Deep freezing of Angora goat semen: effects of diluent composition and method and rate of dilution on survival of spermatozoa.

PubMed

Salamon, S; Ritar, A J

1982-01-01

Five factorial experiments were conducted to examine the effects of concentration of tris(hydroxymethyl)aminomethane (Tris), type and concentration of sugar in the diluent, rate and method of dilution on the survival of goat spermatozoa after freezing by the pellet method. Spermatozoa tolerated a relatively wide range in concentration of Tris, but the cell survival depended on the type of sugar included in the Tris diluent. Glucose and fructose were more suitable components than lactose or raffinose. Survival of spermatozoa after thawing was better for three- to fivefold prefreezing dilution. There was interaction between method of semen dilution (one-step, two-step), holding time at 5 degrees C, and glycerol concentration. The best result was obtained after one-step dilution at 30 degrees C (Tris 375 mM-glucose 41.625 mM-citric acid 124 mM), 1.5 h holding at 5 degrees C, and with 4% (v/v) glycerol concentration in the diluted semen.

Estimation method for serial dilution experiments.

PubMed

Ben-David, Avishai; Davidson, Charles E

2014-12-01

Titration of microorganisms in infectious or environmental samples is a corner stone of quantitative microbiology. A simple method is presented to estimate the microbial counts obtained with the serial dilution technique for microorganisms that can grow on bacteriological media and develop into a colony. The number (concentration) of viable microbial organisms is estimated from a single dilution plate (assay) without a need for replicate plates. Our method selects the best agar plate with which to estimate the microbial counts, and takes into account the colony size and plate area that both contribute to the likelihood of miscounting the number of colonies on a plate. The estimate of the optimal count given by our method can be used to narrow the search for the best (optimal) dilution plate and saves time. The required inputs are the plate size, the microbial colony size, and the serial dilution factors. The proposed approach shows relative accuracy well within ±0.1log10 from data produced by computer simulations. The method maintains this accuracy even in the presence of dilution errors of up to 10% (for both the aliquot and diluent volumes), microbial counts between 10(4) and 10(12) colony-forming units, dilution ratios from 2 to 100, and plate size to colony size ratios between 6.25 to 200. Published by Elsevier B.V.

Videodensitometric Methods for Cardiac Output Measurements

NASA Astrophysics Data System (ADS)

Mischi, Massimo; Kalker, Ton; Korsten, Erik

2003-12-01

Cardiac output is often measured by indicator dilution techniques, usually based on dye or cold saline injections. Developments of more stable ultrasound contrast agents (UCA) are leading to new noninvasive indicator dilution methods. However, several problems concerning the interpretation of dilution curves as detected by ultrasound transducers have arisen. This paper presents a method for blood flow measurements based on UCA dilution. Dilution curves are determined by real-time densitometric analysis of the video output of an ultrasound scanner and are automatically fitted by the Local Density Random Walk model. A new fitting algorithm based on multiple linear regression is developed. Calibration, that is, the relation between videodensity and UCA concentration, is modelled by in vitro experimentation. The flow measurement system is validated by in vitro perfusion of SonoVue contrast agent. The results show an accurate dilution curve fit and flow estimation with determination coefficient larger than 0.95 and 0.99, respectively.

Semiquantitative determination of mesophilic, aerobic microorganisms in cocoa products using the Soleris NF-TVC method.

PubMed

Montei, Carolyn; McDougal, Susan; Mozola, Mark; Rice, Jennifer

2014-01-01

The Soleris Non-fermenting Total Viable Count method was previously validated for a wide variety of food products, including cocoa powder. A matrix extension study was conducted to validate the method for use with cocoa butter and cocoa liquor. Test samples included naturally contaminated cocoa liquor and cocoa butter inoculated with natural microbial flora derived from cocoa liquor. A probability of detection statistical model was used to compare Soleris results at multiple test thresholds (dilutions) with aerobic plate counts determined using the AOAC Official Method 966.23 dilution plating method. Results of the two methods were not statistically different at any dilution level in any of the three trials conducted. The Soleris method offers the advantage of results within 24 h, compared to the 48 h required by standard dilution plating methods.

Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples.

PubMed

Thomas, Pious; Sekhar, Aparna C; Upreti, Reshmi; Mujawar, Mohammad M; Pasha, Sadiq S

2015-12-01

We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single plate-serial dilution spotting (SP-SDS) with the prime recommendation of sample anchoring (10 0 stocks). For pure cultures, serial dilutions were prepared from 0.1 OD (10 0 ) stock and 20 μl aliquots of six dilutions (10 1 -10 6 ) were applied as 10-15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 10 0 -10 5 dilutions, and for colloidal suspensions and solid samples (10% w/v), 10 1 -10 6 dilutions were used. Following incubation, at least one dilution level yielded 6-60 cfu per sector comparable to the standard method involving 100 μl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae , SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.

Body composition among Sri Lankan infants by 18*O dilution method and the validity of anthropometric equations to predict body fat against 18*O dilution

USDA-ARS?s Scientific Manuscript database

Body composition indicators provide a better guidance for growth and nutritional status of the infants. This study was designed to (1) measure the body composition of the Sri Lankan infants using a reference method, the 18*O dilution method; (2) calculate the body fat content of the infants using pu...

Methods for deoxygenating biomass-derived pyrolysis oil

DOEpatents

Brandvold, Timothy A.

2015-07-14

Methods for deoxygenating a biomass-derived pyrolysis oil are provided. A method comprising the steps of diluting the biomass-derived pyrolysis oil with a phenolic-containing diluent to form a diluted pyoil-phenolic feed is provided. The diluted pyoil-phenolic feed is contacted with a deoxygenating catalyst in the presence of hydrogen at hydroprocessing conditions effective to form a low-oxygen biomass-derived pyrolysis oil effluent.

Comparison of gravimetric and a double-indicator dilution technique for assessment of extra-vascular lung water in endotoxaemia.

PubMed

Rossi, P; Oldner, A; Wanecek, M; Leksell, L G; Rudehill, A; Konrad, D; Weitzberg, E

2003-03-01

To compare a molecular double-indicator dilution technique with the gravimetrical reference method for measurement of extra-vascular lung water in porcine endotoxin shock. Open comparative experimental study. Animal research laboratory. In fourteen anaesthetised, mechanically ventilated landrace pigs, central and pulmonary haemodynamics as well as pulmonary gas exchange were measured. Extra-vascular lung water was quantitated gravimetrically as well as with a molecular double indicator dilution technique. Eight of these animals were subjected to endotoxaemia, the rest serving as sham controls. No difference in extra-vascular lung water was observed between the two methods in sham animals. Furthermore, extra-vascular lung water assessed with the molecular double-indicator dilution technique at the initiation of endotoxin infusion did not differ significantly from the corresponding values for sham animals. Endotoxaemia induced a hypodynamic shock with concurrent pulmonary hypertension and a pronounced deterioration in gas exchange. No increase in extra-vascular lung water was detected with the molecular double-indicator dilution technique in response to endotoxin, whereas this parameter was significantly higher when assessed with the gravimetric method. The molecular double-indicator dilution technique showed similar results as the gravimetrical method for assessment of extra-vascular lung water in non-endotoxaemic conditions. However, during endotoxin-induced lung injury the molecular double indicator dilution technique failed to detect the significant increase in extra-vascular lung water as measured by the gravimetric method. These data suggest that the molecular double indicator dilution technique may be of limited value during sepsis-induced lung injury.

Comparison of Anaerobic Susceptibility Results Obtained by Different Methods

PubMed Central

Rosenblatt, J. E.; Murray, P. R.; Sonnenwirth, A. C.; Joyce, J. L.

1979-01-01

Susceptibility tests using 7 antimicrobial agents (carbenicillin, chloramphenicol, clindamycin, penicillin, cephalothin, metronidazole, and tetracycline) were run against 35 anaerobes including Bacteroides fragilis (17), other gram-negative bacilli (7), clostridia (5), peptococci (4), and eubacteria (2). Results in triplicate obtained by the microbroth dilution method and the aerobic modification of the broth disk method were compared with those obtained with an agar dilution method using Wilkins-Chalgren agar. Media used in the microbroth dilution method included Wilkins-Chalgren broth, brain heart infusion broth, brucella broth, tryptic soy broth, thioglycolate broth, and Schaedler's broth. A result differing by more than one dilution from the Wilkins-Chalgren agar result was considered a discrepancy, and when there was a change in susceptibility status this was termed a significant discrepancy. The microbroth dilution method using Wilkins-Chalgren broth and thioglycolate broth produced the fewest total discrepancies (22 and 24, respectively), and Wilkins-Chalgren broth, thioglycolate, and Schaedler's broth had the fewest significant discrepancies (6, 5, and 5, respectively). With the broth disk method, there were 15 significant discrepancies, although half of these were with tetracycline, which was the antimicrobial agent associated with the highest number of significant discrepancies (33), considering all of the test methods and media. PMID:464560

Doing More with Less? Toward Increasing the Resolution of Protistan Grazing-rate Measurements.

NASA Astrophysics Data System (ADS)

Morison, F.; Menden-Deuer, S.

2016-02-01

The dilution method is the standard protocol to quantify phytoplankton grazing-mortality rates and has been key in developing an understanding of protistan grazing impact on ocean primary production. Although the method's extensive use has facilitated the acquisition of a global dataset, its laborious application hinders the sampling resolution needed to fill knowledge gaps remaining at the geographical, seasonal, and vertical scales, and of the effects of climate-related factors influencing grazing magnitude. Here we present a rigorous assessment of an abbreviated method known as the 2-point. We analyzed unpublished results from 77 dilution experiments performed using a series of up to 5 dilutions under a wide range of chlorophyll concentrations and temperatures. We quantified the difference between estimates of both phytoplankton growth and grazing-mortality obtained based on the full dilution series and those obtained when the number of dilutions was reduced to 2. We considered the effect of non-linearity and chlorophyll concentration, and generated quantified estimates of trade-offs when choosing the fraction of seawater in the diluted treatment. Ultimately, we provide an assessment of the reliability of the 2-point method and recommendations on how to apply it.

Methods and apparatuses for deoxygenating biomass-derived pyrolysis oil

DOEpatents

Baird, Lance Awender; Brandvold, Timothy A.

2015-10-20

Embodiments of methods and apparatuses for deoxygenating a biomass-derived pyrolysis oil are provided. In one example, a method comprises the steps of separating a low-oxygen biomass-derived pyrolysis oil effluent into a low-oxygen-pyoil organic phase stream and an aqueous phase stream. Phenolic compounds are removed from the aqueous phase stream to form a phenolic-rich diluent recycle stream. A biomass-derived pyrolysis oil stream is diluted and heated with the phenolic-rich diluent recycle stream to form a heated diluted pyoil feed stream. The heated diluted pyoil feed stream is contacted with a deoxygenating catalyst in the presence of hydrogen to deoxygenate the heated diluted pyoil feed stream.

The development of a practicable method for deepfreezing of boar spermatozoa.

PubMed

Larsson, K; Einarsson, S; Swensson, T

1977-03-01

The present study was made in order to develop the freezing method of Crabo & Einarsson (1971) into a practicable deep-freezing method for boar spermatozoa. Semen was concentrated before dilution and a dosewise thawing procedure for the pellet-frozen semen was utilized. Two fertility tests on altogther 68 gilts were performed. In the first trial the insemination doses contained 6x10(9) spermatozoa and the dilution rate before cooling was 3:2 with a total dilution before freezing of 3:7. In the second trial the insemination doses contained 4.5x10(9) spermatozoa, primary dilution was 3:3 and final dilution 3:7. The pregnancy rate in trial 1 was 53% and in trial 2 72%. Mean litter size in pregnant gilts was 6.5 in trial 1 and 9.7 in trial 2. The difference in ratio of foetuses to c.l. in pregnant gilts between the trials was highly significant. The different dilution procedures are discussed and the lower fertility of semen frozen in trial 1 is suggested to be due to a shortened fertile life of spermatozoa so treated. The freezing method utilized in trial 2 is considered to be practically acceptable and the results are concluded to be satisfactory. A detailed description of the method and the equipment utilized can be requested from the authors.

Production and distribution of dilute species in semiconducting materials

DOEpatents

James, Ralph B.; Camarda, Giuseppe; Bolotnikov, Aleksey E.; Hossain, Anwar; Yang, Ge; Kim, Kihyun

2016-09-06

Technologies are described effective to implement systems and methods of producing a material. The methods comprise receiving a tertiary semiconductor sample with a dilute species. The sample has two ends. The first end of the sample includes a first concentration of the dilute species lower than a second concentration of the dilute species in the second end of the sample. The method further comprises heating the sample in a chamber. The chamber has a first zone and a second zone. The first zone having a first temperature higher than a second temperature in the second zone. The sample is orientated such that the first end is in the first zone and the second end is in the second zone.

Screening fungicides for use in fish culture: Evaluation of the agar plug transfer, cellophane transfer, and agar dilution methods

USGS Publications Warehouse

Bailey, Tom A.

1983-01-01

The reliability, reproducibility, and usefulness of three screening methods -- the cellophane transfer, the agar plug transfer, and the agar dilution -- to screen aquatic fungicides were evaluated. Achlya flagellata and Saprolegnia hypogyna were exposed to 1, 10, and 100 mg/L of malachite green to test each method. The cellophane transfer and agar plug transfer techniques had similar reliability and reproducibility in rating fungicidal activity, and were both superior to the agar dilution technique. The agar plug transfer and agar dilution techniques adequately projected in vivo activity of malachite green, but the cellophane transfer technique overestimated its activity. Overall, the agar plug transfer technique most accurately rated the activity of malachite green and was the easiest test to perform. It therefore appears to be the method of choice for testing aquatic fungicides.

Multi-laboratory survey of qPCR enterococci analysis method performance

EPA Pesticide Factsheets

Quantitative polymerase chain reaction (qPCR) has become a frequently used technique for quantifying enterococci in recreational surface waters, but there are several methodological options. Here we evaluated how three method permutations, type of mastermix, sample extract dilution and use of controls in results calculation, affect method reliability among multiple laboratories with respect to sample interference. Multiple samples from each of 22 sites representing an array of habitat types were analyzed using EPA Method 1611 and 1609 reagents with full strength and five-fold diluted extracts. The presence of interference was assessed three ways: using sample processing and PCR amplifications controls; consistency of results across extract dilutions; and relative recovery of target genes from spiked enterococci in water sample compared to control matrices with acceptable recovery defined as 50 to 200%. Method 1609, which is based on an environmental mastermix, was found to be superior to Method 1611, which is based on a universal mastermix. Method 1611 had over a 40% control assay failure rate with undiluted extracts and a 6% failure rate with diluted extracts. Method 1609 failed in only 11% and 3% of undiluted and diluted extracts analyses. Use of sample processing control assay results in the delta-delta Ct method for calculating relative target gene recoveries increased the number of acceptable recovery results. Delta-delta tended to bias recoveries fr

Comparison of Agar Dilution, Disk Diffusion, MicroScan, and Vitek Antimicrobial Susceptibility Testing Methods to Broth Microdilution for Detection of Fluoroquinolone-Resistant Isolates of the Family Enterobacteriaceae

PubMed Central

Steward, Christine D.; Stocker, Sheila A.; Swenson, Jana M.; O’Hara, Caroline M.; Edwards, Jonathan R.; Gaynes, Robert P.; McGowan, John E.; Tenover, Fred C.

1999-01-01

Fluoroquinolone resistance appears to be increasing in many species of bacteria, particularly in those causing nosocomial infections. However, the accuracy of some antimicrobial susceptibility testing methods for detecting fluoroquinolone resistance remains uncertain. Therefore, we compared the accuracy of the results of agar dilution, disk diffusion, MicroScan Walk Away Neg Combo 15 conventional panels, and Vitek GNS-F7 cards to the accuracy of the results of the broth microdilution reference method for detection of ciprofloxacin and ofloxacin resistance in 195 clinical isolates of the family Enterobacteriaceae collected from six U.S. hospitals for a national surveillance project (Project ICARE [Intensive Care Antimicrobial Resistance Epidemiology]). For ciprofloxacin, very major error rates were 0% (disk diffusion and MicroScan), 0.9% (agar dilution), and 2.7% (Vitek), while major error rates ranged from 0% (agar dilution) to 3.7% (MicroScan and Vitek). Minor error rates ranged from 12.3% (agar dilution) to 20.5% (MicroScan). For ofloxacin, no very major errors were observed, and major errors were noted only with MicroScan (3.7% major error rate). Minor error rates ranged from 8.2% (agar dilution) to 18.5% (Vitek). Minor errors for all methods were substantially reduced when results with MICs within ±1 dilution of the broth microdilution reference MIC were excluded from analysis. However, the high number of minor errors by all test systems remains a concern. PMID:9986809

Novel method for on-road emission factor measurements using a plume capture trailer.

PubMed

Morawska, L; Ristovski, Z D; Johnson, G R; Jayaratne, E R; Mengersen, K

2007-01-15

The method outlined provides for emission factor measurements to be made for unmodified vehicles driving under real world conditions at minimal cost. The method consists of a plume capture trailer towed behind a test vehicle. The trailer collects a sample of the naturally diluted plume in a 200 L conductive bag and this is delivered immediately to a mobile laboratory for subsequent analysis of particulate and gaseous emissions. The method offers low test turnaround times with the potential to complete much larger numbers of emission factor measurements than have been possible using dynamometer testing. Samples can be collected at distances up to 3 m from the exhaust pipe allowing investigation of early dilution processes. Particle size distribution measurements, as well as particle number and mass emission factor measurements, based on naturally diluted plumes are presented. A dilution profile relating the plume dilution ratio to distance from the vehicle tail pipe for a diesel passenger vehicle is also presented. Such profiles are an essential input for new mechanistic roadway air quality models.

Synthesis and formation mechanism of pinnoite in sulfated-type boron concentrated brine by dilution method

NASA Astrophysics Data System (ADS)

Peng, Jiaoyu; Bian, Shaoju; Lin, Feng; Wang, Liping; Dong, Yaping; Li, Wu

2017-10-01

The synthesis of pinnoite (MgB2O(OH)6) in boron-containing brine was established with a novel dilution method. Effects of temperature, precipitation time, boron concentration and mass dilution ratio on the formation of pinnoite were investigated. The products obtained were characterized by X-ray diffraction (XRD), Raman, thermogravimetric and differential scanning calorimeter (TG-DSC), and scanning electron microscopy. The transformation mechanism of pinnoite with different dilution ratios was assumed by studying the crystal growth of pinnoite. The results showed that pinnoite was synthesized above 60 °C in the diluted brine. There were two reaction steps - precipitation of amorphous solid and the formation of pinnoite crystals - during the whole reaction process of pinnoite when the dilution ratio is more than 1.0 at 80 °C. While in the 0.5 diluted brine, only one reaction step of pinnoite crystal formation was observed and its transformation mechanism was discussed based on dissociation of polyborates in brine. Besides, the origin of pinnoite mineral deposited on salt lake bottom was proposed.

21 CFR 175.270 - Poly(vinyl fluoride) resins.

Code of Federal Regulations, 2014 CFR

2014-04-01

...(vinyl fluoride) basic resins have an intrinsic viscosity of not less than 0.75 deciliter per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution Viscosity of Vinyl Chloride... (ASTM method D1243-79, “Standard Test Method for Dilute Solution Viscosity of Vinyl Chloride Polymers...

Assessing the Accuracy of the Tracer Dilution Method with Atmospheric Dispersion Modeling

NASA Astrophysics Data System (ADS)

Taylor, D.; Delkash, M.; Chow, F. K.; Imhoff, P. T.

2015-12-01

Landfill methane emissions are difficult to estimate due to limited observations and data uncertainty. The mobile tracer dilution method is a widely used and cost-effective approach for predicting landfill methane emissions. The method uses a tracer gas released on the surface of the landfill and measures the concentrations of both methane and the tracer gas downwind. Mobile measurements are conducted with a gas analyzer mounted on a vehicle to capture transects of both gas plumes. The idea behind the method is that if the measurements are performed far enough downwind, the methane plume from the large area source of the landfill and the tracer plume from a small number of point sources will be sufficiently well-mixed to behave similarly, and the ratio between the concentrations will be a good estimate of the ratio between the two emissions rates. The mobile tracer dilution method is sensitive to different factors of the setup such as placement of the tracer release locations and distance from the landfill to the downwind measurements, which have not been thoroughly examined. In this study, numerical modeling is used as an alternative to field measurements to study the sensitivity of the tracer dilution method and provide estimates of measurement accuracy. Using topography and wind conditions for an actual landfill, a landfill emissions rate is prescribed in the model and compared against the emissions rate predicted by application of the tracer dilution method. Two different methane emissions scenarios are simulated: homogeneous emissions over the entire surface of the landfill, and heterogeneous emissions with a hot spot containing 80% of the total emissions where the daily cover area is located. Numerical modeling of the tracer dilution method is a useful tool for evaluating the method without having the expense and labor commitment of multiple field campaigns. Factors tested include number of tracers, distance between tracers, distance from landfill to transect path, and location of tracers with respect to the hot spot. Results show that location of the tracers relative to the hot spot of highest landfill emissions makes the largest difference in accuracy of the tracer dilution method.

Particles size distribution in diluted magnetic fluids

NASA Astrophysics Data System (ADS)

Yerin, Constantine V.

2017-06-01

Changes in particles and aggregates size distribution in diluted kerosene based magnetic fluids is studied by dynamic light scattering method. It has been found that immediately after dilution in magnetic fluids the system of aggregates with sizes ranging from 100 to 250-1000 nm is formed. In 50-100 h after dilution large aggregates are peptized and in the sample stationary particles and aggregates size distribution is fixed.

Comparison of BOD results obtained by dilution and manometric methods in sanitary landfill leachates.

PubMed

Ceçen, F; Yangin, C

2000-12-01

This study examined the determination of BOD in landfill leachates by dilution (D-method) and manometric methods (M-method). The differences in results were discussed based on statistical tests. The effects of sample dilution, seeding, chloride and total Kjeldahl nitrogen (TKN) level were examined. The M-method was found to be more sensitive to increases in chloride and TKN concentrations. However, in the M-method the positive interference of nitrogenous BOD (NBOD) to carbonaceous BOD (CBOD) was more successfully prevented. The BOD rate constant k and the ultimate BOD (BODu) were estimated by non-linear regression. With the M-method these parameters could be more reliably estimated than the D-method. Suggestions were made for BOD analyses in landfill leachates in future studies.

Optimal back-extrapolation method for estimating plasma volume in humans using the indocyanine green dilution method.

PubMed

Polidori, David; Rowley, Clarence

2014-07-22

The indocyanine green dilution method is one of the methods available to estimate plasma volume, although some researchers have questioned the accuracy of this method. We developed a new, physiologically based mathematical model of indocyanine green kinetics that more accurately represents indocyanine green kinetics during the first few minutes postinjection than what is assumed when using the traditional mono-exponential back-extrapolation method. The mathematical model is used to develop an optimal back-extrapolation method for estimating plasma volume based on simulated indocyanine green kinetics obtained from the physiological model. Results from a clinical study using the indocyanine green dilution method in 36 subjects with type 2 diabetes indicate that the estimated plasma volumes are considerably lower when using the traditional back-extrapolation method than when using the proposed back-extrapolation method (mean (standard deviation) plasma volume = 26.8 (5.4) mL/kg for the traditional method vs 35.1 (7.0) mL/kg for the proposed method). The results obtained using the proposed method are more consistent with previously reported plasma volume values. Based on the more physiological representation of indocyanine green kinetics and greater consistency with previously reported plasma volume values, the new back-extrapolation method is proposed for use when estimating plasma volume using the indocyanine green dilution method.

Using the tracer-dilution discharge method to develop streamflow records for ice-affected streams in Colorado

USGS Publications Warehouse

Capesius, Joseph P.; Sullivan, Joseph R.; O'Neill, Gregory B.; Williams, Cory A.

2005-01-01

Accurate ice-affected streamflow records are difficult to obtain for several reasons, which makes the management of instream-flow water rights in the wintertime a challenging endeavor. This report documents a method to improve ice-affected streamflow records for two gaging stations in Colorado. In January and February 2002, the U.S. Geological Survey, in cooperation with the Colorado Water Conservation Board, conducted an experiment using a sodium chloride tracer to measure streamflow under ice cover by the tracer-dilution discharge method. The purpose of this study was to determine the feasibility of obtaining accurate ice-affected streamflow records by using a sodium chloride tracer that was injected into the stream. The tracer was injected at two gaging stations once per day for approximately 20 minutes for 25 days. Multiple-parameter water-quality sensors at the two gaging stations monitored background and peak chloride concentrations. These data were used to determine discharge at each site. A comparison of the current-meter streamflow record to the tracer-dilution streamflow record shows different levels of accuracy and precision of the tracer-dilution streamflow record at the two sites. At the lower elevation and warmer site, Brandon Ditch near Whitewater, the tracer-dilution method overestimated flow by an average of 14 percent, but this average is strongly biased by outliers. At the higher elevation and colder site, Keystone Gulch near Dillon, the tracer-dilution method experienced problems with the tracer solution partially freezing in the injection line. The partial freezing of the tracer contributed to the tracer-dilution method underestimating flow by 52 percent at Keystone Gulch. In addition, a tracer-pump-reliability test was conducted to test how accurately the tracer pumps can discharge the tracer solution in conditions similar to those used at the gaging stations. Although the pumps were reliable and consistent throughout the 25-day study period, the pumps underdischarged the tracer by 5.8-15.9 percent as compared to the initial pumping rate setting, which may explain some of the error in the tracer-dilution streamflow record as compared to current-meter streamflow record.

Creatine Supplementation Increases Total Body Water in Soccer Players: a Deuterium Oxide Dilution Study.

PubMed

Deminice, R; Rosa, F T; Pfrimer, K; Ferrioli, E; Jordao, A A; Freitas, E

2016-02-01

This study aimed to evaluate changes in total body water (TBW) in soccer athletes using a deuterium oxide dilution method and bioelectrical impedance (BIA) formulas after 7 days of creatine supplementation. In a double-blind controlled manner, 13 healthy (under-20) soccer players were divided randomly in 2 supplementation groups: Placebo (Pla, n=6) and creatine supplementation (CR, n=7). Before and after the supplementation period (0.3 g/kg/d during 7 days), TBW was determined by deuterium oxide dilution and BIA methods. 7 days of creatine supplementation lead to a large increase in TBW (2.3±1.0 L) determined by deuterium oxide dilution, and a small but significant increase in total body weight (1.0±0.4 kg) in Cr group compared to Pla. The Pla group did not experience any significant changes in TBW or body weight. Although 5 of 6 BIA equations were sensitive to determine TBW changes induced by creatine supplementation, the Kushner et al. 16 method presented the best concordance levels when compared to deuterium dilution method. In conclusion, 7-days of creatine supplementation increased TBW determined by deuterium oxide dilution or BIA formulas. BIA can be useful to determine TBW changes promoted by creatine supplementation in soccer athletes, with special concern for formula choice. © Georg Thieme Verlag KG Stuttgart · New York.

Biodiesel Impact on Engine Lubricant Dilution During Active Regeneration of Aftertreatment Systems

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

He, X.; Williams, A.; Christensen, E.

Experiments were conducted with ultra low sulfur diesel (ULSD) and 20% biodiesel blends (B20) to compare lube oil dilution levels and lubricant properties for systems using late in-cylinder fuel injection for aftertreatment regeneration. Lube oil dilution was measured by gas chromatography (GC) following ASTM method D3524 to measure diesel content, by Fourier transform infrared (FTIR) spectrometry following a modified ASTM method D7371 to measure biodiesel content, and by a newly developed back-flush GC method that simultaneously measures both diesel and biodiesel. Heavy-duty (HD) engine testing was conducted on a 2008 6.7L Cummins ISB equipped with a diesel oxidation catalyst (DOC)more » and diesel particle filter (DPF). Stage one of engine testing consisted of 10 consecutive repeats of a forced DPF regeneration event. This continuous operation with late in-cylinder fuel injection served as a method to accelerate lube-oil dilution. Stage two consisted of 16 hours of normal engine operation over a transient test cycle, which created an opportunity for any accumulated fuel in the oil sump to evaporate. Light duty (LD) vehicle testing was conducted on a 2010 VW Jetta equipped with DOC, DPF and a NOx storage catalyst (NSC). Vehicle testing comprised approximately 4,000 miles of operation on a mileage-accumulation dynamometer (MAD) using the U.S. Environmental Protection Agency's Highway Fuel Economy Cycle because of the relatively low engine oil and exhaust temperatures, and high DPF regeneration frequency of this cycle relative to other cycles examined. Comparison of the lube oil dilution analysis methods suggests that D3524 does not measure dilution by biodiesel. The new back-flush GC method provided analysis for both diesel and biodiesel, in a shorter time and with lower detection limit. Thus all lube oil dilution results in this paper are based on this method. Analysis of the HD lube-oil samples showed only 1.5% to 1.6% fuel dilution for both fuels during continuous operation under DPF regeneration events. During the second stage of HD testing, the ULSD lube-oil dilution levels fell from 1.5% to 0.8%, while for B20, lube-oil dilution levels fell from 1.6% to 1.0%, but the fuel in the oil was 36% biodiesel. For the LD vehicle tests, the frequency of DPF regeneration events was observed to be the same for both ULSD and B20. No significant difference between the two fuels' estimated soot loading was detected by the engine control unit (ECU), although a 23% slower rate of increase in differential pressure across DPF was observed with B20. It appears that the ECU estimated soot loading is based on the engine map, not taking advantage of the lower engine-out particulate matter from the use of biodiesel. After 4,000 miles of LD vehicle operation with ULSD, fuel dilution in the lube-oil samples showed total dilution levels of 4.1% diesel. After 4,000 miles of operation with B20, total fuel in oil dilution levels were 6.7% consisting of 3.6% diesel fuel and 3.1% biodiesel. Extrapolation to the 10,000-mile oil drain interval with B20 suggests that the total fuel content in the oil could reach 12%, compared to 5% for operation on ULSD. Analysis of the oil samples also included measurement of total acid number, total base number, viscosity, soot, metals and wear scar; however, little difference in these parameters was noted.« less

Numerical simulations to assess the tracer dilution method for measurement of landfill methane emissions.

PubMed

Taylor, Diane M; Chow, Fotini K; Delkash, Madjid; Imhoff, Paul T

2016-10-01

Landfills are a significant contributor to anthropogenic methane emissions, but measuring these emissions can be challenging. This work uses numerical simulations to assess the accuracy of the tracer dilution method, which is used to estimate landfill emissions. Atmospheric dispersion simulations with the Weather Research and Forecast model (WRF) are run over Sandtown Landfill in Delaware, USA, using observation data to validate the meteorological model output. A steady landfill methane emissions rate is used in the model, and methane and tracer gas concentrations are collected along various transects downwind from the landfill for use in the tracer dilution method. The calculated methane emissions are compared to the methane emissions rate used in the model to find the percent error of the tracer dilution method for each simulation. The roles of different factors are examined: measurement distance from the landfill, transect angle relative to the wind direction, speed of the transect vehicle, tracer placement relative to the hot spot of methane emissions, complexity of topography, and wind direction. Results show that percent error generally decreases with distance from the landfill, where the tracer and methane plumes become well mixed. Tracer placement has the largest effect on percent error, and topography and wind direction both have significant effects, with measurement errors ranging from -12% to 42% over all simulations. Transect angle and transect speed have small to negligible effects on the accuracy of the tracer dilution method. These tracer dilution method simulations provide insight into measurement errors that might occur in the field, enhance understanding of the method's limitations, and aid interpretation of field data. Copyright © 2016 Elsevier Ltd. All rights reserved.

Congener-specific determination of ultratrace levels of chlorinated and brominated polycyclic aromatic hydrocarbons in atmosphere and industrial stack gas by isotopic dilution gas chromatography/high resolution mass spectrometry method.

PubMed

Jin, Rong; Liu, Guorui; Zheng, Minghui; Fiedler, Heidelore; Jiang, Xiaoxu; Yang, Lili; Wu, Xiaolin; Xu, Yang

2017-08-04

Isotopic dilution gas chromatography combined with high resolution mass spectrometry (GC/HRMS) has overwhelming advantages with respect to the accuracy of congener-specific ultratrace analysis of complex persistent organic pollutants (POPs) in environmental matrices. However, an isotopic dilution GC/HRMS method for analysis of chlorinated and brominated polycyclic aromatic hydrocarbons (Cl-PAHs and Br-PAHs) using 13 C-labelled congeners as internal standards has not been established. In this study, a method for identification and quantification of 38 congeners of Cl-PAHs and Br-PAHs in atmosphere and stack gas samples from waste incinerators was developed using the isotopic dilution GC/HRMS technique. The instrumental detection limits of the GC/HRMS method ranged from 0.2pg to 1.8pg for Cl-PAH congeners, and 0.7pg to 2.7pg for Br-PAH congeners, which were about three orders of magnitude lower than those of the GC/quadrupole MS method. This new method developed was also the first to enable determination of Cl-PAH and Br-PAH homologs comprising congeners with the same molecular skeleton and chlorine or bromine substitution numbers. Among the detected congeners, seven Cl-PAH congeners and thirteen Br-PAH congeners that were abundant in the atmosphere and stack gases released from waste incinerators were firstly detected in real samples and reported using the established isotopic dilution GC/HRMS method. The developed isotopic dilution GC/HRMS is significant and needed for better studying the environmental behavior and health risk of Cl-PAHs and Br-PAHs. Copyright © 2017 Elsevier B.V. All rights reserved.

Antibacterial activities of naturally occurring compounds against Mycobacterium avium subsp. paratuberculosis.

PubMed

Wong, Stella Y Y; Grant, Irene R; Friedman, Mendel; Elliott, Christopher T; Situ, Chen

2008-10-01

The antibacterial activities of 18 naturally occurring compounds (including essential oils and some of their isolated constituents, apple and green tea polyphenols, and other plant extracts) against three strains of Mycobacterium avium subsp. paratuberculosis (a bovine isolate [NCTC 8578], a raw-milk isolate [806R], and a human isolate [ATCC 43015]) were evaluated using a macrobroth susceptibility testing method. M. avium subsp. paratuberculosis was grown in 4 ml Middlebrook 7H9 broth containing 10% oleic acid-albumin-dextrose-catalase, 0.05% Tween 80 (or 0.2% glycerol), and 2 microg/ml mycobactin J supplemented with five concentrations of each test compound. The changes in the optical densities of the cultures at 600 nm as a measure of CFU were recorded at intervals over an incubation period of 42 days at 37 degrees C. Six of the compounds were found to inhibit the growth of M. avium subsp. paratuberculosis. The most effective compound was trans-cinnamaldehyde, with a MIC of 25.9 microg/ml, followed by cinnamon oil (26.2 microg/ml), oregano oil (68.2 microg/ml), carvacrol (72.2 microg/ml), 2,5-dihydroxybenzaldehyde (74 microg/ml), and 2-hydroxy-5-methoxybenzaldehyde (90.4 microg/ml). With the exception of carvacrol, a phenolic compound, three of the four most active compounds are aldehydes, suggesting that the structure of the phenolic group or the aldehyde group may be important to the antibacterial activity. No difference in compound activity was observed between the three M. avium subsp. paratuberculosis strains studied. Possible mechanisms of the antimicrobial effects are discussed.

Antibacterial Activities of Naturally Occurring Compounds against Mycobacterium avium subsp. paratuberculosis▿

PubMed Central

Wong, Stella Y. Y.; Grant, Irene R.; Friedman, Mendel; Elliott, Christopher T.; Situ, Chen

2008-01-01

The antibacterial activities of 18 naturally occurring compounds (including essential oils and some of their isolated constituents, apple and green tea polyphenols, and other plant extracts) against three strains of Mycobacterium avium subsp. paratuberculosis (a bovine isolate [NCTC 8578], a raw-milk isolate [806R], and a human isolate [ATCC 43015]) were evaluated using a macrobroth susceptibility testing method. M. avium subsp. paratuberculosis was grown in 4 ml Middlebrook 7H9 broth containing 10% oleic acid-albumin-dextrose-catalase, 0.05% Tween 80 (or 0.2% glycerol), and 2 μg/ml mycobactin J supplemented with five concentrations of each test compound. The changes in the optical densities of the cultures at 600 nm as a measure of CFU were recorded at intervals over an incubation period of 42 days at 37°C. Six of the compounds were found to inhibit the growth of M. avium subsp. paratuberculosis. The most effective compound was trans-cinnamaldehyde, with a MIC of 25.9 μg/ml, followed by cinnamon oil (26.2 μg/ml), oregano oil (68.2 μg/ml), carvacrol (72.2 μg/ml), 2,5-dihydroxybenzaldehyde (74 μg/ml), and 2-hydroxy-5-methoxybenzaldehyde (90.4 μg/ml). With the exception of carvacrol, a phenolic compound, three of the four most active compounds are aldehydes, suggesting that the structure of the phenolic group or the aldehyde group may be important to the antibacterial activity. No difference in compound activity was observed between the three M. avium subsp. paratuberculosis strains studied. Possible mechanisms of the antimicrobial effects are discussed. PMID:18676709

Methods for deoxygenating biomass-derived pyrolysis oil

DOEpatents

Baird, Lance Awender; Brandvold, Timothy A.

2015-06-30

Methods for deoxygenating a biomass-derived pyrolysis oil are provided. A method for deoxygenating a biomass-derived pyrolysis oil comprising the steps of combining a biomass-derived pyrolysis oil stream with a heated low-oxygen-pyoil diluent recycle stream to form a heated diluted pyoil feed stream is provided. The heated diluted pyoil feed stream has a feed temperature of about 150.degree. C. or greater. The heated diluted pyoil feed stream is contacted with a first deoxygenating catalyst in the presence of hydrogen at first hydroprocessing conditions effective to form a low-oxygen biomass-derived pyrolysis oil effluent.

Detection of Toxoplasma oocysts from soil by modified sucrose flotation and PCR methods.

PubMed

Matsuo, Junji; Kimura, Daisuke; Rai, Shiba Kumar; Uga, Shoji

2004-06-01

A detection method of Toxoplasma gondii oocysts from soil was evaluated using the sucrose flotation technique with modification involving addition of 0.1% gelatin into washing and floating solutions. PCR was performed on untreated samples and after treatment with polyvinylpyrrolidone (PVP), heating and cooling, and NaCl. The addition of gelatin in the sucrose solution yielded a higher number of oocysts. A very thin band was observed when DNA extract was diluted to 1:1024, indicating the presence of PCR inhibitor in the soil. PCR performed on untreated DNA, on PVP-treated, and on PVP-treated with heating and cooling without added bovine serum albumin (BSA) showed a band only at higher dilutions (1:1024 and 1:512) but at a much lower dilution (1:8) with BSA. In contrast, DNA treated with all three agents showed a band at a much lower dilution (1:64), even without added BSA, and no dilution was required when BSA was added. The PCR inhibitors present in the soil were removed by employing various treatment procedures during DNA extraction, and BSA in PCR. Furthermore, the detection limit with the method was 1 oocyst/g of soil, indicating that this method is useful in epidemiological studies.

A method for predicting optimized processing parameters for surfacing

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

Dupont, J.N.; Marder, A.R.

1994-12-31

Welding is used extensively for surfacing applications. To operate a surfacing process efficiently, the variables must be optimized to produce low levels of dilution with the substrate while maintaining high deposition rates. An equation for dilution in terms of the welding variables, thermal efficiency factors, and thermophysical properties of the overlay and substrate was developed by balancing energy and mass terms across the welding arc. To test the validity of the resultant dilution equation, the PAW, GTAW, GMAW, and SAW processes were used to deposit austenitic stainless steel onto carbon steel over a wide range of parameters. Arc efficiency measurementsmore » were conducted using a Seebeck arc welding calorimeter. Melting efficiency was determined based on knowledge of the arc efficiency. Dilution was determined for each set of processing parameters using a quantitative image analysis system. The pertinent equations indicate dilution is a function of arc power (corrected for arc efficiency), filler metal feed rate, melting efficiency, and thermophysical properties of the overlay and substrate. With the aid of the dilution equation, the effect of processing parameters on dilution is presented by a new processing diagram. A new method is proposed for determining dilution from welding variables. Dilution is shown to depend on the arc power, filler metal feed rate, arc and melting efficiency, and the thermophysical properties of the overlay and substrate. Calculated dilution levels were compared with measured values over a large range of processing parameters and good agreement was obtained. The results have been applied to generate a processing diagram which can be used to: (1) predict the maximum deposition rate for a given arc power while maintaining adequate fusion with the substrate, and (2) predict the resultant level of dilution with the substrate.« less

Single-molecule dilution and multiple displacement amplification for molecular haplotyping.

PubMed

Paul, Philip; Apgar, Josh

2005-04-01

Separate haploid analysis is frequently required for heterozygous genotyping to resolve phase ambiguity or confirm allelic sequence. We demonstrate a technique of single-molecule dilution followed by multiple strand displacement amplification to haplotype polymorphic alleles. Dilution of DNA to haploid equivalency, or a single molecule, is a simple method for separating di-allelic DNA. Strand displacement amplification is a robust method for non-specific DNA expansion that employs random hexamers and phage polymerase Phi29 for double-stranded DNA displacement and primer extension, resulting in high processivity and exceptional product length. Single-molecule dilution was followed by strand displacement amplification to expand separated alleles to microgram quantities of DNA for more efficient haplotype analysis of heterozygous genes.

Approaches of aroma extraction dilution analysis (AEDA) for headspace solid phase microextraction and gas chromatography-olfactometry (HS-SPME-GC-O): Altering sample amount, diluting the sample or adjusting split ratio?

PubMed

Feng, Yunzi; Cai, Yu; Sun-Waterhouse, Dongxiao; Cui, Chun; Su, Guowan; Lin, Lianzhu; Zhao, Mouming

2015-11-15

Aroma extract dilution analysis (AEDA) is widely used for the screening of aroma-active compounds in gas chromatography-olfactometry (GC-O). In this study, three aroma dilution methods, (I) using different test sample volumes, (II) diluting samples, and (III) adjusting the GC injector split ratio, were compared for the analysis of volatiles by using HS-SPME-AEDA. Results showed that adjusting the GC injector split ratio (III) was the most desirable approach, based on the linearity relationships between Ln (normalised peak area) and Ln (normalised flavour dilution factors). Thereafter this dilution method was applied in the analysis of aroma-active compounds in Japanese soy sauce and 36 key odorants were found in this study. The most intense aroma-active components in Japanese soy sauce were: ethyl 2-methylpropanoate, ethyl 2-methylbutanoate, ethyl 3-methylbutanoate, ethyl 4-methylpentanoate, 3-(methylthio)propanal, 1-octen-3-ol, 2-methoxyphenol, 4-ethyl-2-methoxyphenol, 2-methoxy-4-vinylphenol, 2-phenylethanol, and 4-hydroxy-5-ethyl-2-methyl-3(2H)-furanone. Copyright © 2015. Published by Elsevier Ltd.

Simplified Method for Groundwater Treatment Using Dilution and Ceramic Filter

NASA Astrophysics Data System (ADS)

Musa, S.; Ariff, N. A.; Kadir, M. N. Abdul; Denan, F.

2016-07-01

Groundwater is one of the natural resources that is not susceptible to pollutants. However, increasing activities of municipal, industrial, agricultural or extreme land use activities have resulted in groundwater contamination as occured at the Research Centre for Soft Soil Malaysia (RECESS), Universiti Tun Hussein Onn Malaysia (UTHM). Thus, aims of this study is to treat groundwater by using rainwater and simple ceramic filter as a treatment agent. The treatment uses rain water dilution, ceramic filters and combined method of dilute and filtering as an alternate treatment which are simple and more practical compared to modern or chemical methods. The water went through dilution treatment processes able to get rid of 57% reduction compared to initial condition. Meanwhile, the water that passes through the filtering process successfully get rid of as much as 86% groundwater parameters where only chloride does not pass the standard. Favorable results for the combination methods of dilution and filtration methods that can succesfully eliminate 100% parameters that donot pass the standards of the Ministry of Health and the Interim National Drinking Water Quality Standard such as those found in groundwater in RECESS, UTHM especially sulfate and chloride. As a result, it allows the raw water that will use clean drinking water and safe. It also proves that the method used in this study is very effective in improving the quality of groundwater.

Measurement of plasma unbound unconjugated bilirubin.

PubMed

Ahlfors, C E

2000-03-15

A method is described for measuring the unconjugated fraction of the unbound bilirubin concentration in plasma by combining the peroxidase method for determining unbound bilirubin with a diazo method for measuring conjugated and unconjugated bilirubin. The accuracy of the unbound bilirubin determination is improved by decreasing sample dilution, eliminating interference by conjugated bilirubin, monitoring changes in bilirubin concentration using diazo derivatives, and correcting for rate-limiting dissociation of bilirubin from albumin. The unbound unconjugated bilirubin concentration by the combined method in plasma from 20 jaundiced newborns was significantly greater than and poorly correlated with the unbound bilirubin determined by the existing peroxidase method (r = 0.7), possibly due to differences in sample dilution between the methods. The unbound unconjugated bilirubin was an unpredictable fraction of the unbound bilirubin in plasma samples from patients with similar total bilirubin concentrations but varying levels of conjugated bilirubin. A bilirubin-binding competitor was readily detected at a sample dilution typically used for the combined test but not at the dilution used for the existing peroxidase method. The combined method is ideally suited to measuring unbound unconjugated bilirubin in jaundiced human newborns or animal models of kernicterus. Copyright 2000 Academic Press.

New methods allowing the detection of protein aggregates

PubMed Central

Demeule, Barthélemy; Palais, Caroline; Machaidze, Gia; Gurny, Robert

2009-01-01

Aggregation compromises the safety and efficacy of therapeutic proteins. According to the manufacturer, the therapeutic immunoglobulin trastuzumab (Herceptin®) should be diluted in 0.9% sodium chloride before administration. Dilution in 5% dextrose solutions is prohibited. The reason for the interdiction is not mentioned in the Food and Drug Administration (FDA) documentation, but the European Medicines Agency (EMEA) Summary of Product Characteristics states that dilution of trastuzumab in dextrose solutions results in protein aggregation. In this paper, asymmetrical flow field-flow fractionation (FFF), fluorescence spectroscopy, fluorescence microscopy and transmission electron microscopy (TEM) have been used to characterize trastuzumab samples diluted in 0.9% sodium chloride, a stable infusion solution, as well as in 5% dextrose (a solution prone to aggregation). When trastuzumab samples were injected in the FFF channel using a standard separation method, no difference could be seen between trastuzumab diluted in sodium chloride and trastuzumab diluted in dextrose. However, during FFF measurements made with appropriate protocols, aggregates were detected in 5% dextrose. The parameters enabling the detection of reversible trastuzumab aggregates are described. Aggregates could also be documented by fluorescence microscopy and TEM. Fluorescence spectroscopy data were indicative of conformational changes consistent with increased aggregation and adsorption to surfaces. The analytical methods presented in this study were able to detect and characterize trastuzumab aggregates. PMID:20061815

Optimal back-extrapolation method for estimating plasma volume in humans using the indocyanine green dilution method

PubMed Central

2014-01-01

Background The indocyanine green dilution method is one of the methods available to estimate plasma volume, although some researchers have questioned the accuracy of this method. Methods We developed a new, physiologically based mathematical model of indocyanine green kinetics that more accurately represents indocyanine green kinetics during the first few minutes postinjection than what is assumed when using the traditional mono-exponential back-extrapolation method. The mathematical model is used to develop an optimal back-extrapolation method for estimating plasma volume based on simulated indocyanine green kinetics obtained from the physiological model. Results Results from a clinical study using the indocyanine green dilution method in 36 subjects with type 2 diabetes indicate that the estimated plasma volumes are considerably lower when using the traditional back-extrapolation method than when using the proposed back-extrapolation method (mean (standard deviation) plasma volume = 26.8 (5.4) mL/kg for the traditional method vs 35.1 (7.0) mL/kg for the proposed method). The results obtained using the proposed method are more consistent with previously reported plasma volume values. Conclusions Based on the more physiological representation of indocyanine green kinetics and greater consistency with previously reported plasma volume values, the new back-extrapolation method is proposed for use when estimating plasma volume using the indocyanine green dilution method. PMID:25052018

The effect of glycerol-related osmotic changes on post-thaw motility and acrosomal integrity of ram spermatozoa.

PubMed

Fiser, P S; Fairfull, R W

1989-02-01

Ram semen, collected by artificial vagina, was diluted and processed for long-term storage as described by P. S. Fiser, L. Ainsworth, and R. W. Fairfull (Canad. J. Anim. Sci. 62, 425-428, 1982). The concentration of the cryoprotectant, glycerol, was adjusted to 4% in the diluted semen prior to freezing by a one-step addition at 30 degrees C (Method 1), by cooling the semen to 5 degrees C and addition of the glycerol gradually over 30 min (Method 2), by one-step addition of glycerol prior to equilibration for 2 hr (Method 3), or by cooling to 5 degrees C, followed by a holding period of 2 hr at 5 degrees C, and the one-step addition of glycerol just prior to freezing (Method 4). After thawing, the glycerol concentration of the semen was reduced by stepwise dilution from 4 to 0.4% over 15 or 30 min or by a one-step ten-fold dilution. The average post-thaw percentage of motile spermatozoa was significantly lower after addition of glycerol by Method 1 (39.9%) than when the glycerol was added by the other three methods (range, 44.0-46.4% averaged over the glycerol dilution). The average post-thaw percentage of intact acrosomes (61.2%), highest in semen in which the glycerol was added by Method 2, was not significantly different from those in which glycerol was added to semen by Methods 3 and 4, but it was significantly higher than that found in semen in which the glycerol was added by Method 1 (54.4%). However, when averaged over the method of glycerolation, the post-thaw percentage of motile spermatozoa (range, 43.7-44.2%) and the percentage of intact acrosomes (range, 56.8-59.5%) did not differ significantly in semen subjected to gradual decrease in glycerol concentration and diluent osmolality (over 15 and 30 min) or by a one-step, 10-fold dilution. These data indicate that post-thaw survival of spermatozoa can be influenced by the way in which glycerol is added prior to freezing. However, post-thaw spermatozoa motility and acrosomal integrity can be maintained even after a rapid decrease in glycerol concentration such as that which accompanies insemination or dilution of semen for assessment of motility.

Doping control analysis of 46 polar drugs in horse plasma and urine using a 'dilute-and-shoot' ultra high performance liquid chromatography-high resolution mass spectrometry approach.

PubMed

Kwok, Wai Him; Choi, Timmy L S; Kwok, Karen Y; Chan, George H M; Wong, Jenny K Y; Wan, Terence S M

2016-06-17

The high sensitivity of ultra high performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS) allows the identification of many prohibited substances without pre-concentration, leading to the development of simple and fast 'dilute-and-shoot' methods for doping control for human and equine sports. While the detection of polar drugs in plasma and urine is difficult using liquid-liquid or solid-phase extraction as these substances are poorly extracted, the 'dilute-and-shoot' approach is plausible. This paper describes a 'dilute-and-shoot' UHPLC-HRMS screening method to detect 46 polar drugs in equine urine and plasma, including some angiotensin-converting enzyme (ACE) inhibitors, sympathomimetics, anti-epileptics, hemostatics, the new doping agent 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR), as well as two threshold substances, namely dimethyl sulfoxide and theobromine. For plasma, the sample (200μL) was protein precipitated using trichloroacetic acid, and the resulting supernatant was diluted using Buffer A with an overall dilution factor of 3. For urine, the sample (20μL) was simply diluted 50-fold with Buffer A. The diluted plasma or urine sample was then analysed using a UHPLC-HRMS system in full-scan ESI mode. The assay was validated for qualitative identification purpose. This straightforward and reliable approach carried out in combination with other screening procedures has increased the efficiency of doping control analysis in the laboratory. Moreover, since the UHPLC-HRMS data were acquired in full-scan mode, the method could theoretically accommodate an unlimited number of existing and new doping agents, and would allow a retrospectively search for drugs that have not been targeted at the time of analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

'Dilute-and-shoot' triple parallel mass spectrometry method for analysis of vitamin D and triacylglycerols in dietary supplements

USDA-ARS?s Scientific Manuscript database

A method is demonstrated for analysis of vitamin D-fortified dietary supplements that eliminates virtually all chemical pretreatment prior to analysis, and is referred to as a ‘dilute and shoot’ method. Three mass spectrometers, in parallel, plus a UV detector, an evaporative light scattering detec...

High-throughput method to predict extrusion pressure of ceramic pastes.

PubMed

Cao, Kevin; Liu, Yang; Tucker, Christopher; Baumann, Michael; Grit, Grote; Lakso, Steven

2014-04-14

A new method was developed to measure the rheology of extrudable ceramic pastes using a Hamilton MicroLab Star liquid handler. The Hamilton instrument, normally used for high throughput liquid processing, was expanded to function as a low pressure capillary rheometer. Diluted ceramic pastes were forced through the modified pipettes, which produced pressure drop data that was converted to standard rheology data. A known ceramic paste containing cellulose ether was made and diluted to various concentrations in water. The most dilute paste samples were tested in the Hamilton instrument and the more typical, highly concentrated, ceramic paste were tested with a hydraulic ram extruder fitted with a capillary die and pressure measurement system. The rheology data from this study indicates that the dilute high throughput method using the Hamilton instrument correlates to, and can predict, the rheology of concentrated ceramic pastes normally used in ceramic extrusion production processes.

Apparatuses and methods for deoxygenating biomass-derived pyrolysis oil

DOEpatents

Kalnes, Tom N.

2015-12-29

Apparatuses and methods for deoxygenating a biomass-derived pyrolysis oil are provided herein. In one example, the method comprises of dividing a feedstock stream into first and second feedstock portions. The feedstock stream comprises the biomass-derived pyrolysis oil and has a temperature of about 60.degree. C. or less. The first feedstock portion is combined with a heated organic liquid stream to form a first heated diluted pyoil feed stream. The first heated diluted pyoil feed stream is contacted with a first deoxygenating catalyst in the presence of hydrogen to form an intermediate low-oxygen pyoil effluent. The second feedstock portion is combined with the intermediate low-oxygen pyoil effluent to form a second heated diluted pyoil feed stream. The second heated diluted pyoil feed stream is contacted with a second deoxygenating catalyst in the presence of hydrogen to form additional low-oxygen pyoil effluent.

A dilute-and-shoot flow-injection tandem mass spectrometry method for quantification of phenobarbital in urine.

PubMed

Alagandula, Ravali; Zhou, Xiang; Guo, Baochuan

2017-01-15

Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is the gold standard of urine drug testing. However, current LC-based methods are time consuming, limiting the throughput of MS-based testing and increasing the cost. This is particularly problematic for quantification of drugs such as phenobarbital, which is often analyzed in a separate run because they must be negatively ionized. This study examined the feasibility of using a dilute-and-shoot flow-injection method without LC separation to quantify drugs with phenobarbital as a model system. Briefly, a urine sample containing phenobarbital was first diluted by 10 times, followed by flow injection of the diluted sample to mass spectrometer. Quantification and detection of phenobarbital were achieved by an electrospray negative ionization MS/MS system operated in the multiple reaction monitoring (MRM) mode with the stable-isotope-labeled drug as internal standard. The dilute-and-shoot flow-injection method developed was linear with a dynamic range of 50-2000 ng/mL of phenobarbital and correlation coefficient > 0.9996. The coefficients of variation and relative errors for intra- and inter-assays at four quality control (QC) levels (50, 125, 445 and 1600 ng/mL) were 3.0% and 5.0%, respectively. The total run time to quantify one sample was 2 min, and the sensitivity and specificity of the method did not deteriorate even after 1200 consecutive injections. Our method can accurately and robustly quantify phenobarbital in urine without LC separation. Because of its 2 min run time, the method can process 720 samples per day. This feasibility study shows that the dilute-and-shoot flow-injection method can be a general way for fast analysis of drugs in urine. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Stable isotope dilution analysis of hydrologic samples by inductively coupled plasma mass spectrometry

USGS Publications Warehouse

Garbarino, John R.; Taylor, Howard E.

1987-01-01

Inductively coupled plasma mass spectrometry is employed in the determination of Ni, Cu, Sr, Cd, Ba, Ti, and Pb in nonsaline, natural water samples by stable isotope dilution analysis. Hydrologic samples were directly analyzed without any unusual pretreatment. Interference effects related to overlapping isobars, formation of metal oxide and multiply charged ions, and matrix composition were identified and suitable methods of correction evaluated. A comparability study snowed that single-element isotope dilution analysis was only marginally better than sequential multielement isotope dilution analysis. Accuracy and precision of the single-element method were determined on the basis of results obtained for standard reference materials. The instrumental technique was shown to be ideally suited for programs associated with certification of standard reference materials.

Methods for Measuring Lung Volumes: Is There a Better One?

PubMed

Tantucci, Claudio; Bottone, Damiano; Borghesi, Andrea; Guerini, Michele; Quadri, Federico; Pini, Laura

2016-01-01

Accurate measurement of lung volumes is of paramount importance to establish the presence of ventilatory defects and give insights for diagnostic and/or therapeutic purposes. It was the aim of this study to measure lung volumes in subjects with respiratory disorders and in normal controls by 3 different techniques (plethysmographic, dilutional and radiographic methods), in an attempt to clarify the role of each of them in performing such a task, without any presumptive 'a priori' superiority of one method above others. Patients andMethods: In different groups of subjects with obstructive and restrictive ventilatory defects and in a normal control group, total lung capacity, functional residual capacity (FRC) and residual volume were measured by body plethysmography, multi-breath helium (He) dilution and radiographic CT scan method with spirometric gating. The 3 methods gave comparable results in normal subjects and in patients with a restrictive defect. In patients with an obstructive defect, CT scan and plethysmography showed similar lung volumes, while on average significantly lower lung volumes were obtained with the He dilution technique. Taking into account that the He dilution technique does primarily measure FRC during tidal breathing, our data suggest that in some patients with an obstructive defect, a number of small airways can be functionally closed at end-expiratory lung volume, preventing He to reach the lung regions subserved by these airways. In all circumstances, both CT scan with spirometric gating and plethysmographic methods provide similar values of lung volumes. In contrast, the He dilution method can measure lower lung volumes in some patients with chronic airflow obstruction. © 2016 S. Karger AG, Basel.

Dynamics and estimates of growth and loss rates of bacterioplankton in a temperate freshwater system.

PubMed

Jugnia, Louis-B; Sime-Ngando, Télesphore; Gilbert, Daniel

2006-10-01

The growth rate and losses of bacterioplankton in the epilimnion of an oligo-mesotrophic reservoir were simultaneously estimated using three different methods for each process. Bacterial production was determined by means of the tritiated thymidine incorporation method, the dialysis bag method and the dilution method, while bacterial mortality was assessed with the dilution method, the disappearance of thymidine-labeled natural cells and ingestion of fluorescent bacterial tracers by heterotrophic flagellates. The different methods used to estimate bacterial growth rates yielded similar results. On the other hand, the mortality rates obtained with the dilution method were significantly lower than those obtained with the use of thymidine-labeled natural cells. The bacterial ingestion rate by flagellates accounted on average for 39% of total bacterial mortality estimated by the dilution method, but this value fell to 5% when the total mortality was measured by the thymidine-labeling method. Bacterial abundance and production varied in opposite phase to flagellate abundance and the various bacterial mortality rates. All this points to the critical importance of methodological aspects in the elaboration of quantitative models of matter and energy flows over the time through microbial trophic networks in aquatic systems, and highlights the role of bacterioplankton as a source of carbon for higher trophic levels in the studied system.

Impact of different dilution techniques on boar sperm quality and sperm distribution of the extended ejaculate.

PubMed

Schulze, M; Ammon, C; Schaefer, J; Luther, A-M; Jung, M; Waberski, D

2017-07-01

The dilution of ejaculates is a fundamental step for the production of liquid-preserved boar semen. For a long time, it has been recommended to add the extender to the ejaculate. The aim of the present study was to first compare the effect of the position ('center' vs. 'wall') where the extender is added to the semen-mixing cylinder (height 32.5cm; diameter 12.7cm) using an automatic dispenser (n=11). In experiment 2 (n=30), we analyzed the two main dilution methods (extender to the semen ('control') vs. 'reverse'). Experiment 3 was carried out to study the dilution effect on kinematics. In Experiments 1 and 2, the sperm distribution 10min after the dilution and the sperm quality parameters during long-term storage (d1, d3, d5, and d7) were evaluated. In Experiment 3, sperm quality was assessed during short-term storage at 0, 10, 20, 30 and 60min after semen dilution ('control' vs. 'reverse'; n=6). There were no significant differences (P>0.05) between the treatments in the specific response to bicarbonate, mitochondrial activity, membrane status, thermo-resistance or sperm motility immediately after dilution or long-term storage. The sperm distribution was significantly (P=0.029) affected by the dilution method in Experiment 2. In summary, treatment with the extender first, which is used by only a few European boar studs, leads to comparable results in sperm quality during storage and better results in sperm distribution after dilution. This procedure is also less time consuming, less foam formation occurs during the semen dilution and the procedure is more hygienic. Copyright © 2017 Elsevier B.V. All rights reserved.

Determination of the concentration of a Bryonia Dioica tincture by Raman spectroscopy

NASA Astrophysics Data System (ADS)

Milea, Irimie; Culea, E.; Iliescu, T.; Milea, Janetta

1995-03-01

It was established a method based on Raman spectroscopy in order to control the content and concentration of homeopathic dilutions. Dilutions of a tincture of Bryonia Dioica in ethanol were prepared and investigated by Raman spectroscopy. The Raman line at 881 cm-1 was found to depend linearly versus the concentration of Bryonia Dioica. This permits to obtain a calibration curve that may be used to determine the concentration of Bryonia Dioica in ethanol. The method may be extended to determine the concentration of various homeopathic dilutions.

Theoretical underpinning of the single-molecule-dilution (SMD) method of direct haplotype resolution.

PubMed Central

Stephens, J C; Rogers, J; Ruano, G

1990-01-01

In a recent paper we have shown that DNA haplotypes of multiply heterozygous individuals can be resolved directly by polymerase-chain-reaction (PCR) amplification of a single molecule of genomic template. Our method (the single-molecule-dilution [SMD] method) relies on the stochastic separation of maternal and paternal alleles at high dilution. The stochasticity of separation and the potential for DNA shearing (which could separate the loci of interest) are two factors that can compromise the results of the experiment. This paper explores the consequences of these two factors and shows that the SMD method can be expected to work very reliably even in the presence of a moderate amount of DNA shearing. PMID:2339707

Chemical Disposal for a High School Chemistry Laboratory

ERIC Educational Resources Information Center

Mento, Mary Ann

1973-01-01

A method of disposal is suggested that is superior to the dilution method. The proceudres are based on the conversion of the wastes to less toxic or harmless forms or neutralization, so that they won't be a shock to the sewage system, and then excess dilution. (Author/DF)

The Effect of Albumin and Platelet-Poor Plasma Supplemented Botulinum A Toxin on Bioavaliability: An Experimental Rabbit Model.

PubMed

Sibar, Serhat; Findikcioglu, Kemal; Zinnuroglu, Murat; Cenetoglu, Seyhan

2017-04-01

Today, botulinum toxin is commonly used for cosmetic purposes throughout the world. Despite various agents reducing the efficiency of toxin are well defined, the studies related to increasing the bioavailability are limited. The purpose of our study is to assess the effect of the preparation of toxin by diluting with platelet-poor plasma (PPP) and/or albumin instead of standard dilution (saline) on bioavailability in cosmetic-purpose botulinum toxin applications.In the study, 24 New Zealand rabbits were used. Right anterior auricular muscle was preferred for toxin injections. Subjects were divided in 4 groups and in every group; botulinum A toxin (BTxA) that was prepared by different dilution methods was injected. 2.5 U saline-diluted BTxA was injected to the subjects in group 1, 2.5 U ready-to-use rabbit albumin-diluted BTxA was injected to group 2 and 2.5 U autologous PPP-diluted BTxA was injected to group 3 and pure saline was injected to group 4.Before the injection (0th week) and in the second, sixth, and 12th weeks after the injection, visual and electroneuromyographic evaluations of the ears of the subjects were performed.In the second week, median amplitude levels in group 2 were significantly found lower than other groups.In the sixth week, median amplitude levels in group 1 were significantly found lower than other groups.In 12th week, no significant difference was found among all the groups in terms of median amplitude levels.Visual findings were also correlated with electroneuromyographic findings.It was observed that the dilution of BTxA with albumin had caused a stronger paralysis when compared to dilution with saline or PPP at the beginning (second week); however, in the following weeks (sixth week), it was seen that dilution with saline had maintained paralysis better when compared with other dilution methods.In cosmetic BTxA applications, dilution of the toxin with albumin or PPP instead of standard dilution has no positive effect on bioavailability and such modifications regarding this kind of dilution are found unsuitable. Further studies are needed to directly relate the results with clinical applications.

Randomly diluted eg orbital-ordered systems.

PubMed

Tanaka, T; Matsumoto, M; Ishihara, S

2005-12-31

Dilution effects on the long-range ordered state of the doubly degenerate e(g) orbital are investigated. Quenched impurities without the orbital degree of freedom are introduced in the orbital model where the long-range order is realized by the order-from-disorder mechanism. It is shown by Monte Carlo simulations and the cluster-expansion method that a decrease in the orbital-ordering temperature by dilution is substantially larger than that in the randomly diluted spin models. Tilting of orbital pseudospins around impurities is the essence of this dilution effect. The present theory provides a new viewpoint for the recent resonant x-ray scattering experiments in KCu(1-x)Zn(x)F(3).

Dilute acid/metal salt hydrolysis of lignocellulosics

DOEpatents

Nguyen, Quang A.; Tucker, Melvin P.

2002-01-01

A modified dilute acid method of hydrolyzing the cellulose and hemicellulose in lignocellulosic material under conditions to obtain higher overall fermentable sugar yields than is obtainable using dilute acid alone, comprising: impregnating a lignocellulosic feedstock with a mixture of an amount of aqueous solution of a dilute acid catalyst and a metal salt catalyst sufficient to provide higher overall fermentable sugar yields than is obtainable when hydrolyzing with dilute acid alone; loading the impregnated lignocellulosic feedstock into a reactor and heating for a sufficient period of time to hydrolyze substantially all of the hemicellulose and greater than 45% of the cellulose to water soluble sugars; and recovering the water soluble sugars.

Entrainment vs. Dilution in Tropical Deep Convection

NASA Astrophysics Data System (ADS)

Hannah, W.

2017-12-01

The distinction between entrainment and dilution is investigated with cloud resolving simulations of deep convection in a tropical environment. A method for estimating the rate of dilution by entrainment and detrainment is calculated for a series of bubble simulations with a range of initial radii. Entrainment generally corresponds to dilution of convection, but the two quantities are not well correlated. Core dilution by entrainment is significantly reduced by the presence of a shell of moist air around the core. Entrainment contributes significantly to the total net dilution, but detrainment and the various source/sink terms play large roles depending on the variable in question. Detrainment has a concentrating effect on average that balances out the dilution by entrainment. The experiments are also used to examine whether entrainment or dilution scale with cloud radius. The results support a weak negative relationship for dilution, but not for entrainment. The sensitivity to resolution is briefly discussed. A toy Lagrangian thermal model is used to demonstrate the importance of the cloud shell as a thermodynamic buffer to reduce the dilution of the core by entrainment. The results suggest that explicit cloud heterogeneity may be a useful consideration for future convective parameterization development.

Dilute-and-shoot coupled to nanoflow liquid chromatography high resolution mass spectrometry for the determination of drugs of abuse and sport drugs in human urine.

PubMed

Alcántara-Durán, Jaime; Moreno-González, David; Beneito-Cambra, Miriam; García-Reyes, Juan F

2018-05-15

In this work, a sensitive nanoflow liquid chromatography high-resolution mass spectrometry screening method has been developed for the determination of multiclass drugs of abuse and sport drugs in human urine. 81 drugs belonging to different multiclass pharmaceuticals were targeted. The method is based on the use of a nanoLC column (75 µm × 150 mm, 3 µm particle size and 100 Å pore) with the nanospray emitter tip integrated so that dead volumes are significantly minimized. Data acquisition method included both full-scan and all ion fragmentation experiments using an Orbitrap analyser (Q-Exactive) operated in the positive ionization mode. To increase laboratory throughput, a dilute-and-shoot methodology has been tested and proposed, based solely on direct urine dilution without further sample workup. Matrix effects were evaluated, showing a negligible effect for all studied compounds when a dilution 1:50 was implemented. Despite this high-dilution factor, limits of quantification were still satisfactory, with values below 5 µg L -1 in most cases, being lower than their minimum required performance limits correspond established by the World Anti-Doping Agency. Therefore, the use of the dilute-and-shoot method with the enhanced sensitivity provided by nanoflow LC setup could be useful tool for the determination of studied compounds in drug testing, thus increasing laboratory performance, because a minimum sample treatment steps are required. Copyright © 2018 Elsevier B.V. All rights reserved.

Determination of buildup and dilution of wastewater effluent in shellfish growing waters through a modified application of super-position.

PubMed

Goblick, Gregory N; Ao, Yaping; Anbarchian, Julie M; Calci, Kevin R

2017-02-15

Since 1925, dilution analysis has been used to minimize pathogenic impacts to bivalve molluscan shellfish growing areas from treated wastewater effluent in the National Shellfish Sanitation Program (NSSP). For over twenty five years, the U.S. Food and Drug Administration (FDA) has recommended a minimum of 1000:1 dilution of effluent within prohibited closure zones established around wastewater treatment plant (WWTP) discharges. During May 2010, using recent technologies, a hydrographic dye study was conducted in conjunction with a pathogen bioaccumulation study in shellfish adjacent to a WWTP discharge in Yarmouth, ME. For the first time an improved method of the super-position principle was used to determine the buildup of dye tagged sewage effluent and steady state dilution in tidal waters. Results of the improved method of dilution analysis illustrate an economical, reliable and more accurate and manageable approach for estimating the buildup and steady state pollutant conditions in coastal and estuarine waters. Published by Elsevier Ltd.

Entrainment versus Dilution in Tropical Deep Convection

DOE PAGES

Hannah, Walter M.

2017-11-01

In this paper, the distinction between entrainment and dilution is investigated with cloud-resolving simulations of deep convection in a tropical environment. A method for estimating the rate of dilution by entrainment and detrainment is presented and calculated for a series of bubble simulations with a range of initial radii. Entrainment generally corresponds to dilution of convection, but the two quantities are not well correlated. Core dilution by entrainment is significantly reduced by the presence of a shell of moist air around the core. Dilution by entrainment also increases with increasing updraft velocity but only for sufficiently strong updrafts. Entrainment contributesmore » significantly to the total net dilution, but detrainment and the various source/sink terms play large roles depending on the variable in question. Detrainment has a concentrating effect on average that balances out the dilution by entrainment. The experiments are also used to examine whether entrainment or dilution scale with cloud radius. The results support a weak negative relationship for dilution but not for entrainment. The sensitivity to resolution is briefly discussed. A toy Lagrangian thermal model is used to demonstrate the importance of the cloud shell as a thermodynamic buffer to reduce the dilution of the core by entrainment. Finally, the results suggest that explicit cloud heterogeneity may be a useful consideration for future convective parameterization development.« less

A helium-3/helium-4 dilution cryocooler for operation in zero gravity

NASA Technical Reports Server (NTRS)

Hendricks, John B.

1988-01-01

This research effort covered the development of He-3/He-4 dilution cryocooler cycles for use in zero gravity. The dilution cryocooler is currently the method of choice for producing temperatures below 0.3 Kelvin in the laboratory. However, the current dilution cryocooler depends on gravity for their operation, so some modification is required for zero gravity operation. In this effort, we have demonstrated, by analysis, that the zero gravity dilution cryocooler is feasible. We have developed a cycle that uses He-3 circulation, and an alternate cycle that uses superfluid He-4 circulation. The key elements of both cycles were demonstrated experimentally. The development of a true 'zero-gravity' dilution cryocooler is now possible, and should be undertaken in a follow-on effort.

Performance of Two Quantitative PCR Methods for Microbial Source Tracking of Human Sewage and Implications for Microbial Risk Assessment in Recreational Waters

PubMed Central

Staley, Christopher; Gordon, Katrina V.; Schoen, Mary E.

2012-01-01

Before new, rapid quantitative PCR (qPCR) methods for assessment of recreational water quality and microbial source tracking (MST) can be useful in a regulatory context, an understanding of the ability of the method to detect a DNA target (marker) when the contaminant source has been diluted in environmental waters is needed. This study determined the limits of detection and quantification of the human-associated Bacteroides sp. (HF183) and human polyomavirus (HPyV) qPCR methods for sewage diluted in buffer and in five ambient, Florida water types (estuarine, marine, tannic, lake, and river). HF183 was quantifiable in sewage diluted up to 10−6 in 500-ml ambient-water samples, but HPyVs were not quantifiable in dilutions of >10−4. Specificity, which was assessed using fecal composites from dogs, birds, and cattle, was 100% for HPyVs and 81% for HF183. Quantitative microbial risk assessment (QMRA) estimated the possible norovirus levels in sewage and the human health risk at various sewage dilutions. When juxtaposed with the MST marker detection limits, the QMRA analysis revealed that HF183 was detectable when the modeled risk of gastrointestinal (GI) illness was at or below the benchmark of 10 illnesses per 1,000 exposures, but the HPyV method was generally not sensitive enough to detect potential health risks at the 0.01 threshold for frequency of illness. The tradeoff between sensitivity and specificity in the MST methods indicates that HF183 data should be interpreted judiciously, preferably in conjunction with a more host-specific marker, and that better methods of concentrating HPyVs from environmental waters are needed if this method is to be useful in a watershed management or monitoring context. PMID:22885746

RM Methods for Multiple Fare Structure Environments

DTIC Science & Technology

2008-06-01

110 9 LIST OF FIGURES 1 INTRODUCTION Figure 1-1: Revenue Losses due to Overprotection and Dilution... overprotection or dilution. If an airline’s objective is to focus on high-yield business travelers and charge only a single high fare for the...Lost Revenue from Customer Surplus Captured Revenue Rejected Passengers Overprotection Dilution Figure 1-1: Revenue Losses due to Overprotection

Quantifying the dilution effect for models in ecological epidemiology.

PubMed

Roberts, M G; Heesterbeek, J A P

2018-03-01

The dilution effect , where an increase in biodiversity results in a reduction in the prevalence of an infectious disease, has been the subject of speculation and controversy. Conversely, an amplification effect occurs when increased biodiversity is related to an increase in prevalence. We explore the conditions under which these effects arise, using multi species compartmental models that integrate ecological and epidemiological interactions. We introduce three potential metrics for quantifying dilution and amplification, one based on infection prevalence in a focal host species, one based on the size of the infected subpopulation of that species and one based on the basic reproduction number. We introduce our approach in the simplest epidemiological setting with two species, and show that the existence and strength of a dilution effect is influenced strongly by the choices made to describe the system and the metric used to gauge the effect. We show that our method can be generalized to any number of species and to more complicated ecological and epidemiological dynamics. Our method allows a rigorous analysis of ecological systems where dilution effects have been postulated, and contributes to future progress in understanding the phenomenon of dilution in the context of infectious disease dynamics and infection risk. © 2018 The Author(s).

Electrodeionization method

DOEpatents

Lin, YuPo J.; Hestekin, Jamie; Arora, Michelle; St. Martin, Edward J.

2004-09-28

An electrodeionization method for continuously producing and or separating and/or concentrating ionizable organics present in dilute concentrations in an ionic solution while controlling the pH to within one to one-half pH unit method for continuously producing and or separating and/or concentrating ionizable organics present in dilute concentrations in an ionic solution while controlling the pH to within one to one-half pH unit.

In Situ Local Fracture Flow Measurement by the Double Packer Dilution Test

NASA Astrophysics Data System (ADS)

Englert, A.; Le Borgne, T.; Bour, O.; Klepikova, M.; Lavenant, N.

2011-12-01

For prediction of flow and transport in fractured media, prior estimation of the fracture network is essential, but challenging. Recent developments in hydraulic tomography have shown promising results for understanding connectivities between boreholes. However, as the hydraulic tomographic survey is typically based on the propagation of head only, it becomes a strongly non unique problem. To reduce the non uniqueness of tomographic surveys point conditioning has been found beneficial. Just as well, measurement of local flow in a fracture can serve as point conditioning for hydraulic and tracer tomographic surveys. Nevertheless, only few measurements of local fracture flow have been performed since this type of measurements implies several important technical issues. Dilution test in a packed off interval is a possible method for measuring fracture flow (e.g. Drost et al. 1968, Novakowski et al., 2005). However, a key issue for estimating flow with dilution tests is to ensure a full mixing of the tracer in the packed interval. This is typically done by including a mixing system within the packer. The design of such system can be challenging for deep wells and small diameters. Here, we propose a method where mixing is ensured by a recirculation loop including a surface tank. This method is adapted from the design proposed by Brouyere et al. (2008), who measured dilution in open wells. Dilution is quantified by measuring the concentration in the surface barrel as function of time. Together with the measurement of the circulating flow and the water filled volume in the surface barrel, the measured tracer dilution allows for calculation of the fracture flow. Since the method can be applied using a classical double packer system, it may provide a broader application of local flow measurements in heterogeneous media. We tested the approach on the Ploemeur fractured crystalline rock site. A one meter interval at depth 80 m with a single flowing fracture was isolated with a double packer dilution system. We performed a pumping test in the adjacent well. Different flow rates were estimated from the dilution curves for the different pumping rates in the adjacent well, showing a linear response. The obtained fracture flow rates provide important information on the flow geometry and connectivity between the two wells. Future joint interpretation of flow measurements, hydraulic head and tracer test data is expected to provide detailed insights in the flow and transport processes at the Ploemeur site. Drost, W., Klotz, D., Koch, A., Moser, H., Neumaier, F., Rauert, W.: Point dilution methods of investigating ground water flow by means of radioisotopes, Water. Resour. Res., 4(1), 1968. Novakowski, K., Bickerton, G., Lapcevic, P., Voralek, J., Ross, N.: Measurements of groundwater velocity in discrete rock fractures: Jour. Cont. Hydr., 82(1-2), 2006. Brouyere, S., Batlle-Aguilar, J., Goderniaux, P., Dassargues, A.: A new tracer technique for monitoring groundwater fluxes: The Finite Volume Point Dilution Method, Jour. Cont. Hydr., 95(3-4), 121-140, 2008.

Comparative In Vitro Efficacy of Doripenem and Imipenem Against Multi-Drug Resistant Pseudomonas aeruginosa.

PubMed

Wali, Nadia; Mirza, Irfan Ali

2016-04-01

To compare the in vitro efficacy of doripenem and imipenem against multi-drug resistant (MDR) Pseudomonas aeruginosa from various clinical specimens. Descriptive cross-sectional study. Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from November 2012 to November 2013. MDR Pseudomonas aeruginosa isolates from various clinical samples were included in the study. Susceptibility of Pseudomonas aeruginosa against doripenem and imipenem was performed by E-test strip and agar dilution methods. The results were interpreted as recommended by Clinical Laboratory Standard Institute (CLSI) guidelines. The maximum number of Pseudomonas aeruginosa were isolated from pure pus and pus swabs. In vitro efficacy of doripenem was found to be more effective as compared to imipenem against MDR Pseudomonas aeruginosa with both E-test strip and agar dilution methods. Overall, p-values of 0.014 and 0.037 were observed when susceptibility patterns of doripenem and imipenem were evaluated with E-test strip and agar dilution methods. In vitro efficacy of doripenem was found to be better against MDR Pseudomonas aeruginosaas compared to imipenem when tested by both E-test and agar dilution methods.

Single polymer dynamics in semi-dilute unentangled and entangled solutions: from molecular conformation to normal stress

NASA Astrophysics Data System (ADS)

Schroeder, Charles

Semi-dilute polymer solutions are encountered in a wide array of applications such as advanced 3D printing technologies. Semi-dilute solutions are characterized by large fluctuations in concentration, such that hydrodynamic interactions, excluded volume interactions, and transient chain entanglements may be important, which greatly complicates analytical modeling and theoretical treatment. Despite recent progress, we still lack a complete molecular-level understanding of polymer dynamics in these systems. In this talk, I will discuss three recent projects in my group to study semi-dilute solutions that focus on single molecule studies of linear and ring polymers and a new method to measure normal stresses in microfluidic devices based on the Stokes trap. In the first effort, we use single polymer techniques to investigate the dynamics of semi-dilute unentangled and semi-dilute entangled DNA solutions in extensional flow, including polymer relaxation from high stretch, transient stretching dynamics in step-strain experiments, and steady-state stretching in flow. In the semi-dilute unentangled regime, our results show a power-law scaling of the longest polymer relaxation time that is consistent with scaling arguments based on the double cross-over regime. Upon increasing concentration, we observe a transition region in dynamics to the entangled regime. We also studied the transient and steady-state stretching dynamics in extensional flow using the Stokes trap, and our results show a decrease in transient polymer stretch and a milder coil-to-stretch transition for semi-dilute polymer solutions compared to dilute solutions, which is interpreted in the context of a critical Weissenberg number Wi at the coil-to-stretch transition. Interestingly, we observe a unique set of polymer conformations in semi-dilute unentangled solutions that are highly suggestive of transient topological entanglements in solutions that are nominally unentangled at equilibrium. Taken together, these results suggest that the transient stretching pathways in semi-dilute solution extensional flows are qualitatively different than for both dilute solutions and for semi-dilute solutions in shear flow. In a second effort, we studied the dynamics of ring polymers in background solutions of semi-dilute linear polymers. Interestingly, we observe strikingly large fluctuations in steady-state polymer extension for ring polymers in flow, which occurs due to the interplay between polymer topology and concentration leading to chain `threading' in flow. In a third effort, we developed a new microfluidic method to measure normal stress and extensional viscosity that can be loosely described as passive yet non-linear microrheology. In particular, we incorporated 3-D particle imaging velocimetry (PIV) with the Stokes trap to study extensional flow-induced particle migration in semi-dilute polymer solutions. Experimental results are analyzed using the framework of a second-order-fluid model, which allows for measurement of normal stress and extensional viscosity in semi-dilute polymer solutions, all of which is a first-of-its-kind demonstration. Microfluidic measurements of extensional viscosity are directly compared to the dripping-onto-substrate or DOS method, and good agreement is generally observed. Overall, our work aims to provide a molecular-level understanding of the role of polymer topology and concentration on bulk rheological properties by using single polymer techniques.

Use of a novel radiometric method to assess the inhibitory effect of donepezil on acetylcholinesterase activity in minimally diluted tissue samples

PubMed Central

Kikuchi, Tatsuya; Okamura, Toshimitsu; Arai, Takuya; Obata, Takayuki; Fukushi, Kiyoshi; Irie, Toshiaki; Shiraishi, Tetsuya

2010-01-01

Background and purpose: Cholinesterase inhibitors have been widely used for the treatment of patients with dementia. Monitoring of the cholinesterase activity in the blood is used as an indicator of the effect of the cholinesterase inhibitors in the brain. The selective measurement of cholinesterase with low tissue dilution is preferred for accurate monitoring; however, the methods have not been established. Here, we investigated the effect of tissue dilution on the action of cholinesterase inhibitors using a novel radiometric method with selective substrates, N-[14C]methylpiperidin-4-yl acetate ([14C]MP4A) and (R)-N-[14C]methylpiperidin-3-yl butyrate ([14C]MP3B_R), for AChE and butyrylcholinesterase (BChE) respectively. Experimental approach: We investigated the kinetics of hydrolysis of [14C]-MP4A and [14C]-MP3B_R by cholinesterases, and evaluated the selectivity of [14C]MP4A and [14C]MP3B_R for human AChE and BChE, respectively, compared with traditional substrates. Then, IC50 values of cholinesterase inhibitors in minimally diluted and highly diluted tissues were measured with [14C]MP4A and [14C]MP3B_R. Key results: AChE and BChE activities were selectively measured as the first-order hydrolysis rates of [14C]-MP4A and [14C]MP3B_R respectively. The AChE selectivity of [14C]MP4A was an order of magnitude higher than traditional substrates used for the AChE assay. The IC50 values of specific AChE and BChE inhibitors, donepezil and ethopropazine, in 1.2-fold diluted human whole blood were much higher than those in 120-fold diluted blood. In addition, the IC50 values of donepezil in monkey brain were dramatically decreased as the tissue was diluted. Conclusions and implications: This method would effectively monitor the activity of cholinesterase inhibitors used for therapeutics, pesticides and chemical warfare agents. PMID:20401964

Sonic wave separation of invertase from a dilute solution to generated droplets.

PubMed

Tanner, R D; Ko, S; Loha, V; Prokop, A

2000-01-01

It has previously been shown that a droplet fractionation process, simulated by shaking a separatory funnel containing a dilute protein solution, can generate droplets richer in protein than present in the original dilute solution. In this article, we describe an alternative method that can increase the amount of protein transferred to the droplets. The new method uses ultrasonic waves, enhanced by a bubble gas stream to create the droplets. The amount of protein in these droplets increases by about 50%. In this method, the top layer of the dilute protein solution (of the solution-air interface) becomes enriched in protein when air is bubbled into the solution. This concentrating procedure is called bubble fractionation. Once the protein has passed through the initial buildup, this enriched protein layer is transferred into droplets with the aid of a vacuum above the solution at the same time that ultrasonic waves are introduced. The droplets are then carried over to a condenser and coalesced. We found that this new method provides an easier way to remove the protein-enriched top layer of the dilute solution and generates more droplets within a shorter period than the separatory funnel droplet generation method. The added air creates the bubbles and carries the droplets, and the vacuum helps remove the effluent airstream from the condenser. The maximum partition coefficient, the ratio of the protein concentration in the droplets to that in the residual solution (approx 8.5), occurred at pH 5.0.

ESTIMATION OF THE NUMBER OF INFECTIOUS BACTERIAL OR VIRAL PARTICLES BY THE DILUTION METHOD

PubMed Central

Seligman, Stephen J.; Mickey, M. Ray

1964-01-01

Seligman, Stephen J. (University of California, Los Angeles), and M. Ray Mickey. Estimation of the number of infectious bacterial or viral particles by the dilution method. J. Bacteriol. 88:31–36. 1964.—For viral or bacterial systems in which discrete foci of infection are not obtainable, it is possible to obtain an estimate of the number of infectious particles by use of the quantal response if the assay system is such that one infectious particle can elicit the response. Unfortunately, the maximum likelihood estimate is difficult to calculate, but, by the use of a modification of Haldane's approximation, it is possible to construct a table which facilitates calculation of both the average number of infectious particles and its relative error. Additional advantages of the method are that the number of test units per dilution can be varied, the dilutions need not bear any fixed relation to each other, and the one-particle hypothesis can be readily tested. PMID:14197902

Isotope dilution ICP-MS with laser-assisted sample introduction for direct determination of sulfur in petroleum products.

PubMed

Boulyga, Sergei F; Heilmann, Jens; Heumann, Klaus G

2005-08-01

Inductively coupled plasma isotope dilution mass spectrometry (ICP-IDMS) with direct laser-assisted introduction of isotope-diluted samples into the plasma, using a laser ablation system with high ablation rates, was developed for accurate sulfur determinations in different petroleum products such as 'sulfur-free' premium gasoline, diesel fuel, and heating oil. Two certified gas oil reference materials were analyzed for method validation. Two different 34S-enriched spike compounds, namely, elementary sulfur dissolved in xylene and dibenzothiophene in hexane, were synthesized and tested for their usefulness in this isotope dilution technique. The isotope-diluted sample was adsorbed on a filter-paper-like material, which was fixed in a special holder for irradiation by the laser beam. Under these conditions no time-dependent spike/analyte fractionation was only observed for the dibenzothiophene spike during the laser ablation process, which means that the measured 34S/32S isotope ratio of the isotope-diluted sample remained constant-a necessary precondition for accurate results with the isotope dilution technique. A comparison of LA-ICP-IDMS results with the certified values of the gas oil reference materials and with results obtained from ICP-IDMS analyses with wet sample digestion demonstrated the accuracy of the new LA-ICP-IDMS method in the concentration range of 9.2 microg g(-1) ('sulfur-free' premium gasoline) to 10.4 mg g(-1) (gas oil reference material BCR 107). The detection limit for sulfur by LA-ICP-IDMS is 0.04 microg g(-1) and the analysis time is only about 10 min, which therefore also qualifies this method for accurate determinations of low sulfur contents in petroleum products on a routine level.

[Polymerase chain reaction (PCR) for the identification of toxigenic Vibrio cholerae O1 in oysters].

PubMed

Rodríguez-Angeles, M G; Giono-Cerezo, S; Moreno-Escobar, A; Valdespino-Gómez, J L

1994-01-01

PCR was made with ctx2 (CGG GCA GAT TCT AGA CCT CCT G) y ctx3 (CGA TGA TCT TGG AGC ATT CCC AC) primers for subunit A of cholera toxin, 30 cycles of temperature on samples of 50 g of oysters added in 450 ml of peptone alcaline water that were inoculated with 15 x 10(6), 0.75 x 10(6) and 0.15 x 10(6) CFU/ml of toxigenic 6707 V. cholerae O1 reference strain. The samples were tested by three microbiological methods: INDRE's method uses 1 x 10(-1) dilution of sample, two fold pass to peptone alcaline water pH 9 incubated 18 h and 6 h at 37 degrees C, the Food and Drugs Administration (FDA) method uses 10(-1) to 10(-6) dilutions of sample, 6 h incubation and reincubation for 18 h at 37 and 42 degrees C and the Mexican laboratories (LMD) with 10(-4) to 10(-3) dilutions, the samples were incubated for 6 h and then reincubated for 18 h at two temperatures 37 and 42 degrees C. The PCR by INDRE's method was positive with 3 x 10(2) CFU/ml/g oyster. In the FDA's method the PCR detected DNA in 10(-4) dilution with 3 x 10(1) CFU/ml/g oyster and in LMD's method the PCR was positive in 10(-3) with 3 CFU/ml/g oyster. The results of the PCR were obtained between 5-6 h, and later V. cholerae O1 was isolated by three microbiological methods. The PCR reproducibility was better on DNA sample diluted 1:4 and 10 microliters of sample increased from 1:1000 to 1:10000 the sensitivity of PCR.

An accurate algorithm for the detection of DNA fragments from dilution pool sequencing experiments.

PubMed

Bansal, Vikas

2018-01-01

The short read lengths of current high-throughput sequencing technologies limit the ability to recover long-range haplotype information. Dilution pool methods for preparing DNA sequencing libraries from high molecular weight DNA fragments enable the recovery of long DNA fragments from short sequence reads. These approaches require computational methods for identifying the DNA fragments using aligned sequence reads and assembling the fragments into long haplotypes. Although a number of computational methods have been developed for haplotype assembly, the problem of identifying DNA fragments from dilution pool sequence data has not received much attention. We formulate the problem of detecting DNA fragments from dilution pool sequencing experiments as a genome segmentation problem and develop an algorithm that uses dynamic programming to optimize a likelihood function derived from a generative model for the sequence reads. This algorithm uses an iterative approach to automatically infer the mean background read depth and the number of fragments in each pool. Using simulated data, we demonstrate that our method, FragmentCut, has 25-30% greater sensitivity compared with an HMM based method for fragment detection and can also detect overlapping fragments. On a whole-genome human fosmid pool dataset, the haplotypes assembled using the fragments identified by FragmentCut had greater N50 length, 16.2% lower switch error rate and 35.8% lower mismatch error rate compared with two existing methods. We further demonstrate the greater accuracy of our method using two additional dilution pool datasets. FragmentCut is available from https://bansal-lab.github.io/software/FragmentCut. vibansal@ucsd.edu. Supplementary data are available at Bioinformatics online. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Apparatus and method for maintaining multi-component sample gas constituents in vapor phase during sample extraction and cooling

DOEpatents

Felix, Larry Gordon; Farthing, William Earl; Irvin, James Hodges; Snyder, Todd Robert

2010-05-11

A dilution apparatus for diluting a gas sample. The apparatus includes a sample gas conduit having a sample gas inlet end and a diluted sample gas outlet end, and a sample gas flow restricting orifice disposed proximate the sample gas inlet end connected with the sample gas conduit and providing fluid communication between the exterior and the interior of the sample gas conduit. A diluted sample gas conduit is provided within the sample gas conduit having a mixing end with a mixing space inlet opening disposed proximate the sample gas inlet end, thereby forming an annular space between the sample gas conduit and the diluted sample gas conduit. The mixing end of the diluted sample gas conduit is disposed at a distance from the sample gas flow restricting orifice. A dilution gas source connected with the sample gas inlet end of the sample gas conduit is provided for introducing a dilution gas into the annular space, and a filter is provided for filtering the sample gas. The apparatus is particularly suited for diluting heated sample gases containing one or more condensable components.

A comparative evaluation of six principal IgY antibody extraction methods.

PubMed

Ren, Hao; Yang, Wenjing; Thirumalai, Diraviyam; Zhang, Xiaoying; Schade, Rüdiger

2016-03-01

Egg yolk has been considered a promising source of antibodies. Our study was designed to compare six principal IgY extraction methods (water dilution, polyethylene glycol [PEG] precipitation, caprylic acid extraction, chloroform extraction, phenol extraction, and carrageenan extraction), and to assess their relative extraction efficiencies and the purity of the resulting antibodies. The results showed that the organic solvents (chloroform or phenol) minimised the lipid ratio in the egg yolk. The water dilution, PEG precipitation and caprylic acid extraction methods resulted in high yields, and antibodies purified with PEG and carrageenan exhibited high purity. Our results indicate that phenol extraction would be more suitable for preparing high concentrations of IgY for non-therapeutic usage, while the water dilution and carrageenan extraction methods would be more appropriate for use in the preparation of IgY for oral administration. 2016 FRAME.

DETERMINATION OF THE MAXIMUM INHIBITORY DILUTION OF CETYLPYRIDINIUM CHLORIDE-BASED MOUTHWASHES AGAINST STAPHYLOCOCCUS AUREUS: AN IN VITRO STUDY

PubMed Central

Watanabe, Evandro; Tanomaru, Juliane Maria Guerreiro; Nascimento, Andresa Piacezzi; Matoba, Fumio; Tanomaru, Mario; Ito, Izabel Yoko

2008-01-01

The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)-based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Juá+Romã+Propolis (Natural Honey®) and CPC (Cepacol®), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/ 655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37°C for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Juá+Romã+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol® showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol® was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect. PMID:19089260

Measurements of left-to-right intracardiac shunting in adults: oximetric versus indicator dilution techniques.

PubMed

Hillis, L D; Winniford, M D; Jackson, J A; Firth, B G

1985-01-01

Of the various techniques that are available to assess the presence and magnitude of intracardiac shunting, oximetry and indicator dilution are used most frequently. This study was performed to compare these methods in adult patients with pure left-to-right intracardiac shunting. In 27 patients [12 men, 15 women, aged 32 +/- 12 (mean +/- SD) years], shunt magnitude was assessed in close temporal proximity by both techniques. The oximetric percentage left-to-right shunt averaged 55 +/- 16%, whereas the indocyanine green dye percentage shunt was significantly less (36 +/- 14%, p less than 0.001). The oximetric percentage shunt exceeded the indocyanine green dye percentage shunt in 26 and did so by greater than 20% in 23 patients. Thus, there is a substantial difference in shunt magnitude between the oximetric and the indicator dilution techniques. In infants, indicator dilution yields results that are larger than those obtained with the oximetric method. Conversely, in adults, the indicator dilution technique gives results that are consistently smaller than those obtained with oximetry. As a result, the decision regarding therapy of an intracardiac shunt should be made with this in mind.

High accuracy method for the application of isotope dilution to gas chromatography/mass spectrometric analysis of gases.

PubMed

Milton, Martin J T; Wang, Jian

2003-01-01

A new isotope dilution mass spectrometry (IDMS) method for high-accuracy quantitative analysis of gases has been developed and validated by the analysis of standard mixtures of carbon dioxide in nitrogen. The method does not require certified isotopic reference materials and does not require direct measurements of the highly enriched spike. The relative uncertainty of the method is shown to be 0.2%. Reproduced with the permission of Her Majesty's Stationery Office. Copyright Crown copyright 2003.

Determination of copper by isotopic dilution.

PubMed

Faquim, E S; Munita, C S

1994-01-01

A rapid and selective method was used for the determination of copper by isotopic dilution employing substoichiometric extraction with dithizone in carbon tetrachloride. The appropriate pH range for the substoichiometric extraction was 2-7. In the analysis, even a large excess of elements forming extractable complexes with dithizone does not interfere. The accuracy and precision of the method were evaluated. The method has been applied to analysis of reference materials, wheat flour, wine, and beer.

In Vitro Activities of Gemifloxacin versus Five Quinolones and Two Macrolides against 271 Spanish Isolates of Legionella pneumophila: Influence of Charcoal on Susceptibility Test Results

PubMed Central

García, M. T.; Pelaz, C.; Giménez, M. J.; Aguilar, L.

2000-01-01

The MICs at which 90% of isolates are inhibited for gemifloxacin, trovafloxacin, and grepafloxacin were low (≤0.01 μg/ml) for 271 Legionella isolates when they were determined by the broth microdilution method but increased (≥6 dilutions) when they were determined by the agar dilution method. This was due to the charcoal in the agar dilution medium, as shown by the progressive decrease in the MICs when the charcoal concentrations decreased. As free drug is the active fraction, charcoal binding should be considered. PMID:10898695

Apoplastic water fraction and rehydration techniques introduce significant errors in measurements of relative water content and osmotic potential in plant leaves.

PubMed

Arndt, Stefan K; Irawan, Andi; Sanders, Gregor J

2015-12-01

Relative water content (RWC) and the osmotic potential (π) of plant leaves are important plant traits that can be used to assess drought tolerance or adaptation of plants. We estimated the magnitude of errors that are introduced by dilution of π from apoplastic water in osmometry methods and the errors that occur during rehydration of leaves for RWC and π in 14 different plant species from trees, grasses and herbs. Our data indicate that rehydration technique and length of rehydration can introduce significant errors in both RWC and π. Leaves from all species were fully turgid after 1-3 h of rehydration and increasing the rehydration time resulted in a significant underprediction of RWC. Standing rehydration via the petiole introduced the least errors while rehydration via floating disks and submerging leaves for rehydration led to a greater underprediction of RWC. The same effect was also observed for π. The π values following standing rehydration could be corrected by applying a dilution factor from apoplastic water dilution using an osmometric method but not by using apoplastic water fraction (AWF) from pressure volume (PV) curves. The apoplastic water dilution error was between 5 and 18%, while the two other rehydration methods introduced much greater errors. We recommend the use of the standing rehydration method because (1) the correct rehydration time can be evaluated by measuring water potential, (2) overhydration effects were smallest, and (3) π can be accurately corrected by using osmometric methods to estimate apoplastic water dilution. © 2015 Scandinavian Plant Physiology Society.

A method for measuring particle number emissions from vehicles driving on the road.

PubMed

Shi, J P; Harrison, R M; Evans, D E; Alam, A; Barnes, C; Carter, G

2002-01-01

Earlier research has demonstrated that the conditions of dilution of engine exhaust gases profoundly influence the size distribution and total number of particles emitted. Since real world dilution conditions are variable and therefore difficult to simulate, this research has sought to develop and validate a method for measuring particle number emissions from vehicles driving past on a road. This has been achieved successfully using carbon dioxide as a tracer of exhaust gas dilution. By subsequent adjustment of data to a constant dilution factor, it is possible to compare emissions from different vehicles using different technologies and fuels based upon real world emission data. Whilst further optimisation of the technique, especially in terms of matching the instrument response times is desirable, the measurements offer useful insights into emissions from gasoline and diesel vehicles, and the substantial proportion of particles emitted in the 3-7 nanometre size range.

Chemical footprint method for improved communication of freshwater ecotoxicity impacts in the context of ecological limits.

PubMed

Bjørn, Anders; Diamond, Miriam; Birkved, Morten; Hauschild, Michael Zwicky

2014-11-18

The ecological footprint method has been successful in communicating environmental impacts of anthropogenic activities in the context of ecological limits. We introduce a chemical footprint method that expresses ecotoxicity impacts from anthropogenic chemical emissions as the dilution needed to avoid freshwater ecosystem damage. The indicator is based on USEtox characterization factors with a modified toxicity reference point. Chemical footprint results can be compared to the actual dilution capacity within the geographic vicinity receiving the emissions to estimate whether its ecological limit has been exceeded and hence whether emissions can be expected to be environmentally sustainable. The footprint method was illustrated using two case studies. The first was all inventoried emissions from European countries and selected metropolitan areas in 2004, which indicated that the dilution capacity was likely exceeded for most European countries and all landlocked metropolitan areas. The second case study indicated that peak application of pesticides alone was likely to exceed Denmark's freshwater dilution capacity in 1999-2011. The uncertainty assessment showed that better spatially differentiated fate factors would be useful and pointed out other major sources of uncertainty and some opportunities to reduce these.

Comparative activity of several beta-lactam antibiotics against anaerobes determined by two methods.

PubMed

Zabransky, R J; Birk, R J

1987-01-01

The susceptibility of 120 strains of several species of anaerobes to a number of second and third generation beta-lactam antibiotics was determined by the National Committee for Clinical Laboratory Standards reference agar dilution and microdilution methods. The antibiotics tested were cefoperazone, cefotaxime, cefotetan, ceftizoxime, cefoxitin, and imipenem. The MIC50s ranged from 0.125 to 16 micrograms/ml. The MIC90s were lowest with imipenem at 0.5 micrograms/ml, followed by cefoxitin at 32 micrograms/ml; they were highest with cefotetan at 128 micrograms/ml and were 64 micrograms/ml with the others. In vitro drug activity varied with the antibiotic, the organism, the method used, and the breakpoint selected. Rates of resistance varied considerably between the taxonomic groups of organisms tested and also among species within a group. Overall, reproducibility with the agar dilution method ranged from 44% to 85%; testing with ceftizoxime was the least reproducible. Microdilution results agreed within +/- 1 dilution of the agar dilution mode 79% to 95% of the time, with some variation between drugs and organisms tested. Because there were distinct differences in the activity of some drugs against certain species, no antibiotic can substitute for others in in vitro testing.

Discharge rate measurements in a canal using radiotracer methods.

PubMed

Pant, H J; Goswami, Sunil; Biswal, Jayashree; Samantray, J S; Sharma, V K

2016-06-01

Discharge rates of water were measured in a canal using radiotracer methods with an objective to validate the efficacy of Concrete Volute Pumps (CVPs) installed at various pumping stations along the canal. Pulse velocity and dilution methods were applied to measure the discharge rates using Iodine-131 as a radiotracer. The discharge rate measured in one of the sections of the canal using the pulse velocity method was found to be 22.5m(3)/s, whereas the discharge rates measured using the dilution method in four different sections of the canal varied from 20.27 to 20.62m(3)/s with single CVP in operation. The standard error in discharge rate measurements using dilution method ranged from ±1.1 to ±1.8%. The experimentally measured values of the discharge rate were in good agreement with the design value of the discharge rate (20m(3)/s) thus validating the performance of the CVPs used in the canal. Copyright © 2016 Elsevier Ltd. All rights reserved.

Dilution testing using rapid diagnostic tests in a HIV diagnostic algorithm: a novel alternative for confirmation testing in resource limited settings.

PubMed

Shanks, Leslie; Siddiqui, M Ruby; Abebe, Almaz; Piriou, Erwan; Pearce, Neil; Ariti, Cono; Masiga, Johnson; Muluneh, Libsework; Wazome, Joseph; Ritmeijer, Koert; Klarkowski, Derryck

2015-05-14

Current WHO testing guidelines for resource limited settings diagnose HIV on the basis of screening tests without a confirmation test due to cost constraints. This leads to a potential risk of false positive HIV diagnosis. In this paper, we evaluate the dilution test, a novel method for confirmation testing, which is simple, rapid, and low cost. The principle of the dilution test is to alter the sensitivity of a rapid diagnostic test (RDT) by dilution of the sample, in order to screen out the cross reacting antibodies responsible for falsely positive RDT results. Participants were recruited from two testing centres in Ethiopia where a tiebreaker algorithm using 3 different RDTs in series is used to diagnose HIV. All samples positive on the initial screening RDT and every 10th negative sample underwent testing with the gold standard and dilution test. Dilution testing was performed using Determine™ rapid diagnostic test at 6 different dilutions. Results were compared to the gold standard of Western Blot; where Western Blot was indeterminate, PCR testing determined the final result. 2895 samples were recruited to the study. 247 were positive for a prevalence of 8.5 % (247/2895). A total of 495 samples underwent dilution testing. The RDT diagnostic algorithm misclassified 18 samples as positive. Dilution at the level of 1/160 was able to correctly identify all these 18 false positives, but at a cost of a single false negative result (sensitivity 99.6 %, 95 % CI 97.8-100; specificity 100 %, 95 % CI: 98.5-100). Concordance between the gold standard and the 1/160 dilution strength was 99.8 %. This study provides proof of concept for a new, low cost method of confirming HIV diagnosis in resource-limited settings. It has potential for use as a supplementary test in a confirmatory algorithm, whereby double positive RDT results undergo dilution testing, with positive results confirming HIV infection. Negative results require nucleic acid testing to rule out false negative results due to seroconversion or misclassification by the lower sensitivity dilution test. Further research is needed to determine if these results can be replicated in other settings. ClinicalTrials.gov, NCT01716299 .

Method for processing aqueous wastes

DOEpatents

Pickett, John B.; Martin, Hollis L.; Langton, Christine A.; Harley, Willie W.

1993-01-01

A method for treating waste water such as that from an industrial processing facility comprising the separation of the waste water into a dilute waste stream and a concentrated waste stream. The concentrated waste stream is treated chemically to enhance precipitation and then allowed to separate into a sludge and a supernate. The supernate is skimmed or filtered from the sludge and blended with the dilute waste stream to form a second dilute waste stream. The sludge remaining is mixed with cementitious material, rinsed to dissolve soluble components, then pressed to remove excess water and dissolved solids before being allowed to cure. The dilute waste stream is also chemically treated to decompose carbonate complexes and metal ions and then mixed with cationic polymer to cause the precipitated solids to flocculate. Filtration of the flocculant removes sufficient solids to allow the waste water to be discharged to the surface of a stream. The filtered material is added to the sludge of the concentrated waste stream. The method is also applicable to the treatment and removal of soluble uranium from aqueous streams, such that the treated stream may be used as a potable water supply.

Evaluation of the pulse-contour method of determining stroke volume in man.

NASA Technical Reports Server (NTRS)

Alderman, E. L.; Branzi, A.; Sanders, W.; Brown, B. W.; Harrison, D. C.

1972-01-01

The pulse-contour method for determining stroke volume has been employed as a continuous rapid method of monitoring the cardiovascular status of patients. Twenty-one patients with ischemic heart disease and 21 patients with mitral valve disease were subjected to a variety of hemodynamic interventions. The pulse-contour estimations, using three different formulas derived by Warner, Kouchoukos, and Herd, were compared with indicator-dilution outputs. A comparison of the results of the two methods for determining stroke volume yielded correlation coefficients ranging from 0.59 to 0.84. The better performing Warner formula yielded a coefficient of variation of about 20%. The type of hemodynamic interventions employed did not significantly affect the results using the pulse-contour method. Although the correlation of the pulse-contour and indicator-dilution stroke volumes is high, the coefficient of variation is such that small changes in stroke volume cannot be accurately assessed by the pulse-contour method. However, the simplicity and rapidity of this method compared to determination of cardiac output by Fick or indicator-dilution methods makes it a potentially useful adjunct for monitoring critically ill patients.

Vitamins D and A can be successfully measured by LC-MS/MS in cord blood diluted plasma.

PubMed

Albarhani, Ali A; Collier, Fiona; Greaves, Ronda F; Ponsonby, Anne-Louise; Allen, Katrina J; Vuillermin, Peter J; Roche, Peter; Clarke, Michael W

2015-11-01

In widely used protocols for the collection and isolation of cord blood mononuclear cells, investigators are left with substantial volumes of diluted plasma which could be used for other measurements. The aim of this study was to ascertain the validity of umbilical cord blood (UCB) diluted plasma samples for vitamin D, A and E analysis compared to UCB serum samples. Twenty UCB matched samples of diluted plasma and serum were collected. The samples were analysed by two liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods on two separate occasions. The results of 25(OH)D3 obtained by the two laboratories demonstrated close agreement with a mean difference of 0.14nmol/L [95% confidence interval (95% CI), -6.8 to 7.1]. Both methods demonstrate close agreement for 25(OH)D3 in UCB serum versus diluted UCB plasma; mean difference 2.2nmol/L [95% CI, -9.5 to 13.9] and 4.1nmol/L [95% CI, -14.5 to 6.1] for the results from Lab A and Lab B, respectively. Vitamin A was quantified by Lab A in UCB serum and diluted UCB plasma; mean difference 0.07μmol/L [95% CI, -0.41 to 0.28]. Results of 25(OH)D3 epimer and vitamin E in the diluted UCB plasma were below the limit of quantification, and could not be compared with UCB serum. Diluted UCB plasma can be used for the quantification of retinol and 25(OH)D3 by LC-MS/MS. By contrast, quantification of 25(OH)D3 epimer and vitamin E in diluted UCB plasma is not supported by this study due to limitations in analytical sensitivity. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Effectiveness of qPCR permutations, internal controls and dilution as means for minimizing the impact of inhibition while measuring Enterococcus in environmental waters.

PubMed

Cao, Y; Griffith, J F; Dorevitch, S; Weisberg, S B

2012-07-01

  Draft criteria for the optional use of qPCR for recreational water quality monitoring have been published in the United States. One concern is that inhibition of the qPCR assay can lead to false-negative results and potentially inadequate public health protection. We evaluate the effectiveness of strategies for minimizing the impact of inhibition.   Five qPCR method permutations for measuring Enterococcus were challenged with 133 potentially inhibitory fresh and marine water samples. Serial dilutions were conducted to assess Enterococcus target assay inhibition, to which inhibition identified using four internal controls (IC) was compared. The frequency and magnitude of inhibition varied considerably among qPCR methods, with the permutation using an environmental master mix performing substantially better. Fivefold dilution was also effective at reducing inhibition in most samples (>78%). ICs were variable and somewhat ineffective, with 54-85% agreement between ICs and serial dilution.   The current IC methods appear to not accurately predict Enterococcus inhibition and should be used with caution; fivefold dilution and the use of reagents designed for environmental sample analysis (i.e. more robust qPCR chemistry) may be preferable.   Suitable approaches for defining, detecting and reducing inhibition will improve implementation of qPCR for water monitoring. © 2012 The Authors. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

Investigation of Possible Maillard Reaction Between Acyclovir and Dextrose upon Dilution Prior to Parenteral Administration.

PubMed

Siahi Shadbad, Mohammad Reza; Ghaderi, Faranak; Hatami, Leila; Monajjemzadeh, Farnaz

2016-12-01

In this study the stability of parenteral acyclovir (ACV) when diluted in dextrose (DEX) as large volume intravenous fluid preparation (LVIF) was evaluated and the possible Maillard reaction adducts were monitored in the recommended infusion time. Different physicochemical methods were used to evaluate the Maillard reaction of dextrose with ACV to track the reaction in real infusion condition. Other large volume intravenous fluids were checked regarding the diluted drug stability profile. Differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), and mass data proved the reaction of glucose with dextrose. A Maillard-specific high performance liquid chromatography (HPLC) method was used to track the reaction in real infusion condition in vitro. The nucleophilic reaction occurred in diluted parenteral preparations of acyclovir in 5% dextrose solutions. The best diluent solution was also selected as sodium chloride and introduced based on drug stability and also its adsorption onto different infusion sets (PVC or non PVC) to provide an acceptable administration protocol in clinical practices. Although, the Maillard reaction was proved and successfully tracked in diluted solutions, and the level of drug loss when diluted in dextrose was reported to be between 0.27 up to 1.03% of the initial content. There was no drug adsorption to common infusion sets. The best diluent for parenteral acyclovir is sodium chloride large volume intravenous fluid.

Quantitative Analysis by Isotopic Dilution Using Mass Spectroscopy: The Determination of Caffeine by GC-MS.

ERIC Educational Resources Information Center

Hill, Devon W.; And Others

1988-01-01

Describes a laboratory technique for quantitative analysis of caffeine by an isotopic dilution method for coupled gas chromatography-mass spectroscopy. Discusses caffeine analysis and experimental methodology. Lists sample caffeine concentrations found in common products. (MVL)

Microfluidic Serial Dilution Circuit

PubMed Central

Paegel, Brian M.; Grover, William H.; Skelley, Alison M.; Mathies, Richard A.; Joyce, Gerald F.

2008-01-01

In vitro evolution of RNA molecules requires a method for executing many consecutive serial dilutions. To solve this problem, a microfluidic circuit has been fabricated in a three-layer glass-PDMS-glass device. The 400-nL serial dilution circuit contains five integrated membrane valves: three two-way valves arranged in a loop to drive cyclic mixing of the diluent and carryover, and two bus valves to control fluidic access to the circuit through input and output channels. By varying the valve placement in the circuit, carryover fractions from 0.04 to 0.2 were obtained. Each dilution process, which is comprised of a diluent flush cycle followed by a mixing cycle, is carried out with no pipeting, and a sample volume of 400 nL is sufficient for conducting an arbitrary number of serial dilutions. Mixing is precisely controlled by changing the cyclic pumping rate, with a minimum mixing time of 22 s. This microfluidic circuit is generally applicable for integrating automated serial dilution and sample preparation in almost any microfluidic architecture. PMID:17073422

Evaluation and reporting of enzyme immunoassay determinations of antibody to herpes simplex virus in sera and cerebrospinal fluid.

PubMed Central

Cremer, N E; Cossen, C K; Hanson, C V; Shell, G R

1982-01-01

Several methods for evaluating and reporting enzyme immunoassay (EIA) determinations of antibody to herpes simplex virus derived from one dilution of single serum samples were studied. An EIA ratio method for serological evidence of current infection from paired serum samples was also evaluated. Optical density (OD) of the reaction at a 1:100 serum dilution and estimated titers obtained by reference of the OD of the serum dilution to a standard curve were compared to the corresponding plotted EIA titer obtained by titration to endpoint. Neither the OD per se nor the estimated titer was completely predictive of the plotted titer (correlation coefficient [r] of 0.824 and 0.817, respectively), and they provided only a semiquantitative measurement of antibody concentration. For an antibody status report, however, OD would be sufficient if related to the cutoff value as an EIA index (OD of sample divided by cutoff OD for positive specimens). The OD of the EIA reaction at a single dilution (1:5) of cerebrospinal fluid, on the other hand, correlated quite well with the titer obtained by titration (r = 0.950). For serological diagnosis of current infection, the OD ratio of convalescence-phase/acute-phase sera was determined at several dilutions. A ratio of greater than or equal to 1.54 was calculated as a reliable index for a significant rise in antibody concentration and compatible with current infection. By determining the convalescent-phase/acute-phase serum ratio at two dilutions, 1:100 and 1:1,000, the EIA ratio method appeared to be a sensitive as or more sensitive than, complement fixation in diagnosing current infection. PMID:6284791

Effect of mixing method on the mixing degree during the preparation of triturations.

PubMed

Nakamura, Hitoshi; Yanagihara, Yoshitsugu; Sekiguchi, Hiroko; Komada, Fusao; Kawabata, Haruno; Ohtani, Michiteru; Saitoh, Yukiya; Kariya, Satoru; Suzuki, Hiroshi; Uchino, Katsuyoshi; Iga, Tatsuji

2004-03-01

By using lactose colored with erythrocin, we investigated the effects of mixing methods on mixing degree during the preparation of trituration with a mortar and pestle. The extent of powder dilution was set to 4 to 64 fold in the experiments. We compared the results obtained by using two methods: (1) one-step mixing of powders after addition of diluents and (2) gradual mixing of powders after addition of diluents. As diluents, we used crystallized lactose and powdered lactose for the preparation of trituration. In the preparation of 64-fold trituration, an excellent degree of mixing was obtained, with CV values of less than 6.08%, for both preparation methods and for the two kinds of diluents. The mixing of two kinds of powders whose distributions of particle sizes were similar resulted in much better degree of mixing, with CV values of less than 3.0%. However, the concentration of principal agents in 64-fold trituration was reduced by 20% due to the adsorption of dye to the apparatus. Under conditions in which a much higher dilution rate and/or much better degree of dilution was required, it must be necessary to dilute powders with considering their physicality and to determine the concentrations of principal agents after the mixing.

A new serotyping method for Klebsiella species: development of the technique.

PubMed Central

Riser, E; Noone, P; Poulton, T A

1976-01-01

A new serotyping method for Klebsiella species using indirect immunofluorescence is described. Nonspecific fluorescence has been minimized by carrying out the capsular antigen-antibody reaction at pH 9.0. Commercial antisera have been tested with the 72 antigenic types of Klebsiella, and appropriate dilutions of each pool and specific antisera have been proposed for use in routine typing. Dilutions were chosen to allow strong fluorescence with each type and its specific antiserum and minimal fluorescence with cross reacting antisera. Where the pool antisera gave a weak reaction for one or more of the component types, it is recommended that the specific antisera for these types be added to the pool dilution. The few remaining cross reactions, with the pool and specific antisera in test dilution, are listed in a table. The unique cross reacting patterns of particular types have been found to be useful in identification. Typing Klebsiella by the fluorescent antibody technique is easy to perform and interpret; the results are reproducible, and it is less expensive than the existing capsular swelling method as it is more sensitive and requires less concentrated antisera. This new method of typing should facilitate detailed epidemiological studies of the mode of transmission of Klebsiella species in hospitals and thus allow more effective infection control measures to be instituted. Images PMID:777042

Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis.

PubMed

Alhassan, Andy; Govind, Yadav; Tam, Nguyen Thanh; Thekisoe, Oriel M M; Yokoyama, Naoaki; Inoue, Noboru; Igarashi, Ikuo

2007-04-01

The sensitivity of LAMP, PCR and in vitro culture methods for the detection of Theileria equi and Babesia caballi was evaluated using tenfold serially diluted culture parasites. On day 1 post-culture, both T. equi and B. caballi parasites could only be observed at 1% parasite dilution from the in vitro culture method, whereas LAMP could detect up to 1 x 10(-3)% of both T. equi and B. caballi parasite dilutions, whilst PCR could detect 1 x 10(-3)% T. equi and 1 x 10(-1)% B. caballi parasite dilutions. On day 7 post-culture, the detection limit for T. equi and B. caballi in the in vitro culture increased up to 1 x 10(-6)%, whereas LAMP detection limit increased to 1 x 10(-10)% for both parasites, whilst the PCR detection limit increased to 1 x 10(-10)% and 1 x 10(-6)% for T. equi and B. caballi, respectively. Furthermore, LAMP and PCR amplified the T. equi DNA extracted from the organs of an experimentally infected horse. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of equine piroplasmosis and together with PCR can also be used as supplementary methods during post-mortems.

Fast and accurate calculation of dilute quantum gas using Uehling–Uhlenbeck model equation

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

Yano, Ryosuke, E-mail: ryosuke.yano@tokiorisk.co.jp

The Uehling–Uhlenbeck (U–U) model equation is studied for the fast and accurate calculation of a dilute quantum gas. In particular, the direct simulation Monte Carlo (DSMC) method is used to solve the U–U model equation. DSMC analysis based on the U–U model equation is expected to enable the thermalization to be accurately obtained using a small number of sample particles and the dilute quantum gas dynamics to be calculated in a practical time. Finally, the applicability of DSMC analysis based on the U–U model equation to the fast and accurate calculation of a dilute quantum gas is confirmed by calculatingmore » the viscosity coefficient of a Bose gas on the basis of the Green–Kubo expression and the shock layer of a dilute Bose gas around a cylinder.« less

Reusable chelating resins concentrate metal ions from highly dilute solutions

NASA Technical Reports Server (NTRS)

Bauman, A. J.; Weetal, H. H.; Weliky, N.

1966-01-01

Column chromatographic method uses new metal chelating resins for recovering heavy-metal ions from highly dilute solutions. The absorbed heavy-metal cations may be removed from the chelating resins by acid or base washes. The resins are reusable after the washes are completed.

Study the Effect of SiO2 Based Flux on Dilution in Submerged Arc Welding

NASA Astrophysics Data System (ADS)

kumar, Aditya; Maheshwari, Sachin

2017-08-01

This paper highlights the method for prediction of dilution in submerged arc welding (SAW). The most important factors of weld bead geometry are governed by the weld dilution which controls the chemical and mechanical properties. Submerged arc welding process is used generally due to its very easy control of process variables, good penetration, high weld quality, and smooth finish. Machining parameters, with suitable weld quality can be achieved with the different composition of the flux in the weld. In the present study Si02-Al2O3-CaO flux system was used. In SiO2 based flux NiO, MnO, MgO were mixed in various proportions. The paper investigates the relationship between the process parameters like voltage, % of flux constituents and dilution with the help of Taguchi’s method. The experiments were designed according to Taguchi L9 orthogonal array, while varying the voltage at two different levels in addition to alloying elements. Then the optimal results conditions were verified by confirmatory experiments.

Breakdown of cell wall nanostructure in dilute acid pretreated biomass.

PubMed

Pingali, Sai Venkatesh; Urban, Volker S; Heller, William T; McGaughey, Joseph; O'Neill, Hugh; Foston, Marcus; Myles, Dean A; Ragauskas, Arthur; Evans, Barbara R

2010-09-13

The generation of bioethanol from lignocellulosic biomass holds great promise for renewable and clean energy production. A better understanding of the complex mechanisms of lignocellulose breakdown during various pretreatment methods is needed to realize this potential in a cost and energy efficient way. Here we use small-angle neutron scattering (SANS) to characterize morphological changes in switchgrass lignocellulose across molecular to submicrometer length scales resulting from the industrially relevant dilute acid pretreatment method. Our results demonstrate that dilute acid pretreatment increases the cross-sectional radius of the crystalline cellulose fibril. This change is accompanied by removal of hemicellulose and the formation of R(g) ∼ 135 A lignin aggregates. The structural signature of smooth cell wall surfaces is observed at length scales larger than 1000 A, and it remains remarkably invariable during pretreatment. This study elucidates the interplay of the different biomolecular components in the breakdown process of switchgrass by dilute acid pretreatment. The results are important for the development of efficient strategies of biomass to biofuel conversion.

Infinitely dilute partial molar properties of proteins from computer simulation.

PubMed

Ploetz, Elizabeth A; Smith, Paul E

2014-11-13

A detailed understanding of temperature and pressure effects on an infinitely dilute protein's conformational equilibrium requires knowledge of the corresponding infinitely dilute partial molar properties. Established molecular dynamics methodologies generally have not provided a way to calculate these properties without either a loss of thermodynamic rigor, the introduction of nonunique parameters, or a loss of information about which solute conformations specifically contributed to the output values. Here we implement a simple method that is thermodynamically rigorous and possesses none of the above disadvantages, and we report on the method's feasibility and computational demands. We calculate infinitely dilute partial molar properties for two proteins and attempt to distinguish the thermodynamic differences between a native and a denatured conformation of a designed miniprotein. We conclude that simple ensemble average properties can be calculated with very reasonable amounts of computational power. In contrast, properties corresponding to fluctuating quantities are computationally demanding to calculate precisely, although they can be obtained more easily by following the temperature and/or pressure dependence of the corresponding ensemble averages.

Method for processing aqueous wastes

DOEpatents

Pickett, J.B.; Martin, H.L.; Langton, C.A.; Harley, W.W.

1993-12-28

A method is presented for treating waste water such as that from an industrial processing facility comprising the separation of the waste water into a dilute waste stream and a concentrated waste stream. The concentrated waste stream is treated chemically to enhance precipitation and then allowed to separate into a sludge and a supernate. The supernate is skimmed or filtered from the sludge and blended with the dilute waste stream to form a second dilute waste stream. The sludge remaining is mixed with cementitious material, rinsed to dissolve soluble components, then pressed to remove excess water and dissolved solids before being allowed to cure. The dilute waste stream is also chemically treated to decompose carbonate complexes and metal ions and then mixed with cationic polymer to cause the precipitated solids to flocculate. Filtration of the flocculant removes sufficient solids to allow the waste water to be discharged to the surface of a stream. The filtered material is added to the sludge of the concentrated waste stream. The method is also applicable to the treatment and removal of soluble uranium from aqueous streams, such that the treated stream may be used as a potable water supply. 4 figures.

In situ formation of phosphate barriers in soil

DOEpatents

Moore, Robert C.

2002-01-01

Reactive barriers and methods for making reactive barriers in situ in soil for sequestering soil ontaminants including actinides and heavy metals. The barrier includes phosphate, and techniques are disclosed for forming specifically apatite barriers. The method includes injecting dilute reagents into soil in proximity to a contamination plume or source such as a waste drum to achieve complete or partial encapsulation of the waste. Controlled temperature and pH facilitates rapid formation of apatite, for example, where dilute aqueous calcium chloride and dilute aqueous sodium phosphate are the selected reagents. Mixing of reagents to form precipitate is mediated and enhanced through movement of reagents in soil as a result of phenomena including capillary action, movement of groundwater, soil washing and reagent injection pressure.

Fecal electrolyte testing for evaluation of unexplained diarrhea: Validation of body fluid test accuracy in the absence of a reference method.

PubMed

Voskoboev, Nikolay V; Cambern, Sarah J; Hanley, Matthew M; Giesen, Callen D; Schilling, Jason J; Jannetto, Paul J; Lieske, John C; Block, Darci R

2015-11-01

Validation of tests performed on body fluids other than blood or urine can be challenging due to the lack of a reference method to confirm accuracy. The aim of this study was to evaluate alternate assessments of accuracy that laboratories can rely on to validate body fluid tests in the absence of a reference method using the example of sodium (Na(+)), potassium (K(+)), and magnesium (Mg(2+)) testing in stool fluid. Validations of fecal Na(+), K(+), and Mg(2+) were performed on the Roche cobas 6000 c501 (Roche Diagnostics) using residual stool specimens submitted for clinical testing. Spiked recovery, mixing studies, and serial dilutions were performed and % recovery of each analyte was calculated to assess accuracy. Results were confirmed by comparison to a reference method (ICP-OES, PerkinElmer). Mean recoveries for fecal electrolytes were Na(+) upon spiking=92%, mixing=104%, and dilution=105%; K(+) upon spiking=94%, mixing=96%, and dilution=100%; and Mg(2+) upon spiking=93%, mixing=98%, and dilution=100%. When autoanalyzer results were compared to reference ICP-OES results, Na(+) had a slope=0.94, intercept=4.1, and R(2)=0.99; K(+) had a slope=0.99, intercept=0.7, and R(2)=0.99; and Mg(2+) had a slope=0.91, intercept=-4.6, and R(2)=0.91. Calculated osmotic gap using both methods were highly correlated with slope=0.95, intercept=4.5, and R(2)=0.97. Acid pretreatment increased magnesium recovery from a subset of clinical specimens. A combination of mixing, spiking, and dilution recovery experiments are an acceptable surrogate for assessing accuracy in body fluid validations in the absence of a reference method. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Reverse isotope dilution method for determining benzene and metabolites in tissues

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

Bechtold, W.E.; Sabourin, P.J.; Henderson, R.F.

1988-07-01

A method utilizing reverse isotope dilution for the analysis of benzene and its organic soluble metabolites in tissues of rats and mice is presented. Tissues from rats and mice that had been exposed to radiolabeled benzene were extracted with ethyl acetate containing known, excess quantities of unlabeled benzene and metabolites. Butylated hydroxytoluene was added as an antioxidant. The ethyl acetate extracts were analyzed with semipreparative reversed-phase HPLC. Isolated peaks were collected and analyzed for radioactivity (by liquid scintillation spectrometry) and for mass (by UV absorption). The total amount of each compound present was calculated from the mass dilution of themore » radiolabeled isotope. This method has the advantages of high sensitivity, because of the high specific activity of benzene, and relative stability of the analyses, because of the addition of large amounts of unlabeled carrier analogue.« less

Rapid Detection of Methicillin-Resistant Staphylococcus aureus Isolates by Turanose Fermentation Method

PubMed Central

Raeisi, Javad; Saifi, Mahnaz; Pourshafie, Mohammad Reza; Asadi Karam, Mohammad Reza; Mohajerani, Hamid Reza

2015-01-01

Background: Methicillin-Resistant Staphylococcus aureus (MRSA) is a major pathogen in the hospital and community settings. Rapid methods to diagnose S. aureus infections are sought by many researchers worldwide. The current study aimed to utilize a phenotypic method of turanose fermentation to identify methicillin-susceptible and resistant S. aureus. Objectives: The current study aimed to assay the turanose metabolism at different dilutions as a rapid phenotypic method to identify MRSA isolates. Materials and Methods: A total of 150 Staphylococcus isolates were collected from Tehran health centers. Staphylococcus aureus isolates were identified based on cultural characteristics, biochemical reactions and positive tube coagulase test. Methicillin resistance was determined by the disk diffusion method. The Polymerase Chain Reaction amplification was used to detect the mecA gene in MRSA isolates. All the methicillin-resistant and susceptible isolates were evaluated for turanose metabolism with 1%, 0.7% and 0.5% dilutions using the microplate method. Results: Out of the 150 staphylococcal isolates, 80 were identified as S. aureus. Among which 40 (50%) of the isolates were MRSA. The mecA gene was present in all S. aureus isolates resistant to methicillin. A considerable difference was also observed between susceptible and resistant isolates of S. aureus at a 0.7% dilution of turanose. Conclusions: Since it is highly important to rapidly detect MRSA isolates, especially in nosocomial infections, phenotypic methods may certainly be useful for this purpose. Resistance to methicillin in S. aureus shows a substantially increased ability in turanose metabolism. It is concluded that fermentation of turanose at 0.7% dilution could be a rapid detection method for primary screening of MRSA isolates. PMID:26495105

Determination of diffusion coefficients of various livestock antibiotics in water at infinite dilution

NASA Astrophysics Data System (ADS)

Soriano, Allan N.; Adamos, Kristoni G.; Bonifacio, Pauline B.; Adornado, Adonis P.; Bungay, Vergel C.; Vairavan, Rajendaran

2017-11-01

The fate of antibiotics entering the environment raised concerns on the possible effect of antimicrobial resistance bacteria. Prediction of the fate and transport of these particles are needed to be determined, significantly the diffusion coefficient of antibiotic in water at infinite dilution. A systematic determination of diffusion coefficient of antibiotic in water at infinite dilution of five different kinds of livestock antibiotics namely: Amtyl, Ciprotyl, Doxylak Forte, Trisullak, and Vetracin Gold in the 293.15 to 313.15 K temperature range are reported through the use of the method involving the electrolytic conductivity measurements. A continuous stirred tank reactor is utilized to measure the electrolytic conductivities of the considered systems. These conductivities are correlated by using the Nernst-Haskell equation to determine the infinite dilution diffusion coefficient. Determined diffusion coefficients are based on the assumption that in dilute solution, these antibiotics behave as strong electrolyte from which H+ cation dissociate from the antibiotic's anion.

Enhanced Detection of Vibrio Cholerae in Oyster Homogenate Based on Centrifugal Removal of Inhibitory Agents

NASA Technical Reports Server (NTRS)

Alexander, Donita; DePaola, Angelo; Young, Ronald B.

1998-01-01

The disease cholera, caused by Vibrio cholerae, has been associated with consumption of contaminated seafood, including raw oysters. Detection of V. cholerae in foods typically involves blending the oysters, diluting the homogenate in alkaline peptone water (APW), overnight enrichment, and isolation on selective agar. Unfortunately, the oyster homogenate must be diluted to large volumes because lower dilutions inhibit the growth of V. cholerae. The goals of this study were to develop an alternative to large dilutions and to evaluate the basis for the inhibition observed in lower dilutions of oyster homogenates. Centrifugation of oyster homogenates at 10,000 x g for 15 min, followed by enrichment of the resulting pellet in APW, was found to eliminate the inhibition of V. cholerae growth. Inhibition appears not to be due to competing microflora but to a component(s) released when V. cholerae grows in the presence of oyster homogenate. The inhibitory component(s) kills the V. cholerae after the cell concentration reaches > 10(exp 8) cells/mL, rather than initially preventing their growth. The pH also declines from 8.0 to 5.5 during this period; however, the pH decline by itself appears not to cause V. cholerae death. Seven strains of V. cholerae (01 and non-01) and two strains of V. vulnificus were susceptible to the inhibitory agent(s). However, other Vibrio and non-Vibrio species tested were not inhibited by the oyster homogenates. Based on digestion of oyster homogenates with pronase, trypsin and lipase, the inhibitory reaction involves a protein(s). In a preliminary trial with oyster homogenate seeded with 1 cfu/g of V. cholerae, the modified centrifugation technique detected a slightly higher percentage of samples at a 1:10 dilution than the standard FDA Bacteriological Analytical Method (BAM) detected in uncentrifuged oyster homogenate at a 1:100 dilution. V. cholerae in seeded samples could also be detected more frequently by the modified centrifugation method than by PCR at a 1:10 dilution.

A mass-balance code for the quantitative interpretation of fluid column profiles in ground-water studies

NASA Astrophysics Data System (ADS)

Paillet, Frederick

2012-08-01

A simple mass-balance code allows effective modeling of conventional fluid column resistivity logs in dilution tests involving column replacement with either distilled water or dilute brine. Modeling a series of column profiles where the inflowing formation water introduces water quality interfaces propagating along the borehole gives effective estimates of the rate of borehole flow. Application of the dilution model yields estimates of borehole flow rates that agree with measurements made with the heat-pulse flowmeter under ambient and pumping conditions. Model dilution experiments are used to demonstrate how dilution logging can extend the range of borehole flow measurement at least an order of magnitude beyond that achieved with flowmeters. However, dilution logging has the same dynamic range limitation encountered with flowmeters because it is difficult to detect and characterize flow zones that contribute a small fraction of total flow when that contribution is superimposed on a larger flow. When the smaller contribution is located below the primary zone, ambient downflow may disguise the zone if pumping is not strong enough to reverse the outflow. This situation can be addressed by increased pumping. But this is likely to make the moveout of water quality interfaces too fast to measure in the upper part of the borehole, so that a combination of flowmeter and dilution method may be more appropriate. Numerical experiments show that the expected weak horizontal flow across the borehole at conductive zones would be almost impossible to recognize if any ambient vertical flow is present. In situations where natural water quality differences occur such as flowing boreholes or injection experiments, the simple mass-balance code can be used to quantitatively model the evolution of fluid column logs. Otherwise, dilution experiments can be combined with high-resolution flowmeter profiles to obtain results not attainable using either method alone.

Community-Level Physiological Profiling of Microbial Communities in Constructed Wetlands: Effects of Sample Preparation.

PubMed

Button, Mark; Weber, Kela; Nivala, Jaime; Aubron, Thomas; Müller, Roland Arno

2016-03-01

Community-level physiological profiling (CLPP) using BIOLOG® EcoPlates™ has become a popular method for characterizing and comparing the functional diversity, functional potential, and metabolic activity of heterotrophic microbial communities. The method was originally developed for profiling soil communities; however, its usage has expanded into the fields of ecotoxicology, agronomy, and the monitoring and profiling of microbial communities in various wastewater treatment systems, including constructed wetlands for water pollution control. When performing CLPP on aqueous samples from constructed wetlands, a wide variety of sample characteristics can be encountered and challenges may arise due to excessive solids, color, or turbidity. The aim of this study was to investigate the impacts of different sample preparation methods on CLPP performed on a variety of aqueous samples covering a broad range of physical and chemical characteristics. The results show that using filter paper, centrifugation, or settling helped clarify samples for subsequent CLPP analysis, however did not do so as effectively as dilution for the darkest samples. Dilution was able to provide suitable clarity for the darkest samples; however, 100-fold dilution significantly affected the carbon source utilization patterns (CSUPs), particularly with samples that were already partially or fully clear. Ten-fold dilution also had some effect on the CSUPs of samples which were originally clear; however, the effect was minimal. Based on these findings, for this specific set of samples, a 10-fold dilution provided a good balance between ease of use, sufficient clarity (for dark samples), and limited effect on CSUPs. The process and findings outlined here can hopefully serve future studies looking to utilize CLPP for functional analysis of microbial communities and also assist in comparing data from studies where different sample preparation methods were utilized.

An Isotopic Dilution Experiment Using Liquid Scintillation: A Simple Two-System, Two-Phase Analysis.

ERIC Educational Resources Information Center

Moehs, Peter J.; Levine, Samuel

1982-01-01

A simple isotonic, dilution analysis whose principles apply to methods of more complex radioanalyses is described. Suitable for clinical and instrumental analysis chemistry students, experimental manipulations are kept to a minimum involving only aqueous extraction before counting. Background information, procedures, and results are discussed.…

An improved method for undertaking limiting dilution assays for in vitro cloning of Plasmodium falciparum parasites.

PubMed

Butterworth, Alice S; Robertson, Alan J; Ho, Mei-Fong; Gatton, Michelle L; McCarthy, James S; Trenholme, Katharine R

2011-04-18

Obtaining single parasite clones is required for many techniques in malaria research. Cloning by limiting dilution using microscopy-based assessment for parasite growth is an arduous and labor-intensive process. An alternative method for the detection of parasite growth in limiting dilution assays is using a commercial ELISA histidine-rich protein II (HRP2) detection kit. Detection of parasite growth was undertaken using HRP2 ELISA and compared to thick film microscopy. An HRP2 protein standard was used to determine the detection threshold of the HRP2 ELISA assay, and a HRP2 release model was used to extrapolate the amount of parasite growth required for a positive result. The HRP2 ELISA was more sensitive than microscopy for detecting parasite growth. The minimum level of HRP2 protein detection of the ELISA was 0.11 ng/ml. Modeling of HRP2 release determined that 2,116 parasites are required to complete a full erythrocytic cycle to produce sufficient HRP2 to be detected by the ELISA. Under standard culture conditions this number of parasites is likely to be reached between 8 to 14 days of culture. This method provides an accurate and simple way for the detection of parasite growth in limiting dilution assays, reducing time and resources required in traditional methods. Furthermore the method uses spent culture media instead of the parasite-infected red blood cells, enabling culture to continue. © 2011 Butterworth et al; licensee BioMed Central Ltd.

Methods of preventing vinorelbine-induced phlebitis: an experimental study in rabbits

PubMed Central

Kohno, Emiko; Murase, Saori; Nishikata, Mayumi; Okamura, Noboru; Matzno, Sumio; Kuwahara, Takashi; Matsuyama, Kenji

2008-01-01

Purpose: In order to identify methods for preventing phlebitis caused by intravenous administration of vinorelbine (VNR), we established a procedure for estimating the severity of phlebitis in an animal model. Methods: Four different factors (administration rate, dilution, flushing, and infusion of fat emulsion) were evaluated for alleviation of phlebitis caused by VNR infusion. VNR was diluted with normal saline to prepare test solutions with concentrations of 0.6 mg/mL or 0.3 mg/mL for infusion into the auricular veins of rabbits. Two days after VNR infusion, the veins were subjected to histopathological examination. Results: VNR did not cause obvious loss of venous endothelial cells, the most sensitive and common feature of phlebitis, but VNR infusion led to inflammatory cell infiltration, edema, and epidermal degeneration. Tissue damage was significantly decreased by shortening the administration time and by diluting the VNR solution for infusion from 0.6 mg/mL to 0.3 mg/mL. However, there was no effect of flushing with normal saline after VNR infusion, while treatment with fat emulsion before and after VNR infusion only had a minimal effect. Conclusion: Rapid infusion and dilution are effective methods of reducing phlebitis caused by the infusion of VNR, but the efficacy of flushing with normal saline or infusion of fat emulsion was not confirmed. PMID:18695742

Improvement of pre-treatment method for 36Cl/Cl measurement of Cl in natural groundwater by AMS

NASA Astrophysics Data System (ADS)

Nakata, Kotaro; Hasegawa, Takuma

2011-02-01

Estimation of 36Cl/Cl by accelerator mass spectrometry (AMS) is a useful method to trace hydrological processes in groundwater. For accurate estimation, separation of SO42- from Cl - in groundwater is required because 36S affects AMS measurement of 36Cl. Previous studies utilized the difference in solubility between BaSO 4 and BaCl 2 (BaSO 4 method) to chemically separate SO42- from Cl -. However, the accuracy of the BaSO 4 method largely depends on operator skill, and consequently Cl - recovery is typically incomplete (70-80%). In addition, the method is time consuming (>1 week), and cannot be applied directly to dilute solutions. In this study, a method based on ion-exchange column chromatography (column method) was developed for separation of Cl - and SO42-. Optimum conditions were determined for the diameter and height of column, type and amount of resin, type and concentration of eluent, and flow rate. The recovery of Cl - was almost 100%, which allowed complete separation from SO42-. The separation procedure was short (<6 h), and was successfully applied to dilute (1 mg/L Cl) solution. Extracted pore water and diluted seawater samples were processed by the column and BaSO 4 methods, and then analyzed by AMS to estimate 36S counts and 36Cl/Cl values. 36S counts in samples processed by the column method were stable and lower than those from the BaSO 4 method. The column method has the following advantages over the BaSO 4 method: (1) complete and stable separation of Cl - and SO42-, (2) less operator influence on results, (3) short processing time (<6 h), (4) high (almost 100%) recovery of Cl -, and (5) concentration of Cl - and separation from SO42- in the one system for dilute solutions.

STATISTICAL ESTIMATES OF VARIANCE FOR 15N ISOTOPE DILUTION MEASUREMENTS OF GROSS RATES OF NITROGEN CYCLE PROCESSES

EPA Science Inventory

It has been fifty years since Kirkham and Bartholmew (1954) presented the conceptual framework and derived the mathematical equations that formed the basis of the now commonly employed method of 15N isotope dilution. Although many advances in methodology and analysis have been ma...

Methods of preventing vinorelbine-induced phlebitis: an experimental study in rabbits.

PubMed

Kohno, Emiko; Murase, Saori; Nishikata, Mayumi; Okamura, Noboru; Matzno, Sumio; Kuwahara, Takashi; Matsuyama, Kenji

2008-07-22

In order to identify methods for preventing phlebitis caused by intravenous administration of vinorelbine (VNR), we established a procedure for estimating the severity of phlebitis in an animal model. Four different factors (administration rate, dilution, flushing, and infusion of fat emulsion) were evaluated for alleviation of phlebitis caused by VNR infusion. VNR was diluted with normal saline to prepare test solutions with concentrations of 0.6 mg/mL or 0.3 mg/mL for infusion into the auricular veins of rabbits. Two days after VNR infusion, the veins were subjected to histopathological examination. VNR did not cause obvious loss of venous endothelial cells, the most sensitive and common feature of phlebitis, but VNR infusion led to inflammatory cell infiltration, edema, and epidermal degeneration. Tissue damage was significantly decreased by shortening the administration time and by diluting the VNR solution for infusion from 0.6 mg/mL to 0.3 mg/mL. However, there was no effect of flushing with normal saline after VNR infusion, while treatment with fat emulsion before and after VNR infusion only had a minimal effect. Rapid infusion and dilution are effective methods of reducing phlebitis caused by the infusion of VNR, but the efficacy of flushing with normal saline or infusion of fat emulsion was not confirmed.

Single point dilution method for the quantitative analysis of antibodies to the gag24 protein of HIV-1.

PubMed

Palenzuela, D O; Benítez, J; Rivero, J; Serrano, R; Ganzó, O

1997-10-13

In the present work a concept proposed in 1992 by Dopotka and Giesendorf was applied to the quantitative analysis of antibodies to the p24 protein of HIV-1 in infected asymptomatic individuals and AIDS patients. Two approaches were analyzed, a linear model OD = b0 + b1.log(titer) and a nonlinear log(titer) = alpha.OD beta, similar to the Dopotka-Giesendorf's model. The above two proposed models adequately fit the dependence of the optical density values at a single point dilution, and titers achieved by the end point dilution method (EPDM). Nevertheless, the nonlinear model better fits the experimental data, according to residuals analysis. Classical EPDM was compared with the new single point dilution method (SPDM) using both models. The best correlation between titers calculated using both models and titers achieved by EPDM was obtained with the nonlinear model. The correlation coefficients for the nonlinear and linear models were r = 0.85 and r = 0.77, respectively. A new correction factor was introduced into the nonlinear model and this reduced the day-to-day variation of titer values. In general, SPDM saves time, reagents and is more precise and sensitive to changes in antibody levels, and therefore has a higher resolution than EPDM.

Preparation of a silicon surface for subsequent growth of dilute nitride alloys by molecular-beam epitaxy

NASA Astrophysics Data System (ADS)

Lazarenko, A. A.; Berezovskaya, T. N.; Denisov, D. V.; Sobolev, M. S.; Pirogov, E. V.; Nikitina, E. V.

2017-11-01

This article discusses the process of preparation of a silicon surface for subsequent growth of dilute nitride alloys by molecular-beam epitaxy. The method of preparation of Si (100) and Si (111) substrates was developed. This method provides reproducible high-quality silicon surface for molecular-beam epitaxy of Si-GaP heterostructures. As a result, it managed to reduce the eviction oxide temperature below 800 °C, which is an important parameter for the MBE technology.

Mitochondrial DNA as a non-invasive biomarker: Accurate quantification using real time quantitative PCR without co-amplification of pseudogenes and dilution bias

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

Malik, Afshan N., E-mail: afshan.malik@kcl.ac.uk; Shahni, Rojeen; Rodriguez-de-Ledesma, Ana

2011-08-19

Highlights: {yields} Mitochondrial dysfunction is central to many diseases of oxidative stress. {yields} 95% of the mitochondrial genome is duplicated in the nuclear genome. {yields} Dilution of untreated genomic DNA leads to dilution bias. {yields} Unique primers and template pretreatment are needed to accurately measure mitochondrial DNA content. -- Abstract: Circulating mitochondrial DNA (MtDNA) is a potential non-invasive biomarker of cellular mitochondrial dysfunction, the latter known to be central to a wide range of human diseases. Changes in MtDNA are usually determined by quantification of MtDNA relative to nuclear DNA (Mt/N) using real time quantitative PCR. We propose that themore » methodology for measuring Mt/N needs to be improved and we have identified that current methods have at least one of the following three problems: (1) As much of the mitochondrial genome is duplicated in the nuclear genome, many commonly used MtDNA primers co-amplify homologous pseudogenes found in the nuclear genome; (2) use of regions from genes such as {beta}-actin and 18S rRNA which are repetitive and/or highly variable for qPCR of the nuclear genome leads to errors; and (3) the size difference of mitochondrial and nuclear genomes cause a 'dilution bias' when template DNA is diluted. We describe a PCR-based method using unique regions in the human mitochondrial genome not duplicated in the nuclear genome; unique single copy region in the nuclear genome and template treatment to remove dilution bias, to accurately quantify MtDNA from human samples.« less

Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents.

PubMed

Ter Laak, E A; Pijpers, A; Noordergraaf, J H; Schoevers, E C; Verheijden, J H

1991-02-01

The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar.

Comparison of methods for in vitro testing of susceptibility of porcine Mycoplasma species to antimicrobial agents.

PubMed Central

Ter Laak, E A; Pijpers, A; Noordergraaf, J H; Schoevers, E C; Verheijden, J H

1991-01-01

The MICs of 18 antimicrobial agents used against strains of three porcine Mycoplasma species were determined by a serial broth dilution method. Twenty field strains of M. hyorhinis, ten field strains of M. hyopneumoniae, six field strains of M. flocculare, and the type strains of these species were tested. Twelve field strains and the type strain of M. hyorhinis were also tested by an agar dilution method. Tests were read at various time points. When the broth dilution method was used, the final MIC had to be read 2 days after color changes had stopped. MICs of tetracycline, oxytetracycline, doxycycline, and minocycline were low for the three Mycoplasma species tested. MICs of chlortetracycline were 8 to 16 times higher than MICs of the other tetracyclines. Spiramycin, tylosin, kitasamycin, spectinomycin, tiamulin, lincomycin, and clindamycin were effective against all strains of M. hyorhinis and M. hyopneumoniae. The quinolones were highly effective against M. hyopneumoniae but less effective against M. hyorhinis. The susceptibility patterns for M. hyopneumoniae and M. flocculare were similar. PMID:2024954

Comparative techno-economic analysis of steam explosion, dilute sulfuric acid, ammonia fiber explosion and biological pretreatments of corn stover.

PubMed

Baral, Nawa Raj; Shah, Ajay

2017-05-01

Pretreatment is required to destroy recalcitrant structure of lignocelluloses and then transform into fermentable sugars. This study assessed techno-economics of steam explosion, dilute sulfuric acid, ammonia fiber explosion and biological pretreatments, and identified bottlenecks and operational targets for process improvement. Techno-economic models of these pretreatment processes for a cellulosic biorefinery of 113.5 million liters butanol per year excluding fermentation and wastewater treatment sections were developed using a modelling software-SuperPro Designer. Experimental data of the selected pretreatment processes based on corn stover were gathered from recent publications, and used for this analysis. Estimated sugar production costs ($/kg) via steam explosion, dilute sulfuric acid, ammonia fiber explosion and biological methods were 0.43, 0.42, 0.65 and 1.41, respectively. The results suggest steam explosion and sulfuric acid pretreatment methods might be good alternatives at present state of technology and other pretreatment methods require research and development efforts to be competitive with these pretreatment methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

Determination of Glyphosate, its Degradation Product Aminomethylphosphonic Acid, and Glufosinate, in Water by Isotope Dilution and Online Solid-Phase Extraction and Liquid Chromatography/Tandem Mass Spectrometry

USGS Publications Warehouse

Meyer, Michael T.; Loftin, Keith A.; Lee, Edward A.; Hinshaw, Gary H.; Dietze, Julie E.; Scribner, Elisabeth A.

2009-01-01

The U.S. Geological Survey method (0-2141-09) presented is approved for the determination of glyphosate, its degradation product aminomethylphosphonic acid (AMPA), and glufosinate in water. It was was validated to demonstrate the method detection levels (MDL), compare isotope dilution to standard addition, and evaluate method and compound stability. The original method USGS analytical method 0-2136-01 was developed using liquid chromatography/mass spectrometry and quantitation by standard addition. Lower method detection levels and increased specificity were achieved in the modified method, 0-2141-09, by using liquid chromatography/tandem mass spectrometry (LC/MS/MS). The use of isotope dilution for glyphosate and AMPA and pseudo isotope dilution of glufosinate in place of standard addition was evaluated. Stable-isotope labeled AMPA and glyphosate were used as the isotope dilution standards. In addition, the stability of glyphosate and AMPA was studied in raw filtered and derivatized water samples. The stable-isotope labeled glyphosate and AMPA standards were added to each water sample and the samples then derivatized with 9-fluorenylmethylchloroformate. After derivatization, samples were concentrated using automated online solid-phase extraction (SPE) followed by elution in-line with the LC mobile phase; the compounds separated and then were analyzed by LC/MS/MS using electrospray ionization in negative-ion mode with multiple-reaction monitoring. The deprotonated derivatized parent molecule and two daughter-ion transition pairs were identified and optimized for glyphosate, AMPA, glufosinate, and the glyphosate and AMPA stable-isotope labeled internal standards. Quantitative comparison between standard addition and isotope dilution was conducted using 473 samples analyzed between April 2004 and June 2006. The mean percent difference and relative standard deviation between the two quantitation methods was 7.6 plus or minus 6.30 (n = 179), AMPA 9.6 plus or minus 8.35 (n = 206), and glufosinate 9.3 plus or minus 9.16 (n = 16). The analytical variation of the method, comparison of quantitation by isotope dilution and multipoint linear regressed standard curves, and method detection levels were evaluated by analyzing six sets of distilled-water, groundwater, and surface-water samples spiked in duplicate at 0.0, 0.05, 0.10 and 0.50 microgram per liter and analyzed on 6 different days during 1 month. The grand means of the normalized concentration percentage recovery for glyphosate, AMPA, and glufosinate among all three matrices and spiked concentrations ranged from 99 to 114 plus or minus 2 to 7 percent of the expected spiked concentration. The grand mean of the percentage difference between concentrations calculated by standard addition and linear regressed multipoint standard curves ranged from 8 to 15 plus or minus 2 to 9 percent for the three compounds. The method reporting levels calculated from all the 0.05- microgram per liter spiked samples were 0.02 microgram per liter for all three compounds. Compound stability experiments were conducted on 10 samples derivatized four times for periods between 136 to 269 days. The glyphosate and AMPA concentrations remained relatively constant in samples held up to 136 days before derivatization. The half life of glyphosate varied from 169 to 223 days in the underivatized samples. Derivatized samples were analyzed the day after derivitization, and again 54 and 64 days after derivatization. The derivatized samples analyzed at days 52 and 64 were within 20 percent of the concentrations of the derivatized samples analyzed the day after derivatization.

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

DOEpatents

Turick, C.E.; Apel, W.W.

1997-10-28

A method of reducing the concentration of Cr(VI) in a liquid aqueous residue comprises the steps of providing anaerobic Cr(VI) reducing bacteria, mixing the liquid aqueous residue with a nutrient medium to form a mixture, and contacting the mixture with the anaerobic Cr(VI) reducing bacteria such that Cr(VI) is reduced to Cr(III). The anaerobic Cr(VI) reducing bacteria appear to be ubiquitous in soil and can be selected by collecting a soil sample, diluting the soil sample with a sterile diluent to form a diluted sample, mixing the diluted sample with an effective amount of a nutrient medium and an effective amount of Cr(VI) to form a mixture, and incubating the mixture in the substantial absence of oxygen such that growth of Cr(VI) sensitive microorganisms is inhibited and growth of the anaerobic Cr(VI) reducing bacteria is stimulated. A method of in situ bioremediation of Cr(VI) contaminated soil and/or groundwater is also disclosed. 10 figs.

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

DOEpatents

Turick, Charles E.; Apel, William W.

1997-10-28

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

Use of High-Resolution Continuum Source Flame Atomic Absorption Spectrometry (HR-CS FAAS) for Sequential Multi-Element Determination of Metals in Seawater and Wastewater Samples

NASA Astrophysics Data System (ADS)

Peña-Vázquez, E.; Barciela-Alonso, M. C.; Pita-Calvo, C.; Domínguez-González, R.; Bermejo-Barrera, P.

2015-09-01

The objective of this work is to develop a method for the determination of metals in saline matrices using high-resolution continuum source flame atomic absorption spectrometry (HR-CS FAAS). Module SFS 6 for sample injection was used in the manual mode, and flame operating conditions were selected. The main absorption lines were used for all the elements, and the number of selected analytical pixels were 5 (CP±2) for Cd, Cu, Fe, Ni, Pb and Zn, and 3 pixels for Mn (CP±1). Samples were acidified (0.5% (v/v) nitric acid), and the standard addition method was used for the sequential determination of the analytes in diluted samples (1:2). The method showed good precision (RSD(%) < 4%, except for Pb (6.5%)) and good recoveries. Accuracy was checked after the analysis of an SPS-WW2 wastewater reference material diluted with synthetic seawater (dilution 1:2), showing a good agreement between certified and experimental results.

Pulsed Dilution Method for the Recovery of Aggregated Mouse TNF-α.

PubMed

Mahmoodi, Merat; Ghodsi, Maryam; Moghadam, Malihe; Sankian, Mojtaba

2017-04-01

The expression of mouse tumor necrosis factor alpha (TNF-α) in Escherichia coli is a favorable way to get high yield of protein; however, the formation of cytoplasmic inclusion bodies, which is the consequence of insoluble accumulated proteins, is a major obstacle in this system. To overcome this obstacle, we used a pulsed dilution method to convert the product to its native conformation. Reducing agent and guanidine hydrochloride were used to solubilize inclusion bodies formed after TNF-(α) expression. Then, the refolding procedure was performed by pulsed dilution of the denatured protein into a refolding buffer. The properly-folded protein was purified by metal affinity chromatography. SDS-PAGE showed a 19.9 kDa band related to the mature TNF-(α) protein. The protein was recognized by anti-mouse TNF-(α) on western blots. The final concentration of the purified recombinant TNF-(α) was 62.5 µg/mL. Our study demonstrates the efficiency of this method to produce a high yield of folded mature TNF- (α).

Measurement of LNAPL flow using single-well tracer dilution techniques.

PubMed

Sale, Tom; Taylor, Geoffrey Ryan; Iltis, Gabriel; Lyverse, Mark

2007-01-01

This paper describes the use of single-well tracer dilution techniques to resolve the rate of light nonaqueous phase liquid (LNAPL) flow through wells and the adjacent geologic formation. Laboratory studies are presented in which a fluorescing tracer is added to LNAPL in wells. An in-well mixer keeps the tracer well mixed in the LNAPL. Tracer concentrations in LNAPL are measured through time using a fiber optic cable and a spectrometer. Results indicate that the rate of tracer depletion is proportional to the rate of LNAPL flow through the well and the adjacent formation. Tracer dilution methods are demonstrated for vertically averaged LNAPL Darcy velocities of 0.00048 to 0.11 m/d and LNAPL thicknesses of 9 to 24 cm. Over the range of conditions studied, results agree closely with steady-state LNAPL flow rates imposed by pumping. A key parameter for estimating LNAPL flow rates in the formation is the flow convergence factor alpha. Measured convergence factors for 0.030-inch wire wrap, 0.030-inch-slotted polyvinyl chloride (PVC), and 0.010-inch-slotted PVC are 1.7, 0.91, and 0.79, respectively. In addition, methods for using tracer dilution data to determine formation transmissivity to LNAPL are presented. Results suggest that single-well tracer dilution techniques are a viable approach for measuring in situ LNAPL flow and formation transmissivity to LNAPL.

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

Beavin, P. Jr.

A previously published method for determining zirconium in antiperspirant aerosols was collaboratively studied by 7 laboratories. The method consists of 2 procedures: a rapid dilution procedure for soluble zirconium compounds or a lengthier fusion procedure for total zirconium followed by colorimetric determination. The collaborators were asked to perform the following: Spiking materials representing 4 levels of soluble zirconium were added to weighed portions of a zirconium-free cream base concentrate and the portions were assayed by the dilution procedure. Spiking materials representing 4 levels of zirconium in either the soluble or the insoluble form (or as a mixture) were also addedmore » to portions of the same concentrate and these portions were assayed by the fusion procedure. They were also asked to concentrate and assay, by both procedures, 2 cans each of 2 commercial aerosol antiperspirants containing zirconyl hydroxychloride. The average percent recoveries and standard deviations for spiked samples were 99.8-100.2 and 1.69-2.71, respectively, for soluble compounds determined by the dilution procedure, and 93.8-97.4 and 3.09-4.78, respectively, for soluble and/or insoluble compounds determined by the fusion procedure. The average perent zirconium found by the dilution procedure in the 2 commercial aerosol products was 0.751 and 0.792. Insufficient collaborative results were received for the fusion procedure for statistical evaluation. The dilution procedure has been adopted as official first action.« less

Development of reproducible assays for polygalacturonase and pectinase.

PubMed

Li, Qian; Coffman, Anthony M; Ju, Lu-Kwang

2015-05-01

Polygalacturonase and pectinase activities reported in the literature were measured by several different procedures. These procedures do not give comparable results, partly owing to the complexity of the substrates involved. This work was aimed at developing consistent and efficient assays for polygalacturonase and pectinase activities, using polygalacturonic acid and citrus pectin, respectively, as the substrate. Different enzyme mixtures produced by Aspergillus niger and Trichoderma reesei with different inducing carbon sources were used for the method development. A series of experiments were conducted to evaluate the incubation time, substrate concentration, and enzyme dilution. Accordingly, for both assays the recommended (optimal) hydrolysis time is 30min and substrate concentration is 5g/L. For polygalacturonase, the sample should be adjusted to have 0.3-0.8U/mL polygalacturonase activity, because in this range the assay outcomes were consistent (independent of dilution factors). Such a range did not exist for the pectinase assay. The recommended procedure is to assay the sample at multiple (at least 2) dilution factors and determine, by linear interpolation, the dilution factor that would release reducing sugar equivalent to 0.4g/L d-galacturonic acid, and then calculate the activity of the sample accordingly (dilution factor×0.687U/mL). Validation experiments showed consistent results using these assays. Effects of substrate preparation methods were also examined. Copyright © 2015 Elsevier Inc. All rights reserved.

Method of determining the optimal dilution ratio for fluorescence fingerprint of food constituents.

PubMed

Trivittayasil, Vipavee; Tsuta, Mizuki; Kokawa, Mito; Yoshimura, Masatoshi; Sugiyama, Junichi; Fujita, Kaori; Shibata, Mario

2015-01-01

Quantitative determination by fluorescence spectroscopy is possible because of the linear relationship between the intensity of emitted fluorescence and the fluorophore concentration. However, concentration quenching may cause the relationship to become nonlinear, and thus, the optimal dilution ratio has to be determined. In the case of fluorescence fingerprint (FF) measurement, fluorescence is measured under multiple wavelength conditions and a method of determining the optimal dilution ratio for multivariate data such as FFs has not been reported. In this study, the FFs of mixed solutions of tryptophan and epicatechin of different concentrations and composition ratios were measured. Principal component analysis was applied, and the resulting loading plots were found to contain useful information about each constituent. The optimal concentration ranges could be determined by identifying the linear region of the PC score plotted against total concentration.

Antibacterial activity of Tribulus terrestris and its synergistic effect with Capsella bursa-pastoris and Glycyrrhiza glabra against oral pathogens: an in-vitro study

PubMed Central

Soleimanpour, Saman; Sedighinia, Fereshteh Sadat; Safipour Afshar, Akbar; Zarif, Reza; Ghazvini, Kiarash

2015-01-01

Objective: In this study, antimicrobial activities of an ethanol extract of Tribulus terrestris aloneand in combination with Capsella bursa-pastoris and Glycyrrhiza glabra were examined in vitro against six pathogens namely Streptococcus mutans, Streptococcus sanguis, Actinomyces viscosus, Enterococcus faecalis Staphylococcus aureus, and Escherichia coli. Materials and methods: Antibacterial activities of the extracts were examined using disc and well diffusion methods and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ethanol extracts were determined against these microorganisms using agar and broth dilution methods. Chlorhexidine was used as positive control. Results: Tribulus terrestris extract exhibited good antibacterial activity against all bacteria. Antibacterial activity of mixed extract was evaluated and exhibited that mixed extract was more effective against all bacteria than any of the cases alone which indicates the synergistic effect between these three extracts (p˂0.05). No strain showed resistance against these extracts. In agar dilution, Tribulus terrestris exhibited MIC values ranging from 35.0 to 20.0 mg/ml and mixed extract showed MIC values ranging from 12.5 to 5.0 mg/ml. The results of broth dilution method were consistent with the findings of the agar dilution method. Conclusion: This in-vitro study was a preliminary evaluation of antibacterial activity of the plants. It provided scientific evidence to support uses of T. terrestris and its mixture with C. bursa-pastoris and G. glabra for the treatment of oral infections. In-vivo studies are also required to better evaluate the effect of these extracts. PMID:26101754

Diluted magnetic semiconductor nanowires exhibiting magnetoresistance

DOEpatents

Yang, Peidong [El Cerrito, CA; Choi, Heonjin [Seoul, KR; Lee, Sangkwon [Daejeon, KR; He, Rongrui [Albany, CA; Zhang, Yanfeng [El Cerrito, CA; Kuykendal, Tevye [Berkeley, CA; Pauzauskie, Peter [Berkeley, CA

2011-08-23

A method for is disclosed for fabricating diluted magnetic semiconductor (DMS) nanowires by providing a catalyst-coated substrate and subjecting at least a portion of the substrate to a semiconductor, and dopant via chloride-based vapor transport to synthesize the nanowires. Using this novel chloride-based chemical vapor transport process, single crystalline diluted magnetic semiconductor nanowires Ga.sub.1-xMn.sub.xN (x=0.07) were synthesized. The nanowires, which have diameters of .about.10 nm to 100 nm and lengths of up to tens of micrometers, show ferromagnetism with Curie temperature above room temperature, and magnetoresistance up to 250 Kelvin.

Oxalate Content of Different Drinkable Dilutions of Tea Infusions after Different Brewing Times

PubMed Central

Lotfi Yagin, Neda; Mahdavi, Reza; Nikniaz, Zeinab

2012-01-01

Background: The aims of this study were to determine the effect of different brewing times and diluting on oxalate content of loose-packed black teas consumed in Tabriz, Iran. Methods: The oxalate content of black teas after brewing for 5, 10, 15, 30, 60 minutes was measured in triplicate by enzymatic assay. In order to attain the most acceptable dilution of tea infusions, tea samples which were brewed for 15, 30 and 60 minutes were diluted two (120 ml), three (80 ml) and four (60 ml) times respectively. Results: There was a stepwise increase in oxalate concentrations associated with increased brewing times (P< 0.001) with oxalate contents ranging from 4.4 mg/240 ml for the 5 min to 6.3 mg/240 ml for 60 min brewing times, respectively. There were significant differences between the mean oxalate content of different dilutions after brewing for 15, 30 and 60 minutes (P< 0.001). Conclusion: The oxalate content of Iranian consumed black tea after different brewing times and different dilution was below the recommended levels. Therefore, it seems that consumption of black tea several times per day would not pose significant health risk in kidney stone patients and susceptible individuals. PMID:24688937

[Comparison of single-indicator thermodilution versus gravimetric measurement in determination of extra-vascular lung water in dogs with acute respiratory distress syndrome].

PubMed

Shen, Ju-fang; Qiu, Hai-bo; Yang, Yi; Liu, Song-qiao; Chen, Yong-ming; Li, Jia-qiong; Wu, Bin; Ding, Hui-min

2006-06-01

To compare the measurement of extra-vascular lung water (EVLW) by a single-indicator dilution technique and measurement obtained by gravimetry in different types of acute respiratory distress syndrome (ARDS). Thirty-three dogs were randomly assigned to three groups: control group, oleic acid group and hydrochloric acid group. ARDS was reproduced by either intravenous injection of oleic acid or intratracheal instillation of hydrochloric acid. EVLW was measured before ARDS, at the onset of ARDS and 10 hours after ARDS by a single indicator dilution technique. Ten hours after ARDS, dogs were sacrificed and then EVLW was quantitated by a gravimetric measurement (golden standard). Hemodynamics and pulmonary gas exchange were determined. There was a close positive correlation (r=0.8820, P<0.05) between single indicator dilution and gravimetric measurements. However, the measurement with the single indicator dilution was consistently higher than the gravimetric measurement. In the control group, there was a positive correlation (r=0.9870, P<0.05) between the values of EVLW as measured by single indicator dilution and by gravimetric measurements. In the oleic acid group, there was also a significant correlation (r=0.9360, P<0.05) between the values of EVLW as measured by single indicator dilution and by gravimetric measurements. In the hydrochloric acid group, correlation (r=0.7950, P<0.05) was also found between EVLW as measured by the two methods. However, the correlation found was lower in the hydrochloric acid group than those in other two groups. Hydrochloric acid instillation resulted in a significant increase in shunting and the partial pressure of carbon dioxide in artery (PaCO(2)) compared with oleic acid group at 10 hours after ARDS. The results of measuring EVLW using single indicator dilution measurement are closely related with those of gravimetric measurement in ARDS, however, the correlations varies with the methods of reproduction of ARDS.

Reproducibility of CSF quantitative culture methods for estimating rate of clearance in cryptococcal meningitis.

PubMed

Dyal, Jonathan; Akampurira, Andrew; Rhein, Joshua; Morawski, Bozena M; Kiggundu, Reuben; Nabeta, Henry W; Musubire, Abdu K; Bahr, Nathan C; Williams, Darlisha A; Bicanic, Tihana; Larsen, Robert A; Meya, David B; Boulware, David R

2016-05-01

Quantitative cerebrospinal fluid (CSF) cultures provide a measure of disease severity in cryptococcal meningitis. The fungal clearance rate by quantitative cultures has become a primary endpoint for phase II clinical trials. This study determined the inter-assay accuracy of three different quantitative culture methodologies. Among 91 participants with meningitis symptoms in Kampala, Uganda, during August-November 2013, 305 CSF samples were prospectively collected from patients at multiple time points during treatment. Samples were simultaneously cultured by three methods: (1) St. George's 100 mcl input volume of CSF with five 1:10 serial dilutions, (2) AIDS Clinical Trials Group (ACTG) method using 1000, 100, 10 mcl input volumes, and two 1:100 dilutions with 100 and 10 mcl input volume per dilution on seven agar plates; and (3) 10 mcl calibrated loop of undiluted and 1:100 diluted CSF (loop). Quantitative culture values did not statistically differ between St. George-ACTG methods (P= .09) but did for St. George-10 mcl loop (P< .001). Repeated measures pairwise correlation between any of the methods was high (r≥0.88). For detecting sterility, the ACTG-method had the highest negative predictive value of 97% (91% St. George, 60% loop), but the ACTG-method had occasional (∼10%) difficulties in quantification due to colony clumping. For CSF clearance rate, St. George-ACTG methods did not differ overall (mean -0.05 ± 0.07 log10CFU/ml/day;P= .14) on a group level; however, individual-level clearance varied. The St. George and ACTG quantitative CSF culture methods produced comparable but not identical results. Quantitative cultures can inform treatment management strategies. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Sensitivity of the activated partial thromboplastin time, the dilute Russell's viper venom time, and the kaolin clotting time for the detection of the lupus anticoagulant: a direct comparison using plasma dilutions.

PubMed

Martin, B A; Branch, D W; Rodgers, G M

1996-01-01

Increasing dilutions of lupus anticoagulant (LA) plasmas from twelve patients were used to directly compare the sensitivity of four tests for LA. The tests evaluated were the modified Bell and Alton activated partial thromboplastin time (APTT), an APTT using a commercially prepared partial thromboplastin (Platelin LS APTT), a modified dilute Russell's viper venom time (DRVVT), and a modified kaolin clotting time (KCT). LAs were detected in all twelve plasmas by each of three tests and eleven of twelve plasmas in a fourth test when undiluted patient plasma was used. Repeating the tests after diluting the LA plasmas with normal platelet-free plasma (PFP) showed that the KCT was the most sensitive test for LA, detecting eleven of twelve LAs at a dilution of 10% patient plasma and ten of twelve LAs at a dilution of 5% patient plasma. The modified Bell and Alton APTT and the modified DRVVT had similar sensitivities at a patient plasma concentration of 10%, detecting seven of twelve and eight of twelve LAs, respectively. The Platelin LS APTT detected only four of twelve LAs at a patient plasma concentration of 10%. Our results indicate that the modified KCT is a sensitive method for the detection of LAs. The modified Bell and Alton APTT and the DRVVT were less sensitive.

Gluconeogenesis from labeled carbon: estimating isotope dilution

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

Kelleher, J.K.

1986-03-01

To estimate the rate of gluconeogenesis from steady-state incorporation of labeled 3-carbon precursors into glucose, isotope dilution must be considered so that the rate of labeling of glucose can be quantitatively converted to the rate of gluconeogenesis. An expression for the value of this isotope dilution can be derived using mathematical techniques and a model of the tricarboxylic acid (TCA) cycle. The present investigation employs a more complex model than that used in previous studies. This model includes the following pathways that may affect the correction for isotope dilution: 1) flux of 3-carbon precursor to the oxaloacetate pool via acetyl-CoAmore » and the TCA cycle; 2) flux of 4- or 5-carbon compounds into the TCA cycle; 3) reversible flux between oxaloacetate (OAA) and pyruvate and between OAA and fumarate; 4) incomplete equilibrium between OAA pools; and 5) isotope dilution of 3-carbon tracers between the experimentally measured pool and the precursor for the TCA-cycle OAA pool. Experimental tests are outlined which investigators can use to determine whether these pathways are significant in a specific steady-state system. The study indicated that flux through these five pathways can significantly affect the correction for isotope dilution. To correct for the effects of these pathways an alternative method for calculating isotope dilution is proposed using citrate to relate the specific activities of acetyl-CoA and OAA.« less

Sperm cryopreservation of African catfish, Clarias gariepinus: cryoprotectants, freezing rates and sperm:egg dilution ratio.

PubMed

Viveiros, A T; So, N; Komen, J

2000-12-01

Methods for cryopreserving spermatozoa and optimizing sperm:egg dilution ratio in African catfish Clarias gariepinus were developed. Five percent to 25% DMSO and methanol were tested as cryoprotectants, by diluting semen in Ginzburg fish ringer and freezing in 1-milliliter cryovials in a programmable freezer. To avoid an excess of spermatozoa per egg, post-thaw semen was diluted 1:20, 1:200 or 1:2,000 before fertilization. Highest hatching rates were obtained by spermatozoa frozen in 10% methanol and post-thaw diluted to 1:200. Then, slow freezing rates (-2, -5 or -10 degrees C/min) to various endpoint temperatures (range -25 to -70 degrees C) before fast freezing in liquid nitrogen (LN2) were evaluated. Hatching rates equal to control (P > 0.05) were obtained by spermatozoa frozen at -5 degrees C/min to -45 to -50 degrees C and at -10 degrees C/min to -55 degrees C. In 3-step freezing programs, at -5 degrees C/min, the effect of holding spermatozoa for 0, 2 or 5 min at -30, -35 or -40 degrees C before fast freezing in LN2 was analyzed. Hatching rates equal to control (P > 0.05) were produced by spermatozoa frozen to, and held at, -35 degrees C for 5 min and at -40 degrees C for 2 or 5 min. Finally, frozen spermatozoa (10% methanol, -5 degrees C/min, 5-min hold at -40 degrees C, LN2, post-thaw diluted to 1:200) were tested in on-farm fertilization conditions. Again, no difference (P > 0.05) in hatching rate was observed between frozen and fresh spermatozoa. Cryopreservation offers utility as a routine method of sperm storage and management for catfish.

Changes of jugular venous blood temperature associated with measurements of cerebral blood flow using the transcerebral double-indicator dilution technique.

PubMed

Mielck, F; Bräuer, A; Radke, O; Hanekop, G; Loesch, S; Friedrich, M; Hilgers, R; Sonntag, H

2004-04-01

The transcerebral double-indicator dilution technique is a recently developed method to measure global cerebral blood flow at bedside. It is based on bolus injection of ice-cold indocyanine green dye and simultaneous recording of resulting thermo- and dye-dilution curves in the aorta and the jugular bulb. However, with this method 40 mL of ice-cold solution is administered as a bolus. Therefore, this prospective clinical study was performed to elucidate the effects of repeated administration of indicator on absolute blood temperature and on cerebral blood flow and metabolism. The investigation was performed in nine male patients scheduled for elective coronary artery bypass grafting. Absolute blood temperature was measured in the jugular bulb and in the aorta before and after repeated measurements using the transcerebral double-indicator dilution technique. During the investigated time course, the blood temperature in the jugular bulb, compared to the aorta, was significantly higher with a mean difference of 0.21 degrees C. The administration of an ice-cold bolus reduced the mean blood temperature by 0.06 degrees C in the jugular bulb as well as in the aorta. After the transcerebral double-indicator dilution measurements a temperature recovery to baseline conditions was not observed during the investigated time period. Cerebral blood flow and cerebral metabolism did not change during the investigated time period. Repeated measurements with the transcerebral double-indicator dilution technique do not affect absolute jugular bulb blood temperatures negatively. Global cerebral blood flow and metabolism measurements remain unaltered. However, accuracy and resolution of this technique is not high enough to detect the effect of minor changes of physiological variables.

Assessment of body fat in the pony: part II. Validation of the deuterium oxide dilution technique for the measurement of body fat.

PubMed

Dugdale, A H A; Curtis, G C; Milne, E; Harris, P A; Argo, C Mc

2011-09-01

Excessive accumulations or depletions of body fat have been associated with increased morbidity and mortality in horses and ponies. An objective, minimally-invasive method to accurately quantify body fat in living animals is required to aid nutritional management and define welfare/performance limits. To compare deuterium oxide (D(2) O) dilution-derived estimates of total body water (TBW) and body fat with values obtained by 'gold standard' proximate analysis and cadaver dissection. D(2) O dilution offers a valid method for the determination of TBW and body fat in equids. Seven mature (mean ± s.e. 13 ± 3 years, 212 ± 14 kg, body condition scores 1.25-7/9), healthy, Welsh Mountain pony mares, destined for euthanasia (for nonresearch purposes) were used. Blood samples were collected before and 4 h after D(2) O (0.11-0.13 g/kg bwt, 99.8 atom percent excess) administration. Plasma was analysed by gas isotope ratio mass spectrometry following filtration and zinc reduction. After euthanasia, white adipose tissue (WAT) mass was recorded before all body tissues were analysed by proximate chemical analyses. D(2) O-derived estimates of TBW and body fat were strongly associated with proximate analysis- and dissection-derived values (all r(2) >0.97, P≤0.0001). Bland-Altman analyses demonstrated good agreements between methods. D(2) O dilution slightly overestimated TBW (0.79%, limits of agreement (LoA) -3.75-2.17%) and underestimated total body lipid (1.78%, LoA -0.59-4.15%) and dissected WAT (0.72%, LoA -2.77-4.21%). This study provides the first validation of the D(2) O dilution method for the minimally-invasive, accurate, repeatable and objective measurement of body water and fat in living equids. © 2011 EVJ Ltd.

Regression dilution bias: tools for correction methods and sample size calculation.

PubMed

Berglund, Lars

2012-08-01

Random errors in measurement of a risk factor will introduce downward bias of an estimated association to a disease or a disease marker. This phenomenon is called regression dilution bias. A bias correction may be made with data from a validity study or a reliability study. In this article we give a non-technical description of designs of reliability studies with emphasis on selection of individuals for a repeated measurement, assumptions of measurement error models, and correction methods for the slope in a simple linear regression model where the dependent variable is a continuous variable. Also, we describe situations where correction for regression dilution bias is not appropriate. The methods are illustrated with the association between insulin sensitivity measured with the euglycaemic insulin clamp technique and fasting insulin, where measurement of the latter variable carries noticeable random error. We provide software tools for estimation of a corrected slope in a simple linear regression model assuming data for a continuous dependent variable and a continuous risk factor from a main study and an additional measurement of the risk factor in a reliability study. Also, we supply programs for estimation of the number of individuals needed in the reliability study and for choice of its design. Our conclusion is that correction for regression dilution bias is seldom applied in epidemiological studies. This may cause important effects of risk factors with large measurement errors to be neglected.

Development and Implementation of a Coagulation Factor Testing Method Utilizing Autoverification in a High-volume Clinical Reference Laboratory Environment

PubMed Central

Riley, Paul W.; Gallea, Benoit; Valcour, Andre

2017-01-01

Background: Testing coagulation factor activities requires that multiple dilutions be assayed and analyzed to produce a single result. The slope of the line created by plotting measured factor concentration against sample dilution is evaluated to discern the presence of inhibitors giving rise to nonparallelism. Moreover, samples producing results on initial dilution falling outside the analytic measurement range of the assay must be tested at additional dilutions to produce reportable results. Methods: The complexity of this process has motivated a large clinical reference laboratory to develop advanced computer algorithms with automated reflex testing rules to complete coagulation factor analysis. A method was developed for autoverification of coagulation factor activity using expert rules developed with on an off the shelf commercially available data manager system integrated into an automated coagulation platform. Results: Here, we present an approach allowing for the autoverification and reporting of factor activity results with greatly diminished technologist effort. Conclusions: To the best of our knowledge, this is the first report of its kind providing a detailed procedure for implementation of autoverification expert rules as applied to coagulation factor activity testing. Advantages of this system include ease of training for new operators, minimization of technologist time spent, reduction of staff fatigue, minimization of unnecessary reflex tests, optimization of turnaround time, and assurance of the consistency of the testing and reporting process. PMID:28706751

Determination of (187)Os in molybdenite by ICP-MS with neutron-induced (186)Os and (188)Os spikes.

PubMed

Qu, W; Du, A; Zhao, D

2001-10-31

The article describes a method for the determination of (187)Os in molybdenite by isotope dilution inductively coupled plasma-mass spectrometry (ID-ICP-MS) with neutron-induced (186)Os and (188)Os spike. The spike used in the present work was prepared in line with the principle by which artificial nuclides are produced in a nuclear reaction. The concentration and isotopic composition of osmium in the prepared spike were evaluated accurately with the isotope dilution method, using negative thermal ion mass spectrometry (N-TIMS). The advantage of this method is that using (186)Os and (188)Os double spikes can effectively compensate for the mass discrimination effects of ICP-MS. Thus, the common correction practice for mass bias in the isotope dilution method with a single spike is unnecessary. In addition, the method enables one to reduce the determined error arising from instrumental instability. The precision for the (187)Os/((186)Os+(188)Os) ratio was approximately 2% (2sigma, RSD), but in the case of (187)Os/(186)Os, (187)Os/(188)Os and (186)Os/(188)Os, precision ranged from 2.0 to 8% (2sigma, RSD). The results for (187)Os concentration in a molybdenite sample determined with this method showed good agreement with reference values.

Trace determination of zinc by substoichiometric isotope dilution analysis

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

Sandhya, D.; Priya, S.; Subramanian, M.O.S.

1996-09-01

A radiometric method based on substoichiometric isotope dilution analysis using 1,10-phenanthroline and a substoichiometric amount of eosin was developed for determining trace amounts of zinc. Evaluation of various metal ion interferences shows that as little as 0.2 {mu}g Zn could be determined in an aqueous-phase volume of 60 mL. The method has been successfully applied to the determination of Zn in city waste incineration ash, cadmium metal, Fourts-B tablets, Boro-plus ointment, and magnesium alloy samples. 12 refs., 3 figs., 3 tabs.

40 CFR Appendix B to Part 80 - Test Methods for Lead in Gasoline

Code of Federal Regulations, 2014 CFR

2014-07-01

... one liter. 4.6 Iodine Solution—Dissolve and dilute 3.0 g iodine crystals with Toluene to 100 ml. 4.7...): Dissolve and dilute 3.0 g iodine crystals (American Chemical Society) with toluene (Burdick & Jackson Lab... and aligned according to Section 7 and the instrument has had substantial time for warm-up. 8.2Auto...

40 CFR Appendix B to Part 80 - Test Methods for Lead in Gasoline

Code of Federal Regulations, 2010 CFR

2010-07-01

... one liter. 4.6 Iodine Solution—Dissolve and dilute 3.0 g iodine crystals with Toluene to 100 ml. 4.7...): Dissolve and dilute 3.0 g iodine crystals (American Chemical Society) with toluene (Burdick & Jackson Lab... and aligned according to Section 7 and the instrument has had substantial time for warm-up. 8.2Auto...

40 CFR Appendix B to Part 80 - Test Methods for Lead in Gasoline

Code of Federal Regulations, 2013 CFR

2013-07-01

... one liter. 4.6 Iodine Solution—Dissolve and dilute 3.0 g iodine crystals with Toluene to 100 ml. 4.7...): Dissolve and dilute 3.0 g iodine crystals (American Chemical Society) with toluene (Burdick & Jackson Lab... and aligned according to Section 7 and the instrument has had substantial time for warm-up. 8.2Auto...

40 CFR Appendix B to Part 80 - Test Methods for Lead in Gasoline

Code of Federal Regulations, 2011 CFR

2011-07-01

... one liter. 4.6 Iodine Solution—Dissolve and dilute 3.0 g iodine crystals with Toluene to 100 ml. 4.7...): Dissolve and dilute 3.0 g iodine crystals (American Chemical Society) with toluene (Burdick & Jackson Lab... and aligned according to Section 7 and the instrument has had substantial time for warm-up. 8.2Auto...

40 CFR Appendix B to Part 80 - Test Methods for Lead in Gasoline

Code of Federal Regulations, 2012 CFR

2012-07-01

... one liter. 4.6 Iodine Solution—Dissolve and dilute 3.0 g iodine crystals with Toluene to 100 ml. 4.7...): Dissolve and dilute 3.0 g iodine crystals (American Chemical Society) with toluene (Burdick & Jackson Lab... and aligned according to Section 7 and the instrument has had substantial time for warm-up. 8.2Auto...

Modeling mass transfer and reaction of dilute solutes in a ternary phase system by the lattice Boltzmann method

NASA Astrophysics Data System (ADS)

Fu, Yu-Hang; Bai, Lin; Luo, Kai-Hong; Jin, Yong; Cheng, Yi

2017-04-01

In this work, we propose a general approach for modeling mass transfer and reaction of dilute solute(s) in incompressible three-phase flows by introducing a collision operator in lattice Boltzmann (LB) method. An LB equation was used to simulate the solute dynamics among three different fluids, in which the newly expanded collision operator was used to depict the interface behavior of dilute solute(s). The multiscale analysis showed that the presented model can recover the macroscopic transport equations derived from the Maxwell-Stefan equation for dilute solutes in three-phase systems. Compared with the analytical equation of state of solute and dynamic behavior, these results are proven to constitute a generalized framework to simulate solute distributions in three-phase flows, including compound soluble in one phase, compound adsorbed on single-interface, compound in two phases, and solute soluble in three phases. Moreover, numerical simulations of benchmark cases, such as phase decomposition, multilayered planar interfaces, and liquid lens, were performed to test the stability and efficiency of the model. Finally, the multiphase mass transfer and reaction in Janus droplet transport in a straight microchannel were well reproduced.

Optimization and validation of enzyme-linked immunosorbent assay for the determination of endosulfan residues in food samples.

PubMed

Zhang, Yan; Liu, Jun W; Zheng, Wen J; Wang, Lei; Zhang, Hong Y; Fang, Guo Z; Wang, Shuo

2008-02-01

In this study, an enzyme-linked immunosorbent assay (ELISA) was optimized and applied to the determination of endosulfan residues in 20 different kinds of food commodities including vegetables, dry fruits, tea and meat. The limit of detection (IC(15)) was 0.8 microg kg(-1) and the sensitivity (IC(50)) was 5.3 microg kg(-1). Three simple extraction methods were developed, including shaking on the rotary shaker at 250 r min(-1) overnight, shaking on the rotary shaker for 1 h and thoroughly mixing for 2 min. Methanol was used as the extraction solvent in this study. The extracts were diluted in 0.5% fish skin gelatin (FG) in phosphate-buffered saline (PBS) at various dilutions in order to remove the matrix interference. For cabbage (purple and green), asparagus, Japanese green, Chinese cabbage, scallion, garland chrysanthemum, spinach and garlic, the extracts were diluted 10-fold; for carrots and tea, the extracts were diluted 15-fold and 900-fold, respectively. The extracts of celery, adzuki beans and chestnuts, were diluted 20-fold to avoid the matrix interference; ginger, vegetable soybean and peanut extracts were diluted 100-fold; mutton and chicken extracts were diluted 10-fold and for eel, the dilution was 40-fold. Average recoveries were 63.13-125.61%. Validation was conducted by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The results of this study will be useful to the wide application of an ELISA for the rapid determination of pesticides in food samples.

Stability of florfenicol in drinking water.

PubMed

Hayes, John M; Eichman, Jonathan; Katz, Terry; Gilewicz, Rosalia

2003-01-01

Florfenicol, a broad-spectrum antibiotic, is being developed for veterinary application as an oral concentrate intended for dilution with drinking water. When a drug product is dosed via drinking water in a farm setting, a number of variables, including pH, chlorine content, hardness of the water used for dilution, and container material, may affect its stability, leading to a decrease in drug potency. The stability of florfenicol after dilution of Florfenicol Drinking Water Concentrate Oral Solution, 23 mg/mL, with drinking water was studied. A stability-indicating, validated liquid chromatographic method was used to evaluate florfenicol stability at 25 degrees C at 5, 10, and 24 h after dilution. The results indicate that florfenicol is stable under a range of simulated field conditions, including various pipe materials and conditions of hard or soft and chlorinated or nonchlorinated water at low or high pH. Significant degradation (> 10%) was observed only for isolated combinations in galvanized pipes. Analysis indicated that the florfenicol concentration in 8 of the 12 water samples stored in galvanized pipes remained above 90% of the initial concentration (100 mg/L) for 24 h after dilution.

Dilution of boar ejaculates with BTS containing HEPES in place of bicarbonate immediately after ejaculation can reduce the increased inducibility of the acrosome reaction by treatment with calcium and calcium ionophore A23187, which is potentially associated with boar subfertility.

PubMed

Murase, Tetsuma; Imaeda, Noriaki; Yamada, Hiroto; Takasu, Masaki; Taguchi, Kazuo; Katoh, Tsutomu

2010-06-01

The present study investigated whether substitution of HEPES for bicarbonate in BTS (BTS-H) used to dilute boar ejaculates immediately after ejaculation could reduce the increased inducibility of the acrosome reaction by calcium and calcium ionophore A23187. When an ejaculate was split, diluted 5-fold with regular BTS (BTS-B) and BTS-H and stored at 17 C for 12 h or 60 h, the extender or storage time had no significant influence on sperm motility or viability measured by the eosin-nigrosin method. When spermatozoa diluted serially with BTS-B and stored (36 h) were stimulated with Ca2+ (3 mM) and A23187 (0.3 microM), the proportion of spermatozoa that underwent the acrosome reaction (% acrosome reactions) significantly increased as the magnifications of dilution increased (bicarbonate content almost unchanged by dilution). By contrast, the % acrosome reactions in spermatozoa similarly diluted and stored with BTS-H decreased with the increasing magnifications of dilution (bicarbonate decreased). Sperm motility immediately after the end of incubation without A23178 tended to be lower for BTS-H than BTS-B, and the ejaculates for BTS-H had a tendency to have a lower total protein in seminal plasma than those for BTS-B. These results implied that the samples for BTS-H could be used as a model for ejaculates possibly collected during summer and showing subfertility. When an ejaculate was split, diluted serially with BTS-B and BTS-H and stored, viability measured by staining with propidium iodide was extremely similar between the 2 extenders and among the different dilution magnifications, regardless of whether spermatozoa were washed (stored for 36-66 h) or not (stored for 66-72 h). These results suggest that boar ejaculate can be stored with BTS-H at least according to the results for sperm motility and viability and that hypersensitivity of spermatozoa to Ca2+ and A23187 potentially associated with boar subfertility could be lessened by diluting ejaculates with BTS-H.

Antibacterial activity of Tribulus terrestris and its synergistic effect with Capsella bursa-pastoris and Glycyrrhiza glabra against oral pathogens: an in-vitro study.

PubMed

Soleimanpour, Saman; Sedighinia, Fereshteh Sadat; Safipour Afshar, Akbar; Zarif, Reza; Ghazvini, Kiarash

2015-01-01

In this study, antimicrobial activities of an ethanol extract of Tribulus terrestris aloneand in combination with Capsella bursa-pastoris and Glycyrrhiza glabra were examined in vitro against six pathogens namely Streptococcus mutans, Streptococcus sanguis, Actinomyces viscosus, Enterococcus faecalis Staphylococcus aureus, and Escherichia coli. Antibacterial activities of the extracts were examined using disc and well diffusion methods and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ethanol extracts were determined against these microorganisms using agar and broth dilution methods. Chlorhexidine was used as positive control. Tribulus terrestris extract exhibited good antibacterial activity against all bacteria. Antibacterial activity of mixed extract was evaluated and exhibited that mixed extract was more effective against all bacteria than any of the cases alone which indicates the synergistic effect between these three extracts (p˂0.05). No strain showed resistance against these extracts. In agar dilution, Tribulus terrestris exhibited MIC values ranging from 35.0 to 20.0 mg/ml and mixed extract showed MIC values ranging from 12.5 to 5.0 mg/ml. The results of broth dilution method were consistent with the findings of the agar dilution method. This in-vitro study was a preliminary evaluation of antibacterial activity of the plants. It provided scientific evidence to support uses of T. terrestris and its mixture with C. bursa-pastoris and G. glabra for the treatment of oral infections. In-vivo studies are also required to better evaluate the effect of these extracts.

Short-term landfill methane emissions dependency on wind.

PubMed

Delkash, Madjid; Zhou, Bowen; Han, Byunghyun; Chow, Fotini K; Rella, Chris W; Imhoff, Paul T

2016-09-01

Short-term (2-10h) variations of whole-landfill methane emissions have been observed in recent field studies using the tracer dilution method for emissions measurement. To investigate the cause of these variations, the tracer dilution method is applied using 1-min emissions measurements at Sandtown Landfill (Delaware, USA) for a 2-h measurement period. An atmospheric dispersion model is developed for this field test site, which is the first application of such modeling to evaluate atmospheric effects on gas plume transport from landfills. The model is used to examine three possible causes of observed temporal emissions variability: temporal variability of surface wind speed affecting whole landfill emissions, spatial variability of emissions due to local wind speed variations, and misaligned tracer gas release and methane emissions locations. At this site, atmospheric modeling indicates that variation in tracer dilution method emissions measurements may be caused by whole-landfill emissions variation with wind speed. Field data collected over the time period of the atmospheric model simulations corroborate this result: methane emissions are correlated with wind speed on the landfill surface with R(2)=0.51 for data 2.5m above ground, or R(2)=0.55 using data 85m above ground, with emissions increasing by up to a factor of 2 for an approximately 30% increase in wind speed. Although the atmospheric modeling and field test are conducted at a single landfill, the results suggest that wind-induced emissions may affect tracer dilution method emissions measurements at other landfills. Copyright © 2016 Elsevier Ltd. All rights reserved.

Rapid Identification of Vancomycin Resistant Enterococcus Faecalis Clinical Isolates using a Sugar Fermentation Method

PubMed Central

Raeisi, Javad; Saifi, Mahnaz; Pourshafie, Mohammad Reza; Habibi, Mehri; Mohajerani, Hamid Reza; Akbari, Neda

2017-01-01

Introduction Vancomycin Resistant Enterococci (VRE) can be found all over the world. Thus, rapid detection of the isolates could be of high importance in the treatment or prevention of the associated disease. Aim To measure the turanose fermentation in Enterococcus faecalis clinical isolates for rapid differentiation of VRE and Vancomycin-Susceptible E. faecalis (VSE) isolates. Materials and Methods Forty E. faecalis samples were isolated from 200 clinical samples in Tehran Medical Center, Iran, from October 2012 to December 2012. These isolates were detected according to the standard microbial and biochemical tests. Detection of VRE isolates was originally performed by disk diffusion using 1 μg vancomycin disk, followed by Polymerase Chain Reaction (PCR) amplification of the vanA gene. Finally, the turanose consumption in 1%, 0.7% and 0.5% dilutions was detected by a phenotypic method. Results Among the 40 E. faecalis isolates, 20 vancomycin-susceptible and 20 vancomycin-resistant E. faecalis were isolated according to the disk diffusion and PCR of the vanA gene. There was a considerable difference between VRE and VSE isolates in 0.7% dilution of turanose. However, there was no significant difference between VRE and VSE in 1% and 0.5% dilutions of turanose. Conclusion Since detection of VRE isolates is of high importance, especially in nosocomial infections, phenotypic methods may be highly useful for this purpose. In conclusion, our data indicate that VRE isolated from clinical samples could be distinguished from VSE isolates by turanose fermentation at dilution 0.7%. PMID:28511382

Sponge-associated actinobacterial diversity: validation of the methods of actinobacterial DNA extraction and optimization of 16S rRNA gene amplification.

PubMed

Yang, Qi; Franco, Christopher M M; Zhang, Wei

2015-10-01

Experiments were designed to validate the two common DNA extraction protocols (CTAB-based method and DNeasy Blood & Tissue Kit) used to effectively recover actinobacterial DNA from sponge samples in order to study the sponge-associated actinobacterial diversity. This was done by artificially spiking sponge samples with actinobacteria (spores, mycelia and a combination of the two). Our results demonstrated that both DNA extraction methods were effective in obtaining DNA from the sponge samples as well as the sponge samples spiked with different amounts of actinobacteria. However, it was noted that in the presence of the sponge, the bacterial 16S rRNA gene could not be amplified unless the combined DNA template was diluted. To test the hypothesis that the extracted sponge DNA contained inhibitors, dilutions of the DNA extracts were tested for six sponge species representing five orders. The results suggested that the inhibitors were co-extracted with the sponge DNA, and a high dilution of this DNA was required for the successful PCR amplification for most of the samples. The optimized PCR conditions, including primer selection, PCR reaction system and program optimization, further improved the PCR performance. However, no single PCR condition was found to be suitable for the diverse sponge samples using various primer sets. These results highlight for the first time that the DNA extraction methods used are effective in obtaining actinobacterial DNA and that the presence of inhibitors in the sponge DNA requires high dilution coupled with fine tuning of the PCR conditions to achieve success in the study of sponge-associated actinobacterial diversity.

Development of an enzyme-linked immunosorbent assay for seven sulfonamide residues and investigation of matrix effects from different food samples.

PubMed

Zhang, Hongyan; Wang, Lei; Zhang, Yan; Fang, Guozhen; Zheng, Wenjie; Wang, Shuo

2007-03-21

Direct competitive enzyme-linked immunosorbent assays (ELISA) were developed to detect a broad range of sulfonamides in various matrices. Screening for this class of antibiotics in pig muscle, chicken muscle, fish, and egg extracts was accomplished by simple, rapid extraction methods carried out with only phosphate-buffered saline (PBS) buffer. Twenty milliliters of extract solution was added to 4 g of sample to extract the sulfonamide residues, and sample extracts diluted with assay buffer were directly analyzed by ELISA; matrix effects could be avoided with 1:5 dilution of pig muscle, chicken muscle, and egg extracts with PBS and 1:5 dilution of fish extract with 1% bovine serum albumin (BSA)-PBS. For liver sample, the extraction method was a little more complicated; 2 g of sample was added to 20 mL of ethanol, mixed, and then centrifuged. The solvent of 10 mL of the upper liquid was removed, and the residues were dissolved in 10 mL of PBS and then filtered; the filtrate was diluted two-fold with 0.5% BSA-PBS for ELISA. These common methods were able to detect seven sulfonamide residues such as sulfisozole, sulfathiazole, sufameter, sulfamethoxypyridazine, sulfapyridine, sulfamethizole, and sulfachlorpyridazine in pig muscle, liver, chicken muscle, egg, and fish. The assay's detection limits for these compounds were less than 100 microg kg-1. Various extraction methods were tested, and the average recovery (n=3) of 100 microg kg-1 for the matrices was found to range from 77.3 to 123.7%.

Comparison of susceptibility test methods to detect penicillin susceptibility in Streptococcus pneumoniae isolates.

PubMed

Mohd Nasir, Mohd Desa; Parasakthi, Navaratnam

2004-06-01

The increasing prevalence of penicillin-resistant Streptococuus pneumoniae urges for fast and accurate susceptibility testing methods. This study evaluated the comparability of three commonly used techniques; disk diffusion, E-test and agar dilution, to detect penicillin susceptibility in clinical isolates of S. pneumoniae. Fifty pneumococcal isolates, obtained from patients at the University of Malaya Medical Centre, were selected to include both penicillin-susceptible strains and those that had decreased susceptibility (resistant and intermediate) to penicillin. The minimum inhibitory concentration (MIC) values of penicillin to serve as the reference was determined by the agar dilution method in which, based on the MIC breakpoints recommended by the National Committee for Clinical Laboratory Standards (NCCLS), 27 strains had decreased susceptibility to penicillin with 17 strains resistant and 10 intermediate. Comparing to the agar dilution method, oxacillin disk diffusion test detected all strains with decreased penicillin susceptibility as such while E-test showed a close agreement of susceptibility (92%) of the isolates to penicillin. This confirmed that oxacillin is a good screening test for S. pneumoniae isolates with decreased susceptibility to penicillin while E-test is very reliable for rapid and accurate detection of penicillin susceptibility.

The measurement of lung volumes using body plethysmography and helium dilution methods in COPD patients: a correlation and diagnosis analysis.

PubMed

Tang, Yongjiang; Zhang, Mingke; Feng, Yulin; Liang, Binmiao

2016-11-23

Chronic obstructive pulmonary disease (COPD) is a chronic airway disease characterized by persistent airflow limitation. Moreover, lung hyperinflation evaluated by lung volumes is also the key pathophysiologic process during COPD progression. Nevertheless, there is still no preferred method to evaluate lung volumes. For this study, we recruited 170 patients with stable COPD to assess lung volumes stratified by airflow limitation severity. Lung volumes including residual volume (RV) and total lung capacity (TLC) were determined by both body plethysmography and helium dilution methods. The discrepancies between these two methods were recorded as ΔRV%pred, ΔTLC%pred, and ΔRV/TLC. We found that ΔRV%pred, ΔTLC%pred, and ΔRV/TLC increased significantly with the severity of COPD. The differences of lung capacity between these two methods were negatively correlated with FEV 1 %pred, and diffusing capacity for carbon monoxide (D L CO%pred). Moreover, the receiver operating characteristic (ROC) for ΔTLC%pred to distinguish severe COPD from non-severe COPD had an area under curve (AUC) of 0.886. The differences of lung volume parameters measured by body plethysmography and helium dilution methods were associated with airflow limitation and can effectively differentiate COPD severity, which may be a supportive method to assess the lung function of stable COPD patients.

Assessment of formulas for calculating critical concentration by the agar diffusion method.

PubMed Central

Drugeon, H B; Juvin, M E; Caillon, J; Courtieu, A L

1987-01-01

The critical concentration of antibiotic was calculated by using the agar diffusion method with disks containing different charges of antibiotic. It is currently possible to use different calculation formulas (based on Fick's law) devised by Cooper and Woodman (the best known) and by Vesterdal. The results obtained with the formulas were compared with the MIC results (obtained by the agar dilution method). A total of 91 strains and two cephalosporins (cefotaxime and ceftriaxone) were studied. The formula of Cooper and Woodman led to critical concentrations that were higher than the MIC, but concentrations obtained with the Vesterdal formula were closer to the MIC. The critical concentration was independent of method parameters (dilution, for example). PMID:3619419

Formation of nanocrystalline diamond in polymer like carbon films deposited by plasma CVD.

PubMed

Bhaduri, A; Chaudhuri, P

2009-09-01

Conventional plasma enhanced chemical vapour deposition (PECVD) method is generally not suitable for the growth of nanocrystalline diamond (NCD) films. However, our study shows that conditions favourable for powder formation help to grow large amount of nanocrystallites in conventional PECVD. With CH4 as the carbon source gas, dilution with Ar and moderate (50 W) rf power enhances formations of powders (nanoparticles) and C2 dimers within the plasma. On the other hand, with pure CH4 or with hydrogen diluted CH4, powder formation as also NCD growth is hindered. It is proposed that the nanoparticles formed in the plasma act as the "islands" while the C2 dimers are the "seeds" for the NCD growth. The structure of the films deposited on the grounded anode under different conditions of dilution has been studied. It is observed that with high Ar dilution the films contain NCD embedded in polymer like carbon (PLC) matrix.

Refractometry for quality control of anesthetic drug mixtures.

PubMed

Stabenow, Jennifer M; Maske, Mindy L; Vogler, George A

2006-07-01

Injectable anesthetic drugs used in rodents are often mixed and further diluted to increase the convenience and accuracy of dosing. We evaluated clinical refractometry as a simple and rapid method of quality control and mixing error detection of rodent anesthetic or analgesic mixtures. Dilutions of ketamine, xylazine, acepromazine, and buprenorphine were prepared with reagent-grade water to produce at least 4 concentration levels. The refraction of each concentration then was measured with a clinical refractometer and plotted against the percentage of stock concentration. The resulting graphs were linear and could be used to determine the concentration of single-drug dilutions or to predict the refraction of drug mixtures. We conclude that refractometry can be used to assess the concentration of dilutions of single drugs and can verify the mixing accuracy of drug combinations when the components of the mixture are known and fall within the detection range of the instrument.

Moderate Dilution of Copper Slag by Natural Gas

NASA Astrophysics Data System (ADS)

Zhang, Bao-jing; Zhang, Ting-an; Niu, Li-ping; Liu, Nan-song; Dou, Zhi-he; Li, Zhi-qiang

2018-01-01

To enable use of copper slag and extract the maximum value from the contained copper, an innovative method of reducing moderately diluted slag to smelt copper-containing antibacterial stainless steel is proposed. This work focused on moderate dilution of copper slag using natural gas. The thermodynamics of copper slag dilution and ternary phase diagrams of the slag system were calculated. The effects of blowing time, temperature, matte settling time, and calcium oxide addition were investigated. The optimum reaction conditions were identified to be blowing time of 20 min, reaction temperature of 1250°C, settling time of 60 min, CaO addition of 4% of mass of slag, natural gas flow rate of 80 mL/min, and outlet pressure of 0.1 MPa. Under these conditions, the Fe3O4 and copper contents of the residue were 7.36% and 0.50%, respectively.

Comparison of body composition assessment methods in pediatric intestinal failure.

PubMed

Mehta, Nilesh M; Raphael, Bram; Guteirrez, Ivan M; Quinn, Nicolle; Mitchell, Paul D; Litman, Heather J; Jaksic, Tom; Duggan, Christopher P

2014-07-01

The aim of the study was to examine the agreement of multifrequency bioelectric impedance analysis (BIA) and anthropometry with reference methods for body composition assessment in children with intestinal failure (IF). We conducted a prospective pilot study in children 14 years or younger with IF resulting from either short bowel syndrome or motility disorders. Bland-Altman analysis was used to examine the agreement between BIA and deuterium dilution in measuring total body water (TBW) and lean body mass (LBM), and between BIA and dual-energy x-ray absorptiometry (DXA) techniques in measuring LBM and fat mass (FM). FM and percent body fat (%BF) measurements by BIA and anthropometry were also compared in relation to those measured by deuterium dilution. Fifteen children with IF, median (interquartile range) age 7.2 (5.0, 10.0) years, and 10 (67%) boys, were studied. BIA and deuterium dilution were in good agreement with a mean bias (limits of agreement) of 0.9 (-3.2 to 5.0) for TBW (L) and 0.1 (-5.4 to 5.6) for LBM (kg) measurements. The mean bias (limits) for FM (kg) and %BF measurements were 0.4 (-3.8 to 4.6) kg and 1.7 (-16.9 to 20.3)%, respectively. The limits of agreement were within 1 standard deviation of the mean bias in 12 of 14 (86%) subjects for TBW and LBM, and in 11 of 14 (79%) for FM and %BF measurements. Mean bias (limits) for LBM (kg) and FM (kg) between BIA and DXA were 1.6 (-3.0 to 6.3) kg and -0.1 (-3.2 to 3.1) kg, respectively. Mean bias (limits) for FM (kg) and %BF between anthropometry and deuterium dilution were 0.2 (-4.2 to 4.6) and -0.2 (-19.5 to 19.1), respectively. The limits of agreement were within 1 standard deviation of the mean bias in 10 of 14 (71%) subjects. In children with IF, TBW and LBM measurements by multifrequency BIA method were in agreement with isotope dilution and DXA methods, with small mean bias and clinically acceptable limits of agreement. In comparison with deuterium dilution, BIA was comparable to anthropometry for FM and %BF assessments with small mean bias, but the limits of agreement were large. BIA is a reliable method for TBW and LBM assessments in population studies; however, its reliability in individual patients, especially for FM assessments, cannot be guaranteed.

Comparative study of SPORL and dilute-acid pretreatments of spruce for cellulosic ethanol production

Treesearch

L. Shuai; Q. Yang; Junyong Zhu; F.C. Lu; P.J. Weimer; J. Ralph; X.L. Pan

2010-01-01

The performance of two pretreatment methods, sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL) and dilute acid (DA), was compared in pretreating softwood (spruce) for fuel ethanol production at 180C for 30 min with a sulfuric acid loading of 5% on oven-dry wood and a 5:1 liquor to-wood ratio. SPORL was supplemented with 9% sodium...

In Vitro impairment of whole blood coagulation and platelet function by hypertonic saline hydroxyethyl starch

PubMed Central

2011-01-01

Background Hypertonic saline hydroxyethyl starch (HH) has been recommended for first line treatment of hemorrhagic shock. Its effects on coagulation are unclear. We studied in vitro effects of HH dilution on whole blood coagulation and platelet function. Furthermore 7.2% hypertonic saline, 6% hydroxyethylstarch (as ingredients of HH), and 0.9% saline solution (as control) were tested in comparable dilutions to estimate specific component effects of HH on coagulation. Methods The study was designed as experimental non-randomized comparative in vitro study. Following institutional review board approval and informed consent blood samples were taken from 10 healthy volunteers and diluted in vitro with either HH (HyperHaes®, Fresenius Kabi, Germany), hypertonic saline (HT, 7.2% NaCl), hydroxyethylstarch (HS, HAES6%, Fresenius Kabi, Germany) or NaCl 0.9% (ISO) in a proportion of 5%, 10%, 20% and 40%. Coagulation was studied in whole blood by rotation thrombelastometry (ROTEM) after thromboplastin activation without (ExTEM) and with inhibition of thrombocyte function by cytochalasin D (FibTEM), the latter was performed to determine fibrin polymerisation alone. Values are expressed as maximal clot firmness (MCF, [mm]) and clotting time (CT, [s]). Platelet aggregation was determined by impedance aggregrometry (Multiplate) after activation with thrombin receptor-activating peptide 6 (TRAP) and quantified by the area under the aggregation curve (AUC [aggregation units (AU)/min]). Scanning electron microscopy was performed to evaluate HyperHaes induced cell shape changes of thrombocytes. Statistics: 2-way ANOVA for repeated measurements, Bonferroni post hoc test, p < 0.01. Results Dilution impaired whole blood coagulation and thrombocyte aggregation in all dilutions in a dose dependent fashion. In contrast to dilution with ISO and HS, respectively, dilution with HH as well as HT almost abolished coagulation (MCFExTEM from 57.3 ± 4.9 mm (native) to 1.7 ± 2.2 mm (HH 40% dilution; p < 0.0001) and to 6.6 ± 3.4 mm (HT 40% dilution; p < 0.0001) and thrombocyte aggregation (AUC from 1067 ± 234 AU/mm (native) to 14.5 ± 12.5 AU/mm (HH 40% dilution; p < 0.0001) and to 20.4 ± 10.4 AU/min (HT 40% dilution; p < 0.0001) without differences between HH and HT (MCF: p = 0.452; AUC: p = 0.449). Conclusions HH impairs platelet function during in vitro dilution already at 5% dilution. Impairment of whole blood coagulation is significant after 10% dilution or more. This effect can be pinpointed to the platelet function impairing hypertonic saline component and to a lesser extend to fibrin polymerization inhibition by the colloid component or dilution effects. Accordingly, repeated administration and overdosage should be avoided. PMID:21310047

The integration of dilute acid hydrolysis of xylan and fast pyrolysis of glucan to obtain fermentable sugars.

PubMed

Jiang, Liqun; Wu, Nannan; Zheng, Anqing; Zhao, Zengli; He, Fang; Li, Haibin

2016-01-01

Fermentable sugars are important intermediates in the biological conversion of biomass. Hemicellulose and amorphous cellulose are easily hydrolyzed to fermentable sugars in dilute acid, whereas crystalline cellulose is more difficult to be hydrolyzed. Cellulose fast pyrolysis is an alternative method to liberate valuable fermentable sugars from biomass. The amount of levoglucosan generated from lignocellulose by fast pyrolysis is usually lower than the theoretical yield based on the cellulose fraction. Pretreatment is a promising route to improve the yield of levoglucosan from lignocellulose. The integration of dilute sulfuric acid hydrolysis and fast pyrolysis to obtain fermentable sugars was evaluated in this study. Dilute sulfuric acid hydrolysis could remove more than 95.1 and 93.4 % of xylan (the main component of hemicellulose) from sugarcane bagasse and corncob with high yield of xylose. On the other hand, dilute sulfuric acid hydrolysis was also an effective pretreatment to enhance levoglucosan yield from lignocellulose. Dilute acid hydrolysis could accumulate glucan (the component of cellulose) and remove most of the alkali and alkaline earth metals which were powerful catalysts during fast pyrolysis. Further increase in dilute acid concentration (from 0 to 2 %) in pretreatment could promote the yield of levoglucosan in fast pyrolysis. The acid pretreated sugarcane bagasse and corncob gave levoglucosan yields of 43.8 and 35.2 % which were obvious higher than those of raw sugarcane bagasse (12.0 %) and corncob (7.0 %). Obtaining fermentable sugars by combination dilute acid hydrolysis of xylan and fast pyrolysis of glucan could make full utilization of biomass, and get fermentable sugars economically from biomass for bio-refinery.

Multicenter Comparison of the Etest and EUCAST Methods for Antifungal Susceptibility Testing of Candida Isolates to Micafungin

PubMed Central

Bougnoux, M.-E.; Accoceberry, I.; Angoulvant, A.; Bailly, E.; Botterel, F.; Chevrier, S.; Chouaki, T.; Dalle, F.; Datry, A.; Dupuis, A.; Fekkar, A.; Gangneux, J. P.; Guitard, J.; Hennequin, C.; Le Govic, Y.; Le Pape, P.; Maubon, D.; Sautour, M.; Sendid, B.; Chandenier, J.

2016-01-01

In vitro susceptibility of 933 Candida isolates, from 16 French hospitals, to micafungin was determined using the Etest in each center. All isolates were then sent to a single center for determination of MICs by the EUCAST reference method. Overall essential agreement between the two tests was 98.5% at ±2 log2 dilutions and 90.2% at ±1 log2 dilutions. Categorical agreement was 98.2%. The Etest is a valuable alternative to EUCAST for the routine determination of micafungin MICs in medical mycology laboratories. PMID:27297480

Determination of atmospheric dilution for emergency preparedness: a joint EPA-DOE (Environmental Protection Agency-Department of Energy) technical workshop held on October 15-17, 1986

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

Rogers, S.E.

1988-10-01

This workshop was held in Research Triangle Park, NC in October of 1986. The objectives of the workshop were to review the current methods of determining atmospheric dilution for use in hazard identification, emergency-preparedness planning, and emergency response; to provide recommendations for choosing among these methods; and, finally, to define the role of the meteorologist in hazard identification, emergency planning, etc. Several invited papers were presented, and panel discussions were held to meet the objectives. The results are presented and discussed.

New SI-traceable reference gas mixtures for sulfur hexafluoride (SF6) at the pmol/mol level using static and dynamic preparation methods and comparison to existing scales

NASA Astrophysics Data System (ADS)

Wyss, Simon A.; Guillevic, Myriam; Vicar, Martin; Nieuwenkamp, Gerard; Vollmer, Martin K.; Pascale, Céline; Reimann, Stefan; Niederhauser, Bernhard; Emmenegger, Lukas

2017-04-01

We developed two SI-traceable methods, using both static and dynamic preparation steps, to produce reference gas mixtures for sulfur hexafluoride (SF6) in gas cylinders at pmol/mol level. This research activity is conducted under the framework of the European EMRP HIGHGAS project, in support of the high quality measurements of this important greenhouse gas in the earth's atmosphere. In the method used by the Czech Metrology Institute (CMI) a parent mixture of SF6 in synthetic air was produced in an aluminium cylinder at VSL as a first step. This mixture was produced gravimetrically according to ISO 6142 at an amount fraction of 1 μmol/mol. In the second step this primary standard was further diluted to near-ambient amount fraction, with the use of a three-step dilution system and directly pressurised into aluminium cylinders to a pressure of 10 bars. The second method used by the Federal Institute of Metrology (METAS) has already been applied to other fluorinated gases such as HFC-125 and HFC-1234yf. In this method a highly concentrated mixture is produced by spiking a purified synthetic air (matrix gas) with SF6 from a permeation device. The mass loss of SF6 in the permeation device is observed by a magnetic suspension balance. In a second step this mixture is diluted with matrix gas to the desired concentrations. All flows are controlled with mass flow controllers. The diluted gas was transferred into Silconert2000-coated stainless steel cylinders by cryo-filling. The final gas mixtures at near-ambient amount fraction were measured on a Medusa gas chromatography-mass spectrometry system (Medusa-GC/MS) against working standards calibrated on existing scales of the Scripps Institution of Oceanography (SIO) and compared to other scales [1]. The agreement of the assigned values by the CMI and METAS, with the measured values referenced on the SIO scale was excellent. This results show that with this methods we are able to produce accurate SI-traceable gas mixtures at near-ambient amount fraction for SF6, without extensive static dilutions. [1] Benjamin R. Miller, Ray F. Weiss, Peter K. Salameh, Toste Tanhua, Brian R. Greally, Jens Mühle, Peter G. Simmonds, Anal. Chem., 2008, 80, 1536.

Collaborative investigation of broth microdilution and semisolid agar dilution for in vitro susceptibility testing of Candida albicans.

PubMed Central

Shawar, R; Paetznick, V; Witte, Z; Ensign, L G; Anaissie, E; LaRocco, M

1992-01-01

A study was performed in two laboratories to evaluate the effect of growth medium and test methodology on inter- and intralaboratory variations in the MICs of amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITRA), and the triazole Sch 39304 (SCH) against 14 isolates of Candida albicans. Testing was performed by broth microdilution and semisolid agar dilution with the following media, buffered to pH 7.0 with morpholinepropanesulfonic acid (MOPS): buffered yeast nitrogen base (BYNB), Eagle's minimal essential medium (EMEM), RPMI 1640 medium (RPMI), and synthetic amino acid medium for fungi (SAAMF). Inocula were standardized spectrophotometrically, and endpoints were defined by the complete absence of growth for AMB and by no more than 25% of the growth in the drug-free control for all other agents. Comparative analyses of median MICs, as determined by each test method, were made for all drug-medium combinations. Both methods yielded similar (+/- 1 twofold dilution) median MICs for AMB in EMEM and RPMI, 5FC in all media, and FLU in EMEM, RPMI, and SAAMF. In contrast, substantial between-method variations in median MICs were seen for AMB in BYNB and SAAMF, FLU In BYNB, and ITRA and SCH in all media. Interlaboratory concordance of median MICs was good for AMB, 5FC, and FLU but poor for ITRA and SCH in all media. Endpoint determinations were analyzed by use of kappa statistical analyses for evaluating the strength of observer agreement. Moderate to almost perfect interlaboratory agreement occurred with AMB and 5FC in all media and with FLU in EMEM, RPMI, and SAAMF, irrespective of the test method. Slight to almost perfect interlaboratory agreement occurred with ITRA and SCH in EMEM, RPMI, and SAAMF when tested by semisolid agar dilution but not broth microdilution. Kappa values assessing intralaboratory agreement between methods were high for 5FC in all media, for AMB in BYNB, ENEM, and RPMI, and for FLU in EMEM, RPMI, and SAAMF. One laboratory, but not the other, reported substantial to almost perfect agreement between methods for ITRA, and SCH in EMEM, RPMI, and SAAMF. Both laboratories reported poor agreement between methods for the azoles in BYNB. Discrepancies noted in azole-BYNB combinations were largely due to the greater inhibitory effect of these agents in BYNB than in other media. These results indicate that the semisolid agar dilution and broth microdilution methods with EMEM or RPMI yield equivalent and reproducible MICs for AMB, 5FC, and FLU but not ITRA and SCH. PMID:1500502

Collaborative investigation of broth microdilution and semisolid agar dilution for in vitro susceptibility testing of Candida albicans.

PubMed

Shawar, R; Paetznick, V; Witte, Z; Ensign, L G; Anaissie, E; LaRocco, M

1992-08-01

A study was performed in two laboratories to evaluate the effect of growth medium and test methodology on inter- and intralaboratory variations in the MICs of amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITRA), and the triazole Sch 39304 (SCH) against 14 isolates of Candida albicans. Testing was performed by broth microdilution and semisolid agar dilution with the following media, buffered to pH 7.0 with morpholinepropanesulfonic acid (MOPS): buffered yeast nitrogen base (BYNB), Eagle's minimal essential medium (EMEM), RPMI 1640 medium (RPMI), and synthetic amino acid medium for fungi (SAAMF). Inocula were standardized spectrophotometrically, and endpoints were defined by the complete absence of growth for AMB and by no more than 25% of the growth in the drug-free control for all other agents. Comparative analyses of median MICs, as determined by each test method, were made for all drug-medium combinations. Both methods yielded similar (+/- 1 twofold dilution) median MICs for AMB in EMEM and RPMI, 5FC in all media, and FLU in EMEM, RPMI, and SAAMF. In contrast, substantial between-method variations in median MICs were seen for AMB in BYNB and SAAMF, FLU In BYNB, and ITRA and SCH in all media. Interlaboratory concordance of median MICs was good for AMB, 5FC, and FLU but poor for ITRA and SCH in all media. Endpoint determinations were analyzed by use of kappa statistical analyses for evaluating the strength of observer agreement. Moderate to almost perfect interlaboratory agreement occurred with AMB and 5FC in all media and with FLU in EMEM, RPMI, and SAAMF, irrespective of the test method. Slight to almost perfect interlaboratory agreement occurred with ITRA and SCH in EMEM, RPMI, and SAAMF when tested by semisolid agar dilution but not broth microdilution. Kappa values assessing intralaboratory agreement between methods were high for 5FC in all media, for AMB in BYNB, ENEM, and RPMI, and for FLU in EMEM, RPMI, and SAAMF. One laboratory, but not the other, reported substantial to almost perfect agreement between methods for ITRA, and SCH in EMEM, RPMI, and SAAMF. Both laboratories reported poor agreement between methods for the azoles in BYNB. Discrepancies noted in azole-BYNB combinations were largely due to the greater inhibitory effect of these agents in BYNB than in other media. These results indicate that the semisolid agar dilution and broth microdilution methods with EMEM or RPMI yield equivalent and reproducible MICs for AMB, 5FC, and FLU but not ITRA and SCH.

Thromboresistant surface coatings for the measurement of cardiac output through continuous low flow peripheral A-V shunts.

PubMed Central

Deeb, G M; Borovetz, H S; Griffith, B P; Hardesty, R L

1980-01-01

The dilution technique for determining cardiac output using indocyanine green dye is limited in patients weighing less than 20 kg because of the obligatory volume loss. Reproducible achieved using the green dye dilution method by the establishment of a low flow peripheral arteriovenous shunt. The shunt materials were treated with thromboresistant agents--TDMAC (7%) and albumin (1 g/dl)--to facilitate the use of this technique without heparin. For A-V shunt flow rates of 8-30 cc/min reproducible values of cardiac output were obtained for up to 38 hours which were in good agreement with determinations made using the conventional technique of dye dilution. PMID:6986120

Diluted thrombin time reliably measures low to intermediate plasma dabigatran concentrations.

PubMed

Božič-Mijovski, Mojca; Malmström, Rickard E; Malovrh, Petra; Antovic, Jovan P; Vene, Nina; Šinigoj, Petra; Mavri, Alenka

2016-07-01

Direct oral anticoagulant dabigatran was first introduced as a fixed-dose drug without routine coagulation monitoring, but current recommendations suggest that diluted thrombin time can be used to estimate plasma drug level. The aim of this study was to assess a diluted thrombin time assay based on the same thrombin reagent already used for traditional thrombin time measurements that reliably measure low to intermediate plasma dabigatran levels. We included 44 patients with atrial fibrillation who started treatment with dabigatran 150 mg (23 patients) or 110 mg (21 patients) twice a day. Blood samples were collected at baseline (no dabigatran) and 2-4 weeks after the beginning of dabigatran therapy at trough and at peak. Plasma dabigatran levels were measured with diluted thrombin time and compared to liquid chromatography with tandem mass spectrometry as the reference method. The performance of the diluted thrombin time was compared to Hemoclot® Thrombin Inhibitor and Ecarin Chromogenic Assay. In ex vivo plasma samples, diluted thrombin time highly correlated with the liquid chromatography with tandem mass spectrometry (Pearson's R = 0.9799). In the low to intermediate range (dabigatran concentration ≤ 100 µg/L) diluted thrombin time correlated significantly more closely to the liquid chromatography with tandem mass spectrometry (R = 0.964) than Hemoclot® Thrombin Inhibitor (R = 0.935, p = 0.05) or Ecarin Chromogenic Assay (R = 0.915, p < 0.01). It was also the only functional assay without any significant bias in the low to intermediate range. Both trough and peak diluted thrombin time values were similar to liquid chromatography with tandem mass spectrometry. We conclude that the diluted thrombin time assay presented in this study reliably detects dabigatran and that it is superior to the Hemoclot® Thrombin Inhibitor assay in the low to intermediate range. © The Author(s) 2015.

Estimation of the dilution field near a marine outfall by using effluent turbidity as an environmental tracer and comparison with dye tracer data.

PubMed

Pecly, José Otavio Goulart

2018-01-01

The alternative use of effluent turbidity to determine the dilution field of a domestic marine outfall located off the city of Rio de Janeiro was evaluated through field work comprising fluorescent dye tracer injection and tracking with simultaneous monitoring of sea water turbidity. A preliminary laboratory assessment was carried out with a sample of the outfall effluent whose turbidity was measured by the nephelometric method before and during a serial dilution process. During the field campaign, the dye tracer was monitored with field fluorometers and the turbidity was observed with an optical backscattering sensor interfaced to an OEM data acquisition system. About 4,000 samples were gathered, covering an area of 3 km × 3 km near the outfall diffusers. At the far field - where a drift towards the coastline was observed - the effluent plume was adequately labeled by the dye tracer. The turbidity plume was biased due to the high and variable background turbidity of sea water. After processing the turbidity dataset with a baseline detrending method, the plume presented high correlation with the dye tracer plume drawn on the near dilution field. However, dye tracer remains more robust than effluent turbidity.

Comparison of Dilution, Filtration, and Microwave Digestion Sample Pretreatments in Elemental Profiling of Wine by ICP-MS.

PubMed

Godshaw, Joshua; Hopfer, Helene; Nelson, Jenny; Ebeler, Susan E

2017-09-25

Wine elemental composition varies by cultivar, geographic origin, viticultural and enological practices, and is often used for authenticity validation. Elemental analysis of wine by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) is challenging due to the potential for non-spectral interferences and plasma instability arising from organic matrix components. Sample preparation mitigates these interferences, however, conflicting recommendations of best practices in ICP-MS analysis of wine have been reported. This study compared direct dilution, microwave-assisted acid digestion, and two filtration sample pretreatments, acidification prior to filtration and filtration followed by acidification, in elemental profiling of one white and three red table wines by ICP-MS. Of 43 monitored isotopes, 37 varied by sample preparation method, with significantly higher results of 17 isotopes in the microwave-digested samples. Both filtration treatments resulted in lower results for 11 isotopes compared to the other methods. Finally, isotope dilution determination of copper based on natural abundances and the 63 Cu: 65 Cu instrument response ratio agreed with external calibration and confirmed a significant sample preparation effect. Overall, microwave digestion did not compare favorably, and direct dilution was found to provide the best compromise between ease of use and result accuracy and precision, although all preparation strategies were able to differentiate the wines.

Dispensing Processes Impact Apparent Biological Activity as Determined by Computational and Statistical Analyses

PubMed Central

Ekins, Sean; Olechno, Joe; Williams, Antony J.

2013-01-01

Dispensing and dilution processes may profoundly influence estimates of biological activity of compounds. Published data show Ephrin type-B receptor 4 IC50 values obtained via tip-based serial dilution and dispensing versus acoustic dispensing with direct dilution differ by orders of magnitude with no correlation or ranking of datasets. We generated computational 3D pharmacophores based on data derived by both acoustic and tip-based transfer. The computed pharmacophores differ significantly depending upon dispensing and dilution methods. The acoustic dispensing-derived pharmacophore correctly identified active compounds in a subsequent test set where the tip-based method failed. Data from acoustic dispensing generates a pharmacophore containing two hydrophobic features, one hydrogen bond donor and one hydrogen bond acceptor. This is consistent with X-ray crystallography studies of ligand-protein interactions and automatically generated pharmacophores derived from this structural data. In contrast, the tip-based data suggest a pharmacophore with two hydrogen bond acceptors, one hydrogen bond donor and no hydrophobic features. This pharmacophore is inconsistent with the X-ray crystallographic studies and automatically generated pharmacophores. In short, traditional dispensing processes are another important source of error in high-throughput screening that impacts computational and statistical analyses. These findings have far-reaching implications in biological research. PMID:23658723

Hyperspectral imaging technique for detection of poultry fecal residues on food processing equipments

NASA Astrophysics Data System (ADS)

Cho, Byoung-Kwan; Kim, Moon S.; Chen, Yud-Ren

2005-11-01

Emerging concerns about safety and security in current mass production of food products necessitate rapid and reliable inspection for contaminant-free products. Diluted fecal residues on poultry processing plant equipment surface, not easily discernable from water by human eye, are contamination sources for poultry carcasses. Development of sensitive detection methods for fecal residues is essential to ensure safe production of poultry carcasses. Hyperspectral imaging techniques have shown good potential for detecting of the presence of fecal and other biological substances on food and processing equipment surfaces. In this study, use of high spatial resolution hyperspectral reflectance and fluorescence imaging (with UV-A excitation) is presented as a tool for selecting a few multispectral bands to detect diluted fecal and ingesta residues on materials used for manufacturing processing equipment. Reflectance and fluorescence imaging methods were compared for potential detection of a range of diluted fecal residues on the surfaces of processing plant equipment. Results showed that low concentrations of poultry feces and ingesta, diluted up to 1:100 by weight with double distilled water, could be detected using hyperspectral fluorescence images with an accuracy of 97.2%. Spectral bands determined in this study could be used for developing a real-time multispectral inspection device for detection of harmful organic residues on processing plant equipment.

Commutability of Cytomegalovirus WHO International Standard in Different Matrices

PubMed Central

Jones, Sara; Webb, Erika M.; Barry, Catherine P.; Choi, Won S.; Abravaya, Klara B.; Schneider, George J.

2016-01-01

Commutability of quantitative standards allows patient results to be compared across molecular diagnostic methods and laboratories. This is critical to establishing quantitative thresholds for use in clinical decision-making. A matrix effect associated with the 1st cytomegalovirus (CMV) WHO international standard (IS) was identified using the Abbott RealTime CMV assay. A commutability study was performed to compare the CMV WHO IS and patient specimens diluted in plasma and whole blood. Patient specimens showed similar CMV DNA quantitation values regardless of the diluent or extraction procedure used. The CMV WHO IS, on the other hand, exhibited a matrix effect. The CMV concentration reported for the WHO IS diluted in plasma was within the 95% prediction interval established with patient samples. In contrast, the reported DNA concentration of the CMV WHO IS diluted in whole blood was reduced approximately 0.4 log copies/ml, and values fell outside the 95% prediction interval. Calibrating the assay by using the CMV WHO IS diluted in whole blood would introduce a bias for CMV whole-blood quantitation; samples would be reported as having higher measured concentrations, by approximately 0.4 log IU/ml. Based on the commutability study with patient samples, the RealTime CMV assay was standardized based on the CMV WHO IS diluted in plasma. A revision of the instructions for use of the CMV WHO IS should be considered to alert users of the potential impact from the diluent matrix. The identification of a matrix effect with the CMV WHO IS underscores the importance of assessing commutability of the IS in order to achieve consistent results across methods. PMID:27030491

Study on the engine oil's wear based on the flash point

NASA Astrophysics Data System (ADS)

Niculescu, R.; Iorga-Simăn, V.; Trică, A.; Clenci, A.

2016-08-01

Increasing energy performance of internal combustion engines is largely influenced by frictional forces that arise between moving parts. Thus, in this respect, the nature and quality of the engine oil used is an important factor. Equally important is the effect of various engine injection strategies upon the oil quality. In other words, it's of utmost importance to maintain the quality of engine oil during engine's operation. Oil dilution is one of the most common causes that lead to its wear, creating lubrication problems. Moreover, at low temperatures operating conditions, the oil dilution with diesel fuel produces wax. When starting the engine, this may lead to lubrication deficiencies and even oil starvation with negative consequences on the engine mechanism parts wear (piston, rings and cylinders) but also crankcase bearings wear.Engine oil dilution with diesel fuel have several causes: wear of rings and/or injectors, late post-injection strategy for the sake of particulate filter regeneration, etc.This paper presents a study on the degree of deterioration of engine oils as a result of dilution with diesel fuel. The analysed oils used for this study were taken from various models of engines equipped with diesel particulate filter. The assessment is based on the determination of oil flash point and dilution degree using the apparatus Eraflash produced by Eralytics, Austria. Eraflash measurement is directly under the latest and safest standards ASTM D6450 & D7094), which are in excellent correlation with ASTM D93 Pensky - Martens ASTM D56 TAG methods; it uses the Continuous Closed Cup method for finding the Flash Point (CCCFP).

Method of quantitating dsDNA

DOEpatents

Stark, Peter C.; Kuske, Cheryl R.; Mullen, Kenneth I.

2002-01-01

A method for quantitating dsDNA in an aqueous sample solution containing an unknown amount of dsDNA. A first aqueous test solution containing a known amount of a fluorescent dye-dsDNA complex and at least one fluorescence-attenutating contaminant is prepared. The fluorescence intensity of the test solution is measured. The first test solution is diluted by a known amount to provide a second test solution having a known concentration of dsDNA. The fluorescence intensity of the second test solution is measured. Additional diluted test solutions are similarly prepared until a sufficiently dilute test solution having a known amount of dsDNA is prepared that has a fluorescence intensity that is not attenuated upon further dilution. The value of the maximum absorbance of this solution between 200-900 nanometers (nm), referred to herein as the threshold absorbance, is measured. A sample solution having an unknown amount of dsDNA and an absorbance identical to that of the sufficiently dilute test solution at the same chosen wavelength is prepared. Dye is then added to the sample solution to form the fluorescent dye-dsDNA-complex, after which the fluorescence intensity of the sample solution is measured and the quantity of dsDNA in the sample solution is determined. Once the threshold absorbance of a sample solution obtained from a particular environment has been determined, any similarly prepared sample solution taken from a similar environment and having the same value for the threshold absorbance can be quantified for dsDNA by adding a large excess of dye to the sample solution and measuring its fluorescence intensity.

Two techniques for eliminating luminol interference material and flow system configurations for luminol and firefly luciferase systems

NASA Technical Reports Server (NTRS)

Thomas, R. R.

1976-01-01

Two methods for eliminating luminol interference materials are described. One method eliminates interference from organic material by pre-reacting a sample with dilute hydrogen peroxide. The reaction rate resolution method for eliminating inorganic forms of interference is also described. The combination of the two methods makes the luminol system more specific for bacteria. Flow system designs for both the firefly luciferase and luminol bacteria detection systems are described. The firefly luciferase flow system incorporating nitric acid extraction and optimal dilutions has a functional sensitivity of 3 x 100,000 E. coli/ml. The luminol flow system incorporates the hydrogen peroxide pretreatment and the reaction rate resolution techniques for eliminating interference. The functional sensitivity of the luminol flow system is 1 x 10,000 E. coli/ml.

STUDIES ON ANALYTICAL METHODS FOR TRACE ELEMENTS IN METALS BY USING RADIOACTIVE ISOTOPE. III. DETERMINATION OF TANTALUM BY MEANS OF ISOTOPE DILUTION METHOD

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

Amano, H.

1959-10-01

The determination of tantalum by the isotope dilution method in the presence of niobium was investigated by the use of the radioisotope Ta/sup 185/. Tantalum was separated from niobium as tantalum-tannin precipitate under the optimum conditions of a pH of 1.9 to 2.5 and a tantalum/niobium ratio of up to 1/ 50. If niobium was present in amounts 100 times or more that of tantalum, reprecipitation was needed. The reciprocal of the specific activity of tanthlum pentoxide precipitate was in a linear relation to the change in the amount of tantalum added. The recommended method gave an accurate result inmore » the determination of tantalum in steal. (auth)« less

The efficacy of semi-quantitative urine protein-to-creatinine (P/C) ratio for the detection of significant proteinuria in urine specimens in health screening settings.

PubMed

Chang, Chih-Chun; Su, Ming-Jang; Ho, Jung-Li; Tsai, Yu-Hui; Tsai, Wei-Ting; Lee, Shu-Jene; Yen, Tzung-Hai; Chu, Fang-Yeh

2016-01-01

Urine protein detection could be underestimated using the conventional dipstick method because of variations in urine aliquots. This study aimed to assess the efficacy of the semi-quantitative urine protein-to-creatinine (P/C) ratio compared with other laboratory methods. Random urine samples were requested from patients undergoing chronic kidney disease screening. Significant proteinuria was determined by the quantitative P/C ratio of at least 150 mg protein/g creatinine. The semi-quantitative P/C ratio, dipstick protein and quantitative protein concentrations were compared and analyzed. In the 2932 urine aliquots, 156 (5.3 %) urine samples were considered as diluted and 60 (39.2 %) were found as significant proteinuria. The semi-quantitative P/C ratio testing had the best sensitivity (70.0 %) and specificity (95.9 %) as well as the lowest underestimation rate (0.37 %) when compared to other laboratory methods in the study. In the semi-quantitative P/C ratio test, 19 (12.2 %) had positive, 52 (33.3 %) had diluted, and 85 (54.5 %) had negative results. Of those with positive results, 7 (36.8 %) were positive detected by traditional dipstick urine protein test, and 9 (47.4 %) were positive detected by quantitative urine protein test. Additionally, of those with diluted results, 25 (48.1 %) had significant proteinuria, and all were assigned as no significant proteinuria by both tests. The semi-quantitative urine P/C ratio is clinically applicable based on its better sensitivity and screening ability for significant proteinuria than other laboratory methods, particularly in diluted urine samples. To establish an effective strategy for CKD prevention, urine protein screening with semi-quantitative P/C ratio could be considered.

[Formation of oxalate in oxaliplatin injection diluted with infusion solutions].

PubMed

Eto, Seiji; Yamamoto, Kie; Shimazu, Kounosuke; Sugiura, Toshimune; Baba, Kaori; Sato, Ayaka; Goromaru, Takeshi; Hagiwara, Yoshiaki; Hara, Keiko; Shinohara, Yoshitake; Takahashi, Kojiro

2014-01-01

Oxaliplatin use can cause acute peripheral neuropathy characterized by sensory paresthesias, which are markedly exacerbated by exposure to cold temperatures, and is a dose-limiting factor in the treatment of colorectal cancer.Oxalate is eliminated in a series of nonenzymatic conversions of oxaliplatin in infusion solutions or biological fluids.Elimination of oxalate from oxaliplatin has been suggested as one of the reasons for the development of acute neuropathy.In this study, we developed a high-performance liquid chromatography(HPLC)-based method to detect oxalate formation, and investigated the time dependent formation of oxalate in oxaliplatin diluted with infusion solutions.The results obtained showed that the amount of oxalate in the solution corresponded to 1.6% of oxaliplatin 8 h after oxaliplatin dilution with a 5% glucose solution. On the other hand, oxalate formation from oxaliplatin diluted with a saline solution was ten-fold higher than that from oxaliplatin diluted with the 5% glucose solution.Most patients who were intravenously injected with oxaliplatin experienced venous pain.As a preventive measure against venous pain, dexamethasone was added to the oxaliplatin injection.We measured the amount of oxalate formed in the dexamethasone-containing oxaliplatin injection diluted with a 5% glucose solution.The amount of oxalate formed when dexamethasone was added did not differ significantly from that formed when dexamethasone was not added.Thus, there are no clinical problems associated with the stability of oxaliplatin solutions.

A Study of the Vacancy-Impurity Interaction in Dilute Nickel Alloys by Core Electron Annihilation

NASA Astrophysics Data System (ADS)

Arbuzov, V. L.; Danilov, S. E.; Druzhkov, A. P.

1997-08-01

It is shown that the angular correlation of annihilation radiation can be used to identify vacancy-impurity complexes in dilute alloys. Annihilation of trapped positrons with core electrons bears information about the chemical environment of a vacancy defect. The method is especially effective for d-matrices doped with sp-impurities since annihilation parameters of positrons with d- and sp-shell electrons differ considerably. The potentialities of the method of core-electron annihilation of positrons are demonstrated taking electron-irradiated dilute Ni-P and Ni-Si alloys as an example. It is shown that the interaction between the vacancies, which migrate at the III stage of annealing, and P atoms in Ni-P causes a considerable change in the annihilation parameters of positrons with core electrons compared to pure Ni. In Ni-Si alloys the annihilation parameters of trapped positrons with core electrons do not differ from those in Ni. This fact is an evidence that Si atoms do not interact with vacancies in Ni.

Methods for detecting and correcting inaccurate results in inductively coupled plasma-atomic emission spectrometry

DOEpatents

Chan, George C. Y. [Bloomington, IN; Hieftje, Gary M [Bloomington, IN

2010-08-03

A method for detecting and correcting inaccurate results in inductively coupled plasma-atomic emission spectrometry (ICP-AES). ICP-AES analysis is performed across a plurality of selected locations in the plasma on an unknown sample, collecting the light intensity at one or more selected wavelengths of one or more sought-for analytes, creating a first dataset. The first dataset is then calibrated with a calibration dataset creating a calibrated first dataset curve. If the calibrated first dataset curve has a variability along the location within the plasma for a selected wavelength, errors are present. Plasma-related errors are then corrected by diluting the unknown sample and performing the same ICP-AES analysis on the diluted unknown sample creating a calibrated second dataset curve (accounting for the dilution) for the one or more sought-for analytes. The cross-over point of the calibrated dataset curves yields the corrected value (free from plasma related errors) for each sought-for analyte.

Inhibitory and bactericidal activities of levofloxacin, ofloxacin, erythromycin, and rifampin used singly and in combination against Legionella pneumophila.

PubMed Central

Baltch, A L; Smith, R P; Ritz, W

1995-01-01

The susceptibilities of 56 Legionella pneumophila isolates (43 clinical and 15 environmental isolates) to levofloxacin, ofloxacin, erythromycin, and rifampin were studied with buffered charcoal yeast extract (BCYE) agar (inoculum, 10(4) CFU per spot), and the susceptibilities of five isolates were studied with buffered yeast extract (BYE) broth (inoculum, 10(5) CFU/ml). The MICs inhibiting 90% of strains tested on BCYE agar were 0.125, 0.25, 1.0, and < or = 0.004 micrograms/ml for levofloxacin, ofloxacin, erythromycin, and rifampin, respectively. The MICs by the BYE broth dilution method were 1 to 3, 2, 1 to 2, and 1 tube lower than those by the agar dilution method for levofloxacin, ofloxacin, erythromycin, and rifampin, respectively. The MBCs were 1 to 2 tubes higher than the broth dilution MICs for levofloxacin, 1 to 3 tubes higher than the broth dilution MICs for ofloxacin, 1 to 3 tubes higher than the broth dilution MICs for erythromycin, and the same as the broth dilution MICs for rifampin. In kinetic time-kill curve studies, at drug concentrations of 1.0 and 2.0 times the MIC, the most active drugs were levofloxacin and rifampin. At 72 h, concentrations of levofloxacin and rifampin of 2.0 times the MIC demonstrated a bactericidal effect against L. pneumophila. In contrast, at concentrations of 1.0 and 2.0 times the MICs regrowth was observed with ofloxacin and only a gradual decrease in the numbers of CFU per milliliter was observed with erythromycin. Only a minor inhibitory effect was observed with 0.25 or 0.5 time the MICs of all drugs at 24 to 48 h, with regrowth occurring at 72 h. In contrast to erythromycin or ofloxacin plus rifampin at 0.25 time the MICs, only levofloxacin plus rifampin demonstrated synergy. Thus, levofloxacin demonstrated the best inhibitory and bactericidal effects against L. pneumophila when it was studied alone or in a combination with rifampin. PMID:7486896

Simultaneous determination of creatinine and creatine in human serum by double-spike isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC-MS).

PubMed

Fernández-Fernández, Mario; Rodríguez-González, Pablo; Añón Álvarez, M Elena; Rodríguez, Felix; Menéndez, Francisco V Álvarez; García Alonso, J Ignacio

2015-04-07

This work describes the first multiple spiking isotope dilution procedure for organic compounds using (13)C labeling. A double-spiking isotope dilution method capable of correcting and quantifying the creatine-creatinine interconversion occurring during the analytical determination of both compounds in human serum is presented. The determination of serum creatinine may be affected by the interconversion between creatine and creatinine during sample preparation or by inefficient chemical separation of those compounds by solid phase extraction (SPE). The methodology is based on the use differently labeled (13)C analogues ((13)C1-creatinine and (13)C2-creatine), the measurement of the isotopic distribution of creatine and creatinine by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the application of multiple linear regression. Five different lyophilized serum-based controls and two certified human serum reference materials (ERM-DA252a and ERM-DA253a) were analyzed to evaluate the accuracy and precision of the proposed double-spike LC-MS/MS method. The methodology was applied to study the creatine-creatinine interconversion during LC-MS/MS and gas chromatography-mass spectrometry (GC-MS) analyses and the separation efficiency of the SPE step required in the traditional gas chromatography-isotope dilution mass spectrometry (GC-IDMS) reference methods employed for the determination of serum creatinine. The analysis of real serum samples by GC-MS showed that creatine-creatinine separation by SPE can be a nonquantitative step that may induce creatinine overestimations up to 28% in samples containing high amounts of creatine. Also, a detectable conversion of creatine into creatinine was observed during sample preparation for LC-MS/MS. The developed double-spike LC-MS/MS improves the current state of the art for the determination of creatinine in human serum by isotope dilution mass spectrometry (IDMS), because corrections are made for all the possible errors derived from the sample preparation step.

A simple method for the comparison of commercially available ATP hygiene-monitoring systems.

PubMed

Colquhoun, K O; Timms, S; Fricker, C R

1998-04-01

The purpose of this study was to evaluate a methodology which could easily be used in any test laboratory in a uniform and consistent way for determining the sensitivity and reproducibility of results obtained with three ATP hygiene-monitoring systems. The test protocol discussed here allows such comparison to be made, thereby establishing a method of benchmarking both new systems and developments of existing systems. The sensitivity of the LUMINOMETER K, PocketSwab (Charm Sciences) was found to be between 0.4 and 4.0 nmol of ATP with poor reproducibility at the 40.0 nmol level (CV, 35%). The sensitivity of the IDEXX LIGHTING system and the Biotrace UNILITE Xcel were both between 0.04 and 0.4 nmol with coefficients of variation (CVs) of between 9% at 0.04 nmol and 10% at 0.4 nmol for the IDEXX system and 17% at 0.04 nmol and 21% at 0.4 nmol for the Biotrace system. The three systems were tested with a range of dilutions of different food residues: orange juice, raw milk, and ground beef slurry. All three test systems allowed detection of orange juice and raw milk at dilutions of 1:1,000, although the CV of results from the Charm system (54 and 74% respectively) was poor at this dilution for both residues. The sensitivity of the test systems was poorer for ground beef slurry than it was for orange juice and raw milk. Both the Biotrace and IDEXX systems were able to detect a 1:100 dilution of beef slurry (with CVs of 17 and 10% respectively), whilst at this dilution results from the Charm system had a CV of 55%. It was possible by using the method described in this paper to rank in order of sensitivity and reproducibility the three single-shot ATP hygiene-monitoring systems investigated, with the IDEXX LIGHTNING being the best, followed by the Biotrace UNILITE Xcel, and then the charm LUMINOMETER K, PocketSwab.

Echinocandin Susceptibility Testing of Candida Species: Comparison of EUCAST EDef 7.1, CLSI M27-A3, Etest, Disk Diffusion, and Agar Dilution Methods with RPMI and IsoSensitest Media▿

PubMed Central

Arendrup, Maiken Cavling; Garcia-Effron, Guillermo; Lass-Flörl, Cornelia; Lopez, Alicia Gomez; Rodriguez-Tudela, Juan-Luis; Cuenca-Estrella, Manuel; Perlin, David S.

2010-01-01

This study compared nine susceptibility testing methods and 12 endpoints for anidulafungin, caspofungin, and micafungin with the same collection of blinded FKS hot spot mutant (n = 29) and wild-type isolates (n = 94). The susceptibility tests included EUCAST Edef 7.1, agar dilution, Etest, and disk diffusion with RPMI-1640 plus 2% glucose (2G) and IsoSensitest-2G media and CLSI M27A-3. Microdilution plates were read after 24 and 48 h. The following test parameters were evaluated: fks hot spot mutants overlapping the wild-type distribution, distance between the two populations, number of very major errors (VMEs; fks mutants misclassified as susceptible), and major errors (MEs; wild-type isolates classified as resistant) using a wild-type-upper-limit value (WT-UL) (two twofold-dilutions higher than the MIC50) as the susceptibility breakpoint. The methods with the lowest number of errors (given as VMEs/MEs) across the three echinocandins were CLSI (12%/1%), agar dilution with RPMI-2G medium (14%/0%), and Etest with RPMI-2G medium (8%/3%). The fewest errors overall were observed for anidulafungin (4%/1% for EUCAST, 4%/3% for CLSI, and 3%/9% for Etest with RPMI-2G). For micafungin, VME rates of 10 to 71% were observed. For caspofungin, agar dilution with either medium was superior (VMEs/MEs of 0%/1%), while CLSI, EUCAST with IsoSensitest-2G medium, and Etest were less optimal (VMEs of 7%, 10%, and 10%, respectively). Applying the CLSI breakpoint (S ≤ 2 μg/ml) for CLSI results, 89.2% fks hot spot mutants were classified as anidulafungin susceptible, 60.7% as caspofungin susceptible, and 92.9% as micafungin susceptible. In conclusion, no test was perfect, but anidulafungin susceptibility testing using the WT-UL to define susceptibility reliably identified fks hot spot mutants. PMID:19884370

Designing and Interpreting Limiting Dilution Assays: General Principles and Applications to the Latent Reservoir for Human Immunodeficiency Virus-1.

PubMed

Rosenbloom, Daniel I S; Elliott, Oliver; Hill, Alison L; Henrich, Timothy J; Siliciano, Janet M; Siliciano, Robert F

2015-12-01

Limiting dilution assays are widely used in infectious disease research. These assays are crucial for current human immunodeficiency virus (HIV)-1 cure research in particular. In this study, we offer new tools to help investigators design and analyze dilution assays based on their specific research needs. Limiting dilution assays are commonly used to measure the extent of infection, and in the context of HIV they represent an essential tool for studying latency and potential curative strategies. Yet standard assay designs may not discern whether an intervention reduces an already miniscule latent infection. This review addresses challenges arising in this setting and in the general use of dilution assays. We illustrate the major statistical method for estimating frequency of infectious units from assay results, and we offer an online tool for computing this estimate. We recommend a procedure for customizing assay design to achieve desired sensitivity and precision goals, subject to experimental constraints. We consider experiments in which no viral outgrowth is observed and explain how using alternatives to viral outgrowth may make measurement of HIV latency more efficient. Finally, we discuss how biological complications, such as probabilistic growth of small infections, alter interpretations of experimental results.

Effects of iodinated contrast agent, xylocaine and gadolinium concentration on the signal emitted in magnetic resonance arthrography: a samples study*

PubMed Central

da Silva, Yvana Lopes Pinheiro; Costa, Rita Zanlorensi Visneck; Pinho, Kátia Elisa Prus; Ferreira, Ricardo Rabello; Schuindt, Sueliton Miyamoto

2015-01-01

Objective To investigate the effects of dilution of paramagnetic contrast agent with iodinated contrast and xylocaine on the signal intensity during magnetic resonance arthrography, and to improve the paramagnetic contrast agent concentration utilized in this imaging modality. Materials and Methods Samples specially prepared for the study with three different concentrations of paramagnetic contrast agent diluted in saline, iodinated contrast agent and xylocaine were imaged with fast spin echo T1-weighted sequences with fat saturation. The samples were placed into flasks and graphical analysis of the signal intensity was performed as a function of the paramagnetic contrast concentration. Results As compared with samples of equal concentrations diluted only with saline, the authors have observed an average signal intensity decrease of 20.67% for iodinated contrast agent, and of 28.34% for xylocaine. However, the increased gadolinium concentration in the samples caused decrease in signal intensity with all the dilutions. Conclusion Minimizing the use of iodinated contrast media and xylocaine and/or the use of a gadolinium concentration of 2.5 mmol/L diluted in saline will improve the sensitivity of magnetic resonance arthrography. PMID:25987746

One-step refolding and purification of recombinant human tumor necrosis factor-α (rhTNF-α) using ion-exchange chromatography.

PubMed

Wang, Yan; Ren, Wenxuan; Gao, Dong; Wang, Lili; Yang, Ying; Bai, Quan

2015-02-01

Protein refolding is a key step for the production of recombinant proteins, especially at large scales, and usually their yields are very low. Chromatographic-based protein refolding techniques have proven to be superior to conventional dilution refolding methods. High refolding yield can be achieved using these methods compared with dilution refolding of proteins. In this work, recombinant human tumor necrosis factor-α (rhTNF-α) from inclusion bodies expressed in Escherichia coli was renatured with simultaneous purification by ion exchange chromatography with a DEAE Sepharose FF column. Several chromatographic parameters influencing the refolding yield of the denatured/reduced rhTNF-α, such as the urea concentration, pH value and concentration ratio of glutathione/oxidized glutathione in the mobile phase, were investigated in detail. Under optimal conditions, rhTNF-α can be renatured and purified simultaneously within 30 min by one step. Specific bioactivity of 2.18 × 10(8) IU/mg, purity of 95.2% and mass recovery of 76.8% of refolded rhTNF-α were achieved. Compared with the usual dilution method, the ion exchange chromatography method developed here is simple and more effective for rhTNF-α refolding in terms of specific bioactivity and mass recovery. Copyright © 2014 John Wiley & Sons, Ltd.

Development of a micromanipulation method for single cell isolation of prokaryotes and its application in food safety.

PubMed

Hohnadel, Marisa; Maumy, Myriam; Chollet, Renaud

2018-01-01

For nearly a century, conventional microbiological methods have been standard practice for detecting and identifying pathogens in food. Nevertheless, the microbiological safety of food has improved and various rapid methods have been developed to overcome the limitations of conventional methods. Alternative methods are expected to detect low cell numbers, since the presence in food of even a single cell of a pathogenic organism may be infectious. With respect to low population levels, the performance of a detection method is assessed by producing serial dilutions of a pure bacterial suspension to inoculate representative food matrices with highly diluted bacterial cells (fewer than 10 CFU/ml). The accuracy of data obtained by multiple dilution techniques is not certain and does not exclude some colonies arising from clumps of cells. Micromanipulation techniques to capture and isolate single cells from environmental samples were introduced more than 40 years ago. The main limitation of the current micromanipulation technique is still the low recovery rate for the growth of a single cell in culture medium. In this study, we describe a new single cell isolation method and demonstrate that it can be used successfully to grow various types of microorganism from picked individual cells. Tests with Gram-positive and Gram-negative organisms, including cocci, rods, aerobes, anaerobes, yeasts and molds showed growth recovery rates from 60% to 100% after micromanipulation. We also highlight the use of our method to evaluate and challenge the detection limits of standard detection methods in food samples contaminated by a single cell of Salmonella enterica.

21 CFR 177.2440 - Polyethersulfone resins.

Code of Federal Regulations, 2014 CFR

2014-04-01

... determined by reduced viscosity in dimethyl formamide in accordance with ASTM method D2857-70 (Reapproved 1977), “Standard Test Method for Dilute Solution Viscosity of Polymers,” which is incorporated by...

Measurement of dispersion of nanoparticles in a dense suspension by high-sensitivity low-coherence dynamic light scattering

NASA Astrophysics Data System (ADS)

Ishii, Katsuhiro; Nakamura, Sohichiro; Sato, Yuki

2014-08-01

High-sensitivity low-coherence DLS apply to measurement of particle size distribution of pigments suspended in a ink. This method can be apply to extremely dense and turbid media without dilution. We show the temporal variation of particle size distribution of thixotropy and sedimentary pigments due to aggregation, agglomerate, and sedimentation. Moreover, we demonstrate the influence of dilution of ink to particle size distribution.

Development and Interlaboratory Study of a Liquid Chromatography Tandem Mass Spectrometric Methodfor the Determination of Multiple Mycotoxins inCereals Using Stable Isotope Dilution.

PubMed

Ye, Jin; Wu, Yu; Guo, Qilei; Lu, Meiling; Wang, Songshan; Xin, Yuanyuan; Xie, Gang; Zhang, Yan; Mariappan, Meena; Wang, Songxue

2018-05-01

An efficient, rapid, accurate, and cost-effective method based on stable isotope dilution and LC tandem MS was developed for the determination of multimycotoxins in cereals. The samples were extracted using acetonitrile-water-acetic acid (70 + 29 + 1, v/v/v), followed by dilution and centrifugation without any further cleanup. The mycotoxins were separated on a C18 column. Interference due to matrix effects was efficiently compensated for with [13C]-labeled stable isotope internal standards. The method demonstrated excellent linear relations, with regression coefficients above 0.999. Spiked recoveries at three different concentrations ranged from 80.9 to 115.9%, and RSDs were below 14% for all mycotoxins. The trueness of the method was also verified by participating in two proficiency tests, and satisfactory z-scores (|z| < 1.1) were obtained. In addition, an international interlaboratory study was organized to evaluate the methods. Eight laboratories characterized recovery, repeatability, and reproducibility studies in wheat, maize, and barley. The interlaboratory results were analyzed according to ISO 5725-2. Cochran and Grubbs tests were used to remove outliers. The mean recoveries of all 16 mycotoxins ranged from 87 to 111%. Repeatability, reproducibility, and Horwitz ratio values were 3.5-16.2, 5.4-33.6, and 0.16-1.65%, respectively. The results demonstrate that the method is reliable to determine multimycotoxins in cereals.

Most probable number methodology for quantifying dilute concentrations and fluxes of Escherichia coli O157:H7 in surface waters.

PubMed

Jenkins, M B; Endale, D M; Fisher, D S; Gay, P A

2009-02-01

To better understand the transport and enumeration of dilute densities of Escherichia coli O157:H7 in agricultural watersheds, we developed a culture-based, five tube-multiple dilution most probable number (MPN) method. The MPN method combined a filtration technique for large volumes of surface water with standard selective media, biochemical and immunological tests, and a TaqMan confirmation step. This method determined E. coli O157:H7 concentrations as low as 0.1 MPN per litre, with a 95% confidence level of 0.01-0.7 MPN per litre. Escherichia coli O157:H7 densities ranged from not detectable to 9 MPN per litre for pond inflow, from not detectable to 0.9 MPN per litre for pond outflow and from not detectable to 8.3 MPN per litre for within pond. The MPN methodology was extended to mass flux determinations. Fluxes of E. coli O157:H7 ranged from <27 to >10(4) MPN per hour. This culture-based method can detect small numbers of viable/culturable E. coli O157:H7 in surface waters of watersheds containing animal agriculture and wildlife. This MPN method will improve our understanding of the transport and fate of E. coli O157:H7 in agricultural watersheds, and can be the basis of collections of environmental E. coli O157:H7.

Multicenter Comparison of the Etest and EUCAST Methods for Antifungal Susceptibility Testing of Candida Isolates to Micafungin.

PubMed

Bougnoux, M-E; Dannaoui, E; Accoceberry, I; Angoulvant, A; Bailly, E; Botterel, F; Chevrier, S; Chouaki, T; Cornet, M; Dalle, F; Datry, A; Dupuis, A; Fekkar, A; Gangneux, J P; Guitard, J; Hennequin, C; Le Govic, Y; Le Pape, P; Maubon, D; Ranque, S; Sautour, M; Sendid, B; Chandenier, J

2016-08-01

In vitro susceptibility of 933 Candida isolates, from 16 French hospitals, to micafungin was determined using the Etest in each center. All isolates were then sent to a single center for determination of MICs by the EUCAST reference method. Overall essential agreement between the two tests was 98.5% at ±2 log2 dilutions and 90.2% at ±1 log2 dilutions. Categorical agreement was 98.2%. The Etest is a valuable alternative to EUCAST for the routine determination of micafungin MICs in medical mycology laboratories. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Faecal Parasitology: Concentration Methodology Needs to be Better Standardised

PubMed Central

Manser, Monika M.; Saez, Agatha Christie Santos; Chiodini, Peter L.

2016-01-01

Aim To determine whether variation in the preservative, pore size of the sieve, solvent, centrifugal force and centrifugation time used in the Ridley-Allen Concentration method for examining faecal specimens for parasite stages had any effect on their recovery in faecal specimens. Methods A questionnaire was sent to all participants in the UK NEQAS Faecal Parasitology Scheme. The recovery of parasite stages was compared using formalin diluted in water or formalin diluted in saline as the fixative, 3 different pore sizes of sieve, ether or ethyl acetate as a solvent, 7 different centrifugal forces and 6 different centrifugation times according to the methods described by participants completing the questionnaire. Results The number of parasite stages recovered was higher when formalin diluted in water was used as fixative, a smaller pore size of sieve was used, ethyl acetate along with Triton X 100 was used as a solvent and a centrifugal force of 3,000 rpm for 3 minutes were employed. Conclusions This study showed that differences in methodology at various stages of the concentration process affect the recovery of parasites from a faecal specimen and parasites present in small numbers could be missed if the recommended methodology is not followed. PMID:27073836

Antimicrobial Testing Methods & Procedures: MB-17-04

EPA Pesticide Factsheets

Information about ATMP - SOP Neutralization Confirmation Procedures for the AOAC Use-dilution method (UDM), the AOAC Germicidal Spray Products as Disinfectants Test (GSPT) and the Disinfectant Towelette Test (DTT) - MB-17-04

21 CFR 177.2440 - Polyethersulfone resins.

Code of Federal Regulations, 2013 CFR

2013-04-01

... molecular weight is determined by reduced viscosity in dimethyl formamide in accordance with ASTM method D2857-70 (Reapproved 1977), “Standard Test Method for Dilute Solution Viscosity of Polymers,” which is...

21 CFR 177.2440 - Polyethersulfone resins.

Code of Federal Regulations, 2012 CFR

2012-04-01

... molecular weight is determined by reduced viscosity in dimethyl formamide in accordance with ASTM method D2857-70 (Reapproved 1977), “Standard Test Method for Dilute Solution Viscosity of Polymers,” which is...

21 CFR 177.2440 - Polyethersulfone resins.

Code of Federal Regulations, 2011 CFR

2011-04-01

... molecular weight is determined by reduced viscosity in dimethyl formamide in accordance with ASTM method D2857-70 (Reapproved 1977), “Standard Test Method for Dilute Solution Viscosity of Polymers,” which is...

Rheology of dilute suspensions of red blood cells: experimental and theoretical approaches

NASA Astrophysics Data System (ADS)

Drochon, A.

2003-05-01

Shear viscosity measurements with dilute suspensions of red blood cells are interpreted using a microrheological model that relates the bulk measurements to the physical properties of the suspended cells. It is thus possible to quantify the average deformability of a RBC population in terms of a mean value of the membrane shear elastic modulus E_s. The values obtained for normal cells are in good agreement with those given in the literature. The method allows to discriminate between normal and altered (diamide or glutaraldehyde treated) cells or pathological cells (scleroderma). The predictions of the microrheological model, based on analytic calculations, are also compared with the numerical results of Ramanujan and Pozrikidis (JFM 361, 1998) for dilute suspensions of capsules in simple shear flow.

[Sampling methods for PM2.5 from stationary sources: a review].

PubMed

Jiang, Jing-Kun; Deng, Jian-Guo; Li, Zhen; Li, Xing-Hua; Duan, Lei; Hao, Ji-Ming

2014-05-01

The new China national ambient air quality standard has been published in 2012 and will be implemented in 2016. To meet the requirements in this new standard, monitoring and controlling PM2,,5 emission from stationary sources are very important. However, so far there is no national standard method on sampling PM2.5 from stationary sources. Different sampling methods for PM2.5 from stationary sources and relevant international standards were reviewed in this study. It includes the methods for PM2.5 sampling in flue gas and the methods for PM2.5 sampling after dilution. Both advantages and disadvantages of these sampling methods were discussed. For environmental management, the method for PM2.5 sampling in flue gas such as impactor and virtual impactor was suggested as a standard to determine filterable PM2.5. To evaluate environmental and health effects of PM2.5 from stationary sources, standard dilution method for sampling of total PM2.5 should be established.

Comparison of agar dilution and antibiotic gradient strip test with broth microdilution for susceptibility testing of swine Brachyspira species.

PubMed

Mirajkar, Nandita S; Gebhart, Connie J

2016-03-01

Production-limiting diseases in swine caused by Brachyspira are characterized by mucohemorrhagic diarrhea (B. hyodysenteriae and "B. hampsonii") or mild colitis (B. pilosicoli), while B. murdochii is often isolated from healthy pigs. Emergence of novel pathogenic Brachyspira species and strains with reduced susceptibility to commonly used antimicrobials has reinforced the need for standardized susceptibility testing. Two methods are currently used for Brachyspira susceptibility testing: agar dilution (AD) and broth microdilution (BMD). However, these tests have primarily been used for B. hyodysenteriae and rarely for B. pilosicoli. Information on the use of commercial susceptibility testing products such as antibiotic gradient strips is lacking. Our main objective was to validate and compare the susceptibility results, measured as the minimum inhibitory concentration (MIC), of 6 antimicrobials for 4 Brachyspira species (B. hyodysenteriae, "B. hampsonii", B. pilosicoli, and B. murdochii) by BMD and AD (tiamulin, valnemulin, lincomycin, tylosin, and carbadox) or antibiotic gradient strip (doxycycline) methods. In general, the results of a high percentage of all 4 Brachyspira species differed by ±1 log2 dilution or less by BMD and AD for tiamulin, valnemulin, lincomycin, and tylosin, and by BMD and antibiotic gradient strip for doxycycline. The carbadox MICs obtained by BMD were 1-5 doubling dilutions different than those obtained by AD. BMD for Brachyspira was quicker to perform with less ambiguous interpretation of results when compared with AD and antibiotic gradient strip methods, and the results confirm the utility of BMD in routine diagnostics. © 2016 The Author(s).

Irreproducible and uninterpretable Polymyxin B MICs for Enterobacter cloacae and Enterobacter aerogenes.

PubMed

Landman, David; Salamera, Julius; Quale, John

2013-12-01

Carbapenem-resistant Enterobacter species are emerging nosocomial pathogens. As with most multidrug-resistant Gram-negative pathogens, the polymyxins are often the only therapeutic option. In this study involving clinical isolates of E. cloacae and E. aerogenes, susceptibility testing methods with polymyxin B were analyzed. All isolates underwent testing by the broth microdilution (in duplicate) and agar dilution (in duplicate) methods, and select isolates were examined by the Etest method. Selected isolates were also examined for heteroresistance by population analysis profiling. Using a susceptibility breakpoint of ≤2 μg/ml, categorical agreement by all four dilution tests (two broth microdilution and two agar dilution) was achieved in only 76/114 (67%) of E. cloacae isolates (65 susceptible, 11 resistant). Thirty-eight (33%) had either conflicting or uninterpretable results (multiple skip wells, i.e., wells that exhibit no growth although growth does occur at higher concentrations). Of the 11 consistently resistant isolates, five had susceptible MICs as determined by Etest. Heteroresistant subpopulations were detected in eight of eight isolates tested, with greater percentages in isolates with uninterpretable MICs. For E. aerogenes, categorical agreement between the four dilution tests was obtained in 48/56 (86%), with conflicting and/or uninterpretable results in 8/56 (14%). For polymyxin susceptibility testing of Enterobacter species, close attention must be paid to the presence of multiple skip wells, leading to uninterpretable results. Susceptibility also should not be assumed based on the results of a single test. Until the clinical relevance of skip wells is defined, interpretation of polymyxin susceptibility tests for Enterobacter species should be undertaken with extreme caution.

Irreproducible and Uninterpretable Polymyxin B MICs for Enterobacter cloacae and Enterobacter aerogenes

PubMed Central

Landman, David; Salamera, Julius

2013-01-01

Carbapenem-resistant Enterobacter species are emerging nosocomial pathogens. As with most multidrug-resistant Gram-negative pathogens, the polymyxins are often the only therapeutic option. In this study involving clinical isolates of E. cloacae and E. aerogenes, susceptibility testing methods with polymyxin B were analyzed. All isolates underwent testing by the broth microdilution (in duplicate) and agar dilution (in duplicate) methods, and select isolates were examined by the Etest method. Selected isolates were also examined for heteroresistance by population analysis profiling. Using a susceptibility breakpoint of ≤2 μg/ml, categorical agreement by all four dilution tests (two broth microdilution and two agar dilution) was achieved in only 76/114 (67%) of E. cloacae isolates (65 susceptible, 11 resistant). Thirty-eight (33%) had either conflicting or uninterpretable results (multiple skip wells, i.e., wells that exhibit no growth although growth does occur at higher concentrations). Of the 11 consistently resistant isolates, five had susceptible MICs as determined by Etest. Heteroresistant subpopulations were detected in eight of eight isolates tested, with greater percentages in isolates with uninterpretable MICs. For E. aerogenes, categorical agreement between the four dilution tests was obtained in 48/56 (86%), with conflicting and/or uninterpretable results in 8/56 (14%). For polymyxin susceptibility testing of Enterobacter species, close attention must be paid to the presence of multiple skip wells, leading to uninterpretable results. Susceptibility also should not be assumed based on the results of a single test. Until the clinical relevance of skip wells is defined, interpretation of polymyxin susceptibility tests for Enterobacter species should be undertaken with extreme caution. PMID:24088860

In-vitro Antimicrobial Activities of Some Iranian Conifers

PubMed Central

Afsharzadeh, Maryam; Naderinasab, Mahboobe; Tayarani Najaran, Zahra; Barzin, Mohammad; Emami, Seyed Ahmad

2013-01-01

Male and female leaves and fruits of eleven different taxons of Iranian conifers (Cupressus sempervirens var. horizontalis, C. sempervirens var. sempervirens, C. sempervirens cv. Cereifeormis, Juniperus communis subsp. hemisphaerica, J. excelsa subsp. excelsa, J. excelsa subsp. polycarpos, J. foetidissima, J. oblonga, J. sabina, Platycladus orientalis and Taxus baccata) were collected from different localities of Iran, dried and extracted with methanol. The extracts were tested for their antimicrobial activity against Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Candida albicans. The extracts were screened qualitatively using four different methods, the disc diffusion, hole plate, cylinder agar diffusion and agar dilution methods, whereas the minimum inhibitory concentrations (MIC) of each extract were determined by the agar dilution method. The best result was obtained by means of hole plate method in qualitative determination of antimicrobial activities of extracts and the greatest activity was found against S. aureus in all tested methods. PMID:24250573

Quantification of integrated HIV DNA by repetitive-sampling Alu-HIV PCR on the basis of poisson statistics.

PubMed

De Spiegelaere, Ward; Malatinkova, Eva; Lynch, Lindsay; Van Nieuwerburgh, Filip; Messiaen, Peter; O'Doherty, Una; Vandekerckhove, Linos

2014-06-01

Quantification of integrated proviral HIV DNA by repetitive-sampling Alu-HIV PCR is a candidate virological tool to monitor the HIV reservoir in patients. However, the experimental procedures and data analysis of the assay are complex and hinder its widespread use. Here, we provide an improved and simplified data analysis method by adopting binomial and Poisson statistics. A modified analysis method on the basis of Poisson statistics was used to analyze the binomial data of positive and negative reactions from a 42-replicate Alu-HIV PCR by use of dilutions of an integration standard and on samples of 57 HIV-infected patients. Results were compared with the quantitative output of the previously described Alu-HIV PCR method. Poisson-based quantification of the Alu-HIV PCR was linearly correlated with the standard dilution series, indicating that absolute quantification with the Poisson method is a valid alternative for data analysis of repetitive-sampling Alu-HIV PCR data. Quantitative outputs of patient samples assessed by the Poisson method correlated with the previously described Alu-HIV PCR analysis, indicating that this method is a valid alternative for quantifying integrated HIV DNA. Poisson-based analysis of the Alu-HIV PCR data enables absolute quantification without the need of a standard dilution curve. Implementation of the CI estimation permits improved qualitative analysis of the data and provides a statistical basis for the required minimal number of technical replicates. © 2014 The American Association for Clinical Chemistry.

A dilute-and-shoot sample preparation strategy for new and used lubricating oils for Ca, P, S and Zn determination by total reflection X-ray fluorescence

NASA Astrophysics Data System (ADS)

Mota, Mariana F. B.; Gama, Ednilton M.; Rodrigues, Gabrielle de C.; Rodrigues, Guilherme D.; Nascentes, Clésia C.; Costa, Letícia M.

2018-01-01

In this work, a dilute-and-shoot method was developed for Ca, P, S and Zn determination in new and used lubricating oil samples by total reflection X-ray fluorescence (TXRF). The oil samples were diluted with organic solvents followed by addition of yttrium as internal standard and the TXRF measurements were performed after solvent evaporation. The method was optimized using an interlaboratorial reference material. The experimental parameters evaluated were sample volume (50 or 100 μL), measurement time (250 or 500 s) and volume deposited on the quartz glass sample carrier (5 or 10 μL). All of them were evaluated and optimized using xylene, kerosene and hexane. Analytical figures of merit (accuracy, precision, limit of detection and quantification) were used to evaluate the performance of the analytical method for all solvents. The recovery rates varied from 99 to 111% and the relative standard deviation remained between 1.7% and 10% (n = 8). For all elements, the results obtained by applying the new method were in agreement with the certified value. After the validation step, the method was applied for Ca, P, S and Zn quantification in eight new and four used lubricating oil samples, for all solvents. The concentration of the elements in the samples varied in the ranges of 1620-3711 mg L- 1 for Ca, 704-1277 mg L- 1 for P, 2027-9147 mg L- 1 for S, and 898-1593 mg L- 1 for Zn. The association of TXRF with a dilute-and-shoot sample preparation strategy was efficient for Ca, P, S and Zn determination in lubricating oils, presenting accurate results. Additionally, the time required for analysis is short, the reagent volumes are low minimizing waste generation, and the technique does not require calibration curves.

40 CFR 60.715 - Test methods and procedures.

Code of Federal Regulations, 2010 CFR

2010-07-01

... base film (i.e., the sample shall include any dilution solvent or other VOC added during the... for gas analysis. (f) Method 4 is used for stack gas moisture. (g) Methods 2, 2A, 2C, 2D, 3, and 4...

Ecological risk assessment of on-site soil washing with iron(III) chloride in cadmium-contaminated paddy field.

PubMed

Nagai, Takashi; Horio, Takeshi; Yokoyama, Atsushi; Kamiya, Takashi; Takano, Hiroyuki; Makino, Tomoyuki

2012-06-01

On-site soil washing with iron(III) chloride reduces Cd levels in soil, and thus the human health risks caused by Cd in food. However, it may threaten aquatic organisms when soil washing effluent is discharged to open aquatic systems. Therefore, we conducted trial-scale on-site soil washing and ecological risk assessment in Nagano and Niigata prefectures, Japan. The ecological effect of effluent water was investigated by two methods. The first was bioassay using standard aquatic test organisms. Twice-diluted effluent water from the Nagano site and the original effluent water from the Niigata site had no significant effects on green algae, water flea, caddisfly, and fish. The safe dilution rates were estimated as 20 times and 10 times for the Nagano and Niigata sites, respectively, considering an assessment factor of 10. The second method was probabilistic effect analysis using chemical analysis and the species sensitivity distribution concept. The mixture effects of CaCl(2), Al, Zn, and Mn were considered by applying a response additive model. The safe dilution rates, assessed for a potentially affected fraction of species of 5%, were 7.1 times and 23.6 times for the Nagano and Niigata sites, respectively. The actual dilution rates of effluent water by river water at the Nagano and Niigata sites were 2200-67,000 times and 1300-110,000 times, respectively. These are much larger than the safe dilution rates derived from the two approaches. Consequently, the ecological risk to aquatic organisms of soil washing is evaluated as being below the concern level. Copyright © 2012 Elsevier Inc. All rights reserved.

Dredging for dilution: A simulation based case study in a Tidal River.

PubMed

Bilgili, Ata; Proehl, Jeffrey A; Swift, M Robinson

2016-02-01

A 2-D hydrodynamic finite element model with a Lagrangian particle module is used to investigate the effects of dredging on the hydrodynamics and the horizontal dilution of pollutant particles originating from a wastewater treatment facility (WWTF) in tidal Oyster River in New Hampshire, USA. The model is driven by the semi-diurnal (M2) tidal component and includes the effect of flooding and drying of riverine mud flats. The particle tracking method consists of tidal advection plus a horizontal random walk model of sub-grid scale turbulent processes. Our approach is to perform continuous pollutant particle releases from the outfall, simulating three different scenarios: a base-case representing the present conditions and two different dredged channel/outfall location configurations. Hydrodynamics are investigated in an Eulerian framework and Lagrangian particle dilution improvement ratios are calculated for all cases. Results show that the simulated hydrodynamics are consistent with observed conditions. Eulerian and Lagrangian residuals predict an outward path suggesting flushing of pollutants on longer (>M2) time scales. Simulated dilution maps show that, in addition to dredging, the relocation of the WWTF outfall into the dredged main channel is required for increased dilution performance. The methodology presented here can be applied to similar managerial problems in all similar systems worldwide with relatively little effort, with the combination of Lagrangian and Eulerian methods working together towards a better solution. The statistical significance brought into methodology, by using a large number of particles (16000 in this case), is to be emphasized, especially with the growing number of networked parallel computer clusters worldwide. This paper improves on the study presented in Bilgili et al., 2006b, by adding an Eulerian analysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of pressure and fuel dilution on coflow laminar methane-air diffusion flames: A computational and experimental study

NASA Astrophysics Data System (ADS)

Cao, Su; Ma, Bin; Giassi, Davide; Bennett, Beth Anne V.; Long, Marshall B.; Smooke, Mitchell D.

2018-03-01

In this study, the influence of pressure and fuel dilution on the structure and geometry of coflow laminar methane-air diffusion flames is examined. A series of methane-fuelled, nitrogen-diluted flames has been investigated both computationally and experimentally, with pressure ranging from 1.0 to 2.7 atm and CH4 mole fraction ranging from 0.50 to 0.65. Computationally, the MC-Smooth vorticity-velocity formulation was employed to describe the reactive gaseous mixture, and soot evolution was modelled by sectional aerosol equations. The governing equations and boundary conditions were discretised on a two-dimensional computational domain by finite differences, and the resulting set of fully coupled, strongly nonlinear equations was solved simultaneously at all points using a damped, modified Newton's method. Experimentally, chemiluminescence measurements of CH* were taken to determine its relative concentration profile and the structure of the flame front. A thin-filament ratio pyrometry method using a colour digital camera was employed to determine the temperature profiles of the non-sooty, atmospheric pressure flames, while soot volume fraction was quantified, after evaluation of soot temperature, through an absolute light calibration using a thermocouple. For a broad spectrum of flames in atmospheric and elevated pressures, the computed and measured flame quantities were examined to characterise the influence of pressure and fuel dilution, and the major conclusions were as follows: (1) maximum temperature increases with increasing pressure or CH4 concentration; (2) lift-off height decreases significantly with increasing pressure, modified flame length is roughly independent of pressure, and flame radius decreases with pressure approximately as P-1/2; and (3) pressure and fuel stream dilution significantly affect the spatial distribution and the peak value of the soot volume fraction.

Rotor for processing liquids using movable capillary tubes

DOEpatents

Johnson, Wayne F.; Burtis, Carl A.; Walker, William A.

1989-01-01

A rotor assembly for processing liquids, especially whole blood samples, is disclosed. The assembly includes apparatus for separating non-liquid components of whole blood samples from liquid components, apparatus for diluting the separated liquid component with a diluent and apparatus for transferring the diluted sample to an external apparatus for analysis. The rotor assembly employs several movable capillary tubes to handle the sample and diluents. A method for using the rotor assembly to process liquids is also described.

Rotor for processing liquids using movable capillary tubes

DOEpatents

Johnson, Wayne F [Loudon, TN; Burtis, Carl A [Oak Ridge, TN; Walker, William A [Knoxville, TN

1989-05-30

A rotor assembly for processing liquids, especially whole blood samples, is disclosed. The assembly includes apparatus for separating non-liquid components of whole blood samples from liquid components, apparatus for diluting the separated liquid component with a diluent and apparatus for transferring the diluted sample to an external apparatus for analysis. The rotor assembly employs several movable capillary tubes to handle the sample and diluents. A method for using the rotor assembly to process liquids is also described.

Zn(1-x)MnxTe diluted magnetic semiconductor nanowires grown by molecular beam epitaxy.

PubMed

Zaleszczyk, Wojciech; Janik, Elzbieta; Presz, Adam; Dłuzewski, Piotr; Kret, Sławomir; Szuszkiewicz, Wojciech; Morhange, Jean-François; Dynowska, Elzbieta; Kirmse, Holm; Neumann, Wolfgang; Petroutchik, Aleksy; Baczewski, Lech T; Karczewski, Grzegorz; Wojtowicz, Tomasz

2008-11-01

It is shown that the growth of II-VI diluted magnetic semiconductor nanowires is possible by the catalytically enhanced molecular beam epitaxy (MBE). Zn(1-x)MnxTe NWs with manganese content up to x=0.60 were produced by this method. X-ray diffraction, Raman spectroscopy, and temperature dependent photoluminescence measurements confirm the incorporation of Mn(2+) ions in the cation substitutional sites of the ZnTe matrix of the NWs.

Development and Implementation of a Coagulation Factor Testing Method Utilizing Autoverification in a High-volume Clinical Reference Laboratory Environment.

PubMed

Riley, Paul W; Gallea, Benoit; Valcour, Andre

2017-01-01

Testing coagulation factor activities requires that multiple dilutions be assayed and analyzed to produce a single result. The slope of the line created by plotting measured factor concentration against sample dilution is evaluated to discern the presence of inhibitors giving rise to nonparallelism. Moreover, samples producing results on initial dilution falling outside the analytic measurement range of the assay must be tested at additional dilutions to produce reportable results. The complexity of this process has motivated a large clinical reference laboratory to develop advanced computer algorithms with automated reflex testing rules to complete coagulation factor analysis. A method was developed for autoverification of coagulation factor activity using expert rules developed with on an off the shelf commercially available data manager system integrated into an automated coagulation platform. Here, we present an approach allowing for the autoverification and reporting of factor activity results with greatly diminished technologist effort. To the best of our knowledge, this is the first report of its kind providing a detailed procedure for implementation of autoverification expert rules as applied to coagulation factor activity testing. Advantages of this system include ease of training for new operators, minimization of technologist time spent, reduction of staff fatigue, minimization of unnecessary reflex tests, optimization of turnaround time, and assurance of the consistency of the testing and reporting process.

Body Composition of Bangladeshi Children: Comparison and Development of Leg-to-Leg Bioelectrical Impedance Equation

PubMed Central

Khan, I.; Hawlader, Sophie Mohammad Delwer Hossain; Arifeen, Shams El; Moore, Sophie; Hills, Andrew P.; Wells, Jonathan C.; Persson, Lars-Åke; Kabir, Iqbal

2012-01-01

The aim of this study was to investigate the validity of the Tanita TBF 300A leg-to-leg bioimpedance analyzer for estimating fat-free mass (FFM) in Bangladeshi children aged 4-10 years and to develop novel prediction equations for use in this population, using deuterium dilution as the reference method. Two hundred Bangladeshi children were enrolled. The isotope dilution technique with deuterium oxide was used for estimation of total body water (TBW). FFM estimated by Tanita was compared with results of deuterium oxide dilution technique. Novel prediction equations were created for estimating FFM, using linear regression models, fitting child's height and impedance as predictors. There was a significant difference in FFM and percentage of body fat (BF%) between methods (p<0.01), Tanita underestimating TBW in boys (p=0.001) and underestimating BF% in girls (p<0.001). A basic linear regression model with height and impedance explained 83% of the variance in FFM estimated by deuterium oxide dilution technique. The best-fit equation to predict FFM from linear regression modelling was achieved by adding weight, sex, and age to the basic model, bringing the adjusted R2 to 89% (standard error=0.90, p<0.001). These data suggest Tanita analyzer may be a valid field-assessment technique in Bangladeshi children when using population-specific prediction equations, such as the ones developed here. PMID:23082630

Surface Dilution Kinetics Using Substrate Analog-Enantiomers as Diluents: Enzymatic Lipolysis by Bee-Venom Phospholipase A2

PubMed Central

Singh, Jasmeet; Ranganathan, Radha; Hajdu, Joseph

2010-01-01

A novel assay employing D-enantiomers of phospholipids as diluents for characterizing surface kinetics of lipid hydrolysis by phospholipases is introduced. The rationale of the method are: (i) D-enantiomers resist hydrolysis because of the stereoselectivity of the enzymes toward L-enantiomers and (ii) mixtures of L+D-lipids at various L:D ratios but constant L+D-lipid concentrations yield a surface dilution series of variable L-lipid concentration with constant medium properties. Kinetic characterization of bee-venom phospholipase A2 activity at bile salt + phospholipid aggregate-water interfaces was performed using the mixed L+D-lipid surface dilution assay and interface kinetic parameters were obtained. The assay applies to bio-membrane models as well. Activity was measured by pH-Stat methods. Aggregation numbers and interface hydration/microviscosity measured by time resolved fluorescence quenching and electron spin resonance respectively confirmed that interface properties were indeed invariant in a surface dilution series, supporting rationale (ii) and were used to calculate substrate concentrations. Activity data show excellent agreement with a kinetic model derived with D-enantiomers as diluents and also that D-phospholipids bind to the enzyme but resist hydrolysis; underscoring rationale (i). The assay is significant to enabling determination of interface specific kinetic parameters for the first time and thereby characterization of interface specificity of lipolytic enzymes. PMID:20727845

Estimation of the volatility distribution of organic aerosol combining thermodenuder and isothermal dilution measurements

NASA Astrophysics Data System (ADS)

Louvaris, Evangelos E.; Karnezi, Eleni; Kostenidou, Evangelia; Kaltsonoudis, Christos; Pandis, Spyros N.

2017-10-01

A method is developed following the work of Grieshop et al. (2009) for the determination of the organic aerosol (OA) volatility distribution combining thermodenuder (TD) and isothermal dilution measurements. The approach was tested in experiments that were conducted in a smog chamber using organic aerosol (OA) produced during meat charbroiling. A TD was operated at temperatures ranging from 25 to 250 °C with a 14 s centerline residence time coupled to a high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS) and a scanning mobility particle sizer (SMPS). In parallel, a dilution chamber filled with clean air was used to dilute isothermally the aerosol of the larger chamber by approximately a factor of 10. The OA mass fraction remaining was measured as a function of temperature in the TD and as a function of time in the isothermal dilution chamber. These two sets of measurements were used together to estimate the volatility distribution of the OA and its effective vaporization enthalpy and accommodation coefficient. In the isothermal dilution experiments approximately 20 % of the OA evaporated within 15 min. Almost all the OA evaporated in the TD at approximately 200 °C. The resulting volatility distributions suggested that around 60-75 % of the cooking OA (COA) at concentrations around 500 µg m-3 consisted of low-volatility organic compounds (LVOCs), 20-30 % of semivolatile organic compounds (SVOCs), and around 10 % of intermediate-volatility organic compounds (IVOCs). The estimated effective vaporization enthalpy of COA was 100 ± 20 kJ mol-1 and the effective accommodation coefficient was 0.06-0.07. Addition of the dilution measurements to the TD data results in a lower uncertainty of the estimated vaporization enthalpy as well as the SVOC content of the OA.

21 CFR 177.1560 - Polyarylsulfone resins.

Code of Federal Regulations, 2014 CFR

2014-04-01

... reduced viscosity of 0.40 deciliter per gram in 1-methyl-2-pyrrolidinone in accordance with ASTM method D2857-70 (Reapproved 1977), “Standard Test Method for Dilute Solution Viscosity of Polymers,” which is...

Evaluating an alternative method for rapid urinary creatinine determination

EPA Science Inventory

Creatinine (CR) is an endogenously-produced chemical routinely assayed in urine specimens to assess kidney function, sample dilution. The industry-standard method for CR determination, known as the kinetic Jaffe (KJ) method, relies on an exponential rate of a colorimetric change,...

Determination of dilution factors for discharge of aluminum-containing wastes by public water-supply treatment facilities into lakes and reservoirs in Massachusetts

USGS Publications Warehouse

Colman, John A.; Massey, Andrew J.; Brandt, Sara L.

2011-09-16

Dilution of aluminum discharged to reservoirs in filter-backwash effluents at water-treatment facilities in Massachusetts was investigated by a field study and computer simulation. Determination of dilution is needed so that permits for discharge ensure compliance with water-quality standards for aquatic life. The U.S. Environmental Protection Agency chronic standard for aluminum, 87 micrograms per liter (μg/L), rather than the acute standard, 750 μg/L, was used in this investigation because the time scales of chronic exposure (days) more nearly match rates of change in reservoir concentrations than do the time scales of acute exposure (hours).Whereas dilution factors are routinely computed for effluents discharged to streams solely on the basis of flow of the effluent and flow of the receiving stream, dilution determination for effluents discharged to reservoirs is more complex because (1), compared to streams, additional water is available for dilution in reservoirs during low flows as a result of reservoir flushing and storage during higher flows, and (2) aluminum removal in reservoirs occurs by aluminum sedimentation during the residence time of water in the reservoir. Possible resuspension of settled aluminum was not considered in this investigation. An additional concern for setting discharge standards is the substantial concentration of aluminum that can be naturally present in ambient surface waters, usually in association with dissolved organic carbon (DOC), which can bind aluminum and keep it in solution.A method for dilution determination was developed using a mass-balance equation for aluminum and considering sources of aluminum from groundwater, surface water, and filter-backwash effluents and losses caused by sedimentation, water withdrawal, and spill discharge from the reservoir. The method was applied to 13 reservoirs. Data on aluminum and DOC concentrations in reservoirs and influent water were collected during the fall of 2009. Complete reservoir volume was determined to be available for mixing on the basis of vertical and horizontal aluminum-concentration profiling. Losses caused by settling of aluminum were assumed to be proportional to aluminum concentration and reservoir area. The constant of proportionality, as a function of DOC concentration, was established by simulations in each of five reservoirs that differed in DOC concentration.In addition to computing dilution factors, the project determined dilution factors that would be protective with the same statistical basis (frequency of exceedance of the chronic standard) as dilutions computed for streams at the 7-day-average 10-year-recurrence annual low flow (the 7Q10). Low-flow dilutions are used for permitting so that receiving waters are protected even at the worst-case flow levels. The low-flow dilution factors that give the same statistical protection are the lowest annual 7-day-average dilution factors with a recurrence of 10 years, termed 7DF10s. Determination of 7DF10 values for reservoirs required that long periods of record be simulated so that dilution statistics could be determined. Dilution statistics were simulated for 13 reservoirs from 1960 to 2004 using U.S. Geological Survey Firm-Yield Estimator software to model reservoir inputs and outputs and present-day values of filter-effluent discharge and aluminum concentration.Computed settling velocities ranged from 0 centimeters per day (cm/d) at DOC concentrations of 15.5 milligrams per liter (mg/L) to 21.5 cm/d at DOC concentrations of 2.7 mg/L. The 7DF10 values were a function of aluminum effluent discharged. At current (2009) effluent discharge rates, the 7DF10 values varied from 1.8 to 115 among the 13 reservoirs. In most cases, the present-day (2009) discharge resulted in receiving water concentrations that did not exceed the standard at the 7DF10. Exceptions were one reservoir with a very small area and three reservoirs with high concentrations of DOC. Maximum permissible discharges were determined for water-treatment plants by adjusting discharges upward in simulations until the 7DF10 resulted in reservoir concentrations that just met the standard. In terms of aluminum flux, these discharges ranged from 0 to 28 kilograms of aluminum per day.

Distribution of the hallucinogens N,N-dimethyltryptamine and 5-methoxy-N,N-dimethyltryptamine in rat brain following intraperitoneal injection: application of a new solid-phase extraction LC-APcI-MS-MS-isotope dilution method.

PubMed

Barker, S A; Littlefield-Chabaud, M A; David, C

2001-02-10

A method for the solid-phase extraction (SPE) and liquid chromatographic-atmospheric pressure chemical ionization-mass spectrometric-mass spectrometric-isotope dilution (LC-APcI-MS-MS-ID) analysis of the indole hallucinogens N,N-dimethyltryptamine (DMT) and 5-methoxy DMT (or O-methyl bufotenin, OMB) from rat brain tissue is reported. Rats were administered DMT or OMB by the intraperitoneal route at a dose of 5 mg/kg and sacrificed 15 min post treatment. Brains were dissected into discrete areas and analyzed by the methods described as a demonstration of the procedure's applicability. The synthesis and use of two new deuterated internal standards for these purposes are also reported.

Principles, performance, and applications of spectral reconstitution (SR) in quantitative analysis of oils by Fourier transform infrared spectroscopy (FT-IR).

PubMed

García-González, Diego L; Sedman, Jacqueline; van de Voort, Frederik R

2013-04-01

Spectral reconstitution (SR) is a dilution technique developed to facilitate the rapid, automated, and quantitative analysis of viscous oil samples by Fourier transform infrared spectroscopy (FT-IR). This technique involves determining the dilution factor through measurement of an absorption band of a suitable spectral marker added to the diluent, and then spectrally removing the diluent from the sample and multiplying the resulting spectrum to compensate for the effect of dilution on the band intensities. The facsimile spectrum of the neat oil thus obtained can then be qualitatively or quantitatively analyzed for the parameter(s) of interest. The quantitative performance of the SR technique was examined with two transition-metal carbonyl complexes as spectral markers, chromium hexacarbonyl and methylcyclopentadienyl manganese tricarbonyl. The estimation of the volume fraction (VF) of the diluent in a model system, consisting of canola oil diluted to various extents with odorless mineral spirits, served as the basis for assessment of these markers. The relationship between the VF estimates and the true volume fraction (VF(t)) was found to be strongly dependent on the dilution ratio and also depended, to a lesser extent, on the spectral resolution. These dependences are attributable to the effect of changes in matrix polarity on the bandwidth of the ν(CO) marker bands. Excellent VF(t) estimates were obtained by making a polarity correction devised with a variance-spectrum-delineated correction equation. In the absence of such a correction, SR was shown to introduce only a minor and constant bias, provided that polarity differences among all the diluted samples analyzed were minimal. This bias can be built into the calibration of a quantitative FT-IR analytical method by subjecting appropriate calibration standards to the same SR procedure as the samples to be analyzed. The primary purpose of the SR technique is to simplify preparation of diluted samples such that only approximate proportions need to be adhered to, rather than using exact weights or volumes, the marker accounting for minor variations. Additional applications discussed include the use of the SR technique in extraction-based, quantitative, automated FT-IR methods for the determination of moisture, acid number, and base number in lubricating oils, as well as of moisture content in edible oils.

Nuclei-mode particulate emissions and their response to fuel sulfur content and primary dilution during transient operations of old and modern diesel engines.

PubMed

Liu, Z Gerald; Vasys, Victoria N; Kittelson, David B

2007-09-15

The effects of fuel sulfur content and primary dilution on PM number emissions were investigated during transient operations of an old and a modern diesel engine. Emissions were also studied during steady-state operations in order to confirm consistency with previous findings. Testing methods were concurrent with those implemented by the EPA to regulate PM mass emissions, including the use of the Federal Transient Testing Procedure-Heavy Duty cycle to simulate transient conditions and the use of a Critical Flow Venturi-Constant Volume System to provide primary dilution. Steady-state results were found to be consistent with previous studies in that nuclei-mode particulate emissions were largely reduced when lower-sulfur content fuel was used in the newer engine, while the nuclei-mode PM emissions from the older engine were much less affected by fuel sulfur content. The transient results, however, show that the total number of nuclei-mode PM emissions from both engines increases with fuel sulfur content, although this effect is only seen under the higher primary dilution ratios with the older engine. Transient results further show that higher primary dilution ratios increase total nuclei-mode PM number emissions in both engines.

Indirect enzyme-linked immunosorbent assay method based on Streptococcus agalactiae rSip-Pgk-FbsA fusion protein for detection of bovine mastitis.

PubMed

Bu, Ri-E; Wang, Jin-Liang; Wu, Jin-Hua; Xilin, Gao-Wa; Chen, Jin-Long; Wang, Hua

2017-03-01

The aim of this study was to establish a rapid and accurate method for the detection of the Streptococcus agalactiae antibody (SA-Ab) to determine the presence of the bovine mastitis (BM)-causative pathogen. The multi-subunit fusion protein rSip-Pgk-FbsA was prokaryotically expressed and purified. The triple activities of the membrane surface-associated proteins Sip, phosphoglycerate kinase (Pgk), and fibronectin (FbsA) were used as the diagnostic antigens to establish an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of SA-Ab in BM. The optimal antigen coating concentration was 2 μg/mL, the optimal serum dilution was 1:160, and the optimal dilution of the enzyme-labeled secondary antibody was 1:6000. The sensitivity, specificity, and repeatability tests showed that the method established in this study had no cross-reaction with antibodies to Streptococcus pyogenes, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis in the sera. The results of the sensitivity test showed that a positive result could be obtained even if the serum dilution reached 1:12,800, indicating the high sensitivity and good repeatability of the method. The positive coincidence rate of this method was 98.6%, which is higher than that of previous tests established with the Sip or Pgk mono-antigen fusion protein, respectively, demonstrating the relatively higher sensitivity of this newly established method. The detection rate for 389 clinical samples was 46.53%. The indirect ELISA method established in this study could provide a more accurate and reliable serological method for the rapid detection of S. agalactiae in cases of BM.

Ab initio description of the diluted magnetic semiconductor Ga1-xMnxAs: Ferromagnetism, electronic structure, and optical response

NASA Astrophysics Data System (ADS)

Craco, L.; Laad, M. S.; Müller-Hartmann, E.

2003-12-01

Motivated by a study of various experiments describing the electronic and magnetic properties of the diluted magnetic semiconductor Ga1-xMnxAs, we investigate its physical response in detail using a combination of first-principles band structure with methods based on dynamical mean field theory to incorporate strong, dynamical correlations, and intrinsic as well as extrinsic disorder in one single theoretical picture. We show how ferromagnetism is driven by double exchange (DE), in agreement with very recent observations, along with a good quantitative description of the details of the electronic structure, as probed by scanning tunneling microscopy and optical conductivity. Our results show how ferromagnetism can be driven by DE even in diluted magnetic semiconductors with small carrier concentration.

Evaluation of Mediterranean Agricultural Residues as a Potential Feedstock for the Production of Biogas via Anaerobic Fermentation.

PubMed

Nitsos, Christos; Matsakas, Leonidas; Triantafyllidis, Kostas; Rova, Ulrika; Christakopoulos, Paul

2015-01-01

Hydrothermal, dilute acid, and steam explosion pretreatment methods, were evaluated for their efficiency to improve the methane production yield of three Mediterranean agricultural lignocellulosic residues such as olive tree pruning, grapevine pruning, and almond shells. Hydrothermal and dilute acid pretreatments provided low to moderate increase in the digestibility of the biomass samples, whereas steam explosion enabled the highest methane yields to be achieved for almond shells at 232.2 ± 13.0 mL CH4/gVS and olive pruning at 315.4 ± 0.0 mL CH4/gVS. Introduction of an enzymatic prehydrolysis step moderately improved methane yields for hydrothermal and dilute acid pretreated samples but not for the steam exploded ones.

Evaluation of Mediterranean Agricultural Residues as a Potential Feedstock for the Production of Biogas via Anaerobic Fermentation

PubMed Central

Nitsos, Christos; Triantafyllidis, Kostas

2015-01-01

Hydrothermal, dilute acid, and steam explosion pretreatment methods, were evaluated for their efficiency to improve the methane production yield of three Mediterranean agricultural lignocellulosic residues such as olive tree pruning, grapevine pruning, and almond shells. Hydrothermal and dilute acid pretreatments provided low to moderate increase in the digestibility of the biomass samples, whereas steam explosion enabled the highest methane yields to be achieved for almond shells at 232.2 ± 13.0 mL CH4/gVS and olive pruning at 315.4 ± 0.0 mL CH4/gVS. Introduction of an enzymatic prehydrolysis step moderately improved methane yields for hydrothermal and dilute acid pretreated samples but not for the steam exploded ones. PMID:26609521

Biomimetic materials for protein storage and transport

DOEpatents

Firestone, Millicent A [Elmhurst, IL; Laible, Philip D [Villa Park, IL

2012-05-01

The invention provides a method for the insertion of protein in storage vehicles and the recovery of the proteins from the vehicles, the method comprising supplying isolated protein; mixing the isolated protein with a fluid so as to form a mixture, the fluid comprising saturated phospholipids, lipopolymers, and a surfactant; cycling the mixture between a first temperature and a second temperature; maintaining the mixture as a solid for an indefinite period of time; diluting the mixture in detergent buffer so as to disrupt the composition of the mixture, and diluting to disrupt the fluid in its low viscosity state for removal of the guest molecules by, for example, dialysis, filtering or chromatography dialyzing/filtering the emulsified solid.

[Epidemiologic diagnostic of nosocomial suppurative-septic infections of Pseudomonas etiology based on intraspecies typing of causative agent].

PubMed

Fel'dblium, I V; Zakharova, Iu A; Nikolaeva, A M; Fedotova, O S

2013-01-01

Scientific justification of optimization of epidemiologic diagnostic of suppurative-septic infection (SSI) caused by Pseudomonas aeruginosa based on comparability of antibiotic sensitivity and beta-lactamase production. Intraspecies typing of 37 P. aeruginosa strains isolated during microbiological monitoring of 106 patients and 131 objects of clinical environment of surgical and obstetrician hospitals by using a complex ofphenotypic and molecular-biological methods including determination of sensitivity to antibiotics by serial dilutions method and PCR-diagnostics with determination of TEM, SHV, CTX, OXA, MBL, VIM genes was performed. P. aeruginosa strains combined into groups by isolation location during studies turned out to be heterogeneous by sensitivity to antibiotics and beta-lactamase production that allowed to form subgroups of strains by focality attribute. Isolates recovered from different SSI foci had significant differences in minimal inhibitory concentration (MIC) reaching 1024 times. MIC parameter within subgroups did not exceed 8 - 16 consequent dilutions. Use of a complex of phenotypic and molecular-biologic methods of causative agent typing including determination of sensitivity to antibiotics by serial dilutions method and evaluation of beta-lactamase production allowed to establish a mechanism of development of SSI epidemic process caused by P. aeruginosa, detect origins and reservoirs of infection in hospital, modes and factors of transmission and reach maximum justification of epidemiologic control and prophylaxis measures of localization of foci of nosocomial infections of pseudomonas etiology.

Validation of highly sensitive simultaneous targeted and untargeted analysis of keto-steroids by Girard P derivatization and stable isotope dilution-liquid chromatography-high resolution mass spectrometry.

PubMed

Frey, Alexander J; Wang, Qingqing; Busch, Christine; Feldman, Daniel; Bottalico, Lisa; Mesaros, Clementina A; Blair, Ian A; Vachani, Anil; Snyder, Nathaniel W

2016-12-01

A multiplexed quantitative method for the analysis of three major unconjugated steroids in human serum by stable isotope dilution liquid chromatography-high resolution mass spectrometry (LC-HRMS) was developed and validated on a Q Exactive Plus hybrid quadrupole/Orbitrap mass spectrometer. This quantification utilized isotope dilution and Girard P derivatization on the keto-groups of testosterone (T), androstenedione (AD) and dehydroepiandrosterone (DHEA) to improve ionization efficiency using electrospray ionization. Major isomeric compounds to T and DHEA; the inactive epimer of testosterone (epiT), and the metabolite of AD, 5α-androstanedione (5α-AD) were completely resolved on a biphenyl column within an 18min method. Inter- and intra-day method validation using LC-HRMS with qualifying product ions was performed and acceptable analytical performance was achieved. The method was further validated by comparing steroid levels from 100μL of serum from young vs older subjects. Since this approach provides high-dimensional HRMS data, untargeted analysis by age group was performed. DHEA and T were detected among the top analytes most significantly different across the two groups after untargeted LC-HRMS analysis, as well as a number of other still unknown metabolites, indicating the potential for combined targeted/untargeted analysis in steroid analysis. Copyright © 2016 Elsevier Inc. All rights reserved.

Effects of past and recent blood pressure and cholesterol level on coronary heart disease and stroke mortality, accounting for measurement error.

PubMed

Boshuizen, Hendriek C; Lanti, Mariapaola; Menotti, Alessandro; Moschandreas, Joanna; Tolonen, Hanna; Nissinen, Aulikki; Nedeljkovic, Srecko; Kafatos, Anthony; Kromhout, Daan

2007-02-15

The authors aimed to quantify the effects of current systolic blood pressure (SBP) and serum total cholesterol on the risk of mortality in comparison with SBP or serum cholesterol 25 years previously, taking measurement error into account. The authors reanalyzed 35-year follow-up data on mortality due to coronary heart disease and stroke among subjects aged 65 years or more from nine cohorts of the Seven Countries Study. The two-step method of Tsiatis et al. (J Am Stat Assoc 1995;90:27-37) was used to adjust for regression dilution bias, and results were compared with those obtained using more commonly applied methods of adjustment for regression dilution bias. It was found that the commonly used univariate adjustment for regression dilution bias overestimates the effects of both SBP and cholesterol compared with multivariate methods. Also, the two-step method makes better use of the information available, resulting in smaller confidence intervals. Results comparing recent and past exposure indicated that past SBP is more important than recent SBP in terms of its effect on coronary heart disease mortality, while both recent and past values seem to be important for effects of cholesterol on coronary heart disease mortality and effects of SBP on stroke mortality. Associations between serum cholesterol concentration and risk of stroke mortality are weak.

STRUCTURAL, SURFACE MORPHOLOGICAL AND MAGNETIC STUDIES OF Zn1-xFexS (x=0.00-0.10) DILUTED MAGNETIC SEMICONDUCTORS GROWN BY CO-PRECIPITATION METHOD

NASA Astrophysics Data System (ADS)

Hassan, M.; Ghazanfar, M.; Arooj, N.; Riaz, S.; Hussain, S. Sajjad; Naseem, S.

We have fabricated Zn1-xFexS (x=0.00, 0.02, 0.04, 0.06, 0.08 and 0.10) diluted magnetic semiconductors using co-precipitation method. X-ray diffraction patterns depict that Zn1-xFexS appears as a dominant phase with cubic zinc blende structure and nanoscale crystallite size. In addition, a secondary phase of rhombohedral ZnS also appears; however, no additional phase arises that primarily belongs to Fe dopant. Using Debye-Scherrer relation, the crystallite size is found to be in the range of 20-27nm, which is in good agreement with the crystallite size calculated using the Williamson-Hall (WH) plot method. The appearance of secondary phase provoked to study the residual strain using Stokes-Wilson equation, which is nearly consistent to that observed using WH plot method. The surface morphology, revealed using scanning electron microscopy, depicts non-uniform surface structure with a variety of grains and void dimensions. Hysteresis loops measured for Zn1-xFexS at room temperature (RT) illustrate a paramagnetic behavior at higher fields; however, small ferromagnetic behavior is evident due to the small openings of the measured hysteresis loops around the origin. The measured RT ferromagnetism reveals the potential spintronic device applications of the studied diluted magnetic semiconductors.

The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques.

PubMed Central

Sabattini, E; Bisgaard, K; Ascani, S; Poggi, S; Piccioli, M; Ceccarelli, C; Pieri, F; Fraternali-Orcioni, G; Pileri, S A

1998-01-01

AIM: To assess a newly developed immunohistochemical detection system, the EnVision++. METHODS: A large series of differently processed normal and pathological samples and 53 relevant monoclonal antibodies were chosen. A chessboard titration assay was used to compare the results provided by the EnVision++ system with those of the APAAP, CSA, LSAB, SABC, and ChemMate methods, when applied either manually or in a TechMate 500 immunostainer. RESULTS: With the vast majority of the antibodies, EnVision++ allowed two- to fivefold higher dilutions than the APAAP, LSAB, SABC, and ChemMate techniques, the staining intensity and percentage of expected positive cells being the same. With some critical antibodies (such as the anti-CD5), it turned out to be superior in that it achieved consistently reproducible results with differently fixed or overfixed samples. Only the CSA method, which includes tyramide based enhancement, allowed the same dilutions as the EnVision++ system, and in one instance (with the anti-cyclin D1 antibody) represented the gold standard. CONCLUSIONS: The EnVision++ is an easy to use system, which avoids the possibility of disturbing endogenous biotin and lowers the cost per test by increasing the dilutions of the primary antibodies. Being a two step procedure, it reduces both the assay time and the workload. Images PMID:9797726

Successful use of the Cryolock device for cryopreservation of scarce human ejaculate and testicular spermatozoa.

PubMed

Stein, A; Shufaro, Y; Hadar, S; Fisch, B; Pinkas, H

2015-03-01

The existing methods for cryopreservation of very low count sperm samples are complex and sub-optimal for individual spermatozoa. Our purpose is to establish an effective simple method for cryoprotecting individual spermatozoa. Samples from patients with OTA were mixed with TYB or HEPES-buffered salt solution with glycerol + glucose and placed on a Cryolock that was plunged directly into liquid nitrogen or exposed to its vapors. Thawing was performed by direct immersion into a drop of warmed medium. The favorable method was tested on diluted samples (10-50 cells) and leftover TESE specimens from patients with azoospermia. Cryopreservation was considered successful if >30 spermatozoa, (>3 motile), or >5 spermatozoa (>1 motile) in diluted and TESE samples, were detected post-thawing. A significantly higher survival rate of seminal spermatozoa was obtained when using the Cryolock with TYB and freezing with liquid nitrogen vapor, compared to HEPES glycerol-glucose (95 vs. 35% respectively). Plunging the Cryolock into liquid nitrogen was detrimental. Cryolock combined with TYB cryoprotection and liquid nitrogen vapor freezing was highly effective for cryopreservation of individual spermatozoa in diluted and TESE samples. The Cryolock may serve for freezing very low-count sperm samples and individual spermatozoa. This method offers simplicity, efficacy, use of available materials, without requiring micromanipulation equipment or skills. © 2015 American Society of Andrology and European Academy of Andrology.

An Effective CB Material from Combined Components of Triosyn(Trademark) Resin and Surface Enhanced Carbon

DTIC Science & Technology

2002-01-01

and water solubility, as well as low toxicity and the ability to be easily detected by analytical methods. It is the approved simulant used in CRDC... coliphage (ATCC 15597-B1), a bacterial virus known for its survival capacities in the environment, was used as a biological agent to challenge the...buffer (NDS) kept in a 35°C water bath until complete dissolution of the membranes. Dilutions of MS2 assays were made in sterile PBS. Serial dilutions of

Automatic tracer-dilution method used for stage-discharge ratings and streamflow hydrographs on small Iowa streams

USGS Publications Warehouse

Soenksen, P.J.

1990-01-01

Tracer-dilution discharge measurements were made during 14 flow periods at six stations from 1986 through 1988 water years. Ratings were developed at three stations with the aid of these measurements. A loop rating was identified at one station during rapidly-changing flow conditions. Incomplete mixing and dye loss to sediment apparently were problems at some stations. Stage hydrographs were recorded for 38 flows at seven stations. Limited data on background fluorescence during high flows were also obtained.

Dispersion of nanosized ceramic powders in aqueous suspensions

NASA Astrophysics Data System (ADS)

Chera, L.; Palcevskis, E.; Berzins, M.; Lipe, A.; Jansone, I.

2007-12-01

Seven commercially available dispersants have been applied to produce high concentrated aqueous suspensions of the nanosized alumina and partially stabilized zirconia powders processed by the plasma technique. Simultaneously, the electrokinetic behaviour of powders has been investigated in diluted suspensions by microelectrophoresis method. Zeta potential measurements are used to estimate the influence of selected dispersants on the electrokinetic properties of the powder surface. On the basis of obtained data the correlation between the surface electrokinetic properties in dilute suspensions and reached maximal suspension concentration is discussed.

Rotor for processing liquids using movable capillary tubes

DOEpatents

Johnson, W.F.; Burtis, C.A.; Walker, W.A.

1987-07-17

A rotor assembly for processing liquids, especially whole blood samples, is disclosed. The assembly includes apparatus for separating non-liquid components of whole blood samples from liquid components, apparatus for diluting the separated liquid component with a diluent and apparatus for transferring the diluted sample to an external apparatus for analysis. The rotor assembly employs several movable capillary tubes to handle the sample and diluents. A method for using the rotor assembly to process liquids is also described. 5 figs.

[Rapid bioluminescent antibiotic susceptibility assay].

PubMed

Frundzhian, V G; Ugarova, N N; Blatun, L A; Terekhova, R P; Rusanova, E V

2009-01-01

Rapid testing of pathogen susceptibility to antibiotics is of great practical value for rational chemotherapy of pyoinflammatory deseases and postoperative complications of microbial etiology. The standard microbiological methods, i.e., the disk diffusion method and the method of serial dilutions are labour- and time-consuming (not less than 18-36 hours). The method of the authors is based on measuring bioluminescence resulting from interaction of adenosine-5'-triphosphate (ATP) and ATP reagent, a standard reaction mixture of firefly luciferase (an enzyme) and luciferin. The bioluminescence intensity is proportional to the ATP concentration in the reaction mixture and the ATP concentration is proportional to the number of the pathogen viable cells in the sample. The bioluminescence intensity value in the pathogen suspension aliquots with and without (control) the antibiotic were compared after the incubation for 5 hours and the coefficient of the microbial cell growth inhibition was calculated. Satisfactory correlation (R2 > 88%) of the results of the bioluminescent assay and the assay with the disk diffusion method and the method of serial dilutions was observed.

Modifications to the NIST reference measurement procedure (RMP) for the determination of serum glucose by isotope dilution gas chromatography/mass spectrometry.

PubMed

Prendergast, Jocelyn L; Sniegoski, Lorna T; Welch, Michael J; Phinney, Karen W

2010-07-01

The definitive method (DM), now known as the reference measurement procedure (RMP), for the analysis of glucose in serum was originally published in 1982 by the National Institute of Standards and Technology (NIST). Over the years the method has been subject to a number of modifications to adapt to newer technologies and simplify sample preparation. We discuss here an adaptation of the method associated with serum glucose measurements using a modified isotope dilution gas chromatography/mass spectrometry (ID-GC/MS) method. NIST has used this modified method to certify the concentrations of glucose in SRM 965b, Glucose in Frozen Human Serum, and SRM 1950, Metabolites in Human Plasma. Comparison of results from the revised method with certified values for existing Standard Reference Materials (SRMs) demonstrated that these modifications have not affected the quality of the measurements, giving both good precision and accuracy, while reducing the sample preparation time by a day and a half.

Improvement of the energy conversion efficiency of Chlorella pyrenoidosa biomass by a three-stage process comprising dark fermentation, photofermentation, and methanogenesis.

PubMed

Xia, Ao; Cheng, Jun; Ding, Lingkan; Lin, Richen; Huang, Rui; Zhou, Junhu; Cen, Kefa

2013-10-01

The effects of pre-treatment methods on saccharification and hydrogen fermentation of Chlorella pyrenoidosa biomass were investigated. When raw biomass and biomass pre-treated by steam heating, by microwave heating, and by ultrasonication were used as feedstock, the hydrogen yields were only 8.8-12.7 ml/g total volatile solids (TVS) during dark fermentation. When biomass was pre-treated by steam heating with diluted acid and by microwave heating with diluted acid, the dark hydrogen yields significantly increased to 75.6 ml/g TVS and 83.3 ml/g TVS, respectively. Steam heating with diluted acid is the preferred pre-treatment method of C. pyrenoidosa biomass to improve hydrogen yield during dark fermentation and photofermentation, which is followed by methanogenesis to increase energy conversion efficiency (ECE). A total hydrogen yield of 198.3 ml/g TVS and a methane yield of 186.2 ml/g TVS corresponding to an overall ECE of 34.0% were obtained through the three-stage process (dark fermentation, photofermentation, and methanogenesis). Copyright © 2013 Elsevier Ltd. All rights reserved.

Reductive amination derivatization for the quantification of garlic components by isotope dilution analysis.

PubMed

Lin, Yi-Reng; Huang, Mei-Fang; Wu, You-Ying; Liu, Meng-Chieh; Huang, Jing-Heng; Chen, Ziyu; Shiue, Yow-Ling; Wu, Chia-En; Liang, Shih-Shin

2017-09-01

In this work, we synthesized internal standards for four garlic organosulfur compounds (OSCs) by reductive amination with 13 C, D 2 -formaldehyde, and developed an isotope dilution analysis method to quantitate these organosulfur components in garlic samples. Internal standards were synthesized for internal absolute quantification of S-allylcysteine (SAC), S-allylcysteine sulfoxide (alliin), S-methylcysteine (SMC), and S-ethylcysteine (SEC). We used a multiple reaction monitoring (MRM) to detect 13 C, D 2 -formaldehyde-modified OSCs by ultrahigh-performance liquid phase chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) and obtained MS spectra showing different ratios of 13 C, D 2 -formaldehyde-modified and H 2 -formaldehyde-modified compounds. The resulting labeled and unlabeled OSCs were exhibited correlation coefficient (R 2 ) ranged from 0.9989 to 0.9994, respectively. The average recoveries for four OSCs at three concentration levels ranged from 89% to 105%. By 13 C, D 2 -formaldehyde and sodium cyanoborohydride, the reductive amination-based method can be utilized to generate novel internal standard for isotope dilution and to extend the quantitative application. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cloning of Plasmodium falciparum by single-cell sorting

PubMed Central

Miao, Jun; Li, Xiaolian; Cui, Liwang

2010-01-01

Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples. PMID:20435038

Standardized methods and quality control limits for agar and broth microdilution susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum.

PubMed

Waites, Ken B; Duffy, Lynn B; Bébéar, Cécile M; Matlow, Anne; Talkington, Deborah F; Kenny, George E; Totten, Patricia A; Bade, Donald J; Zheng, Xiaotian; Davidson, Maureen K; Shortridge, Virginia D; Watts, Jeffrey L; Brown, Steven D

2012-11-01

An international multilaboratory collaborative study was conducted to develop standard media and consensus methods for the performance and quality control of antimicrobial susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum using broth microdilution and agar dilution techniques. A reference strain from the American Type Culture Collection was designated for each species, which was to be used for quality control purposes. Repeat testing of replicate samples of each reference strain by participating laboratories utilizing both methods and different lots of media enabled a 3- to 4-dilution MIC range to be established for drugs in several different classes, including tetracyclines, macrolides, ketolides, lincosamides, and fluoroquinolones. This represents the first multilaboratory collaboration to standardize susceptibility testing methods and to designate quality control parameters to ensure accurate and reliable assay results for mycoplasmas and ureaplasmas that infect humans.

SLDAssay: A software package and web tool for analyzing limiting dilution assays.

PubMed

Trumble, Ilana M; Allmon, Andrew G; Archin, Nancie M; Rigdon, Joseph; Francis, Owen; Baldoni, Pedro L; Hudgens, Michael G

2017-11-01

Serial limiting dilution (SLD) assays are used in many areas of infectious disease related research. This paper presents SLDAssay, a free and publicly available R software package and web tool for analyzing data from SLD assays. SLDAssay computes the maximum likelihood estimate (MLE) for the concentration of target cells, with corresponding exact and asymptotic confidence intervals. Exact and asymptotic goodness of fit p-values, and a bias-corrected (BC) MLE are also provided. No other publicly available software currently implements the BC MLE or the exact methods. For validation of SLDAssay, results from Myers et al. (1994) are replicated. Simulations demonstrate the BC MLE is less biased than the MLE. Additionally, simulations demonstrate that exact methods tend to give better confidence interval coverage and goodness-of-fit tests with lower type I error than the asymptotic methods. Additional advantages of using exact methods are also discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

Rifaximin-resistant Clostridium difficile strains isolated from symptomatic patients.

PubMed

Reigadas, E; Muñoz-Pacheco, P; Vázquez-Cuesta, S; Alcalá, L; Marín, M; Martin, A; Bouza, E

2017-12-01

Rifaximin has been proposed as an alternative treatment for specific cases of Clostridium difficile infection (CDI) and intestinal decontamination. Rifaximin-resistant C. difficile has occasionally been reported. Antibiotic susceptibility testing relies on anaerobic agar dilution (reference method), which is cumbersome and not routinely used. There is no commercial test for detection of resistance to rifaximin. To assess resistance to rifaximin by C. difficile and to evaluate the correlation between the results of the rifampicin E-test and susceptibility to rifaximin. We compared the in vitro susceptibility of clinical CDI isolates to rifaximin over a 6-month period using the agar dilution method with susceptibility to rifampicin using the E-test. All isolates were characterized using PCR-ribotyping. Clinical data were recorded prospectively. We recovered 276 consecutive C. difficile isolates and found that 32.2% of episodes were caused by rifaximin-resistant strains. The MICs for rifaximin ranged from <0.0009-256 mg/L, with a geometric mean (GM) of 0.256 mg/L, an MIC 50/90 of 0.015/>256 mg/L. Rifaximin and rifampicin MICs were comparable, and all strains classed as resistant by agar dilution were correctly classified as resistant by E-test. The most common ribotypes were 001 (37.2%), 078/126 (14.3%), and 014 (12.0%). Ribotype 001 exhibited the highest MICs for rifaximin. Resistance to rifaximin was common; resistance rates were higher in ribotype 001 strains. Susceptibility to rifaximin determined by agar dilution correlated with susceptibility to rifampicin determined using the E-test, including rifaximin-resistant strains. Our results suggest that the rifampicin E-test is a valid method for the prediction of rifaximin-resistant C. difficile. Copyright © 2017 Elsevier Ltd. All rights reserved.

Evaluation of analytical methodology for the detection of hormones and their attenuation during aquifer recharge and recovery cycles.

PubMed

de Lima Stebbins, Daniela; Docs, Jon; Lowe, Paula; Cohen, Jason; Lei, Hongxia

2016-05-18

The hormones listed in the screening survey list 2 of the Unregulated Contaminant Monitoring Rule 3 (estrone, 17-β-estradiol, 17-α-ethynylestradiol, 16-α-hydroxyestradiol (estriol), equilin, testosterone and 4-androstene-3,17-dione) were analyzed by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Two analytical methods were compared: EPA method 539 and the isotope dilution method. EPA method 539 was successfully utilized in river and drinking water matrices with fortified recoveries of 98.9 to 108.5%. Samples from the Hillsborough River reflected levels below the method detection limit (MDL) for the majority of the analytes, except estrone (E1), which was detected at very low concentrations (<0.5 to 1 ng L(-1)) in the majority of samples. No hormones were detected in drinking water samples. The isotope dilution method was used to analyze reclaimed and aquifer storage and recovery (ASR) water samples as a result of strong matrix/solid phase extraction (SPE) losses observed in these more complex matrices. Most of the compounds were not detected or found at relatively low concentrations in the ASR samples. Attenuation of 50 to 99.1% was observed as a result of the ASR recharge/recovery cycles for most of the hormones, except for estriol (E3). Relatively stable concentrations of E3 were found, with only 10% attenuation at one of the sites and no measureable attenuation at another location. These results have substantiated that while EPA method 539 works well for most environmental samples, the isotope dilution method is more robust when dealing with complex matrices such as reclaimed and ASR samples.

Determination of low methylmercury concentrations in peat soil samples by isotope dilution GC-ICP-MS using distillation and solvent extraction methods.

PubMed

Pietilä, Heidi; Perämäki, Paavo; Piispanen, Juha; Starr, Mike; Nieminen, Tiina; Kantola, Marjatta; Ukonmaanaho, Liisa

2015-04-01

Most often, only total mercury concentrations in soil samples are determined in environmental studies. However, the determination of extremely toxic methylmercury (MeHg) in addition to the total mercury is critical to understand the biogeochemistry of mercury in the environment. In this study, N2-assisted distillation and acidic KBr/CuSO4 solvent extraction methods were applied to isolate MeHg from wet peat soil samples collected from boreal forest catchments. Determination of MeHg was performed using a purge and trap GC-ICP-MS technique with a species-specific isotope dilution quantification. Distillation is known to be more prone to artificial MeHg formation compared to solvent extraction which may result in the erroneous MeHg results, especially with samples containing high amounts of inorganic mercury. However, methylation of inorganic mercury during the distillation step had no effect on the reliability of the final MeHg results when natural peat soil samples were distilled. MeHg concentrations determined in peat soil samples after distillation were compared to those determined after the solvent extraction method. MeHg concentrations in peat soil samples varied from 0.8 to 18 μg kg(-1) (dry weight) and the results obtained with the two different methods did not differ significantly (p=0.05). The distillation method with an isotope dilution GC-ICP-MS was shown to be a reliable method for the determination of low MeHg concentrations in unpolluted soil samples. Furthermore, the distillation method is solvent-free and less time-consuming and labor-intensive when compared to the solvent extraction method. Copyright © 2014 Elsevier Ltd. All rights reserved.

Probing critical behavior of 2D Ising ferromagnet with diluted bonds using Wang-Landau algorithm

NASA Astrophysics Data System (ADS)

Ridha, N. A.; Mustamin, M. F.; Surungan, T.

2018-03-01

Randomness is an important subject in the study of phase transition as defect and impurity may present in any real material. The pre-existing ordered phase of a pure system can be affected or even ruined by the presence of randomness. Here we study ferromagnetic Ising model on a square lattice with a presence of randomness in the form of bond dilution. The pure system of this model is known to experience second order phase transition, separating between the high temperature paramagnetic and low-temperature ferromagnetic phase. We used Wang-Landau algorithm of Monte Carlo method to obtain the density of states from which we extract the ensemble average of energy and the specific heat. We observed the signature of phase transition indicated by the diverging peak of the specific heat as system sizes increase. These peaks shift to the lower temperature side as the dilution increases. The lower temperature ordered phase preserves up to certain concentration of dilution and is totally ruined when the bonds no longer percolates.

Infinitely Dilute Partial Molar Properties of Proteins from Computer Simulation

PubMed Central

2015-01-01

A detailed understanding of temperature and pressure effects on an infinitely dilute protein’s conformational equilibrium requires knowledge of the corresponding infinitely dilute partial molar properties. Established molecular dynamics methodologies generally have not provided a way to calculate these properties without either a loss of thermodynamic rigor, the introduction of nonunique parameters, or a loss of information about which solute conformations specifically contributed to the output values. Here we implement a simple method that is thermodynamically rigorous and possesses none of the above disadvantages, and we report on the method’s feasibility and computational demands. We calculate infinitely dilute partial molar properties for two proteins and attempt to distinguish the thermodynamic differences between a native and a denatured conformation of a designed miniprotein. We conclude that simple ensemble average properties can be calculated with very reasonable amounts of computational power. In contrast, properties corresponding to fluctuating quantities are computationally demanding to calculate precisely, although they can be obtained more easily by following the temperature and/or pressure dependence of the corresponding ensemble averages. PMID:25325571

Evaluating the quality of a cell counting measurement process via a dilution series experimental design.

PubMed

Sarkar, Sumona; Lund, Steven P; Vyzasatya, Ravi; Vanguri, Padmavathy; Elliott, John T; Plant, Anne L; Lin-Gibson, Sheng

2017-12-01

Cell counting measurements are critical in the research, development and manufacturing of cell-based products, yet determining cell quantity with accuracy and precision remains a challenge. Validating and evaluating a cell counting measurement process can be difficult because of the lack of appropriate reference material. Here we describe an experimental design and statistical analysis approach to evaluate the quality of a cell counting measurement process in the absence of appropriate reference materials or reference methods. The experimental design is based on a dilution series study with replicate samples and observations as well as measurement process controls. The statistical analysis evaluates the precision and proportionality of the cell counting measurement process and can be used to compare the quality of two or more counting methods. As an illustration of this approach, cell counting measurement processes (automated and manual methods) were compared for a human mesenchymal stromal cell (hMSC) preparation. For the hMSC preparation investigated, results indicated that the automated method performed better than the manual counting methods in terms of precision and proportionality. By conducting well controlled dilution series experimental designs coupled with appropriate statistical analysis, quantitative indicators of repeatability and proportionality can be calculated to provide an assessment of cell counting measurement quality. This approach does not rely on the use of a reference material or comparison to "gold standard" methods known to have limited assurance of accuracy and precision. The approach presented here may help the selection, optimization, and/or validation of a cell counting measurement process. Published by Elsevier Inc.

Quantifying Trophic and Demographic Rates of Plankton-Rich Layers in East Sound, Orcas Island, Washington

DTIC Science & Technology

2007-09-30

different methods are used, two independent methods to measure primary productivity and the dilution method to measure heterotrophic protists grazing...relationships for dinoflagellates, diatoms, and other protist plankton. Limnology and Oceanography 45: 569-579 Menden-Deuer S, EJ Lessard & J

21 CFR 177.1960 - Vinyl chloride-hexene-1 copolymers.

Code of Federal Regulations, 2013 CFR

2013-04-01

... accepted applicability. (ii) Inherent viscosity in cyclohexanone at 30 °C is not less than 0.59 deciliters per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution Viscosity of...

21 CFR 177.1960 - Vinyl chloride-hexene-1 copolymers.

Code of Federal Regulations, 2012 CFR

2012-04-01

... accepted applicability. (ii) Inherent viscosity in cyclohexanone at 30 °C is not less than 0.59 deciliters per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution Viscosity of...

21 CFR 177.1960 - Vinyl chloride-hexene-1 copolymers.

Code of Federal Regulations, 2011 CFR

2011-04-01

... accepted applicability. (ii) Inherent viscosity in cyclohexanone at 30 °C is not less than 0.59 deciliters per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution Viscosity of...

Screening methodology for needs of roadway lighting.

DOT National Transportation Integrated Search

2003-01-01

Screening methods of AASHTO and NCHRP that assess the local potential for fixed roadway lighting to decrease nighttime crashes have not been updated since the 1970's. The methods dilute the influence of important factors, are inadequate for locations...

76 FR 63615 - Environmental Science Center Microbiology Laboratory; Notice of Public Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-13

...-dilution method (UDM) and the status and implementation of a new test method, the Organization for Economic... using the method for registration purposes, the method is currently in use as part of the post market... associated with the S. aureus and P. aeruginosa methodologies, which are designed to further optimize and...

Validation of an isotope dilution, ICP-MS method based on internal mass bias correction for the determination of trace concentrations of Hg in sediment cores.

PubMed

Ciceri, E; Recchia, S; Dossi, C; Yang, L; Sturgeon, R E

2008-01-15

The development and validation of a method for the determination of mercury in sediments using a sector field inductively coupled plasma mass spectrometer (SF-ICP-MS) for detection is described. The utilization of isotope dilution (ID) calibration is shown to solve analytical problems related to matrix composition. Mass bias is corrected using an internal mass bias correction technique, validated against the traditional standard bracketing method. The overall analytical protocol is validated against NRCC PACS-2 marine sediment CRM. The estimated limit of detection is 12ng/g. The proposed procedure was applied to the analysis of a real sediment core sampled to a depth of 160m in Lake Como, where Hg concentrations ranged from 66 to 750ng/g.

Effect of laser power on clad metal in laser-TIG combined metal cladding

NASA Astrophysics Data System (ADS)

Utsumi, Akihiro; Hino, Takanori; Matsuda, Jun; Tasoda, Takashi; Yoneda, Masafumi; Katsumura, Munehide; Yano, Tetsuo; Araki, Takao

2003-03-01

TIG arc welding has been used to date as a method for clad welding of white metal as bearing material. We propose a new clad welding process that combines a CO2 laser and a TIG arc, as a method for cladding at high speed. We hypothesized that this method would permit appropriate control of the melted quantity of base metal by varying the laser power. We carried out cladding while varying the laser power, and investigated the structure near the boundary between the clad layer and the base metal. Using the laser-TIG combined cladding, we found we were able to control appropriately the degree of dilution with the base metal. By applying this result to subsequent cladding, we were able to obtain a clad layer of high quality, which was slightly diluted with the base metal.

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.

Biomass treatment method

DOEpatents

Friend, Julie; Elander, Richard T.; Tucker, III; Melvin P.; Lyons, Robert C.

2010-10-26

A method for treating biomass was developed that uses an apparatus which moves a biomass and dilute aqueous ammonia mixture through reaction chambers without compaction. The apparatus moves the biomass using a non-compressing piston. The resulting treated biomass is saccharified to produce fermentable sugars.

Lab tests on the biodegradation of chemically dispersed oil should consider the rapid dilution that occurs at sea.

PubMed

Lee, Kenneth; Nedwed, Tim; Prince, Roger C; Palandro, David

2013-08-15

Most crude oils spread on open water to an average thickness as low as 0.1 mm. The application of dispersants enhances the transport of oil as small droplets into the water column, and when combined with the turbulence of 1 m waves will quickly entrain oil into the top 1 m of the water column, where it rapidly dilutes to concentrations less than 100 ppm. In less than 24 h, the dispersed oil is expected to mix into the top 10 m of the water column and be diluted to concentrations well below 10 ppm, with dilution continuing as time proceeds. Over the multiple weeks that biodegradation takes place, dispersed oil concentrations are expected to be below 1 ppm. Measurements from spills and wave basin studies support these calculations. Published laboratory studies focused on the quantification of contaminant biodegradation rates have used concentrations orders of magnitude greater than this, as it was necessary to ensure the concentrations of hydrocarbons and other chemicals were higher than the detection limits of chemical analysis. However, current analytical methods can quantify individual alkanes and PAHs (and their alkyl homologues) at ppb and ppm levels. To simulate marine biodegradation of dispersed oil at dilute concentrations commonly encountered in the field, laboratory studies should be conducted at similarly low hydrocarbon concentrations. Copyright © 2013 Elsevier Ltd. All rights reserved.

Determination of 232Th in urine by ICP-MS for individual monitoring purposes.

PubMed

Baglan, N; Cossonnet, C; Ritt, J

2001-07-01

Thorium is naturally occurring in various ores used for industrial purposes and has numerous applications. This paper sets out to investigate urine analysis as a suitable monitoring approach for workers potentially exposed to thorium. Due to its biokinetic behavior and its low solubility, urinary concentrations are generally very low, requiring therefore high sensitivity analytical methods. An analytical procedure has been developed for detecting 232Th concentrations of below 1 mBq L(-1) quickly and easily. Due to the long half-life (1.41 x 10(10) y) of 232Th, the potential of a procedure based on urine sample dilution and ICP-MS (inductively coupled plasma-mass spectrometry) measurement was investigated first. Two dilution factors were chosen: 100, which is more suitable for long-term measurement trials, and 20, which increases sensitivity. It has been shown that a 100-fold dilution can be used to measure concentrations of below 1 mBq L(-1), whereas a 20-fold one can be used to reach concentrations of below 0.06 mBq L(-1). Then, on the basis of the limitation of the procedure based on urine dilution, the suitable field of application for the different procedures (100-fold and 20-fold dilution and also a chemical purification followed by an ICP-MS measurement) was determined in relation to monitoring objectives.

Wheat and ultra high diluted gibberellic acid--further experiments and re-analysis of data.

PubMed

Endler, Peter Christian; Scherer-Pongratz, Waltraud; Lothaller, Harald; Stephen, Saundra

2015-10-01

Following studies (a) on wheat seedlings and ultra high diluted silver nitrate, and (b) on amphibians and an ultra high diluted hormone, (c) a bio-assay on wheat and extremely diluted gibberellic acid was standardized. This assay was intended to combine the easy-to-handle aspect of (a) and biologically interesting aspects of (b). The purpose of the data analysis presented here was to investigate the influence of an extreme dilution of gibberellic acid on wheat stalk length and to determine the influence of external factors on the experimental outcome. Grains of winter wheat (Triticum aestivum, Capo variety) were observed under the influence of extremely diluted gibberellic acid (10(-30)) prepared by stepwise dilution and agitation according to a protocol derived from homeopathy ('G30x'). Analogously prepared water was used for control ('W30x'). 16 experiments including 8000+8000 grains were performed by 9 researchers. Experiments that were performed between January and April showed inconsistent results, whereas most of the experiments performed between September and December showed shorter stalks in the G30x group. This was confirmed by correlation analysis (p<0.01). Thus winter/spring experiments and autumn experiments were analysed separately. When all 10 autumn experiments were pooled, mean stalk lengths (mm) were 48.3±21.4 for the verum group and 52.1±20.4 for control (mean±SD) at grain level (N=5000 per group) and ±5.3 and ±5.1 respectively at dish level. In other words, verum stalk length (92.67%) was 7.33% smaller than control stalk length (100%). The effect size is small when calculation is done on the basis of grains (d=0.18) but, due to the smaller SD at dish level, medium when done on the basis of dishes (d=0.73). The inhibiting effect was observed by 6 of the 6 researchers who performed the autumn experiments. The model may be useful for further research as there exists a theoretical justification due to previous studies with wheat and extremely diluted silver nitrate, as well as to previous studies with amphibians and diluted hormones, and its methods are well standardized. Data confirm the hypothesis that information can be stored in the test liquid, even at a dilution of the original substance beyond Avogadro's value; and that the wheat bio-assay is sensitive to such information. Copyright © 2015 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.

Guideline on Isotope Dilution Mass Spectrometry

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

Gaffney, Amy

Isotope dilution mass spectrometry is used to determine the concentration of an element of interest in a bulk sample. It is a destructive analysis technique that is applicable to a wide range of analytes and bulk sample types. With this method, a known amount of a rare isotope, or ‘spike’, of the element of interest is added to a known amount of sample. The element of interest is chemically purified from the bulk sample, the isotope ratio of the spiked sample is measured by mass spectrometry, and the concentration of the element of interest is calculated from this result. Thismore » method is widely used, although a mass spectrometer required for this analysis may be fairly expensive.« less

Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction.

PubMed

Mohammadi, Samira; Esfahani, Bahram Nasr; Moghim, Sharareh; Mirhendi, Hossein; Zaniani, Fatemeh Riyahi; Safaei, Hajieh Ghasemian; Fazeli, Hossein; Salehi, Mahshid

2017-01-01

Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA ® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples. The CTAB method showed more positive results at 1:10-1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor.

Development of PMA real-time PCR method to quantify viable cells of Pantoea agglomerans CPA-2, an antagonist to control the major postharvest diseases on oranges.

PubMed

Soto-Muñoz, Lourdes; Teixidó, Neus; Usall, Josep; Viñas, Inmaculada; Crespo-Sempere, Ana; Torres, Rosario

2014-06-16

Dilution plating is the quantification method commonly used to estimate the population level of postharvest biocontrol agents, but this method does not permit a distinction among introduced and indigenous strains. Recently, molecular techniques based on DNA amplification such as quantitative real-time PCR (qPCR) have been successfully applied for their high strain-specific detection level. However, the ability of qPCR to distinguish viable and nonviable cells is limited. A promising strategy to avoid this issue relies on the use of nucleic acid intercalating dyes, such as propidium monoazide (PMA), as a sample pretreatment prior to the qPCR. The objective of this study was to optimize a protocol based on PMA pre-treatment samples combined with qPCR to distinguish and quantify viable cells of the biocontrol agent P. agglomerans CPA-2 applied as a postharvest treatment on orange. The efficiency of PMA-qPCR method under the established conditions (30μM PMA for 20min of incubation followed by 30min of LED light exposure) was evaluated on an orange matrix. Results showed no difference in CFU or cells counts of viable cells between PMA-qPCR and dilution plating. Samples of orange matrix inoculated with a mixture of viable/dead cells showed 5.59log10 CFU/ml by dilution plating, 8.25log10 cells/ml by qPCR, and 5.93log10 cells/ml by PMA-qPCR. Furthermore, samples inoculated with heat-killed cells were not detected by dilution plating and PMA-qPCR, while by qPCR was of 8.16log10 cells/ml. The difference in quantification cycles (Cq) among qPCR and PMA-qPCR was approximately 16cycles, which means a reduction of 65,536 fold of the dead cells detected. In conclusion, PMA-qPCR method is a suitable tool for quantify viable CPA-2 cells, which could be useful to estimate the ability of this antagonist to colonize the orange surface. Copyright © 2014 Elsevier B.V. All rights reserved.

Cluster Dynamics Modeling with Bubble Nucleation, Growth and Coalescence

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

de Almeida, Valmor F.; Blondel, Sophie; Bernholdt, David E.

The topic of this communication pertains to defect formation in irradiated solids such as plasma-facing tungsten submitted to helium implantation in fusion reactor com- ponents, and nuclear fuel (metal and oxides) submitted to volatile ssion product generation in nuclear reactors. The purpose of this progress report is to describe ef- forts towards addressing the prediction of long-time evolution of defects via continuum cluster dynamics simulation. The di culties are twofold. First, realistic, long-time dynamics in reactor conditions leads to a non-dilute di usion regime which is not accommodated by the prevailing dilute, stressless cluster dynamics theory. Second, long-time dynamics callsmore » for a large set of species (ideally an in nite set) to capture all possible emerging defects, and this represents a computational bottleneck. Extensions beyond the dilute limit is a signi cant undertaking since no model has been advanced to extend cluster dynamics to non-dilute, deformable conditions. Here our proposed approach to model the non-dilute limit is to monitor the appearance of a spatially localized void volume fraction in the solid matrix with a bell shape pro le and insert an explicit geometrical bubble onto the support of the bell function. The newly cre- ated internal moving boundary provides the means to account for the interfacial ux of mobile species into the bubble, and the growth of bubbles allows for coalescence phenomena which captures highly non-dilute interactions. We present a preliminary interfacial kinematic model with associated interfacial di usion transport to follow the evolution of the bubble in any number of spatial dimensions and any number of bubbles, which can be further extended to include a deformation theory. Finally we comment on a computational front-tracking method to be used in conjunction with conventional cluster dynamics simulations in the non-dilute model proposed.« less

Short communication: milk output in llamas (Lama glama) in relation to energy intake and water turnover measured by an isotope dilution technique.

PubMed

Riek, A; Klinkert, A; Gerken, M; Hummel, J; Moors, E; Südekum, K-H

2013-03-01

Despite the fact that llamas have become increasingly popular as companion and farm animals in both Europe and North America, scientific knowledge on their nutrient requirements is scarce. Compared with other livestock species, relatively little is known especially about the nutrient and energy requirements for lactating llamas. Therefore, we aimed to measure milk output in llama dams using an isotope dilution technique and relate it to energy intakes at different stages of lactation. We also validated the dilution technique by measuring total water turnover (TWT) directly and comparing it with values estimated by the isotope dilution technique. Our study involved 5 lactating llama dams and their suckling young. Milk output and TWT were measured at 4 stages of lactation (wk 3, 10, 18, and 26 postpartum). The method involved the application of the stable hydrogen isotope deuterium ((2)H) to the lactating dam. Drinking water intake and TWT decreased significantly with lactation stage, whether estimated by the isotope dilution technique or calculated from drinking water and water ingested from feeds. In contrast, lactation stage had no effect on dry matter intake, metabolizable energy (ME) intake, or the milk water fraction (i.e., the ratio between milk water excreted and TWT). The ratios between TWT measured and TWT estimated (by isotope dilution) did not differ with lactation stage and were close to 100% in all measurement weeks, indicating that the D(2)O dilution technique estimated TWT with high accuracy and only small variations. Calculating the required ME intakes for lactation from milk output data and gross energy content of milk revealed that, with increasing lactation stage, ME requirements per day for lactation decreased but remained constant per kilogram of milk output. Total measured ME intakes at different stages of lactation were similar to calculated ME intakes from published recommendation models for llamas. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A Multilaboratory, Multicountry Study To Determine Bedaquiline MIC Quality Control Ranges for Phenotypic Drug Susceptibility Testing

PubMed Central

Cirillo, Daniela M.; Hoffner, Sven; Ismail, Nazir A.; Kaur, Devinder; Lounis, Nacer; Metchock, Beverly; Pfyffer, Gaby E.; Venter, Amour

2016-01-01

The aim of this study was to establish standardized drug susceptibility testing (DST) methodologies and reference MIC quality control (QC) ranges for bedaquiline, a diarylquinoline antimycobacterial, used in the treatment of adults with multidrug-resistant tuberculosis. Two tier-2 QC reproducibility studies of bedaquiline DST were conducted in eight laboratories using Clinical Laboratory and Standards Institute (CLSI) guidelines. Agar dilution and broth microdilution methods were evaluated. Mycobacterium tuberculosis H37Rv was used as the QC reference strain. Bedaquiline MIC frequency, mode, and geometric mean were calculated. When resulting data occurred outside predefined CLSI criteria, the entire laboratory data set was excluded. For the agar dilution MIC, a 4-dilution QC range (0.015 to 0.12 μg/ml) centered around the geometric mean included 95.8% (7H10 agar dilution; 204/213 observations with one data set excluded) or 95.9% (7H11 agar dilution; 232/242) of bedaquiline MICs. For the 7H9 broth microdilution MIC, a 3-dilution QC range (0.015 to 0.06 μg/ml) centered around the mode included 98.1% (207/211, with one data set excluded) of bedaquiline MICs. Microbiological equivalence was demonstrated for bedaquiline MICs determined using 7H10 agar and 7H11 agar but not for bedaquiline MICs determined using 7H9 broth and 7H10 agar or 7H9 broth and 7H11 agar. Bedaquiline DST methodologies and MIC QC ranges against the H37Rv M. tuberculosis reference strain have been established: 0.015 to 0.12 μg/ml for the 7H10 and 7H11 agar dilution MICs and 0.015 to 0.06 μg/ml for the 7H9 broth microdilution MIC. These methodologies and QC ranges will be submitted to CLSI and EUCAST to inform future research and provide guidance for routine clinical bedaquiline DST in laboratories worldwide. PMID:27654337

On the various forms of the energy equation for a dilute, monatomic mixture of nonreacting gases

NASA Technical Reports Server (NTRS)

Kennedy, Christopher A.

1994-01-01

In the case of gas mixtures, the governing equations become rather formidable and a complete listing of the equations in their various forms and methods to evaluate the transport coefficients is difficult to find. This paper seeks to compile common, as well as less well known, results in a single document. Various relationships between equations describing conservation of energy for a dilute, monatomic, nonreacting gas in local equilibrium are provided. The gas is treated as nonrelativistic, not subject to magnetic or electric fields, or radiative effects.

Hydrocarbon synthesis catalyst and method of preparation

DOEpatents

Sapienza, R.S.; Sansone, M.J.; Slegeir, W.A.R.

1983-08-02

A catalyst for the synthesis of hydrocarbons from carbon monoxide and hydrogen composed of palladium or platinum and cobalt supported on a solid phase is disclosed. The catalyst is prepared by heating a heterogeneous component of the palladium or platinum deposited on the solid support in a solution of cobalt carbonyl or precursors thereof. The catalyst exhibits excellent activity, stability in air, and produces highly desirable product fractions even with dilute gaseous reactants. The catalyst is preferably used in dilute slurry form, which is desirable from a heat transfer standpoint. 9 figs.

Hydrocarbon synthesis catalyst and method of preparation

DOEpatents

Sapienza, Richard S.; Sansone, Michael J.; Slegeir, William A. R.

1983-08-02

A catalyst for the synthesis of hydrocarbons from carbon monoxide and hydrogen composed of palladium or platinum and cobalt supported on a solid phase is disclosed. The catalyst is prepared by heating a heterogeneous component of the palladium or platinum deposited on the solid support in a solution of cobalt carbonyl or precursors thereof. The catalyst exhibits excellent activity, stability in air, and produces highly desirable product fractions even with dilute gaseous reactants. The catalyst is preferably used in dilute slurry form, which is desirable from a heat transfer standpoint.

Detection of antimicrobial activity of banana peel (Musa paradisiaca L.) on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study.

PubMed

Kapadia, Suraj Premal; Pudakalkatti, Pushpa S; Shivanaikar, Sachin

2015-01-01

Banana is used widely because of its nutritional values. In past, there are studies that show banana plant parts, and their fruits can be used to treat the human diseases. Banana peel is a part of banana fruit that also has the antibacterial activity against microorganisms but has not been studied extensively. Since, there are no studies that relate the antibacterial activity of banana peel against periodontal pathogens. Hence, the aim of this study is to determine the antimicrobial activity of banana peel extract on Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). Standard strains of P. gingivalis and A. actinomycetemcomitans were used in this study which was obtained from the in-house bacterial bank of Department of Molecular Biology and Immunology at Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre. The banana peel extract was prepared, and the antibacterial activity was assessed using well agar diffusion method and minimum inhibitory concentration was assessed using serial broth dilution method. In the current study, both the tested microorganisms showed antibacterial activity. In well diffusion method, P. gingivalis and A. actinomycetemcomitans showed 15 mm and 12 mm inhibition zone against an alcoholic extract of banana peel, respectively. In serial broth dilution method P. gingivalis and A. actinomycetemcomitans were sensitive until 31.25 μg/ml dilutions. From results of the study, it is suggested that an alcoholic extract of banana peel has antimicrobial activity against P. gingivalis and A. actinomycetemcomitans.

Detection of antimicrobial activity of banana peel (Musa paradisiaca L.) on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study

PubMed Central

Kapadia, Suraj Premal; Pudakalkatti, Pushpa S.; Shivanaikar, Sachin

2015-01-01

Introduction and Aim: Banana is used widely because of its nutritional values. In past, there are studies that show banana plant parts, and their fruits can be used to treat the human diseases. Banana peel is a part of banana fruit that also has the antibacterial activity against microorganisms but has not been studied extensively. Since, there are no studies that relate the antibacterial activity of banana peel against periodontal pathogens. Hence, the aim of this study is to determine the antimicrobial activity of banana peel extract on Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). Material and Methods: Standard strains of P. gingivalis and A. actinomycetemcomitans were used in this study which was obtained from the in-house bacterial bank of Department of Molecular Biology and Immunology at Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Sciences and Research Centre. The banana peel extract was prepared, and the antibacterial activity was assessed using well agar diffusion method and minimum inhibitory concentration was assessed using serial broth dilution method. Results: In the current study, both the tested microorganisms showed antibacterial activity. In well diffusion method, P. gingivalis and A. actinomycetemcomitans showed 15 mm and 12 mm inhibition zone against an alcoholic extract of banana peel, respectively. In serial broth dilution method P. gingivalis and A. actinomycetemcomitans were sensitive until 31.25 μg/ml dilutions. Conclusion: From results of the study, it is suggested that an alcoholic extract of banana peel has antimicrobial activity against P. gingivalis and A. actinomycetemcomitans. PMID:26681854

Dilution Confusion: Conventions for Defining a Dilution

ERIC Educational Resources Information Center

Fishel, Laurence A.

2010-01-01

Two conventions for preparing dilutions are used in clinical laboratories. The first convention defines an "a:b" dilution as "a" volumes of solution A plus "b" volumes of solution B. The second convention defines an "a:b" dilution as "a" volumes of solution A diluted into a final volume of "b". Use of the incorrect dilution convention could affect…

RADIOISOTOPES USED IN PHARMACY. 5. IONIZING RADIATION IN PHARMACEUTICAL ANALYSIS (in Danish)

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

Kristensen, K.

1962-09-01

The use of radioisotope methods for analyzing drugs is reviewed. It is pointed out that heretofore most methods have been based on isotope dilution principles whereas in the future radioactivation analysis, especially with neutron sources, offers great possibilities. (BBB)

21 CFR 177.1570 - Poly-1-butene resins and butene/ethylene copolymers.

Code of Federal Regulations, 2014 CFR

2014-04-01

...) Viscosity. Poly-1-butene resins and the butene/ethylene copolymers have an intrinsic viscosity 1.0 to 3.2 as determined by ASTM method D1601-78, “Standard Test Method for Dilute Solution Viscosity of Ethylene Polymers...

FACTORS INFLUENCING THE ABILITY OF ISOLATED CELL NUCLEI TO FORM GELS IN DILUTE ALKALI

PubMed Central

Dounce, Alexander L.; Monty, Kenneth J.

1955-01-01

1. Known methods for isolating cell nuclei are divided into two classes, depending on whether or not the nuclei are capable of forming gels in dilute alkali or strong saline solutions. Methods which produce nuclei that can form gels apparently prevent the action of an intramitochondrial enzyme capable of destroying the gel-forming capacity of the nuclei. Methods in the other class are believed to permit this enzyme to act on the nuclei during the isolation procedure, causing detachment of DNA from some nuclear constituent (probably protein). 2. It is shown that heating in alkaline solution and x-irradiation can destroy nuclear gels. Heating in acid or neutral solutions can destroy the capacity of isolated nuclei to form gels. 3. Chemical and biological evidence is summarized in favor of the hypothesis that DNA is normally bound firmly to some nuclear component by non-ionic linkages. PMID:14381437

Electron-nuclear spin dynamics of Ga centers in GaAsN dilute nitride semiconductors probed by pump-probe spectroscopy

NASA Astrophysics Data System (ADS)

Sandoval-Santana, J. C.; Ibarra-Sierra, V. G.; Azaizia, S.; Carrère, H.; Bakaleinikov, L. A.; Kalevich, V. K.; Ivchenko, E. L.; Marie, X.; Amand, T.; Balocchi, A.; Kunold, A.

2018-03-01

We propose an experimental procedure to track the evolution of electronic and nuclear spins in Ga2+ centers in GaAsN dilute semiconductors. The method is based on a pump-probe scheme that enables to monitor the time evolution of the three components of the electronic and nuclear spin variables. In contrast to other characterization methods, as nuclear magnetic resonance, this one only needs moderate magnetic fields (B≈ 10 mT), and does not require microwave irradiation. Specifically, we carry out a series of tests for different experimental conditions in order to optimize the procedure for maximum sensitivity in the measurement of the circular degree of polarization. Based on previous experimental results and the theoretical calculations presented here, we estimate that the method could yield a time resolution of about 10ps.

A competitive chemiluminescence enzyme immunoassay for rapid and sensitive determination of enrofloxacin

NASA Astrophysics Data System (ADS)

Yu, Fei; Wu, Yongjun; Yu, Songcheng; Zhang, Huili; Zhang, Hongquan; Qu, Lingbo; Harrington, Peter de B.

With alkaline phosphatase (ALP)-adamantane (AMPPD) system as the chemiluminescence (CL) detection system, a highly sensitive, specific and simple competitive chemiluminescence enzyme immunoassay (CLEIA) was developed for the measurement of enrofloxacin (ENR). The physicochemical parameters, such as the chemiluminescent assay mediums, the dilution buffer of ENR-McAb, the volume of dilution buffer, the monoclonal antibody concentration, the incubation time, and other relevant variables of the immunoassay have been optimized. Under the optimal conditions, the detection linear range of 350-1000 pg/mL and the detection limit of 0.24 ng/mL were provided by the proposed method. The relative standard deviations were less than 15% for both intra and inter-assay precision. This method has been successfully applied to determine ENR in spiked samples with the recovery of 103%-96%. It showed that CLEIA was a good potential method in the analysis of residues of veterinary drugs after treatment of related diseases.

Microfocused Ultrasound with Visualization and Calcium Hydroxylapatite for Improving Skin Laxity and Cellulite Appearance

PubMed Central

Pereira, Gregório

2017-01-01

Background: Age-associated skin laxity contributes to worsening of cellulite appearance. This study evaluated the effects of microfocused ultrasound with visualization (MFU-V; Ultherapy) in combination with diluted calcium hydroxylapatite (CaHA; Radiesse) on cellulite appearance and on neocollagenesis. Methods: Twenty women (18–55 years old) with skin laxity and moderate-to-severe cellulite on the buttocks and thighs were retrospectively enrolled. MFU-V was applied using 4 and 7 MHz transducers (25 lines/transducer/site) and immediately followed by subdermal CaHA injection (1 ml/buttock or thigh). Photographs at baseline and 90 days were assessed by 2 independent, blinded evaluators using a 5-item cellulite severity scale. One subject scheduled for thighplasty received treatment with 6 different CaHA dilutions (0.3 ml/5 cm2) followed by MFU-V. Tissue specimens from each dilution site were examined under polarized light microscopy to assess neocollagenesis. Results: Both evaluators reported statistically significant improvements compared with baseline for each item on the cellulite severity scale (P < 0.001) with a 4.5-point improvement in mean overall score (P < 0.001) after a single MFU-V/CaHA treatment. At 90 days, histologic analysis showed peak neocollagenesis in samples treated with the 1:1 dilution, whether with CaHA alone or in combination with MFU-V. The highest conversion of collagen type III into collagen type I at month 3 occurred in samples injected with 1:1 and 1:0.6 CaHA dilutions without subsequent MFU-V treatment. Both procedures were well tolerated, and subject satisfaction was high. Conclusions: Combination treatment with MFU-V and diluted CaHA is effective for improving skin laxity and the appearance of cellulite on the buttocks and upper thighs. PMID:28831339

The use of animal models in homeopathic research--a review of 2010-2014 PubMed indexed papers.

PubMed

Bonamin, Leoni Villano; Cardoso, Thayná Neves; de Carvalho, Aloísio Cunha; Amaral, Juliana Gimenez

2015-10-01

In the 1990s, a study was performed on the effects of highly diluted thyroxine on frog metamorphosis. This model represented one of the most discussed examples of the biological effects of high dilutions over the next two decades. In 2010, another critical conceptual review of the use of animal models in homeopathy and high-dilution research was published. The main contribution of these studies was the elucidation of the biological features and phenomenology of the effects of high dilutions on living systems, representing an important step forward in our understanding of the mechanisms of action of homeopathic medicines. We performed a further review of this line of investigation using the same methods. Fifty-three articles that were indexed in the PubMed database and used 12 different animal species were systematically evaluated. Only a fraction of the studies (29/53) reported herein were performed with "ultra high" dilutions. The other studies were performed with dilutions in ranges below 10(-23) (14/53 articles) or commercial complexes (10/53 articles). Only two articles reported negative results; both used in vivo protocols to test commercial complexes, one in fish and one in bees. The quality of the employed techniques improved in 2010-2014 compared with the studies that were reviewed previously in 2010, with the inclusion of more ethically refined protocols, including in vitro primary cell cultures and ex vivo studies (10/53 articles), often with three or more replicates and analyses of epigenetic mechanisms that were previously unknown in 2010. In our updated review of the past 5 years, we found further demonstrations of the biological effects of homeopathy using more refined animal models and in vitro techniques. Copyright © 2015 The Faculty of Homeopathy. Published by Elsevier Ltd. All rights reserved.

Ki-67 staining for determination of rhesus macaque T cell proliferative responses ex vivo1

PubMed Central

Shedlock, Devon J.; Talbott, Kendra T.; Morrow, Matthew P.; Ferraro, Bernadette; Hokey, David A.; Muthumani, Karuppiah; Weiner, David B.

2010-01-01

The capacity for robust proliferation upon re-infection is a hallmark of adaptive immunity and the basis of vaccination. A widely used animal model for the study of human disease is the rhesus macaque (RM), where capacity for proliferation can be assessed ex vivo using carboxyfluorescein succinimidyl ester (CFSE)-based dilution assays. However, we show over the course of the standard ex vivo proliferation assay that CFSE-labeling at commonly-used dye concentrations induces significant cell death, but that this phenomenon is dose-dependent. Here we describe an alternative, semi-quantitative method for estimating T cell proliferative responses that avoids the putative biases associated with chemical modification. RM peripheral blood mononuclear cells were stimulated ex vivo with cognate peptides for five days, immunostained for intracellular Ki-67, and then analyzed by flow cytometry. We describe a gating strategy using Ki-67 and side light scatter, also a marker of blastogenesis, which correlates strongly with data from CFSE dilution. We show that this method is a valid tool for measuring RM antigen-specific cellular proliferation ex vivo and can be used as an alternative to CFSE dilution assays. PMID:20104580

Investigation of Latent Traces Using Infrared Reflectance Hyperspectral Imaging

NASA Astrophysics Data System (ADS)

Schubert, Till; Wenzel, Susanne; Roscher, Ribana; Stachniss, Cyrill

2016-06-01

The detection of traces is a main task of forensics. Hyperspectral imaging is a potential method from which we expect to capture more fluorescence effects than with common forensic light sources. This paper shows that the use of hyperspectral imaging is suited for the analysis of latent traces and extends the classical concept to the conservation of the crime scene for retrospective laboratory analysis. We examine specimen of blood, semen and saliva traces in several dilution steps, prepared on cardboard substrate. As our key result we successfully make latent traces visible up to dilution factor of 1:8000. We can attribute most of the detectability to interference of electromagnetic light with the water content of the traces in the shortwave infrared region of the spectrum. In a classification task we use several dimensionality reduction methods (PCA and LDA) in combination with a Maximum Likelihood classifier, assuming normally distributed data. Further, we use Random Forest as a competitive approach. The classifiers retrieve the exact positions of labelled trace preparation up to highest dilution and determine posterior probabilities. By modelling the classification task with a Markov Random Field we are able to integrate prior information about the spatial relation of neighboured pixel labels.

Detection and isolation of rare cells by 2-step enrichment high-speed flow cytometry/cell sorting and single cell LEAP laser ablation

NASA Astrophysics Data System (ADS)

Zordan, M. D.; Leary, James F.

2011-02-01

The clonal isolation of rare cells, especially cancer and stem cells, in a population is important to the development of improved medical treatment. We have demonstrated that the Laser-Enabled Analysis and Processing (LEAP, Cyntellect Inc., San Diego, CA) instrument can be used to efficiently produce single cell clones by photoablative dilution. Additionally, we have also shown that cells present at low frequencies can be cloned by photoablative dilution after they are pre-enriched by flow cytometry based cell sorting. Circulating tumor cells were modeled by spiking isolated peripheral blood cells with cells from the lung carcinoma cell line A549. Flow cytometry based cell sorting was used to perform an enrichment sort of A549 cells directly into a 384 well plate. Photoablative dilution was performed with the LEAPTM instrument to remove any contaminating cells, and clonally isolate 1 side population cell per well. We were able to isolate and grow single clones of side population cells using this method at greater than 90% efficiency. We have developed a 2 step method that is able to perform the clonal isolation of rare cells based on a medically relevant functional phenotype.

Application of a data reconciliation method to the stoichiometric analysis of Fibrobacter succinogenes growth.

PubMed

Guiavarch, Erell; Pons, Agnes; Creuly, Catherine; Dussap, Claude-Gilles

2008-12-01

Fibrobacter succinogenes S85, a strictly anaerobic Gram-negative bacterium, was grown in continuous culture in a bioreactor at different dilution rates (0.02 to 0.092 h(-1)) on a fully synthetic culture medium with glucose as carbon source. Glucose and ammonium sulfate consumption, as well as biomass, succinate, acetate, formate, and carbohydrate production were regularly measured. The relevant biomass elemental compositions were established for each dilution rate. Robustness of the experimental information was checked by C and N mass balances estimation, which were satisfactory. A detailed overall stoichiometry analysis of the process, including all substrates and products of the culture, was proposed. Online and off-line parameters measured during the culture brought a large number of data which were weighted by their respective variance associated to the measured value. The material balance resulted in an overdetermined linear system of equations made of weighted relationships including experimental data, elemental balances (C, H, O, N, S, Na), and an additional constraint. The mass balances involved in stoichiometric equations were solved using data reconciliation and linear algebra methods to take into account error measurements. This methodology allowed to establish the overall stoichiometric equation for each dilution rate studied.

Body Fat and Muscle Mass as Functions of Body Water

ERIC Educational Resources Information Center

Sutton, R. A.; Miller, Carolyn

2007-01-01

Hydrostatic weighing and chemical dilution are well accepted methods for measuring body composition. Recently, Dual Energy X-ray Absorptiometry (DEXA) has become the preferred method. The two compartment algorithms used by these methods assume a fixed constant for lean body tissue. This constant has long been suspect of variations due to many…

Comparison of Analytical Methods for the Determination of Uranium in Seawater Using Inductively Coupled Plasma Mass Spectrometry

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

Wood, Jordana R.; Gill, Gary A.; Kuo, Li-Jung

2016-04-20

Trace element determinations in seawater by inductively coupled plasma mass spectrometry are analytically challenging due to the typically very low concentrations of the trace elements and the potential interference of the salt matrix. In this study, we did a comparison for uranium analysis using inductively coupled plasma mass spectrometry (ICP-MS) of Sequim Bay seawater samples and three seawater certified reference materials (SLEW-3, CASS-5 and NASS-6) using seven different analytical approaches. The methods evaluated include: direct analysis, Fe/Pd reductive precipitation, standard addition calibration, online automated dilution using an external calibration with and without matrix matching, and online automated pre-concentration. The methodmore » which produced the most accurate results was the method of standard addition calibration, recovering uranium from a Sequim Bay seawater sample at 101 ± 1.2%. The on-line preconcentration method and the automated dilution with matrix-matched calibration method also performed well. The two least effective methods were the direct analysis and the Fe/Pd reductive precipitation using sodium borohydride« less

Lipid and Creatinine Adjustment to Evaluate Health Effects of Environmental Exposures.

PubMed

O'Brien, Katie M; Upson, Kristen; Buckley, Jessie P

2017-03-01

Urine- and serum-based biomarkers are useful for assessing individuals' exposure to environmental factors. However, variations in urinary creatinine (a measure of dilution) or serum lipid levels, if not adequately corrected for, can directly impact biomarker concentrations and bias exposure-disease association measures. Recent methodological literature has considered the complex relationships between creatinine or serum lipid levels, exposure biomarkers, outcomes, and other potentially relevant factors using directed acyclic graphs and simulation studies. The optimal measures of urinary dilution and serum lipids have also been investigated. Existing evidence supports the use of covariate-adjusted standardization plus creatinine adjustment for urinary biomarkers and standardization plus serum lipid adjustment for lipophilic, serum-based biomarkers. It is unclear which urinary dilution measure is best, but all serum lipid measures performed similarly. Future research should assess methods for pooled biomarkers and for studying diseases and exposures that affect creatinine or serum lipids directly.

Identification and Quantitation of Potent Odorants in Spearmint Oils.

PubMed

Kelley, Lauren E; Cadwallader, Keith R

2018-03-14

Potent odorants in Native spearmint, Scotch spearmint, and Macho mint oils were determined by the combined use of gas chromatography-olfactometry (GCO), gas chromatography-mass spectrometry (GC-MS), and aroma extract dilution analysis (AEDA). Of the 85 odorants detected, ( R)-(-)-carvone was the most potent odorant in all three spearmint oils. Additional predominant odorants in all spearmint oils included eugenol, ethyl ( S)-(+)-2-methylbutanoate, ( E)-β-damascenone, and (3 E,5 Z)-1,3,5-undecatriene. Forty-six compounds were quantitated using various methods, including 19 by gas chromatography with flame ionization detection (GC-FID), 20 by stable isotope dilution analysis (SIDA), and 14 by GCO dilution analysis. Concentrations were used to calculate the odor activity values (OAVs) for predominant odorants in the oils. Among the compounds quantitated, those with the highest OAVs were ( R)-(-)-carvone, 1,8-cineole, ( E, Z)-2,6-nonadienal, ( E)-β-damascenone, and (3 E,5 Z)-1,3,5-undecatriene.

Lipid and Creatinine Adjustment to Evaluate Health Effects of Environmental Exposures

PubMed Central

O’Brien, Katie M.; Upson, Kristen; Buckley, Jessie P.

2017-01-01

Purpose of review Urine- and serum-based biomarkers are useful for assessing individuals’ exposure to environmental factors. However, variations in urinary creatinine (a measure of dilution) or serum lipid levels, if not adequately corrected for, can directly impact biomarker concentrations and bias exposure-disease association measures. Recent findings Recent methodological literature has considered the complex relationships between creatinine or serum lipid levels, exposure biomarkers, outcomes, and other potentially relevant factors using directed acyclic graphs and simulation studies. The optimal measures of urinary dilution and serum lipids have also been investigated. Summary Existing evidence supports the use of covariate-adjusted standardization plus creatinine adjustment for urinary biomarkers and standardization plus serum lipid adjustment for lipophilic, serum-based biomarkers. It is unclear which urinary dilution measure is best, but all serum lipid measures performed similarly. Future research should assess methods for pooled biomarkers and for studying diseases and exposures that affect creatinine or serum lipids directly. PMID:28097619

Spectroscopic studies of GTA welding plasmas. Temperature calculation and dilution measurement

NASA Astrophysics Data System (ADS)

Lacroix, D.; Boudot, C.; Jeandel, G.

1999-10-01

A spectroscopic study of the GTAW plasma-plume created during the welding of stainless steel and other materials (iron, nickel and chromium) has been carried out. The spectra of these plasmas have been studied for several welding parameters. Temperature calculations are based on the observation of relative intensities and shapes of the emission peaks. We assume that the plasma is in local thermal equilibrium. The temperature is calculated with the Boltzmann plot method from twelve iron emission lines (in the range 368 385 nm): it varies between 9650 and 12 100 K. Dilution experiments have been carried out. We checked the mixing of metals: during welding of two different metallic plates and during welding with an Inconel wire. Dilution is monitored following the intensity of some characteristic emission lines (chromium and nickel). Comparison of spectroscopic results and metallographic ones is made.

Effect of Low and Very Low Doses of Simple Phenolics on Plant Peroxidase Activity

PubMed Central

Malarczyk, Elżbieta; Kochmańska-Rdest, Janina; Paździoch-Czochra, Marzanna

2004-01-01

Changes in the activity of horseradish peroxidase resulting from an addition of ethanol water dilutions of 19 phenolic compounds were observed. For each compound, the enzyme activity was plotted against the degree of dilution expressed as n = –log100 (mol/L) in the range 0 ≤ n ≥ 20. All the curves showed sinusoidal activity, more or less regular, with two to four peaks on average. Each analyzed compound had a characteristic sinusoidal shape, which was constant for samples of peroxidase from various commercial firms. This was clearly visible after function fitting to experimental results based on the Marquadt–Levenberg algorithm using the least-squares method. Among the 19 phenolics, the highest amplitudes were observed for phenol and iso- and vanillate acids and aldehydes. The specific character of each of the analyzed curves offers a possibility of choosing proper dilutions of phenolic compound for activating or inhibiting of peroxidase activity. PMID:19330128

The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques.

PubMed

Sabattini, E; Bisgaard, K; Ascani, S; Poggi, S; Piccioli, M; Ceccarelli, C; Pieri, F; Fraternali-Orcioni, G; Pileri, S A

1998-07-01

To assess a newly developed immunohistochemical detection system, the EnVision++. A large series of differently processed normal and pathological samples and 53 relevant monoclonal antibodies were chosen. A chessboard titration assay was used to compare the results provided by the EnVision++ system with those of the APAAP, CSA, LSAB, SABC, and ChemMate methods, when applied either manually or in a TechMate 500 immunostainer. With the vast majority of the antibodies, EnVision++ allowed two- to fivefold higher dilutions than the APAAP, LSAB, SABC, and ChemMate techniques, the staining intensity and percentage of expected positive cells being the same. With some critical antibodies (such as the anti-CD5), it turned out to be superior in that it achieved consistently reproducible results with differently fixed or overfixed samples. Only the CSA method, which includes tyramide based enhancement, allowed the same dilutions as the EnVision++ system, and in one instance (with the anti-cyclin D1 antibody) represented the gold standard. The EnVision++ is an easy to use system, which avoids the possibility of disturbing endogenous biotin and lowers the cost per test by increasing the dilutions of the primary antibodies. Being a two step procedure, it reduces both the assay time and the workload.

Laminar Flame Velocity and Temperature Exponent of Diluted DME-Air Mixture

NASA Astrophysics Data System (ADS)

Naseer Mohammed, Abdul; Anwar, Muzammil; Juhany, Khalid A.; Mohammad, Akram

2017-03-01

In this paper, the laminar flame velocity and temperature exponent diluted dimethyl ether (DME) air mixtures are reported. Laminar premixed mixture of DME-air with volumetric dilutions of carbon dioxides (CO2) and nitrogen (N2) are considered. Experiments were conducted using a preheated mesoscale high aspect-ratio diverging channel with inlet dimensions of 25 mm × 2 mm. In this method, flame velocities are extracted from planar flames that were stabilized near adiabatic conditions inside the channel. The flame velocities are then plotted against the ratio of mixture temperature and the initial reference temperature. A non-linear power law regression is observed suitable. This regression analysis gives the laminar flame velocity at the initial reference temperature and temperature exponent. Decrease in the laminar flame velocity and increase in temperature exponent is observed for CO2 and N2 diluted mixtures. The addition of CO2 has profound influence when compared to N2 addition on both flame velocity and temperature exponent. Numerical prediction of the similar mixture using a detailed reaction mechanism is obtained. The computational mechanism predicts higher magnitudes for laminar flame velocity and smaller magnitudes of temperature exponent compared to experimental data.

Infiltration as Ventilation: Weather-Induced Dilution

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

Sherman, Max H.; Turner, William J.N.; Walker, Iain S.

The purpose of outdoor air ventilation is to dilute or remove indoor contaminants to which occupants are exposed. It can be provided by mechanical or natural means. In most homes, especially older homes, weather-driven infiltration provides the dominant fraction of the total ventilation. As we seek to provide good indoor air quality at minimum energy cost, it is important to neither over-ventilate nor under-ventilate. Thus, it becomes critically important to evaluate correctly the contribution infiltration makes to the total outdoor air ventilation rate. Because weather-driven infiltration is dependent on building air leakage and weather-induced pressure differences, a given amount ofmore » air leakage will provide different amounts of infiltration. Varying rates of infiltration will provide different levels of contaminant dilution and hence effective ventilation. This paper derives these interactions and then calculates the impact of weather-driven infiltration for different climates. A new “N-factor” is introduced to provide a convenient method for calculating the ventilation contribution of infiltration for over 1,000 locations across North America. The results of this work could be used in indoor air quality standards (specifically ASHRAE 62.2) to account for the contribution of weather-driven infiltration towards the dilution of indoor pollutants.« less

Multi-channel Auto-dilution System for Remote Continuous Monitoring of High Soil-CO2 Fluxes

NASA Astrophysics Data System (ADS)

Barr, J. L.; Amonette, J. E.

2008-12-01

We describe a novel field instrument that takes input from up to 27 soil flux chambers and measures flux using the steady-state method. CO2 concentrations are determined with an infrared gas analyzer (IRGA, 0- 3000 ppmv range) with corrections for temperature, barometric pressure, and moisture content. The concentrations are monitored during data collection and, if they exceed the range of the IRGA, a stepped dilution program is automatically implemented that allows up to 50-fold dilution of the incoming gas stream with N2 supplied by boil-off from a large dewar. The upper concentration limit of the system with dilution is extended to at least 150,000 ppmv CO2. The data are stored on a datalogger having a cellular modem connection that allows remote control of the system as well as transmittal of data. The system is designed to operate for six weeks with no on-site maintenance required. Longer periods are possible with modifications to allow on-site generation of N2 from air. Example data from a recent CO2 test injection at the Zero- Emission Research and Technology (ZERT) field site in Bozeman, MT are presented.

Translating Thermal Response of Triblock Copolymer Assemblies in Dilute Solution to Macroscopic Gelation and Phase Separation

DOE PAGES

Sun, Zhe; Tian, Ye; Hom, Wendy L.; ...

2016-12-28

The thermal response of semi-dilute solutions (5 w/w%) of two amphiphilic thermoresponsive poly(ethylene oxide)-b-poly(N,N-diethylacrylamide)-b-poly(N,N-dibutylacrylamide) (PEO 45-PDEAm x-PDBAm 12) triblock copolymers, which differ only in the size of the central responsive block, in water was examined in this paper. Aqueous PEO45-PDEAm41-PDBAm12 solutions, which undergo a thermally induced sphere-to-worm transition in dilute solution, were found to reversibly form soft (G'≈10 Pa) free-standing physical gels after 10 min at 55 °C. PEO 45-PDEAm 89-PDBAm 12 copolymer solutions, which undergo a thermally induced transition from spheres to large compound micelles (LCM) in dilute solution, underwent phase separation after heating at 55 °C for 10more » min owing to sedimentation of LCMs. The reversibility of LCM formation was investigated as a non-specific method for removal of a water-soluble dye from aqueous solution. Finally, the composition and size of the central responsive block in these polymers dictate the microscopic and macroscopic response of the polymer solutions as well as the rates of transition between assemblies.« less

Platelet counting using the Coulter electronic counter.

PubMed

Eggleton, M J; Sharp, A A

1963-03-01

A method for counting platelets in dilutions of platelet-rich plasm using the Coulter electronic counter is described.(1) The results obtained show that such platelet counts are at least as accurate as the best methods of visual counting. The various technical difficulties encountered are discussed.

Ultrasonication extraction and gel permeation chromatography clean-up for the determination of polycyclic aromatic hydrocarbons in edible oil by an isotope dilution gas chromatography–mass spectrometry.

PubMed

Wang, Jian-Hua; Guo, Cui

2010-07-09

An analytical method for the determination of US EPA priority pollutant 16 polycyclic aromatic hydrocarbons (PAHs) in edible oil was developed by an isotope dilution gas chromatography-mass spectrometry (GC-MS). Extraction was performed with ultrasonication mode using acetonitrile as solvent, and subsequent clean-up was applied using narrow gel permeation chromatographic column. Three deuterated PAHs surrogate standards were used as internal standards for quantification and analytical quality control. The limits of quantification (LOQs) were globally below 0.5 ng/g, the recoveries were in the range of 81-96%, and the relative standard deviations (RSDs) were lower than 20%. Further trueness assessment of the method was also verified through participation in international cocoa butter proficiency test (T0638) organised by the FAPAS with excellent results in 2008. The results obtained with the described method were satisfying (z ≤ 2). The method has been applied to determine PAH in real edible oil samples.

Cloning of Plasmodium falciparum by single-cell sorting.

PubMed

Miao, Jun; Li, Xiaolian; Cui, Liwang

2010-10-01

Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two-dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples. Copyright 2010 Elsevier Inc. All rights reserved.

A new self-shielding method based on a detailed cross-section representation in the resolved energy domain

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

Saygin, H.; Hebert, A.

The calculation of a dilution cross section {bar {sigma}}{sub e} is the most important step in the self-shielding formalism based on the equivalence principle. If a dilution cross section that accurately characterizes the physical situation can be calculated, it can then be used for calculating the effective resonance integrals and obtaining accurate self-shielded cross sections. A new technique for the calculation of equivalent cross sections based on the formalism of Riemann integration in the resolved energy domain is proposed. This new method is compared to the generalized Stamm`ler method, which is also based on an equivalence principle, for a two-regionmore » cylindrical cell and for a small pressurized water reactor assembly in two dimensions. The accuracy of each computing approach is obtained using reference results obtained from a fine-group slowing-down code named CESCOL. It is shown that the proposed method leads to slightly better performance than the generalized Stamm`ler approach.« less

Simultaneous determination of the quantity and isotopic ratios of uranium in individual micro-particles by isotope dilution thermal ionization mass spectrometry (ID-TIMS).

PubMed

Park, Jong-Ho; Choi, Eun-Ju

2016-11-01

A method to determine the quantity and isotopic ratios of uranium in individual micro-particles simultaneously by isotope dilution thermal ionization mass spectrometry (ID-TIMS) has been developed. This method consists of sequential sample and spike loading, ID-TIMS for isotopic measurement, and application of a series of mathematical procedures to remove the contribution of uranium in the spike. The homogeneity of evaporation and ionization of uranium content was confirmed by the consistent ratio of n((233)U)/n((238)U) determined by TIMS measurements. Verification of the method was performed using U030 solution droplets and U030 particles. Good agreements of resulting uranium quantity, n((235)U)/n((238)U), and n((236)U)/n((238)U) with the estimated or certified values showed the validity of this newly developed method for particle analysis when simultaneous determination of the quantity and isotopic ratios of uranium is required. Copyright © 2016 Elsevier B.V. All rights reserved.

Measurement of biochemical oxygen demand of the leachates.

PubMed

Fulazzaky, Mohamad Ali

2013-06-01

Biochemical oxygen demand (BOD) of the leachates originally from the different types of landfill sites was studied based on the data measured using the two manometric methods. The measurements of BOD using the dilution method were carried out to assess the typical physicochemical and biological characteristics of the leachates together with some other parameters. The linear regression analysis was used to predict rate constants for biochemical reactions and ultimate BOD values of the different leachates. The rate of a biochemical reaction implicated in microbial biodegradation of pollutants depends on the leachate characteristics, mass of contaminant in the leachate, and nature of the leachate. Character of leachate samples for BOD analysis of using the different methods may differ significantly during the experimental period, resulting in different BOD values. This work intends to verify effect of the different dilutions for the manometric method tests on the BOD concentrations of the leachate samples to contribute to the assessment of reaction rate and microbial consumption of oxygen.

A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores.

PubMed

Nerandzic, Michelle M; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J

2016-01-01

Background.  Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5-2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods.  We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results.  Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200-2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions.  Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin.

Cow's Milk Desensitization in Anaphylactic Patients: A New Personalized-dose Method.

PubMed

Babaie, Delara; Nabavi, Mohammad; Arshi, Saba; Mesdaghi, Mehrnaz; Chavoshzadeh, Zahra; Bemanian, Mohammad Hasan; Tafakori, Mitra; Amirmoini, Mehrdad; Esmailzadeh, Hosein; Molatefi, Rasoul; Rekabi, Mahsa; Akbarpour, Nadieh; Masoumi, Farimah; Fallahpour, Morteza

2017-02-01

Cow's milk allergy (CMA) is the most frequent food allergy in children and oral immunotherapy (OIT) is a promising approach for treatment of patients. The most challenging cases are anaphylactic with coexisting asthma and proposing safe protocols is crucial especially in high risk groups. Considering that CMA varies among patients, an individualized OIT protocol would be beneficial to achieve a safer and more efficient method of desensitization. 18 children more than 3 years of age with IgE-mediated CMA were enrolled. CMA was confirmed by positive skin prick test (SPT) and positive oral food challenge (OFC) and 60% of individuals had a convincing history of persistent asthma. SPT with milk extracts, whole fresh milk and serially diluted milk concentrations were performed.  The dilution of milk that induced 3-5 mm of wheal in each individual was selected as the starting dilution for OIT. Desensitization began by 1 drop of the defined dilution and continued increasingly. Overall, 16 out of 18 children (88.8%) achieved the daily intake of 120 mL of milk. Four out of these 16 children accomplished the protocol without any adverse allergic reactions. 12 patients experienced mild to severe reactions. Wheal and erythema in SPT (p≤0.001), and sIgE (p≤0.003) to most milk allergens were significantly decreased following desensitization. We successfully desensitized 16 of 18 children with IgE-mediated CMA by individualized desensitization protocol. Individualizing the OIT protocol would be helpful to save time and perhaps to relieve the allergic symptoms after ingesting cow's milk intake.

Kinetic theory of weakly ionized dilute gas of hydrogen-like atoms of the first principles of quantum statistics and dispersion laws of eigenwaves

NASA Astrophysics Data System (ADS)

Slyusarenko, Yurii V.; Sliusarenko, Oleksii Yu.

2017-11-01

We develop a microscopic approach to the construction of the kinetic theory of dilute weakly ionized gas of hydrogen-like atoms. The approach is based on the statements of the second quantization method in the presence of bound states of particles. The basis of the derivation of kinetic equations is the method of reduced description of relaxation processes. Within the framework of the proposed approach, a system of common kinetic equations for the Wigner distribution functions of free oppositely charged fermions of two kinds (electrons and cores) and their bound states—hydrogen-like atoms— is obtained. Kinetic equations are used to study the spectra of elementary excitations in the system when all its components are non-degenerate. It is shown that in such a system, in addition to the typical plasma waves, there are longitudinal waves of matter polarization and the transverse ones with a behavior characteristic of plasmon polaritons. The expressions for the dependence of the frequencies and Landau damping coefficients on the wave vector for all branches of the oscillations discovered are obtained. Numerical evaluation of the elementary perturbation parameters in the system on an example of a weakly ionized dilute gas of the 23Na atoms using the D2-line characteristics of the natrium atom is given. We note the possibility of using the results of the developed theory to describe the properties of a Bose condensate of photons in the diluted weakly ionized gas of hydrogen-like atoms.

Red blood cell populations and membrane levels of peroxiredoxin 2 as candidate biomarkers to reveal blood doping

PubMed Central

Marrocco, Cristina; Pallotta, Valeria; D’Alessandro, Angelo; Alves, Gilda; Zolla, Lello

2012-01-01

Background Blood doping represents one main trend in doping strategies. Blood doping refers to the practice of boosting the number of red blood cells (RBCs) in the bloodstream in order to enhance athletic performance, by means of blood transfusions, administration of erythropoiesis-stimulating substances, blood substitutes, natural or artificial altitude facilities, and innovative gene therapies. While detection of recombinant EPO and homologous transfusion is already feasible through electrophoretic, mass spectrometry or flow cytometry-based approaches, no method is currently available to tackle doping strategies relying on autologous transfusions. Materials and methods. We exploited an in vitro model of autologous transfusion through a 1:10 dilution of concentrated RBCs after 30 days of storage upon appropriate dilution in freshly withdrawn RBCs from the same donor. Western blot towards membrane Prdx2 and Percoll density gradients were exploited to assess their suitability as biomarkers of transfusion. Results Membrane Prdx2 was visible in day 30 samples albeit not in day 0, while it was still visible in the 1:10 dilution of day 30 in day 0 RBCs. Cell gradients also highlighted changes in the profile of the RBC subpopulations upon dilution of stored RBCs in the fresh ones. Discussion. From this preliminary in vitro investigation it emerges that Prdx2 and RBC populations might be further tested as candidate biomarkers of blood doping through autologous transfusion, though it is yet to be assessed whether the kinetics in vivo of Prdx2 exposure in the membrane of transfused RBCs will endow a sufficient time-window to allow reliable anti-doping testing. PMID:22890272

Identification of hydrolyzed inulin syrup and high-fructose corn syrup in apple juice by capillary gas chromatography: PVM 4:1999.

PubMed

Low, N H; McLaughlin, M A; Page, S W; Canas, B J; Brause, A R; Low, N H

2001-01-01

A peer-verified, gas chromatographic (GC) method is presented for the identification of hydrolyzed inulin syrup (HIS) and high-fructose corn syrup (HFCS) in apple juice. The procedure involves determining the Brix value of the apple juice or apple juice concentrate and preparing a dilution of the test sample to 5.5 degrees Brix. A 100 microL aliquot of the 5.5 degrees Brix test solution is then freeze-dried in a GC autosampler vial. The sugars in the freeze-dried residue are converted to trimethylsilyl derivatives, by the addition of an appropriate silylation reagent, and the vial is heated at 75 degrees C for 30 min. After derivatization, the solution is introduced into a gas chromatograph where the analytes are separated on a 30 m, 0.25 mm id DB-5 column. The method can use hydrogen, helium, or nitrogen as the carrier gas. The analytes and marker compounds are measured by use of a flame ionization detecone of the 2 syrups at 2 levels. Dilution was ascertained by the presence of retrograde sugar markers found in the 2 sugar syrups. All 3 laboratories involved in the study were able to identify the correct diluent in the blind, randomly coded, apple juice test portions. The levels of dilution in the test portions were 0, 6.9% (HIS), 16.0% (HIS), 8.1% (HFCS), and 17.0% (HFCS). No false positive results were reported. Quantitative conclusions can be drawn when the same syrup is used for dilution and as a reference standard.

Acute and early life stage toxicity of industrial effluent on Japanese medaka (Oryzias latipes).

PubMed

Zha, Jinmiao; Wang, Zijian

2006-03-15

To develop the whole effluent toxicity testing methods (WET), embryo larval stage toxicity test using Japanese medaka (Oryzias latipes) was conducted to evaluate an effluent from a banknote printing plant (BPP). The method is based on acute toxicity using endpoint of 96-h larval morality and on chronic toxicity using endpoints such as the time to hatch, hatching success, deformity, growth rate, swim-up failure, accumulative mortality and sexual ratio. In test for 96-h larval mortality, LC50 (the concentration was lethal to 50% of newly hatching medaka larvae) was 72.9%. In chronic toxicity test, newly fertilized embryos (<5-h old) were exposed to 1%, 6.25%, 12.5%, 25%, 50% effluent concentrations and to 200 mug/l BPA in a 24-h static renewal system at 25+/-1 degrees C until 15 day post-hatch. The results showed that all chronic endpoints were significantly different from the control at 50% dilution (p < 0.01). Embryos began to show lesions on 4th day at higher concentrations (12.5%, 25%, 50% BPP effluent concentrations). Treatment group of 25% dilution showed delayed time to hatch. A reduction in body weight was observed at 25% dilutions for males and females, respectively. Deformities were observed in newly hatched larvae at 25% and 50% BPP effluent concentrations. At 25% dilution, sex ratio of larvae was alternated and there was feminization phenomenon. We conclude that embryo larval stage test using medaka is feasible to evaluate both acute and chronic toxicities and potential endocrine disrupting activity of industrial effluents.

DNA-based methodologies for the quantification of live and dead cells in formulated biocontrol products based on Pantoea agglomerans CPA-2.

PubMed

Soto-Muñoz, Lourdes; Torres, Rosario; Usall, Josep; Viñas, Inmaculada; Solsona, Cristina; Teixidó, Neus

2015-10-01

Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) against the major postharvest pathogens present on pome and citrus fruits. Dehydration, such as freeze-drying, spray-drying and fluidized bed drying is one of the best ways to formulate BCAs. In this work, the survival of CPA-2 cells after formulation was determined by dilution plating and molecular methods as qPCR alone and combined with a sample pretreatment with a propidium monoazide dye (PMA-qPCR) and they were used to calculate treatment concentrations in efficacy trials on postharvest oranges. Furthermore, no significant differences in CPA-2 survival were observed as determined by dilution plating and PMA-qPCR after both the freeze drying and fluidized bed drying processes; however, an interesting significant difference was observed in the spray dried product comparing all quantitative methods. A difference of 0.48 and 2.17 log10 CFU or cells g/dw was observed among PMA-qPCR with qPCR and dilution plating, respectively. According to our study, dilution plating was shown to be an unreliable tool for monitoring the survival of CPA-2 after spray drying. In contrast, the combination of PMA and qPCR enabled a quick and unequivocal methodology to enumerate viable and VBNC CPA-2 cells under stress-dried conditions. Efficacy trials showed that, after 3 days, spray drying formulation rehydrated with 10% non-fat skimmed milk (NFSM) was as effective as fresh cells to control Penicillium digitatum in oranges. Copyright © 2015 Elsevier B.V. All rights reserved.

Determination of calcium, magnesium and zinc in lubricating oils by flame atomic absorption spectrometry using a three-component solution.

PubMed

Zmozinski, Ariane V; de Jesus, Alexandre; Vale, Maria G R; Silva, Márcia M

2010-12-15

Lubricating oils are used to decrease wear and friction of movable parts of engines and turbines, being in that way essential for the performance and the increase of that equipment lifespan. The presence of some metals shows the addition of specific additives such as detergents, dispersals and antioxidants that improve the performance of these lubricants. In this work, a method for determination of calcium, magnesium and zinc in lubricating oil by flame atomic absorption spectrometry (F AAS) was developed. The samples were diluted with a small quantity of aviation kerosene (AVK), n-propanol and water to form a three-component solution before its introduction in the F AAS. Aqueous inorganic standards diluted in the same way have been used for calibration. To assess the accuracy of the new method, it was compared with ABNT NBR 14066 standard method, which consists in diluting the sample with AVK and in quantification by F AAS. Two other validating methods have also been used: the acid digestion and the certified reference material NIST (SRM 1084a). The proposed method provides the following advantages in relation to the standard method: significant reduction of the use of AVK, higher stability of the analytes in the medium and application of aqueous inorganic standards for calibration. The limits of detection for calcium, magnesium and zinc were 1.3 μg g(-1), 0.052 μg g(-1) and 0.41 μg g(-1), respectively. Concentrations of calcium, magnesium and zinc in six different samples obtained by the developed method did not differ significantly from the results obtained by the reference methods at the 95% confidence level (Student's t-test and ANOVA). Therefore, the proposed method becomes an efficient alternative for determination of metals in lubricating oil. Copyright © 2010 Elsevier B.V. All rights reserved.

Helicity moduli of three-dimensional dilute XY models

NASA Astrophysics Data System (ADS)

Garg, Anupam; Pandit, Rahul; Solla, Sara A.; Ebner, C.

1984-07-01

The helicity moduli of various dilute, classical XY models on three-dimensional lattices are studied with a view to understanding some aspects of the superfluidity of 4He in Vycor glass. A spinwave calculation is used to obtain the low-temperature helicity modulus of a regularly-diluted XY model. A similar calculation is performed for the randomly bond-diluted and site-diluted XY models in the limit of low dilution. A Monte Carlo simulation is used to obtain the helicity modulus of the randomly bond-diluted XY model over a wide range of temperature and dilution. It is found that the randomly diluted models do agree and the regularly diluted model does not agree with certain experimentally found features of the variation in superfluid fraction with coverage of 4He in Vycor glass.

Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction

PubMed Central

Mohammadi, Samira; Esfahani, Bahram Nasr; Moghim, Sharareh; Mirhendi, Hossein; Zaniani, Fatemeh Riyahi; Safaei, Hajieh Ghasemian; Fazeli, Hossein; Salehi, Mahshid

2017-01-01

Background: Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. Materials and Methods: The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples. Results: The CTAB method showed more positive results at 1:10–1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. Conclusions: According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor. PMID:29279831

METHOD FOR PREPARING NORMORPHINE

DOEpatents

Rapoport, H.; Look, M.

1959-06-01

An improved method is presented for producing normorphine from morphine. Morphine as the starting material is acetylated by treatment with acetylating agents to produce di-acetyl morphine (heroin). The acetylated compound is reacted with cyanating agents to produce di-acetyl-cyanonormorphine (cyanonorheroin). The di-acetyl-cyanonormorphine compound is then treated in accordance with the improved hydrolysis reactions of the present invention in which concentrated hydrochloric acid is employed for a limited time period to hydrolyze the acetyl group therefrom forming cyanonormorphine. Subsequently, the reaction mixture is diluted and hydrolysis of the cyano groups from the cyanonormorphine is effected with a longer contact time with dilute hydrochloric acid thereby producing normorphine. A high over-all conversion and production of a high purity product which may be radioactlvely labeled, if desired, is obtained by operation of the process.

Adaptation of the Sensititre broth microdilution technique to antimicrobial susceptibility testing of Mycoplasma hyopneumoniae.

PubMed

Tanner, A C; Erickson, B Z; Ross, R F

1993-09-01

A broth microdilution technique is described for determining the antimicrobial susceptibility of Mycoplasma hyopneumoniae, using commercially prepared Sensititre plates. Twenty-five field isolates and two reference strains (J & 232), were tested against seven antimicrobials. Field isolates were tested in duplicate and reference strains, four times to estimate reproducibility. Ninety-seven percent of the duplicate MIC results for the field isolates were in agreement, or within one log2 dilution. Similar results were obtained with the reference strains. The isolates were susceptible to lincomycin-spectinomycin, tylosin and oxytetracycline or resistant to amoxycillin, apramycin and erythromycin. Susceptibility to furaltadone varied. This method retains the accuracy and reproducibility of broth MIC determinations, while avoiding the lengthy preparation of antimicrobial dilutions normally associated with more traditional methods.

Bias due to Preanalytical Dilution of Rodent Serum for Biochemical Analysis on the Siemens Dimension Xpand Plus

PubMed Central

Johns, Jennifer L.; Moorhead, Kaitlin A.; Hu, Jing; Moorhead, Roberta C.

2018-01-01

Clinical pathology testing of rodents is often challenging due to insufficient sample volume. One solution in clinical veterinary and exploratory research environments is dilution of samples prior to analysis. However, published information on the impact of preanalytical sample dilution on rodent biochemical data is incomplete. The objective of this study was to evaluate the effects of preanalytical sample dilution on biochemical analysis of mouse and rat serum samples utilizing the Siemens Dimension Xpand Plus. Rats were obtained from end of study research projects. Mice were obtained from sentinel testing programs. For both, whole blood was collected via terminal cardiocentesis into empty tubes and serum was harvested. Biochemical parameters were measured on fresh and thawed frozen samples run straight and at dilution factors 2–10. Dilutions were performed manually, utilizing either ultrapure water or enzyme diluent per manufacturer recommendations. All diluted samples were generated directly from the undiluted sample. Preanalytical dilution caused clinically unacceptable bias in most analytes at dilution factors four and above. Dilution-induced bias in total calcium, creatinine, total bilirubin, and uric acid was considered unacceptable with any degree of dilution, based on the more conservative of two definitions of acceptability. Dilution often caused electrolyte values to fall below assay range precluding evaluation of bias. Dilution-induced bias occurred in most biochemical parameters to varying degrees and may render dilution unacceptable in the exploratory research and clinical veterinary environments. Additionally, differences between results obtained at different dilution factors may confound statistical comparisons in research settings. Comparison of data obtained at a single dilution factor is highly recommended. PMID:29497614

Synovial Fluid Response to Extensional Flow: Effects of Dilution and Intermolecular Interactions

PubMed Central

Haward, Simon J.

2014-01-01

In this study, a microfluidic cross-slot device is used to examine the extensional flow response of diluted porcine synovial fluid (PSF) samples using flow-induced birefringence (FIB) measurements. The PSF sample is diluted to 10× 20× and 30× its original mass in a phosphate-buffered saline and its FIB response measured as a function of the strain rate at the stagnation point of the cross-slots. Equivalent experiments are also carried out using trypsin-treated PSF (t-PSF) in which the protein content is digested away using an enzyme. The results show that, at the synovial fluid concentrations tested, the protein content plays a negligible role in either the fluid's bulk shear or extensional flow behaviour. This helps support the validity of the analysis of synovial fluid HA content, either by microfluidic or by other techniques where the synovial fluid is first diluted, and suggests that the HA and protein content in synovial fluid must be higher than a certain minimum threshold concentration before HA-protein or protein-protein interactions become significant. However a systematic shift in the FIB response as the PSF and t-PSF samples are progressively diluted indicates that HA-HA interactions remain significant at the concentrations tested. These interactions influence FIB-derived macromolecular parameters such as the relaxation time and the molecular weight distribution and therefore must be minimized for the best validity of this method as an analytical technique, in which non-interaction between molecules is assumed. PMID:24651529

Synovial fluid response to extensional flow: effects of dilution and intermolecular interactions.

PubMed

Haward, Simon J

2014-01-01

In this study, a microfluidic cross-slot device is used to examine the extensional flow response of diluted porcine synovial fluid (PSF) samples using flow-induced birefringence (FIB) measurements. The PSF sample is diluted to 10× 20× and 30× its original mass in a phosphate-buffered saline and its FIB response measured as a function of the strain rate at the stagnation point of the cross-slots. Equivalent experiments are also carried out using trypsin-treated PSF (t-PSF) in which the protein content is digested away using an enzyme. The results show that, at the synovial fluid concentrations tested, the protein content plays a negligible role in either the fluid's bulk shear or extensional flow behaviour. This helps support the validity of the analysis of synovial fluid HA content, either by microfluidic or by other techniques where the synovial fluid is first diluted, and suggests that the HA and protein content in synovial fluid must be higher than a certain minimum threshold concentration before HA-protein or protein-protein interactions become significant. However a systematic shift in the FIB response as the PSF and t-PSF samples are progressively diluted indicates that HA-HA interactions remain significant at the concentrations tested. These interactions influence FIB-derived macromolecular parameters such as the relaxation time and the molecular weight distribution and therefore must be minimized for the best validity of this method as an analytical technique, in which non-interaction between molecules is assumed.

Chemical Characterization of an Envelope A Sample from Hanford Tank 241-AN-103

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

Hay, M.S.

2000-08-23

A whole tank composite sample from Hanford waste tank 241-AN-103 was received at the Savannah River Technology Center (SRTC) and chemically characterized. Prior to characterization the sample was diluted to {approximately}5 M sodium concentration. The filtered supernatant liquid, the total dried solids of the diluted sample, and the washed insoluble solids obtained from filtration of the diluted sample were analyzed. A mass balance calculation of the three fractions of the sample analyzed indicate the analytical results appear relatively self-consistent for major components of the sample. However, some inconsistency was observed between results where more than one method of determination wasmore » employed and for species present in low concentrations. A direct comparison to previous analyses of material from tank 241-AN-103 was not possible due to unavailability of data for diluted samples of tank 241-AN-103 whole tank composites. However, the analytical data for other types of samples from 241-AN-103 we re mathematically diluted and compare reasonably with the current results. Although the segments of the core samples used to prepare the sample received at SRTC were combined in an attempt to produce a whole tank composite, determination of how well the results of the current analysis represent the actual composition of the Hanford waste tank 241-AN-103 remains problematic due to the small sample size and the large size of the non-homogenized waste tank.« less

Use of vitamin B12 in joint lavage for determination of dilution factors of canine synovial fluid.

PubMed

de Bruin, Tanya; de Rooster, Hilde; van Bree, Henri; Cox, Eric

2005-11-01

To test a modified saline (0.9% NaCl) solution joint washing (lavage) technique that includes the use of vitamin B12 as an internal marker for the evaluation of synovial fluid dilution in lavage samples from canine joints. 9 plasma samples obtained from blood samples of 9 healthy dogs and 9 synovial fluid samples aspirated from stifle joints of 9 cadaveric dogs. Photometric absorbances of 25% vitamin B12 solution, canine synovial fluid, and canine plasma were measured in a spectrophotometer to establish an optimal wavelength for analysis. Canine synovial fluid and plasma samples were mixed with the 25% vitamin B12 solution to obtain 1%, 3%, 5%, 10%, 20%, and 50% solutions of synovial fluid or plasma. Diluted synovial fluid and plasma samples were used to simulate joint lavage samples and to examine the possible interference of these substances (synovial fluid or plasma) with the absorbance of the 25% vitamin B12 solution in photometric analysis. The optimal wavelength was found to be at 550 nm. Canine synovial fluid and plasma samples did not interfere with the absorbance measurements of the 25% vitamin B12 solution up to a 50% dilution of plasma or synovial fluid. The modified saline solution joint lavage method with the use of a 25% vitamin B12 solution as an internal standard provides an accurate and reliable technique for the evaluation of synovial fluid dilution in lavage samples from canine joints.

Body fat of stock-type horses predicted by rump fat thickness and deuterium oxide dilution and validated by near-infrared spectroscopy of dissected tissues.

PubMed

Ferjak, E N; Cavinder, C A; Burnett, D D; Argo, C Mc; Dinh, T T N

2017-10-01

Body condition score and percent body fat (BF; %) of horses are positively correlated with reproductive efficiency and are indicative of metabolic issues. However, BF in horses may be poorly predicted because current procedures are either subjective or dependent on one anatomical location. Therefore, the objectives of the current study were to compare 2 methods of predicting BF using rump fat thickness (RFT) and deuterium oxide (DO) dilution with actual tissue fat analysis by near-infrared spectroscopy (NIRS) in stock-type horses and to identify the relationship between BF and BCS. Twenty-four stock-type horses were selected to be humanely euthanized based on 3 primary criteria: geriatric, crippled, and/or unsafe. Approximately 20 h before slaughter, horses were weighed and BCS assessed to be 1 ( = 1; 433 kg), 2 ( = 1; 415 kg), 3 ( = 1; 376 kg), 4 ( = 7; 468 ± 13 kg), 5 ( = 10; 455 ± 11 kg), and 6 ( = 4; 493 ± 12 kg) and RFT was measured using ultrasonography. Blood samples were collected immediately before and 4 h after DO infusion (0.12 g/kg BW). Deuterium oxide concentration of plasma was determined by gas isotope ratio mass spectrometry. Horses were housed in a dry lot overnight before being individually sedated (1.1 mg xylazine/kg BW) and anesthetized using a jugular venipuncture (2.2 mg ketamine/kg BW), and potassium chloride (KCl) solution was administered to cease cardiac function before exsanguination. After euthanasia, horse carcasses were processed and dissected and tissues were collected for NIRS analysis. Body fat predicted by DO dilution was correlated with BF measured by NIRS analysis on various weight bases ( = 0.76 to 0.81, < 0.001), whereas no correlation was observed for BF predicted by RFT. A paired -test indicated that both DO dilution and RFT overestimated BF by 2.48 to 3.26% ( < 0.001) and 5.81 to 6.59% ( < 0.001), respectively, compared with NIRS analysis. Body condition scores affected BF measured by NIRS analysis ( ≤ 0.038) and predicted by DO dilution ( < 0.001) and RFT ( = 0.042). The current study provided evidence that DO dilution was a better prediction method for BF than RFT and that BCS may serve as a BF predictor in stock-type horses.

Quantifying Trophic and Demographic Rates of Plankton-Rich Layers in East Sound, Orcas Island, Washington

DTIC Science & Technology

2006-01-01

are used, two independent methods to measure primary productivity and the dilution method to measure heterotrophic protists grazing rate. In...dinoflagellates, diatoms, and other protist plankton. Limnology and Oceanography 45: 569-579 Menden-Deuer S, EJ Lessard & J Satterberg. 2001

Platelet counting using the Coulter electronic counter

PubMed Central

Eggleton, M. J.; Sharp, A. A.

1963-01-01

A method for counting platelets in dilutions of platelet-rich plasm using the Coulter electronic counter is described.1 The results obtained show that such platelet counts are at least as accurate as the best methods of visual counting. The various technical difficulties encountered are discussed. PMID:16811002

In vitro effect of amoxicillin and clarithromycin on the 3’ region of cagA gene in Helicobacter pylori isolates

PubMed Central

Bustamante-Rengifo, Javier Andrés; Matta, Andrés Januer; Pazos, Alvaro; Bravo, Luis Eduardo

2013-01-01

AIM: To evaluate the in vitro effect of amoxicillin and clarithromycin on the cag pathogenicity island (cag PAI). METHODS: One hundred and forty-nine clinical isolates of Helicobacter pylori (H. pylori) cultured from gastric biopsies from 206 Colombian patients with dyspeptic symptoms from a high-risk area for gastric cancer were included as study material. Antimicrobial susceptibility was determined by the agar dilution method. Resistant isolates at baseline and in amoxicillin and clarithromycin serial dilutions were subjected to genotyping (cagA, vacA alleles s and m), Glu-Pro-Ile-Tyr-Ala (EPIYA) polymerase chain reaction and random amplified polymorphic DNA (RAPD). Images of the RAPD amplicons were analyzed by Gel-Pro Analyzer 4.5 program. Cluster analyses was done using SPSS 15.0 statistical package, where each of the fingerprint bands were denoted as variables. Dendrograms were designed by following Ward’s clustering method and the estimation of distances between each pair of H. pylori isolates was calculated with the squared Euclidean distance. RESULTS: Resistance rates were 4% for amoxicillin and 2.7% for clarithromycin with 2% double resistances. Genotyping evidenced a high prevalence of the genotype cagA-positive/vacA s1m1. The 3’ region of cagA gene was successfully amplified in 92.3% (12/13) of the baseline resistant isolates and in 60% (36/60) of the resistant isolates growing in antibiotic dilutions. Upon observing the distribution of the number of EPIYA repetitions in each dilution with respect to baseline isolates, it was found that in 61.5% (8/13) of the baseline isolates, a change in the number of EPIYA repetitions lowered antibiotic pressure. The gain and loss of EPIYA motifs resulted in a diversity of H. pylori subclones after bacterial adjustment to changing conditions product of antibiotic pressure. RAPD PCR evidenced the close clonal relationship between baseline isolates and isolates growing in antibiotic dilutions. CONCLUSION: Antibiotic pressure does not induce loss of the cag pathogenicity island, but it can lead - in most cases - to genetic rearrangements within the 3’ region cagA of the founding bacteria that can affect the level of tyrosine phosphorylation impacting on its cellular effects and lead to divergence of cagA-positive subclones. PMID:24106405

Iodine retention during evaporative volume reduction

DOEpatents

Godbee, H.W.; Cathers, G.I.; Blanco, R.E.

1975-11-18

An improved method for retaining radioactive iodine in aqueous waste solutions during volume reduction is disclosed. The method applies to evaporative volume reduction processes whereby the decontaminated (evaporated) water can be returned safely to the environment. The method generally comprises isotopically diluting the waste solution with a nonradioactive iodide and maintaining the solution at a high pH during evaporation.

Stable isotope dilution assay (SIDA) and HS-SPME-GCMS quantification of key aroma volatiles for fruit and sap of Australian mango cultivars.

PubMed

San, Anh T; Joyce, Daryl C; Hofman, Peter J; Macnish, Andrew J; Webb, Richard I; Matovic, Nicolas J; Williams, Craig M; De Voss, James J; Wong, Siew H; Smyth, Heather E

2017-04-15

Reported herein is a high throughput method to quantify in a single analysis the key volatiles that contribute to the aroma of commercially significant mango cultivars grown in Australia. The method constitutes stable isotope dilution analysis (SIDA) in conjunction with headspace (HS) solid-phase microextraction (SPME) coupled with gas-chromatography mass spectrometry (GCMS). Deuterium labelled analogues of the target analytes were either purchased commercially or synthesised for use as internal standards. Seven volatiles, hexanal, 3-carene, α-terpinene, p-cymene, limonene, α-terpinolene and ethyl octanoate, were targeted. The resulting calibration functions had determination coefficients (R 2 ) ranging from 0.93775 to 0.99741. High recovery efficiencies for spiked mango samples were also achieved. The method was applied to identify the key aroma volatile compounds produced by 'Kensington Pride' and 'B74' mango fruit and by 'Honey Gold' mango sap. This method represents a marked improvement over current methods for detecting and measuring concentrations of mango fruit and sap volatiles. Copyright © 2016 Elsevier Ltd. All rights reserved.

The immediate impact of semen diluent and rate of dilution on the sperm quality index, ATP utilization, gas exchange, and ionic balance of broiler breeder sperm.

PubMed

Parker, H M; McDaniel, C D

2006-01-01

The sperm quality index (SQI) is a tool used to predict overall rooster semen quality, fertility, and hatchability. However, semen must be diluted before SQI analysis, and research has shown that the SQI is most predictive of fertility at lower semen dilutions. Therefore, the present study was undertaken to determine why the SQI is not as predictive of fertility at higher semen dilutions and whether semen diluent type alters the SQI, adenosine triphosphate (ATP) utilization, gas exchange, and ionic balance of broiler breeder sperm. Semen was diluted with saline, seminal plasma, or minimum essential medium (MEM) from 2- to 200-fold. The following parameters were measured for each diluent type at each dilution: SQI, ATP, Na+, Ca2+, K+, Cl-, CO2, and O2. To examine the rate of sperm motility, the SQI was expressed as SQI/million sperm per mL (SQI/sperm). There was an interaction between diluent type and dilution for the SQI, SQI/sperm, CO2 generated, O2 used, as well as Na+, Ca2+, and K+ internalization. For sperm diluted with saline, the SQI declined more rapidly with increasing dilution. However, SQI/sperm increased rapidly when semen was diluted with MEM or SP. Sperm diluted in SP used ATP with increasing dilution whereas sperm diluted with saline and MEM generated ATP. Neat semen contained no free O2; however, each diluent type contained abundant O2 resulting in more O2 available as semen was diluted. Sperm diluted in SP produced more CO2 and used more O2 than semen diluted in saline or MEM. For SQI/sperm, ATP and CO2 generated, as well as Na+ and Ca2+ internalization, differences between diluent types occurred when semen was diluted 50-fold and greater. In conclusion, it appears that sperm motility, ATP utilization, gas exchange, and ionic balance are altered by diluent type and rate of dilution. These alterations in semen quality are exacerbated at semen dilutions of 50-fold and greater yielding an SQI that is not indicative of sperm motility or fertility.

Comparison of plethysmographic and helium dilution lung volumes in patients with a giant emphysematous bulla as selection criteria for endobronchial valve implant.

PubMed

Fiorelli, Alfonso; Scaramuzzi, Roberto; Pierdiluca, Matteo; Frongillo, Elisabetta; Messina, Gaetana; Serra, Nicola; De Felice, Alberto; Santini, Mario

2017-09-01

To assess whether the difference in lung volume measured with plethysmography and with the helium dilution technique could differentiate an open from a closed bulla in patients with a giant emphysematous bulla and could be used as a selection criterion for the positioning of an endobronchial valve. We reviewed the data of 27 consecutive patients with a giant emphysematous bulla undergoing treatment with an endobronchial valve. In addition to standard functional and radiological examinations, total lung capacity and residual volume were measured with the plethysmographic and helium dilution technique. We divided the patients into 2 groups, the collapse or the no-collapse group, depending on whether the bulla collapsed or not after the valves were put in position. We statistically evaluated the intergroup differences in lung volume and outcome. In the no-collapse group (n = 6), the baseline plethysmographic values were significantly higher than the helium dilution volumes, including total lung capacity (188 ± 14 vs 145 ± 13, P = 0.0007) and residual volume (156 ± 156 vs 115 ± 15, P = 0.001). In the collapse group, there was no significant difference in lung volumes measured with the 2 methods. A difference in total lung capacity of ≤ 13% and in residual volume of ≤ 25% measured with the 2 methods predicted the collapse of the bulla with a success rate of 83% and 84%, respectively. Only the collapse group showed significant improvement in functional data. Similar values in lung volumes measured with the 2 methods support the hypothesis that the bulla communicates with the airway (open bulla) and thus is likely to collapse when the endobronchial valve is implanted. Further studies are needed to validate our model. © The Author 2017. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

Flexible nano- and microliter injections on a single liquid chromatography-mass spectrometry system: Minimizing sample preparation and maximizing linear dynamic range.

PubMed

Lubin, Arnaud; Sheng, Sheng; Cabooter, Deirdre; Augustijns, Patrick; Cuyckens, Filip

2017-11-17

Lack of knowledge on the expected concentration range or insufficient linear dynamic range of the analytical method applied are common challenges for the analytical scientist. Samples that are above the upper limit of quantification are typically diluted and reanalyzed. The analysis of undiluted highly concentrated samples can cause contamination of the system, while the dilution step is time consuming and as the case for any sample preparation step, also potentially leads to precipitation, adsorption or degradation of the analytes. Copyright © 2017 Elsevier B.V. All rights reserved.

Fire Suppression M and S Validation (Status and Challenges), Systems Fire Protection Information Exchange

DTIC Science & Technology

2015-10-14

rate Kinetics •14 Species & 12 reactionsCombustion Model •Participating Media Discrete Ordinate Method •WSG model for CO2, H2O and SootRadiation Model...Inhibition of JP-8 Combustion Physical Acting Agents • Dilute heat • Dilute reactants Ex: water, nitrogen Chemical Acting Agents • Reduce flame...Release; distribution is unlimited 5 Overview of Reduced Kinetics Scheme for FM200 • R1: JP-8 + O2 => CO + CO2 + H2O • R2: CO + O2 <=> CO2 • R3: HFP + JP-8

Diluted magnetic oxides

NASA Astrophysics Data System (ADS)

Li, XiaoLi; Qi, ShiFei; Jiang, FengXian; Quan, ZhiYong; Xu, XiaoHong

2013-01-01

In this review, we review the progress of research on ZnO- and In2O3-based diluted magnetic oxides (DMOs). Firstly, we present the preparation and characterization of DMOs. The former includes the preparation methods and conditions, and the latter includes the characterization techniques for measuring microstructures. Secondly, we introduce the magnetic and transport properties of DMOs, as well as the relationship between them. Thirdly, the origin and mechanism of the ferromagnetism are discussed. Fourthly, we introduce other related work, including computational work and pertinent heterogeneous structures, such as multilayers and magnetic tunnel junctions. Finally, we provide an overview and outlook for DMOs.

Berezinskii-Kosterlitz-Thouless phase transition for the dilute planar rotator model on a triangular lattice

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

Sun Yunzhou; Yi Lin; Wysin, G. M.

2008-10-15

The Berezinskii-Kosterlitz-Thouless (BKT) phase transition for the dilute planar rotator model on a triangular lattice is studied by using a hybrid Monte Carlo method. The phase-transition temperatures for different nonmagnetic impurity densities are obtained by three approaches: finite-size scaling of plane magnetic susceptibility, helicity modulus, and Binder's fourth cumulant. It is found that the phase-transition temperature decreases with increasing impurity density {rho} and the BKT phase transition vanishes when the magnetic occupancy falls to the site percolation threshold: 1-{rho}{sub c}=p{sub c}=0.5.

Method and apparatus for assaying wood pulp fibers

DOEpatents

Gustafson, Richard [Bellevue, WA; Callis, James B [Seattle, WA; Mathews, Jeffrey D [Neenah, WI; Robinson, John [Issaquah, WA; Bruckner, Carsten A [San Mateo, CA; Suvamakich, Kuntinee [Seattle, WA

2009-05-26

Paper pulp is added to a stain solution. The stain solution and pulp fibers are mixed to form a slurry. Samples are removed from the slurry and are admixed with dilution water and a bleach. Then, the fibers are moved into a flow cell where they are subjected to a light source adapted to stimulate fluorescence from the stained pulp fiber. Before the fiber slurry enters the flow cell it is mixed with a dilution water of bleach to reduce background fluorescence. The fluorescent light is collimated and directed through a dichroic filter onto a fluorescence splitting dichroic filter.

Automated measurement of respiratory gas exchange by an inert gas dilution technique

NASA Technical Reports Server (NTRS)

Sawin, C. F.; Rummel, J. A.; Michel, E. L.

1974-01-01

A respiratory gas analyzer (RGA) has been developed wherein a mass spectrometer is the sole transducer required for measurement of respiratory gas exchange. The mass spectrometer maintains all signals in absolute phase relationships, precluding the need to synchronize flow and gas composition as required in other systems. The RGA system was evaluated by comparison with the Douglas bag technique. The RGA system established the feasibility of the inert gas dilution method for measuring breath-by-breath respiratory gas exchange. This breath-by-breath analytical capability permits detailed study of transient respiratory responses to exercise.

Recovery of organic acids

DOEpatents

Verser, Dan W.; Eggeman, Timothy J.

2009-10-13

A method is disclosed for the recovery of an organic acid from a dilute salt solution in which the cation of the salt forms an insoluble carbonate salt. A tertiary amine and CO.sub.2 are introduced to the solution to form the insoluble carbonate salt and a complex between the acid and an amine. A water immiscible solvent, such as an alcohol, is added to extract the acid/amine complex from the dilute salt solution to a reaction phase. The reaction phase is continuously dried and a product between the acid and the solvent, such as an ester, is formed.

Recovery of organic acids

DOEpatents

Verser, Dan W [Menlo Park, CA; Eggeman, Timothy J [Lakewood, CO

2011-11-01

A method is disclosed for the recovery of an organic acid from a dilute salt solution in which the cation of the salt forms an insoluble carbonate salt. A tertiary amine and CO.sub.2 are introduced to the solution to form the insoluble carbonate salt and a complex between the acid and an amine. A water immiscible solvent, such as an alcohol, is added to extract the acid/amine complex from the dilute salt solution to a reaction phase. The reaction phase is continuously dried and a product between the acid and the solvent, such as an ester, is formed.

Convergent beam electron-diffraction investigation of lattice mismatch and static disorder in GaAs/GaAs1-xNx intercalated GaAs/GaAs1-xNx:H heterostructures

NASA Astrophysics Data System (ADS)

Frabboni, S.; Grillo, V.; Gazzadi, G. C.; Balboni, R.; Trotta, R.; Polimeni, A.; Capizzi, M.; Martelli, F.; Rubini, S.; Guzzinati, G.; Glas, F.

2012-09-01

Hydrogen incorporation in diluted nitride semiconductors dramatically modifies the electronic and structural properties of the crystal through the creation of nitrogen-hydrogen complexes. We report a convergent beam electron-diffraction characterization of diluted nitride semiconductor-heterostructures patterned at a sub-micron scale and selectively exposed to hydrogen. We present a method to determine separately perpendicular mismatch and static disorder in pristine and hydrogenated heterostructures. The roles of chemical composition and strain on static disorder have been separately assessed.

Preparation of chitosan/tripolyphosphate nanoparticles with highly tunable size and low polydispersity.

PubMed

Sawtarie, Nader; Cai, Yuhang; Lapitsky, Yakov

2017-09-01

Nanoparticles prepared through the ionotropic gelation of chitosan with tripolyphosphate (TPP) have been extensively studied as vehicles for drug and gene delivery. Though a number of these works have focused on preparing particles with narrow size distributions, the monodisperse particles produced by these methods have been limited to narrow size ranges (where the average particle size was not varied by more than twofold). Here we show how, by tuning the NaCl concentration in the parent chitosan and TPP solutions, low-polydispersity particles with z-average diameters ranging between roughly 100 and 900nm can be prepared. Further, we explore how the size of these particles depends on the method by which the TPP is mixed into the chitosan solution, specifically comparing: (1) single-shot mixing; (2) dropwise addition; and (3) a dilution technique, where chitosan and TPP are codissolved at a high (gelation-inhibiting) ionic strength and then diluted to lower ionic strengths to trigger gelation. Though the particle size increases sigmoidally with the NaCl concentration for all three mixing methods, the dilution method delivers the most uniform/gradual size increase - i.e., it provides the most precise control. Also investigated are the effects of mixture composition and mixing procedure on the particle yield. These reveal the particle yield to increase with the chitosan/TPP concentration, decrease with the NaCl concentration, and vary only weakly with the mixing protocol; thus, at elevated NaCl concentrations, it may be beneficial to increase chitosan and TPP concentrations to ensure high particle yields. Finally, possible pitfalls of the salt-assisted size control strategy (and their solutions) are discussed. Taken together, these findings provide a simple and reliable method for extensively tuning chitosan/TPP particle size while maintaining narrow size distributions. Copyright © 2017 Elsevier B.V. All rights reserved.

Determination of serum calcium levels by 42Ca isotope dilution inductively coupled plasma mass spectrometry.

PubMed

Han, Bingqing; Ge, Menglei; Zhao, Haijian; Yan, Ying; Zeng, Jie; Zhang, Tianjiao; Zhou, Weiyan; Zhang, Jiangtao; Wang, Jing; Zhang, Chuanbao

2017-11-27

Serum calcium level is an important clinical index that reflects pathophysiological states. However, detection accuracy in laboratory tests is not ideal; as such, a high accuracy method is needed. We developed a reference method for measuring serum calcium levels by isotope dilution inductively coupled plasma mass spectrometry (ID ICP-MS), using 42Ca as the enriched isotope. Serum was digested with 69% ultrapure nitric acid and diluted to a suitable concentration. The 44Ca/42Ca ratio was detected in H2 mode; spike concentration was calibrated by reverse IDMS using standard reference material (SRM) 3109a, and sample concentration was measured by a bracketing procedure. We compared the performance of ID ICP-MS with those of three other reference methods in China using the same serum and aqueous samples. The relative expanded uncertainty of the sample concentration was 0.414% (k=2). The range of repeatability (within-run imprecision), intermediate imprecision (between-run imprecision), and intra-laboratory imprecision were 0.12%-0.19%, 0.07%-0.09%, and 0.16%-0.17%, respectively, for two of the serum samples. SRM909bI, SRM909bII, SRM909c, and GBW09152 were found to be within the certified value interval, with mean relative bias values of 0.29%, -0.02%, 0.10%, and -0.19%, respectively. The range of recovery was 99.87%-100.37%. Results obtained by ID ICP-MS showed a better accuracy than and were highly correlated with those of other reference methods. ID ICP-MS is a simple and accurate candidate reference method for serum calcium measurement and can be used to establish and improve serum calcium reference system in China.

Measurement of fumonisins in corn with a fiber optic fluoroimmunosensor

NASA Astrophysics Data System (ADS)

Thompson, Vicki S.; Maragos, Chris M.

1997-05-01

A fiber-optic immunosensor was used to determine concentrations of the mycotoxin fumonisin B1(FB1) in both spiked and naturally contaminated corn samples. Samples were extracted with a mixture of methanol/water. Two methods were used to prepare the methanolic corn extracts before introduction to the immunosensor: (1) simple dilution of the methanolic corn extract; or (2) affinity column cleanup. The sensor displayed an IC50 of 70 ng FB1/mL when toxin was introduced in phosphate buffered saline. Simple dilution of methanolic corn extracts yielded an assay with an IC50 equivalent to 25 (mu) gFB1/g corn and a limit of detection of 3.2 (mu) g/g corn, while affinity cleanup of corn extracts yielded an assay with an IC50 of 5 (mu) gFB1/g corn and a limit of detection of 0.4 (mu) gFB1/g corn. The difference in sensitivity between the two cleanup techniques was due to concentration of fumonisins obtained from the affinity cleanup procedure. Naturally contaminated corn samples were also analyzed after either simple dilution or affinity column cleanup. For comparison the naturally contaminated corn samples were analyzed with an HPLC method after isolation of the fumonisins with strong anion exchange (SAX) solid phase extraction cartridges. The SAX/HPLC method and the immunosensor method agreed well except when large amounts of other fumonisins (i.e. fumonisin B2) were present. This was due in part to the cross-reactivity of the monoclonal antibody with other fumonisins. The immunosensor has the potential to screen individual corn samples for fumonisins within six minutes, and is among the fastest of the currently available FB1 detection methods.

Patch testing in non-immediate cutaneous adverse drug reactions: value of extemporaneous patch tests.

PubMed

Assier, Haudrey; Valeyrie-Allanore, Laurence; Gener, Gwendeline; Verlinde Carvalh, Muriel; Chosidow, Olivier; Wolkenstein, Pierre

2017-11-01

Patch testing following a standardized protocol is reliable for identifying the culprit drug in cutaneous adverse drug reactions (CADRs). However, these patch tests (PTs) require pharmaceutical material and staff, which are not always easily available. To evaluate an extemporaneous PT method in CADRs. We retrospectively analysed data for all patients referred to our department between March 2009 and June 2013 for patch testing after a non-immediate CADR. The patients who supplied their own suspected drugs were tested both with extemporaneous PTs and with conventional PTs. Extemporaneous PTs involved a nurse crushing and diluting the drug in pet. in a ratio of approximately one-third to two-thirds. Standardized PTs were performed according to guidelines, with commercial drugs diluted to 30% or with active ingredients diluted to 10%. We analysed the data for the two PT methods in terms of the number of positive test reactions, drugs tested, and type of CADR for patients in whom the two PT methods were used. In total, 75 of 156 patients underwent the two PT procedures, including 91 double tests. Overall, 21 tests gave positive reactions with the two methods, and 69 other tests gave negative results with the two methods. Our series yielded results similar to those of published series concerning the types of CADR and the drugs responsible. Our results suggest that, for CADRs, if a patient supplies a suspected drug but if the pharmaceutical material and staff are not available for conventional PTs, extemporaneous PTs performed by the nurse with the commercial drug used by the patient can be useful and reliable. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Source selection for cluster weak lensing measurements in the Hyper Suprime-Cam survey

NASA Astrophysics Data System (ADS)

Medezinski, Elinor; Oguri, Masamune; Nishizawa, Atsushi J.; Speagle, Joshua S.; Miyatake, Hironao; Umetsu, Keiichi; Leauthaud, Alexie; Murata, Ryoma; Mandelbaum, Rachel; Sifón, Cristóbal; Strauss, Michael A.; Huang, Song; Simet, Melanie; Okabe, Nobuhiro; Tanaka, Masayuki; Komiyama, Yutaka

2018-03-01

We present optimized source galaxy selection schemes for measuring cluster weak lensing (WL) mass profiles unaffected by cluster member dilution from the Subaru Hyper Suprime-Cam Strategic Survey Program (HSC-SSP). The ongoing HSC-SSP survey will uncover thousands of galaxy clusters to z ≲ 1.5. In deriving cluster masses via WL, a critical source of systematics is contamination and dilution of the lensing signal by cluster members, and by foreground galaxies whose photometric redshifts are biased. Using the first-year CAMIRA catalog of ˜900 clusters with richness larger than 20 found in ˜140 deg2 of HSC-SSP data, we devise and compare several source selection methods, including selection in color-color space (CC-cut), and selection of robust photometric redshifts by applying constraints on their cumulative probability distribution function (P-cut). We examine the dependence of the contamination on the chosen limits adopted for each method. Using the proper limits, these methods give mass profiles with minimal dilution in agreement with one another. We find that not adopting either the CC-cut or P-cut methods results in an underestimation of the total cluster mass (13% ± 4%) and the concentration of the profile (24% ± 11%). The level of cluster contamination can reach as high as ˜10% at R ≈ 0.24 Mpc/h for low-z clusters without cuts, while employing either the P-cut or CC-cut results in cluster contamination consistent with zero to within the 0.5% uncertainties. Our robust methods yield a ˜60 σ detection of the stacked CAMIRA surface mass density profile, with a mean mass of M200c = [1.67 ± 0.05(stat)] × 1014 M⊙/h.

Modified Sample Preparation Approach for the Determination of the Phenolic and Humic-Like Substances in Natural Organic Materials By the Folin Ciocalteu Method.

PubMed

Pontoni, Ludovico; Panico, Antonio; Matanò, Alessia; van Hullebusch, Eric D; Fabbricino, Massimiliano; Esposito, Giovanni; Pirozzi, Francesco

2017-12-06

A novel modification of the sample preparation procedure for the Folin-Ciocalteu colorimetric assay for the determination of total phenolic compounds in natural solid and semisolid organic materials (e.g., foods, organic solid waste, soils, plant tissues, agricultural residues, manure) is proposed. In this method, the sample is prepared by adding sodium sulfate as a solid diluting agent before homogenization. The method allows for the determination of total phenols (TP) in samples with high solids contents, and it provides good accuracy and reproducibility. Additionally, this method permits analyses of significant amounts of sample, which reduces problems related to heterogeneity. We applied this method to phenols-rich lignocellulosic and humic-like solids and semisolid samples, including rice straw (RS), peat-rich soil (PS), and food waste (FW). The TP concentrations measured with the solid dilution (SD) preparation were substantially higher (increases of 41.4%, 15.5%, and 59.4% in RS, PS and FW, respectively) than those obtained with the traditional method (solids suspended in water). These results showed that the traditional method underestimates the phenolic contents in the studied solids.

A novel reversed-phase HPLC method for the determination of urinary creatinine by pre-column derivatization with ethyl chloroformate: comparative studies with the standard Jaffé and isotope-dilution mass spectrometric assays.

PubMed

Leung, Elvis M K; Chan, Wan

2014-02-01

Creatinine is an important biomarker for renal function diagnosis and normalizing variations in urinary drug/metabolites concentration. Quantification of creatinine in biological fluids such as urine and plasma is important for clinical diagnosis as well as in biomonitoring programs and urinary metabolomics/metabonomics research. Current methods for creatinine determination either are nonselective or involve the use of expensive mass spectrometers. In this paper, a novel reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of creatinine of high hydrophilicity by pre-column derivatization with ethyl chloroformate is presented. N-Ethyloxycarbonylation of creatinine significantly enhanced the hydrophobicity of creatinine, facilitating its chromatographic retention as well as quantification by HPLC. Factors governing the derivatization reaction were studied and optimized. The developed method was validated and applied for the determination of creatinine in rat urine samples. Comparative studies with isotope-dilution mass spectrometric method revealed that the two methods do not yield systematic differences in creatinine concentrations, indicating the HPLC method is suitable for the determination of creatinine in urine samples.

Application and validation of isotope dilution method (IDM) for predicting bioavailability of hydrophobic organic contaminants in soil.

PubMed

Wang, Jie; Taylor, Allison; Schlenk, Daniel; Gan, Jay

2018-05-01

Risk assessment of hydrophobic organic contaminants (HOCs) using the total chemical concentration following exhaustive extraction may overestimate the actual availability of HOCs to non-target organisms. Existing methods for estimating HOC bioavailability in soil have various operational limitations. In this study, we explored the application of isotope dilution method (IDM) to quantify the accessible fraction (E) of DDTs and PCBs in both historically-contaminated and freshly-spiked soils. After addition of 13 C or deuterated analogues to a soil sample, the phase distribution of isotope-labeled and native chemicals reached an apparent equilibrium within 48 h of mixing. The derived E values in the three soils ranged from 0.19 to 0.82, depending on the soil properties and also the contact time of HOCs (i.e., aging). The isotope dilution method consistently predicted greater accumulation into earthworm (Eisenia fetida) than that by polyethylene (PE) or solid phase microextraction (SPME) sampler, likely because desorption in the gut enhanced bioavailability of soil-borne HOCs. A highly significant linear regression (R 2  = 0.91) was found between IDM and 24-h Tenax desorption, with a slope statistically identical to 1. The IDM-derived accessible concentration (C e ) was further shown to accurately predict tissue residues in earthworm exposed in the same soils. Given the relatively short duration and simple steps, IDM has the potential to be readily adopted for measuring HOC bioaccessibility in soil and for improving risk assessment and evaluation of remediation efficiency. Copyright © 2018 Elsevier Ltd. All rights reserved.

Flow cytometry as an improved method for the titration of Chlamydiaceae and other intracellular bacteria.

PubMed

Käser, T; Pasternak, J A; Hamonic, G; Rieder, M; Lai, K; Delgado-Ortega, M; Gerdts, V; Meurens, F

2016-05-01

Chlamydiaceae is a family of intracellular bacteria causing a range of diverse pathological outcomes. The most devastating human diseases are ocular infections with C. trachomatis leading to blindness and genital infections causing pelvic inflammatory disease with long-term sequelae including infertility and chronic pelvic pain. In order to enable the comparison of experiments between laboratories investigating host-chlamydia interactions, the infectious titer has to be determined. Titer determination of chlamydia is most commonly performed via microscopy of host cells infected with a serial dilution of chlamydia. However, other methods including fluorescent ELISpot (Fluorospot) and DNA Chip Scanning Technology have also been proposed to enumerate chlamydia-infected cells. For viruses, flow cytometry has been suggested as a superior alternative to standard titration methods. In this study we compared the use of flow cytometry with microscopy and Fluorospot for the titration of C. suis as a representative of other intracellular bacteria. Titer determination via Fluorospot was unreliable, while titration via microscopy led to a linear read-out range of 16 - 64 dilutions and moderate reproducibility with acceptable standard deviations within and between investigators. In contrast, flow cytometry had a vast linear read-out range of 1,024 dilutions and the lowest standard deviations given a basic training in these methods. In addition, flow cytometry was faster and material costs were lower compared to microscopy. Flow cytometry offers a fast, cheap, precise, and reproducible alternative for the titration of intracellular bacteria like C. suis. © 2016 International Society for Advancement of Cytometry. © 2016 International Society for Advancement of Cytometry.

Method to make accurate concentration and isotopic measurements for small gas samples

NASA Astrophysics Data System (ADS)

Palmer, M. R.; Wahl, E.; Cunningham, K. L.

2013-12-01

Carbon isotopic ratio measurements of CO2 and CH4 provide valuable insight into carbon cycle processes. However, many of these studies, like soil gas, soil flux, and water head space experiments, provide very small gas sample volumes, too small for direct measurement by current constant-flow Cavity Ring-Down (CRDS) isotopic analyzers. Previously, we addressed this issue by developing a sample introduction module which enabled the isotopic ratio measurement of 40ml samples or smaller. However, the system, called the Small Sample Isotope Module (SSIM), does dilute the sample during the delivery with inert carrier gas which causes a ~5% reduction in concentration. The isotopic ratio measurements are not affected by this small dilution, but researchers are naturally interested accurate concentration measurements. We present the accuracy and precision of a new method of using this delivery module which we call 'double injection.' Two portions of the 40ml of the sample (20ml each) are introduced to the analyzer, the first injection of which flushes out the diluting gas and the second injection is measured. The accuracy of this new method is demonstrated by comparing the concentration and isotopic ratio measurements for a gas sampled directly and that same gas measured through the SSIM. The data show that the CO2 concentration measurements were the same within instrument precision. The isotopic ratio precision (1σ) of repeated measurements was 0.16 permil for CO2 and 1.15 permil for CH4 at ambient concentrations. This new method provides a significant enhancement in the information provided by small samples.

a Borehole-Dilution Method for Quantifying Vertical Darcy Fluxes in the Hyporheic Zone

NASA Astrophysics Data System (ADS)

Augustine, S. D.; Annable, M. D.; Cho, J.

2017-12-01

The borehole dilution method has consistently and successfully been used for estimating local water fluxes, however, this method can be relatively labor intensive and expensive. The focus of this research is aimed at developing a low-cost, borehole dilution method for quantifying vertical water fluxes in the hyporheic zone at the surface-groundwater interface. This would allow for the deployment of multiple units within a targeted surface water body and thus produce high-resolution, spatially distributed data on the infiltration rates over a short period of time with minimal set-up requirements. The device consists of a 2-inch, inner diameter PVC pipe containing short, screened sections in its upper and lower segments. The working unit is driven into the sediment and acts as a continuous flow reactor creating a pathway between the subsurface pore-water and the overlying surface water where the presence of a hydraulic gradient facilitates vertical movement. We developed a simple electrode and tracer-injection system housed within the unit to inject and measure salt tracer concentrations at the desired intervals while monitoring and storing those measurements using open-source Arduino technology. Preliminary lab and field scale trials provided data that was fit to both zero and first order reaction rate functions for analysis. The field test was conducted over approximately one day within a wet retention basin. The initial results estimated a vertical Darcy flux of 113.5 cm/d. Additional testing over a range of expected Darcy fluxes will be presented along with an evaluation considering enhanced water flow due to the high hydraulic conductivity of the device.

Specialized physiological studies in support of manned space flight

NASA Technical Reports Server (NTRS)

Luft, U. C.

1974-01-01

Research in aerospace medicine is presented. The reports discussed include: circulatory and respiratory transients during and after orthostasis and the effects of beta adrenergic blockade; the determination of total body water by an ethanol dilution method; and increased total respiratory conductance breathing 100% oxygen (forced oscillation method).

21 CFR 177.1950 - Vinyl chloride-ethylene copolymers.

Code of Federal Regulations, 2013 CFR

2013-04-01

... deciliter per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution... vinyl chloride-ethylene copol-ymer per 100 grams of sample tested as determined from the organic... using duplicate blanks. Approximately 400 grams of sample (accurately weighed) shall be placed in a 2...

21 CFR 177.1950 - Vinyl chloride-ethylene copolymers.

Code of Federal Regulations, 2012 CFR

2012-04-01

... deciliter per gram as determined by ASTM method D1243-79, “Standard Test Method for Dilute Solution... vinyl chloride-ethylene copol-ymer per 100 grams of sample tested as determined from the organic... using duplicate blanks. Approximately 400 grams of sample (accurately weighed) shall be placed in a 2...

A SEDIMENT TOXICITY METHOD USING LEMNA MINOR, DUCKWEED

EPA Science Inventory

We developed a Lemna minor sediment toxicity test method to assess sediment contaminants which may affect plants. This 96-hour test used 15 ml of sediment and 2 ml of overlying water which was renewed after 48 hours. Sand was used as the control sediment and also to dilute test ...

Analysis of thyroid hormones in biological samples using stable isotope dilution liquid chromatography-tandem mass spectrometry

EPA Science Inventory

This poster presentation will describe analytical chemistry methods for measuring thyroid hormones and related precursors and metabolites in very small tissue or plasma samples. These methods are amenable to measure thyroid hormones in amphibian tadpoles or small mammals used as ...

Thermodynamics of Dilute Solutions.

ERIC Educational Resources Information Center

Jancso, Gabor; Fenby, David V.

1983-01-01

Discusses principles and definitions related to the thermodynamics of dilute solutions. Topics considered include dilute solution, Gibbs-Duhem equation, reference systems (pure gases and gaseous mixtures, liquid mixtures, dilute solutions), real dilute solutions (focusing on solute and solvent), terminology, standard states, and reference systems.…

Analyses of turbulent flow fields and aerosol dynamics of diesel engine exhaust inside two dilution sampling tunnels using the CTAG model.

PubMed

Wang, Yan Jason; Yang, Bo; Lipsky, Eric M; Robinson, Allen L; Zhang, K Max

2013-01-15

Experimental results from laboratory emission testing have indicated that particulate emission measurements are sensitive to the dilution process of exhaust using fabricated dilution systems. In this paper, we first categorize the dilution parameters into two groups: (1) aerodynamics (e.g., mixing types, mixing enhancers, dilution ratios, residence time); and (2) mixture properties (e.g., temperature, relative humidity, particle size distributions of both raw exhaust and dilution gas). Then we employ the Comprehensive Turbulent Aerosol Dynamics and Gas Chemistry (CTAG) model to investigate the effects of those parameters on a set of particulate emission measurements comparing two dilution tunnels, i.e., a T-mixing lab dilution tunnel and a portable field dilution tunnel with a type of coaxial mixing. The turbulent flow fields and aerosol dynamics of particles are simulated inside two dilution tunnels. Particle size distributions under various dilution conditions predicted by CTAG are evaluated against the experimental data. It is found that in the area adjacent to the injection of exhaust, turbulence plays a crucial role in mixing the exhaust with the dilution air, and the strength of nucleation dominates the level of particle number concentrations. Further downstream, nucleation terminates and the growth of particles by condensation and coagulation continues. Sensitivity studies reveal that a potential unifying parameter for aerodynamics, i.e., the dilution rate of exhaust, plays an important role in new particle formation. The T-mixing lab tunnel tends to favor the nucleation due to a larger dilution rate of the exhaust than the coaxial mixing field tunnel. Our study indicates that numerical simulation tools can be potentially utilized to develop strategies to reduce the uncertainties associated with dilution samplings of emission sources.

Compatibility of an Ultraselective Microcatheter and Epirubicin Loaded 300–500-μm DC Bead in Ex Vivo Study

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

Fukuoka, Yasushi, E-mail: miyano.y.fukuoka@gmail.com; Tanaka, Toshihiro, E-mail: toshihir@bf6.so-net.ne.jp; Nishiofuku, Hideyuki, E-mail: nishiofukuhideyuki@yahoo.co.jp

PurposeThe purpose of this study is to examine whether epirubicin loaded DC Bead 300–500 μm in size can pass through a 1.8-Fr ultraselective microcatheter in ex vivo study.MethodsEpirubicin (25 mg/1 mL) loaded 100–300 and 300–500 μm DC Bead were tested. Both sizes were diluted 5, 10, and 30 times using contrast material. Ultraselective microcatheter with the outer diameter of 1.8 Fr and the inner diameter of .017 inch (431.8 μm) was used. The diluted DC Bead was injected at a speed of 1 mL/min, and the pressure was continuously measured. The microspheres’ shapes after ejection were observed by a stereomicroscope.ResultsThe maximum pressure of contrast material alone was 8.40 ± 0.21 psi.more » The maximum pressure in 5, 10, and 30 times dilution groups of 100–300 μm were 9.67 ± 1.18, 9.25 ± 0.25, and 9.71 ± 0.28 psi, respectively, whereas 21.10 ± 10.2, 10.48 ± 2.14, 10.09 ± 0.37 psi, respectively in 300–500 μm groups. The maximum pressure in 5 times dilution group of 300–500 μm was significantly higher than the other groups (P < 0.05). In 300–500 μm, 4 of 10 measurements showed high pressure over 24 psi (the maximum value was 43.5 psi) in 5 times dilution group, whereas in 10 times and 30 times dilution groups, all measurements showed less than 12 psi. No damages of microspheres were found.ConclusionsEpirubicin loaded DC Bead 300–500 μm in size can pass through a 1.8-Fr ultraselective microcatheter. To avoid high resistance due to microspheres’ aggregation, dilution more than 10 times is needed.« less

Quantification of Nitrous Oxide from Fugitive Emissions by Tracer Dilution Method using a Mobile Real-time Nitrous Oxide Analyzer

NASA Astrophysics Data System (ADS)

Mønster, J.; Rella, C.; Jacobson, G. A.; He, Y.; Hoffnagle, J.; Scheutz, C.

2012-12-01

Nitrous oxide is a powerful greenhouse gas considered 298 times stronger than carbon dioxide on a hundred years term (Solomon et al. 2007). The increasing global concentration is of great concern and is receiving increasing attention in various scientific and industrial fields. Nitrous oxide is emitted from both natural and anthropogenic sources. Inventories of source specific fugitive nitrous oxide emissions are often estimated on the basis of modeling and mass balance. While these methods are well-developed, actual measurements for quantification of the emissions can be a useful tool for verifying the existing estimation methods as well as providing validation for initiatives targeted at lowering unwanted nitrous oxide emissions. One approach to performing such measurements is the tracer dilution method (Galle et al. 2001), in which a tracer gas is released at the source location at a known flow. The ratio of downwind concentrations of both the tracer gas and nitrous oxide gives the ratios of the emissions rates. This tracer dilution method can be done with both stationary and mobile measurements; in either case, real-time measurements of both tracer and analyte gas is required, which places high demands on the analytical detection method. To perform the nitrous oxide measurements, a novel, robust instrument capable of real-time nitrous oxide measurements has been developed, based on cavity ring-down spectroscopy and operating in the near-infrared spectral region. We present the results of the laboratory and field tests of this instrument in both California and Denmark. Furthermore, results are presented from measurements using the mobile plume method with a tracer gas (acetylene) to quantify the nitrous oxide and methane emissions from known sources such as waste water treatment plants and composting facilities. Nitrous oxide (blue) and methane (yellow) plumes downwind from a waste water treatment facility.

Development and application of molecularly imprinted polymer - Mn-doped ZnS quantum dot fluorescent optosensing for cocaine screening in oral fluid and serum.

PubMed

Chantada-Vázquez, María Pilar; de-Becerra-Sánchez, Carolina; Fernández-Del-Río, Alba; Sánchez-González, Juan; Bermejo, Ana María; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

2018-05-01

A molecularly imprinted polymer - Mn-doped ZnS quantum dot-based fluorescence probe for cocaine abuse screening has been prepared and applied to complex samples such as serum and oral fluid. The fluorescent sensing material was prepared by anchoring a selective MIP for COC on the surface of polyethylene glycol (PEG) modified Mn-doped ZnS quantum dots (QDs). Simple and low cost methods have thus been optimized for assessing cocaine abuse in serum and oral fluid by monitoring fluorescence quenching when cocaine (COC) is present (optimized operating conditions with 1.5mL of 200mgL -1 MIP-coated QDs solution, pH 5.5, and 15min before fluorescence scanning). The matrix effect was found to be important when analyzing oral fluid and serum, and several strategies based on centrifugation for oral fluid and solid phase extraction (SPE) for serum were explored. Two analytical methods were developed for oral fluid. The first one (direct method) requires a centrifugation step (6°C, 4000rpm, 20min) to avoid the matrix effect, and allows for cocaine determination by using an aqueous calibration (1:20 dilution). The second method was developed for oral fluid sampled by Salivette devices, and also requires a further centrifugation (6°C, 4000rpm, 20min) of the recovered oral fluid. This method, however, requires the standard addition technique (1:20 dilution) because of the existence of the matrix effect. Regarding serum samples, a direct method (serum dilution) was not possible, and an SPE procedure was needed to avoid the matrix effect (use of aqueous calibration). The limits of detection and quantification when using the Salivette method were 0.035mgL -1 and 0.117mgL -1, respectively; whereas, 0.015mgL -1 (LOD) and 0.050mgL -1 (LOQ) were obtained for serum. Copyright © 2018 Elsevier B.V. All rights reserved.

The Effect of Dilution on the Structure of Microbial Communities

NASA Technical Reports Server (NTRS)

Mills, Aaron L.

2000-01-01

To determine how dilution of microbial communities affects the diversity of the diluted assemblage a series of numerical simulations were conducted that determined the theoretical change in diversity, richness, and evenness of the community with serial dilution. The results of the simulation suggested that the effects are non linear with a high degree of dependence on the initial evenness of the community being diluted. A series of incubation experiments using a range of dilutions of raw sewage as an inoculum into sterile sewage was used for comparison to the simulations. The diluted communities were maintained in batch fed reactors (three day retention time) for nine days. The communities were harvested and examined by conventional plating and by molecular analysis of the whole-community DNA using AFLP and T-RFLP. Additional, CLPP analysis was also applied. The effects on richness predicted by the numerical simulations were confirmed by the analyses used. The diluted communities fell into three groups, a low dilution, intermediate dilution, and high dilution group, which corresponded well with the groupings obtained for community richness in simulation. The grouping demonstrated the non-linear nature of dilution of whole communities. Furthermore, the results implied that the undiluted community consisted of a few dominant types accompanied by a number of rare (low abundance) types as is typical in unevenly distributed communities.

21 CFR 177.1960 - Vinyl chloride-hexene-1 copolymers.

Code of Federal Regulations, 2014 CFR

2014-04-01

... determined by any suitable analytical procedure of generally accepted applicability. (ii) Inherent viscosity... D1243-79, “Standard Test Method for Dilute Solution Viscosity of Vinyl Chloride Polymers,” which is...

Regeneration of strong-base anion-exchange resins by sequential chemical displacement

DOEpatents

Brown, Gilbert M.; Gu, Baohua; Moyer, Bruce A.; Bonnesen, Peter V.

2002-01-01

A method for regenerating strong-base anion exchange resins utilizing a sequential chemical displacement technique with new regenerant formulation. The new first regenerant solution is composed of a mixture of ferric chloride, a water-miscible organic solvent, hydrochloric acid, and water in which tetrachloroferrate anion is formed and used to displace the target anions on the resin. The second regenerant is composed of a dilute hydrochloric acid and is used to decompose tetrachloroferrate and elute ferric ions, thereby regenerating the resin. Alternative chemical displacement methods include: (1) displacement of target anions with fluoroborate followed by nitrate or salicylate and (2) displacement of target anions with salicylate followed by dilute hydrochloric acid. The methodology offers an improved regeneration efficiency, recovery, and waste minimization over the conventional displacement technique using sodium chloride (or a brine) or alkali metal hydroxide.

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

Hayden, Patrick; Winter, Andreas; Department of Computer Science, University of Bristol, Merchant Venturers Building, Woodland Road, Bristol BS8 1UB

We study the amount of communication needed for two parties to transform some given joint pure state into another one, either exactly or with some fidelity. Specifically, we present a method to lower bound this communication cost even when the amount of entanglement does not increase. Moreover, the bound applies even if the initial state is supplemented with unlimited entanglement in the form of EPR (Einstein-Podolsky-Rosen) pairs and the communication is allowed to be quantum mechanical. We then apply the method to the determination of the communication cost of asymptotic entanglement concentration and dilution. While concentration is known to requiremore » no communication whatsoever, the best known protocol for dilution, discovered by H.-K. Lo and S. Popescu [Phys. Rev. Lett. 83, 1459 (1999)], requires exchange of a number of bits that is of the order of the square root of the number of EPR pairs. Here we prove a matching lower bound of the same asymptotic order, demonstrating the optimality of the Lo-Popescu protocol up to a constant factor and establishing the existence of a fundamental asymmetry between the concentration and dilution tasks. We also discuss states for which the minimal communication cost is proportional to their entanglement, such as the states recently introduced in the context of 'embezzling entanglement' (W. van Dam and P. Hayden, e-print quant-ph/0201041)« less

Highly sensitive and selective hyphenated technique (molecularly imprinted polymer solid-phase microextraction-molecularly imprinted polymer sensor) for ultra trace analysis of aspartic acid enantiomers.

PubMed

Prasad, Bhim Bali; Srivastava, Amrita; Tiwari, Mahavir Prasad

2013-03-29

The present work is related to combination of molecularly imprinted solid-phase microextraction and complementary molecularly imprinted polymer-sensor. The molecularly imprinted polymer grafted on titanium dioxide modified silica fiber was used for microextraction, while the same polymer immobilized on multiwalled carbon nanotubes/titanium dioxide modified pencil graphite electrode served as a detection tool. In both cases, the surface initiated polymerization was found to be advantageous to obtain a nanometer thin imprinted film. The modified silica fiber exhibited high adsorption capacity and enantioselective diffusion of aspartic acid isomers into respective molecular cavities. This combination enabled double preconcentrations of d- and l-aspartic acid that helped sensing both isomers in real samples, without any cross-selectivity and matrix complications. Taking into account 6×10(4)-fold dilution of serum and 2×10(3)-fold dilution of cerebrospinal fluid required by the proposed method, the limit of detection for l-aspartic acid is 0.031ngmL(-1). Also, taking into account 50-fold dilution required by the proposed method, the limit of detection for d-aspartic acid is 0.031ngmL(-1) in cerebrospinal fluid. Copyright © 2013 Elsevier B.V. All rights reserved.

Immune networks: multitasking capabilities near saturation

NASA Astrophysics Data System (ADS)

Agliari, E.; Annibale, A.; Barra, A.; Coolen, A. C. C.; Tantari, D.

2013-10-01

Pattern-diluted associative networks were recently introduced as models for the immune system, with nodes representing T-lymphocytes and stored patterns representing signalling protocols between T- and B-lymphocytes. It was shown earlier that in the regime of extreme pattern dilution, a system with NT T-lymphocytes can manage a number N_B={ {O}}(N_T^\\delta ) of B-lymphocytes simultaneously, with δ < 1. Here we study this model in the extensive load regime NB = αNT, with a high degree of pattern dilution, in agreement with immunological findings. We use graph theory and statistical mechanical analysis based on replica methods to show that in the finite-connectivity regime, where each T-lymphocyte interacts with a finite number of B-lymphocytes as NT → ∞, the T-lymphocytes can coordinate effective immune responses to an extensive number of distinct antigen invasions in parallel. As α increases, the system eventually undergoes a second order transition to a phase with clonal cross-talk interference, where the system’s performance degrades gracefully. Mathematically, the model is equivalent to a spin system on a finitely connected graph with many short loops, so one would expect the available analytical methods, which all assume locally tree-like graphs, to fail. Yet it turns out to be solvable. Our results are supported by numerical simulations.

The non-contact detection and identification of blood stained fingerprints using visible wavelength reflectance hyperspectral imaging: Part 1.

PubMed

Cadd, Samuel; Li, Bo; Beveridge, Peter; O'Hare, William T; Campbell, Andrew; Islam, Meez

2016-05-01

Blood is one of the most commonly encountered types of biological evidence found at scenes of violent crime and one of the most commonly observed fingerprint contaminants. Current visualisation methods rely on presumptive tests or chemical enhancement methods. Although these can successfully visualise ridge detail, they are destructive, do not confirm the presence of blood and can have a negative impact on DNA sampling. A novel application of visible wavelength reflectance hyperspectral imaging (HSI) has been used for the detection and positive identification of blood stained fingerprints in a non-contact and non-destructive manner on white ceramic tiles. The identification of blood was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. HSI has been used to successfully visualise ridge detail in blood stained fingerprints to the ninth depletion. Ridge detail was still detectable with diluted blood to 20-fold dilutions. Latent blood stains were detectable to 15,000-fold dilutions. Ridge detail was detectable for fingerprints up to 6 months old. HSI was also able to conclusively distinguish blood stained fingerprints from fingerprints in six paints and eleven other red/brown media with zero false positives. Copyright © 2016 The Chartered Society of Forensic Sciences. Published by Elsevier Ireland Ltd. All rights reserved.

Comprehensive Study of Volatile Compounds in Two Australian Rosé Wines: Aroma Extract Dilution Analysis (AEDA) of Extracts Prepared Using Solvent-Assisted Flavor Evaporation (SAFE) or Headspace Solid-Phase Extraction (HS-SPE).

PubMed

Wang, Jiaming; Gambetta, Joanna M; Jeffery, David W

2016-05-18

Two rosé wines, representing a tropical and a fruity/floral style, were chosen from a previous study for further exploration by aroma extract dilution analysis (AEDA) and quantitative analysis. Volatiles were extracted using either liquid-liquid extraction (LLE) followed by solvent-assisted flavor evaporation (SAFE) or a recently developed dynamic headspace (HS) sampling method utilizing solid-phase extraction (SPE) cartridges. AEDA was conducted using gas chromatography-mass spectrometry/olfactometry (GC-MS/O) and a total of 51 aroma compounds with a flavor dilution (FD) factor ≥3 were detected. Quantitative analysis of 92 volatiles was undertaken in both wines for calculation of odor activity values. The fruity and floral wine style was mostly driven by 2-phenylethanol, β-damascenone, and a range of esters, whereas 3-SHA and several volatile acids were seen as essential for the tropical style. When extraction methods were compared, HS-SPE was as efficient as SAFE for extracting most esters and higher alcohols, which were associated with fruity and floral characters, but it was difficult to capture volatiles with greater polarity or higher boiling point that may still be important to perceived wine aroma.

Revision of the NIST Standard for (223)Ra: New Measurements and Review of 2008 Data.

PubMed

Zimmerman, B E; Bergeron, D E; Cessna, J T; Fitzgerald, R; Pibida, L

2015-01-01

After discovering a discrepancy in the transfer standard currently being disseminated by the National Institute of Standards and Technology (NIST), we have performed a new primary standardization of the alpha-emitter (223)Ra using Live-timed Anticoincidence Counting (LTAC) and the Triple-to-Double Coincidence Ratio Method (TDCR). Additional confirmatory measurements were made with the CIEMAT-NIST efficiency tracing method (CNET) of liquid scintillation counting, integral γ-ray counting using a NaI(Tl) well counter, and several High Purity Germanium (HPGe) detectors in an attempt to understand the origin of the discrepancy and to provide a correction. The results indicate that a -9.5 % difference exists between activity values obtained using the former transfer standard relative to the new primary standardization. During one of the experiments, a 2 % difference in activity was observed between dilutions of the (223)Ra master solution prepared using the composition used in the original standardization and those prepared using 1 mol·L(-1) HCl. This effect appeared to be dependent on the number of dilutions or the total dilution factor to the master solution, but the magnitude was not reproducible. A new calibration factor ("K-value") has been determined for the NIST Secondary Standard Ionization Chamber (IC "A"), thereby correcting the discrepancy between the primary and secondary standards.

New strategy for determination of anthocyanins, polyphenols and antioxidant capacity of Brassica oleracea liquid extract using infrared spectroscopies and multivariate regression

NASA Astrophysics Data System (ADS)

de Oliveira, Isadora R. N.; Roque, Jussara V.; Maia, Mariza P.; Stringheta, Paulo C.; Teófilo, Reinaldo F.

2018-04-01

A new method was developed to determine the antioxidant properties of red cabbage extract (Brassica oleracea) by mid (MID) and near (NIR) infrared spectroscopies and partial least squares (PLS) regression. A 70% (v/v) ethanolic extract of red cabbage was concentrated to 9° Brix and further diluted (12 to 100%) in water. The dilutions were used as external standards for the building of PLS models. For the first time, this strategy was applied for building multivariate regression models. Reference analyses and spectral data were obtained from diluted extracts. The determinate properties were total and monomeric anthocyanins, total polyphenols and antioxidant capacity by ABTS (2,2-azino-bis(3-ethyl-benzothiazoline-6-sulfonate)) and DPPH (2,2-diphenyl-1-picrylhydrazyl) methods. Ordered predictors selection (OPS) and genetic algorithm (GA) were used for feature selection before PLS regression (PLS-1). In addition, a PLS-2 regression was applied to all properties simultaneously. PLS-1 models provided more predictive models than did PLS-2 regression. PLS-OPS and PLS-GA models presented excellent prediction results with a correlation coefficient higher than 0.98. However, the best models were obtained using PLS and variable selection with the OPS algorithm and the models based on NIR spectra were considered more predictive for all properties. Then, these models provided a simple, rapid and accurate method for determination of red cabbage extract antioxidant properties and its suitability for use in the food industry.

Botulinum Neurotoxins: Qualitative and Quantitative Analysis Using the Mouse Phrenic Nerve Hemidiaphragm Assay (MPN).

PubMed

Bigalke, Hans; Rummel, Andreas

2015-11-25

The historical method for the detection of botulinum neurotoxin (BoNT) is represented by the mouse bioassay (MBA) measuring the animal survival rate. Since the endpoint of the MBA is the death of the mice due to paralysis of the respiratory muscle, an ex vivo animal replacement method, called mouse phrenic nerve (MPN) assay, employs the isolated N. phrenicus-hemidiaphragm tissue. Here, BoNT causes a dose-dependent characteristic decrease of the contraction amplitude of the indirectly stimulated muscle. Within the EQuATox BoNT proficiency 13 test samples were analysed using the MPN assay by serial dilution to a bath concentration resulting in a paralysis time within the range of calibration curves generated with BoNT/A, B and E standards, respectively. For serotype identification the diluted samples were pre-incubated with polyclonal anti-BoNT/A, B or E antitoxin or a combination of each. All 13 samples were qualitatively correctly identified thereby delivering superior results compared to single in vitro methods like LFA, ELISA and LC-MS/MS. Having characterized the BoNT serotype, the final bath concentrations were calculated using the calibration curves and then multiplied by the respective dilution factor to obtain the sample concentration. Depending on the source of the BoNT standards used, the quantitation of ten BoNT/A containing samples delivered a mean z-score of 7 and of three BoNT/B or BoNT/E containing samples z-scores <2, respectively.

Radiotracer Dilution Method for Mercury Inventory Study in Electrolytic Cells

NASA Astrophysics Data System (ADS)

Sugiharto, Su'ud, Zaki; Kurniadi, Rizal; Waris, Abdul; Santoso, Sigit Budi; Abidin, Zainal; Santoso, Gatot Budi

2010-06-01

Purpose of the experiment is to demonstrate feasibility the use of radiotracer to measure weight of mercury in electrolytic cells of soda industry. The weight of mercury in each cell of the plant is designed approximately 1700 kg. Radiotracer is prepared by mixing 203 Hg radioactive mercury with 2400 g of inactive mercury in a bath. The respective precisely weighted mercury aliquots to be injected into the cells are prepared by pouring approximately 130 g of radioactive mercury taken from the bath into 13 standard vials, in accordance with the number of the cells tested. Four standard references prepared by further dilution of ±2 g active mercury taken from the bath to obtain the dilution factors range of 12,000 to 20,000 from which the calibration graph is constructed. The injection process is conducting by pouring the radioactive mercury from aliquots into the flowing mercury at the inlet side of the cell and allows them to mix thoroughly. It is assumed that the mass of the radiotracer injected into a closed system remains constant, at least during the period of the test. From this experiment it was observed that the mixing time is two days after injection of radioactive mercury. The inactive mercury in each electrolytic cell calculated by the radiotracer method is of the range 1351.529 kg to 1966.354 kg with maximum error (95% confidence) is 1.52 %. The accuracy of measurement of the present method is better than gravimetric one which accounts 4 % of error on average.

Determination of cadmium in sediments by diluted HCI extraction and isotope dilution ICP-MS.

PubMed

Terán-Baamonde, Javier; Soto-Ferreiro, Rosa-María; Carlosena, Alatzne; Andrade, José-Manuel; Prada, Darío

2018-08-15

Isotope dilution ICP-MS is proposed to measure the mass fraction of Cd extracted by diluted HCl in marine sediments, using a fast and simple extraction procedure based on ultrasonic probe agitation. The 111 Cd isotope was added before the extraction to achieve isotope equilibration with native Cd solubilized from the sample. The parameters affecting trueness and precision of isotope ratio measurements were evaluated carefully and subsequently corrected in order to minimize errors; they were: detector dead time, spectral interferences, mass discrimination factor and optimum sample/spike ratio. The mass fraction of Cd extracted was compared with the sum of the certified contents of the three steps of the sequential extraction procedure of the Standards, Measurements and Testing Programme (SM&T) analysing the BCR 701 sediment to validate the method. The certified and measured values agreed, giving a measured / certified mass fraction ratio of 1.05. Further, the extraction procedure itself was studied by adding the enriched isotope after the extraction step, which allowed verifying that analyte losses occurred during this process. Two additional reference sediments with certified total cadmium contents were also analysed. The method provided very good precision (0.9%, RSD) and a low detection limit, 1.8 ng g -1 . The procedural uncertainty budget was estimated following the EURACHEM Guide by means of the 'GUM Workbench' software, obtaining a relative expanded uncertainty of 1.5%. The procedure was applied to determine the bioaccessible mass fraction of Cd in sediments from two environmentally and economically important areas of Galicia (rias of Arousa and Vigo, NW of Spain). Copyright © 2018 Elsevier B.V. All rights reserved.

Cell division in Escherichia coli cultures monitored at single cell resolution

PubMed Central

Roostalu, Johanna; Jõers, Arvi; Luidalepp, Hannes; Kaldalu, Niilo; Tenson, Tanel

2008-01-01

Background A fundamental characteristic of cells is the ability to divide. To date, most parameters of bacterial cultures, including cell division, have been measured as cell population averages, assuming that all bacteria divide at a uniform rate. Results We monitored the division of individual cells in Escherichia coli cultures during different growth phases. Our experiments are based on the dilution of green fluorescent protein (GFP) upon cell division, monitored by flow cytometry. The results show that the vast majority of E. coli cells in exponentially growing cultures divided uniformly. In cultures that had been in stationary phase up to four days, no cell division was observed. However, upon dilution of stationary phase culture into fresh medium, two subpopulations of cells emerged: one that started dividing and another that did not. These populations were detectable by GFP dilution and displayed different side scatter parameters in flow cytometry. Further analysis showed that bacteria in the non-growing subpopulation were not dead, neither was the difference in growth capacity reducible to differences in stationary phase-specific gene expression since we observed uniform expression of several stress-related promoters. The presence of non-growing persisters, temporarily dormant bacteria that are tolerant to antibiotics, has previously been described within growing bacterial populations. Using the GFP dilution method combined with cell sorting, we showed that ampicillin lyses growing bacteria while non-growing bacteria retain viability and that some of them restart growth after the ampicillin is removed. Thus, our method enables persisters to be monitored even in liquid cultures of wild type strains in which persister formation has low frequency. Conclusion In principle, the approaches developed here could be used to detect differences in cell division in response to different environmental conditions and in cultures of unicellular organisms other than E. coli. PMID:18430255

A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores

PubMed Central

Nerandzic, Michelle M.; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J.

2016-01-01

Background. Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5–2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods. We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results. Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200–2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions. Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin. PMID:26885539

Quantification of Viral and Prokaryotic Production Rates in Benthic Ecosystems: A Methods Comparison

PubMed Central

Rastelli, Eugenio; Dell’Anno, Antonio; Corinaldesi, Cinzia; Middelboe, Mathias; Noble, Rachel T.; Danovaro, Roberto

2016-01-01

Viruses profoundly influence benthic marine ecosystems by infecting and subsequently killing their prokaryotic hosts, thereby impacting the cycling of carbon and nutrients. Previously conducted studies, based on different methodologies, have provided widely differing estimates of the relevance of viruses on benthic prokaryotes. There has been no attempt so far to compare these independent approaches, including contextual comparisons among different approaches for sample manipulation (i.e., dilution or not of the sediments during incubations), between methods based on epifluorescence microscopy (EFM) or radiotracers, and between the use of different radiotracers. Therefore, it has been difficult to identify the most suitable methodologies and protocols to be used as standard approaches for the quantification of viral infections of prokaryotes. Here, we compared for the first time different methods for determining viral and prokaryotic production rates in marine sediments collected at two benthic sites, differing in depth and environmental conditions. We used a highly replicated experimental design, testing the potential biases associated to the incubation of sediments as diluted or undiluted. In parallel, we also compared EFM counts with the 3H-thymidine incubations for the determination of viral production rates, and the use of 3H-thymidine versus 3H-leucine radiotracers for the determination of prokaryotic production. We show here that, independent from sediment dilution, EFM-based values of viral production ranged from 1.4 to 4.6 × 107 viruses g-1 h-1, and were similar but overall less variable compared to those obtained by the 3H-thymidine method (0.3 to 9.0 × 107 viruses g-1h-1). In addition, the prokaryotic production rates were not affected by sediment dilution, and the use of different radiotracers provided very consistent estimates (10.3–35.1 and 9.3–34.6 ngC g-1h-1 using the 3H-thymidine or 3H-leucine method, respectively). These results indicated that viral lysis was responsible for the abatement of 55–81% of the prokaryotic heterotrophic production, corroborating previous findings of the major role of viruses in benthic deep-sea ecosystems. Moreover, our methodological comparison for the analysis of viral production in marine sediments suggests that microscopy-based approaches are simpler and more cost-effective than those based on radiotracers. These approaches also reduce time to results and overcome issues related to generation of radioactive waste. PMID:27713739

Antimicrobial susceptibility of porcine Brachyspira hyodysenteriae and Brachyspira pilosicoli isolated in Sweden between 1990 and 2010

PubMed Central

2012-01-01

Background The anaerobic spirochetes Brachyspira hyodysenteriae and Brachyspira pilosicoli cause diarrheal diseases in pigs. Their fastidious nature has hampered standardization of methods for antimicrobial susceptibility testing. For monitoring of antimicrobial susceptibility wild type cutoff values are needed to define where the wild type distribution of MICs ends and no approved cutoffs are available for Brachyspira spp. In this study antimicrobial susceptibility data for both species (in total 906 isolates) were compiled and analyzed and wild type cut off values for B. hyodysenteriae proposed. Methods The MICs of tiamulin, valnemulin, tylosin, tylvalosin, doxycycline and lincomycin were determined by broth dilution in brain heart infusion broth supplemented with 10% fetal calf serum. Results The compiled MICs from the broth dilution tests of the B. hyodysenteriae type strain, B78T (ATCC® 27164T), showed that the method yields reproducible results. In an international perspective the frequencies of isolates with decreased antimicrobial susceptibility were low among both B. hyodysenteriae and B. pilosicoli. However, in B. pilosicoli a constant level of 10-15% isolates with tiamulin MICs >4 μg/ml was detected between 2002 and 2010 and in B. hyodysenteriae a gradual increase in tiamulin MICs was seen between 1990 and 2003 although this increase has ceased during the last years. The wild type cutoff values proposed for B. hyodysenteriae are: tiamulin >0.25 μg/ml, valnemulin >0.125 μg/ml, tylosin >16 μg/ml, tylvalosin >1 μg/ml, lincomycin >1 μg/ml and doxycycline >0.5 μg/ml. Conclusions The broth dilution method used in this study has over the years generated tightly grouped MIC populations for the field isolates and reproducible results for the control strain B78T and is therefore a suitable antimicrobial susceptibility test method for monitoring of Brachyspira spp. Here we propose wild type cutoff values for six antimicrobial agents for B. hyodysenteriae tested by broth dilution based on MIC distributions and the current knowledge on mechanisms of resistance in this species. There are few studies on antimicrobial resistance mechanisms and MIC distributions in B. pilosicoli but to some extent the cutoff values proposed for B. hyodysenteriae may be applicable also for monitoring of antimicrobial susceptibility in B. pilosicoli. PMID:22998753

Evaluation of Derivative Ultraviolet Spectrometry for Determining Saccharin in Cola and Other Matrices: An Instrumental Methods Experiment.

ERIC Educational Resources Information Center

Stolzberg, Richard J.

1986-01-01

Background information and experimental procedures are provided for an experiment in which three samples of saccharin (a nickel plating solution, a dilute cola drink, and a more concentrated cola drink) are analyzed and the data interpreted using five methods. Precision and accuracy are evaluated and the best method is selected. (JN)

Online Measurement of Exhaled NO Concentration and Its Production Sites by Fast Non-equilibrium Dilution Ion Mobility Spectrometry

NASA Astrophysics Data System (ADS)

Peng, Liying; Jiang, Dandan; Wang, Zhenxin; Liu, Jiwei; Li, Haiyang

2016-03-01

Exhaled nitric oxide (NO) is one of the most promising breath markers for respiratory diseases. Its profile for exhalation and the respiratory NO production sites can provide useful information for medical disease diagnosis and therapeutic procedures. However, the high-level moisture in exhaled gas always leads to the poor selectivity and sensitivity for ion spectrometric techniques. Herein, a method based on fast non-equilibrium dilution ion mobility spectrometry (NED-IMS) was firstly proposed to directly monitor the exhaled NO profile on line. The moisture interference was eliminated by turbulently diluting the original moisture to 21% of the original with the drift gas and dilution gas. Weak enhancement was observed for humid NO response and its limit of detection at 100% relative humidity was down to 0.58 ppb. The NO concentrations at multiple exhalation flow rates were measured, while its respiratory production sites were determined by using two-compartment model (2CM) and Högman and Meriläinen algorithm (HMA). Last but not the least, the NO production sites were analyzed hourly to tentatively investigate the daily physiological process of NO. The results demonstrated the capacity of NED-IMS in the real-time analysis of exhaled NO and its production sites for clinical diagnosis and assessment.

Normalization of urinary drug concentrations with specific gravity and creatinine.

PubMed

Cone, Edward J; Caplan, Yale H; Moser, Frank; Robert, Tim; Shelby, Melinda K; Black, David L

2009-01-01

Excessive fluid intake can substantially dilute urinary drug concentrations and result in false-negative reports for drug users. Methods for correction ("normalization") of drug/metabolite concentrations in urine have been utilized by anti-doping laboratories, pain monitoring programs, and in environmental monitoring programs to compensate for excessive hydration, but such procedures have not been used routinely in workplace, legal, and treatment settings. We evaluated two drug normalization procedures based on specific gravity and creatinine. These corrections were applied to urine specimens collected from three distinct groups (pain patients, heroin users, and marijuana/ cocaine users). Each group was unique in characteristics, study design, and dosing conditions. The results of the two normalization procedures were highly correlated (r=0.94; range, 0.78-0.99). Increases in percent positives by specific gravity and creatinine normalization were small (0.3% and -1.0%, respectively) for heroin users (normally hydrated subjects), modest (4.2-9.8%) for pain patients (unknown hydration state), and substantial (2- to 38-fold increases) for marijuana/cocaine users (excessively hydrated subjects). Despite some limitations, these normalization procedures provide alternative means of dealing with highly dilute, dilute, and concentrated urine specimens. Drug/metabolite concentration normalization by these procedures is recommended for urine testing programs, especially as a means of coping with dilute specimens.

THE PREPARATION AND PROPERTIES OF HIGHLY PURIFIED ASCORBIC ACID OXIDASE

PubMed Central

Powers, Wendell H.; Lewis, Stanley; Dawson, Charles R.

1944-01-01

1. A method is described for the preparation of a highly purified ascorbic acid oxidase containing 0.24 per cent copper. 2. Using comparable activity measurements, this oxidase is about one and a half times as active on a dry weight basis as the hitherto most highly purified preparation described by Lovett-Janison and Nelson. The latter contained 0.15 per cent copper. 3. The oxidase activity is proportional to the copper content and the proportionality factor is the same as that reported by Lovett-Janison and Nelson. 4. When dialyzed free of salt, the blue concentrated oxidase solutions precipitate a dark green-blue protein which carries the activity. This may be prevented by keeping the concentrated solutions about 0.1 M in Na2HPO4. 5. When highly diluted for activity measurements the oxidase rapidly loses activity (irreversibly) previous to the measurement, unless the dilution is made with a dilute inert protein (gelatin) solution. Therefore activity values obtained using such gelatin-stabilized dilute solutions of the oxidase run considerably higher than values obtained by the Lovett-Janison and Nelson technique. 6. The effect of pH and substrate concentration on the activity of the purified oxidase in the presence and absence of inert protein was studied. PMID:19873382

Simplified spectraphotometric method for the detection of red blood cell agglutination.

PubMed

Ramasubramanian, Melur; Anthony, Steven; Lambert, Jeremy

2008-08-01

Human error is the most significant factor attributed to incompatible blood transfusions. A spectrophotometric approach to blood typing has been developed by examining the spectral slopes of dilute red blood cell (RBC) suspensions in saline, in the presence and absence of various antibodies, offering a technique for the quantitative determination of agglutination intensity [Transfusion39, 1051, 1999TRANAT0041-113210.1046/j.1537-2995.1999.39101051.x]. We offer direct theoretical prediction of the observed change in slope in the 660-1000 nm range through the use of the T-matrix approach and Lorenz-Mie theory for light scattering by dilute RBC suspensions. Following a numerical simulation using the T-matrix code, we present a simplified sensing method for detecting agglutination. The sensor design has been prototyped, fully characterized, and evaluated through a complete set of tests with over 60 RBC samples and compared with the full spectrophotometric method. The LED and photodiode pairs are found to successfully reproduce the spectroscopic determination of red blood cell agglutination.

Simultaneous regularization method for the determination of radius distributions from experimental multiangle correlation functions

NASA Astrophysics Data System (ADS)

Buttgereit, R.; Roths, T.; Honerkamp, J.; Aberle, L. B.

2001-10-01

Dynamic light scattering experiments have become a powerful tool in order to investigate the dynamical properties of complex fluids. In many applications in both soft matter research and industry so-called ``real world'' systems are subject of great interest. Here, the dilution of the investigated system often cannot be changed without getting measurement artifacts, so that one often has to deal with highly concentrated and turbid media. The investigation of such systems requires techniques that suppress the influence of multiple scattering, e.g., cross correlation techniques. However, measurements at turbid as well as highly diluted media lead to data with low signal-to-noise ratio, which complicates data analysis and leads to unreliable results. In this article a multiangle regularization method is discussed, which copes with the difficulties arising from such samples and enhances enormously the quality of the estimated solution. In order to demonstrate the efficiency of this multiangle regularization method we applied it to cross correlation functions measured at highly turbid samples.

Practical issues relating to soil column chromatography for sorption parameter determination.

PubMed

Bi, Erping; Schmidt, Torsten C; Haderlein, Stefan B

2010-08-01

Determination of sorption distribution coefficients (K(d)) of organic compounds by a dynamic soil column chromatography (SCC) method was developed and validated. Eurosoil 4, quartz, and alumina were chosen as exemplary packing materials. Heterocyclic aromatic compounds were selected in the validation of SCC. The prerequisites of SCC with regard to column dimension, packing procedure, and sample injection volume are discussed. Reproducible soil column packing was achieved by addition of a pre-column and an HPLC pump for subsequent compression of the packed material. Various methods to determine retention times from breakthrough curves are discussed and the use of the half mass method is recommended. To dilute soil with inert material can prevent column-clogging and help to complete experiments in a reasonable period of time. For the chosen probe compounds, quartz rather than alumina proved a suitable dilution material. Non-equilibrium issue can be overcome by conducting the experiments under different flowrates and/or performing numerical simulation. Copyright 2010 Elsevier Ltd. All rights reserved.

Cross-correlation focus method with an electrostatic sensor array for local particle velocity measurement in dilute gas-solid two-phase flow

NASA Astrophysics Data System (ADS)

Wang, Chao; Zhang, Jingyu; Gao, Wenbin; Ding, Hongbing; Wu, Weiping

2015-11-01

The gas-solid two-phase flow has been widely applied in the power, chemical and metallurgical industries. It is of great significance in the research of gas-solid two-phase flow to measure particle velocity at different locations in the pipeline. Thus, an electrostatic sensor array comprising eight arc-shaped electrodes was designed. The relationship between the cross-correlation (CC) velocity and the distribution of particle velocity, charge density and electrode spatial sensitivity was analysed. Then the CC sensitivity and its calculation method were proposed. According to the distribution of CC sensitivity, it was found that, between different electrode pairs, it had different focus areas. The CC focus method was proposed for particle velocity measurement at different locations and validated by a belt-style electrostatic induction experiment facility. Finally, the particle velocities at different locations with different flow conditions were measured to research the particle velocity distribution in a dilute horizontal pneumatic conveying pipeline.

Semiautomated Method for Microbiological Vitamin Assays

PubMed Central

Berg, T. M.; Behagel, H. A.

1972-01-01

A semiautomated method for microbiological vitamin assays is described, which includes separate automated systems for the preparation of the cultures and for the measurement of turbidity. In the dilution and dosage unit based on the continuous-flow principle, vitamin samples were diluted to two different dose levels at a rate of 40 per hr, mixed with the inoculated test broth, and dispensed into culture tubes. After incubation, racks with culture tubes were placed on the sampler of an automatic turbidimeter. This unit, based on the discrete-sample system, measured the turbidity and printed the extinction values at a rate of 300 per hr. Calculations were computerized and the results, including statistical data, are presented in an easily readable form. The automated method is in routine use for the assays of thiamine, riboflavine, pyridoxine, cyanocobalamin, calcium pantothenate, nicotinic acid, pantothenol, and folic acid. Identical vitamin solutions assayed on different days gave variation coefficients for the various vitamin assays of less than 10%. Images PMID:4553802

The Effects of Micromixing Two Solutions of Two Concentrations in a Two Tier PDMS Micromixer

NASA Astrophysics Data System (ADS)

Sundra, Sargunan; Fhong Soon, Chin; Zainal, Nurfarina; Sek Tee, Kian; Ahmad, Nornabihah; Gan, Siew Hua

2017-08-01

Micromixing technology has drastically advanced in the past few decades. Micromixers are one of the elements in integrated microfluidic systems for chemical, analytical chemistry, pharmaceutical, and biological applications. In this study, two tier micromixer was used to mix and dilute two solutions of similar and different concentration in order to investigate performance of micromixer’s mixing. This paper presents the fabrication of a designed micromixer using polydimethylsiloxane (PDMS) and vinyl tape methods which reduce time, cost and complexity of prototyping. The serpentine structure of the microchannels was designed to enhance both mixing and dilution. Two types of food dyes and distilled water were used to investigate the mixing performance of the micromixer followed by spectrophotometry analysis. It is observed that the single dye solution and distilled water shows better mixing performance compared to the micromixing of two dye solutions which was supported by the diffusion theory. 2.00 ml/min was the optimum flow rate that allow optimum mixing and dilution between two different concentrated liquids.

Is the gas-particle partitioning in alpha-pinene secondary organic aerosol reversible?

NASA Astrophysics Data System (ADS)

Grieshop, Andrew P.; Donahue, Neil M.; Robinson, Allen L.

2007-07-01

This paper discusses the reversibility of gas-particle partitioning in secondary organic aerosol (SOA) formed from α-pinene ozonolysis in a smog chamber. Previously, phase partitioning has been studied quantitatively via SOA production experiments and qualitatively by perturbing temperature and observing particle evaporation. In this work, two methods were used to isothermally dilute the SOA: an external dilution sampler and an in-chamber technique. Dilution caused some evaporation of SOA, but repartitioning took place on a time scale of tens of minutes to hours-consistent with an uptake coefficient on the order of 0.001-0.01. However, given sufficient time, α-pinene SOA repartitions reversibly based on comparisons with data from conventional SOA yield experiments. Further, aerosol mass spectrometer (AMS) data indicate that the composition of SOA varies with partitioning. These results suggest that oligomerization observed in high-concentration laboratory experiments may be a reversible process and underscore the complexity of the kinetics of formation and evaporation of SOA.

Frustrated spin chains in strong magnetic field: Dilute two-component Bose gas regime

NASA Astrophysics Data System (ADS)

Kolezhuk, A. K.; Heidrich-Meisner, F.; Greschner, S.; Vekua, T.

2012-02-01

We study the ground state of frustrated spin-S chains in a strong magnetic field in the immediate vicinity of saturation. In strongly frustrated chains, the magnon dispersion has two degenerate minima at inequivalent momenta ±Q, and just below the saturation field the system can be effectively represented as a dilute one-dimensional lattice gas of two species of bosons that correspond to magnons with momenta around ±Q. We present a theory of effective interactions in such a dilute magnon gas that allows us to make quantitative predictions for arbitrary values of the spin. With the help of this method, we are able to establish the magnetic phase diagram of frustrated chains close to saturation and study phase transitions between several nontrivial states, including a two-component Luttinger liquid, a vector chiral phase, and phases with bound magnons. We study those phase transitions numerically and find a good agreement with our analytical predictions.

The CETAC ADX-500 Autodiluter System: A Study of Dilution Performance with the ELAN 6000 ICP-MS and ELAN Software

USGS Publications Warehouse

May, T.W.; Wiedmeyer, Ray H.

1998-01-01

The CETAC ADX-500 autodiluter system was tested with ELAN?? v 2.1 software and the ELAN 6000 ICP-MS instrument to determine on-line automated dilution performance during analysis of standard solutions containing nine analytes representative of the mass spectral range (mass 9 to mass 238). Two or more dilution schemes were tested for each of 5 test tube designs. Dilution performance was determined by comparison of analyte concentration means of diluted and non-diluted standards. Accurate dilutions resulted with one syringe pump addition of diluent in small diameter round-bottomed (13 mm OD) or conical-tipped (18 mm OD) tubes and one or more syringe pump additions in large diameter (28 mm OD) conical-tipped tubes. Inadequate dilution mixing which produced high analyte concentration means was observed for all dilutions conducted in flat-bottomed tubes, and for dilutions requiring multiple syringe additions of diluent in small diameter round-bottomed and conical tipped tubes. Effective mixing of diluted solutions was found to depend largely upon tube diameter and liquid depth: smaller tube diameters and greater liquid depth resulted in ineffective mixing, whereas greater tube diameter and shallower liquid depth facilitated effective mixing. Two design changes for the autodiluter were suggested that would allow effective mixing to occur using any dilution scheme and tube design.

40 CFR 799.6755 - TSCA partition coefficient (n-octanol/water), shake flask method.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) Qualifying statements. This method applies only to pure, water soluble substances which do not dissociate or... applied. The values presented in table 1 of this section are not necessarily representative of the results... Law applies only at constant temperature, pressure, and pH for dilute solutions. It strictly applies...

Interferometric studies of the refractive indices of some fluorine compounds

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

Malik, Jim Gorden; Rogers, Max T.; Speirs, John L.

1954-08-01

It is the object of this investigation to construct an interferometer of the Rayleigh type and adapt it for two purposes; first, the measurement of the refractive indices of gases by an absolute method and, and second, the measurement of the refractive indices of very dilute solutions by the difference method.

40 CFR 230.73 - Actions affecting the method of dispersion.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Actions To Minimize Adverse Effects § 230.73 Actions affecting the method of dispersion. The effects of a... obstruction to the water current or circulation pattern, and utilizing natural bottom contours to minimize the... patterns to mix, disperse and dilute the discharge; (e) Minimizing water column turbidity by using a...

40 CFR 230.73 - Actions affecting the method of dispersion.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Actions To Minimize Adverse Effects § 230.73 Actions affecting the method of dispersion. The effects of a... obstruction to the water current or circulation pattern, and utilizing natural bottom contours to minimize the... patterns to mix, disperse and dilute the discharge; (e) Minimizing water column turbidity by using a...

40 CFR 230.73 - Actions affecting the method of dispersion.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Actions To Minimize Adverse Effects § 230.73 Actions affecting the method of dispersion. The effects of a... obstruction to the water current or circulation pattern, and utilizing natural bottom contours to minimize the... patterns to mix, disperse and dilute the discharge; (e) Minimizing water column turbidity by using a...

40 CFR 230.73 - Actions affecting the method of dispersion.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Actions To Minimize Adverse Effects § 230.73 Actions affecting the method of dispersion. The effects of a... obstruction to the water current or circulation pattern, and utilizing natural bottom contours to minimize the... patterns to mix, disperse and dilute the discharge; (e) Minimizing water column turbidity by using a...

21 CFR 177.1570 - Poly-1-butene resins and butene/ethylene copolymers.

Code of Federal Regulations, 2013 CFR

2013-04-01

... their characteristic infrared spectra. (ii) Viscosity. Poly-1-butene resins and the butene/ethylene copolymers have an intrinsic viscosity 1.0 to 3.2 as determined by ASTM method D1601-78, “Standard Test Method for Dilute Solution Viscosity of Ethylene Polymers,” which is incorporated by reference. Copies...

21 CFR 177.1570 - Poly-1-butene resins and butene/ethylene copolymers.

Code of Federal Regulations, 2012 CFR

2012-04-01

... their characteristic infrared spectra. (ii) Viscosity. Poly-1-butene resins and the butene/ethylene copolymers have an intrinsic viscosity 1.0 to 3.2 as determined by ASTM method D1601-78, “Standard Test Method for Dilute Solution Viscosity of Ethylene Polymers,” which is incorporated by reference. Copies...

Measurement of the complex permittivity of dry rocks and minerals: application of polythene dilution method and Lichtenecker's mixture formulae

NASA Astrophysics Data System (ADS)

Zheng, Yongchun; Wang, Shijie; Feng, Junming; Ouyang, Ziyuan; Li, Xiongyao

2005-12-01

The complex permittivity of dry rocks and minerals varies over a very wide range, even within a sample there are variation at different temperatures and frequencies. Most rocks and minerals are inhomogeneous materials, therefore, most of the present methods of dielectric measurement designed for artificial homogeneous materials are not suitable for rocks and minerals. The resonant cavity perturbation (RCP) method is a reliable and simple technique to determine the complex permittivity of dielectric materials in the GHz range, and this method is also used extensively. However, the traditional RCP method is sensitive to the measurement of low dielectric constant (ɛ') and low loss factor (ɛ'' or tanδ) materials. The complex permittivity of most dry rocks and minerals exceeds the span vibration of the RCP method, and cannot be measured by the RCP method directly. This paper proposes a new method to measure the complex permittivity of dry rocks and minerals with the RCP method incorporated in the application of polythene (PE) dilution method and Lichtenecker's mixture formulae. Dry rocks and minerals were ground into fine powder. The powder of rocks and minerals was mixed with polythene powder in a definite volume per cent. The mixture was heated and pressed into a thin circular slice. The slice was processed into a small rectangular strip sample, the size of which was fitted to the demands of the RCP method. The complex permittivity of the strip was obtained by the RCP method. The relationship between the dielectric properties of the two-phase mixture and those of each phase in the mixture can be expressed by Lichtenecker's mixture formula. Thus the complex permittivity of dry rocks and minerals can be calculated from the complex permittivity of the mixture in case the complex permittivity of polythene is known. The presented method was verified by measurements of reference materials of various known complex permittivity and other reliable dielectric measurement methods. The results of the experiment showed that this new method is of high accuracy, small sample requirement, and convenient application. Moreover, the complex permittivity of rocks and minerals measured by this method is more reliable than the direct dielectric measurement of rocks or minerals without application of the polythene dilution method and Lichtenecker's mixture formulae.

Sulfuric acid-sulfur heat storage cycle

DOEpatents

Norman, John H.

1983-12-20

A method of storing heat is provided utilizing a chemical cycle which interconverts sulfuric acid and sulfur. The method can be used to levelize the energy obtained from intermittent heat sources, such as solar collectors. Dilute sulfuric acid is concentrated by evaporation of water, and the concentrated sulfuric acid is boiled and decomposed using intense heat from the heat source, forming sulfur dioxide and oxygen. The sulfur dioxide is reacted with water in a disproportionation reaction yielding dilute sulfuric acid, which is recycled, and elemental sulfur. The sulfur has substantial potential chemical energy and represents the storage of a significant portion of the energy obtained from the heat source. The sulfur is burned whenever required to release the stored energy. A particularly advantageous use of the heat storage method is in conjunction with a solar-powered facility which uses the Bunsen reaction in a water-splitting process. The energy storage method is used to levelize the availability of solar energy while some of the sulfur dioxide produced in the heat storage reactions is converted to sulfuric acid in the Bunsen reaction.

Direct analysis of psychoactive tryptamine and harmala alkaloids in the Amazonian botanical medicine ayahuasca by liquid chromatography-electrospray ionization-tandem mass spectrometry.

PubMed

McIlhenny, Ethan H; Pipkin, Kelly E; Standish, Leanna J; Wechkin, Hope A; Strassman, Rick; Barker, Steven A

2009-12-18

A direct injection/liquid chromatography-electrospray ionization-tandem mass spectrometry procedure has been developed for the simultaneous quantitation of 11 compounds potentially found in the increasingly popular Amazonian botanical medicine and religious sacrament ayahuasca. The method utilizes a deuterated internal standard for quantitation and affords rapid detection of the alkaloids by a simple dilution assay, requiring no extraction procedures. Further, the method demonstrates a high degree of specificity for the compounds in question, as well as low limits of detection and quantitation despite using samples for analysis that had been diluted up to 200:1. This approach also appears to eliminate potential matrix effects. Method bias for each compound, examined over a range of concentrations, was also determined as was inter- and intra-assay variation. Its application to the analysis of three different ayahuasca preparations is also described. This method should prove useful in the study of ayahuasca in clinical and ethnobotanical research as well as in forensic examinations of ayahuasca preparations.

Multi-mycotoxin stable isotope dilution LC-MS/MS method for Fusarium toxins in beer.

PubMed

Habler, Katharina; Gotthardt, Marina; Schüler, Jan; Rychlik, Michael

2017-03-01

A stable isotope dilution LC-MS/MS multi-mycotoxin method was developed for 12 different Fusarium toxins including modified mycotoxins in beer (deoxynivalenol-3-glucoside, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyl-deoxynivalenol, HT2-toxin, T2-toxin, enniatin B, B1, A1, A, beauvericin and zearalenone). As sample preparation and purification of beer a combined solid phase extraction for trichothecenes, enniatins, beauvericin and zearalenone was firstly developed. The validation of the new method gave satisfying results: intra-day and inter-day precision and recoveries were 1-5%, 2-8% and 72-117%, respectively. In total, 61 different organic and conventional beer samples from Germany and all over the world were analyzed by using the newly developed multi-mycotoxin method. In summary, deoxynivalenol, deoxynivalenol-3-glucoside, 3-acetyldeoxynivaleneol and enniatin B were quantified in rather low contents in the investigated beer samples. None of the other monitored Fusarium toxins like 15-acetyldeoxynivalenol, HT2- and T2-toxin, zearalenone, enniatin B1, A1, A or beauvericin were detectable. Copyright © 2016 Elsevier Ltd. All rights reserved.

High-performance liquid chromatography with fluorescence detection for the rapid analysis of pheophytins and pyropheophytins in virgin olive oil.

PubMed

Li, Xueqi; Woodman, Michael; Wang, Selina C

2015-08-01

Pheophytins and pyropheophytin are degradation products of chlorophyll pigments, and their ratios can be used as a sensitive indicator of stress during the manufacturing and storage of olive oil. They increase over time depending on the storage condition and if the oil is exposed to heat treatments during the refining process. The traditional analysis method includes solvent- and time-consuming steps of solid-phase extraction followed by analysis by high-performance liquid chromatography with ultraviolet detection. We developed an improved dilute/fluorescence method where multi-step sample preparation was replaced by a simple isopropanol dilution before the high-performance liquid chromatography injection. A quaternary solvent gradient method was used to include a fourth strong solvent wash on a quaternary gradient pump, which avoided the need to premix any solvents and greatly reduced the oil residues on the column from previous analysis. This new method not only reduces analysis cost and time but shows reliability, repeatability, and improved sensitivity, especially important for low-level samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Inorganic arsenic in seafood: does the extraction method matter?

PubMed

Pétursdóttir, Ásta H; Gunnlaugsdóttir, Helga; Krupp, Eva M; Feldmann, Jörg

2014-05-01

Nine different extraction methods were evaluated for three seafood samples to test whether the concentration of inorganic arsenic (iAs) determined in seafood is dependent on the extraction method. Certified reference materials (CRM) DOLT-4 (Dogfish Liver) and TORT-2 (Lobster Hepatopancreas), and a commercial herring fish meal were evaluated. All experimental work described here was carried out by the same operator using the same instrumentation, thus eliminating possible differences in results caused by laboratory related factors. Low concentrations of iAs were found in CRM DOLT-4 (0.012±0.003mgkg(-1)) and the herring fish meal sample (0.007±0.002mgkg(-1)) for all extraction methods. When comparing the concentration of iAs in CRM TORT-2 found in this study and in the literature dilute acids, HNO3 and HCl, showed the highest extracted iAs wheras dilute NaOH (in 50% ethanol) showed significantly lower extracted iAs. However, most other extraction solvents were not statistically different from one another. Copyright © 2013 Elsevier Ltd. All rights reserved.

The effect of dilution and the use of a post-extraction nucleic acid purification column on the accuracy, precision, and inhibition of environmental DNA samples

USGS Publications Warehouse

Mckee, Anna M.; Spear, Stephen F.; Pierson, Todd W.

2015-01-01

Isolation of environmental DNA (eDNA) is an increasingly common method for detecting presence and assessing relative abundance of rare or elusive species in aquatic systems via the isolation of DNA from environmental samples and the amplification of species-specific sequences using quantitative PCR (qPCR). Co-extracted substances that inhibit qPCR can lead to inaccurate results and subsequent misinterpretation about a species’ status in the tested system. We tested three treatments (5-fold and 10-fold dilutions, and spin-column purification) for reducing qPCR inhibition from 21 partially and fully inhibited eDNA samples collected from coastal plain wetlands and mountain headwater streams in the southeastern USA. All treatments reduced the concentration of DNA in the samples. However, column purified samples retained the greatest sensitivity. For stream samples, all three treatments effectively reduced qPCR inhibition. However, for wetland samples, the 5-fold dilution was less effective than other treatments. Quantitative PCR results for column purified samples were more precise than the 5-fold and 10-fold dilutions by 2.2× and 3.7×, respectively. Column purified samples consistently underestimated qPCR-based DNA concentrations by approximately 25%, whereas the directional bias in qPCR-based DNA concentration estimates differed between stream and wetland samples for both dilution treatments. While the directional bias of qPCR-based DNA concentration estimates differed among treatments and locations, the magnitude of inaccuracy did not. Our results suggest that 10-fold dilution and column purification effectively reduce qPCR inhibition in mountain headwater stream and coastal plain wetland eDNA samples, and if applied to all samples in a study, column purification may provide the most accurate relative qPCR-based DNA concentrations estimates while retaining the greatest assay sensitivity.

Thermo-kinetic instabilities in model reactors. Examples in experimental tests

NASA Astrophysics Data System (ADS)

Lavadera, Marco Lubrano; Sorrentino, Giancarlo; Sabia, Pino; de Joannon, Mara; Cavaliere, Antonio; Ragucci, Raffaele

2017-11-01

The use of advanced combustion technologies (such as MILD, LTC, etc.) is among the most promising methods to reduce emission of pollutants. For such technologies, working temperatures are enough low to boost the formation of several classes of pollutants, such as NOx and soot. To access this temperature range, a significant dilution as well as preheating of reactants is required. Such conditions are usually achieved by a strong recirculation of exhaust gases that simultaneously dilute and pre-heat the fresh reactants. These peculiar operative conditions also imply strong fuel flexibility, thus allowing the use of low calorific value (LCV) energy carriers with high efficiency. However, the intersection of low combustion temperatures and highly diluted mixtures with intense pre-heating alters the evolution of the combustion process with respect to traditional flames, leading to features such as the susceptibility to oscillations, which are undesirable during combustion. Therefore, an effective use of advanced combustion technologies requires a thorough analysis of the combustion kinetic characteristics in order to identify optimal operating conditions and control strategies with high efficiency and low pollutant emissions. The present work experimentally and numerically characterized the ignition and oxidation processes of methane and propane, highly diluted in nitrogen, at atmospheric pressure, in a Plug Flow Reactor and a Perfectly Stirred Reactor under a wide range of operating conditions involving temperatures, mixture compositions and dilution levels. The attention was focused particularly on the chemistry of oscillatory phenomena and multistage ignitions. The global behavior of these systems can be qualitatively and partially quantitatively modeled using the detailed kinetic models available in the literature. Results suggested that, for diluted conditions and lower adiabatic flame temperatures, the competition among several pathways, i.e. intermediate- and high-temperature branching, branching and recombination channels, oxidation and recombination/pyrolysis pathways, is enhanced, thus permitting the onset of phenomena that are generally hidden during conventional combustion processes.

Extreme sensitivity of gene expression in human SH-SY5Y neurocytes to ultra-low doses of Gelsemium sempervirens

PubMed Central

2014-01-01

Background Gelsemium sempervirens L. (Gelsemium s.) is a traditional medicinal plant, employed as an anxiolytic at ultra-low doses and animal models recently confirmed this activity. However the mechanisms by which it might operate on the nervous system are largely unknown. This work investigates the gene expression of a human neurocyte cell line treated with increasing dilutions of Gelsemium s. extract. Methods Starting from the crude extract, six 100 × (centesimal, c) dilutions of Gelsemium s. (2c, 3c, 4c, 5c, 9c and 30c) were prepared according to the French homeopathic pharmacopoeia. Human SH-SY5Y neuroblastoma cells were exposed for 24 h to test dilutions, and their transcriptome compared by microarray to that of cells treated with control vehicle solutions. Results Exposure to the Gelsemium s. 2c dilution (the highest dose employed, corresponding to a gelsemine concentration of 6.5 × 10-9 M) significantly changed the expression of 56 genes, of which 49 were down-regulated and 7 were overexpressed. Several of the down-regulated genes belonged to G-protein coupled receptor signaling pathways, calcium homeostasis, inflammatory response and neuropeptide receptors. Fisher exact test, applied to the group of 49 genes down-regulated by Gelsemium s. 2c, showed that the direction of effects was significantly maintained across the treatment with high homeopathic dilutions, even though the size of the differences was distributed in a small range. Conclusions The study shows that Gelsemium s., a medicinal plant used in traditional remedies and homeopathy, modulates a series of genes involved in neuronal function. A small, but statistically significant, response was detected even to very low doses/high dilutions (up to 30c), indicating that the human neurocyte genome is extremely sensitive to this regulation. PMID:24642002

21 CFR 864.5240 - Automated blood cell diluting apparatus.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated blood cell diluting apparatus. 864.5240... § 864.5240 Automated blood cell diluting apparatus. (a) Identification. An automated blood cell diluting apparatus is a fully automated or semi-automated device used to make appropriate dilutions of a blood sample...

21 CFR 864.5240 - Automated blood cell diluting apparatus.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Automated blood cell diluting apparatus. 864.5240... § 864.5240 Automated blood cell diluting apparatus. (a) Identification. An automated blood cell diluting apparatus is a fully automated or semi-automated device used to make appropriate dilutions of a blood sample...

Hybridizing pines with diluted pollen

Treesearch

Robert Z. Callaham

1967-01-01

Diluted pollens would have many uses by the tree breeder. Dilutions would be particularly advantageous in making many controlled pollinations with a limited amount of pollen. They also would be useful in artificial mass pollinations of orchards or single trees. Diluted pollens might help overcome troublesome genetic barriers to crossing. Feasibility o,f using diluted...

Sodium hypochlorite (dilute chlorine bleach) oral rinse in patient self-care.

PubMed

Rich, Sandra K; Slots, Jørgen

2015-01-01

Sodium hypochlorite (NaOCl), commonly known as "bleach," is widely accepted as being a safe and effective antiseptic against bacteria, fungi, and viruses. For over a century, bleach has been used to control or overcome infection in homes, hospitals, and even on battlefields, and in endodontics for disinfection of root canals. This paper reviews clinical studies on the efficacy of sodium hypochlorite oral rinse to combat dental plaque and gingival inflammation. Sodium hypochlorite is readily available as inexpensive household bleach, and we suggest that oral rinsing twice weekly with dilute bleach (0.25% sodium hypochlorite) constitutes a valuable adjunct to current methods of plaque removal.

Ion exchange purification of scandium

DOEpatents

Herchenroeder, Laurie A.; Burkholder, Harvey R.

1990-10-23

An improvement in purification of scandium through ion exchange chromatography is disclosed in which the oxidation potential of the eluting solution is altered by the addition of potassium chlorate or ammonium chloride so that removal of contaminants is encouraged. The temperature, pH and concentration of the eluent HEDTA are controlled in order to maintain the scandium in the column while minimizing dilution of the scandium band. Recovery of scandium is improved by pumping dilute scandium over the column prior to stripping the scandium and precipitation. This eliminates the HEDTA ion and other monovalent cations contaminating the scandium band. This method maximizes recovery of scandium while maintaining purity.

Ion exchange purification of scandium

DOEpatents

Herchenroeder, L.A.; Burkholder, H.R.

1990-10-23

An improvement in purification of scandium through ion exchange chromatography is disclosed in which the oxidation potential of the eluting solution is altered by the addition of potassium chlorate or ammonium chloride so that removal of contaminants is encouraged. The temperature, pH and concentration of the eluent HEDTA are controlled in order to maintain the scandium in the column while minimizing dilution of the scandium band. Recovery of scandium is improved by pumping dilute scandium over the column prior to stripping the scandium and precipitation. This eliminates the HEDTA ion and other monovalent cations contaminating the scandium band. This method maximizes recovery of scandium while maintaining purity. 2 figs.

Phase transition in nonuniform Josephson arrays: Monte Carlo simulations

NASA Astrophysics Data System (ADS)

Lozovik, Yu. E.; Pomirchy, L. M.

1994-01-01

Disordered 2D system with Josephson interactions is considered. Disordered XY-model describes the granular films, Josephson arrays etc. Two types of disorder are analyzed: (1) randomly diluted system: Josephson coupling constants J ij are equal to J with probability p or zero (bond percolation problem); (2) coupling constants J ij are positive and distributed randomly and uniformly in some interval either including the vicinity of zero or apart from it. These systems are simulated by Monte Carlo method. Behaviour of potential energy, specific heat, phase correlation function and helicity modulus are analyzed. The phase diagram of the diluted system in T c-p plane is obtained.

High-throughput ab-initio dilute solute diffusion database.

PubMed

Wu, Henry; Mayeshiba, Tam; Morgan, Dane

2016-07-19

We demonstrate automated generation of diffusion databases from high-throughput density functional theory (DFT) calculations. A total of more than 230 dilute solute diffusion systems in Mg, Al, Cu, Ni, Pd, and Pt host lattices have been determined using multi-frequency diffusion models. We apply a correction method for solute diffusion in alloys using experimental and simulated values of host self-diffusivity. We find good agreement with experimental solute diffusion data, obtaining a weighted activation barrier RMS error of 0.176 eV when excluding magnetic solutes in non-magnetic alloys. The compiled database is the largest collection of consistently calculated ab-initio solute diffusion data in the world.

Diluted magnetic semiconductors with narrow band gaps

NASA Astrophysics Data System (ADS)

Gu, Bo; Maekawa, Sadamichi

2016-10-01

We propose a method to realize diluted magnetic semiconductors (DMSs) with p - and n -type carriers by choosing host semiconductors with a narrow band gap. By employing a combination of the density function theory and quantum Monte Carlo simulation, we demonstrate such semiconductors using Mn-doped BaZn2As2 , which has a band gap of 0.2 eV. In addition, we found a nontoxic DMS Mn-doped BaZn2Sb2 , of which the Curie temperature Tc is predicted to be higher than that of Mn-doped BaZn2As2 , the Tc of which was up to 230 K in a recent experiment.

Constraints on dilution from a narrow attentional zoom reveal how spatial and color cues direct selection.

PubMed

Chen, Zhe; Cave, Kyle R

2014-08-01

Distractor interference is subject to dilution from other nontarget elements, and the level of dilution is affected by attention. This study explores the nature of dilution when the location and color of the target is known in advance. Experiments 1 and 2 show that attention is effectively limited to the precued region, so that it is the nontarget letters appearing at the cued locations that are responsible for most of the dilution, and not those appearing at the uncued locations. Furthermore, this dilution occurs relatively early in processing. Experiment 3 demonstrates that top-down attentional control can prevent dilution, because foreknowledge of the target color leads to quick attention shifts. Experiment 4 illustrates bottom-up attentional control in preventing dilution when the distractor is a color singleton that is segregated from the diluting nontargets. The results show that dilution is modulated by both top-down and bottom-up factors, that it can occur even when attention is restricted to a relatively small region, and that it occurs early in processing, but not so early that it avoids the effects of attention. They provide new challenges for earlier accounts suggesting that dilution is widespread and unfettered by attention. Likewise, some parts of the results are difficult to reconcile with the alternative perceptual load theory, but they do support a form of dilution that is limited by attentional boundaries. Because of that link to attention, dilution is a useful tool for measuring how attention is guided by information about target location and color. Copyright © 2014 Elsevier Ltd. All rights reserved.

Automated PCR setup for forensic casework samples using the Normalization Wizard and PCR Setup robotic methods.

PubMed

Greenspoon, S A; Sykes, K L V; Ban, J D; Pollard, A; Baisden, M; Farr, M; Graham, N; Collins, B L; Green, M M; Christenson, C C

2006-12-20

Human genome, pharmaceutical and research laboratories have long enjoyed the application of robotics to performing repetitive laboratory tasks. However, the utilization of robotics in forensic laboratories for processing casework samples is relatively new and poses particular challenges. Since the quantity and quality (a mixture versus a single source sample, the level of degradation, the presence of PCR inhibitors) of the DNA contained within a casework sample is unknown, particular attention must be paid to procedural susceptibility to contamination, as well as DNA yield, especially as it pertains to samples with little biological material. The Virginia Department of Forensic Science (VDFS) has successfully automated forensic casework DNA extraction utilizing the DNA IQ(trade mark) System in conjunction with the Biomek 2000 Automation Workstation. Human DNA quantitation is also performed in a near complete automated fashion utilizing the AluQuant Human DNA Quantitation System and the Biomek 2000 Automation Workstation. Recently, the PCR setup for casework samples has been automated, employing the Biomek 2000 Automation Workstation and Normalization Wizard, Genetic Identity version, which utilizes the quantitation data, imported into the software, to create a customized automated method for DNA dilution, unique to that plate of DNA samples. The PCR Setup software method, used in conjunction with the Normalization Wizard method and written for the Biomek 2000, functions to mix the diluted DNA samples, transfer the PCR master mix, and transfer the diluted DNA samples to PCR amplification tubes. Once the process is complete, the DNA extracts, still on the deck of the robot in PCR amplification strip tubes, are transferred to pre-labeled 1.5 mL tubes for long-term storage using an automated method. The automation of these steps in the process of forensic DNA casework analysis has been accomplished by performing extensive optimization, validation and testing of the software methods.

Analysis of nitrosamines in water by automated SPE and isotope dilution GC/HRMS Occurrence in the different steps of a drinking water treatment plant, and in chlorinated samples from a reservoir and a sewage treatment plant effluent.

PubMed

Planas, Carles; Palacios, Oscar; Ventura, Francesc; Rivera, Josep; Caixach, Josep

2008-08-15

A method based on automated solid-phase extraction (SPE) and isotope dilution gas chromatography/high resolution mass spectrometry (GC/HRMS) has been developed for the analysis of nine nitrosamines in water samples. The combination of automated SPE and GC/HRMS for the analysis of nitrosamines has not been reported previously. The method shows as advantages the selectivity and sensitivity of GC/HRMS analysis and the high efficiency of automated SPE with coconut charcoal EPA 521 cartridges. Low method detection limits (MDLs) were achieved, along with a greater facility of the procedure and less dependence on the operator with regard to the methods based on manual SPE. Quality requirements for isotope dilution-based methods were accomplished for most analysed nitrosamines, regarding to trueness (80-120%), method precision (<15%) and MDLs (0.08-1.7 ng/L). Nineteen water samples (16 samples from a drinking water treatment plant {DWTP}, 2 chlorinated samples from a sewage treatment plant {STP} effluent, and 1 chlorinated sample from a reservoir) were analysed. Concentrations of nitrosamines in the STP effluent were 309.4 and 730.2 ng/L, being higher when higher doses of chlorine were applied. N-Nitrosodimethylamine (NDMA) and N-nitrosodiethylamine (NDEA) were the main compounds identified in the STP effluent, and NDEA was detected above 200 ng/L, regulatory level for NDMA in effluents stated in Ontario (Canada). Lower concentrations of nitrosamines were found in the reservoir (20.3 ng/L) and in the DWTP samples (n.d. -28.6 ng/L). NDMA and NDEA were respectively found in the reservoir and in treated and highly chlorinated DWTP samples at concentrations above 10 ng/L (guide value established in different countries). The highest concentrations of nitrosamines were found after chlorination and ozonation processes (ozonated, treated and highly chlorinated water) in DWTP samples.

Colonisation of soilless growing media for tomato by Trichoderma harzianum.

PubMed

De Schutter, B; Aerts, R; Rombouts, L

2001-01-01

An experiment was conducted to evaluate the distribution of T. harzianum in soilless media used in greenhouse growing systems for tomatoes. Growing media based on rockwool and based on coconut fibre were included. The fungus was applied to the roots of the plant by means of a conidial suspension. The upper plant parts were removed from the coconut fibre an the rockwool slab after 10 and 15 weeks respectively. Both the coconut fibre and the rockwool slab were divided into fragments with a width of 10 cm. The coconut fibre medium fragments were again divided in an upper and a lower part. For every fragment the CFU/g was determined by two different methods. In the first method a known volume of growing medium was brought into suspension using a blender and diluted. Appropriate dilutions were plated on a selective medium. In the second method a known volume of growing medium was spread directly upon a selective medium in petri dishes. All dishes were incubated at room temperature and colonies were counted. Results showed that in the case of the coconut fibre growing medium T. harzianum was isolated in every fragment of the substrate. Highest densities were measured at the site of inoculation. In the lower part of the fragments less CFU/g were counted than in the upper part. In the case of the rockwool growing medium, there were several fragments in which no T. harzianum was isolated. The dilution technique demonstrated to be most useful in cases of high density. The direct spreading-method is best applied when low densities are expected. These observations demonstrate that the growing medium based upon coconut fibre is more appropriate for colonisation by T. harzianum. However, densities are higher at the site of inoculation in both tested growing media.

Committee on Diabetes Mellitus Indices of the Japan Society of Clinical Chemistry-recommended reference measurement procedure and reference materials for glycated albumin determination.

PubMed

Takei, Izumi; Hoshino, Tadao; Tominaga, Makoto; Ishibashi, Midori; Kuwa, Katsuhiko; Umemoto, Masao; Tani, Wataru; Okahashi, Mikiko; Yasukawa, Keiko; Kohzuma, Takuji; Sato, Asako

2016-01-01

Glycated albumin is an intermediate glycaemic control marker for which there are several measurement procedures with entirely different reference intervals. We have developed a reference measurement procedure for the purpose of standardizing glycated albumin measurements. The isotope dilution liquid chromatography/tandem mass spectrometry method was developed as a reference measurement procedure for glycated albumin. The stable isotopes of lysine and fructosyl-lysine, which serve as an internal standard, were added to albumin isolated from serum, followed by hydrogenation. After hydrolysis of albumin with hot hydrochloric acid, the liberated lysine and fructosyl-lysine were measured by liquid chromatography/tandem mass spectrometry, and their concentrations were determined from each isotope ratio. The reference materials (JCCRM611) for determining of glycated albumin were prepared from pooled patient blood samples. The isotope dilution-tandem mass spectrometry calibration curve of fructosyl-lysine and lysine showed good linearity (r = 0.999). The inter-assay and intra-assay coefficient of variation values of glycated albumin measurement were 1.2 and 1.4%, respectively. The glycated albumin values of serum in patients with diabetes assessed through the use of this method showed a good relationship with routine measurement procedures (r = 0.997). The relationship of glycated albumin values of the reference material (JCCRM611) between these two methods was the same as the relationship with the patient serum samples. The Committee on Diabetes Mellitus Indices of the Japan Society of Clinical Chemistry recommends the isotope dilution liquid chromatography/tandem mass spectrometry method as a reference measurement procedure, and JCCRM611 as a certified reference material for glycated albumin measurement. In addition, we recommend the traceability system for glycated albumin measurement. © The Author(s) 2015.

In vitro susceptibility of filamentous fungi to copper nanoparticles assessed by rapid XTT colorimetry and agar dilution method.

PubMed

Ghasemian, E; Naghoni, A; Tabaraie, B; Tabaraie, T

2012-12-01

Metal nanoparticles and their uses in various aspects have recently drawn a great deal of attention. One of the major applications is that it can be used as an antimicrobial agent. They can be considered in approaches targeted to decrease the harms caused by microorganisms, specifically fungi, threatening the medical and industrial areas. The aim of this study was to investigate the antifungal activity of synthesized copper nanoparticles (CuNPs) against four filamentous fungi including Alternaria alternata, Aspergillus flavus, Fusarium solani, and Penicillium chrysogenum. Zerovalent copper nanoparticles of mean size 8nm were synthesized by inert gas condensation (IGC) method. The antifungal activity of these synthesized copper nanoparticles was measured against selected fungi by using two different techniques including agar dilution method and XTT reduction assay. The minimal inhibitory concentrations (MICs) for copper nanoparticles by agar dilution method were less or equal to 40mg/L for P. chrysogenum, less or equal to 60mg/L for A. alternata, less or equal to 60mg/L for F. solani, and less or equal to 80mg/L for A. flavus. And also MICs obtained by XTT reduction assay ranged from 40 to 80mg/L. Our data demonstrated that the copper nanoparticles inhibited fungal growth, but the fungal sensitivity to copper nanoparticles varies depending on the fungal species. Therefore, it is advisable that the minimal inhibitory concentrations (MICs) be examined before using these compounds. It is hoped that, in future, copper nanoparticles could replace some antifungal agents, making them applicable to many different medical devices and antimicrobial control system. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Comparative evaluation of the efficacy of a herbal mouthwash and chlorhexidine mouthwash on select periodontal pathogens: An in vitro and ex vivo study

PubMed Central

Pathan, Multazim Muradkhan; Bhat, Kishore Gajanan; Joshi, Vinayak Mahableshwar

2017-01-01

Background: Several herbal mouthwash and herbal extracts have been tested in vitro and in vivo in search of a suitable adjunct to mechanical therapy for long-term use. In this study, we aimed to look at the antimicrobial effect of the herbal mouthwash and chlorhexidine (CHX) mouthwash on select organisms in in vitro test and an ex vivo model. Materials and Methods: The antimicrobial effects were determined against standard strains of bacteria that are involved in different stages of periodontal diseases. The in vitro tests included determination of minimum inhibitory concentration (MIC) using broth dilution and agar diffusion. In the ex vivo part of the study supragingival dental plaque were obtained from 20 periodontally healthy adult volunteers. Descriptive analysis was done for the entire quantitative and qualitative variable recorded. Results: The MIC by broth dilution method found no statistically significant difference between the mouthwashes. The agar dilution method showed CHX was more effective as compared to the herbal mouthwash against standard strains of Streptococcus mutans, Streptococcus sanguinis, and Aggregatibacter actinomycetemcomitans. However, no difference was observed between the mouthwashes for Porphyromonas, Pseudomonas aeruginosa, and Fusobacterium nucleatum. The ex vivo results conclude that none of the selected mouthwashes were statistically significantly different from each other. Conclusion: In the present study, CHX showed higher levels of antimicrobial action than the herbal mouthwash against bacterial species. The results reinforce the earlier findings that the in vitro testing is sensitive to methods and due diligence is needed when extrapolating the data for further use. However, long-term use and in vivo effectiveness against the periopathogens need to be tested in well-planned clinical trials. PMID:29456300

Accurate determination of sulfur in gasoline and related fuel samples using isotope dilution ICP-MS with direct sample injection and microwave-assisted digestion.

PubMed

Heilmann, Jens; Boulyga, Sergei F; Heumann, Klaus G

2004-09-01

Inductively coupled plasma isotope-dilution mass spectrometry (ICP-IDMS) with direct injection of isotope-diluted samples into the plasma, using a direct injection high-efficiency nebulizer (DIHEN), was applied for accurate sulfur determinations in sulfur-free premium gasoline, gas oil, diesel fuel, and heating oil. For direct injection a micro-emulsion consisting of the corresponding organic sample and an aqueous 34S-enriched spike solution with additions of tetrahydronaphthalene and Triton X-100, was prepared. The ICP-MS parameters were optimized with respect to high sulfur ion intensities, low mass-bias values, and high precision of 32S/34S ratio measurements. For validation of the DIHEN-ICP-IDMS method two certified gas oil reference materials (BCR 107 and BCR 672) were analyzed. For comparison a wet-chemical ICP-IDMS method was applied with microwave-assisted digestion using decomposition of samples in a closed quartz vessel inserted into a normal microwave system. The results from both ICP-IDMS methods agree well with the certified values of the reference materials and also with each other for analyses of other samples. However, the standard deviation of DIHEN-ICP-IDMS was about a factor of two higher (5-6% RSD at concentration levels above 100 mircog g(-1)) compared with those of wet-chemical ICP-IDMS, mainly due to inhomogeneities of the micro-emulsion, which causes additional plasma instabilities. Detection limits of 4 and 18 microg g(-1) were obtained for ICP-IDMS in connection with microwave-assisted digestion and DIHEN-ICP-IDMS, respectively, with a sulfur background of the used Milli-Q water as the main limiting factor for both methods.

Diagnostic efficacy of cell block method for vitreoretinal lymphoma.

PubMed

Kase, Satoru; Namba, Kenichi; Iwata, Daiju; Mizuuchi, Kazuomi; Kitaichi, Nobuyoshi; Tagawa, Yoshiaki; Okada-Kanno, Hiromi; Matsuno, Yoshihiro; Ishida, Susumu

2016-03-17

Vitreoretinal lymphoma (VRL) is a life- and sight-threatening disorder. The aim of this study was to analyze the usefulness of the cell block method for diagnosis of VRL. Sixteen eyes in 12 patients with VRL, and 4 eyes in 4 patients with idiopathic uveitis presenting with vitreous opacity were enrolled in this study. Both undiluted vitreous and diluted fluids were isolated during micro-incision vitrectomy. Cell block specimens were prepared in 19 eyes from diluted fluid containing shredding vitreous. These specimens were then submitted for HE staining as well as immunocytological analyses with antibodies against the B-cell marker CD20, the T-cell marker CD3, and cell proliferation marker Ki67. Conventional smear cytology was applied in 14 eyes with VRL using undiluted vitreous samples. The diagnosis of VRL was made based on the results of cytology, concentrations of interleukin (IL)-10 and IL-6 in undiluted vitreous, and immunoglobulin heavy chain gene rearrangement analysis. Atypical lymphoid cells were identified in 14 out of 15 cell block specimens of VRL (positive rate: 93.3 %), but in 5 out of 14 eyes in conventional smear cytology (positive rate: 35.7 %). Atypical lymphoid cells showed immunoreactivity for CD20 and Ki67. Seven cell block specimens were smear cytology-negative and cell block-positive. The cell block method showed no atypical lymphoid cells in any patient with idiopathic uveitis. Cell block specimens using diluted vitreous fluid demonstrated a high diagnostic sensitivity and a low pseudo-positive rate for the cytological diagnosis of VRL. The cell block method contributed to clear differentiation between VRL and idiopathic uveitis with vitreous opacity.

Evaluation of electrical broad bandwidth impedance spectroscopy as a tool for body composition measurement in cows in comparison with body measurements and the deuterium oxide dilution method.

PubMed

Schäff, C T; Pliquett, U; Tuchscherer, A; Pfuhl, R; Görs, S; Metges, C C; Hammon, H M; Kröger-Koch, C

2017-05-01

Body fatness and degree of body fat mobilization in cows vary enormously during their reproduction cycle and influence energy partitioning and metabolic adaptation. The objective of the study was to test bioelectrical impedance spectroscopy (BIS) as a method for predicting fat depot mass (FDM), in living cows. The FDM is defined as the sum of subcutaneous, omental, mesenteric, retroperitoneal, and carcass fat mass. Bioelectrical impedance spectroscopy is compared with the prediction of FDM from the deuterium oxide (DO) dilution method and from body conformation measurements. Charolais × Holstein Friesian (HF; = 18; 30 d in milk) crossbred cows and 2 HF (lactating and nonlactating) cows were assessed by body conformation measurements, BIS, and the DO dilution method. The BCS of cows was a mean of 3.68 (SE 0.64). For the DO dilution method, a bolus of 0.23 g/kg BW DO (60 atom%) was intravenously injected and deuterium (D) enrichment was analyzed in plasma and whey by stabile isotope mass spectrometry, and total body water content was calculated. Impedance measurement was performed using a 4-electrode interface and time domain-based measurement system consisting of a voltage/current converter for applying current stimulus and an amplifier for monitoring voltage across the sensor electrodes. For the BIS, we used complex impedances over three frequency decades that delivers information on intra- and extracellular water and capacity of cell membranes. Impedance data (resistance of extra- and intracellular space, cell membrane capacity, and phase angle) were extracted 1) by simple curve fit to extract the resistance at direct current and high frequency and 2) by using an electrical equivalent circuit. Cows were slaughtered 7 d after BIS and D enrichment measurements and dissected for the measurement of FDM. Multiple linear regression analyses were performed to predict FDM based on data obtained from body conformation measurements, BIS, and D enrichment, and applied methods were evaluated by cross-validation. The FDM varied widely between cows and was correlated to D enrichment in plasma ( = 0.91, < 0.05). Prediction of FDM by body size measurements was less precise ( = 0.84), but FDM prediction was more accurate using D enrichment in plasma ( = 0.90) and BIS ( = 0.99) data. Therefore, both BIS and D enrichment analysis resulted in similarly good predictions of FDM in cows, and we conclude that BIS could have the potential to predict FDM in dairy cows from 40 to 380 kg.

Measuring Dilution of Microbicide Gels with Optical Imaging

PubMed Central

Drake, Tyler K.; Shah, Tejen; Peters, Jennifer J.; Wax, Adam; Katz, David F.

2013-01-01

We present a novel approach for measuring topical microbicide gel dilution using optical imaging. The approach compares gel thickness measurements from fluorimetry and multiplexed low coherence interferometry in order to calculate dilution of a gel. As a microbicide gel becomes diluted at fixed thickness, its mLCI thickness measurement remains constant, while the fluorimetry signal decreases in intensity. The difference between the two measurements is related to the extent of gel dilution. These two optical modalities are implemented in a single endoscopic instrument that enables simultaneous data collection. A preliminary validation study was performed with in vitro placebo gel measurements taken in a controlled test socket. It was found that change in slope of the regression line between fluorimetry and mLCI based measurements indicates dilution. A dilution calibration curve was then generated by repeating the test socket measurements with serial dilutions of placebo gel with vaginal fluid simulant. This methodology can provide valuable dilution information on candidate microbicide products, which could substantially enhance our understanding of their in vivo functioning. PMID:24340006

Structural transformations in diluted micellar and lamellar systems

NASA Astrophysics Data System (ADS)

Zelaya-Rincon, Blanca

The role of dilution by artificial hard water on nanostructures present in body wash samples provided by Procter and Gamble were investigated using time-resolved cryogenic transmission electron microscopy (cryo-TEM). Samples with and without perfume were examined at 10X, 20X, and 50X dilution. Micellar samples transformed to mostly unilamellar vesicles at 50X dilution, in contrast to the micelle to monomer transition seen in typical samples. At lower dilutions, a change in morphology from spherical to wormlike micelles was observed. For lamellar samples, lower dilution ratios show tightly packed multilamellar vesicles, while higher dilution ratios show more dispersed vesicles with less bilayers. Nanostructural transformations upon dilution were attributed to changes in curvature/packing parameters, which occurred due to dilution with hard water and addition of perfume. The systems experience changes in curvature in order to maintain equilibrium. Also, the addition of perfume in the lamellar samples caused an increase in the number of bilayers present in multilamellar vesicles, because of its role in increasing the packing parameter in the system.

Measurement of discharge using tracers

USGS Publications Warehouse

Kilpatrick, Frederick A.; Cobb, Ernest D.

1984-01-01

The development of fluorescent dyes and fluorometers that can measure these dyes at very low concentrations has made dye-dilution methods practical for measuring discharge. These methods are particularly useful for determining discharge under certain flow conditions that are unfavorable for current meter measurements. These include small streams, canals, and pipes where:Turbulence is excessive for current meter measurement but conducive to good mixing.Moving rocks and debris are damaging to any instruments placed in the flow.Cross-sectional areas or velocities are indeterminant or changing.There are some unsteady flows such as exist with storm-runoff events on small streams.The flow is physically inaccessible or unsafe.From a practical standpoint, such measurements are limited primarily to small streams due to excessively long channel mixing lengths required of larger streams. Very good accuracy can be obtained provided:Adequate mixing length and time are allowed.Careful field and laboratory techniques are employed.Dye losses are not significant.This manual describes the slug-injection and constant-rate injection methods of performing tracer-dilution measurements. Emphasis is on the use of fluorescent dyes as tracers and the equipment, field methods, and Laboratory procedures for performing such measurements. The tracer-velocity method is also briefly discussed.

Easy and Fast Method for the Determination of Biogenic Amines in Fish and Fish Products with Liquid Chromatography Coupled to Orbitrap Tandem Mass Spectrometry.

PubMed

Kaufmann, Anton; Maden, Kathryn

2018-03-01

A quantitative method for the determination of biogenic amines was developed. The method is characterized by the virtual absence of sample cleanup and does not require a derivatization reaction. Diluted extracts are centrifuged, filtrated, and directly injected into an ultra-HPLC column, which is coupled to a single-stage high-resolution mass spectrometer (Orbitrap). The chromatography is based on a reversed-phase column and an eluent containing an ion-pairing agent (heptafluorobutyric acid). The high sensitivity of the instrument permits the injection of very diluted extracts, which ensures stable retention times and the virtual absence of signal suppression effects. In addition, the quantification of histamine (a regulated compound) is further aided by the use of an isotopically labeled internal standard. The method was validated for three fish-based matrixes. Both the sample processing and the analytical measurement are very fast; hence, the methodology is ideal for high-throughput work. In addition, the method is significantly more selective than conventional methods (i.e., derivatization followed by LC with UV/fluorescence (FL) detection) for biogenic amines. A comparison showed that LC-UV/FL methods can produce false-positive findings due to coeluting matrix compounds.

An overview of self-consistent methods for fiber-reinforced composites

NASA Technical Reports Server (NTRS)

Gramoll, Kurt C.; Freed, Alan D.; Walker, Kevin P.

1991-01-01

The Walker et al. (1989) self-consistent method to predict both the elastic and the inelastic effective material properties of composites is examined and compared with the results of other self-consistent and elastically based solutions. The elastic part of their method is shown to be identical to other self-consistent methods for non-dilute reinforced composite materials; they are the Hill (1965), Budiansky (1965), and Nemat-Nasser et al. (1982) derivations. A simplified form of the non-dilute self-consistent method is also derived. The predicted, elastic, effective material properties for fiber reinforced material using the Walker method was found to deviate from the elasticity solution for the v sub 31, K sub 12, and mu sub 31 material properties (fiber is in the 3 direction) especially at the larger volume fractions. Also, the prediction for the transverse shear modulus, mu sub 12, exceeds one of the accepted Hashin bounds. Only the longitudinal elastic modulus E sub 33 agrees with the elasticity solution. The differences between the Walker and the elasticity solutions are primarily due to the assumption used in the derivation of the self-consistent method, i.e., the strain fields in the inclusions and the matrix are assumed to remain constant, which is not a correct assumption for a high concentration of inclusions.

Growth on solid media.

PubMed

Elbing, Karen; Brent, Roger

2002-08-01

Detailed protocols are provided for titering and isolating bacterial colonies by serial dilutions, or alternatively by streaking or spreading a plate. Support protocols describe replica plating as well as methods for storing strains as agar stabs or frozen glycerol stocks.

40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

Code of Federal Regulations, 2010 CFR

2010-07-01

... interval. You may use the difference between a diluted exhaust flow meter and a dilution air meter to.... We recommend that you use a diluted exhaust flow meter that meets the specifications in Table 1 of... verification in § 1065.307 and the calibration and verifications in § 1065.340 and § 1065.341. You may use the...

The effect of alkaline pretreatment methods on cellulose structure and accessibility

DOE PAGES

Bali, Garima; Meng, Xianzhi; Deneff, Jacob I.; ...

2014-11-24

The effects of different alkaline pretreatments on cellulose structural features and accessibility are compared and correlated with the enzymatic hydrolysis of Populus. The pretreatments are shown to modify polysaccharides and lignin content to enhance the accessibility for cellulase enzymes. The highest increase in the cellulose accessibility was observed in dilute sodium hydroxide, followed by methods using ammonia soaking and lime (Ca(OH) 2). The biggest increase of cellulose accessibility occurs during the first 10 min of pretreatment, with further increases at a slower rate as severity increases. Low temperature ammonia soaking at longer residence times dissolved a major portion of hemicellulosemore » and exhibited higher cellulose accessibility than high temperature soaking. Moreover, the most significant reduction of degree of polymerization (DP) occurred for dilute sodium hydroxide (NaOH) and ammonia pretreated Populus samples. The study thus identifies important cellulose structural features and relevant parameters related to biomass recalcitrance.« less

Effect of copper and nickel doping on the optical and structural properties of ZnO

NASA Astrophysics Data System (ADS)

Muǧlu, G. Merhan; Sarıtaş, S.; ćakıcı, T.; Şakar, B.; Yıldırım, M.

2017-02-01

The present study is focused on the Cu doped ZnO and Ni doped ZnO dilute magnetic semiconductor thin films. ZnO:Cu and ZnO:Ni thin films were grown by Chemically Spray Pyrolysis (CSP) method on glass substrates. Optical analysis of the films was done spectral absorption and transmittance measurements by UV-Vis double beam spectrophotometer technique. The structure, morphology, topology and elemental analysis of ZnO:Cu and ZnO:Ni dilute magnetic thin films were investigated by X-ray diffraction (XRD), Raman Analysis, field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDX), atomic force microscopy (AFM) techniques, respectively. Also The magnetic properties of the ZnO:Ni thin film was investigated by vibrating sample magnetometer (VSM) method. VSM measurements of ZnO:Ni thin film showed that the ferromagnetic behavior.

Determination of gamma-aminobutyric acid in food matrices by isotope dilution hydrophilic interaction chromatography coupled to mass spectrometry.

PubMed

Zazzeroni, Raniero; Homan, Andrew; Thain, Emma

2009-08-01

The estimation of the dietary intake of gamma-aminobutyric acid (GABA) is dependent upon the knowledge of its concentration values in food matrices. To this end, an isotope dilution liquid chromatography-mass spectrometry method has been developed employing the hydrophilic interaction chromatography technique for analyte separation. This approach enabled accurate quantification of GABA in apple, potato, soybeans, and orange juice without the need of a pre- or post-column derivatization reaction. A selective and precise analytical measurement has been obtained with a triple quadrupole mass spectrometer operating in multiple reaction monitoring using the method of standard additions and GABA-d(6) as an internal standard. The concentrations of GABA found in the matrices tested are 7 microg/g of apple, 342 microg/g of potatoes, 211 microg/g of soybeans, and 344 microg/mL of orange juice.

Ground Motion Studies for Large Future Accelerator

NASA Astrophysics Data System (ADS)

Takeda, Shigeru; Oide, Katsunobu

1997-05-01

The future large accelerator, such as TeV linear collider, should have extremely small emittance to perform the required luminosity. Precise alignment of machine components is essential to prevent emittance dilution. The ground motion spoils alignment of accelerator elements and results in emittance growth. The ground motion in the frequency range of seismic vibration is mostly coherent in the related accelerator. But the incoherent diffusive or Brownian like motion becomes dominant at frequency region less than seismic vibration [1, 2, 3]. Slow ground motion with respect to the machine performance is discussed including the method of tunnel construction. Our experimental results and recent excavated results clarify that application of TBMs is better excavating method than NATM (Drill + Blast) for accelerator tunnel to prevent emittance dilution. ([1] V. Shiltsev, Proc. of IWAA95 Tsukuba, 1995. [2] Shigeru Takeda et al., Proc. of EPAC96, 1996. [3] A. Sery, Proc. of LINAC96, 1996.)

Determination of ruthenium in pharmaceutical compounds by graphite furnace atomic absorption spectroscopy.

PubMed

Jia, Xiujuan; Wang, Tiebang; Bu, Xiaodong; Tu, Qiang; Spencer, Sandra

2006-04-11

A graphite furnace atomic absorption (GFAA) spectrometric method for the determination of ruthenium (Rh) in solid and liquid pharmaceutical compounds has been developed. Samples are dissolved or diluted in dimethyl sulfoxide (DMSO) without any other treatment before they were analyzed by GFAA with a carefully designed heating program to avoid pre-atomization signal loss and to achieve suitable sensitivity. Various inorganic and organic solvents were tested and compared and DMSO was found to be the most suitable. In addition, ruthenium was found to be stable in DMSO for at least 5 days. Spike recoveries ranged from 81 to 100% and the limit of quantitation (LOQ) was determined to be 0.5 microg g(-1) for solid samples or 0.005 microg ml(-1) for liquid samples based a 100-fold dilution. The same set of samples was also analyzed by ICP-MS with a different sample preparation method, and excellent agreement was achieved.

Method of phase space beam dilution utilizing bounded chaos generated by rf phase modulation

DOE PAGES

Pham, Alfonse N.; Lee, S. Y.; Ng, K. Y.

2015-12-10

This paper explores the physics of chaos in a localized phase-space region produced by rf phase modulation applied to a double rf system. The study can be exploited to produce rapid particle bunch broadening exhibiting longitudinal particle distribution uniformity. Hamiltonian models and particle-tracking simulations are introduced to understand the mechanism and applicability of controlled particle diffusion. When phase modulation is applied to the double rf system, regions of localized chaos are produced through the disruption and overlapping of parametric resonant islands and configured to be bounded by well-behaved invariant tori to prevent particle loss. The condition of chaoticity and themore » degree of particle dilution can be controlled by the rf parameters. As a result, the method has applications in alleviating adverse space-charge effects in high-intensity beams, particle bunch distribution uniformization, and industrial radiation-effects experiments.« less

Measurement of lung volumes from supine portable chest radiographs.

PubMed

Ries, A L; Clausen, J L; Friedman, P J

1979-12-01

Lung volumes in supine nonambulatory patients are physiological parameters often difficult to measure with current techniques (plethysmograph, gas dilution). Existing radiographic methods for measuring lung volumes require standard upright chest radiographs. Accordingly, in 31 normal supine adults, we determined helium-dilution functional residual and total lung capacities and measured planimetric lung field areas (LFA) from corresponding portable anteroposterior and lateral radiographs. Low radiation dose methods, which delivered less than 10% of that from standard portable X-ray technique, were utilized. Correlation between lung volume and radiographic LFA was highly significant (r = 0.96, SEE = 10.6%). Multiple-step regressions using height and chest diameter correction factors reduced variance, but weight and radiographic magnification factors did not. In 17 additional subjects studied for validation, the regression equations accurately predicted radiographic lung volume. Thus, this technique can provide accurate and rapid measurement of lung volume in studies involving supine patients.

A novel MLPH variant in dogs with coat colour dilution.

PubMed

Bauer, A; Kehl, A; Jagannathan, V; Leeb, T

2018-02-01

Coat colour dilution may be the result of altered melanosome transport in melanocytes. Loss-of-function variants in the melanophilin gene (MLPH) cause a recessively inherited form of coat colour dilution in many mammalian and avian species including the dog. MLPH corresponds to the D locus in many domestic animals, and recessive alleles at this locus are frequently denoted with d. In this study, we investigated dilute coloured Chow Chows whose coat colour could not be explained by their genotype at the previously known MLPH:c.-22G>A variant. Whole genome sequencing of such a dilute Chow Chow revealed another variant in the MLPH gene: MLPH:c.705G>C. We propose to designate the corresponding mutant alleles at these two variants d 1 and d 2 . We performed an association study in a cohort of 15 dilute and 28 non-dilute Chow Chows. The dilute dogs were all either compound heterozygous d 1 /d 2 or homozygous d 2 /d 2 , whereas the non-dilute dogs carried at least one wildtype allele D. The d 2 allele did not occur in 417 dogs from diverse other breeds. However, when we genotyped a Sloughi family, in which a dilute coloured puppy had been born out of non-dilute parents, we again observed perfect co-segregation of the newly discovered d 2 allele with coat colour dilution. Finally, we identified a blue Thai Ridgeback with the d 1 /d 2 genotype. Thus, our data identify the MLPH:c.705G>C as a variant explaining a second canine dilution allele. Although relatively rare overall, this d 2 allele is segregating in at least three dog breeds, Chow Chows, Sloughis and Thai Ridgebacks. © 2018 Stichting International Foundation for Animal Genetics.

The Effect of Dilution on Microsegregation in AWS ER NiCrMo-14 Alloy Welding Claddings

NASA Astrophysics Data System (ADS)

Miná, Émerson Mendonça; da Silva, Yuri Cruz; Dille, Jean; Silva, Cleiton Carvalho

2016-12-01

Dilution and microsegregation are phenomena inherent to claddings, which, in turn, directly affect their main properties. This study evaluated microsegregation in the fusion zone with different dilution levels. The overlays were welded by the TIG cold wire feed process. Dilution was calculated from the geometric characteristics of the claddings and from the conservation of mass equation using chemical composition measurements. Microsegregation was calculated using energy dispersive X-ray spectroscopy measurements of the dendrites and the chemical composition of the fusion zone. The dilution of the claddings was increased by reducing the wire feed rate. Fe showed potential to be incorporated into the solid phase ( k > 1), and this increased with the increase of dilution. Mo, in turn, was segregated into the liquid phase ( k < 1) and also increased with the increase of dilution. However, Cr and W showed a slight decrease in their partition coefficients ( k) with the increase of dilution.

COMPARISON BETWEEN ELLMAN AND RADIOMETRIC METHODS FOR ASSESSING CHOLINESTERASE (CHE) INHIBITION IN RATS TREATED WITH N-METHYL CARBAMATE INSECTICIDES.

EPA Science Inventory

Carbamylated ChE is unstable and readily reactivates. This reactivation, promoted by increasing temperature and dilution, could have an impact on ex vivo ChE assays by decreasing apparent ChE inhibition. To assess the best method for measuring ChE inhibition in brain and RBCs f...

AN ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) METHOD FOR THE URINARY BIOMONITORING OF 2,4-DICHLOROPHRENOCYACETIC ACID (2,4-D)

EPA Science Inventory

An enzyme-linked immunosorbent assay (ELISA) method was developed to quantitatively measure 2,4-dichlorophenoyacetic acid (2,4-D) in human urine. Samples were diluted (1:5) with phosphate-buffered saline, 0.05% Tween 20, with 0.02% sodium azide, and analyzed by a 96-microwekk pl...

Method development for liquid chromatographic/triple quadrupole mass spectrometric analysis of trace level perfluorocarboxylic acids in articles of commerce

EPA Science Inventory

An analytical method to identify and quantify trace levels of C5 to C12 perfluorocarboxylic acids (PFCAs) in articles of commerce (AOC) is developed and rigorously validated. Solid samples were extracted in methanol, and liquid samples were diluted with a solvent consisting of 60...

Effects of Fuel Composition on EGR Dilution Tolerance in Spark Ignited Engines

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

Szybist, James P

2016-01-01

Fuel-specific differences in exhaust gas recirculation (EGR) dilution tolerance are studied in a modern, direct-injection single-cylinder research engine. A total of 6 model fuel blends are examined at a constant research octane number (RON) of 95 using n-heptane, iso-octane, toluene, and ethanol. Laminar flame speeds for these mixtures, which were calculated two different methods (an energy fraction mixing rule and a detailed kinetic simulation), spanned a range of about 6 cm/s. A constant fueling nominal load of 350 kPa IMEPg at 2000 rpm was operated with varying CA50 from 8-20 CAD aTDCf, and with EGR increasing until a COV ofmore » IMEP of 5% is reached. The results illustrate that flame speed affects EGR dilution tolerance; fuels with increased flame speeds increase EGR tolerance. Specifically, flame speed correlates most closely to the initial flame kernel growth, measured as the time of ignition to 5% mass fraction burned. The effect of the latent heat of vaporization on the flame speed is taken into account for the ethanol-containing fuels. At a 30 vol% blend level, the increased enthalpy of vaporization of ethanol compared to conventional hydrocarbons can decrease the temperature at the time of ignition by a maximum of 15 C, which can account for up to a 3.5 cm/s decrease in flame speed. The ethanol-containing fuels, however, still exhibit a flame speed advantage, and a dilution tolerance advantage over the slower flame-speed fuels. The fuel-specific differences in dilution tolerance are significant at the condition examined, allowing for a 50% relative increase in EGR (4% absolute difference in EGR) at a constant COV of IMEP of 3%.« less

A Computational and Experimental Study of Coflow Laminar Methane/Air Diffusion Flames: Effects of Fuel Dilution, Inlet Velocity, and Gravity

NASA Technical Reports Server (NTRS)

Cao, S.; Ma, B.; Bennett, B. A. V.; Giassi, D.; Stocker, D. P.; Takahashi, F.; Long, M. B.; Smooke, M. D.

2014-01-01

The influences of fuel dilution, inlet velocity, and gravity on the shape and structure of laminar coflow CH4-air diffusion flames were investigated computationally and experimentally. A series of nitrogen-diluted flames measured in the Structure and Liftoff in Combustion Experiment (SLICE) on board the International Space Station was assessed numerically under microgravity (mu g) and normal gravity (1g) conditions with CH4 mole fraction ranging from 0.4 to 1.0 and average inlet velocity ranging from 23 to 90 cm/s. Computationally, the MC-Smooth vorticity-velocity formulation was employed to describe the reactive gaseous mixture, and soot evolution was modeled by sectional aerosol equations. The governing equations and boundary conditions were discretized on a two-dimensional computational domain by finite differences, and the resulting set of fully coupled, strongly nonlinear equations was solved simultaneously at all points using a damped, modified Newton's method. Experimentally, flame shape and soot temperature were determined by flame emission images recorded by a digital color camera. Very good agreement between computation and measurement was obtained, and the conclusions were as follows. (1) Buoyant and nonbuoyant luminous flame lengths are proportional to the mass flow rate of the fuel mixture; computed and measured nonbuoyant flames are noticeably longer than their 1g counterparts; the effect of fuel dilution on flame shape (i.e., flame length and flame radius) is negligible when the flame shape is normalized by the methane flow rate. (2) Buoyancy-induced reduction of the flame radius through radially inward convection near the flame front is demonstrated. (3) Buoyant and nonbuoyant flame structure is mainly controlled by the fuel mass flow rate, and the effects from fuel dilution and inlet velocity are secondary.

Species Associations in a Species-Rich Subtropical Forest Were Not Well-Explained by Stochastic Geometry of Biodiversity

PubMed Central

Wang, Qinggang; Bao, Dachuan; Guo, Yili; Lu, Junmeng; Lu, Zhijun; Xu, Yaozhan; Zhang, Kuihan; Liu, Haibo; Meng, Hongjie; Jiang, Mingxi; Qiao, Xiujuan; Huang, Handong

2014-01-01

The stochastic dilution hypothesis has been proposed to explain species coexistence in species-rich communities. The relative importance of the stochastic dilution effects with respect to other effects such as competition and habitat filtering required to be tested. In this study, using data from a 25-ha species-rich subtropical forest plot with a strong topographic structure at Badagongshan in central China, we analyzed overall species associations and fine-scale species interactions between 2,550 species pairs. The result showed that: (1) the proportion of segregation in overall species association analysis at 2 m neighborhood in this plot followed the prediction of the stochastic dilution hypothesis that segregations should decrease with species richness but that at 10 m neighborhood was higher than the prediction. (2) The proportion of no association type was lower than the expectation of stochastic dilution hypothesis. (3) Fine-scale species interaction analyses using Heterogeneous Poisson processes as null models revealed a high proportion (47%) of significant species effects. However, the assumption of separation of scale of this method was not fully met in this plot with a strong fine-scale topographic structure. We also found that for species within the same families, fine-scale positive species interactions occurred more frequently and negative ones occurred less frequently than expected by chance. These results suggested effects of environmental filtering other than species interaction in this forest. (4) We also found that arbor species showed a much higher proportion of significant fine-scale species interactions (66%) than shrub species (18%). We concluded that the stochastic dilution hypothesis only be partly supported and environmental filtering left discernible spatial signals in the spatial associations between species in this species-rich subtropical forest with a strong topographic structure. PMID:24824996

Species associations in a species-rich subtropical forest were not well-explained by stochastic geometry of biodiversity.

PubMed

Wang, Qinggang; Bao, Dachuan; Guo, Yili; Lu, Junmeng; Lu, Zhijun; Xu, Yaozhan; Zhang, Kuihan; Liu, Haibo; Meng, Hongjie; Jiang, Mingxi; Qiao, Xiujuan; Huang, Handong

2014-01-01

The stochastic dilution hypothesis has been proposed to explain species coexistence in species-rich communities. The relative importance of the stochastic dilution effects with respect to other effects such as competition and habitat filtering required to be tested. In this study, using data from a 25-ha species-rich subtropical forest plot with a strong topographic structure at Badagongshan in central China, we analyzed overall species associations and fine-scale species interactions between 2,550 species pairs. The result showed that: (1) the proportion of segregation in overall species association analysis at 2 m neighborhood in this plot followed the prediction of the stochastic dilution hypothesis that segregations should decrease with species richness but that at 10 m neighborhood was higher than the prediction. (2) The proportion of no association type was lower than the expectation of stochastic dilution hypothesis. (3) Fine-scale species interaction analyses using Heterogeneous Poisson processes as null models revealed a high proportion (47%) of significant species effects. However, the assumption of separation of scale of this method was not fully met in this plot with a strong fine-scale topographic structure. We also found that for species within the same families, fine-scale positive species interactions occurred more frequently and negative ones occurred less frequently than expected by chance. These results suggested effects of environmental filtering other than species interaction in this forest. (4) We also found that arbor species showed a much higher proportion of significant fine-scale species interactions (66%) than shrub species (18%). We concluded that the stochastic dilution hypothesis only be partly supported and environmental filtering left discernible spatial signals in the spatial associations between species in this species-rich subtropical forest with a strong topographic structure.

THE ANTIBACTERIAL PROPERTIES OF SULFUR

PubMed Central

Weld, Julia T.; Gunther, Anne

1947-01-01

1. Saturated solutions of sulfur in alcohol (alcohol-sulfur) when diluted with broth are inhibitory to the growth of various Gram-positive bacteria and to C. hominis. By an arbitrary method of unitage with S. aureus as the test organism, our alcohol-sulfur contains 1,600 to 2,000 units per cc. and one unit contains between 0.24 and 0.34 gamma sulfur. The activity of a preparation is in general directly proportional to its sulfur content. 2. Solutions of sulfur in carbowax (carbowax-sulfur) when diluted with broth are likewise inhibitory to the growth of various Gram-positive bacteria and to C. hominis. When S. aureus is used as test organism, 1 unit contains between 0.1 and 0.2 gamma sulfur. The activity of these preparations is also in general directly proportional to their sulfur content. 3. Carbowax-sulfur when incorporated in agar in 1–500 to 1–2,000 dilution inhibits the growth of various Gram-positive aerobic and anaerobic bacteria, C. hominis, and certain dermatophytes. 4. Our experiments appear to show that both alcohol-sulfur and carbowax-sulfur owe their inhibitory properties to the sulfur particles that are dispersed throughout the medium when these sulfur preparations are diluted with broth. The inhibitory effect of these particles may or may not be due to a combination of the sulfur particles with substances in the medium in which they are suspended. 5. Evidence suggests that the activity of both alcohol-sulfur and carbowax-sulfur is due to sulfur in the same form. The inhibitory effect is characterized by prolonged bacteriostasis with similar activity over a wide range of dilutions. There is no evidence of true bactericidal action even with the highest concentrations used. PMID:19871634

Development of Similar Broth Microdilution Methods to Determine the Antimicrobial Susceptibility of Flavobacterium columnare and F. psychrophilum.

PubMed

Gieseker, Charles M; Crosby, Tina C; Mayer, Tamara D; Bodeis, Sonya M; Stine, Cynthia B

2016-03-01

Flavobacterium columnare and F. psychrophilum are major fish pathogens that cause diseases that may require antimicrobial therapy. Choice of appropriate treatment is dependent upon determining the antimicrobial susceptibility of isolates. Therefore we optimized methods for broth microdilution testing of F. columnare and F. psychrophilum to facilitate standardizing an antimicrobial susceptibility test. We developed adaptations to make reproducible broth inoculums and confirmed the proper incubation time and media composition. We tested the stability of potential quality-control bacteria and compared test results between different operators. Log phase occurred at 48 h for F. columnare and 72-96 h for F. psychrophilum, confirming the test should be incubated at 28°C for approximately 48 h and at 18°C for approximately 96 h, respectively. The most consistent susceptibility results were achieved with plain, 4-g/L, dilute Mueller-Hinton broth supplemented with dilute calcium and magnesium. Supplementing the broth with horse serum did not improve growth. The quality-control strains, Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658, yielded stable minimal inhibitory concentrations (MIC) against all seven antimicrobials tested after 30 passes at 28°C and 15 passes at 18°C. In comparison tests, most MICs of the isolates agreed 100% within one drug dilution for ampicillin, florfenicol, and oxytetracycline. The agreement was lower with the ormetoprim-sulfdimethoxine combination, but there was at least 75% agreement for all but one isolate. These experiments have provided methods to help standardize antimicrobial susceptibility testing of these nutritionally fastidious aquatic bacteria. Received June 24, 2015; accepted October 2, 2015.

A rapid, highly accurate method for quantifying CALR mutant allele burden in persons with myeloproliferative neoplasms.

PubMed

Yao, Qiu-Mei; Zhou, Jiao; Gale, Robert Peter; Li, Jin-Lan; Li, Ling-Di; Li, Ning; Chen, Shan-Shan; Ruan, Guo-Rui

2015-10-01

Calreticulin (CALR) mutations were recently identified in a substantial proportion of persons with essential thrombocythemia (ET) and with primary myelofibrosis (PMF) without JAK2(V617F). Consequently rapid, sensitive, and specific methods to detect and quantify these mutations are needed. We studied samples from 1088 persons with myeloproliferative neoplasms (MPNs) including 421 JAK2(V617F) negative subjects with ET, PMF, polycythemia vera (PV), chronic myeloid leukemia (CML) and hyper-eosinophilic syndrome (HES). Detection of CALR exon 9 mutations was done by PCR amplification followed by fragment length analysis and direct sequencing. Dilution assays were used to determine CALR mutant allele burden. We detected CALR mutations in blood and bone marrow samples from 152 subjects with ET and with PMF but not in samples from normal or persons with PV, CML, or HES. CALR mutant peaks were distinct from wild-type peaks and dilution experiments indicated a sensitivity level of 0.5-5% for a CALR mutant allele in a wild-type background. Diverse types of mutations were detected including deletions, insertions, and complex indels. All mutations were confirmed by direct sequencing. We also used dilution experiments to quantify mutant allele burden. We were able to reproducibly detect mutant allele levels as low 5% (0.5-5%) in a wild-type background. PCR amplification followed by fragment length analysis is a rapid, sensitive, and specific method for screening persons with MPNs for CALR mutations, especially those with ET and PMF and for estimating mutant allele burden.

Effect of quinolones and other antimicrobial agents on cell-associated Legionella pneumophila.

PubMed Central

Havlichek, D; Saravolatz, L; Pohlod, D

1987-01-01

We evaluated the in vitro susceptibility of Legionella pneumophila ATCC 33152 (serogroup I) to 13 antibiotics alone and in combination with rifampin (0.1 mg/liter) by three methods. Extracellular susceptibility was determined by MIC determinations and time kill curves in buffered yeast extract broth, while intracellular susceptibility was determined by peripheral human monocytes in RPMI 1640 culture medium. Antibiotic concentrations equal to or greater than the broth dilution MIC inhibited or killed L. pneumophila by the time kill method, except this was not the case for trimethoprim-sulfamethoxazole. Antibiotic concentrations below the broth dilution MIC did not inhibit Legionella growth. The only antibiotic-rifampin combinations which produced improved killing of L. pneumophila by the time kill method were those in which the logarithmic growth of L. pneumophila occurred during the experiment (rosoxacin, amifloxacin, cinoxacin, trimethoprim-sulfamethoxazole, clindamycin, and doxycycline). Neither direct MICs nor time kill curve assays accurately predicted intracellular L. pneumophila susceptibility. Rifampin, erythromycin, ciprofloxacin, rosoxacin, enoxacin, amifloxacin, gentamicin, clindamycin, and doxycycline all inhibited intracellular L. pneumophila growth at readily achievable concentrations in serum. Cefoxitin and thienamycin showed no inhibition of growth, although they were present extracellularly at concentrations that were 20 to 1,000 times their broth dilution MICs. Clindamycin was the only antibiotic that was able to inhibit intracellular L. pneumophila growth at an extracellular concentration below its MIC. The gentamicin (5 mg/liter)-rifampin combination was the only antibiotic-rifampin combination which demonstrated decreased cell-associated Legionella survival in this model of in vitro susceptibility. PMID:3435101

Determining urea levels in exhaled breath condensate with minimal preparation steps and classic LC-MS.

PubMed

Pitiranggon, Masha; Perzanowski, Matthew S; Kinney, Patrick L; Xu, Dongqun; Chillrud, Steven N; Yan, Beizhan

2014-10-01

Exhaled breath condensate (EBC) provides a relatively easy, non-invasive method for measuring biomarkers of inflammation and oxidative stress in the airways. However, the levels of these biomarkers in EBC are influenced, not only by their levels in lung lining fluid but also by the volume of water vapor that also condenses during EBC collection. For this reason, the use of a biomarker of dilution has been recommended. Urea has been proposed and utilized as a promising dilution biomarker due to its even distribution throughout the body and relatively low volatility. Current EBC urea analytical methods either are not sensitive enough, necessitating large volumes of EBC, or are labor intensive, requiring a derivatization step or other pretreatment. We report here a straightforward and reliable LC-MS approach that we developed that does not require derivatization or large sample volume (∼36 µL). An Acclaim mixed-mode hydrophilic interaction chromatography column was selected because it can produce good peak symmetry and efficiently separate urea from other polar and nonpolar compounds. To achieve a high recovery rate, a slow and incomplete evaporation method was used followed by a solvent-phase exchange. Among EBC samples collected from 28 children, urea levels were found to be highly variable, with a relative standard deviation of 234%, suggesting high variability in dilution of the lung lining fluid component of EBC. The limit of detection was found to be 0.036 µg/mL. Published by Oxford University Press [2013]. This work is written by (a) US Government employee(s) and is in the public domain in the US.

Rapid quantitative analysis of methylphenidate and ritalinic acid in oral fluid by liquid chromatography triple quadrupole mass spectrometry (LC-QqQ-MS).

PubMed

Mulet, Carmen T; Arroyo-Mora, Luis E; Leon, Lorena A; Gnagy, Elizabeth; DeCaprio, Anthony P

2018-06-20

Methylphenidate (MPH), which is metabolized into ritalinic acid (RA), is an amphetamine derivative largely used in the treatment of attention-deficit hyperactivity disorder, a neurological condition commonly diagnosed in early childhood. Ensuring that patients comply with clinical treatment is crucial and compliance is generally monitored in blood or urine specimens which, especially in the case of children, can be challenging to obtain on a repetitive basis. Here we report validation of a specific, non-invasive, and rapid dilute-and-shoot analytical method for the detection and quantitation of MPH and RA in oral fluid (OF). The method is based on liquid chromatography coupled to triple quadrupole MS with electrospray ionization utilizing dynamic MRM mode. Subject OF specimens were collected using a Quantisal™ device, processed, and diluted for analysis with seven-point quadratic calibration curves (weighting of 1/x) using MPH-d 9 and (±)-threo-RA-d 10 as internal standards. QC samples and diluted specimens showed intra- and inter-day bias and imprecision values no greater than ±12%. The LOD and LOQ for MPH were 0.1 and 0.5 ng/mL, respectively, and 0.2 ng/mL and 0.5 ng/mL for RA, respectively, indicating the validity of the method for identification and confirmation at low concentrations. Selectivity was specific for the analytes of interest and matrix effects were minimized through the use of internal standard based quantitation. Copyright © 2018 Elsevier B.V. All rights reserved.

Stable isotope dilution microquantification of creatine metabolites in plasma, whole blood and dried blood spots for pharmacological studies in mouse models of creatine deficiency.

PubMed

Tran, C; Yazdanpanah, M; Kyriakopoulou, L; Levandovskiy, V; Zahid, H; Naufer, A; Isbrandt, D; Schulze, A

2014-09-25

To develop an accurate stable isotope dilution assay for simultaneous quantification of creatine metabolites ornithine, arginine, creatine, creatinine, and guanidinoacetate in very small blood sample volumes to study creatine metabolism in mice. Liquid-chromatography (C18) tandem mass spectrometry with butylation was performed in positive ionization mode. Stable isotope dilution assay with external calibration was applied to three different specimen types, plasma, whole blood and dried blood spot (DBS). Analytical separation, sensitivity, accuracy, and linearity of the assay were adequate. The stable isotope dilution assay in plasma revealed no significant bias to gold standard methods for the respective analytes. Compared to plasma, we observed an overestimate of creatine and creatinine (2- to 5-fold and 1.2- to 2-fold, respectively) in whole-blood and DBS, and an underestimate of arginine (2.5-fold) in DBS. Validation of the assay in mouse models of creatine deficiency revealed plasma creatine metabolite pattern in good accordance with those observed in human GAMT and AGAT deficiency. Single dose intraperitoneal application of ornithine in wild-type mice lead to fast ornithine uptake (Tmax ≤ 10 min) and elimination (T1/2=24 min), and a decline of guanidinoacetate. The assay is fast and reliable to study creatine metabolism and pharmacokinetics in mouse models of creatine deficiency. Copyright © 2014 Elsevier B.V. All rights reserved.

Sensitizing solid state nuclear magnetic resonance of dilute nuclei by spin-diffusion assisted polarization transfers.

PubMed

Lupulescu, Adonis; Frydman, Lucio

2011-10-07

Recent years have witnessed efforts geared at increasing the sensitivity of NMR experiments, by relying on the suitable tailoring and exploitation of relaxation phenomena. These efforts have included the use of paramagnetic agents, enhanced (1)H-(1)H incoherent and coherent transfers processes in 2D liquid state spectroscopy, and homonuclear (13)C-(13)C spin diffusion effects in labeled solids. The present study examines some of the opportunities that could open when exploiting spontaneous (1)H-(1)H spin-diffusion processes, to enhance relaxation and to improve the sensitivity of dilute nuclei in solid state NMR measurements. It is shown that polarization transfer experiments executed under sufficiently fast magic-angle-spinning conditions, enable a selective polarization of the dilute low-γ spins by their immediate neighboring protons. Repolarization of the latter can then occur during the time involved in monitoring the signal emitted by the low-γ nuclei. The basic features involved in the resulting approach, and its potential to improve the effective sensitivity of solid state NMR measurements on dilute nuclei, are analyzed. Experimental tests witness the advantages that could reside from utilizing this kind of approach over conventional cross-polarization processes. These measurements also highlight a number of limitations that will have to be overcome for transforming selective polarization transfers of this kind into analytical methods of choice. © 2011 American Institute of Physics

High throughput workflow for coacervate formation and characterization in shampoo systems.

PubMed

Kalantar, T H; Tucker, C J; Zalusky, A S; Boomgaard, T A; Wilson, B E; Ladika, M; Jordan, S L; Li, W K; Zhang, X; Goh, C G

2007-01-01

Cationic cellulosic polymers find wide utility as benefit agents in shampoo. Deposition of these polymers onto hair has been shown to mend split-ends, improve appearance and wet combing, as well as provide controlled delivery of insoluble actives. The deposition is thought to be enhanced by the formation of a polymer/surfactant complex that phase-separates from the bulk solution upon dilution. A standard characterization method has been developed to characterize the coacervate formation upon dilution, but the test is time and material prohibitive. We have developed a semi-automated high throughput workflow to characterize the coacervate-forming behavior of different shampoo formulations. A procedure that allows testing of real use shampoo dilutions without first formulating a complete shampoo was identified. This procedure was adapted to a Tecan liquid handler by optimizing the parameters for liquid dispensing as well as for mixing. The high throughput workflow enabled preparation and testing of hundreds of formulations with different types and levels of cationic cellulosic polymers and surfactants, and for each formulation a haze diagram was constructed. Optimal formulations and their dilutions that give substantial coacervate formation (determined by haze measurements) were identified. Results from this high throughput workflow were shown to reproduce standard haze and bench-top turbidity measurements, and this workflow has the advantages of using less material and allowing more variables to be tested with significant time savings.

Bioavailability of Cadmium in Inexpensive Jewelry

PubMed Central

Miller, Jennifer; Guinn, Daphne; Pearson, Janna

2011-01-01

Objectives: We evaluated the bioavailability of Cd in 86 components of 57 jewelry items found to contain high levels of Cd (> 10,000 ppm) by X-ray fluorescence (XRF), using extractions that simulate mouthing or swallowing of jewelry items. Methods: We screened jewelry for Cd content by XRF. Bioavailability was measured in two ways. Items were placed in saline solution at 37°C for 6 hr to simulate exposures from mouthing of jewelry items. Items were placed in dilute hydrochloric acid (HCl) at 37°C for 6–96 hr, simulating the worst-case scenario of a child swallowing a jewelry item. Damaged pieces of selected samples were also extracted by both methods to determine the effect of breaching the outer plating on bioavailability. Total Cd content of all items was determined by atomic absorption. Results: The 6-hr saline extraction yielded as much as 2,200 µg Cd, and 24-hr dilute HCl extraction yielded a maximum of > 20,000 µg Cd. Leaching of Cd in dilute HCl increased linearly over 6–96 hr, indicating potential for increasing harm the longer an item remains in the stomach. Damage to jewelry by breaching the outer plating generally, but not always, increased Cd release. Bioavailability did not correlate directly with Cd content. Conclusions: These results indicate the potential for dangerous Cd exposures to children who wear, mouth, or accidentally swallow high-Cd jewelry items. PMID:21377949

Interaction between colloidal particles on an oil-water interface in dilute and dense phases.

PubMed

Parolini, Lucia; Law, Adam D; Maestro, Armando; Buzza, D Martin A; Cicuta, Pietro

2015-05-20

The interaction between micron-sized charged colloidal particles at polar/non-polar liquid interfaces remains surprisingly poorly understood for a relatively simple physical chemistry system. By measuring the pair correlation function g(r) for different densities of polystyrene particles at the decane-water interface, and using a powerful predictor-corrector inversion scheme, effective pair-interaction potentials can be obtained up to fairly high densities, and these reproduce the experimental g(r) in forward simulations, so are self consistent. While at low densities these potentials agree with published dipole-dipole repulsion, measured by various methods, an apparent density dependence and long range attraction are obtained when the density is higher. This condition is thus explored in an alternative fashion, measuring the local mobility of colloids when confined by their neighbors. This method of extracting interaction potentials gives results that are consistent with dipolar repulsion throughout the concentration range, with the same magnitude as in the dilute limit. We are unable to rule out the density dependence based on the experimental accuracy of our data, but we show that incomplete equilibration of the experimental system, which would be possible despite long waiting times due to the very strong repulsions, is a possible cause of artefacts in the inverted potentials. We conclude that to within the precision of these measurements, the dilute pair potential remains valid at high density in this system.

Ariadne's Thread: A Robust Software Solution Leading to Automated Absolute and Relative Quantification of SRM Data.

PubMed

Nasso, Sara; Goetze, Sandra; Martens, Lennart

2015-09-04

Selected reaction monitoring (SRM) MS is a highly selective and sensitive technique to quantify protein abundances in complex biological samples. To enhance the pace of SRM large studies, a validated, robust method to fully automate absolute quantification and to substitute for interactive evaluation would be valuable. To address this demand, we present Ariadne, a Matlab software. To quantify monitored targets, Ariadne exploits metadata imported from the transition lists, and targets can be filtered according to mProphet output. Signal processing and statistical learning approaches are combined to compute peptide quantifications. To robustly estimate absolute abundances, the external calibration curve method is applied, ensuring linearity over the measured dynamic range. Ariadne was benchmarked against mProphet and Skyline by comparing its quantification performance on three different dilution series, featuring either noisy/smooth traces without background or smooth traces with complex background. Results, evaluated as efficiency, linearity, accuracy, and precision of quantification, showed that Ariadne's performance is independent of data smoothness and complex background presence and that Ariadne outperforms mProphet on the noisier data set and improved 2-fold Skyline's accuracy and precision for the lowest abundant dilution with complex background. Remarkably, Ariadne could statistically distinguish from each other all different abundances, discriminating dilutions as low as 0.1 and 0.2 fmol. These results suggest that Ariadne offers reliable and automated analysis of large-scale SRM differential expression studies.

Preparation of fine-particles at cryogenic temperatures

NASA Technical Reports Server (NTRS)

Globus, H.

1970-01-01

Flash freezing process yields gelling agent for use at cryogenic temperatures. Vaporized material, diluted with an inert gas, is injected below the surface of an agitated cryogenic liquid. This method disperses particles of chlorine trifluoride in liquid oxygen difluoride.

REDUCING CHILDREN'S RISK FROM LEAD IN SOIL

EPA Science Inventory

Traditional methods for reducing risk from elevated levels of soil Pb involves removal, covering, or dilution by mixing with uncontaminated soil. Believing that in situ remediation techniques are viable alternatives, the EPA=s National Risk Management Research Laboratory (NRMRL) ...

Real-time cell analysis of the cytotoxicity of the components of orthodontic acrylic materials on gingival fibroblasts.

PubMed

Öztürk, Firat; Malkoc, Siddik; Ersöz, Mustafa; Hakki, Sema S; Bozkurt, Buket S

2011-11-01

The aim of this study was to evaluate the cytotoxicity of 3 orthodontic acrylic materials and 2 manipulation methods. The orthodontic acrylic materials Orthocryl EQ (Dentaurum, Ispringen, Germany), Orthoplast (Vertex Dental, Zeist, The Netherlands), and O-80 (Imicryl, Konya, Turkey) were prepared with 2 polymerization methods (doughing and spray on). Totally, 60 cylinders (5 × 2 mm), fabricated by using a different acrylic and method, were divided into 6 groups. Gingival fibroblasts were isolated from gingival connective tissue of systemically healthy subjects. Materials were incubated in Dulbecco's modified eagle's medium culture medium (Biological Industries, Beit Haemek, Israel) for 72 hours according to ISO 10993-5 standards (surface area to volume ratio of the specimen to cell-culture medium: 3 cm(2)/mL). Gingival fibroblasts were maintained with Dulbecco's modified eagle medium containing 10% fetal bovine serum. A real-time cell analyzer (RT-CES, xCELLigence; Roche Applied Science, Mannheim, Germany, and ACEA Biosciences, San Diego, Calif) was used to evaluate cell survival. After seeding 200 μL of the cell suspensions into the wells (20,000 cells/well) of the E-plate 96, gingival fibroblasts were treated with bioactive components released by the acrylic materials (1/1 and 1/2 dilutions) and monitored every 15 minutes for 121 hours. For the proliferation experiments, the statistical analyses used were 1-way analysis of variance (ANOVA) and Tukey-Kramer multiple comparisons tests. There was no significant difference between the cell indexes of the control and study groups for the 1/1 and 1/2 dilutions at 21 and 32 hours. When evaluated at 68 hours, all 1/2 dilutions of acrylic materials showed statistically insignificant differences (P >0.05) except for Orthoplast (P <0.05). But all acrylic materials were different from the control group in the 1/1 dilutions (P <0.001). At 121 hours, all test groups were significantly different from the untreated control group (P <0.001). The results indicate that the long cycle increased the cytotoxicity of the tested materials, and there was no significant difference between the spray-on and doughing methods on cytotoxicity. Copyright © 2011 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

Determination of low-level (sub-microgram) sulfur concentrations by isotope dilution multi-collector inductively couple plasma mass spectrometry using a 33S spike and internal normalization for mass bias correction.

PubMed

Mann, Jacqueline L; Vocke, Robert D; Kelly, W Robert

2012-05-30

The certification of sulfur (S) in Standard Reference Materials™ by the National Institute of Standards and Technology (NIST) has been exclusively performed using isotope dilution thermal ionization mass spectrometry (ID-TIMS). The ID-TIMS measurement method is limited in its capability for low concentration measurements (<1 µg/g) due to the blank associated with the chemical reduction procedure (≈0.2 S µg). Newly developed materials and applications, such as biofuels made from soy and nanomedicine, pose a challenge to the ID-TIMS technique because of their very low concentrations (<1 µg/g) of S. As described here, a measurement technique with low S blanks is essential for low-level S measurements. An isotope dilution (ID) multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS) method combined with an internal normalization approach for mass bias correction has been used to determine low-level S concentrations in synthetically prepared mixtures using a (33)S-enriched spike material. Dilute sulfur solutions (1 µg S/g) were prepared from NIST SRM 3154 (Sulfate Spectrometric Solution) to test the capability of this technique for measuring very low-level S concentrations. The concentration results for each solution were 0.983 ± 0.004 µg/g (95% CI, n = 2, k = 2), 1.006 ± 0.005 µg/g (95% CI, n = 2, k = 2), and 0.999 ± 0.003 µg/g (95% CI, n = 2, k = 2), in excellent agreement with the gravimetric determination, deviating less than 0.35% and suggesting the technique can yield unbiased and accurate results. The blanks averaged 13 ± 0.0017 ng S (1s). The data results provide a clear indication that the ID-MC-ICP-MS method for the determination of low-level S concentrations is feasible. The more than one order of magnitude reduction of the blanks suggests that it is a better alternative to the ID-TIMS method for very low S materials such as are encountered in biofuels and some biochemical species. Published 2012. This article is a US Government work and is in the public domain in the USA. Published 2012. This article is a US Government work and is in the public domain in the USA.

Antimicrobial susceptibility of porcine Brachyspira hyodysenteriae and Brachyspira pilosicoli isolated in Sweden between 1990 and 2010.

PubMed

Pringle, Märit; Landén, Annica; Unnerstad, Helle Ericsson; Molander, Benedicta; Bengtsson, Björn

2012-09-21

The anaerobic spirochetes Brachyspira hyodysenteriae and Brachyspira pilosicoli cause diarrheal diseases in pigs. Their fastidious nature has hampered standardization of methods for antimicrobial susceptibility testing. For monitoring of antimicrobial susceptibility wild type cutoff values are needed to define where the wild type distribution of MICs ends and no approved cutoffs are available for Brachyspira spp. In this study antimicrobial susceptibility data for both species (in total 906 isolates) were compiled and analyzed and wild type cut off values for B. hyodysenteriae proposed. The MICs of tiamulin, valnemulin, tylosin, tylvalosin, doxycycline and lincomycin were determined by broth dilution in brain heart infusion broth supplemented with 10% fetal calf serum. The compiled MICs from the broth dilution tests of the B. hyodysenteriae type strain, B78T (ATCC® 27164T), showed that the method yields reproducible results. In an international perspective the frequencies of isolates with decreased antimicrobial susceptibility were low among both B. hyodysenteriae and B. pilosicoli. However, in B. pilosicoli a constant level of 10-15% isolates with tiamulin MICs >4 μg/ml was detected between 2002 and 2010 and in B. hyodysenteriae a gradual increase in tiamulin MICs was seen between 1990 and 2003 although this increase has ceased during the last years. The wild type cutoff values proposed for B. hyodysenteriae are: tiamulin >0.25 μg/ml, valnemulin >0.125 μg/ml, tylosin >16 μg/ml, tylvalosin >1 μg/ml, lincomycin >1 μg/ml and doxycycline >0.5 μg/ml. The broth dilution method used in this study has over the years generated tightly grouped MIC populations for the field isolates and reproducible results for the control strain B78T and is therefore a suitable antimicrobial susceptibility test method for monitoring of Brachyspira spp. Here we propose wild type cutoff values for six antimicrobial agents for B. hyodysenteriae tested by broth dilution based on MIC distributions and the current knowledge on mechanisms of resistance in this species. There are few studies on antimicrobial resistance mechanisms and MIC distributions in B. pilosicoli but to some extent the cutoff values proposed for B. hyodysenteriae may be applicable also for monitoring of antimicrobial susceptibility in B. pilosicoli.

Nanoparticles and Ocean Optics

DTIC Science & Technology

2005-09-30

ANSI Std Z39-18 control infection by the dilution process. Subsequent dilution experiments used eukaryotic hosts ( Emiliania huxleyi) and its...Results of dilution experiment with Emiliania huxleyi (strain 88E) and its virus. In this experiment, infection rate is controlled by the dilution

Determination of refractive index, size, and concentration of nonabsorbing colloidal nanoparticles from measurements of the complex effective refractive index.

PubMed

Márquez-Islas, Roberto; Sánchez-Pérez, Celia; García-Valenzuela, Augusto

2014-02-01

We describe a method for obtaining the refractive index (RI), size, and concentration of nonabsorbing nanoparticles in suspension from relatively simple optical measurements. The method requires measuring the complex effective RI of two dilute suspensions of the particles in liquids of different refractive indices. We describe the theoretical basis of the proposed method and provide experimental results validating the procedure.

Technical note: Evaluating nuclear magnetic resonance spectroscopy for determining body composition in Holstein dairy calves using deuterium oxide dilution methods.

PubMed

Chapman, C E; Wilkinson, P Stone; Murphy, M R; Erickson, P S

2017-04-01

Deuterium oxide (D 2 O) dilution methods have been used to assess body composition in live animals. Estimated body water content can be used to predict body fat and protein, and thus, the amount of energy reserves. It is an alternative method to direct chemical analysis and considered a noninvasive technique that is economical and repeatable. Deuterium oxide use is considered easy, safe, and accurate; however, the traditional methods of analyzing D 2 O are expensive, tedious, and time consuming. The objective of this study was to evaluate the potential for using nuclear magnetic resonance spectroscopy (NMR) to determine body composition in Holstein dairy heifers. Nuclear magnetic resonance is less expensive and requires minutes to calculate the percentage of D 2 O in the blood. This study used 24 newborn dairy heifer calves blocked by birth and randomly assigned to 1 of 3 treatments: (1) 446 g dry matter (DM) of a conventional milk replacer (MR) [CON; 20% crude protein (CP), 20% fat], (2) 669 g DM of a moderately high protein MR (MOD; 26% CP, 18% fat), or (3) 892 g DM of a moderately high protein MR (aggressive, AGG; 26% CP, 18% fat). All calves had free-choice access to starter and water. Both MR and starter were medicated with decoquinate. During weaning (d 43 to 49), the morning MR feeding ceased. On d 50, all MR feedings ended but starter and water intakes were continuously recorded until d 56. When calves were 50 d of age, a baseline blood sample was taken followed by injection of 300 mg of D 2 O/kg of body weight in sterile physiological saline (0.9%). The syringes containing the D 2 O in physiological saline were weighed before and after administration to record the actual dose of D 2 O injected gravimetrically. After injection, the D 2 O was allowed to equilibrate with body water for 1 h. Six blood samples were taken over 6 d (1/d) at 1630 h to estimate the dilution of the tracer. The plasma was aspirated and stored at -20°C until further D 2 O analysis. This new method was validated using 4 calf plasma samples that were sent to an outside laboratory for measurement using an independent validation method. We detected no differences in total body water, protein, fat, or mineral content in calves fed CON, MOD, or AGG; however, results demonstrated that the D 2 O dilution technique and analysis by NMR is an appropriate and easy method to estimate water, protein, ash, and fat in young heifers. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Comparison of a gel microcolumn assay with the conventional tube test for red blood cell alloantibody titration.

PubMed

Finck, Rachel; Lui-Deguzman, Carrie; Teng, Shih-Mao; Davis, Rebecca; Yuan, Shan

2013-04-01

Titration is a semiquantitative method used to estimate red blood cell (RBC) alloantibody reactivity. The conventional tube test (CTT) technique is the traditional method for performing titration studies. The gel microcolumn assay (GMA) is also a sensitive method to detect RBC alloantibodies. The aim of this study was to compare a GMA with the CTT technique in the performance of Rh and K alloantibody titration. Patient serum samples that contained an RBC alloantibody with a singular specificity were identified by routine blood bank workflow. Parallel titration studies were performed on these samples by both the CTT method and a GMA (ID-Micro Typing System anti-IgG gel card, Micro Typing Systems, Inc., an Ortho-Clinical Diagnostics Company). Forty-eight samples were included, including 11 anti-D, five anti-c, 13 anti-E, one anti-C, three anti-e, and 15 anti-K. Overall, the two methods generated identical results in 21 of 48 samples. For 42 samples (87.5%) the two methods generated results that were within one serial dilution, and for the remaining six samples, results were within two dilutions. GMA systems may perform comparably to the CTT in titrating alloantibodies to Rh and Kell antigens. © 2012 American Association of Blood Banks.