides for the use of 3 sub-samples enriched with organic nutrients after 1, 2 and 3 days of incubation. The method was used with samples collected from 6 m below the surface at 48 stations between the European continental shelf and the Sargasso Sea. With 213 among 931 bacterial isolates about 250 strains of bacteriophages were detected by two methods of different sensitivity. From 14 samples taken east of the Azores 115 host bacteria have been found versus only 98 from 34 samples collected at westerly stations. The employment of more than one sub-sample per station as well as the use of more sensitive phage-detection procedures was found to be more advantageous the lower the concentration of cultivatable bacteria in a sample.

  12. Label-free offline versus online activity methods for nucleoside diphosphate kinase b using high performance liquid chromatography.

    PubMed

    Lima, Juliana Maria; Salmazo Vieira, Plínio; Cavalcante de Oliveira, Arthur Henrique; Cardoso, Carmen Lúcia

    2016-08-01

    Nucleoside diphosphate kinase from Leishmania spp. (LmNDKb) has recently been described as a potential drug target to treat leishmaniasis disease. Therefore, screening of LmNDKb ligands requires methodologies that mimic the conditions under which LmNDKb acts in biological systems. Here, we compare two label-free methodologies that could help screen LmNDKb ligands and measure NDKb activity: an offline LC-UV assay for soluble LmNDKb and an online two-dimensional LC-UV system based on LmNDKb immobilised on a silica capillary. The target enzyme was immobilised on the silica capillary via Schiff base formation (to give LmNDKb-ICER-Schiff) or affinity attachment (to give LmNDKb-ICER-His). Several aspects of the ICERs resulting from these procedures were compared, namely kinetic parameters, stability, and procedure steps. Both the LmNDKb immobilisation routes minimised the conformational changes and preserved the substrate binding sites. However, considering the number of steps involved in the immobilisation procedure, the cost of reagents, and the stability of the immobilised enzyme, immobilisation via Schiff base formation proved to be the optimal procedure.

  13. Method and apparatus for hydrogen production from water

    NASA Technical Reports Server (NTRS)

    Muradov, Nazim Z. (Inventor)

    2012-01-01

    A method, apparatuses and chemical compositions are provided for producing high purity hydrogen from water. Metals or alloys capable of reacting with water and producing hydrogen in aqueous solutions at ambient conditions are reacted with one or more inorganic hydrides capable of releasing hydrogen in aqueous solutions at ambient conditions, one or more transition metal compounds are used to catalyze the reaction and, optionally, one or more alkali metal-based compounds. The metal or alloy is preferably aluminum. The inorganic hydride is from a family of complex inorganic hydrides; most preferably, NaBH.sub.4. The transition metal catalyst is from the groups VIII and IB; preferably, Cu and Fe. The alkali metal-based compounds are preferably NaOH, KOH, and the like. Hydrogen generated has a purity of at least 99.99 vol. % (dry basis), and is used without further purification in all types of fuel cells, including the polymer electrolyte membrane (PEM) fuel cell.

  14. Method of burning lightly loaded coal-water slurries

    DOEpatents

    Krishna, C.R.

    1984-07-27

    In a preferred arrangement of the method of the invention, a lightly loaded coal-water slurry, containing in the range of approximately 40% to 52% + 2% by weight coal, is atomized to strip water from coal particles in the mixture. Primary combustor air is forced around the atomized spray in a combustion chamber of a combustor to swirl the air in a helical path through the combustion chamber. A flame is established within the combustion chamber to ignite the stripped coal particles, and flame temperature regulating means are provided for maintaining the flame temperature within a desired predetermined range of temperatures that is effective to produce dry, essentially slag-free ash from the combustion process.

  15. Intramembrane Water Associated with TOAC Spin-Labeled Alamethicin: Electron Spin-Echo Envelope Modulation by D2O

    PubMed Central

    Bartucci, R.; Guzzi, R.; Sportelli, L.; Marsh, D.

    2009-01-01

    Alamethicin is a 20-residue, hydrophobic, helical peptide, which forms voltage-sensitive ion channels in lipid membranes. The helicogenic, nitroxyl amino acid TOAC was substituted isosterically for Aib at residue positions 1, 8, or 16 in a F50/5 alamethicin analog to enable EPR studies. Electron spin-echo envelope modulation (ESEEM) spectroscopy was used to investigate the water exposure of TOAC-alamethicin introduced into membranes of saturated or unsaturated diacyl phosphatidylcholines that were dispersed in D2O. Echo-detected EPR spectra were used to assess the degree of assembly of the peptide in the membrane, via the instantaneous diffusion from intermolecular spin-spin interactions. The profile of residue exposure to water differs between membranes of saturated and unsaturated lipids. In monounsaturated dioleoyl phosphatidylcholine, D2O-ESEEM intensities decrease from TOAC1 to TOAC8 and TOAC16 but not uniformly. This is consistent with a transmembrane orientation for the protoassembled state, in which TOAC16 is located in the bilayer leaflet opposite to that of TOAC1 and TOAC8. Relative to the monomer in fluid bilayers, assembled alamethicin is disposed asymmetrically about the bilayer midplane. In saturated dimyristoyl phosphatidylcholine, the D2O-ESEEM intensity is greatest for TOAC8, indicating a more superficial location for alamethicin, which correlates with the difference in orientation between gel- and fluid-phase membranes found by conventional EPR of TOAC-alamethicin in aligned phosphatidylcholine bilayers. Increasing alamethicin/lipid ratio in saturated phosphatidylcholine shifts the profile of water exposure toward that with unsaturated lipid, consistent with proposals of a critical concentration for switching between the two different membrane-associated states. PMID:19186137

  16. Microbiological water methods: quality control measures for Federal Clean Water Act and Safe Drinking Water Act regulatory compliance.

    PubMed

    Root, Patsy; Hunt, Margo; Fjeld, Karla; Kundrat, Laurie

    2014-01-01

    Quality assurance (QA) and quality control (QC) data are required in order to have confidence in the results from analytical tests and the equipment used to produce those results. Some AOAC water methods include specific QA/QC procedures, frequencies, and acceptance criteria, but these are considered to be the minimum controls needed to perform a microbiological method successfully. Some regulatory programs, such as those at Code of Federal Regulations (CFR), Title 40, Part 136.7 for chemistry methods, require additional QA/QC measures beyond those listed in the method, which can also apply to microbiological methods. Essential QA/QC measures include sterility checks, reagent specificity and sensitivity checks, assessment of each analyst's capabilities, analysis of blind check samples, and evaluation of the presence of laboratory contamination and instrument calibration and checks. The details of these procedures, their performance frequency, and expected results are set out in this report as they apply to microbiological methods. The specific regulatory requirements of CFR Title 40 Part 136.7 for the Clean Water Act, the laboratory certification requirements of CFR Title 40 Part 141 for the Safe Drinking Water Act, and the International Organization for Standardization 17025 accreditation requirements under The NELAC Institute are also discussed. PMID:24830168

  17. Assessment of didecyldimethylammonium chloride as a ballast water treatment method.

    PubMed

    van Slooten, Cees; Peperzak, Louis; Buma, Anita G J

    2015-01-01

    Ballast water-mediated transfer of aquatic invasive species is considered a major threat to marine biodiversity, marine industry and human health. A ballast water treatment is needed to comply with International Maritime Organization (IMO) ballast water discharge regulations. Didecyldimethylammonium chloride (DDAC) was tested for its applicability as a ballast water treatment method. The treatment of the marine phytoplankton species Tetraselmis suecica, Isochrysis galbana and Chaetoceros calcitrans showed that at 2.5 µL L(-1) DDAC was able to inactivate photosystem II (PSII) efficiency and disintegrate the cells after 5 days of dark incubation. The treatment of natural marine plankton communities with 2.5 µL L(-1) DDAC did not sufficiently decrease zooplankton abundance to comply with the IMO D-2 standard. Bivalve larvae showed the highest resistance to DDAC. PSII efficiency was inactivated within 5 days but phytoplankton cells remained intact. Regrowth occurred within 2 days of incubation in the light. However, untreated phytoplankton exposed to residual DDAC showed delayed cell growth and reduced PSII efficiency, indicating residual DDAC toxicity. Natural marine plankton communities treated with 5 µL L(-1) DDAC showed sufficient disinfection of zooplankton and inactivation of PSII efficiency. Phytoplankton regrowth was not detected after 9 days of light incubation. Bacteria were initially reduced due to the DDAC treatment but regrowth was observed within 5 days of dark incubation. Residual DDAC remained too high after 5 days to be safely discharged. Two neutralization cycles of 50 mg L(-1) bentonite were needed to inactivate residual DDAC upon discharge. The inactivation of residual DDAC may seriously hamper the practical use of DDAC as a ballast water disinfectant.

  18. Field Deployable Method for Arsenic Speciation in Water

    PubMed Central

    Voice, Thomas C.; Flores del Pino, Lisveth V.; Havezov, Ivan; Long, David T.

    2010-01-01

    Contamination of drinking water supplies by arsenic is a world-wide problem. Total arsenic measurements are commonly used to investigate and regulate arsenic in water, but it is well understood that arsenic occurs in several chemical forms, and these exhibit different toxicities. It is problematic to use laboratory-based speciation techniques to assess exposure as it has been suggested that the distribution of species is not stable during transport in some types of samples. A method was developed in this study for the on-site speciation of the most toxic dissolved arsenic species: As (III), As (V), monomethylarsonic acid (MMA) and dimethylarsenic acid (DMA). Development criteria included ease of use under field conditions, applicable at levels of concern for drinking water, and analytical performance. The approach is based on selective retention of arsenic species on specific ion-exchange chromatography cartridges followed by selective elution and quantification using graphite furnace atomic absorption spectroscopy. Water samples can be delivered to a set of three cartridges using either syringes or peristaltic pumps. Species distribution is stable at this point, and the cartridges can be transported to the laboratory for elution and quantitative analysis. A set of ten replicate spiked samples of each compound, having concentrations between 1 and 60 µg/L, were analyzed. Arsenic recoveries ranged from 78–112 % and relative standard deviations were generally below 10%. Resolution between species was shown to be outstanding, with the only limitation being that the capacity for As (V) was limited to approximately 50 µg/L. This could be easily remedied by changes in either cartridge design, or the extraction procedure. Recoveries were similar for two spiked hard groundwater samples indicating that dissolved minerals are not likely to be problematic. These results suggest that this methodology can be use for analysis of the four primary arsenic species of concern in

  19. Field Deployable Method for Arsenic Speciation in Water.

    PubMed

    Voice, Thomas C; Flores Del Pino, Lisveth V; Havezov, Ivan; Long, David T

    2011-01-01

    Contamination of drinking water supplies by arsenic is a world-wide problem. Total arsenic measurements are commonly used to investigate and regulate arsenic in water, but it is well understood that arsenic occurs in several chemical forms, and these exhibit different toxicities. It is problematic to use laboratory-based speciation techniques to assess exposure as it has been suggested that the distribution of species is not stable during transport in some types of samples. A method was developed in this study for the on-site speciation of the most toxic dissolved arsenic species: As (III), As (V), monomethylarsonic acid (MMA) and dimethylarsenic acid (DMA). Development criteria included ease of use under field conditions, applicable at levels of concern for drinking water, and analytical performance.The approach is based on selective retention of arsenic species on specific ion-exchange chromatography cartridges followed by selective elution and quantification using graphite furnace atomic absorption spectroscopy. Water samples can be delivered to a set of three cartridges using either syringes or peristaltic pumps. Species distribution is stable at this point, and the cartridges can be transported to the laboratory for elution and quantitative analysis. A set of ten replicate spiked samples of each compound, having concentrations between 1 and 60 µg/L, were analyzed. Arsenic recoveries ranged from 78-112 % and relative standard deviations were generally below 10%. Resolution between species was shown to be outstanding, with the only limitation being that the capacity for As (V) was limited to approximately 50 µg/L. This could be easily remedied by changes in either cartridge design, or the extraction procedure. Recoveries were similar for two spiked hard groundwater samples indicating that dissolved minerals are not likely to be problematic. These results suggest that this methodology can be use for analysis of the four primary arsenic species of concern in

  20. Field deployable method for arsenic speciation in water

    NASA Astrophysics Data System (ADS)

    Voice, Thomas C.; Flores del Pino, Lisveth V.; Havezov, Ivan; Long, David T.

    Contamination of drinking water supplies by arsenic is a world-wide problem. Total arsenic measurements are commonly used to investigate and regulate arsenic in water, but it is well understood that arsenic occurs in several chemical forms, and these exhibit different toxicities. It is problematic to use laboratory-based speciation techniques to assess exposure as it has been suggested that the distribution of species is not stable during transport in some types of samples. A method was developed in this study for the on-site speciation of the most toxic dissolved arsenic species: As (III), As (V), monomethylarsonic acid (MMA) and dimethylarsenic acid (DMA). Development criteria included ease of use under field conditions, applicable at levels of concern for drinking water, and analytical performance. The approach is based on selective retention of arsenic species on specific ion-exchange chromatography cartridges followed by selective elution and quantification using graphite furnace atomic absorption spectroscopy. Water samples can be delivered to a set of three cartridges using either syringes or peristaltic pumps. Species distribution is stable at this point, and the cartridges can be transported to the laboratory for elution and quantitative analysis. A set of ten replicate spiked samples of each compound, having concentrations between 1 and 60 μg/L, were analyzed. Arsenic recoveries ranged from 78% to 112% and relative standard deviations were generally below 10%. Resolution between species was shown to be outstanding, with the only limitation being that the capacity for As (V) was limited to approximately 50 μg/L. This could be easily remedied by changes in either cartridge design, or the extraction procedure. Recoveries were similar for two spiked hard groundwater samples indicating that dissolved minerals are not likely to be problematic. These results suggest that this methodology can be use for analysis of the four primary arsenic species of concern in

  1. Label-free and real-time detection of antigen-antibody interactions by Oblique-incidence Reflectivity Difference (OIRD) method

    NASA Astrophysics Data System (ADS)

    He, LiPing; Sun, Yue; Dai, Jun; Wang, JingYi; Lü, HuiBin; Wang, ShuFang; Jin, KuiJuan; Zhou, YueLiang; Yang, GuoZhen

    2012-09-01

    We label-free and real-time detected three interaction processes of antigen-antibodies, Human Immunoglobulin G (IgG), Rabbit IgG, and Mouse IgG as the targets, and Goat Anti-human IgG, Goat Anti-rabbit IgG, and Goat Anti-mouse IgG as the probe, by the Oblique-incidence Reflectivity Difference (OIRD) method. The interaction dynamic curves of the OIRD signal, corresponding to the interaction processes of antigen-antibodies, are generated. The reaction times from beginning to equilibrium state are about 1800, 900, and 1200 s for Human IgG, Rabbit IgG, and Mouse IgG, respectively. The experimental results demonstrate that the OIRD method not only can distinguish biomolecular interactions, but also can be used in real-time detection of interactions and dynamic processes of biomolecules.

  2. Method and apparatus for acoustic imaging of objects in water

    DOEpatents

    Deason, Vance A.; Telschow, Kenneth L.

    2005-01-25

    A method, system and underwater camera for acoustic imaging of objects in water or other liquids includes an acoustic source for generating an acoustic wavefront for reflecting from a target object as a reflected wavefront. The reflected acoustic wavefront deforms a screen on an acoustic side and correspondingly deforms the opposing optical side of the screen. An optical processing system is optically coupled to the optical side of the screen and converts the deformations on the optical side of the screen into an optical intensity image of the target object.

  3. A method for in situ monitoring of the isotope composition of tree xylem water using laser spectroscopy.

    PubMed

    Volkmann, Till H M; Kühnhammer, Kathrin; Herbstritt, Barbara; Gessler, Arthur; Weiler, Markus

    2016-09-01

    Field studies analyzing the stable isotope composition of xylem water are providing important information on ecosystem water relations. However, the capacity of stable isotopes to characterize the functioning of plants in their environment has not been fully explored because of methodological constraints on the extent and resolution at which samples could be collected and analysed. Here, we introduce an in situ method offering the potential to continuously monitor the stable isotope composition of tree xylem water via its vapour phase using a commercial laser-based isotope analyser and compact microporous probes installed into the xylem. Our technique enables efficient high-frequency measurement with intervals of only a few minutes per sample while eliminating the need for costly and cumbersome destructive collection of plant material and laboratory-based processing. We present field observations of xylem water hydrogen and oxygen isotope compositions obtained over several days including a labelled irrigation event and compare them against results from concurrent destructive sampling with cryogenic distillation and mass spectrometric analysis. The data demonstrate that temporal changes as well as spatial patterns of integration in xylem water isotope composition can be resolved through direct measurement. The new technique can therefore present a valuable tool to study the hydraulic architecture and water utilization of trees.

  4. A method for in situ monitoring of the isotope composition of tree xylem water using laser spectroscopy.

    PubMed

    Volkmann, Till H M; Kühnhammer, Kathrin; Herbstritt, Barbara; Gessler, Arthur; Weiler, Markus

    2016-09-01

    Field studies analyzing the stable isotope composition of xylem water are providing important information on ecosystem water relations. However, the capacity of stable isotopes to characterize the functioning of plants in their environment has not been fully explored because of methodological constraints on the extent and resolution at which samples could be collected and analysed. Here, we introduce an in situ method offering the potential to continuously monitor the stable isotope composition of tree xylem water via its vapour phase using a commercial laser-based isotope analyser and compact microporous probes installed into the xylem. Our technique enables efficient high-frequency measurement with intervals of only a few minutes per sample while eliminating the need for costly and cumbersome destructive collection of plant material and laboratory-based processing. We present field observations of xylem water hydrogen and oxygen isotope compositions obtained over several days including a labelled irrigation event and compare them against results from concurrent destructive sampling with cryogenic distillation and mass spectrometric analysis. The data demonstrate that temporal changes as well as spatial patterns of integration in xylem water isotope composition can be resolved through direct measurement. The new technique can therefore present a valuable tool to study the hydraulic architecture and water utilization of trees. PMID:27260852

  5. Determination of microcystin-LR in water by a label-free aptamer based electrochemical impedance biosensor.

    PubMed

    Lin, Zhenyu; Huang, Huiming; Xu, Yixiang; Gao, Xiaoyao; Qiu, Bin; Chen, Xi; Chen, Guonan

    2013-01-15

    In this study, an electrochemical impedance biosensor for cyanobacterial toxin microcystin-LR (MC-LR) detection has been developed. MC-LR aptamers were immobilized on the gold electrode through Au-S interaction, in the presence of target (MC-LR); the binding of MC-LR and aptamers probe led to a complex formation change on the electrode surface and resulted in the impedance decreasing. The decrease rate had a linear relationship with logarithm of the MC-LR concentration in the range of 1.0 × 10(-7)-5.0 × 10(-11)mol/L, with a detection limit of 1.8 × 10(-11)mol/L. The sensor has good selectivity and stability, it has been applied to detect MC-LR in three kinds of real water samples with satisfying results.

  6. Instream water use in the United States: water laws and methods for determining flow requirements

    USGS Publications Warehouse

    Lamb, Berton L.; Doerksen, Harvey R.

    1987-01-01

    the conterminous United States consists of about 12,000 miles of maintained waterways, over which about 500 million tons of cargo is carried each year (U.S. Army Corps of Engineers, 1988, p. 16). Although not so widely practiced in recent years, streams have been used to dispose of raw waste products from homes, communities, and factories. This use has been discouraged by law and public policy because of public health concerns and the damage it causes to the environment. Beginning in the mid-1960's, other instream uses gained new prominence in the water-resources arena-the assertion of a legal right to a free-flowing stream for biological, recreational, and esthetic purposes. These uses themselves, however, are not new. Riverine habitat always has produced fish, and the beauty of flowing water always has evoked a strong sense of esthetic appreciation. What is new is the emerging legitimacy and awareness of these noneconomic uses under State and Federal laws and regulations. In the past, environmental uses of flowing water were ignored, for the most part, under a long-standing legal tradition that favored offstream uses and certain instream uses that had a strong economic basis. The history of instream-flow policy debate really concerns those recently recognized types of interim uses. Although the more transitional water uses have been protected by law, the recognition of other in stream uses has resulted in substantial changes in State water laws. Although methods for determining the volume of water needed for most traditional water uses are relatively straight-forward and well-established, methods for determining water requirements for the in stream uses have been developed only recently and are continuing to evolve. Water laws that have favored the more traditional water uses, the inherent nature of conflict between instream and offstream water uses, and the special kinds of technological and philosophical problems posed by the "newer" types of instream uses are

  7. Direct method for detection and characterization of cell surface receptors for insulin by means of 125I-labeled autoantibodies against the insulin receptor.

    PubMed Central

    Jarrett, D B; Roth, J; Kahn, C R; Flier, J S

    1976-01-01

    Autoantibodies directed against the cell surface receptors for insulin are found in some patients with extreme insulin resistance. These antibodies specifically inhibit the binding of insulin to its receptor. A purified IgG fraction from one patient's plasma was labeled with 125I. The 125I-labeled antireceptor antibody, which initially represented about 0.3% of the total 125I-IgG, was enriched by selective adsorption and subsequent elution from cells rich in insulin receptors. The 125I-antireceptor antibody bound to cells and the binding was inhibited by whole plasma and purified IgG from this patient, as well as whole plasma from another patient with autoantibodies to the insulin receptor. Insulins that differed 300-fold in biological potency and affinity inhibited binding of 125I-antireceptor antibody in direct proportion to their ability to bind to the insulin receptor. The binding of 125I-antireceptor antibody was closely correlated with the binding of 125I-insulin over a wide range of receptor concentrations on different cell types. Experimentally induced reduction of the insulin receptor concentration was associated with parallel decreases in the binding of 125I-antireceptor antibody and 125I-insulin. The preparation of 125I-antireceptor antibody with a high specific activity by cytoadsorption and elution has provided a sensitive method for the detection of receptors and autoantibodies to cell surface components. PMID:1069300

  8. Investigating an organ-targeting platform based on hydroxyapatite nanoparticles using a novel in situ method of radioactive ¹²⁵Iodine labeling.

    PubMed

    Ignjatović, Nenad; Vranješ Djurić, Sanja; Mitić, Zarko; Janković, Drina; Uskoković, Dragan

    2014-10-01

    In this study, we have investigated the synthesis of nanoparticles of hydroxyapatite (HAp) and hydroxyapatite coated with chitosan (HAp/Ch) and the chitosan-poly-d,l-lactide-co-glycolide polymer blend (HAp/Ch-PLGA) as an organ-targeting system. We have examined and defined the final destination, as well as the dynamics and the pathways of the synthesized particles following intravenous administration in vivo. The XRD, ZP, FT-IR and SEM analyses have confirmed that the hydroxyapatite nanoparticles with d50=72 nm are coated with polymers. Radioactive 125-Iodine ((125)I), a low energy gamma emitter, was used to develop a novel in situ method for the radiolabeling of particles and investigation of their biodistribution. (125)I-labeled particles exhibited high stability in saline and serum over the second day, which justified their use in the following in vivo studies. The biodistribution of (125)I-labeled particles after intravenous injection in rats differed significantly: HAp particles mostly targeted the liver, HAp/Ch the spleen and the liver, while HAp/Ch-PLGA targeted the lungs. Twenty-four hours post injection, HAp particles were excreted completely, while both (125)I-HAp/Ch and (125)I-HAp/Ch-PLGA were retained in the body for a prolonged period of time with more than 20% of radioactivity still found in different organs. PMID:25175234

  9. High performance concentration method for viruses in drinking water.

    PubMed

    Kunze, Andreas; Pei, Lu; Elsässer, Dennis; Niessner, Reinhard; Seidel, Michael

    2015-09-15

    According to the risk assessment of the WHO, highly infectious pathogenic viruses like rotaviruses should not be present in large-volume drinking water samples of up to 90 m(3). On the other hand, quantification methods for viruses are only operable in small volumes, and presently no concentration procedure for processing such large volumes has been reported. Therefore, the aim of this study was to demonstrate a procedure for processing viruses in-line of a drinking water pipeline by ultrafiltration (UF) and consecutive further concentration by monolithic filtration (MF) and centrifugal ultrafiltration (CeUF) of viruses to a final 1-mL sample. For testing this concept, the model virus bacteriophage MS2 was spiked continuously in UF instrumentation. Tap water was processed in volumes between 32.4 m(3) (22 h) and 97.7 m(3) (72 h) continuously either in dead-end (DE) or cross-flow (CF) mode. Best results were found by DE-UF over 22 h. The concentration of MS2 was increased from 4.2×10(4) GU/mL (genomic units per milliliter) to 3.2×10(10) GU/mL and from 71 PFU/mL to 2×10(8) PFU/mL as determined by qRT-PCR and plaque assay, respectively.

  10. Differential mass spectrometry: a label-free LC-MS method for finding significant differences in complex peptide and protein mixtures.

    PubMed

    Wiener, Matthew C; Sachs, Jeffrey R; Deyanova, Ekaterina G; Yates, Nathan A

    2004-10-15

    Efficiently identifying and quantifying disease- or treatment-related changes in the abundance of proteins is an important area of research for the pharmaceutical industry. Here we describe an automated, label-free method for finding differences in complex mixtures using complete LC-MS data sets, rather than subsets of extracted peaks or features. The method selectively finds statistically significant differences in the intensity of both high-abundance and low-abundance ions, accounting for the variability of measured intensities and the fact that true differences will persist in time. The method was used to compare two complex peptide mixtures with known peptide differences. This controlled experiment allowed us to assess the validity of each difference found and so to analyze the method's sensitivity and specificity. The method detects both presence versus absence and a 2-fold change in peptide concentration near the limit of detection of the instrument used, where chromatographic peaks may not be sufficiently well defined to be detected in individual samples. The method is more sensitive and gives fewer false positives than subtractive methods that ignore signal variability. Differential mass spectrometry combined with targeted MS/MS analysis of only identified differences may save both computation time and human effort compared to shotgun proteomics approaches. PMID:15481957

  11. 33 CFR 151.2035 - Implementation schedule for approved ballast water management methods.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... SUBSTANCES, GARBAGE, MUNICIPAL OR COMMERCIAL WASTE, AND BALLAST WATER Ballast Water Management for Control of Nonindigenous Species in Waters of the United States § 151.2035 Implementation schedule for approved ballast water management methods. (a) To discharge ballast water into waters of the United States, the...

  12. RAPID SEPARATION METHOD FOR EMERGENCY WATER AND URINE SAMPLES

    SciTech Connect

    Maxwell, S.; Culligan, B.

    2008-08-27

    The Savannah River Site Environmental Bioassay Lab participated in the 2008 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2008. A new rapid column separation method was used for analysis of actinides and {sup 90}Sr the NRIP 2008 emergency water and urine samples. Significant method improvements were applied to reduce analytical times. As a result, much faster analysis times were achieved, less than 3 hours for determination of {sup 90}Sr and 3-4 hours for actinides. This represents a 25%-33% improvement in analysis times from NRIP 2007 and a {approx}100% improvement compared to NRIP 2006 report times. Column flow rates were increased by a factor of two, with no significant adverse impact on the method performance. Larger sample aliquots, shorter count times, faster cerium fluoride microprecipitation and streamlined calcium phosphate precipitation were also employed. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and {sup 90}Sr analyses for NRIP 2008 emergency urine samples. High levels of potential matrix interferences may be present in emergency samples and rugged methods are essential. Extremely high levels of {sup 210}Po were found to have an adverse effect on the uranium results for the NRIP-08 urine samples, while uranium results for NRIP-08 water samples were not affected. This problem, which was not observed for NRIP-06 or NRIP-07 urine samples, was resolved by using an enhanced {sup 210}Po removal step, which will be described.

  13. Method of generating hydrogen by catalytic decomposition of water

    DOEpatents

    Balachandran, Uthamalingam; Dorris, Stephen E.; Bose, Arun C.; Stiegel, Gary J.; Lee, Tae-Hyun

    2002-01-01

    A method for producing hydrogen includes providing a feed stream comprising water; contacting at least one proton conducting membrane adapted to interact with the feed stream; splitting the water into hydrogen and oxygen at a predetermined temperature; and separating the hydrogen from the oxygen. Preferably the proton conducting membrane comprises a proton conductor and a second phase material. Preferable proton conductors suitable for use in a proton conducting membrane include a lanthanide element, a Group VIA element and a Group IA or Group IIA element such as barium, strontium, or combinations of these elements. More preferred proton conductors include yttrium. Preferable second phase materials include platinum, palladium, nickel, cobalt, chromium, manganese, vanadium, silver, gold, copper, rhodium, ruthenium, niobium, zirconium, tantalum, and combinations of these. More preferably second phase materials suitable for use in a proton conducting membrane include nickel, palladium, and combinations of these. The method for generating hydrogen is preferably preformed in the range between about 600.degree. C. and 1,700.degree. C.

  14. A rapid improved method for sexing embryo of water buffalo.

    PubMed

    Zoheir, K M A; Allam, A A

    2011-07-01

    The objective of the experiment of this paper is to develop and improve in the sexing method for preimplantation embryos of water buffalo (Bubalus bubalis) using loop-mediated isothermal amplification (LAMP) reaction. Embryo sexing has been recognized to control effectively the sex of offspring in the embryo transfer industry. A rapid and simple detection system was established by adding ethidium bromide (EB) or 5 μl of CuSO4 (3M) to the product of LAMP reaction. The result of these additions after 2 min was a color change and a precipitate. It could be employed as an alternative method in the detection of the reaction products in place of the time consuming electrophoresis or the turbidity meter. The in vitro produced buffalo embryos were divided into one to eight pieces using a microblade attached to a micromanipulator. The cell number in each piece was counted before sexing. Sexing of DNA samples extracted from one to five biopsies cells was performed by LAMP. After biopsy, the remaining part of the embryos was used to confirm the sex by polymerase chain reaction (PCR). Fifty buffalo embryos were used and the accuracy of sex prediction was 100% when the blastomeres dissociated from a morula exceeds three. In conclusion, the present procedure without turbidity meter and electrophoresis was reliable and applicable for sexing the water buffalo embryos.

  15. Evaluation of Water Treatment Methods for Endocrine Disrupting Compounds

    NASA Astrophysics Data System (ADS)

    Thomas, S. M.; Murray, K. E.

    2006-05-01

    Endocrine disrupting compounds (EDCs) have caught recent attention as one of the major concerns in the environment. They are known to interfere with the activity of growth-related hormones and usually, as a result, cause disruption in normal functioning of the body. The compounds currently classified as EDCs range from a variety of both natural and synthetic organic compounds and also some heavy metals. Most of these compounds are used in household, pharmaceutical, industrial, agricultural activities, the consumption or usage of which increases with population. There is a lack of detailed chemical and biological analysis as to what concentrations each of these EDCs pose harmless to the environment because of the large number of the suspected compounds. However, several published reports have established that endocrine disruption is observed in aquatic species due to chronic exposure to concentrations of some EDCs as low as a few ng/l. Conventional water treatment facilities do not usually suffice to remove EDCs in concentrations below 1 ng/l. Available technologies for removal of EDCs include adsorption, degradation and membrane treatment. The removal rates, however, are dependant on the properties of the compound, such as molecular weight, water- octanol partition coefficient and vapor pressure; physiochemical conditions of the matrix such as, redox and temperature conditions; type and dose of degrading agent and the concentration of the EDCs. Since, EDCs comprise a vast variety of compounds, their response to each of these treatment methods will be different and hence it is plausible that a single treatment technique will not be sufficient to remove the EDCs to very low concentrations. Based on our review of existing water treatment methods, we believe that a sequential treatment technique that consists of an adsorption, a degradation and finally a fine membrane treatment, each optimized for favorable, efficient and inexpensive removal may be required to remove

  16. Final Report for research grant "Development of Methods for High Specific Activity Labeling of Biomolecules Using Astatine-211 in Different Oxidation States"

    SciTech Connect

    Wilbur, D., Scott

    2011-12-14

    The overall objective of this research effort was to develop methods for labeling biomolecules with higher oxidation state species of At-211. This was to be done in an effort to develop reagents that had higher in vivo stability than the present carbon-bonded At-211-labeled compounds. We were unsuccessful in that effort, as none of the approaches studied provided reagents that were stable to in vivo deastatination. However, we gained a lot of information about At-211 in higher oxidation states. The studies proved to be very difficult as small changes in pH and other conditions appeared to change the nature of the species that obtained (by HPLC retention time analyses), with many of the species being unidentifiable. The fact that there are no stable isotopes of astatine, and the chemistry of the nearest halogen iodine is quite different, made it very difficult to interpret results of some experiments. With that said, we believe that a lot of valuable information was obtained from the studies. The research effort evaluated: (1) methods for chemical oxidation of At-211, (2) approaches to chelation of oxidized At-211, and (3) approaches to oxidation of astatophenyl compounds. A major hurdle that had to be surmounted to conduct the research was the development of HPLC conditions to separate and identify the various oxidized species formed. Attempts to develop conditions for separation of iodine and astatine species by normal and reversed-phase TLC and ITLC were not successful. However, we were successful in developing conditions (from a large number of attempts) to separate oxidized forms of iodine ([I-125]iodide, [I-125]iodate and [I-125]periodate) and astatine ([At-211]astatide, [At-211]astatate, [At-211]perastatate, and several unidentified At-211 species). Information on the basic oxidation and characterization of At-211 species is provided under Objective 1. Conditions were developed to obtain new At-211 labeling method where At-211 is chelated with the DOTA and

  17. Improved HPLC Method Using 2,3-naphthalenedicarboxaldehyde as Fluorescent Labeling Agent for Quantification of Histamine in Human Immunoglobulin Preparations

    PubMed Central

    Kim, Jung-Hwan; Shin, In Soo; Lee, Yoo Kyoung; Oh, Ho Jung; Ban, Sang Ja

    2011-01-01

    Objectives To develop and optimize quantitative HPLC method using 2,3-naphthalenedicarboxaldehyde (NDA) after simple and efficient solid phase extraction to determine the histamine in a biopharmaceutical (Histobulin™). Methods The HPLC method was established using NDA-induced Histobulin and compared with the recently reported HPLC method using o-phthaldehyde (OPA). The validated NDA-applied HPLC method was adjusted to 15 lots of Histobulin and compared by the current lot-release-test method using fluorimetry in recovery of histamine and reproducibility. Results Analyses of six HPLC chromatograms using NDA and OPA each were compared. NDA produced a more stable chromatogram baseline than OPA, and showed better stability. The HPLC analysis was validated in accuracy (91–103%), precision (interday/intraday assay CV ≤2.30%), and linearity of dose–response curve (R2 ≥ 0.9919). The detection limit was 0.0076 μg/mL and the quantitative limit was 0.0229 μg/mL. The amount of histamine per 12 mg of immunoglobulin was determined to be 0.17 ± 0.016 μg by the HPLC and 0.025 ± 0.013 μg by the current lot-release-test method using fluorimetry. Conclusion NDA derivatization showed better stability compared with the OPA method. Therefore the newly established NDA-derivatizated HPLC method may be more suitable than the fluorimetric method in lot-release-tests of biopharmaceuticals. PMID:24159462

  18. 78 FR 22540 - Notice of Public Meeting/Webinar: EPA Method Development Update on Drinking Water Testing Methods...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-16

    ... AGENCY Notice of Public Meeting/Webinar: EPA Method Development Update on Drinking Water Testing Methods...: Notice of public meeting. SUMMARY: The U.S. Environmental Protection Agency (EPA) Office of Ground Water and Drinking Water, Standards and Risk Management Division's Technical Support Center (TSC)...

  19. Drinking water test methods in crisis-afflicted areas: comparison of methods under field conditions.

    PubMed

    Merle, Roswitha; Bleul, Ingo; Schulenburg, Jörg; Kreienbrock, Lothar; Klein, Günter

    2011-11-01

    To simplify the testing of drinking water in crisis-afflicted areas (as in Kosovo in 2007), rapid test methods were compared with the standard test. For Escherichia coli and coliform pathogens, rapid tests were made available: Colilert(®)-18, P/A test with 4-methylumbelliferyl-β-D-glucoronid, and m-Endo Broth. Biochemical differentiation was carried out by Enterotube™ II. Enterococci were determined following the standard ISO test and by means of Enterolert™. Four hundred ninety-nine water samples were tested for E. coli and coliforms using four methods. Following the standard method, 20.8% (n=104) of the samples contained E. coli, whereas the rapid tests detected between 19.6% (m-Endo Broth, 92.0% concordance) and 20.0% (concordance: 93.6% Colilert-18 and 94.8% P/A-test) positive samples. Regarding coliforms, the percentage of concordant results ranged from 98.4% (P/A-test) to 99.0% (Colilert-18). Colilert-18 and m-Endo Broth detected even more positive samples than the standard method did. Enterococci were detected in 93 of 573 samples by the standard method, but in 92 samples by Enterolert (concordance: 99.5%). Considering the high-quality equipment and time requirements of the standard method, the use of rapid tests in crisis-afflicted areas is sufficiently reliable.

  20. Fully automated TV-image analysis of the cell-cycle: comparison of the PLM method with determinations of the percentage and the DNA content of labelled cells.

    PubMed

    Wachsmuth, E D; van Golitschek, M; Macht, F; Maurer-Schultze, B

    1988-01-01

    A cell-cycle analysis based on a fully automated TV-image scanning system is proposed to replace the laborious PLM method. To compare the efficiency of the two procedures, cell-cycle parameters were assessed in Ehrlich (diploid and hyperdiploid), L-1210, and JB-1 mouse ascites tumours and in rat jejunal crypts. The percentages of labelled mitoses (PLM) were counted visually on Feulgen-stained autoradiographs obtained at various times after a single 3H-thymidine pulse. The fraction of labelled cells (P) and the DNA ratio of labelled and unlabelled cells were measured by TV-image analysis in the same slides and plotted against time. Within practical limits, TV-image analysis using the P-curve gives the same results as the PLM method. Using the P-curve has the important advantage that its first part, beginning at the time of 3H-thymidine injection and ending at the first maximum, furnishes more information about the cell cycle than the corresponding part of the PLM curve. It can be used to compute tG2M tS and the ratio of the growth faction index to the cell-cycle time (IP/tC) whereas the first part of the PLM-curve reveals only the length of the S-phase (tS). The IP/tC ratio is a readily accessible measure of growth and increases when the cells divide more frequently. Cell death rates may be neglected since the ratio is determined within less than the duration of one cell cycle. Moreover, the data from the first part of the P curve indicate whether there is a large non-growth fraction. If the non-growth fraction is small, i.e. if IP approximately 1, the P curve need only be measured until the first maximum is reached so that fewer samples and animals are required. If the non-growth fraction is large or unknown, the cell-cycle parameters are calculated by reference to the position and size not only of the first minimum and the first maximum, but also of the second minimum of the P curve.