Dilution-to-extinction cultivation of leaf-inhabiting endophytic fungi in beech (Fagus sylvatica L.)--different cultivation techniques influence fungal biodiversity assessment.

PubMed

Unterseher, Martin; Schnittler, Martin

2009-05-01

Two cultivation-based isolation techniques - the incubation of leaf fragments (fragment plating) and dilution-to-extinction culturing on malt extract agar - were compared for recovery of foliar endophytic fungi from Fagus sylvatica near Greifswald, north-east Germany. Morphological-anatomical characters of vegetative and sporulating cultures and ITS sequences were used to assign morphotypes and taxonomic information to the isolates. Data analysis included species-accumulation curves, richness estimators, multivariate statistics and null model testing. Fragment plating and extinction culturing were significantly complementary with regard to species composition, because around two-thirds of the 35 fungal taxa were isolated with only one of the two cultivation techniques. The difference in outcomes highlights the need for caution in assessing fungal biodiversity based upon single isolation techniques. The efficiency of cultivation-based studies of fungal endophytes was significantly increased with the combination of the two isolation methods and estimations of species richness, when compared with a 20-years old reference study, which needed three times more isolates with fragment plating to attain the same species richness. Intensified testing and optimisation of extinction culturing in endophyte research is advocated.

Vibration Mitigation for a Cryogen-Free Dilution Refrigerator for the AMoRE-Pilot Experiment

NASA Astrophysics Data System (ADS)

Lee, C.; Jo, H. S.; Kang, C. S.; Kim, G. B.; Kim, I.; Kim, Y. H.; Lee, H. J.; So, J. H.

2018-06-01

The Advanced Mo-based Rare process Experiment utilizes a cryogen-free dilution refrigerator to operate its low-temperature detectors. Mechanical vibration originating from its pulse tube refrigerator can affect the detector performance. A mechanical filter system has been installed between the 4K and still plates with eddy current dampers in addition to a spring-loaded damping system previously installed below the mixing chamber plate of the cryostat. The filters significantly mitigated vibrations and improved the detector signals.

Probe colorimeter for quantitating enzyme-linked immunosorbent assays and other colorimetric assays performed with microplates.

PubMed Central

Ackerman, S B; Kelley, E A

1983-01-01

The performance of a fiberoptic probe colorimeter (model PC800; Brinkmann Instruments, Inc., Westbury, N.Y.) for quantitating enzymatic or colorimetric assays in 96-well microtiter plates was compared with the performances of a spectrophotometer (model 240; Gilford Instrument Laboratories, Inc., Oberlin, Ohio) and a commercially available enzyme immunoassay reader (model MR590; Dynatech Laboratories, Inc., Alexandria, Va.). Alkaline phosphatase-p-nitrophenyl phosphate in 3 M NaOH was used as the chromophore source. Six types of plates were evaluated for use with the probe colorimeter; they generated reproducibility values (100% coefficient of variation) ranging from 91 to 98% when one individual made 24 independent measurements on the same dilution of chromophore on each plate. Eleven individuals each performed 24 measurements with the colorimeter on either a visually light (absorbance of 0.10 at 420 nm) or a dark (absorbance of 0.80 at 420 nm) dilution of chromophore; reproducibilities averaged 87% for the light dilution and 97% for the dark dilution. When one individual measured the same chromophore sample at least 20 times in the colorimeter, in the spectrophotometer or in the enzyme immunoassay reader, reproducibility for each instrument was greater than 99%. Measurements of a dilution series of chromophore in a fixed volume indicated that the optical responses of each instrument were linear in a range of 0.05 to 1.10 absorbance units. Images PMID:6341399

Probe colorimeter for quantitating enzyme-linked immunosorbent assays and other colorimetric assays performed with microplates.

PubMed

Ackerman, S B; Kelley, E A

1983-03-01

The performance of a fiberoptic probe colorimeter (model PC800; Brinkmann Instruments, Inc., Westbury, N.Y.) for quantitating enzymatic or colorimetric assays in 96-well microtiter plates was compared with the performances of a spectrophotometer (model 240; Gilford Instrument Laboratories, Inc., Oberlin, Ohio) and a commercially available enzyme immunoassay reader (model MR590; Dynatech Laboratories, Inc., Alexandria, Va.). Alkaline phosphatase-p-nitrophenyl phosphate in 3 M NaOH was used as the chromophore source. Six types of plates were evaluated for use with the probe colorimeter; they generated reproducibility values (100% coefficient of variation) ranging from 91 to 98% when one individual made 24 independent measurements on the same dilution of chromophore on each plate. Eleven individuals each performed 24 measurements with the colorimeter on either a visually light (absorbance of 0.10 at 420 nm) or a dark (absorbance of 0.80 at 420 nm) dilution of chromophore; reproducibilities averaged 87% for the light dilution and 97% for the dark dilution. When one individual measured the same chromophore sample at least 20 times in the colorimeter, in the spectrophotometer or in the enzyme immunoassay reader, reproducibility for each instrument was greater than 99%. Measurements of a dilution series of chromophore in a fixed volume indicated that the optical responses of each instrument were linear in a range of 0.05 to 1.10 absorbance units.

Evaluation of different selective media and culturing techniques for the quantification of Campylobacter ssp. from broiler litter.

PubMed

Kiess, A S; Parker, H M; McDaniel, C D

2010-08-01

Poultry is a major reservoir for Campylobacter, the leading cause of foodborne illness in the United States, but how broilers become initially colonized is still under debate. Broiler litter is a potential source, but the best technique for quantifying Campylobacter from litter is still unknown. Therefore, our objectives were to determine if certain media are more selective for quantifying Campylobacter and if enrichment allows for the detection of stressed or viable but nonculturable cells from broiler litter samples. In this trial, 5 media and 2 culturing techniques were used to enumerate Campylobacter from broiler litter. The media used were campy-Line agar (CLA), campy-cefex agar (CCA), modified CCA, Campylobacter agar plates (CAP), and modified charcoal cefoperazone deoxycholate agar. Litter samples were obtained from a commercial broiler house. Each sample was equally divided and diluted 10-fold into peptone, for direct plating, or 4-fold into Campylobacter enrichment broth. Samples diluted in peptone were direct-plated onto each media and incubated under microaerophilic conditions for 48 h at 42 degrees C. Samples diluted in enrichment broth were incubated under the same conditions for 24 h, then further diluted to 10-fold before plating. Plates from enriched samples were incubated for an additional 24 h after plating. After incubation, all plates (direct and enriched) were counted and presumptive positive colonies were confirmed using a Campylobacter latex agglutination kit. Results indicated that there was no difference in the ability of any of the selective media tested to grow Campylobacter. Direct-plated samples had a higher Campylobacter isolation rate compared with enriched samples. The CLA and CAP were able to suppress total bacterial growth better than modified charcoal cefoperazone deoxycholate, modified CCA, and CCA. The CLA and CAP were the only media able to detect total bacterial population shifts over time. In conclusion, it is important before making a final decision on a selective medium to consider the medium's ability to suppress total bacterial growth as well as isolate Campylobacter.

Comparison of an automated Most Probable Number (MPN) technique to traditional plating methods for estimating populations of total aerobes, coliforms and E. coli associated with freshly processed broiler chickens

USDA-ARS?s Scientific Manuscript database

Traditional microbiological techniques for estimating populations of viable bacteria can be laborious and time consuming. The Most Probable Number (MPN) technique is especially tedious as multiple series of tubes must be inoculated at several different dilutions. Recently, an instrument (TEMPOTM) ...

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.

Monitoring Microbial Numbers in Food by Density Centrifugation

PubMed Central

Basel, Richard M.; Richter, Edward R.; Banwart, George J.

1983-01-01

Some foods contain low numbers of microbes that may be difficult to enumerate by the plate count method due to small food particles that interfere with the counting of colonies. Ludox colloidal silicon was coated with reducing agents to produce a nontoxic density material. Food homogenates were applied to a layered 10 and 80% mixture of modified Ludox and centrifuged at low speed. The top and bottom of the tube contained the food material, and the Ludox-containing portion was evaluated by conventional pour plate techniques. Plate counts of the Ludox mixture agreed with plate counts of the food homogenate alone. The absence of small food particles from pour plates resulted in a plate that was more easily read than pour plates of the homogenate alone. Modified Ludox was evaluated for its effect on bacteria at 4°C during a 24-h incubation period. No inhibition was observed. This method is applicable to food products, such as doughnuts, spices, tomato products, and meat, in which small food particles often interfere with routine plate counts or low dilution may inhibit colony formation. Inhibitory substances can be removed from spices, resulting in higher counts. Ludox is more economical than similar products, such as Percoll. Modified Ludox is easily rendered nontoxic by the addition of common laboratory reagents. In addition, the mixture is compatible with microbiological media. PMID:6303217

Differential bacteriophage mortality on exposure to copper.

PubMed

Li, Jinyu; Dennehy, John J

2011-10-01

Many studies report that copper can be used to control microbial growth, including that of viruses. We determined the rates of copper-mediated inactivation for a wide range of bacteriophages. We used two methods to test the effect of copper on bacteriophage survival. One method involved placing small volumes of bacteriophage lysate on copper and stainless steel coupons. Following exposure, metal coupons were rinsed with lysogeny broth, and the resulting fluid was serially diluted and plated on agar with the corresponding bacterial host. The second method involved adding copper sulfate (CuSO(4)) to bacteriophage lysates to a final concentration of 5 mM. Aliquots were removed from the mixture, serially diluted, and plated with the appropriate bacterial host. Significant mortality was observed among the double-stranded RNA (dsRNA) bacteriophages Φ6 and Φ8, the single-stranded RNA (ssRNA) bacteriophage PP7, the ssDNA bacteriophage ΦX174, and the dsDNA bacteriophage PM2. However, the dsDNA bacteriophages PRD1, T4, and λ were relatively unaffected by copper. Interestingly, lipid-containing bacteriophages were most susceptible to copper toxicity. In addition, in the first experimental method, the pattern of bacteriophage Φ6 survival over time showed a plateau in mortality after lysates dried out. This finding suggests that copper's effect on bacteriophage is mediated by the presence of water.

Machine for Automatic Bacteriological Pour Plate Preparation

PubMed Central

Sharpe, A. N.; Biggs, D. R.; Oliver, R. J.

1972-01-01

A fully automatic system for preparing poured plates for bacteriological analyses has been constructed and tested. The machine can make decimal dilutions of bacterial suspensions, dispense measured amounts into petri dishes, add molten agar, mix the dish contents, and label the dishes with sample and dilution numbers at the rate of 2,000 dishes per 8-hr day. In addition, the machine can be programmed to select different media so that plates for different types of bacteriological analysis may be made automatically from the same sample. The machine uses only the components of the media and sterile polystyrene petri dishes; requirements for all other materials, such as sterile pipettes and capped bottles of diluents and agar, are eliminated. Images PMID:4560475

Enumeration of the contaminating bacterial microbiota in unfermented pasteurized milks enriched with probiotic bacteria.

PubMed

Champagne, C P; Raymond, Y; Gonthier, J; Audet, P

2009-04-01

Pasteurized and unfermented milks supplemented with probiotic bacteria are appearing on the market. It then becomes a challenge to ascertain the undesirable contamination microbiota in the presence of a largely superior population of probiotic bacteria. A method to enumerate the contaminating microbial microbiota in such probiotic-enriched milks was developed. The probiotic cultures, Lactobacillus rhamnosus Lb-Immuni-T and Bifidobacterium animalis subsp. lactis BB-12(R), were added to a pasteurized unfermented milk to reach a minimum of 1 billion CFU per 250 mL portion, as ascertained by plating on de Man - Rogosa - Sharpe (MRS) agar in anaerobic conditions. No growth of B. animalis subsp. lactis BB-12 was noted on plate count agar (PCA) or Petrifilm plates, and the presence of this culture did not affect standard plate counts (SPC) of contaminating bacteria. However, L. rhamnosus formed colonies on PCA and Petrifilm plates. Attempts were thus made to inhibit the growth of the probiotic lactobacilli in PCA. The addition of 2% sodium phosphate (SP) or 5% glycerophosphate (GP) inhibited the growth of the lactobacilli in broths, but pin-point colonies of L. rhamnosus Lb-Immuni-T nevertheless appeared on PCA supplemented with phosphates. SPC could be obtained on PCA + 2% SP by only counting the large colonies, but this resulted in a significant (4.4 fold) underestimation of SPC values. On Petrifilm AC, at dilutions 0 to 2, all colonies were considered as being contaminants, while at dilutions 3 and 4, only large colonies were counted for SPC determinations. There was a direct correlation (R2 = 0.99) between SPC values with Petrifilm in uninoculated milks and those obtained on probiotic-enriched milks. The high correlation obtained over the 102 to 106 CFU/mL range of SPC values show that this Petrifilm method is appropriate to evaluate the microbiological quality of pasteurized milks enriched with L. rhamnosus Lb-Immuni-T and B. animalis subsp. lactis BB-12.

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.

Biocompatibility of hydroxyapatite coatings deposited by pulse electrodeposition technique on the Nitinol superelastic alloy.

PubMed

Marashi-Najafi, F; Khalil-Allafi, J; Etminanfar, M R

2017-07-01

The present study deals with pulse electrochemical deposition of HA on NiTi alloy and in vitro evaluation of coatings. At first step, a thermo-chemical surface modification process was applied to control the Ni release of the alloy. The electrochemical deposition of CaP coatings was examined at both dilute and concentrated solutions. The morphology and the composition of coatings were studied using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Plate like and needle like morphologies were formed for dilute and concentrated solution respectively and HA phase was formed by increasing the pulse current density for both electrolyte. The thickness of the samples was measured using cross sectioning technique. Fibroblast cell culture test on the coated samples revealed that the HA coating obtained by dilute solution shows the best biocompatibility. Also, MTT assay showed the highest cell density and cell proliferation after 5days for the HA coating of dilute solution. The contact angle of samples was measured and the coated samples showed a hydrophilic surface. Soaking the sample in SBF revealed that the crystallization rate of calcium-phosphate compounds is higher on the plate like HA coating as compared to the needle like morphology. The P release of the HA coated samples was measured in a physiological saline solution and the results show that the ions releasing in the plate like coating are less than the needle like coating. It seems that the stability of the plate like coating in biological environments is responsible for the better biocompatibility of the coating. Copyright © 2017 Elsevier B.V. All rights reserved.

Analysis of polonium-210 in food products and bioassay samples by isotope-dilution alpha spectrometry.

PubMed

Lin, Zhichao; Wu, Zhongyu

2009-05-01

A rapid and reliable radiochemical method coupled with a simple and compact plating apparatus was developed, validated, and applied for the analysis of (210)Po in variety of food products and bioassay samples. The method performance characteristics, including accuracy, precision, robustness, and specificity, were evaluated along with a detailed measurement uncertainty analysis. With high Po recovery, improved energy resolution, and effective removal of interfering elements by chromatographic extraction, the overall method accuracy was determined to be better than 5% with measurement precision of 10%, at 95% confidence level.

Growth in liquid or solid media.

PubMed

Elbing, K L; Brent, R

2001-05-01

This appendix presents basic procedures for growing overnight (and larger) cultures, monitoring growth, and titering and isolating bacterial cultures by serial dilution. In addition, protocols are provided for isolating single colonies by streaking and spreading a plate. Replica plating and strain storage/retrieval is also detailed.

Heat transfer optimization for air-mist cooling between a stack of parallel plates

NASA Astrophysics Data System (ADS)

Issa, Roy J.

2010-06-01

A theoretical model is developed to predict the upper limit heat transfer between a stack of parallel plates subject to multiphase cooling by air-mist flow. The model predicts the optimal separation distance between the plates based on the development of the boundary layers for small and large separation distances, and for dilute mist conditions. Simulation results show the optimal separation distance to be strongly dependent on the liquid-to-air mass flow rate loading ratio, and reach a limit for a critical loading. For these dilute spray conditions, complete evaporation of the droplets takes place. Simulation results also show the optimal separation distance decreases with the increase in the mist flow rate. The proposed theoretical model shall lead to a better understanding of the design of fins spacing in heat exchangers where multiphase spray cooling is used.

In Vitro Effect of Zingiber officinale Extract on Growth of Streptococcus mutans and Streptococcus sanguinis.

PubMed

Azizi, Arash; Aghayan, Shabnam; Zaker, Saeed; Shakeri, Mahdieh; Entezari, Navid; Lawaf, Shirin

2015-01-01

Background and Objectives. Tooth decay is an infectious disease of microbial origin. Considering the increasing prevalence of antibiotic resistance due to their overuse and also their side effects, medicinal plants are now considered for use against bacterial infections. This study aimed to assess the effects of different concentrations of Zingiber officinale extract on proliferation of Streptococcus mutans and Streptococcus sanguinis in vitro. Materials and Methods. In this experimental study, serial dilutions of the extract were prepared in two sets of 10 test tubes for each bacterium (total of 20). Standard amounts of bacterial suspension were added; 100ƛ of each tube was cultured on prepared solid agar plates and incubated at 37°C for 24 hours. Serial dilutions of the extract were prepared in another 20 tubes and 100ƛ of each tube was added to blood agar culture medium while being prepared. The mixture was transferred to the plates. The bacteria were inoculated on plates and incubated as described. Results. The minimum inhibitory concentration (MIC) was 0.02 mg/mL for S. mutans and 0.3 mg/mL for S. sanguinis. The minimum bactericidal concentration (MBC) was 0.04 mg for S. mutans and 0.6 mg for S. sanguinis. Conclusion. Zingiber officinale extract has significant antibacterial activity against S. mutans and S. sanguinis cariogenic microorganisms.

In Vitro Effect of Zingiber officinale Extract on Growth of Streptococcus mutans and Streptococcus sanguinis

PubMed Central

Azizi, Arash; Aghayan, Shabnam; Zaker, Saeed; Shakeri, Mahdieh; Entezari, Navid; Lawaf, Shirin

2015-01-01

Background and Objectives. Tooth decay is an infectious disease of microbial origin. Considering the increasing prevalence of antibiotic resistance due to their overuse and also their side effects, medicinal plants are now considered for use against bacterial infections. This study aimed to assess the effects of different concentrations of Zingiber officinale extract on proliferation of Streptococcus mutans and Streptococcus sanguinis in vitro. Materials and Methods. In this experimental study, serial dilutions of the extract were prepared in two sets of 10 test tubes for each bacterium (total of 20). Standard amounts of bacterial suspension were added; 100ƛ of each tube was cultured on prepared solid agar plates and incubated at 37°C for 24 hours. Serial dilutions of the extract were prepared in another 20 tubes and 100ƛ of each tube was added to blood agar culture medium while being prepared. The mixture was transferred to the plates. The bacteria were inoculated on plates and incubated as described. Results. The minimum inhibitory concentration (MIC) was 0.02 mg/mL for S. mutans and 0.3 mg/mL for S. sanguinis. The minimum bactericidal concentration (MBC) was 0.04 mg for S. mutans and 0.6 mg for S. sanguinis. Conclusion. Zingiber officinale extract has significant antibacterial activity against S. mutans and S. sanguinis cariogenic microorganisms. PMID:26347778

[A comparison between 2 different methods for calculating the percentage of anaerobic microorganisms in the subgingival microbial flora].

PubMed

Petti, S; Renzini, G

1994-03-01

The percentage of anaerobic micro-organisms in the subgingival microflora represents a simple microbiological index which not only refers to the state but also the risks of periodontal health. The present study aimed to compare two different methods of calculating this index. The study was performed in 45 subjects with moderate gingivitis provoked by the previous application of dental fixtures anchored to both arches. A sample of subgingival microflora was collected from each patient at the level of the vestibular gingival sulcus of the first upper right molar. This was then vortexed, diluted and inoculated in three series of plates. It was chosen to use Walker's culture medium. The total bacterial count was evaluated by incubating the first series of plates in anaerobiosis; the anaerobic bacterial was calculated by subtracting from the total the of facultative aerobic-anaerobic micro-organisms, which in turn was obtained using two methods: the first (method AE) consisted of incubating another series of plates in aerobiosis; the second (method M) involved incubating the last series of plates in anaerobiosis, and adding metronidazole to the culture medium in a solution of 2.5 mg/l. The plates were then kept at 37 degrees C for seven days. The mean percentage of anaerobic microorganisms, given by the percentage ratio between anaerobic and total, relating to the 45 cases studied, was as follows: using method AE: 57.8 +/- 26.3%, and using method M: 40.2 +/- 27.2%. Both figures come close to that proposed and calculated using a much more sophisticated method by Slots, namely 41.5 +/- 19.2% in the event of gingivitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Plating isolation of various catalase-negative microorganisms from soil

NASA Technical Reports Server (NTRS)

Labeda, D. P.; Hunt, C. M.; Casida, L. E., Jr.

1974-01-01

A unique plating procedure was developed that allows isolation, but not enumeration, of representatives of the catalase-negative soil microflora. The numbers recovered, however, are low as compared to the numbers recovered when the modified dilution-to-extinction isolation procedure is used. The latter procedure provides prolonged inoculation in sealed tubes containing a nutritionally rich broth medium over small submerged agar slants. In contrast, the plating procedure utilizes nutritionally minimal media and the shorter incubations mandated by the inherent problems associated with plating.

40 CFR 1066.110 - Equipment specifications for emission sampling systems.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., such as orifice plates or fins, to achieve good mixing; this may be necessary for good mixing if Re# is... condensation. (v) If you choose to dilute the exhaust by using a remote mix tee, which dilutes the exhaust at... polymer-based materials. (D) Use good engineering judgment to ensure that the materials you choose do not...

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.

Antimicrobial susceptibility testing of Mycobacterium tuberculosis complex for first and second line drugs by broth dilution in a microtiter plate format.

PubMed

Hall, Leslie; Jude, Kurt P; Clark, Shirley L; Wengenack, Nancy L

2011-06-24

The rapid detection of antimicrobial resistance is important in the effort to control the increase in resistant Mycobacterium tuberculosis (Mtb). Antimicrobial susceptibility testing (AST) of Mtb has traditionally been performed by the agar method of proportion or by macrobroth testing on an instrument such as the BACTEC (Becton Dickinson, Sparks, MD), VersaTREK (TREK Diagnostics, Cleveland, OH) or BacT/ALERT (bioMérieux, Hazelwood, MO). The agar proportion method, while considered the "gold" standard of AST, is labor intensive and requires calculation of resistance by performing colony counts on drug-containing agar as compared to drug-free agar. If there is ≥1% growth on the drug-containing medium as compared to drug-free medium, the organism is considered resistant to that drug. The macrobroth methods require instrumentation and test break point ("critical") drug concentrations for the first line drugs (isoniazid, ethambutol, rifampin, and pyrazinamide). The method described here is commercially available in a 96 well microtiter plate format [MYCOTB (TREK Diagnostics)] and contains increasing concentrations of 12 antimicrobials used for treatment of tuberculosis including both first (isoniazid, rifampin, ethambutol) and second line drugs (amikacin, cycloserine, ethionamide, kanamycin, moxifloxacin, ofloxacin, para-aminosalicylic acid, rifabutin, and streptomycin). Pyrazinamide, a first line drug, is not included in the microtiter plate due to its need for acidic test conditions. Advantages of the microtiter system include both ease of set up and faster turn around time (14 days) compared with traditional agar proportion (21 days). In addition, the plate can be set up from inoculum prepared using either broth or solid medium. Since the microtiter plate format is new and since Mtb presents unique safety challenges in the laboratory, this protocol will describe how to safely setup, incubate and read the microtiter plate.

Liquid and vapour-phase antifungal activities of selected essential oils against candida albicans: microscopic observations and chemical characterization of cymbopogon citratus

PubMed Central

2010-01-01

Background Use of essential oils for controlling Candida albicans growth has gained significance due to the resistance acquired by pathogens towards a number of widely-used drugs. The aim of this study was to test the antifungal activity of selected essential oils against Candida albicans in liquid and vapour phase and to determine the chemical composition and mechanism of action of most potent essential oil. Methods Minimum Inhibitory concentration (MIC) of different essential oils in liquid phase, assayed through agar plate dilution, broth dilution & 96-well micro plate dilution method and vapour phase activity evaluated through disc volatilization method. Reduction of C. albicans cells with vapour exposure was estimated by kill time assay. Morphological alteration in treated/untreated C. albicans cells was observed by the Scanning electron microscopy (SEM)/Atomic force microscopy (AFM) and chemical analysis of the strongest antifungal agent/essential oil has been done by GC, GC-MS. Results Lemon grass (Cymbopogon citratus) essential oil exhibited the strongest antifungal effect followed by mentha (Mentha piperita) and eucalyptus (Eucalyptus globulus) essential oil. The MIC of lemon grass essential oil in liquid phase (288 mg/l) was significantly higher than that in the vapour phase (32.7 mg/l) and a 4 h exposure was sufficient to cause 100% loss in viability of C. albicans cells. SEM/AFM of C. albicans cells treated with lemon grass essential oil at MIC level in liquid and vapour phase showed prominent shrinkage and partial degradation, respectively, confirming higher efficacy of vapour phase. GC-MS analysis revealed that lemon grass essential oil was dominated by oxygenated monoterpenes (78.2%); α-citral or geranial (36.2%) and β-citral or neral (26.5%), monoterpene hydrocarbons (7.9%) and sesquiterpene hydrocarbons (3.8%). Conclusion Lemon grass essential oil is highly effective in vapour phase against C. albicans, leading to deleterious morphological changes in cellular structures and cell surface alterations. PMID:21067604

Clonal populations of amniotic cells by dilution and direct plating: evidence for hidden diversity.

PubMed

Wilson, Patricia G; Devkota, Lorna; Payne, Tiffany; Crisp, Laddie; Winter, Allison; Wang, Zhan

2012-01-01

Fetal cells are widely considered a superior cell source for regenerative medicine; fetal cells show higher proliferative capacity and have undergone fewer replicative cycles that could generate spontaneous mutations. Fetal cells in amniotic fluid were among the first normal primary cells to be cultured ex vivo, but the undefined composition of amniotic fluid has hindered advance for regenerative applications. We first developed a highly efficient method to generate clonal populations by dilution of amniocentesis samples in media and direct plating without intervening refrigeration, centrifugation, or exposure of cells to the paracrine effects in mixed cell cultures. More than 40 clonal populations were recovered from 4 amniocentesis samples and representative clones were characterized by flow cytometry, conventional assays for differentiation potential, immunofluorescence imaging, and transcript analysis. The results revealed previously unreported diversity among stromal and epithelial cell types and identified unique cell types that could be lost or undetected in mixed cell populations. The differentiation potential of amniotic cells proved to be uncoupled from expression of definitive cell surface or cytoplasmic markers for stromal and epithelial cells. Evidence for diversity among stromal and epithelial cells in amniotic fluid bears on interpretations applied to molecular and functional tests of amniotic cell populations.

Direct targeting of human plasma for matrix-assisted laser desorption/ionization and analysis of plasma proteins by time of flight-mass spectrometry.

PubMed

Jin, Ya; Manabe, Takashi

2005-07-01

A method to analyze human plasma proteins without fractionation, directly applying a plasma-matrix mixture on the target plate of a matrix-assisted laser desorption/ionization-time of flight-mass spectrometer (MALDI-TOF-MS), has been described. Peaks of ionized plasma proteins could not be detected applying a mixture of an undiluted plasma sample and a matrix solution, but they appeared when the plasma was diluted before mixing with the matrix. Tenfold diluted plasma provided well-resolved protein peaks in the m/z range from 4000 to 30,000. The addition of a simple post-crystallization washing procedure performed on the target plate further improved the quality of mass spectra. We numbered 58 peaks in the range of 4-160 kDa and 32 out of which were assigned to the plasma protein species which have been reported. Especially high sensitivity and resolution were obtained in the region < 30 kDa, where multiple isoforms of apolipoprotein A-I, apolipoprotein A-II, apolipoprotein C-I, apolipoprotein C-II, apolipoprotein C-III, and transthyretin could be assigned. Various post-translational modifications are involved in the isoforms, e.g., proteolytic cleavage, glycosylation and chemical modifications. This method will become complementary with the present electrophoretic techniques, especially for the analysis of low-molecular-mass proteins.

Development and evaluation of an enzyme-linked immunosorbent assay (ELISA) method for the measurement of 2,4-dichlorophenoxyacetic acid in human urine.

PubMed

Chuang, Jane C; Emon, Jeanette M Van; Durnford, Joyce; Thomas, Kent

2005-09-15

An enzyme-linked immunosorbent assay (ELISA) method was developed to quantitatively measure 2,4-dichlorophenoxyacetic acid (2,4-D) in human urine. Samples were diluted (1:5) with phosphate-buffered saline containing 0.05% Tween and 0.02% sodium azide, with analysis by a 96-microwell plate immunoassay format. No clean up was required as dilution step minimized sample interferences. Fifty urine samples were received without identifiers from a subset of pesticide applicators and their spouses in an EPA pesticide exposure study (PES) and analyzed by the ELISA method and a conventional gas chromatography/mass spectrometry (GC/MS) procedure. For the GC/MS analysis, urine samples were extracted with acidic dichloromethane (DCM); methylated by diazomethane and fractionated by a Florisil solid phase extraction (SPE) column prior to GC/MS detection. The percent relative standard deviation (%R.S.D.) of the 96-microwell plate triplicate assays ranged from 1.2 to 22% for the urine samples. Day-to-day variation of the assay results was within +/-20%. Quantitative recoveries (>70%) of 2,4-D were obtained for the spiked urine samples by the ELISA method. Quantitative recoveries (>80%) of 2,4-D were also obtained for these samples by the GC/MS procedure. The overall method precision of these samples was within +/-20% for both the ELISA and GC/MS methods. The estimated quantification limit for 2,4-D in urine was 30ng/mL by ELISA and 0.2ng/mL by GC/MS. A higher quantification limit for the ELISA method is partly due to the requirement of a 1:5 dilution to remove the urine sample matrix effect. The GC/MS method can accommodate a 10:1 concentration factor (10mL of urine converted into 1mL organic solvent for analysis) but requires extraction, methylation and clean up on a solid phase column. The immunoassay and GC/MS data were highly correlated, with a correlation coefficient of 0.94 and a slope of 1.00. Favorable results between the two methods were achieved despite the vast differences in sample preparation. Results indicated that the ELISA method could be used as a high throughput, quantitative monitoring tool for human urine samples to identify individuals with exposure to 2,4-D above the typical background levels.

Detection of microbial concentration in ice-cream using the impedance technique.

PubMed

Grossi, M; Lanzoni, M; Pompei, A; Lazzarini, R; Matteuzzi, D; Riccò, B

2008-06-15

The detection of microbial concentration, essential for safe and high quality food products, is traditionally made with the plate count technique, that is reliable, but also slow and not easily realized in the automatic form, as required for direct use in industrial machines. To this purpose, the method based on impedance measurements represents an attractive alternative since it can produce results in about 10h, instead of the 24-48h needed by standard plate counts and can be easily realized in automatic form. In this paper such a method has been experimentally studied in the case of ice-cream products. In particular, all main ice-cream compositions of real interest have been considered and no nutrient media has been used to dilute the samples. A measurement set-up has been realized using benchtop instruments for impedance measurements on samples whose bacteria concentration was independently measured by means of standard plate counts. The obtained results clearly indicate that impedance measurement represents a feasible and reliable technique to detect total microbial concentration in ice-cream, suitable to be implemented as an embedded system for industrial machines.

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.

Size and Charge Dependence of Ion Transport in Human Nail Plate

PubMed Central

Baswan, Sudhir M.; Li, S. Kevin; LaCount, Terri D.; Kasting, Gerald B.

2016-01-01

The electrical properties of human nail plate are poorly characterized, yet are a key determinate of the potential to treat nail diseases such as onychomycosis using iontophoresis. In order to address this deficiency, molar conductivities of 17 electrolytes comprising 12 ionic species were determined in hydrated human nail plate in vitro. Cation transport numbers across the nail for 11 of these electrolytes were determined by the electromotive force method. Effective ionic mobilities and diffusivities at infinite dilution for all ionic species were determined by regression analysis. The ratios of diffusivities in nail to those in solution were found to correlate inversely with the hydrodynamic radii of the ions according to a power law relationship having an exponent of −1.75 ± 0.27, a substantially steeper size dependence than observed for similar experiments in skin. Effective diffusivities of cations in nail were three-fold higher than those of comparably sized anions. These results reflect the strong size and charge selectivity of the nail plate for ionic conduction and diffusion. The analysis implies that efficient transungual iontophoretic delivery of ionized drugs having radii upwards of 5 Å (approximately MW ≥ 340 Da) will require chemical or mechanical alteration of the nail plate. PMID:26886342

A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method.

PubMed

Rivas, Lucia; Dykes, Gary A; Fegan, Narelle

2007-04-01

Attachment of Shiga toxigenic Escherichia coli (STEC) to surfaces and the formation of biofilms may enhance persistence in a food processing environment and present a risk of contaminating products. Seven strains of STEC and three non-STEC strains were selected to compare two biofilm quantification methods; epifluorescence microscopy on stainless steel (SS) and a microtitre plate assay. The influence of prior growth in planktonic (nutrient broth) and sessile (nutrient agar) culture on biofilm production, as well as expression of surface structures and the possession of antigen 43 (encoded by agn43) on biofilm formation were also investigated. Biofilms were produced in diluted nutrient broth at 25 degrees C for 24 and 48 h. Curli expression was determined using congo red indicator agar, while the presence of agn43 was determined using polymerase chain reaction. No correlation was found between counts for epifluorescence microscopy on SS and the absorbance values obtained with the microtitre plate method for planktonic and sessile grown cultures. Different abilities of individual STEC strains to attach to SS and microtitre plates were found with some strains attaching better to each surface following growth in either planktonic or sessile culture. All O157 STEC strains had low biofilm counts on SS for planktonic and sessile grown cultures; however, one STEC O157:H- strain (EC516) had significantly greater (p<0.05) biofilm production on microtitre plates compared to the other O157 STEC strains. EC516 and other STEC (O174:H21 and O91:H21) strains expressing curli fimbriae were found to produce significantly greater (p<0.05) biofilms on microtitre plates compared to the non-curli expressing strains. No relationship was found between the production of type-I fimbriae, motility, agn43 and bacterial physicochemical properties (previously determined) and biofilm formation on SS or microtitre plates. Variations between the two biofilm determination methods may suggest that the biofilm production on microtitre plates may not be appropriate to represent other surfaces such as SS and that caution should be taken when selecting a method to quantify biofilm production on a surface.

In Vitro Activity of Voriconazole against Prototheca wickerhamii: Comparative Evaluation of Sensititre and NCCLS M27-A2 Methods of Detection

PubMed Central

Linares, Maria José; Solís, Francisco; Casal, Manuel

2005-01-01

A total of 104 Prototheca wickerhamii isolates and two control strains were tested for susceptibility to voriconazole using the Sensititre YeastOne colorimetric antifungal plate and NCCLS reference method. Voriconazole was highly active against all isolates, with an MIC at which 90% of isolates were inhibited of ≤0.5 μg/ml. Comparison of MICs obtained with the Sensititre product and the NCCLS method demonstrated agreement (100% ± 2 log2 dilutions) between the two methods. Voriconazole may offer an option for the treatment of Prototheca sp. infections, and its efficacy should be established through clinical experience. PMID:15872301

Finite-Difference Solutions for Compressible Laminar Boundary-Layer Flows of a Dusty Gas over a Semi-Infinite Flat Plate.

DTIC Science & Technology

1986-08-01

AD-A174 952 FINITE - DIFFERENCE SOLUTIONS FOR CONPRESSIBLE LANINAR 1/2 BOUNDARY-LAYER FLOUS (U) TORONTO UNIV DOWNSVIEW (ONTARIO) INST FOR AEROSPACE...dilute dusty gas over a semi-infinite flat plate. Details are given of the impliit finite , difference schemes as well as the boundary conditions... FINITE - DIFFERENCE SOLUTIONS FOR COMPRESSIBLE LAMINAR BOUNDARY-LAYER FLOWS OF A DUSTY GAS OVER A SEMI-INFINITE FLAT PLATE by B. Y. Wang and I. I

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.

The Antibacterial Activity of Honey on Helicobacter Pylori

PubMed Central

Nzeako, Basil C; Al-Namaani, Faiza

2006-01-01

Objective: This project aimed to assess the antibacterial potential of various brands of honey sold in Muscat area on some isolates of H. pylori and to determine if there is any synergy between honey and amoxycillin or clarithromycin used in the treatment of H. pylori gastritis and duodenal ulcer. Methods: Eight samples of commercial honey were used in the experiment after they were checked for purity by sub-culturing on blood agar and incubating for 48 hours at 37°c. Honey samples showing gross contamination were discarded. Purified culture isolates of H. pylori from our laboratory stock cultures were swabbed on chocolate plate using 1x 104 cfu/ml. One hundred microlitres (100μl) of various honey samples were placed on each plate which was subsequently incubated microaerophilically at 37ºc for 3 days. The presence or absence of growth inhibition zones on each plate was noted and an average zone size of each honey was taken. Honey samples with high zone sizes were further diluted from 1:2–1:8 to find the end-points of their growth inhibition concentrations and the experiment was repeated in triplicates. The synergistic effect between honey, amoxycillin and clarithromycin was done in triplicates by placing honey at various distances between each antibiotic after swabbing chocolate agar with 1x 104 cfu/ml of H. pylori. The plates were incubated as before. Results: All honey samples produced growth inhibition zones with H. pylori no at dilution of honey but had different zone sizes at 1:2–1:8 dilutions. Black Forest honey had the highest antibacterial activity followed by Langnese honey. None of the honeys had a synergistic effect with either clarithromycin or amoxycillin. Conclusion: We conclude that, in vitro, some honey brands possess antibacterial activity against H. pylori and that no synergy or antagonism was observed between honey and clarithromycin or honey and amoxicillin using H. pylori as a test organism. Though no synergy or antagonism was observed between honey, amoxicillin or clarithromycin, it has been suggested that the use of honey with triple therapy regimen may help shorten the time required to eliminate H. pylori from stomach lining of patients with gastritis or duodenal ulcer caused by H. pylori infection PMID:21748138

Clonal Populations of Amniotic Cells by Dilution and Direct Plating: Evidence for Hidden Diversity

PubMed Central

Wilson, Patricia G.; Devkota, Lorna; Payne, Tiffany; Crisp, Laddie; Winter, Allison; Wang, Zhan

2012-01-01

Fetal cells are widely considered a superior cell source for regenerative medicine; fetal cells show higher proliferative capacity and have undergone fewer replicative cycles that could generate spontaneous mutations. Fetal cells in amniotic fluid were among the first normal primary cells to be cultured ex vivo, but the undefined composition of amniotic fluid has hindered advance for regenerative applications. We first developed a highly efficient method to generate clonal populations by dilution of amniocentesis samples in media and direct plating without intervening refrigeration, centrifugation, or exposure of cells to the paracrine effects in mixed cell cultures. More than 40 clonal populations were recovered from 4 amniocentesis samples and representative clones were characterized by flow cytometry, conventional assays for differentiation potential, immunofluorescence imaging, and transcript analysis. The results revealed previously unreported diversity among stromal and epithelial cell types and identified unique cell types that could be lost or undetected in mixed cell populations. The differentiation potential of amniotic cells proved to be uncoupled from expression of definitive cell surface or cytoplasmic markers for stromal and epithelial cells. Evidence for diversity among stromal and epithelial cells in amniotic fluid bears on interpretations applied to molecular and functional tests of amniotic cell populations. PMID:23024659

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

Size and Charge Dependence of Ion Transport in Human Nail Plate.

PubMed

Baswan, Sudhir M; Li, S Kevin; LaCount, Terri D; Kasting, Gerald B

2016-03-01

The electrical properties of human nail plate are poorly characterized yet are a key determinate of the potential to treat nail diseases, such as onychomycosis, using iontophoresis. To address this deficiency, molar conductivities of 17 electrolytes comprising 12 ionic species were determined in hydrated human nail plate in vitro. Cation transport numbers across the nail for 11 of these electrolytes were determined by the electromotive force method. Effective ionic mobilities and diffusivities at infinite dilution for all ionic species were determined by regression analysis. The ratios of diffusivities in nail to those in solution were found to correlate inversely with the hydrodynamic radii of the ions according to a power law relationship having an exponent of -1.75 ± 0.27, a substantially steeper size dependence than observed for similar experiments in skin. Effective diffusivities of cations in nail were 3-fold higher than those of comparably sized anions. These results reflect the strong size and charge selectivity of the nail plate for ionic conduction and diffusion. The analysis implies that efficient transungual iontophoretic delivery of ionized drugs having radii upward of 5 Å (molecular weight, ca. ≥ 340 Da) will require chemical or mechanical alteration of the nail plate. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Validation of FMTV Modular VHP/mVHP System and Fumigation Decontamination Process in a C-141B Starlifter Aircraft

DTIC Science & Technology

2007-08-01

each location was aseptically transferred to 5 mL Tryptic Soy Broth (TSB) and incubated at 55 ’C. Coupons were observed the following day. If...Samples were then serially diluted in buffered peptone water and pour plated (1 mL per plate) using Tryptic Soy Agar (TSA). Plates were gently swirled in...1260 area. aft surface inside of hydraulic oil box 19 39 Starboard - 750. 59 On V2 79 On platform, 99 Aft, overhead in on oxygen box overhead, forward

A comparison of the Sensititre® MYCOTB panel and the agar proportion method for the susceptibility testing of Mycobacterium tuberculosis.

PubMed

Abuali, M M; Katariwala, R; LaBombardi, V J

2012-05-01

The agar proportion method (APM) for determining Mycobacterium tuberculosis susceptibilities is a qualitative method that requires 21 days in order to produce the results. The Sensititre method allows for a quantitative assessment. Our objective was to compare the accuracy, time to results, and ease of use of the Sensititre method to the APM. 7H10 plates in the APM and 96-well microtiter dry MYCOTB panels containing 12 antibiotics at full dilution ranges in the Sensititre method were inoculated with M. tuberculosis and read for colony growth. Thirty-seven clinical isolates were tested using both methods and 26 challenge strains of blinded susceptibilities were tested using the Sensititre method only. The Sensititre method displayed 99.3% concordance with the APM. The APM provided reliable results on day 21, whereas the Sensititre method displayed consistent results by day 10. The Sensititre method provides a more rapid, quantitative, and efficient method of testing both first- and second-line drugs when compared to the gold standard. It will give clinicians a sense of the degree of susceptibility, thus, guiding the therapeutic decision-making process. Furthermore, the microwell plate format without the need for instrumentation will allow its use in resource-poor settings.

Biodiversity within hot spring microbial mat communities: molecular monitoring of enrichment cultures

NASA Technical Reports Server (NTRS)

Ward, D. M.; Santegoeds, C. M.; Nold, S. C.; Ramsing, N. B.; Ferris, M. J.; Bateson, M. M.

1997-01-01

We have begun to examine the basis for incongruence between hot spring microbial mat populations detected by cultivation or by 16S rRNA methods. We used denaturing gradient gel electrophoresis (DGGE) to monitor enrichments and isolates plated therefrom. At near extincting inoculum dilutions we observed Chloroflexus-like and cyanobacterial populations whose 16S rRNA sequences have been detected in the 'New Pit' Spring Chloroflexus mat and the Octopus Spring cyanobacterial mat. Cyanobacterial populations enriched from 44 to 54 degrees C and 56 to 63 degrees C samples at near habitat temperatures were similar to those previously detected in mat samples of comparable temperatures. However, a lower temperature enrichment from the higher temperature sample selected for the populations found in the lower temperature sample. Three Thermus populations detected by both DGGE and isolation exemplify even more how enrichment may bias our view of community structure. The most abundant population was adapted to the habitat temperature (50 degrees C), while populations adapted to 65 degrees C and 70 degrees C were 10(2)- and 10(4)-fold less abundant, respectively. However, enrichment at 70 degrees C favored the least abundant strain. Inoculum dilution and incubation at the habitat temperature favored the more numerically relevant populations. We enriched many other aerobic chemoorganotrophic populations at various inoculum dilutions and substrate concentrations, most of whose 16S rRNA sequences have not been detected in mats. A common feature of numerically relevant cyanobacterial, Chloroflexus-like and aerobic chemorganotrophic populations, is that they grow poorly and resist cultivation on solidified medium, suggesting plating bias, and that the medium composition and incubation conditions may not reflect the natural microenvironments these populations inhabit.

Rapid enumeration of viable bacteria by image analysis

NASA Technical Reports Server (NTRS)

Singh, A.; Pyle, B. H.; McFeters, G. A.

1989-01-01

A direct viable counting method for enumerating viable bacteria was modified and made compatible with image analysis. A comparison was made between viable cell counts determined by the spread plate method and direct viable counts obtained using epifluorescence microscopy either manually or by automatic image analysis. Cultures of Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Yersinia enterocolitica and Pseudomonas aeruginosa were incubated at 35 degrees C in a dilute nutrient medium containing nalidixic acid. Filtered samples were stained for epifluorescence microscopy and analysed manually as well as by image analysis. Cells enlarged after incubation were considered viable. The viable cell counts determined using image analysis were higher than those obtained by either the direct manual count of viable cells or spread plate methods. The volume of sample filtered or the number of cells in the original sample did not influence the efficiency of the method. However, the optimal concentration of nalidixic acid (2.5-20 micrograms ml-1) and length of incubation (4-8 h) varied with the culture tested. The results of this study showed that under optimal conditions, the modification of the direct viable count method in combination with image analysis microscopy provided an efficient and quantitative technique for counting viable bacteria in a short time.

False positives in Biolog EcoPlates™ and MT2 MicroPlates™ caused by calcium.

PubMed

Pierce, Melissa L; Ward, J Evan; Dobbs, Fred C

2014-02-01

Biolog MicroPlates(TM) (e.g. EcoPlate(TM), MT2 MicroPlate(TM), GN MicroPlate(TM)) are useful tools for characterizing microbial communities, providing community-level physiological profiles to terrestrial and aquatic ecologists. The more recently designed Biolog EcoPlates have been used frequently in aquatic ecology with success. This study, however, reveals one major problem when using EcoPlates to evaluate samples within an estuarine or seawater matrix. At concentrations greater than 100 parts per million, the cation calcium begins to interfere with the microplate chemistry, causing false positive readings. Experiments, in which multiple treatments of natural and artificial seawater were tested, as well as calcium-addition experiments, demonstrate that calcium inhibits complete dissolution of the minimal growth medium in wells. Future studies involving Biolog EcoPlates and MicroPlates should take this effect into account, and the dilution of samples is strongly recommended to diminish the "calcium effect." Copyright © 2013 Elsevier B.V. All rights reserved.

Concepts for a low-vibration and cryogen-free tabletop dilution refrigerator

NASA Astrophysics Data System (ADS)

Uhlig, Kurt

2017-10-01

The purpose of this article is to describe several concepts of how to cool a modern tabletop dilution refrigerator (DR) with a cryogen-free pulse tube cryocooler (PTC). Tabletop DRs have come more and more into the focus of scientists, recently, because they offer easy access to the mixing chamber mounting plate from all directions and because of their very short cooldown times. However, these milli-Kelvin coolers are precooled with LHe which makes their handling inconvenient and often expensive. In the paper it is explained how a cryocooler can be directly coupled to a DR unit making the use of LHe superfluous. Furthermore, concepts are discussed where a tabletop DR is cooled by a remote PTC; PTC and DR are mounted in separate vacuum containers which are connected by a stainless steel bellows tube. This kind of apparatus would offer an extremely low level of vibration at the mixing chamber mounting plate.

Microstructures and mechanical properties of bonding layers between low carbon steel and alloy 625 processed by gas tungsten arc welding

NASA Astrophysics Data System (ADS)

Lou, Shuai; Lee, Seul Bi; Nam, Dae-Geun; Choi, Yoon Suk

2017-11-01

A filler metal wire, Alloy 625, was cladded on a plate of a low carbon streel, SS400, by gas tungsten arc welding, and the morphology of the weld bead and resulting dilution ratio were investigated under different welding parameter values (the input current, weld speed and wire feed speed). The wire feed speed was found to be most influential in controlling the dilution ratio of the weld bead, and seemed to limit the influence of other welding parameters. Two extreme welding conditions (with the minimum and maximum dilution ratios) were identified, and the corresponding microstructures, hardness and tensile properties near the bond line were compared between the two cases. The weld bead with the minimum dilution ratio showed superior hardness and tensile properties, while the formation lath martensite (due to relatively fast cooling) affected mechanical properties in the heat affected zone of the base metal with the maximum dilution ratio.

A method for detecting fungal contaminants in wall cavities.

PubMed

Spurgeon, Joe C

2003-01-01

This article describes a practical method for detecting the presence of both fungal spores and culturable fungi in wall cavities. Culturable fungi were collected in 25 mm cassettes containing 0.8 microm mixed cellulose ester filters using aggressive sampling conditions. Both culturable fungi and fungal spores were collected in modified slotted-disk cassettes. The sample volume was 4 L. The filters were examined microscopically and dilution plated onto multiple culture media. Collecting airborne samples in filter cassettes was an effective method for assessing wall cavities for fungal contaminants, especially because this method allowed the sample to be analyzed by both microscopy and culture media. Assessment criteria were developed that allowed the sample results to be used to classify wall cavities as either uncontaminated or contaminated. As a criterion, wall cavities with concentrations of culturable fungi below the limit of detection (LOD) were classified as uncontaminated, whereas those cavities with detectable concentrations of culturable fungi were classified as contaminated. A total of 150 wall cavities was sampled as part of a field project. The concentrations of culturable fungi were below the LOD in 34% of the samples, whereas Aspergillus and/or Penicillium were the only fungal genera detected in 69% of the samples in which culturable fungi were detected. Spore counting resulted in the detection of Stachybotrys-like spores in 25% of the samples that were analyzed, whereas Stachybotrys chartarum colonies were only detected on 2% of malt extract agar plates and on 6% of corn meal agar plates.

Dial-A-Decon Solution Chemistry GAP Testing

DTIC Science & Technology

2012-04-01

34 The tubes were serially diluted using Buttcrfield’s buffer solution and plated in triplicate on Tryptic Soy Agar. Plates were enumerated the...of 200 uL HD to 10 mL of the surfactant solution. The energy to create the oil in water (O/W) emulsions was provided by magnetic stirring. Solutions...emulsify a mixture of water and oil such as HD, one or more emulsifiers are required. Each surfactant system can be characterized by an HLB value

Comparative evaluation of antimicrobial efficacy of sodium hypochlorite, MTAD, and Tetraclean against Enterococcus faecalis biofilm.

PubMed

Giardino, Luciano; Ambu, Emanule; Savoldi, Enrico; Rimondini, Roberto; Cassanelli, Clara; Debbia, Eugenio A

2007-07-01

The aim of this study was to compare the antimicrobial efficacy of 5.25% NaOCl, BioPure MTAD (Dentsply Tulsa Dental, Johnson City, TN), and Tetraclean (Ogna Laboratori Farmaceutici, Milano, Italy) against Enterococcus faecalis biofilm generated on cellulose nitrate membrane filters. After incubation, the membrane filters were transferred into tubes containing 5 mL of the selected antimicrobial solution test agent or NaCl 0.9% (positive control) and incubated for 5, 30, and 60 minutes at 20 degrees C. After each period of time, the test agents were vortexed for 60 seconds to resuspend the microorganisms. Ten-fold serial dilutions were generated in reduced transport fluid. Each dilution was plated onto a brain heart infusion plates. The plates were then incubated for 48 hours in an aerobic atmosphere at 37 degrees C and colony-forming units per membrane was calculated. Statistical analysis showed that only 5.25% NaOCl can disgregate and remove the biofilm at every time; however, treatment with Tetraclean caused a high degree of biofilm disgregation in every considered time intervals as compared with MTAD (T5 p < 0.05, T30 p < 0.01, and T60 p < 0.001).

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.

Assessing the growth and recovery of Salmonella Enteritidis SE86 after sodium dichloroisocyanurate exposure

PubMed Central

Ferreira, Fernanda Stoduto; Horvath, Mariana Bandeira; Tondo, Eduardo Cesar

2013-01-01

The objective of the present study was to assess the growth and the recovery of Salmonella (S.) Enteritidis SE86 in different diluents, culture media and using different plating methods after the exposure to 200 mg/kg sodium dichloroisocyanurate (NaDCC). Before and after NaDCC exposure, SE86 was cultured at 30 °C and 7 °C in the following diluents: Peptone water (P), Saline solution (SaS), Peptone water+Saline solution (P+SaS), Peptone water+Tween 80+Lecithin+Sodium thiosulfate (P+N) and Saline solution+Tween 80+Lecithin+Sodium thiosulfate (SaS+N). The SaS diluent was chosen because it was able to maintain cells viable without growth and was further used for plating SE86 on non selective medium (Tryptic Soy Agar-TSA) and on selective media (Mannitol Lysine Crystal Violet Brilliant Green Agar-MLCB; Brilliant Green Agar-BGA; Salmonella Shigella Agar-SS and Xylose Lysine Dextrose–XLD). The Thin Agar Layer method (TAL) i.e., selective media overlayed with non selective TSA was also evaluated. Results indicated that SE86 not exposed to NaDCC was able to grow in P, P+N, SaS+N and P+SaS, but not in SaS, that was able to maintain cells viable. SE86 exposed to NaDCC demonstrated similar counts after dilution in SaS and the plating on non selective TSA, selective media MLCB, BGA, SS and XLD and on TAL media. SE86, S. Typhimurium and S. Bredeney, exposed or not exposed to NaDCC, showed no significant differences in counts on TSA, XLD and XLD overlayed with TSA, suggesting that all those media may be used to quantify NaDCC-exposed Salmonella by plating method. PMID:24516446

Delivery of Formulated Industrial Enzymes with Acoustic Technology.

PubMed

Hwang, Jennifer Dorcas; Ortiz-Maldonado, Mariliz; Paramonov, Sergey

2016-02-01

Industrial enzymes are instrumental in many applications, including carbohydrate processing, fabric and household care, biofuels, food, and animal nutrition, among others. Enzymes have to be active and stable not only in harsh application conditions, but also during shipment and storage. In protein stability studies, formulated concentrated enzyme solutions are frequently diluted gravimetrically prior to enzyme activity measurements, making it challenging to move toward more high-throughput techniques using conventional robotic equipment. Current assay methods pose difficulties when measuring highly concentrated proteins. For example, plastic pipette tips can introduce error because proteins adsorb to the tip surface, despite the presence of detergents, decreasing precision and overall efficiency of protein activity assays. Acoustic liquid handling technology, frequently used for various dilute small-molecule assays, may overcome such problems. Originally shown to effectively deliver dilute solutions of small molecules, this technology is used here as an effective alternative to the aforementioned challenge with viscous concentrated protein solutions. Because the acoustic liquid handler transfers nanoliter quantities of liquids without using pipette tips and without sample loss, it rapidly and uniformly prepares assay plates for enzyme activity measurements within minutes. This increased efficiency transforms the nature of enzyme stability studies toward high precision and throughput. © 2015 Society for Laboratory Automation and Screening.

Development of a rapid method to quantify Salmonella Typhimurium using a combination of MPN with qPCR and a shortened time incubation.

PubMed

Kim, Sun Ae; Park, Si Hong; Lee, Sang In; Ricke, Steven C

2017-08-01

A novel method was developed for the specific quantification of S. Typhimurium using a most-probable-number (MPN) combined with qPCR and a shortened incubation time (MPN-qPCR-SIT). For S. Typhimurium enumeration, dilutions of samples were transferred into three wells on a microtiter plate and the plate was incubated for 4 h. The S. Typhimurium presence in the wells was identified using a qPCR and populations were determined based on an MPN calculation. The R 2 between the MPN-qPCR-SIT and conventional MPN exhibited a high level of correlation (0.9335-0.9752), suggesting that the MPN-qPCR-SIT offers a reliable alternative method for S. Typhimurium quantification. Although plating and qPCR were limited in their ability to detect low levels of S. Typhimurium (e.g. 0.18 log MPN/ml), these levels could be successfully detected with the MPN-qPCR-SIT. Chicken breast samples inoculated with S. Typhimurium were incubated at 0, 4, and 24 h and incubated samples were subjected to microbiome analysis. Levels of Salmonella and Enterobacteriaceae increased significantly with incubation time. The obvious benefits of the MPN-qPCR-SIT are: 1) a further confirmation step is not required, 2) the detection limit is as low as conventional MPN, but 3) is more rapid, requiring approximately 7 h to simultaneously complete quantification. Copyright © 2017 Elsevier Ltd. All rights reserved.

Porphyromonas gingivalis can invade periodontal ligament stem cells.

PubMed

Pan, Chunling; Liu, Junchao; Wang, Hongyan; Song, Jia; Tan, Lisi; Zhao, Haijiao

2017-02-17

Porphyromonas gingivalis is strongly associated with the development, progression, severity and recurrence of periodontitis. Periodontal ligament stem cells (PDLSCs) play an important role in the maintenance of periodontal tissue self-renewal and repair. The purpose of this study was to investigate the ability of P. gingivalis to infect PDLSCs using an in vitro monolayer model. We separated and cultured primary PDLSCs using the tissue block with limiting dilution method. The efficiency of P. gingivalis (ATCC 33277) infection of PDLSCs was measured using agar plate culture and quantitative polymerase chain reaction (q-PCR) methods. PDLSCs infected with P. gingivalis were also observed by transmission electron microscopy. We assessed stem cell properties including cell morphology, clone formation, growth activity, cell surface antigens and multiple differentiation capacity. The infection rates of P. gingivalis in PDLSC at MOIs of 50, 100, 200, and 500 were 5.83%, 8.12%, 7.77% and 7.53% according to the agar plate culture method. By q-PCR, the efficiencies of P. gingivalis infection of PDLSCs at MOIs of 50, 100, 200, and 500 were 6.74%, 10.56%, 10.36% and 9.78%, respectively. Overall, the infection efficiency based on q-PCR was higher than that according to agar plate culture. Using transmission electron microscopy, we verified that P. gingivalis (ATCC 33277) could infect and invade PDLSCs after 2 h of incubation, and endocytic vacuoles were not found surrounding the internalized bacteria. In conclusion, our data demonstrate that P. gingivalis can invade PDLSCs.

New procedure to reduce the time and cost of broncho-pulmonary specimen management using the Previ Isola® automated inoculation system.

PubMed

Nebbad-Lechani, Biba; Emirian, Aurélie; Maillebuau, Fabienne; Mahjoub, Nadia; Fihman, Vincent; Legrand, Patrick; Decousser, Jean-Winoc

2013-12-01

The microbiological diagnosis of respiratory tract infections requires serial manual dilutions of the clinical specimen before agar plate inoculation, disrupting the workflow in bacteriology clinical laboratories. Automated plating instrument systems have been designed to increase the speed, reproducibility and safety of this inoculating step; nevertheless, data concerning respiratory specimens are lacking. We tested a specific procedure that uses the Previ Isola® (bioMérieux, Craponne, France) to inoculate with broncho-pulmonary specimens (BPS). A total of 350 BPS from a university-affiliated hospital were managed in parallel using the manual reference and the automated methods (expectoration: 75; broncho-alveolar lavage: 68; tracheal aspiration: 17; protected distal sample: 190). A specific enumeration reading grid, a pre-liquefaction step and a fluidity test, performed before the inoculation, were designed for the automated method. The qualitative (i.e., the number of specimens yielding a bacterial count greater than the clinical threshold) and quantitative (i.e., the discrepancy within a 0.5 log value) concordances were 100% and 98.2%, respectively. The slimmest subgroup of expectorations could not be managed by the automated method (8%, 6/75). The technical time and cost savings (i.e., number of consumed plates) reached 50%. Additional studies are required for specific populations, such as cystic fibrosis specimens and associated bacterial variants. An automated decapper should be implemented to increase the biosafety of the process. The PREVI Isola® adapted procedure is a time- and cost-saving method for broncho-pulmonary specimen processing. © 2013.

Silver-halide sensitized gelatin (SHSG) processing method for pulse holograms recorded on VRP plates

NASA Astrophysics Data System (ADS)

Evstigneeva, Maria K.; Drozdova, Olga V.; Mikhailov, Viktor N.

2002-06-01

One of the most important area of holograph applications is display holography. In case of pulse recording the requirement for vibration stability is easier than compared to CW exposure. At the same time it is widely known that the behavior of sliver-halide holographic materials strongly depends on the exposure duration. In particular the exposure sensitivity drastically decreases under nanosecond pulse duration. One of the effective ways of the diffraction efficiency improvement is SHSG processing method. This processing scheme is based on high modulation of refractive index due to microvoids appearance inside emulsion layer. It should be mentioned that the SHSG method was used earlier only in the cases when the holograms were recorded by use of CW lasers. This work is devoted to the investigation of SHSG method for pulse hologram recording on VRP plates. We used a pulsed YLF:Nd laser with pulse duration of 25 nanoseconds and wavelength of 527 nm. Both transmission and reflection holograms were recorded. The different kinds of bleaching as well as developing solutions were investigated. Our final processing scheme includes the following stages: 1) development in non-tanning solution, 2) rehalogenating bleach, 3) intermediate alcohol drying, 4) uniform second exposure, 5) second development in diluted developer, 6) reverse bleaching, 7) fixing and 8) gradient drying in isopropyl alcohol. Diffraction efficiency of transmission holograms was of about 60 percent and reflection mirror holograms was of about 45 percent. Thus we have demonstrated the SHSG processing scheme for producing effective holograms on VRP plates under pulse exposure.

Comparison of 7 culture methods for Salmonella serovar Enteritidis and Salmonella serovar Typhimurium isolation in poultry feces.

PubMed

Rodríguez, Francisco I; Procura, Francisco; Bueno, Dante J

2018-06-26

The present work compared 7 different culture methods and 3 selective-differential plating media for Salmonella ser. Enteritidis (SE) and S. ser. Typhimurium (ST) isolation using artificially contaminated poultry feces. The sensitivity (Se) and accuracy (AC) values increased when Modified Semisolid Rappaport Vassiliadis (MSRV) methods were used in place of the Tetrathionate (TT) or Tetrathionate Hajna broth (TTH) method in the enrichment step. However, there was no significant difference between the pre-enrichment incubation at 4 to 6 and 18 to 24 h for MSRV5 and MSRV24 methods, respectively. All Salmonella strains were recovered in the lowest dilutions tested for MSRV24 and 3 out of 4 for MSRV5 methods (2 to 10 cfu/25 g). The TT and TTH methods showed a detection limit between 2.2 × 101 and 1.0 × 106 cfu/25 g of fecal sample. The agreement was variable between the methods. However, there was a very good agreement between the MSRV5 and MSRV24 methods, and between tetrathionate direct (TTD, no pre-enrichment media used) and buffered peptone water 18 to 24 h Tetrathionate broth combination (TT24 method) for Salmonella strains. The 3 selective-differential plating media showed an agreement between fair and excellent. They performed a high Se and AC in the MSRV methods for Salmonella strains. There was a significant difference between center and periphery for MSRV methods, and there was a fair agreement between them for all strains. The MSRV methods are better than TT/TTH methods for the isolation of different strains of SE and ST in poultry fecal samples. The MSRV5 method can be used to reduce the time for the detection of SE and ST in these samples. Furthermore, a loopful of the periphery of the growth should be streaked onto differential-selective plating media, even in the absence of halo, to decrease the number of false negative results.

Improved method for mutagenicity testing of gaseous compounds by using a gas sampling bag.

PubMed

Araki, A; Noguchi, T; Kato, F; Matsushima, T

1994-05-01

A simple and safety gas exposure method was developed using a gas sampling bag as an exposure vessel and a preparation vessel of diluted gas. The gas exposure conditions such as amount of S9 in the plate, volume of gas for the plate, amount of top agar, exposure period and exposure temperature were examined by mutagenicity testing of 1,3-butadiene using the gas sampling bag. Mutagenicity tests of 14 compounds and 1,3-butadiene on S. typhimurium TA98, TA100, TA1535 and TA1537, and E. coli WP2 uvrA were also examined by the developed gas exposure method. 1,3-Butadiene, propyne (methyl acetylene), monochlorodifluoromethane, ethylchloride, diborane and silane were mutagenic. 1-Butene, 2-butene, 2-methylpropene, methyl vinyl ether, trichlorofluoromethane, dichlorodifluoromethane, 1,2-dichloro-1,1,2,2-tetrafluoroethane, 1,1-difluoroethane and phosphine were not mutagenic on S. typhimurium TA98, TA100, TA1535 and TA1537, and E. coli WP2 uvrA with or without metabolic activation. These results were compatible with a previous report, and this developed method has the advantage that it can be tested easily and safely for combustible and self-combustible substances such as 1,3-butadiene and silane.

Optical and other property changes of M-50 bearing steel surfaces for different lubricants and additive prior to scuffing

NASA Technical Reports Server (NTRS)

Lauer, J. L.; Marxer, N.

1984-01-01

An ester lubricant base oil containing one or more standard additives to protect against wear, corrosion, and oxidation was used in an experimental ball/plate elastohydrodynamic contact under load and speed conditions such as to induce scuffing failure in short times. Both the ball and the plate were of identically treated M-50 steel. After various periods of operating time the wear track on the plate was examined with an interference microscope of plus or minus 30 A depth resolution and sometimes also with a scanning ellipsometer and an Auger spectrometer. The optically deduced surface profiles varied with wavelength, indicating the presence of surface coatings, which were confirmed by the other instruments. As scuffing was approached, a thin (approximately A) oxide layer and a carbide layer formed in the wear track in particular when tricresylphosphate antiwear additive was present in the lubricant. The rates of the formation of these layers and their reactivity toward dilute alcholic HCl depended strongly on the lubricant and additives. Based on these results suggestions for improved formulations and a test method for bearing reliability could be proposed.

Study of gas tungsten arc welding procedures for tantalum alloy T-111 (Ta-8 W-2Hf) plate

NASA Technical Reports Server (NTRS)

Gold, R. E.; Kesterson, R. L.

1973-01-01

Methods of eliminating or reducing underbread cracking in multipass GTA welds in thick T-111 plate were studied. Single V butt welds prepared using experimental filler metal compositions and standard weld procedures resulted in only moderate success in reducing underbread cracking. Subsequent procedural changes incorporating manual welding, slower weld speeds, and three or fewer fill passes resulted in crack-free single V welds only when the filler metal was free of hafnium. The double V joint design with successive fill passes on opposite sides of the joint produced excellent welds. The quality of each weld was determined metallographically since the cracking, when present, was very slight and undetectable using standard NDT techniques. Tensile and bend tests were performed on selected weldments. The inherent filler metal strength and the joint geometry determined the strength of the weldment. Hardness and electron beam microprobe traverses were made on selected specimens with the result that significant filler metal-base metal dilution as well as hafnium segregation was detected. A tentative explanation of T-111 plate underbread cracking is presented based on the intrinsic effects of hafnium in the weldment.

Confirmation of Two Undescribed Fungal Species from Dokdo of Korea Based on Current Classification System Using Multi Loci

PubMed Central

Lee, Hye Won; Nguyen, Thi Thuong Thuong; Mun, Hye Yeon; Lee, Haengsub; Kim, Changmu

2015-01-01

Using dilution plating method, 47 fungal isolates were obtained from a soil sample collected from Dokdo in the East Sea of Korea in 2013. In this study, two fungal isolates, EML-MFS30-1 and EML-DDSF4, were confirmed as undescribed species, Metarhizium guizhouense and Mortierella oligospora in Korea based on current classification system using multi loci including rDNA internal transcribed spacer, large subunit, small subunit, and β-tubulin (BTUB) genes. Herein, detailed morphological descriptions on characters of the undescribed fungal species as well as their molecular phylogenetic status are provided with comparisons to related species. PMID:26839498

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.

Assessment of antinociceptive, antipyretic and antimicrobial activity of Piper cubeba L. essential oil in animal models.

PubMed

Mothana, Ramzi; Alsaid, Mansour; Khaled, Jamal M; Alharbi, Naiyf S; Alatar, Abdulrahman; Raish, Mohammad; Al-Yahya, Mohammed; Rafatullah, Syed; Parvez, Mohammad Khalid; Ahamad, Syed Rizwan

2016-03-01

This study was designed to investigate the possible antiniciceptive, antipyretic and antimicrobial activities of the essential oil obtained from the fruits of Piper Cubeba (L.). To assess the antinociceptive and antipyretic activities, three doses (150, 300 and 600 mg/kg, i.p.) were tested in acetic acid-induced abdominal writhing, tail flick reaction and hot-plate and Brewer's yeast-induced hyperpyrexia test models in animals. Moreover, the antimicrobial activity was examined using agar diffusion method and broth micro-dilution assay for minimum inhibitory concentrations (MIC). The Piper Cubeba essential oil (PCEO) showed a marked antinociception (17, 30 and 54%) and an increase in reaction time in mice in the flick tailed and hot-plate tests. The brewer's yeast induced hyperpyrexia was decreased in a dose dependent manner. PCEO also exhibited a strong antimicrobial potential. These findings confirm the traditional analgesic indications of P. cubeba oil and provide persuasive evidence and support its use in Arab traditional medicine.

Mycoflora and mycotoxin-producing fungi of air-dust particles from Egypt.

PubMed

Abdel-Hafez, S I; Shoreit, A A; Abdel-Hafez, A I; el Maghraby, O M

1986-01-01

Using the dilution-plate method, 27 genera and 64 species were collected from 20 air-dust samples on glucose - (24 genera and 57 species) and cellulose - (21 genera and 45 species) Czapek's agar at 28 degrees C. There are basic similarities between the mycoflora of air-dust on the two media and the most prevalent species were Aspergillus niger, A. flavus, A. ochraceus, A. terreus, A. versicolor, Penicillium chrysogenum, P. funiculosum, Alternaria alternata, Cladosporium herbarum, Fusarium oxysporum, Rhizopus stolonifer and Trichoderma viride. Chaetomium globosum, Stachybotrys chartarum, Humicola grisea and Arthrobotrys oligospora were common only on cellulose agar plates. Extracts of mycelium from 25 isolates were tested with brine schrimp (Artemia salina); of these 23 displayed varying degrees of toxicity. Thin layer chromatographic analysis of 12 isolates of Aspergillus flavus revealed that 4 strains were producing detectable aflatoxin. Zearalenone production was noted for 3 out of 5 strains of Fusarium oxysporum and 2 out of 5 strains of F. solani.

Suppression of dilution in Ni-Cr-Si-B alloy cladding layer by controlling diode laser beam profile

NASA Astrophysics Data System (ADS)

Tanigawa, Daichi; Funada, Yoshinori; Abe, Nobuyuki; Tsukamoto, Masahiro; Hayashi, Yoshihiko; Yamazaki, Hiroyuki; Tatsumi, Yoshihiro; Yoneyama, Mikio

2018-02-01

A Ni-Cr-Si-B alloy layer was produced on a type 304 stainless steel plate by laser cladding. In order to produce cladding layer with smooth surface and low dilution, influence of laser beam profile on cladding layer was investigated. A laser beam with a constant spatial intensity at the focus spot was used to suppress droplet formation during the cladding layer formation. This line spot, formed with a focussing unit designed by our group, suppressed droplet generation. The layer formed using this line spot with a constant spatial intensity had a much smoother surface compared to a layer formed using a line spot with a Gaussian-like beam. In addition, the dilution of the former layer was much smaller. These results indicated that a line spot with a constant spatial intensity was more effective in producing a cladding layer with smooth surface and low dilution because it suppressed droplet generation.

Rathayibacter iranicus isolated from asymptomatic wheat seeds in Turkey

USDA-ARS?s Scientific Manuscript database

Asymptomatic wheat seeds collected from 799 farmers in six central provinces of Turkey were checked for the presence of Rathayibacter species by plating 100 µl of the diluted and undiluted seed wash suspension onto modified 523 agar. Of the 25 isolated strains presumptively identified as Rathayibac...

Accurate Determination of Soluble Axl by Enzyme-Linked Immunosorbent Assay.

PubMed

Dengler, Mirko; Huber, Heidemarie; Müller, Christian J; Zellmer, Angela; Rauch, Peter; Mikulits, Wolfgang

2016-11-01

Levels of soluble Axl (sAxl) are routinely assessed in human sera by sandwich enzyme-linked immunosorbent assay (ELISA). Although sAxl values are suggested to diagnose different types of disorders, no uniform ELISA method is available, allowing the reliable interassay comparison between results. Furthermore, little is known about the stability of sAxl under storage conditions, which is a relevant parameter for biomedical trials. The evaluation of sAxl stability under various stress conditions and the determination of proper conditions to use the sAxl ELISA for routine clinical applications are of great interest. In this study, serum samples were subjected to freeze-thaw cycles and incubation at different temperatures to analyze the stability of sAxl by ELISA. Dilution and spike-in experiments were carried out to examine the impact of serum and diluent components on the ELISA performance. Various diluents and media were employed to resolve masking effects of the serum. The assay components were further optimized for long-term usability by treatment with stabilizers and validation under temperature stress. Indeed, sAxl showed long-term stability in serum during freeze-thaw cycles and incubation under temperature stress conditions. The dilution experiments revealed that unknown components in the serum caused masking effects that can be reduced by proper dilutions. The assay performance was further increased by using a standardized buffer system to dilute serum samples. Stabilization of coated plates and of streptavidin-horseradish peroxidase allowed long-term storage for up to 6 months. In sum, our data demonstrate proper ELISA conditions, allowing the accurate analysis of sAxl levels in human serum.

A rapid 'one-plate' in vitro test for pyrogens.

PubMed

Poole, Stephen; Mistry, Yogesh; Ball, Christine; Gaines Das, Rose E; Opie, Lisa Pieti; Tucker, Garry; Patel, Mehul

2003-03-01

A rapid, 'one-plate' monocyte-activation test is described for detecting endotoxin and non-endotoxin pyrogens in parenteral medicinal products. The one-plate test offers useful gains over conventional 'two-plate' (cell culture plate+ELISA plate) tests in terms of its limit of detection, robustness, speed and cost. The 'one-plate' test is likely to be applicable to a wide range of products because it allows less time for product interference in the test. The 'one-plate' test utilises pyrogen-free anti-cytokine (interleukin (IL)-6 or tumour necrosis factor alpha (TNFalpha)) antibodies (Ab), coated and stabilised onto (pyrogen-free) 96-well plates. Monocytes/monocytic cells, endotoxin (lipopolysaccharides, LPS) standard or sample and (pyrogen-free) second (labelled) Ab are cultured together (usually for 2-4 h) on the Ab-coated plate and then the plate is washed and the ELISA completed. There is no transfer from one plate to another and no (further) incubations of (released) cytokine with, first, coating Ab and, then, developing Ab since these steps have already taken place during the initial cell culture. The rapid, 'one-plate' test is readily automated. The preferred readout is IL-6, which gives a limit of detection of 0.015 endotoxin units (EU)/ml with peripheral blood mononuclear cell (PBMNC), 0.03 EU/ml with diluted whole blood and 0.05 EU/ml with a monocytic cell line (MONO MAC 6).

The Resazurin-Agar Method - a Quick Test to Determine Water Quality

NASA Astrophysics Data System (ADS)

Huckfeldt, J.; Westphal, B.; Claußen, L.

2015-12-01

Rezasurin has been used as a smart tracer in stream ecosystems to indicate metabolic activity, specifically aerobic respiration by heterotrophic bacteria. Resazurin is a blue compound which is irreversibly reduced to the pink resorufin in the presence of aerobic bacteria. The degree and speed of colour change from blue to pink is a measure of the degree of oxygen consumption and thus an indirect indication of the concentration of aerobic bacteria in a given medium. A high concentration of bacteria in water indicates a bad water quality. In our work a method was developed using resazurin agar plates to find a quick and easy way for testing water quality and comparing concentrations of bacteria in freshwater and seawater samples. The theory was to concentrate bacteria from a defined volume of water sample onto polycarbonate filters (0.2 μm), which are then placed onto the resazurin agar plate. The presence of aerobic bacteria on the filter will reduce the resazurin in the agar and the compound changes its colour. First tests conducted with different dilutions of a pure culture of yoghurt bacteria showed promising results and confirmed the feasibility of the method. In a further assay, we used water samples from different water layers and different temperatures and were also able to observe differences in the concentration of bacteria, depending on these different environmental conditions.The assay was also successfully used with seawater samples, collected from 2 different stations at 3 different depths in the Baltic Sea (salinity=15). The discolouration of the plates showed good correlation with the oxygen concentrations in the water. The resazurin-agar plate method is economical and fast. Several samples could be investigated at the same time without sacrificing the reliability of the results. Thus it is a good pre-screening test for a quantitative evaluation of bacteria in a water sample.

Microbial contamination in intraoral phosphor storage plates: the dilemma.

PubMed

de Souza, Tricia Murielly Pereira Andrade; de Castro, Ricardo Dias; de Vasconcelos, Laís César; Pontual, Andréa Dos Anjos; de Moraes Ramos Perez, Flávia Maria; Pontual, Maria Luiza Dos Anjos

2017-01-01

The aims of this study were to evaluate microbial contamination in phosphor storage plates in dental radiology services and discuss the possible origin of this contamination. The sample comprised 50 phosphor plates: 14 plates from service A, 30 from service B, and 6 in the control group, consisting of plates never used. Damp sterile swabs were rubbed on the phosphor plates, and then transferred to tests tubes containing sterile saline solution. Serial dilutions were made, and then inoculated in triplicate on Mueller Hinton agar plates and incubated at 37 °C/48 h, before counting the colony-forming units (CFU). The samples were also seeded in brain-heart infusion medium to confirm contamination by turbidity of the culture medium. All solutions, turbid and clean, were seeded in selective and non-selective media. At service A and B, 50 and 73.3 % of the phosphor plates were contaminated, respectively. This contamination was mainly due to bacteria of the genus Staphylococcus. CFU counts ranged from 26.4 to 80.0 CFU/plate. Most of the phosphor plates evaluated shown to be contaminated, mainly by Staphylococcus ssp. Quantitatively, this contamination occurred at low levels, possibly arising from handling of the plates. The use of a second plastic barrier may have diminished contamination by microorganisms from the oral cavity. There is a risk of cross-contamination by phosphor storage plates used in dental radiology services.

Effects of aspect ratio and concentration on rheology of epoxy suspensions containing model plate-like nanoparticles

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

White, K. L.; Takahara, A.; Institute for Materials Chemistry and Engineering, Kyushu University, Fukuoka 819-0395

2015-12-15

Hexagonal 2-dimensional α-zirconium phosphate crystals were prepared with lateral diameters ranging from 110 nm to 1.5 μm to investigate the effect of particle size on suspension rheology. The nanoplatelets were exfoliated to individual sheets with monodisperse thickness and dispersed in a Newtonian epoxy fluid. The steady shear response of dilute and semi-dilute suspensions was measured and compared to expressions obtained from theory for infinitely dilute suspensions. For suspensions containing the smaller nanoplatelets, aspect ratio ∼160, the low shear rate viscosity and transition to shear thinning behavior were well described by theory for loadings up to 0.5 vol. %. The agreementmore » was improved by assuming a moderate polydispersity in lateral diameter, ∼30%–50%, which is consistent with experimental observation. For the higher aspect ratio nanoplatelets, good agreement between theory and experiment was observed only at high shear rates. At lower shear rate, theory consistently over-predicted viscosity, which was attributed to a progressive shift to non-isotropic initial conditions with increasing particle size. The results suggest that at a fixed Peclet number, there is an increasing tendency for the nanoplatelets to form transient, local stacks as particle size increases. The largest particles, aspect ratio ∼2200, showed unusual shear thinning and thickening behaviors that were attributed to particle flexibility. The findings demonstrate the surprising utility of theory for infinitely dilute suspensions to interpret, and in some cases quantitatively describe, the non-Newtonian viscosity of real suspensions containing high aspect ratio plate-like particles. A simple framework is proposed to interpret deviations from ideal behavior based on the local and collective behavior of the suspended nanoplatelets.« less

Optimised method to estimate octanol water distribution coefficient (logD) in a high throughput format.

PubMed

Low, Ying Wei Ivan; Blasco, Francesca; Vachaspati, Prakash

2016-09-20

Lipophilicity is one of the molecular properties assessed in early drug discovery. Direct measurement of the octanol-water distribution coefficient (logD) requires an analytical method with a large dynamic range or multistep dilutions, as the analyte's concentrations span across several orders of magnitude. In addition, water/buffer and octanol phases which have very different polarity could lead to matrix effects and affect the LC-MS response, leading to erroneous logD values. Most compound libraries use DMSO stocks as it greatly reduces the sample requirement but the presence of DMSO has been shown to underestimate the lipophilicity of the analyte. The present work describes the development of an optimised shake flask logD method using deepwell 96 well plate that addresses the issues related to matrix effects, DMSO concentration and incubation conditions and is also amenable to high throughput. Our results indicate that the equilibrium can be achieved within 30min by flipping the plate on its side while even 0.5% of DMSO is not tolerated in the assay. This study uses the matched matrix concept to minimise the errors in analysing the two phases namely buffer and octanol in LC-MS. Copyright © 2016 Elsevier B.V. All rights reserved.

Development of large area nanostructured silicon-hydrogen alloy material with improved stability for solar cell application by argon dilution method

NASA Astrophysics Data System (ADS)

Dey, Arka; Das, Mrinmay; Datta, Joydeep; Jana, Rajkumar; Dhar, Joydeep; Sil, Sayantan; Biswas, Debasish; Banerjee, Chandan; Ray, Partha Pratim

2016-07-01

Here we have presented the results of large area (30 × 30 cm2) silicon-hydrogen alloy material and solar cell by argon dilution method. As an alternative to hydrogen dilution, argon dilution method has been applied to develop single junction solar cell with appreciable stability. Optimization of deposition conditions revealed that 95% argon dilution gives a nanostructured material with improved transport property and less light induced degradation. The minority carrier diffusion length (L d ) and mobility-lifetime (μτ) product of the material with 95% argon dilution degrades least after light soaking. Also the density of states (DOS) below conduction level reveals that this material is less defective. Solar cell with this argon diluted material has been fabricated with all the layers deposited by argon dilution method. Finally we have compared the argon diluted solar cell results with the optimized hydrogen diluted solar cell. Light soaking study proves that it is possible to develop stable solar cell on large area by argon dilution method and that the degradation of argon diluted solar cell is less than that of hydrogen diluted one. [Figure not available: see fulltext.

Effect of Carbon Dioxide on Testing of Susceptibilities of Respiratory Tract Pathogens to Macrolide and Azalide Antimicrobial Agents

PubMed Central

Johnson, M. M.; Hill, S. L.; Piddock, Laura J. V.

1999-01-01

The in vitro activities of erythromycin, azithromycin, and clarithromycin against 178 clinical isolates from the lower respiratory tract of patients with chronic obstructive pulmonary disease were determined by an agar dilution method. The plates were incubated in air alone or in 5% carbon dioxide. The MICs measured in air alone were lower for most isolates than those measured in 5% carbon dioxide, illustrating the “pH effect” of incubation in carbon dioxide. Testing of isolates in 5% carbon dioxide on pH-adjusted medium (pH 8.4) resulted in MICs of one or two doubling dilutions lower than those obtained on agar with a neutral pH. A bioassay of the three agents incubated in air and in 5% carbon dioxide resulted in a significant loss of activity of all three agents in the carbon dioxide-enriched atmosphere. However, this loss-of-activity effect was significantly reduced when the bioassay medium was adjusted to pH 8.4 prior to incubation in 5% carbon dioxide. PMID:10428903

[The effect of biyuanshu oral liquid on the formation of Pseudomonas aeruginosa biofilms in vitro].

PubMed

Liu, Xiang; Chen, Haihong; Wang, Shengqing

2012-07-01

To observe the effect of biyuanshu oral liquid on the formation of pseudomonas aeruginosa biofilms in vitro. Pseudomonas aeruginosa biofilm was established by plate culture and detected by Scanning electron microscopy and AgNO3 staining. After treated with different dosages of biyuanshu oral liquid and erythromycin, the pseudomonas aeruginosa biofilms were observed by AgNO3 staining and the number of viable bacteria were measured by serial dilution. The pseudomonas aeruginosa biofilms could be detected by SEM at the seventh culture day and it was consistent with the detection of AgNO3 staining. The biyuanshu oral liquid and erythromycin have the effect on inhibiting the formation of pseudomonas aeruginosa biofilms. But with the already formed pseudomonas aeruginosa biofilms the inhibition was not significant. The serial dilution method showed that the viable counts of bacteria of biyuanshu oral liquid and erythromycin treated groups were significantly lower than those untreated groups (P < 0.05). The biyuanshu oral liquid and erythromycin can inhibit the formation of pseudomonas aeruginosa biofilms in vitro.

Automation of the anthrone assay for carbohydrate concentration determinations.

PubMed

Turula, Vincent E; Gore, Thomas; Singh, Suddham; Arumugham, Rasappa G

2010-03-01

Reported is the adaptation of a manual polysaccharide assay applicable for glycoconjugate vaccines such as Prevenar to an automated liquid handling system (LHS) for improved performance. The anthrone assay is used for carbohydrate concentration determinations and was scaled to the microtiter plate format with appropriate mixing, dispensing, and measuring operations. Adaptation and development of the LHS platform was performed with both dextran polysaccharides of various sizes and pneumococcal serotype 6A polysaccharide (PnPs 6A). A standard plate configuration was programmed such that the LHS diluted both calibration standards and a test sample multiple times with six replicate preparations per dilution. This extent of replication minimized the effect of any single deviation or delivery error that might have occurred. Analysis of the dextran polymers ranging in size from 214 kDa to 3.755 MDa showed that regardless of polymer chain length the hydrolysis was complete, as evident by uniform concentration measurements. No plate positional absorbance bias was observed; of 12 plates analyzed to examine positional bias the largest deviation observed was 0.02% percent relative standard deviation (%RSD). The high purity dextran also afforded the opportunity to assess LHS accuracy; nine replicate analyses of dextran yielded a mean accuracy of 101% recovery. As for precision, a total of 22 unique analyses were performed on a single lot of PnPs 6A, and the resulting variability was 2.5% RSD. This work demonstrated the capability of a LHS to perform the anthrone assay consistently and a reduced assay cycle time for greater laboratory capacity.

Expeditious neutralization assay for human metapneumovirus based on a recombinant virus expressing Renilla luciferase.

PubMed

Zhou, Min; Kitagawa, Yoshinori; Yamaguchi, Mayu; Uchiyama, Chika; Itoh, Masae; Gotoh, Bin

2013-01-01

Human metapneumovirus (HMPV) is a common cause of respiratory diseases in persons of all ages. Because of its slow replication and weak cytopathic effect in cultured cells, conventional neutralization assays for HMPV require around one week for completion. The purpose of this study is to establish a rapid neutralization assay based on a recombinant virus expressing Renilla luciferase (Rluc). A recombinant HMPV expressing both Rluc and green fluorescent protein (GFP) was created by reverse genetics method. Two-fold serial dilutions of human 23 sera were made in a 96-well plate and incubated with 50 pfu/well of the recombinant virus at 4°C for 1 h. The mixtures were then transferred to LLC-MK2 cells in a 96-well plate, incubated for 2 h, and replaced with trypsin-free fresh media. After incubation at 32°C for 24 h, the cells were lysed and measured for Rluc activity. The neutralization titer was defined as the reciprocal of the highest serum dilution that resulted in 50% reduction of Rluc activity. The novel assay could be completed within 24 h and eliminated the requirement of trypsin supporting multistep replication in cultured cells, as well as laborious processes including the plaque assay with immunostaining. Neutralization titers correlated well with those determined by a GFP-based assay previously developed. The neutralization assay based on Rluc activity is the fastest and the most straightforward of all previous assays, and may be available for high throughput screening of neutralizing antibodies. Copyright © 2012 Elsevier B.V. All rights reserved.

Evaluation of culture-based techniques and 454 pyrosequencing for the analysis of fungal diversity in potting media and organic fertilizers.

PubMed

Al-Sadi, A M; Al-Mazroui, S S; Phillips, A J L

2015-08-01

Potting media and organic fertilizers (OFs) are commonly used in agricultural systems. However, there is a lack of studies on the efficiency of culture-based techniques in assessing the level of fungal diversity in these products. A study was conducted to investigate the efficiency of seven culture-based techniques and pyrosequencing for characterizing fungal diversity in potting media and OFs. Fungal diversity was evaluated using serial dilution, direct plating and baiting with carrot slices, potato slices, radish seeds, cucumber seeds and cucumber cotyledons. Identity of all the isolates was confirmed on the basis of the internal transcribed spacer region of the ribosomal RNA (ITS rRNA) sequence data. The direct plating technique was found to be superior over other culture-based techniques in the number of fungal species detected. It was also found to be simple and the least time consuming technique. Comparing the efficiency of direct plating with 454 pyrosequencing revealed that pyrosequencing detected 12 and 15 times more fungal species from potting media and OFs respectively. Analysis revealed that there were differences between potting media and OFs in the dominant phyla, classes, orders, families, genera and species detected. Zygomycota (52%) and Chytridiomycota (60%) were the predominant phyla in potting media and OFs respectively. The superiority of pyrosequencing over cultural methods could be related to the ability to detect obligate fungi, slow growing fungi and fungi that exist at low population densities. The evaluated methods in this study, especially direct plating and pyrosequencing, may be used as tools to help detect and reduce movement of unwanted fungi between countries and regions. © 2015 The Society for Applied Microbiology.

Paper-based device for separation and cultivation of single microalga.

PubMed

Chen, Chih-Chung; Liu, Yi-Ju; Yao, Da-Jeng

2015-12-01

Single-cell separation is among the most useful techniques in biochemical research, diagnosis and various industrial applications. Microalgae species have great economic importance as industrial raw materials. Microalgae species collected from environment are typically a mixed and heterogeneous population of species that must be isolated and purified for examination and further application. Conventional methods, such as serial dilution and a streaking-plate method, are intensive of labor and inefficient. We developed a paper-based device for separation and cultivation of single microalga. The fabrication was simply conducted with a common laser printer and required only a few minutes without lithographic instruments and clean-room. The driving force of the paper device was simple capillarity without a complicated pump connection that is part of most devices for microfluidics. The open-structure design of the paper device makes it operable with a common laboratory micropipette for sample transfer and manipulation with a naked eye or adaptable to a robotic system with functionality of high-throughput retrieval and analysis. The efficiency of isolating a single cell from mixed microalgae species is seven times as great as with a conventional method involving serial dilution. The paper device can serve also as an incubator for microalgae growth on simply rinsing the paper with a growth medium. Many applications such as highly expressed cell selection and various single-cell analysis would be applicable. Copyright © 2015 Elsevier B.V. All rights reserved.

Study of methods for the improvement of bacterial transport media

NASA Technical Reports Server (NTRS)

Gardner, R. L.; Beakley, J. W.

1973-01-01

A series of 500 transport media recipes was tested for ability to hold pure cultures of Streptococcus equisimilus, Corynebacterium equi, Neisseria perflava, and Haemophilus parainfluenzae for 21 days. Stuart Medium Base with 0.4% agar was used as the control medium for this and the other experiments in the investigation. At the end of the holding period inoculated transport media were quantitatively assayed, and the control media were assayed immediately after inoculation. Three vials of each medium were inoculated with an organism, and each vial's medium was diluted and spread on duplicate plates. Assay media for this experiment included Brain Heart Infusion,(BHIA) Tryptic Soy Agar, and BHIA with 1% Isovitalex enrichment.

[Antimicrobial susceptibility testing of anaerobic bacteria].

PubMed

García-Sánchez, José E; García-Sánchez, Enrique; García-García, María Inmaculada

2014-02-01

The anaerobic bacteria resistance to antibiotics is increasing, and even has appeared against the most active of those, like metronidazol and carbapenems. This fact forces to make and periodical sensibility tests -at least in the most aggressive and virulent species, in cases that they are isolated from life locations and in the absence of therapeutic response- to check the local sensibility and to establish suitable empiric therapies, all based on multicentric studies carried out in order to this or well to check the activity of new antibiotics. For the laboratory routine, the easiest sensibility method is the E-test/MIC evaluator. Another alternative is microdilution, that's only normalized for Bacteroides. There are preliminary facts that allow the use of disc diffusion method in some species of Bacteroides and Clostridium. For the temporal and multicentric studies, the procedure is dilution in agar plate, the reference method. Copyright © 2014 Elsevier España, S.L. All rights reserved.

Synthetic Nanovaccines Against Respiratory Pathogens (SYNARP)

DTIC Science & Technology

2013-09-01

destroying enzyme ( RDE ) (Denka-Seiken, Tokyo, Japan) prior to being tested. Briefly, three parts RDE was added to one part sera and incubated...overnight at 37°C. RDE was inactivated by incubation at 56°C for ~60 min. RDE -treated sera were two-fold serially diluted in v-bottom microtiter plates. An

Interfering with DNA Damage Signals: Radiosensitizing Prostate Cancer Using Small Peptides

DTIC Science & Technology

2009-05-01

25-l sample of the supernatant was analyzed for free phosphate in the malachite green assay by dilution with 100 l of a developing solution... malachite green). After incubation for 15 min, the release of phosphate was quantified by measuring the absorbance at 650 nm in a microtiter plate reader

Detection of Phytophthora ramorum chlamydospores in soil by baiting and dilution plating

USDA-ARS?s Scientific Manuscript database

Chlamydospores of P. ramorum produced by mixing 20 percent V8 juice broth cultures with sand and incubating over a 1 month period were used to infest field soil at concentrations ranging from 0.2 to 42 chlamydospores/cc soil. Chlamydospore recovery was determined by baiting with rhododendron leaf d...

Evaluation and optimization of compound solubilization and delivery methods in a two-tiered ion channel lead optimization triage.

PubMed

Hendricson, Adam W; Gallagher, Liz; Matchett, Michele; Ferrante, Meredith; Spence, Steve; Paiva, Tony; Shou, Wilson; Tertyshnikova, Svetlana; Krambis, Mike; Post-Munson, Deborah; Zhang, Litao; Knox, Ron

2012-04-01

Low-volume dispensing of neat dimethyl sulfoxide (DMSO) into plate-based assays conserves compound, assay reagents, and intermediate dilution plate cost and, as we demonstrate here, significantly improves structure-activity relationship resolution. Acoustic dispensing of DMSO solutions into standard volume 384W plates yielded inconsistent results in studies with 2 cell lines because of apparent effects on the integrity of the cell monolayer (increased intracellular Ca⁺⁺ levels as indicated by elevated basal dye fluorescence after acoustic transfer). PocketTip-mediated transfer was successful at increasing apparent potency on a more consistent basis. Notably, the correlation coefficient among fluorescence imaging plate reader (FLIPR):electrophysiology (EP) across a representative ~125 compound collection was increased ~5× via conversion to a PocketTip direct dispensation, indicating a triage assay more predictive of activity in the decisional patch-clamp assay. Very importantly, the EP-benchmarked false-negative rate as measured by compounds with FLIPR EC₅₀ more than the highest concentration tested fell from >11% to 5% assay-wide, and the relative FLIPR:EP rank-order fidelity increased from 55% to 78%. Elimination of the aqueous intermediate step provided additional benefits, including reduced assay cost, decreased cycle time, and reduced wet compound consumption rate. Direct DMSO dispensing has broad applicability to cell-based functional assays of multiple varieties, especially in cases where limit solubility in assay buffer is a recognized impediment to maximizing interassay connectivity.

Oligonucleotide Array for Identification and Detection of Pythium Species†

PubMed Central

Tambong, J. T.; de Cock, A. W. A. M.; Tinker, N. A.; Lévesque, C. A.

2006-01-01

A DNA array containing 172 oligonucleotides complementary to specific diagnostic regions of internal transcribed spacers (ITS) of more than 100 species was developed for identification and detection of Pythium species. All of the species studied, with the exception of Pythium ostracodes, exhibited a positive hybridization reaction with at least one corresponding species-specific oligonucleotide. Hybridization patterns were distinct for each species. The array hybridization patterns included cluster-specific oligonucleotides that facilitated the recognition of species, including new ones, belonging to groups such as those producing filamentous or globose sporangia. BLAST analyses against 500 publicly available Pythium sequences in GenBank confirmed that species-specific oligonucleotides were unique to all of the available strains of each species, of which there were numerous economically important ones. GenBank entries of newly described species that are not putative synonyms showed no homology to sequences of the spotted species-specific oligonucleotides, but most new species did match some of the cluster-specific oligonucleotides. Further verification of the specificity of the DNA array was done with 50 additional Pythium isolates obtained by soil dilution plating. The hybridization patterns obtained were consistent with the identification of these isolates based on morphology and ITS sequence analyses. In another blind test, total DNA of the same soil samples was amplified and hybridized on the array, and the results were compared to those of 130 Pythium isolates obtained by soil dilution plating and root baiting. The 13 species detected by the DNA array corresponded to the isolates obtained by a combination of soil dilution plating and baiting, except for one new species that was not represented on the array. We conclude that the reported DNA array is a reliable tool for identification and detection of the majority of Pythium species in environmental samples. Simultaneous detection and identification of multiple species of soilborne pathogens such as Pythium species could be a major step forward for epidemiological and ecological studies. PMID:16597974

[Use of bacteriphages against Salmonella Enteritidis: a prevention tool].

PubMed

García, Cristina; Marín, Clara; Catalá-Gregori, Pablo; Soriano, Jose Miguel

2015-06-01

Salmonellosis is a highly prevalent disease still searching for preventive tools to avoid contamination level priority public health. The in vitro effect of bacteriophages against Salmonella enteritidis was evaluated as a prevention tool. Two tests with three concentrations of bacteriophages were conducted against two strains of Salmonella Enteritidis inoculated in fresh faeces of laying hens. Each test had a positive control. Thus, four groups in each test were evaluated. Each experimental group included two replicates, and three plates were incubated per replicate. The concentrations tested were three: commercial solution (5 × 10(7) pfu/mL), and two dilutions (1/10 and 1/30). One of the strains tested was CECT 4300, a certified strain of Colección Española de Cultivo Tipo and the other a field isolated strain in a sacrificed hen farm. Both strains were inoculated at 1.3 × 10(5) cfu/g of faeces in each of the four groups. Isolation and identification of bacteria by ISO6579 was done at various times after inoculation: 1 minute, 24 hours and 7 days. In the first test, with certified strain, Salmonella was isolated in all groups at time 1 minute. After 24 hours, Salmonella was isolated in all groups except in one of the replicas treated with 1/10 dilution of bacteriophages, one of the other replica plate treated with 1/10 dilution, and two plates of the two replicas treated with the commercial solution. After 7 days, the bacteria were not isolated from any of the experimental groups. In the second test, with the field strain, Salmonella was isolated in all groups at time 1 minute. After 24 hours, Salmonella was isolated in all groups except in one of the replicas treated with 1/10 dilution of bacteriophages and the two replicas treated with the commercial solution. Salmonella was not isolated in any of the experimental groups at 7 days. The use of bacteriophages reduced Salmonella enteritidis isolates in faeces at 24 hours after the application, so it could be considered as a prevention tool. At 7 days after inoculation of bacteria, no one was isolated in any of the experimental groups. Copyright AULA MEDICA EDICIONES 2014. Published by AULA MEDICA. All rights reserved.

Evaluation of ATP measurements to detect microbial ingress by wastewater and surface water in drinking water.

PubMed

Vang, Óluva K; Corfitzen, Charlotte B; Smith, Christian; Albrechtsen, Hans-Jørgen

2014-11-01

Fast and reliable methods are required for monitoring of microbial drinking water quality in order to protect public health. Adenosine triphosphate (ATP) was investigated as a potential real-time parameter for detecting microbial ingress in drinking water contaminated with wastewater or surface water. To investigate the ability of the ATP assay in detecting different contamination types, the contaminant was diluted with non-chlorinated drinking water. Wastewater, diluted at 10(4) in drinking water, was detected with the ATP assay, as well as 10(2) to 10(3) times diluted surface water. To improve the performance of the ATP assay in detecting microbial ingress in drinking water, different approaches were investigated, i.e. quantifying microbial ATP or applying reagents of different sensitivities to reduce measurement variations; however, none of these approaches contributed significantly in this respect. Compared to traditional microbiological methods, the ATP assay could detect wastewater and surface water in drinking water to a higher degree than total direct counts (TDCs), while both heterotrophic plate counts (HPC 22 °C and HPC 37 °C) and Colilert-18 (Escherichia coli and coliforms) were more sensitive than the ATP measurements, though with much longer response times. Continuous sampling combined with ATP measurements displays definite monitoring potential for microbial drinking water quality, since microbial ingress in drinking water can be detected in real-time with ATP measurements. The ability of the ATP assay to detect microbial ingress is influenced by both the ATP load from the contaminant itself and the ATP concentration in the specific drinking water. Consequently, a low ATP concentration of the specific drinking water facilitates a better detection of a potential contamination of the water supply with the ATP assay. Copyright © 2014 Elsevier Ltd. All rights reserved.

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.

Oxyrase, a method which avoids CO2 in the incubation atmosphere for anaerobic susceptibility testing of antibiotics affected by CO2.

PubMed

Spangler, S K; Appelbaum, P C

1993-02-01

The Oxyrase agar dilution method, with exclusion of CO2 from the environment, was compared with the reference agar dilution method recommended by the National Committee for Clinical Laboratory Standards (anaerobic chamber with 10% CO2) to test the susceptibility of 51 gram-negative and 43 gram-positive anaerobes to azithromycin and erythromycin. With the Oxyrase method, anaerobiosis was achieved by incorporation of the O2-binding enzyme Oxyrase in addition to susceptibility test medium, antibiotic, and enzyme substrates into the upper level of a biplate. Plates were covered with a Brewer lid and incubated in ambient air. With azithromycin, Oxyrase yielded an MIC for 50% of strains tested (MIC50) and MIC90 of 2.0 and 8.0 micrograms/ml, compared to 8.0 and > 32.0 micrograms/ml in standard anaerobic conditions. At a breakpoint of 8.0 micrograms/ml, 90.4% of strains were susceptible to azithromycin with Oxyrase, compared to 53.2% in the chamber. The corresponding erythromycin MIC50 and MIC90 were 1.0 and 8.0 micrograms/ml with Oxyrase, compared to 4.0 and > 32.0 micrograms/ml by the reference method, with 89.3% of strains susceptible at a breakpoint of 4 micrograms/ml with Oxyrase, compared to 60.6% in CO2. Exclusion of CO2 from the anaerobic atmosphere when testing for susceptibility to azalides and macrolides yielded lower MICs, which may lead to a reconsideration of the role played by these compounds in treatment of infections caused by these strains.

A fluorescent microplate assay for diarrheic shellfish toxins.

PubMed

Vieytes, M R; Fontal, O I; Leira, F; Baptista de Sousa, J M; Botana, L M

1997-06-01

A fluorescent enzyme inhibition assay for okadaic acid using 4-methylumbelliferyl phosphate and fluorescein diphosphate as substrates for the enzyme phosphatase 2A was developed. In the inhibition assay, performed in a microtiter plate, the PP2A was inhibited by adding okadaic acid and the resulting fluorescence enhancement derived from enzymatic hydrolysis of the substrate was quantified in a fluorescence plate reader. The measurable range of okadaic acid was 3.2 to 3200 pg/ml with an IC50 = 0.1 nM. The detection limit of okadaic acid was 2.56 pg/well in buffer solutions and 12.8 ng/g hepatopancreas in shellfish extracts. The coefficient of variation (CV, n = 22) for each point ranged from 18.80 to 37.90% (mean 28.35%). The proposed method is very convenient, rapid, and sensitive by using the enzyme inhibition assay system and fluorescent reaction as a detection system. This work demonstrates that the fluorescent assay can be used to quantify the amount of okadaic acid in shellfish samples and also is valid for very dilute samples, such as phytoplankton samples.

Fun Microbiology: Using a Plant Pathogenic Fungus To Demonstrate Koch's Postulates.

ERIC Educational Resources Information Center

Mitchell, James K.; Orsted, Kathy M.; Warnes, Carl E.

1997-01-01

Describes an experiment using a plant pathogenic fungus in which students learn to follow aseptic techniques, grow and produce spores of a fungus, use a hemacytometer for enumerating spores, prepare serial dilutions, grow and inoculate plants, isolate a pure culture using agar streak plates, and demonstrate the four steps of Koch's postulates.…

Optimization and validation of a high throughput method for detecting neutralizing antibodies against human papillomavirus (HPV) based on pseudovirons.

PubMed

Nie, Jianhui; Huang, Weijin; Wu, Xueling; Wang, Youchun

2014-09-01

The pseudoviron-based neutralization assay is accepted as the gold standard to evaluate the functional humoral immune response against HPV. The goal of this study was to develop and optimize a human papillomavirus (HPV) neutralization assay using HPV pseudovirons with Gaussia luciferase (Gluc) as the reporter gene. For this purpose, high-titers Gluc pseudovirons were generated by cotransfecting 293TT cells with HPV structural genes and Gluc expressing plasmids. Six types of neutralizing monoclonal antibodies, vaccines immunized serum samples and WHO international antibody standard were used to validate the new developed assay. The ideal circumstances of the assay were identified for cell counts (30,000/well for 96-well plate), pseudoviron inoculating size (100 times RLU above background) and incubation time (72 hr). The sensitivity of the Gluc assay was comparable to secreted alkaline phosphatase (SEAP) assay and higher than the green florescent protein (GFP) assay. The non-specific background for different types of sample was significantly different (rabbit sera > human sera > mouse sera, P < 0.01). The non-specific neutralization effects were not attributed to IgG antibody. The cutoff value for this assay was determined as 50% inhibition at a dilution of 1:40. Without requirements of sample dilution and different incubation times at different temperature before processing, the detection time was shortened from more than 90 min to less than 5 min for a 96-well plate compared with the SEAP-based assay. With the advantages of short detection time and easy-to-use procedure, the newly developed assay is more suitable for large sero-epidemiological studies or clinical trials and more amenable to automation. © 2014 Wiley Periodicals, Inc.

Effects of various heavy metal nanoparticles on Enterococcus hirae and Escherichia coli growth and proton-coupled membrane transport.

PubMed

Vardanyan, Zaruhi; Gevorkyan, Vladimir; Ananyan, Michail; Vardapetyan, Hrachik; Trchounian, Armen

2015-10-16

Due to bacterial resistance to antibiotics there is a need for new antimicrobial agents. In this respect nanoparticles can be used as they have expressed antibacterial activity simultaneously being more reactive compared to their bulk material. The action of zinc (II), titanium (IV), copper (II) and (I) oxides thin films with nanostructured surface and silver nanoscale particles on Enterococcus hirae and Escherichia coli growth and membrane activity was studied by using microbiological, potentiometric and spectrophotometric methods. It was revealed that sapphire base plates with deposited ZnO, TiO2, CuO and Cu2O nanoparticles had no effects neither on E. hirae nor E. coli growth both on agar plates and in liquid medium. Concentrated Ag nanoparticles colloid solution markedly affected bacterial growth which was expressed by changing growth properties. E. hirae was able to grow only at <1:200 dilutions of Ag nanoparticles while E. coli grew even at 1:10 dilution. At the same time Ag nanoparticles directly affected membranes, as the FOF1-ATPase activity and H(+)-coupled transport was changed either (E. coli were less susceptible to nanoparticles compared to E. hirae). Ag nanoparticles increased H(+) and K(+) transport even in the presence of N,N'-dicyclohexylcarbodiimide (DCCD), inhibitor of FOF1. The stoichiometry of DCCD-inhibited ion fluxes was disturbed. These results point out to distinguishing antibacterial effects of Ag nanoparticles on different bacteria; the difference between effects can be explained by peculiarities in bacterial membrane structure and properties. H(+)-K(+)-exchange disturbance by Ag nanoparticles might be involved in antibacterial effects on E. hirae. The role of FOF1 in antibacterial action of Ag nanoparticles was shown using atpD mutant lacked β subunit in F1.

A miniaturized fibrinolytic assay for plasminogen activators

NASA Technical Reports Server (NTRS)

Lewis, M. L.; Nachtwey, D. S.; Damron, K. L.

1991-01-01

This report describes a micro-clot lysis assay (MCLA) for evaluating fibrinolytic activity of plasminogen activators (PA). Fibrin clots were formed in wells of microtiter plates. Lysis of the clots by PA, indicated by change in turbidity (optical density, OD), was monitored with a microplate reader at five minutes intervals. Log-log plots of PA dilution versus endpoint, the time at which the OD value was halfway between the maximum and minimum value for each well, were linear over a broad range of PA concentrations (2-200 International units/ml). The MCLA is a modification and miniaturization of well established fibrinolytic methods. The significant practical advantages of the MCLA are that it is a simple, relatively sensitive, non-radioactive, quantitative, kinetic, fibrinolytic micro-technique which can be automated.

Relative Numbers of Certain Microbial Groups Present in Compost Used for Mushroom (Agaricus bisporus) Propagation

PubMed Central

Fordyce, C.

1970-01-01

The relative numbers of microorganisms associated with compost during mushroom production were studied by the dilution plate method. Thermophilic actinomycetes and fungi were isolated with a very high frequency early in the growing season. Although numbers of thermophilic bacteria diminished slowly during the season, the thermophilic fungi and actinomycetes diminished rapidly with the latter disappearing after 6 weeks. Mesophilic fungi other than Agaricus or Trichoderma remained relatively stable throughout the growing period. Agaricus could be isolated between the first and third break. Trichoderma became dominant after the fourth break. The mesophilic bacterial counts diminished during the most productive portion of the mushroom cropping season and then increased to much higher numbers toward the end of the season. PMID:5529631

High-Throughput Quantification of Bacterial-Cell Interactions Using Virtual Colony Counts

PubMed Central

Hoffmann, Stefanie; Walter, Steffi; Blume, Anne-Kathrin; Fuchs, Stephan; Schmidt, Christiane; Scholz, Annemarie; Gerlach, Roman G.

2018-01-01

The quantification of bacteria in cell culture infection models is of paramount importance for the characterization of host-pathogen interactions and pathogenicity factors involved. The standard to enumerate bacteria in these assays is plating of a dilution series on solid agar and counting of the resulting colony forming units (CFU). In contrast, the virtual colony count (VCC) method is a high-throughput compatible alternative with minimized manual input. Based on the recording of quantitative growth kinetics, VCC relates the time to reach a given absorbance threshold to the initial cell count using a series of calibration curves. Here, we adapted the VCC method using the model organism Salmonella enterica sv. Typhimurium (S. Typhimurium) in combination with established cell culture-based infection models. For HeLa infections, a direct side-by-side comparison showed a good correlation of VCC with CFU counting after plating. For MDCK cells and RAW macrophages we found that VCC reproduced the expected phenotypes of different S. Typhimurium mutants. Furthermore, we demonstrated the use of VCC to test the inhibition of Salmonella invasion by the probiotic E. coli strain Nissle 1917. Taken together, VCC provides a flexible, label-free, automation-compatible methodology to quantify bacteria in in vitro infection assays. PMID:29497603

Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells.

PubMed

Fedr, Radek; Pernicová, Zuzana; Slabáková, Eva; Straková, Nicol; Bouchal, Jan; Grepl, Michal; Kozubík, Alois; Souček, Karel

2013-05-01

The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples. Copyright © 2013 International Society for Advancement of Cytometry.

Evaluation of Petrifilm Lactic Acid Bacteria Plates for Counting Lactic Acid Bacteria in Food.

PubMed

Kanagawa, Satomi; Ohshima, Chihiro; Takahashi, Hajime; Burenqiqige; Kikuchi, Misato; Sato, Fumina; Nakamura, Ayaka; Mohamed, Shimaa M; Kuda, Takashi; Kimura, Bon

2018-06-01

Although lactic acid bacteria (LAB) are used widely as starter cultures in the production of fermented foods, they are also responsible for food decay and deterioration. The undesirable growth of LAB in food causes spoilage, discoloration, and slime formation. Because of these adverse effects, food companies test for the presence of LAB in production areas and processed foods and consistently monitor the behavior of these bacteria. The 3M Petrifilm LAB Count Plates have recently been launched as a time-saving and simple-to-use plate designed for detecting and quantifying LAB. This study compares the abilities of Petrifilm LAB Count Plates and the de Man Rogosa Sharpe (MRS) agar medium to determine the LAB count in a variety of foods and swab samples collected from a food production area. Bacterial strains isolated from Petrifilm LAB Count Plates were identified by 16S rDNA sequence analysis to confirm the specificity of these plates for LAB. The results showed no significant difference in bacterial counts measured by using Petrifilm LAB Count Plates and MRS medium. Furthermore, all colonies growing on Petrifilm LAB Count Plates were confirmed to be LAB, while yeast colonies also formed in MRS medium. Petrifilm LAB Count Plates eliminated the plate preparation and plate inoculation steps, and the cultures could be started as soon as a diluted food sample was available. Food companies are required to establish quality controls and perform tests to check the quality of food products; the use of Petrifilm LAB Count Plates can simplify this testing process for food companies.

Inactivation of Mold Spores from Moist Carpet Using Steam Vapor: Contact Time and Temperature.

PubMed

Ong, Kee-Hean; Emo, Brett; Lewis, Roger D; Kennedy, Jason; Thummalakunta, Laxmi N A; Elliott, Michael

2015-01-01

Steam vapor has been shown to reduce viable mold spores in carpet, but the minimal effective temperature and contact time has not been established. This study evaluated the effectiveness of steam vapor in reducing the number of viable mold spores in carpet as a function of temperature and contact time. Seventy carpet samples were inoculated with a liquid suspension of Cladosporium sphaerospermum and incubated over a water-saturated foam carpet pad for 24 hr. Steam was applied to the samples as the temperature was measured from the carpet backing. Contact time was closely monitored over seven time intervals: 0, 2, 4, 8, 12, 16, and 20 sec. Following steam vapor treatment, mold spores were extracted from the carpet samples and the extract was plated on DG-18 plates at 1:1, 1:10, 1:100 dilutions followed by one week of incubation. Raw colony forming units were determined using an automated colony counter and adjusted based on dilution factor, extraction volume, and plated volume. Analysis of variance and linear regression were used to test for statistically significant relationships. Steam contact time exhibited a linear relationship to observed temperature of carpet backing (F = 90.176, R(2) = 0.609). Observed temperature of carpet backing had a positive relationship to percent reduction of mold (F = 76.605, R(2) = 0.569). Twelve seconds of steam vapor contact time was needed to achieve over 90% mold reduction on moist carpet.

Effect of Iranian Ziziphus honey on growth of some foodborne pathogens

PubMed Central

Ekhtelat, Maryam; Ravaji, Karim; Parvari, Masood

2016-01-01

Background: Honey has previously been shown to have wound healing and antimicrobial properties, but this is dependent on the type of honey, geographical location, and flower from which the final product is derived. We tested the antimicrobial activity of a natural honey originating from the Ziziphus spina-christi tree, against selected strains of bacteria. Ziziphus honey among more than a 100 verities of honey is known to have the greatest value of energy and minerals in it. Materials and Methods: The aim of this study was to determine the antibacterial activity of Ziziphus honey in 10%, 20%, 30%, and 40% dilutions (v/v) against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus. Viable count enumeration of the sample was investigated after 0, 24, 72, and 120 h postinoculation with any of the bacteria using pour-plate method. Results: The findings indicate that Ziziphus honey was effective against these pathogenic bacteria. In a comparative trial, antibacterial activity of Ziziphus honey was higher after 120 h incubation for each four bacteria in most dilutions. The microbial count showed 3-7.5 log reduction comparing with control after 120 h. Conclusions: Therefore, it is recommended using Ziziphus honey as a natural preservative and antibacterial agent. Also, it could potentially be used as therapeutic agents against bacterial infection particularly to the tested microorganisms. PMID:27003970

A novel agar formulation for isolation and direct enumeration of Vibrio vulnificus from oyster tissue.

PubMed

Griffitt, Kimberly J; Grimes, D Jay

2013-08-01

A new selective and differential medium, Vibrio vulnificus X-Gal (VVX), was developed for direct enumeration of V. vulnificus (Vv) from oyster samples. This agar utilizes cellobiose and lactose as carbon sources, and the antibiotics colistin and polymyxin B as selective agents. Hydrolysis of 5-bromo-4-chloro-3-indolyl- beta-d-galactopyranoside (x-gal), used in the agar as a lactose analog, produces an insoluble blue dye that makes lactose positive colonies easily distinguishable from any non-lactose fermenting bacteria. Various bacterial species were spot plated onto thiosulfate-citrate-bile salts-sucrose agar (TCBS), and CHROMagar Vibrio, two vibrio-specific selective agars, non-selective agar, and VVX to compare selectivity of VVX to other widely used media. A V. vulnificus pure culture was serially diluted on VVX and non-selective agar to determine the VVX percent recovery. Water and oyster samples were spread plated on VVX agar and allowed to incubate for 16-18 h at 33 °C. Blue and white colonies from VVX agar were picked and screened by end point PCR for the Vv hemolysin vvhA. VVX agar showed a significant improvement over TCBS and CHROMagar at preventing non-target growth. There was an 87.5% recovery compared to non-selective plating and a 98% positivity rate of blue colonies picked from oyster tissue plating. The findings suggest that this new agar is a fast, distinctive, and accurate method for enumeration of V. vulnificus from the environment. Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical and biological characterization of novel essential oils from Eremophila bignoniiflora (F. Muell) (Myoporaceae): a traditional Aboriginal Australian bush medicine.

PubMed

Sadgrove, Nicholas John; Hitchcock, Maria; Watson, Kenneth; Jones, Graham Lloyd

2013-10-01

Essential oils were extracted by hydrodistillation from the traditional Australian medicinal plant Eremophila bignoniiflora, characterized chemically and then screened for bioactivity. Characterization and quantification were completed using gas chromatography-mass spectrometry (GC-MS) and GC-flame ionization detection, respectively. Antimicrobial capacity was assessed using disc diffusion and micro-titre plate broth dilution and further characterized using thin layer chromatography followed by bioautography to assign activity to separated individual active components. Antifungal capacity was investigated using micro-titre plate broth dilution against pathogenic Trichophyton species. Free radical scavenging ability was assessed using the diphenylpicrylhydradyl reaction in methanol. The predominant components of the essential oil were fenchyl-acetate and bornyl-acetate. However, bioautography indicated antimicrobial ability to be largely linked to the less abundant, more polar constituents. Oils displayed only modest antifungal ability against pathogenic Trichophyton species associated with dermatophytosis, but moderate to high antimicrobial activity, particularly against the yeast Candida albicans and the bacteria Staphylococcus epidermidis. Essential oils exhibited relatively low free radical scavenging ability. Speculation over the role of essential oils in the traditional medicinal applications of E. bignoniiflora follows, exploring correlations between traditional use and investigated bioactivities. Copyright © 2012 John Wiley & Sons, Ltd.

Total body water and lean body mass estimated by ethanol dilution

NASA Technical Reports Server (NTRS)

Loeppky, J. A.; Myhre, L. G.; Venters, M. D.; Luft, U. C.

1977-01-01

A method for estimating total body water (TBW) using breath analyses of blood ethanol content is described. Regression analysis of ethanol concentration curves permits determination of a theoretical concentration that would have existed if complete equilibration had taken place immediately upon ingestion of the ethanol; the water fraction of normal blood may then be used to calculate TBW. The ethanol dilution method is applied to 35 subjects, and comparison with a tritium dilution method of determining TBW indicates that the correlation between the two procedures is highly significant. Lean body mass and fat fraction were determined by hydrostatic weighing, and these data also prove compatible with results obtained from the ethanol dilution method. In contrast to the radioactive tritium dilution method, the ethanol dilution method can be repeated daily with its applicability ranging from diseased individuals to individuals subjected to thermal stress, strenuous exercise, water immersion, or the weightless conditions of space flights.

Dilutions Made Easy.

ERIC Educational Resources Information Center

Kamin, Lawrence

1996-01-01

Presents problems appropriate for high school and college students that highlight dilution methods. Promotes an understanding of dilution methods in order to prevent the unnecessary waste of chemicals and glassware in biology laboratories. (JRH)

Development of a Cold Sterilant for Field Medical Use.

DTIC Science & Technology

1996-11-01

Dental Devices Using STERIS 20C at Use Dilution and Manual Soak Technique .............. 1086 Akron Rubber Development Laboratory Report 32918...simulated reuse protocol with lot P-2 of 25 months of age , a number of positives (i.e. sterilization failures) were noted, in particular for the...Ethylene propylene ELASTOMERS Brass Ethylene propylene diene Ethylene propylene diene ( EPDM ) ( EPDM ) Brass 360 Polyamide RTV 133 Nickel-plated Cooper

Chemiluminescent optical fiber immunosensor for the detection of anti-West Nile virus IgG.

PubMed

Herrmann, Sebastien; Leshem, Boaz; Landes, Shimi; Rager-Zisman, Bracha; Marks, Robert S

2005-03-31

An ELISA-based optical fiber methodology developed for the detection of anti-West Nile virus IgG antibodies in serum was compared to standard colorimetric and chemiluminescent ELISA based on microtiter plates. Colorimetric ELISA was the least sensitive, especially at high titer dilutions. The fiber-optic immunosensor based on the same ELISA immunological rationale was the most sensitive technique.

[Analysis on the antimicrobial resistance of lactic acid bacteria isolated from the yogurt sold in China].

PubMed

Fan, Qin; Liu, Shuliang; Li, Juan; Huang, Tingting

2012-05-01

To analyze the antimicrobial susceptibility of lactic acid bacteria (LAB) from yogurt, and to provide references for evaluating the safety of LAB and screening safe strains. The sensitivity of 43 LAB strains, including 14 strains of Streptococcus thermophilus, 12 strains of Lactobacillus acidophilus, 9 strains of Lactobacillus bulgaricus and 8 strains of Bifidobacterium, to 22 antibiotics were tested by agar plate dilution method. All 43 LAB strains were resistant to trimethoprim, nalidixic acid, ciprofloxacin, lomefloxacin, danofloxacin and polymyxin E. Their resistances to kanamycin, tetracycline, clindamycin, doxycycline and cephalothin were varied. The sensitivity to other antibiotics were sensitive or moderate. All isolates were multidrug-resistant. The antimicrobial resistance of tested LAB strains was comparatively serious, and continuously monitoring their antimicrobial resistance and evaluating their safety should be strengthened.

Successful enrichment of the ubiquitous freshwater acI Actinobacteria.

PubMed

Garcia, Sarahi L; McMahon, Katherine D; Grossart, Hans-Peter; Warnecke, Falk

2014-02-01

Actinobacteria of the acI lineage are often the numerically dominant bacterial phylum in surface freshwaters, where they can account for > 50% of total bacteria. Despite their abundance, there are no described isolates. In an effort to obtain enrichment of these ubiquitous freshwater Actinobacteria, diluted freshwater samples from Lake Grosse Fuchskuhle, Germany, were incubated in 96-well culture plates. With this method, a successful enrichment containing high abundances of a member of the lineage acI was established. Phylogenetic classification showed that the acI Actinobacteria of the enrichment belonged to the acI-B2 tribe, which seems to prefer acidic lakes. This enrichment grows to low cell densities and thus the oligotrophic nature of acI-B2 was confirmed. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

Laboratory evaluation of 3M Petrifilms and University of Minnesota Bi-plates as potential on-farm tests for clinical mastitis.

PubMed

McCarron, J L; Keefe, G P; McKenna, S L B; Dohoo, I R; Poole, D E

2009-05-01

The objective was to determine test characteristics and compare 2 potential on-farm culture systems for clinical mastitis, the Minnesota Easy Culture System II Bi-plate and Petrifilm. The tests were evaluated using clinically positive mastitic milk samples (n = 282) to determine their ability to differentiate appropriate treatment groups; all cases that had gram-positive growth were considered treatment candidates (n = 161), whereas cases that grew gram-negative organisms only or yielded no bacterial growth were classified as no treatment (n = 121). For Petrifilm, both undiluted and 1:10 diluted milk samples were used. To create treatment categories, 2 types of Petrifilms were used, Aerobic Count (AC) and Coliform Count (CC). Both Bi-plates and Petrifilms were read after 24 h of incubation. Analysis was conducted at various colony count thresholds for the Petrifilm test system. The combination of Petrifilms that had the highest sensitivity classified a case as gram-negative if there were > or =20 colonies present on the CC. If there were <20 colonies present on the CC and >5 colonies present on the AC, a case would be classified as gram-positive. The Bi-plate had a sensitivity of 97.9% and a specificity of 68.6%. The Petrifilm test system had a sensitivity of 93.8% and a specificity of 70.1%. There was no significant difference in the sensitivities between the tests. All Bi-plates and Petrifilms were read by a laboratory technician and a group of masked readers with limited microbiology training. Kappa values for the masked readers were 0.75 for Bi-plates and 0.84 and 0.86 for AC and CC Petrifilms, respectively. The Bi-plate and Petrifilm were able to successfully categorize clinical cases of mastitis into 2 treatments based on their ability to detect the presence of a gram-positive organism. Neither method had the ability to determine if a sample was contaminated. The results of this study indicate that both tests were able to appropriately categorize cases, which could potentially result in a reduction in the quantity of antibiotics used to treat clinical cases of mastitis.

Solvent shift method for anti-precipitant screening of poorly soluble drugs using biorelevant medium and dimethyl sulfoxide.

PubMed

Yamashita, Taro; Ozaki, Shunsuke; Kushida, Ikuo

2011-10-31

96-well plate based anti-precipitant screening using bio-relevant medium FaSSIF (fasted-state simulated small intestinal fluid) is a useful technique for discovering anti-precipitants that maintain supersaturation of poorly soluble drugs. In a previous report, two disadvantages of the solvent evaporation method (solvent casting method) were mentioned: precipitation during the evaporation process and the use of volatile solvents to dissolve compounds. In this report, we propose a solvent shift method using DMSO (dimethyl sulfoxide). Initially, the drug substance was dissolved in DMSO at a high concentration and diluted with FaSSIF that contained anti-precipitants. To evaluate the validity of the method, itraconazole (ITZ) was used as the poorly soluble model drug. The solvent shift method resolved the disadvantages of the evaporation method, and AQOAT (HPMC-AS) was found as the most appropriate anti-precipitant for ITZ in a facile and expeditious manner when compared with the solvent evaporation method. In the large scale JP paddle method, AQOAT-based solid dispersion maintained a higher concentration than Tc-5Ew (HPMC)-based formulation; this result corresponded well with the small scale of the solvent shift method. Copyright © 2011 Elsevier B.V. All rights reserved.

Disentangling niche competition from grazing mortality in phytoplankton dilution experiments

PubMed Central

Weitz, Joshua S.

2017-01-01

The dilution method is the principal tool used to infer in situ microzooplankton grazing rates. However, grazing is the only mortality process considered in the theoretical model underlying the interpretation of dilution method experiments. Here we evaluate the robustness of mortality estimates inferred from dilution experiments when there is concurrent niche competition amongst phytoplankton. Using a combination of mathematical analysis and numerical simulations, we find that grazing rates may be overestimated—the degree of overestimation is related to the importance of niche competition relative to microzooplankton grazing. In response, we propose a conceptual method to disentangle the effects of niche competition and grazing by diluting out microzooplankton, but not phytoplankton. Our theoretical results suggest this revised “Z-dilution” method can robustly infer grazing mortality, regardless of the dominant phytoplankton mortality driver in our system. Further, we show it is possible to independently estimate both grazing mortality and niche competition if the classical and Z-dilution methods can be used in tandem. We discuss the significance of these results for quantifying phytoplankton mortality rates; and the feasibility of implementing the Z-dilution method in practice, whether in model systems or in complex communities with overlap in the size distributions of phytoplankton and microzooplankton. PMID:28505212

Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry

NASA Technical Reports Server (NTRS)

Pyle, B. H.; Broadaway, S. C.; McFeters, G. A.

1999-01-01

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0. 95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.

Production of Alkaline Cellulase by Fungi Isolated from an Undisturbed Rain Forest of Peru

PubMed Central

Vega, Karin; Villena, Gretty K.; Sarmiento, Victor H.; Ludeña, Yvette; Vera, Nadia; Gutiérrez-Correa, Marcel

2012-01-01

Alkaline cellulase producing fungi were isolated from soils of an undisturbed rain forest of Peru. The soil dilution plate method was used for the enumeration and isolation of fast growing cellulolytic fungi on an enriched selective medium. Eleven out of 50 different morphological colonies were finally selected by using the plate clearing assay with CMC as substrate at different pH values. All 11 strains produced cellulases in liquid culture with activities at alkaline pH values without an apparent decrease of them indicating that they are true alkaline cellulase producers. Aspergillus sp. LM-HP32, Penicillium sp. LM-HP33, and Penicillium sp. LM-HP37 were the best producers of FP cellulase (>3 U mL−1) with higher specific productivities (>30 U g−1 h−1). Three strains have been found suitable for developing processes for alkaline cellulase production. Soils from Amazonian rain forests are good sources of industrial fungi with particular characteristics. The results of the present study are of commercial and biological interest. Alkaline cellulases may be used in the polishing and washing of denim processing of the textile industry. PMID:23213539

[Bacteriological study on juvenile periodontitis].

PubMed

Han, N

1991-02-01

The predominant cultivable microflora of 23 pockets in 15 juvenile periodontitis (JP) patients was studied for the first time in China using the current anaerobic methodology. Samples were taken with sterile paper points and dispersed on a vortex mixer. Then the diluted samples were plated on the non-selective blood agar plates and selective MGB medium which favors the growth of Actinobacillus actimycetemcomitans (Aa) and incubated in anaerobic chamber for 5 days. From each sample 15 or more isolated colonies were picked in sequence without selection and subcultured. The isolates were identified mainly by Schrechenberger's 4 hour rapid methods for biochemical and fermentative tests and the chromatographic analysis of acid end products using ion-chromatography. The results were as follows: 1. The microflora of healthy sulci of 7 healthy young subjects was significantly different from that in the pocket of JP patients. The predominant species in healthy sulci were Streptococcus spp and Capnocytophaga gingivalis. 2. The species increased significantly in JP patients in prevalence and proportions was Eubacterium. Other species in high proportions were Bacteroides oris, B. melaninogenicus, B. gingivalis, Capnocytophaga sputigena, and Actinomyces meyeri, etc. 3. Actinobacillus actinomycetemcomitans was not detected in any of the samples.

Comparative determination of two probiotics by QCM and OWLS-based immunosensors.

PubMed

Szalontai, Helga; Adányi, Nóra; Kiss, Attila

2014-09-25

The regular consumption of foods containing probiotic bacteria has beneficial physiological effects on the health and the digestion system. There is a need for novel analytical approaches for the determination of these bacteria that are faster than the classical plate counting method. For this purpose, two label-free biosensors were investigated and presented in this paper: Quartz Crystal Microbalance (QCM) and Optical Waveguide Lightmode Spectroscopy (OWLS) based direct immunosensors were developed for real-time direct detection of probiotic bacteria in fermented dairy products. Bifidobacterium bifidum O1356 and Lactobacillus acidophilus O1132 were detected by polyclonal anti-B. bifidum IgG and anti-L. acidophilus IgG immobilized on the sensors' surface. Sulfo-LC-SPDP cross linking agent was used to bind antibodies to the gold surface of the QCM's AT-cut quartz wafer. Concerning OWLS, antibodies were covalently bound to the amino groups of the silanized surface of the waveguide by glutaraldehyde. The dynamic measuring range was found between 1.0E+3 and 5.0E+5CFUmL(-1) in 100 fold diluted fermented milk products by QCM and with OWLS. Considering the current legislation of the probiotic content in probiotic products, the two developed immunosensors can be applied for rapid quantification of L. acidophilus and B. bifidum in fermented milk. These examinations offer effective alternatives to the microbiological plate counting method. Copyright © 2014 Elsevier B.V. All rights reserved.

A new gas dilution method for measuring body volume.

PubMed Central

Nagao, N; Tamaki, K; Kuchiki, T; Nagao, M

1995-01-01

This study was designed to examine the validity of a new gas dilution method (GD) for measuring human body volume and to compare its accuracy with the results obtained by the underwater weighing method (UW). We measured the volume of plastic bottles and 16 subjects (including two females), aged 18-42 years with each method. For the bottles, the volume measured by hydrostatic weighing was correlated highly (r = 1.000) with that measured by the new gas dilution method. For the subjects, the body volume determined by the two methods was significantly correlated (r = 0.998). However, the subject's volume measured by the gas dilution method was significantly larger than that by underwater weighing method. There was significant correlation (r = 0.806) between GD volume-UW volume and the body mass index (BMI), so that UW volume could be predicted from GD volume and BMI. It can be concluded that the new gas dilution method offers promising possibilities for future research in the population who cannot submerge underwater. PMID:7551760

Comparison of quartz tuning forks and AlN-based extensional microresonators for viscosity measurements in oil/fuel mixtures

NASA Astrophysics Data System (ADS)

Toledo, J.; Manzaneque, T.; Hernando-García, J.; Vazquez, J.; Ababneh, A.; Seidel, H.; Lapuerta, M.; Sánchez-Rojas, J. L.

2013-05-01

In-situ monitoring of the physical properties of liquids is of great interest in the automotive industry. For example, lubricants are subject to dilution with diesel fuel as a consequence of late-injection processes, which are necessary for regenerating diesel particulate filters. This dilution can be determined by tracking the viscosity and the density of the lubricant. Here we report the test of two in-plane movement based resonators to explore their capability to monitor oil dilution with diesel and biodiesel. One of the resonators is the commercially available millimeter-sized quartz tuning fork, working at 32.7 kHz. The second resonator is a state-of-the-art micron-sized AlN-based rectangular plate, actuated in the first extensional mode in the MHz range. Electrical impedance measurements were carried out to characterize the performance of the structures in various liquid media in a wide range of viscosities. These measurements were completed with the development of low-cost electronic circuits to track the resonance frequency and the quality factor automatically, these two parameters allow to obtain the viscosity of various fluids under investigation, as in the case of dilution of lubricant SAE 15W40 and biodiesel.

Gradient, contact-free volume transfers minimize compound loss in dose-response experiments.

PubMed

Harris, David; Olechno, Joe; Datwani, Sammy; Ellson, Richard

2010-01-01

More accurate dose-response curves can be constructed by eliminating aqueous serial dilution of compounds. Traditional serial dilutions that use aqueous diluents can result in errors in dose-response values of up to 4 orders of magnitude for a significant percentage of a compound library. When DMSO is used as the diluent, the errors are reduced but not eliminated. The authors use acoustic drop ejection (ADE) to transfer different volumes of model library compounds, directly creating a concentration gradient series in the receiver assay plate. Sample losses and contamination associated with compound handling are therefore avoided or minimized, particularly in the case of less water-soluble compounds. ADE is particularly well suited for assay miniaturization, but gradient volume dispensing is not limited to miniaturized applications.

Split-plot designs for robotic serial dilution assays.

PubMed

Buzas, Jeffrey S; Wager, Carrie G; Lansky, David M

2011-12-01

This article explores effective implementation of split-plot designs in serial dilution bioassay using robots. We show that the shortest path for a robot to fill plate wells for a split-plot design is equivalent to the shortest common supersequence problem in combinatorics. We develop an algorithm for finding the shortest common supersequence, provide an R implementation, and explore the distribution of the number of steps required to implement split-plot designs for bioassay through simulation. We also show how to construct collections of split plots that can be filled in a minimal number of steps, thereby demonstrating that split-plot designs can be implemented with nearly the same effort as strip-plot designs. Finally, we provide guidelines for modeling data that result from these designs. © 2011, The International Biometric Society.

Discovery of potent KIFC1 inhibitors using a method of integrated high-throughput synthesis and screening.

PubMed

Yang, Bin; Lamb, Michelle L; Zhang, Tao; Hennessy, Edward J; Grewal, Gurmit; Sha, Li; Zambrowski, Mark; Block, Michael H; Dowling, James E; Su, Nancy; Wu, Jiaquan; Deegan, Tracy; Mikule, Keith; Wang, Wenxian; Kaspera, Rüdiger; Chuaqui, Claudio; Chen, Huawei

2014-12-11

KIFC1 (HSET), a member of the kinesin-14 family of motor proteins, plays an essential role in centrosomal bundling in cancer cells, but its function is not required for normal diploid cell division. To explore the potential of KIFC1 as a therapeutic target for human cancers, a series of potent KIFC1 inhibitors featuring a phenylalanine scaffold was developed from hits identified through high-throughput screening (HTS). Optimization of the initial hits combined both design-synthesis-test cycles and an integrated high-throughput synthesis and biochemical screening method. An important aspect of this integrated method was the utilization of DMSO stock solutions of compounds registered in the corporate compound collection as synthetic reactants. Using this method, over 1500 compounds selected for structural diversity were quickly assembled in assay-ready 384-well plates and were directly tested after the necessary dilutions. Our efforts led to the discovery of a potent KIFC1 inhibitor, AZ82, which demonstrated the desired centrosome declustering mode of action in cell studies.

Effect of Iranian Ziziphus honey on growth of some foodborne pathogens.

PubMed

Ekhtelat, Maryam; Ravaji, Karim; Parvari, Masood

2016-01-01

Honey has previously been shown to have wound healing and antimicrobial properties, but this is dependent on the type of honey, geographical location, and flower from which the final product is derived. We tested the antimicrobial activity of a natural honey originating from the Ziziphus spina-christi tree, against selected strains of bacteria. Ziziphus honey among more than a 100 verities of honey is known to have the greatest value of energy and minerals in it. The aim of this study was to determine the antibacterial activity of Ziziphus honey in 10%, 20%, 30%, and 40% dilutions (v/v) against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus. Viable count enumeration of the sample was investigated after 0, 24, 72, and 120 h postinoculation with any of the bacteria using pour-plate method. The findings indicate that Ziziphus honey was effective against these pathogenic bacteria. In a comparative trial, antibacterial activity of Ziziphus honey was higher after 120 h incubation for each four bacteria in most dilutions. The microbial count showed 3-7.5 log reduction comparing with control after 120 h. Therefore, it is recommended using Ziziphus honey as a natural preservative and antibacterial agent. Also, it could potentially be used as therapeutic agents against bacterial infection particularly to the tested microorganisms.

Heavy-metal resistance in Gram-negative bacteria isolated from Kongsfjord, Arctic.

PubMed

Neethu, C S; Mujeeb Rahiman, K M; Saramma, A V; Mohamed Hatha, A A

2015-06-01

Isolation and characterization of heterotrophic Gram-negative bacteria was carried out from the sediment and water samples collected from Kongsfjord, Arctic. In this study, the potential of Arctic bacteria to tolerate heavy metals that are of ecological significance to the Arctic (selenium (Se), mercury (Hg), cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn)) was investigated. Quantitative assay of 130 isolates by means of plate diffusion and tube dilution methods was carried out by incorporation of different concentrations of metals. Growth in Se and Pb at a concentration of 3000 μg/L was significantly lower (P≤0.0001) than at 2000 μg/L. The minimum inhibitory concentration for Cd and Hg was 50 μg/L (P≤0.0001, F=264.23 and P≤0.0001, F=291.08, respectively) even though in the tube dilution test, Hg-containing tubes showed much less growth, revealing its superior toxicity to Cd. Thus, the level of toxicity of heavy metals was found to be in the order of Hg>Cd>Cu>Zn>Pb>Se. Multiple-metal-resistant isolates were investigated for their resistance against antibiotics, and a positive correlation was observed between antibiotic and metal resistance for all the isolates tested. The resistant organisms thus observed might influence the organic and inorganic cycles in the Arctic and affect the ecosystem.

Draft Genome Sequences of Two Janthinobacterium lividum Strains, Isolated from Pristine Groundwater Collected from the Oak Ridge Field Research Center

PubMed Central

Wu, Xiaoqin; Deutschbauer, Adam M.; Kazakov, Alexey E.; Wetmore, Kelly M.; Cwick, Bryson A.; Walker, Robert M.; Novichkov, Pavel S.; Arkin, Adam P.

2017-01-01

ABSTRACT We present here the draft genome sequences of two Janthinobacterium lividum strains, GW456P and GW458P, isolated from groundwater samples collected from a background site at the Oak Ridge Field Research Center. Production of a purple pigment by these two strains was observed when grown on diluted (1/10) LB agar plates. PMID:28663297

Method and apparatus for maintaining multi-component sample gas constituents in vapor phase during sample extraction and cooling

DOEpatents

Farthing, William Earl [Pinson, AL; Felix, Larry Gordon [Pelham, AL; Snyder, Todd Robert [Birmingham, AL

2008-02-12

An apparatus and method for diluting and cooling that is extracted from high temperature and/or high pressure industrial processes. Through a feedback process, a specialized, CFD-modeled dilution cooler is employed along with real-time estimations of the point at which condensation will occur within the dilution cooler to define a level of dilution and diluted gas temperature that results in a gas that can be conveyed to standard gas analyzers that contains no condensed hydrocarbon compounds or condensed moisture.

Method and apparatus maintaining multi-component sample gas constituents in vapor phase during sample extraction and cooling

DOEpatents

Farthing, William Earl; Felix, Larry Gordon; Snyder, Todd Robert

2009-12-15

An apparatus and method for diluting and cooling that is extracted from high temperature and/or high pressure industrial processes. Through a feedback process, a specialized, CFD-modeled dilution cooler is employed along with real-time estimations of the point at which condensation will occur within the dilution cooler to define a level of dilution and diluted gas temperature that results in a gas that can be conveyed to standard gas analyzers that contains no condensed hydrocarbon compounds or condensed moisture.

In Vitro Comparison of Terbinafine and Itraconazole against Penicillium marneffei

PubMed Central

McGinnis, Michael R.; Nordoff, Nicole G.; Ryder, Neil S.; Nunn, Gary B.

2000-01-01

We evaluated terbinafine and itraconazole against 30 isolates of Penicillium marneffei using a modification of the National Committee for Clinical Laboratory Standards broth macrodilution MIC testing protocol for yeasts. The minimal fungicidal concentration (MFC) was determined by plating 100 μl from each MIC drug dilution having no growth onto Sabouraud glucose agar incubated at 30°C. The MFC was the dilution at which growth was absent at 72 h of incubation. The MICs, in micrograms per milliliter, were as follows: terbinafine, 0.03 to 1.0 (geometric mean titer, 0.09); itraconazole, 0.03 to 0.5 (geometric mean titer, 0.04). The MFCs, in micrograms per milliliter, were as follows: terbinafine, 0.03 to 8 (geometric mean titer, 2.60); itraconazole, 0.03 to 8 (geometric mean titer, 2.45). Primary fungicidal activity (MFC within 2 dilutions of MIC) was observed with terbinafine in eight isolates and with itraconazole in four isolates. The data indicate that terbinafine is active against P. marneffei in vitro and may have a previously unrealized role in the management of infections caused by this fungus. PMID:10770792

Effect of brewery wastewater obtained from different phases of treatment plant on seed germination of chickpea (Cicer arietinum), maize (Zea mays), and pigeon pea (Cajanus cajan).

PubMed

Salian, Rupa; Wani, Suhas; Reddy, Ramamohan; Patil, Mukund

2018-03-01

Brewing industry releases large quantities of wastewater after product generation. Brewery wastewater contains organic compounds which are biodegradable in nature. These biodegradable wastes can be recycled and reused and hence considered as suitable products for agriculture. But before using wastewater for agriculture, it is better to evaluate the phytotoxic effects of wastewater on crops. Hence, the main objective of this study is to evaluate the effects of brewery effluent on seed germination and growth parameters of selected crop species like chickpea (Cicer arietinum), maize (Zea mays), and pigeon pea (Cajanus cajan). Study comprised seven types of water treatments-tap water as control, diluted UASBR effluent (50% effluent + 50% distilled water): UASBR50, undiluted UASBR effluent: UASBR100, diluted TC effluent (50% effluent + 50% distilled water): ETP50,TC effluent without dilution: ETP100, 10% diluted reverse osmosis (RO10) reject (10% RO reject + 90% distilled water), and 25% diluted reverse osmosis(RO25) reject (25% RO reject + 75% distilled water) with three replications in completely randomized design. Germination test was performed in petri plates for 5 days. Parameters like germination percentage, germination rate index, seedling length, phytotoxicity index, seed vigor index, and biomass were calculated. All parameters decreased with increase in respective effluent concentration. Among all treatments, RO25 showed highest inhibitory effect on all three crops. Even though undiluted effluent of UASBR and ETP effluent showed positive effect on germination, seedling growth of three crops was promoted to the maximum by UASBR50 and ETP50. Hence, from the study, it was concluded that dilution of brewery effluent can be recommended before using it for irrigational purpose.

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.

Effect of IFN-gamma on the killing of S. aureus in human whole blood. Assessment of bacterial viability by CFU determination and by a new method using alamarBlue.

PubMed

DeForge, L E; Billeci, K L; Kramer, S M

2000-11-01

Given the increasing incidence of methicillin resistant Staphylococcus aureus (MRSA) and the recent emergence of MRSA with a reduced susceptibility to vancomycin, alternative approaches to the treatment of infection are of increasing relevance. The purpose of these studies was to evaluate the effect of IFN-gamma on the ability of white blood cells to kill S. aureus and to develop a simpler, higher throughput bacterial killing assay. Using a methicillin sensitive clinical isolate of S. aureus, a clinical isolate of MRSA, and a commercially available strain of MRSA, studies were conducted using a killing assay in which the bacteria were added directly into whole blood. The viability of the bacteria in samples harvested at various time points was then evaluated both by the classic CFU assay and by a new assay using alamarBlue. In the latter method, serially diluted samples and a standard curve containing known concentrations of bacteria were placed on 96-well plates, and alamarBlue was added. Fluorescence readings were taken, and the viability of the bacteria in the samples was calculated using the standard curve. The results of these studies demonstrated that the CFU and alamarBlue methods yielded equivalent detection of bacteria diluted in buffer. For samples incubated in whole blood, however, the alamarBlue method tended to yield lower viabilities than the CFU method due to the emergence of a slower growing subpopulation of S. aureus upon incubation in the blood matrix. A significant increase in bacterial killing was observed upon pretreatment of whole blood for 24 h with 5 or 25 ng/ml IFN-gamma. This increase in killing was detected equivalently by the CFU and alamarBlue methods. In summary, these studies describe a method that allows for the higher throughput analysis of the effects of immunomodulators on bacterial killing.

[Isolation and fermentation condition of milk-clotting enzyme producing strain from glutinous rice wine].

PubMed

Cheng, Qiaoling; Bai, Xiaojia; Wang, Yanping

2008-06-01

Glutinous rice wine is a traditional food in south of China and it can coagulate milk. It has been proved that its function of coagulating milk is because of the presence of milk-clotting enzyme produced by microorganisms in glutinous rice wine. The aim of this work is to isolate milk-clotting enzyme producing strain from glutinous rice wine and study the fermentation condition. We screened out four bacteria and fungus by gradient dilution. It was proved that mold played the most important role in the production of milk-clotting enzyme. This is further confirmed by casein plate method. The optimization of fermentation conditions revealed that two times concentrated potato medium supplemented with 5% glucose without additional nitrogen was better for production of the enzyme. The enzyme activity was increased 144% under the conditions established.

Serological activity of white-tail deer against several species of Brucella.

PubMed

Salinas-Meléndez, J A; Martínez-Muñoz, A; Avalos-Ramírez, R; Cerutti-Pereyra, N; Riojas-Valdés, V M

1998-01-01

In Mexico, brucellosis is a widely distributed disease of domesticated ruminants, but its frequency in wild ruminants has not been documented. Since northeast Mexico is the main distribution area of white-tailed deer and has been reported as an area positive for brucellosis in domesticated species, the present study was conducted in order to determine serological activity against several species of the genus Brucella in white-tailed deer. A total of 208 sera of white-tailed deer were collected during the springs of 1994 and 1995 in the north part of the states of Nuevo León and Coahuila. Each serum was analyzed for the detection of antibodies against two smooth (B. abortus and B. melitensis) and one rough (B. ovis) species of the genus Brucella. The serological tests used for the determination of the presence of antibodies against Brucella were card and plate agglutination for B. abortus, plate agglutination and rivanol precipitation for B. melitensis, and agar gel immunodiffusion for B. ovis. Each assay had positive and negative controls. None of the analyzed samples was found to be positive, and only two sera showed partial plate agglutination against B. melitensis at a dilution of 1:25; however, at higher dilutions and to the rivanol precipitation test the same samples were negative. Therefore, the percentage of positive sera was estimated at 0% (0/208). This result makes evident the absence of positive white-tailed deer against Brucella in the sampled area, despite that this disease is considered present in domesticated species. Therefore, white-tailed deer does not have, at the present time, an important role for the dispersion of the disease. The same result has been reported in other countries.

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.

A screening method for cardiovascular active compounds in marine algae.

PubMed

Agatonovic-Kustrin, S; Kustrin, E; Angove, M J; Morton, D W

2018-05-18

The interaction of bioactive compounds from ethanolic extracts of selected marine algae samples, separated on chromatographic plates, with nitric/nitrous acid was investigated. The nature of bioactive compounds in the marine algae extracts was characterised using UV absorption spectra before and after reaction with diluted nitric acid, and from the characteristic colour reaction after derivatization with anisaldehyde. It was found that diterpenes from Dictyota dichotoma, an edible brown algae, and sterols from green algae Caulerpa brachypus, bind nitric oxide and may act as a nitric oxide carrier. Although the carotenoid fucoxanthin, found in all brown marine algae also binds nitric oxide, the bonds between nitrogen and the fucoxanthin molecule are much stronger. Further studies are required to evaluate the effects of diterpenes from Dictyota dichotoma and sterols from green algae Caulerpa brachypus to see if they have beneficial cardiovascular effects. The method reported here should prove useful in screening large numbers of algae species for compounds with cardiovascular activity. Copyright © 2018 Elsevier B.V. All rights reserved.

Correlation potential of a test ion near a strongly charged plate.

PubMed

Lu, Bing-Sui; Xing, Xiangjun

2014-03-01

We analytically calculate the correlation potential of a test ion near a strongly charged plate inside a dilute m:-n electrolyte. We do this by calculating the electrostatic Green's function in the presence of a nonlinear background potential, the latter having been obtained using the nonlinear Poisson-Boltzmann theory. We consider the general case where the dielectric constants of the plate and the electrolyte are distinct. The following generic results emerge from our analyses: (1) If the distance to the plate Δz is much larger than a Gouy-Chapman length, the plate surface will behave effectively as an infinitely charged surface, and the dielectric constant of the plate effectively plays no role. (2) If Δz is larger than a Gouy-Chapman length but shorter than a Debye length, the correlation potential can be interpreted in terms of an image charge that is three times larger than the source charge. This behavior is independent of the valences of the ions. (3) The Green's function vanishes inside the plate if the surface charge density is infinitely large; hence the electrostatic potential is constant there. In this respect, a strongly charged plate behaves like a conductor plate. (4) If Δz is smaller than a Gouy-Chapman length, the correlation potential is dominated by the conventional image charge due to the dielectric discontinuity at the interface. (5) If Δz is larger than a Debye length, the leading order behavior of the correlation potential will depend on the valences of the ions in the electrolyte. Furthermore, inside an asymmetric electrolyte, the correlation potential is singly screened, i.e., it undergoes exponential decay with a decay width equal to the Debye length.

Efficacy of disinfectants containing accelerated hydrogen peroxide against conidial arthrospores and isolated infective spores of Microsporum canis and Trichophyton sp.

PubMed

Moriello, Karen A; Hondzo, Hanna

2014-06-01

Accelerated hydrogen peroxide is a proprietary disinfectant formulation that is available for both commercial and home use and is labelled as antifungal. To determine the antifungal efficacy of accelerated hydrogen peroxide disinfectants against Microsporum and Trichophyton spp. Three products formulated as ready to use and three concentrates were used. Concentrates were tested at dilutions of 1:8, 1:16 (recommended dilution) and 1:32. One product was a surgical instrument disinfectant. Sterile water, sodium hypochlorite (1:32 dilution) and over-the-counter 3% hydrogen peroxide were used as controls. Conidial suspensions contained ~9.6 × 10(5) /mL Microsporum canis, ~1.0 × 10(7) /mL M. gypseum or ~2.0 × 10(7) /mL Trichophyton sp. and were tested at 1:10 dilution. Isolated infective spore suspensions of M. canis from an untreated cat and T. erinacei from an untreated hedgehog were tested at 1:10, 1:5 and 1:1 spore-to-disinfectant dilutions. Too many colonies to count were present on untreated control plates. Accelerated hydrogen peroxide and household hydrogen peroxide inhibited growth of both pathogens in conidial (1:10 dilution) and spore suspensions (1:10, 1:5 and 1:10 dilution). There was no lack of efficacy of products that were >12 months old. Accelerated hydrogen peroxide products are an option for environmental disinfection of surfaces exposed to M. canis and Trichophyton sp. after appropriate gross decontamination and mechanical cleaning with a detergent. The results from conidial testing were identical to those of isolated infected spore testing, which suggests that accelerated hydrogen peroxide products with label claim as antifungal against Trichophyton mentagrophytes may be suitable as an alternative disinfectant to sodium hypochlorite. © 2014 ESVD and ACVD.

Improving Joint Function Using Photochemical Hydrogels for Articular Surface Repair

DTIC Science & Technology

2017-02-01

dilution groups , tdeg increased with increasing concentration of EDC/NHS. Mechanical testing Values for storage modulus in spontaneous control gels (25.86...red) in 48-well nontreated tissue culture plates. As a positive control , a subset group of gels without tethered growth factor was exposed to 0.3 nM...in a cartilage ring and capped with fibrin and collagen gel. A control group consisted of chondrocytes encapsulated in fibrin gel. Constructs were

Microcoupon Assay Of Adhesion And Growth Of Bacterial Films

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Koenig, David W.

1994-01-01

Microbiological assay technique facilitates determination of some characteristics of sessile bacteria like those that attach to and coat interior walls of water-purification systems. Biofilms cause sickness and interfere with purification process. Technique enables direct measurement of rate of attachment of bacterial cells, their metabolism, and effects of chemicals on them. Used to quantify effects of both bactericides and growth-stimulating agents and in place of older standard plate-count and tube-dilution techniques.

Draft Genome Sequences of Two Janthinobacteriumlividum Strains, Isolated from Pristine Groundwater Collected from the Oak Ridge Field Research Center.

PubMed

Wu, Xiaoqin; Deutschbauer, Adam M; Kazakov, Alexey E; Wetmore, Kelly M; Cwick, Bryson A; Walker, Robert M; Novichkov, Pavel S; Arkin, Adam P; Chakraborty, Romy

2017-06-29

We present here the draft genome sequences of two Janthinobacterium lividum strains, GW456P and GW458P, isolated from groundwater samples collected from a background site at the Oak Ridge Field Research Center. Production of a purple pigment by these two strains was observed when grown on diluted (1/10) LB agar plates. Copyright © 2017 Wu et al.

Microbiological Evaluation of Containment Isolators for the Care of Patients with Exotic Diseases.

DTIC Science & Technology

1980-02-01

The microbiological integrity of containment isolators obtained from Vickers Limited Medical Engineering was evaluated using aerosols of Tl coliphage ...diluted in sterile distilled water (SDW). Aliquots for plate counts were mixed with log phase host cells and the mixture spread evenly on pre-dried...of water was diffi- cult to maintain and measure accurately even with sensitive meters, and finally visual observation of the curvature of the side

Bacillus Collagen Like Protein of Anthracis: Immunological and Functional Analyses

DTIC Science & Technology

2007-09-21

heated at 65°C for 30 minutes, diluted, and plated on trypticase soy agar (TSA) to obtain viable counts. Since heat treatment kills the vegetative...purification of that protein by nickel-affinity chromatography are also described in detail elsewhere (Brahmbhatt T.N, lK. Janes, E.S. Stibitz, S.C...Trap Nickel affinity column chromatography with the Fast Phase Liquid Chromatography (FPLC) AKTA system (GE Healthcare, Piscataway, NJ). Rabbit anti

Comparison of the Efficacy of Three Different Mouthrinse Solutions in Decreasing the Level of Streptococcus Mutans in Saliva

PubMed Central

Kocak, Mustafa Murat; Ozcan, Suat; Kocak, Sibel; Topuz, Ozgur; Erten, Hulya

2009-01-01

Objectives The aim of this study was to evaluate the effectiveness of three different antiseptic mouthrinse solutions on the saliva samples obtained from the individuals, who had high caries activity rate. Methods The efficacy of three antiseptic mouthrinses were evaluated in a study with healthy volunteers. The three antiseptic solutions used in this study were 0.1% octenidine dihydrochloride (Octenisept, Schülke&Mayr, UK), 0.12% chlorhexidine digluconate (Kloroben, Drogsan, Turkey) and an antimicrobial enzymatic rinse (Biotene, Laclede, Inc, USA). A total of 27 adult volunteer subjects were participated in the study. The subjects were stratified into three balanced group. Then the mouth rinses were used by each group according to the manufacturer’s directions. The subjects were restricted for 60 minutes for food intake after using the prescribed mouthrinse. The saliva samples were collected from the volunteers at 1, 10 and 60 minutes after their usage in tubes. The tubes were kept in +4°C in a fridge till the evaluation. 10−3 and 10−5 dilutions were prepared for each solution and S. mutans were evaluated according to total number of colony forming unit (CFU) per ml. The dilutions were spreaded on the surface of Brucella agar plates for anaerobic incubation for 48 hours. The dilutions were 100, 10−3 and 10−5 of the solutions Kloroben, Biotene, Octenisept, and the time factor were 0, 1, 10 and 60 minutes. The statistical analyses were performed by Duncan and Bonferroni tests. Results Octenisept was found to be more effective over S. mutans than the other mouthrinse solutions (P<.05). Conclusions All mouthrinse solutions except Biotene were effective on oral microorganisms. PMID:19262732

Verification of an Automated, Digital Dispensing Platform for At-Will Broth Microdilution-Based Antimicrobial Susceptibility Testing.

PubMed

Smith, Kenneth P; Kirby, James E

2016-09-01

With rapid emergence of multidrug-resistant bacteria, there is often a need to perform susceptibility testing for less commonly used or newer antimicrobial agents. Such testing can often be performed only by using labor-intensive, manual dilution methods and lies outside the capacity of most clinical labs, necessitating reference laboratory testing and thereby delaying the availability of susceptibility data. To address the compelling clinical need for microbiology laboratories to perform such testing in-house, we explored a novel, automated, at-will broth microdilution-based susceptibility testing platform. Specifically, we used the modified inkjet printer technology in the HP D300 digital dispensing system to dispense, directly from stock solutions into a 384-well plate, the 2-fold serial dilution series required for broth microdilution testing. This technology was combined with automated absorbance readings and data analysis to determine MICs. Performance was verified by testing members of the Enterobacteriaceae for susceptibility to ampicillin, cefazolin, ciprofloxacin, colistin, gentamicin, meropenem, and tetracycline in comparison to the results obtained with a broth microdilution reference standard. In precision studies, essential and categorical agreement levels were 96.8% and 98.3%, respectively. Furthermore, significantly fewer D300-based measurements were outside ±1 dilution from the modal MIC, suggesting enhanced reproducibility. In accuracy studies performed using a panel of 80 curated clinical isolates, rates of essential and categorical agreement and very major, major, and minor errors were 94%, 96.6%, 0%, 0%, and 3.4%, respectively. Based on these promising initial results, it is anticipated that the D300-based methodology will enable hospital-based clinical microbiology laboratories to perform at-will broth microdilution testing of antimicrobials and to address a critical testing gap. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Active heat exchange system development for latent heat thermal energy storage

NASA Technical Reports Server (NTRS)

Lefrois, R. T.

1980-01-01

Alternative mechanizations of active heat exchange concepts were analyzed for use with heat of fusion Phase Change Materials (PCM's) in the temperature range of 250 C to 350 C for solar and conventional power plant applications. Over 24 heat exchange concepts were reviewed, and eight were selected for detailed assessment. Two candidates were chosen for small-scale experimentation: a coated tube and shell that exchanger, and a direct contact reflux boiler. A dilute eutectic mixture of sodium nitrate and sodium hydroxide was selected as the PCM from over fifty inorganic salt mixtures investigated. Preliminary experiments with various tube coatings indicated that a nickel or chrome plating of Teflon or Ryton coating had promise of being successful. An electroless nickel plating was selected for further testing. A series of tests with nickel-plated heat transfer tubes showed that the solidifying sodium nitrate adhered to the tubes and the experiment failed to meet the required discharge heat transfer rate of 10 kW(t). Testing of the reflux boiler is under way.

Active heat exchange system development for latent heat thermal energy storage

NASA Astrophysics Data System (ADS)

Lefrois, R. T.

1980-03-01

Alternative mechanizations of active heat exchange concepts were analyzed for use with heat of fusion Phase Change Materials (PCM's) in the temperature range of 250 C to 350 C for solar and conventional power plant applications. Over 24 heat exchange concepts were reviewed, and eight were selected for detailed assessment. Two candidates were chosen for small-scale experimentation: a coated tube and shell that exchanger, and a direct contact reflux boiler. A dilute eutectic mixture of sodium nitrate and sodium hydroxide was selected as the PCM from over fifty inorganic salt mixtures investigated. Preliminary experiments with various tube coatings indicated that a nickel or chrome plating of Teflon or Ryton coating had promise of being successful. An electroless nickel plating was selected for further testing. A series of tests with nickel-plated heat transfer tubes showed that the solidifying sodium nitrate adhered to the tubes and the experiment failed to meet the required discharge heat transfer rate of 10 kW(t). Testing of the reflux boiler is under way.

Kinetics of phase separation and coarsening in dilute surfactant pentaethylene glycol monododecyl ether solutions

NASA Astrophysics Data System (ADS)

Tanaka, S.; Kubo, Y.; Yokoyama, Y.; Toda, A.; Taguchi, K.; Kajioka, H.

2011-12-01

We investigated the phase separation phenomena in dilute surfactant pentaethylene glycol monodedecyl ether (C12E5) solutions focusing on the growth law of separated domains. The solutions confined between two glass plates were found to exhibit the phase inversion, characteristic of the viscoelastic phase separation; the majority phase (water-rich phase) nucleated as droplets and the minority phase (micelle-rich phase) formed a network temporarily, then they collapsed into an usual sea-island pattern where minority phase formed islands. We found from the real-space microscopic imaging that the dynamic scaling hypothesis did not hold throughout the coarsening process. The power law growth of the domains with the exponent close to 1/3 was observed even though the coarsening was induced mainly by hydrodynamic flow, which was explained by Darcy's law of laminar flow.

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.

Activity of Imipenem against Klebsiella pneumoniae Biofilms In Vitro and In Vivo

DTIC Science & Technology

2014-02-01

the channels were stained with Live/ Dead BacLight (Invitrogen) for 30 min to determine the live/dead status of bacteria in the remaining biofilms. Due...medium (Fig. 1). To quantify the killing effect of imipenem, we used the mini- mum biofilm eradication concentration (MBEC) assay (11) to determine the...each peg, treated and nontreated, were recovered by sonication, and the number of viable bacteria was determined by serial dilutions and plating. As

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

Numerical analyses of baseline JT-60SA design concepts with the COREDIV code

NASA Astrophysics Data System (ADS)

Zagórski, R.; Gałązka, K.; Ivanova-Stanik, I.; Stępniewski, W.; Garzotti, L.; Giruzzi, G.; Neu, R.; Romanelli, M.

2017-06-01

JT-60SA reference design scenarios at high (#3) and low (#2) density have been analyzed with the help of the self-consistent COREDIV code. Simulations results for a standard C wall and full W wall have been compared in terms of the influence of impurities, both intrinsic (C, W) and seeded (N, Ar, Ne, Kr), on the radiation losses and plasma parameters. For scenario #3 in a C environment, the regime of detachment on divertor plates can be achieved with N or Ne seeding, whereas for the low density and high power scenario (#2), the C and seeding impurity radiation does not effectively reduce power to the targets. In this case, only an increase of either average density or edge density together with Kr seeding might help to develop conditions with strong radiation losses and semi-detached conditions in the divertor. The calculations show that, in the case of a W divertor, the power load to the plate is mitigated by seeding and the central plasma dilution is smaller compared to the C divertor. For the high density case (#3) with Ne seeding, operation in full detachment mode is predicted. Ar seems to be an optimal choice for the low-density high-power scenario #2, showing a wide operating window, whereas Ne leads to high plasma dilution at high seeding levels albeit not achieving semi-detached conditions in the divertor.

Reduction of Salmonella in ground chicken using a bacteriophage.

PubMed

Grant, Ar'Quette; Parveen, Salina; Schwarz, Jurgen; Hashem, Fawzy; Vimini, Bob

2017-08-01

This study's goal was to ascertain the effectiveness of a commercially available Salmonella bacteriophage during ground chicken production focusing on: water source, different Salmonella serovars, and time. Salmonella-free boneless, skinless chicken meat was inoculated with 4.0 Log CFU/cm2 of either a cocktail of 3 Salmonella isolates derived from ground chicken (GC) or a cocktail of 3 Salmonella strains not isolated from ground chicken (non-GC). Bacteriophages were spread onto the chicken using sterile tap or filtered water for 30 min or 8 h. Salmonella was recovered using standard plating method. Greater Salmonella reduction was observed when the bacteriophage was diluted in sterile tap water than in sterile filtered water: 0.39 Log CFU/cm2 and 0.23 Log CFU/cm2 reduction after 30 min, respectively (P < 0.05). The non-GC isolates showed reductions of 0.71 Log CFU/cm2 and 0.90 Log CFU/cm2 after 30 min and 8 h, respectively (P < 0.05). The GC isolates were less sensitive to the bacteriophage: 0.39 Log CFU/cm2 and 0.67 Log CFU/cm2 reductions after 30 min and 8 h, respectively (P < 0.05). In conclusion, bacteriophage reduction was dependent on water used to dilute the bacteriophage, Salmonella's susceptibility to the bacteriophage, and treatment time. © 2017 Poultry Science Association Inc.

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

NASA Technical Reports Server (NTRS)

Schanche, Bradley

2012-01-01

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred µl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

A compact cryogen-free platform operating at 1 K or 50 mK

NASA Astrophysics Data System (ADS)

Matthews, A. J.; Patton, M.; Marsh, T.; van der Vliet, H.

2018-03-01

We report the design and performance characteristics of a compact cryogen-free platform. The system is based around a continuous 1 K pot which operates using a small (10 m3 h‑1) room temperature circulation pump. The pot cools an experimental plate to ≈ 1.2 K, and has a cooling capacity of 100 mW at a temperature ≈ 1.9 K. Cooling the pot from room temperature to < 2 K takes around 12 hours. The temperature range of the platform can be lowered to < 50 mK with the addition of a small dilution refrigerator, using the 1 K pot as a pre-cooling stage for the circulating 3He. The dilution stage has a typical (continuous) cooling capacity of 30 µW at 100 mK (300 µW at 250 mK) and is designed to operate with just 3 litres of (NTP) 3He.

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.

Comparative Evaluation of Antimicrobial Activity of Pomegranate-Containing Mouthwash Against Oral-Biofilm Forming Organisms: An Invitro Microbial Study

PubMed Central

Dabholkar, Charuta Sadanand; Shah, Mona; Bajaj, Monika; Doshi, Yogesh

2016-01-01

Introduction Pomegranate is considered “A pharmacy unto itself”. Hydrolysable tannins called punicalagins which have free scavenging properties are the most abundant polyphenols found in pomegranate-containing mouthwash. Aim To evaluate antimicrobial effect of pomegranate- containing mouthwash on oral biofilm-forming bacteria. Materials and Methods The mouthwashes used were divided into three groups- Group A: Chlorhexidine mouthwash (Hexidine); Group B: Herbal Mouthwash (Hiora) and Group C: Pomegranate-containing Mouthwash (Life-extension). Each mouthwash was diluted to five different concentrations. Reference strains of Streptococcus mutans (S.mutans) (ATCC 25175), Streptococcus salivarius (S.salivarius) (ATCC 7073), and Aggregatibacter actinomycetemcomitans (A.a) (NCTC 9710) were selected as being colonizers in dental biofilm formation. On each culture plate, five wells of 5mm were prepared and mouthwashes with different concentrations were added, followed by incubation in a CO2 jar for 24 hours at 37°C. Inhibition zone diameters were measured using a digital caliper. Results Chlorhexidine (0.12%) presented a zone of inhibition between 38.46% to 96.15% for all the three organisms, while Hiora presented zone of inhibition ranging from 33.33% to 69.23% but was resistant at <10 ml of dilution. Pomegranate mouthwash presented a zone of inhibition ranging from 38.48 to 57.69%, but was resistant at <10ml for S.mutans, and <25ml for A.a and S.salivarius. ANOVA test was done to compare the dilution of mouthwashes for a particular organism and Tukey’s multiple comparison tests were done to find the exact difference. A significant difference was seen between all the three groups at 50ml and 75 ml of dilution. At 75 ml concentration, a statistical difference was found between Groups B & C and Groups A & B; and at 50 ml between Groups A&C. Conclusion All the three types of mouthwash exhibit anti-microbial activity against biofilm forming organisms but at varying concentrations. Although Chlorhexidine still continues to be the gold standard, pomegranate-containing or herbal mouthwashes can be easily substituted for long term use, avoiding the side effects of chlorhexidine. PMID:27135005

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.

Surface Enhanced Raman Spectroscopy for the Rapid Detection and Identification of Microbial Pathogens in Human Serum

DTIC Science & Technology

2014-12-11

and 1 mm depth. Bacterial culture and cell count determination Bacterial species of Acinetobacter baumannii (A. baumannii, ST-3), Escherichia coli...remove all broth components followed by a final resuspension of the pellet in ddH2O back to 1 OD. Cell count was determined by plating the 10 4 , 10 3...10 2 and 10 1 cell dilutions on TSB Nutrient Agar media. Colony forming units (CFU) were counted the following day to confirm bacterial species

Persistence of Antibiotic Resistance Plasmids in Biofilms

DTIC Science & Technology

2013-10-01

Gut )of)louse ATCC no no moderate + TBD,)NR Escherichia+coli ATCC)8739 Complete 4.75 0 NR Feces ATCC no no poor + M3... disinfection  before  the  start  of  the  experiments.  The  routine  procedure   of   running   diluted   bleach   through...overnight   disinfection ,  followed  by  an   additional  2  liters  the  following  day.       • We  plate  and

High throughput analysis of red wine and grape phenolics-adaptation and validation of methyl cellulose precipitable tannin assay and modified Somers color assay to a rapid 96 well plate format.

PubMed

Mercurio, Meagan D; Dambergs, Robert G; Herderich, Markus J; Smith, Paul A

2007-06-13

The methyl cellulose precipitable (MCP) tannin assay and a modified version of the Somers and Evans color assay were adapted to high-throughput (HTP) analysis. To improve efficiency of the MCP tannin assay, a miniaturized 1 mL format and a HTP format using 96 well plates were developed. The Somers color assay was modified to allow the standardization of pH and ethanol concentrations of wine samples in a simple one-step dilution with a buffer solution, thus removing inconsistencies between wine matrices prior to analysis and allowing for its adaptation to a HTP format. Validation studies showed that all new formats were efficient, and results were reproducible and analogous to the original formats.

The prevalence and epidemiology of plasmid-mediated penicillin and tetracycline resistance among Neisseria gonorrhoeae isolates in Guangzhou, China, 2002-2012.

PubMed

Zheng, Heping; Wu, Xingzhong; Huang, Jinmei; Qin, Xiaolin; Xue, Yaohua; Zeng, Weiying; Lan, Yinyuan; Ou, Jiangli; Tang, Sanmei; Fang, Mingheng

2015-10-09

Gonococcal antimicrobial resistance is a global problem. Different resistance plasmids have emerged and spread among the isolates of Neisseria gonorrhoeae worldwide and in China. We conducted this study to monitor the plasmid-mediated penicillin and tetracycline resistance among N. gonorrhoeae isolates in Guangzhou from 2002 to 2012. Consecutive isolates of N. gonorrhoeae were collected from outpatients with gonorrhea attending the STD clinic in Guangdong Provincial Centre for Skin Diseases and STIs Control and Prevention. Penicillinase-producing N. gonorrhoeae (PPNG) isolates were analyzed by the paper acidometric method. Plasmid-mediated resistance to tetracycline in N. gonorrhoeae (TRNG) isolates was screened by the agar plate dilution method. Plasmid types were determined for TRNG and PPNG isolates using polymerase chain reaction (PCR). Minimum inhibitory concentrations (MICs) to penicillin and tetracycline were detected by the agar plate dilution. Of 1378 consecutive N. gonorrhoeae isolates, 429 PPNG and 639 TRNG isolates were identified. The prevalence of PPNG, TRNG, and PPNG/TRNG increased from 18.3 to 47.1 % (χ (2) = 31.57, p < 0.001), from 29.4 to 52.1 % (χ (2) = 16.28, p < 0.001) and from 10.0 to 26.2 % (χ (2) = 10.46, p < 0.001) between 2002 and 2012, respectively. Genotyping of plasmids among PPNGs showed that the majority (93.7 %) of the isolates were the Asian type plasmids, while the African type plasmid emerged in 2008 and rapidly increased to 14.0 % in 2012 (χ (2) = 25.03, p < 0.001). For TRNGs, all 639 isolates carried the Dutch type plasmid. MICs of penicillin G and tetracycline persisted at high levels and the MIC90s were 32-fold higher than the resistant cutoff point over 11 years. The prevalence rates of penicillin- and tetracycline-resistant N. gonorrhoeae varied from 90.9 to 91.1 % and from 88.3 to 89.3 % during 2002 to 2012, respectively. Resistance to penicillin and tetracycline among N. gonorrhoeae isolates remained at high levels in Guangzhou. The Asian type PPNG continued to spread and Dutch type TRNG was still the dominant strain. The African type PPNG has emerged and is spreading rapidly.

Corrosion protection products as a source of bisphenol A and toxicity to the aquatic environment.

PubMed

Vermeirssen, Etiënne L M; Dietschweiler, Conrad; Werner, Inge; Burkhardt, Michael

2017-10-15

Steel components are typically treated with anti-corrosion coatings like epoxy or polyurethane resins to protect the integrity and functioning of steel. Such resins may contain substances, such as bisphenol A (BPA), that have caused concern in a human and environmental toxicological context. We investigated the release of toxicity from four anti-corrosion coatings used in hydraulic and civil engineering. Resins were applied onto glass plates and leachate samples produced by horizontally shaking the plates in water for 7 days. Two experiments were conducted, one with a 1 day and one with a 7 day curing period. Using a suite of bioassays, we tested samples for: agonistic and antagonistic effects on various mammalian nuclear receptors; inhibition of photosynthesis and growth in algae; inhibition of bacterial bioluminescence; and inhibition of water flea reproduction. Concentrations of BPA, bisphenol F and various BPA transformation products were determined by chemical analysis (LC-MS/MS). Bioassay results were evaluated using a scheme developed by DIBt (Centre of Competence for Construction, Berlin, Germany). Three products induced responses in one or more of the measured endpoints and toxicity profiles varied markedly in intensity across products. One product released high amounts of BPA which was associated with effects on nuclear receptor transactivation, requiring a more than 700-fold dilution for effect induction to fall below 20%. The same product was also the most toxic to water flea reproduction, requiring ca. 70-fold dilution for effects to fall below 20%. Another product was highly toxic in terms of bacterial bioluminescence, particularly after a shorter curing time, requiring a ca. 1'300-fold dilution for effects to fall below 20%. The third product required a 22-fold dilution for inhibition of water flea reproduction to drop below 20%. Results show that anti-corrosion coatings based on epoxy resins can be a source of toxicity to the aquatic environment. The fact that some products are more toxic than others highlights opportunities for the development of low risk formulations and products with better environmental performance. Finally, the DIBt scheme provides a useful starting point to develop further ecotoxicity guidelines for testing and data evaluation of leachates from construction materials. Copyright © 2017 Elsevier Ltd. All rights reserved.

Automated reading of MIC microdilution trays containing fluorogenic enzyme substrates with the Sensititre Autoreader.

PubMed Central

Staneck, J L; Allen, S D; Harris, E E; Tilton, R C

1985-01-01

The Sensititre Autoreader is a microcomputer-driven instrument capable of automatically reading antimicrobial susceptibility microdilution trays. The instrument measures the fluorescence liberated by bacterial enzymatic activity on fluorogenic substrates as an indicator of growth in each well. A mathematical algorithm converts the fluorescent signals from an antimicrobial dilution series to an MIC endpoint. A three-center study evaluated the performance of the Autoreader in comparison with MIC determined visually in a duplicate set of control plates lacking fluorogenic substrate. Among 828 isolates of gram-negative bacilli tested against 17 antimicrobial agents, Autoreader 18-h MIC were within +/- 1 twofold dilution of control MIC values (agreement) in 95.3% of instances. In 3.5% of the instances, Autoreader values occurred +/- 2 half-step dilutions from control values (minor discrepancy), and in only 1.2% of instances did Autoreader values deviate from control values by greater than +/- 2 dilution steps (major discrepancy). Agreement, minor discrepancies, and major discrepancies were noted among 148 gram-positive cocci tested against 11 antimicrobial agents in 93.5, 4.8, and 1.7% of the instances, respectively. Over half of the major discrepancies noted with gram-negative bacilli occurred with Proteus mirabilis-beta-lactam combinations, a problem that was resolved when a lower initial inoculum was used. Inter-and intralaboratory reproducibility was excellent. Standard Sensititre susceptibility trays may be instrument read at 18 h reproducibly and accurately with only slight modification of conventional procedures to include fluorogenic enzyme substrates in the incubation broth. PMID:4031033

The antibacterial properties of Malaysian tualang honey against wound and enteric microorganisms in comparison to manuka honey

PubMed Central

Tan, Hern Tze; Rahman, Rosliza Abdul; Gan, Siew Hua; Halim, Ahmad Sukari; Hassan, Siti Asma'; Sulaiman, Siti Amrah; BS, Kirnpal-Kaur

2009-01-01

Background Antibiotic resistance of bacteria is on the rise, thus the discovery of alternative therapeutic agents is urgently needed. Honey possesses therapeutic potential, including wound healing properties and antimicrobial activity. Although the antimicrobial activity of honey has been effectively established against an extensive spectrum of microorganisms, it differs depending on the type of honey. To date, no extensive studies of the antibacterial properties of tualang (Koompassia excelsa) honey on wound and enteric microorganisms have been conducted. The objectives of this study were to conduct such studies and to compare the antibacterial activity of tualang honey with that of manuka honey. Methods Using a broth dilution method, the antibacterial activity of tualang honey against 13 wound and enteric microorganisms was determined; manuka honey was used as the control. Different concentrations of honey [6.25-25% (w/v)] were tested against each type of microorganism. Briefly, two-fold dilutions of honey solutions were tested to determine the minimum inhibitory concentration (MIC) against each type of microorganism, followed by more assays within a narrower dilution range to obtain more precise MIC values. MICs were determined by both visual inspection and spectrophotometric assay at 620 nm. Minimum bactericidal concentration (MBC) also was determined by culturing on blood agar plates. Results By visual inspection, the MICs of tualang honey ranged from 8.75% to 25% compared to manuka honey (8.75-20%). Spectrophotometric readings of at least 95% inhibition yielded MIC values ranging between 10% and 25% for both types of honey. The lowest MBC for tualang honey was 20%, whereas that for manuka honey was 11.25% for the microorganisms tested. The lowest MIC value (8.75%) for both types of honey was against Stenotrophomonas maltophilia. Tualang honey had a lower MIC (11.25%) against Acinetobacter baumannii compared to manuka honey (12.5%). Conclusion Tualang honey exhibited variable activities against different microorganisms, but they were within the same range as those for manuka honey. This result suggests that tualang honey could potentially be used as an alternative therapeutic agent against certain microorganisms, particularly A. baumannii and S. maltophilia. PMID:19754926

Isolation of Phages for Phage Therapy: A Comparison of Spot Tests and Efficiency of Plating Analyses for Determination of Host Range and Efficacy

PubMed Central

Khan Mirzaei, Mohammadali; Nilsson, Anders S.

2015-01-01

Phage therapy, treating bacterial infections with bacteriophages, could be a future alternative to antibiotic treatment of bacterial infections. There are, however, several problems to be solved, mainly associated to the biology of phages, the interaction between phages and their bacterial hosts, but also to the vast variation of pathogenic bacteria which implies that large numbers of different phages are going to be needed. All of these phages must under present regulation of medical products undergo extensive clinical testing before they can be applied. It will consequently be of great economic importance that effective and versatile phages are selected and collected into phage libraries, i.e., the selection must be carried out in a way that it results in highly virulent phages with broad host ranges. We have isolated phages using the Escherichia coli reference (ECOR) collection and compared two methods, spot testing and efficiency of plating (EOP), which are frequently used to identify phages suitable for phage therapy. The analyses of the differences between the two methods show that spot tests often overestimate both the overall virulence and the host range and that the results are not correlated to the results of EOP assays. The conclusion is that single dilution spot tests cannot be used for identification and selection of phages to a phage library and should be replaced by EOP assays. The difference between the two methods can be caused by many factors. We have analysed if the differences and lack of correlation could be caused by lysis from without, bacteriocins in the phage lysate, or by the presence of prophages harbouring genes coding for phage resistance systems in the genomes of the bacteria in the ECOR collection. PMID:25761060

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.

Antibacterial Effect of Azadirachta indica (Neem) or Curcuma longa (Turmeric) against Enterococcus faecalis Compared with That of 5% Sodium Hypochlorite or 2% Chlorhexidine in vitro.

PubMed

Joy Sinha, Dakshita; D S Nandha, Kanwar; Jaiswal, Natasha; Vasudeva, Agrima; Prabha Tyagi, Shashi; Pratap Singh, Udai

2017-01-01

The purpose of this study was to compare the antibacterial properties of Azadirachta indica (neem) or Curcuma longa (turmeric) against Enterococcus faecalis with those of 5% sodium hypochlorite or 2% chlorhexidine as root canal irrigants in vitro. The activity of neem, chlorhexidine, sodium hypochlorite, or turmeric against E. faecalis was measured on agar plates using the agar diffusion method. The tube dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the irrigants used. Chlorhexidine or neem exhibited the greatest antibacterial activity when used as endodontic irrigants against E. faecalis, followed by sodium hypochlorite. No statistically significant difference was observed between neem, sodium hypochlorite, or chlorhexidine. The MIC of neem was 1: 128, which was similar to that of chlorhexidine. The MBC for each of these irrigants was 1: 16. Neem yielded antibacterial activity equivalent to 2% chlorhexidine or sodium hypochlorite against E. faecalis, suggesting that it offers a promising alternative to the other root canal irrigants tested.

A molecular method to assess bioburden embedded within silicon-based resins used on modern spacecraft materials

NASA Astrophysics Data System (ADS)

Stam, Christina N.; Bruckner, James; Spry, J. Andy; Venkateswaran, Kasthuri; La Duc, Myron T.

2012-07-01

Current assessments of bioburden embedded in spacecraft materials are based on work performed in the Viking era (1970s), and the ability to culture organisms extracted from such materials. To circumvent the limitations of such approaches, DNA-based techniques were evaluated alongside established culturing techniques to determine the recovery and survival of bacterial spores encapsulated in spacecraft-qualified polymer materials. Varying concentrations of Bacillus pumilus SAFR-032 spores were completely embedded in silicone epoxy. An organic dimethylacetamide-based solvent was used to digest the epoxy and spore recovery was evaluated via gyrB-targeted qPCR, direct agar plating, most probably number analysis, and microscopy. Although full-strength solvent was shown to inhibit the germination and/or outgrowth of spores, dilution in excess of 100-fold allowed recovery with no significant decrease in cultivability. Similarly, qPCR (quantitative PCR) detection sensitivities as low as ~103 CFU ml-1 were achieved upon removal of inhibitory substances associated with the epoxy and/or solvent. These detection and enumeration methods show promise for use in assessing the embedded bioburden of spacecraft hardware.

Determination of the complete structure of natural lecithins.

PubMed

Kuksis, A; Marai, L

1967-05-01

A method is described for the separation, identification, and quantitative estimation of the individual molecular species occurring in natural lecithin mixtures. Purified lecithin preparations are converted into diglyceride acetates by enzymic dephosphorylation and acetylation. The diglyceride acetates are separated on the basis of the degree of unsaturation and the molecular geometry by means of chromatography on thin layers of silica gel which are impregnated with silver nitrate. The various acetates thus resolved are separately recovered from the plates and diluted with tridecanoin internal standard; the quantitative distribution of the molecular weights is determined by gas chromatography.Suitable aliquots of the saturated and unsaturated diglyceride acetates are further analyzed for over-all and for positional distribution of fatty acids. The identity and proportions of the various lecithins are deduced by integration and normalization of all the experimental data. Where doubt exists, specific diglyceride acetates are isolated by preparative gas chromatography, and their fatty acid composition is determined. The method is illustrated with data obtained for the mixed lecithins of egg yolk. The general approach is applicable to the determination of the structure of other phospholipids of comparable complexity.

Diversity of bacteria and archaea from a landfill in Chandigarh, India as revealed by culture-dependent and culture-independent molecular approaches.

PubMed

Krishnamurthi, S; Chakrabarti, T

2013-02-01

The bacterial community structure of a municipal landfill in Chandigarh, India was analysed by culture-dependent as well as culture-independent molecular approaches, and archaeal structure by the latter method. Samples were collected in two phases from the surface and a depth of 0.91 m in June, 2004 and from 0.91 m, 1.52 m and 1.68 m in May, 2005. After serial dilutions, samples were plated onto tryptic soy agar (TSA), plate count agar (PCA), tryptic soy broth agar (TSBA) and TSBA100 (TSBA diluted 100 times and solidified with agarose), and incubated aerobically at 30°C. The number of bacteria (CFU) on different media ranged between 9.4×10⁵g⁻¹ (on PCA) and 1.9×10⁷g⁻¹ (on TSA) (wet weight). The numbers of bacteria enumerated from plates incubated anaerobically (anaerobic agar and reinforced clostridial agar) were 2.1×10⁷and 1.7×10⁶g⁻¹, respectively. Of the 468 isolated and purified bacteria (183 in the first phase and 285 in the second phase), 135 were characterised using phenotypic characteristics as well as 16S rRNA gene sequence analysis. It was found that members of the phylum Firmicutes were overwhelmingly predominant (86.6%) in the landfill, followed by Actinobacteria (9.6%) and Proteobacteria (3.7%). Among the Firmicutes, at least 17 species from the single genus Bacillus were the most abundant inhabitants of the landfill. Detailed polyphasic characterisation of many of these isolates led to the discovery of a novel genus Paenisporosarcina (and the species P. quisquiliarum), a novel species of Microbacterium, M. immunditiarum, and reclassification of Sporosarcina macmurdoensis, Pelagibacillus goriensis, Bacillus silvestris, Bacillus insolitus, Bacillus psychrotolerans and Bacillus psychrodurans. Culture-independent analysis of two 16S rRNA gene libraries also revealed that the phylum Firmicutes was the predominant group in this community. The diversity of Archaea was found to be limited mainly to members of two orders: Methanosarcinales and Methanomicrobiales of the phylum Euryarchaeota. When these results were compared to those reported earlier on similar studies, it was found that irrespective of differences in composition of municipal solid waste (especially compostable organic matter and paper) and climate, the members of bacterial and archaeal communities in landfills of many countries remained broadly similar. Copyright © 2012 Elsevier GmbH. All rights reserved.

Enumeration of total aerobic microorganisms in foods by SimPlate Total Plate Count-Color Indicator methods and conventional culture methods: collaborative study.

PubMed

Feldsine, Philip T; Leung, Stephanie C; Lienau, Andrew H; Mui, Linda A; Townsend, David E

2003-01-01

The relative efficacy of the SimPlate Total Plate Count-Color Indicator (TPC-CI) method (SimPlate 35 degrees C) was compared with the AOAC Official Method 966.23 (AOAC 35 degrees C) for enumeration of total aerobic microorganisms in foods. The SimPlate TPC-CI method, incubated at 30 degrees C (SimPlate 30 degrees C), was also compared with the International Organization for Standardization (ISO) 4833 method (ISO 30 degrees C). Six food types were analyzed: ground black pepper, flour, nut meats, frozen hamburger patties, frozen fruits, and fresh vegetables. All foods tested were naturally contaminated. Nineteen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery among the SimPlate methods and their corresponding reference methods. Mean log counts between the 2 reference methods were also very similar. Repeatability (Sr) and reproducibility (SR) standard deviations were similar among the 3 method comparisons. The SimPlate method (35 degrees C) and the AOAC method were comparable for enumerating total aerobic microorganisms in foods. Similarly, the SimPlate method (30 degrees C) was comparable to the ISO method when samples were prepared and incubated according to the ISO method.

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.

The role of long-range forces in the formation of thin liquid films on metals

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

Gyory, J.R.; Muller, R.H.

1987-06-01

White-light multiple beam interference is used to study the drainage of aqueous electrolytes from vertically optically smooth platinum and gold plates. Bulk liquid is in contact with the bottom of the metal plate. For short times following the lowering of the bulk liquid level, the change in the film profile agrees with that expected from viscous drainage. However, at long times, the film profile deviates from that expected and eventually becomes independent of time at a thickness between 0.08 and 0.25 micrometers. These profiles are best represented by a function dependent on the inverse cube root of height. The thicknessmore » of the equilibrium film profiles with increasing electrolyte concentration. A model based on long range van der Waals interactions resulting in a repulsive force between the interfaces of the film is shown to predict the correct profile shape, and for dilute electrolytes, the correct film thickness. This model also predicts increasing film thickness for increasing electrolyte concentration. The strength of this interaction is characterized by the Hamaker constant which can be calculated from the dielectric functions evaluated at imaginary frequencies of the film and substrate. For metals, this function is generated from spectral absorption data, limiting behavior for low and high frequencies, and by use of the Kramers-Kronig transformation. Hamaker constants calculated from the dielectric functions generated in this manner agree well with those derived from film profiles for dilute electrolytes.« less

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...

gcpA (stm1987) is critical for cellulose production and biofilm formation on polystyrene surface by Salmonella enterica serovar Weltevreden in both high and low nutrient medium.

PubMed

Bhowmick, Patit Paban; Devegowda, Devananda; Ruwandeepika, H A Darshanee; Fuchs, Thilo M; Srikumar, Shabarinath; Karunasagar, Iddya; Karunasagar, Indrani

2011-02-01

Biofilm formation by Salmonella is a serious concern in the food-processing industry and the persistence of the organism in biofilms becomes a constant source of contamination. Since there is zero tolerance for Salmonella in foods, it is important to understand the mechanism of biofilm formation and to prevent the formation. Therefore, this study aimed at investigating the biofilm-forming ability of seafood isolates of Salmonella enterica serovar Weltevreden (S. Weltevreden) under two different nutrient conditions (normal strength trypticase soy broth (TSB) and 1:100 diluted TSB). The role of cellulose production in biofilm formation and in the expression of multicellular behavior (rough, dark, red morphotype: rdar) was investigated. Fourteen isolates of seafood associated S. Weltevreden were studied for biofilm production in polystyrene microtitre plates. Only one (SW49) of 14 was a strong biofilm former on polystyrene template and was able to produce biofilm in both undiluted TSB and 1:100 diluted TSB at 24h. All others produced moderate or weak biofilms which was higher in 1:100 diluted TSB compared to undiluted medium. All the isolates except one were positive by PCR for the three genes, gcpA (stm1987), adrA (yaiC) and csgD. Gene expression of gcpA, adrA and csgD was studied by real-time PCR with the one strong (SW49) and one representative weak (SW30) biofilm former. In SW49 at 24h of incubation, the expression of gcpA from biofilm cells was 33 and 36 times higher than from planktonic cells grown in TSB and diluted TSB respectively and at 72h the expression from biofilm cells was 57 and 61 times higher than that from planktonic cells. Quantification of gene expression did not reveal any significant difference in the expression of csgD and adrA gene. Deletion of gcpA in SW49 resulted in its inability to produce cellulose and consequent inability to bind calcoflour, inability to form rdar colony on Congo Red-agar plates and failure to produce biofilm on polystyrene substrate. The data indicated that, in case of S. Weltevreden, gcpA is critical for activating cellulose synthesis and biofilm formation both in undiluted and diluted TSB. The results of this study suggest the existence of an alternative biofilm regulatory pathway in S. Weltevreden. Role of adrA in cellulose production in nutrient rich medium is known but role of gepA in the above phenomenon is proved in this study. An understanding of the genes involved would help in looking at strategies of repression of the gene to control formation of biofilm. Copyright © 2010 Elsevier Ltd. All rights reserved.

[Difference of three standard curves of real-time reverse-transcriptase PCR in viable Vibrio parahaemolyticus quantification].

PubMed

Jin, Mengtong; Sun, Wenshuo; Li, Qin; Sun, Xiaohong; Pan, Yingjie; Zhao, Yong

2014-04-04

We evaluated the difference of three standard curves in quantifying viable Vibrio parahaemolyticus in samples by real-time reverse-transcriptase PCR (Real-time RT-PCR). The standard curve A was established by 10-fold diluted cDNA. The cDNA was reverse transcripted after RNA synthesized in vitro. The standard curve B and C were established by 10-fold diluted cDNA. The cDNA was synthesized after RNA isolated from Vibrio parahaemolyticus in pure cultures (10(8) CFU/mL) and shrimp samples (10(6) CFU/g) (Standard curve A and C were proposed for the first time). Three standard curves were performed to quantitatively detect V. parahaemolyticus in six samples, respectively (Two pure cultured V. parahaemolyticus samples, two artificially contaminated cooked Litopenaeus vannamei samples and two artificially contaminated Litopenaeus vannamei samples). Then we evaluated the quantitative results of standard curve and the plate counting results and then analysed the differences. The three standard curves all show a strong linear relationship between the fractional cycle number and V. parahaemolyticus concentration (R2 > 0.99); The quantitative results of Real-time PCR were significantly (p < 0.05) lower than the results of plate counting. The relative errors compared with the results of plate counting ranked standard curve A (30.0%) > standard curve C (18.8%) > standard curve B (6.9%); The average differences between standard curve A and standard curve B and C were - 2.25 Lg CFU/mL and - 0.75 Lg CFU/mL, respectively, and the mean relative errors were 48.2% and 15.9%, respectively; The average difference between standard curve B and C was among (1.47 -1.53) Lg CFU/mL and the average relative errors were among 19.0% - 23.8%. Standard curve B could be applied to Real-time RT-PCR when quantify the number of viable microorganisms in samples.

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.

The inhibitory effect of Thymus vulgaris extracts on the planktonic form and biofilm structures of six human pathogenic bacteria

PubMed Central

Mohsenipour, Zeinab; Hassanshahian, Mehdi

2015-01-01

Objective: Microorganisms are responsible for many problems in industry and medicine because of biofilm formation. Therefore, this study was aimed to examine the effect of Thymus vulgaris (T. vulgaris) extracts on the planktonic form and biofilm structures of six pathogenic bacteria. Materials and methods: Antimicrobial activities of the plant extracts against the planktonic form of the bacteria were determined using the disc diffusion method. MIC and MBC values were evaluated using macrobroth dilution technique. Anti-biofilm effects were assessed by microtiter plate method. Results: According to disc diffusion test (MIC and MBC), the ability of Thymus vulgaris (T. vulgaris ) extracts for inhibition of bacteria in planktonic form was confirmed. In dealing with biofilm structures, the inhibitory effect of the extracts was directly correlated to their concentration. Except for the inhibition of biofilm formation, efficacy of each extract was independent from type of solvent. Conclusion: According to the potential of Thymus vulgaris (T. vulgaris) extracts to inhibit the test bacteria in planktonic and biofilm form, it can be suggested that Thymus vulgaris (T. vulgaris) extracts can be applied as antimicrobial agents against the pathogenic bacteria particularly in biofilm forms. PMID:26442753

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.

Antifungal mechanisms supporting boric acid therapy of Candida vaginitis.

PubMed

De Seta, Francesco; Schmidt, Martin; Vu, Bao; Essmann, Michael; Larsen, Bryan

2009-02-01

Boric acid is a commonly cited treatment for recurrent and resistant yeast vaginitis, but data about the extent and mechanism of its antifungal activity are lacking. The aim of this study was to use in vitro methods to understand the spectrum and mechanism of boric acid as a potential treatment for vaginal infection. Yeast and bacterial isolates were tested by agar dilution to determine the intrinsic antimicrobial activity of boric acid. Established microbial physiology methods illuminated the mechanism of the action of boric acid against Candida albicans. C. albicans strains (including fluconazole-resistant strains) were inhibited at concentrations attainable intravaginally; as were bacteria. Broth dilution MICs were between 1563 and 6250 mg/L and boric acid proved fungistatic (also reflected by a decrease in CO(2) generation); prolonged culture at 50,000 mg/L was fungicidal. Several organic acids in yeast nitrogen broth yielded a lower pH than equimolar boric acid and sodium borate but were less inhibitory. Cold or anaerobic incubation protected yeast at high boric acid concentrations. Cells maintained integrity for 6 h in boric acid at 37 degrees C, but after 24 h modest intrusion of propidium iodide occurred; loss of plate count viability preceded uptake of vital stain. Growth at sub-MIC concentrations of boric acid decreased cellular ergosterol. The drug efflux pump CDR1 did not protect Candida as CDR1 expression was abrogated by boric acid. Boric acid interfered with the development of biofilm and hyphal transformation. Boric acid is fungistatic to fungicidal depending on concentration and temperature. Inhibition of oxidative metabolism appears to be a key antifungal mechanism, but inhibition of virulence probably contributes to therapeutic efficacy in vivo.

A multiwell format assay for heparanase.

PubMed

Behzad, Farhad; Brenchley, Paul E C

2003-09-15

This assay employs a biotinylated heparan sulfate glycosaminoglycan (HSGAG) substrate that is covalently linked to the surface of 96-well immunoassay plates. The ratio of biotin:HSGAG and the coating concentration of substrate bound to the wells have been optimized and allow removal of biotin HSGAG within 60 min of incubation at 37 degrees C in assay buffer with a standard dilution of bacterial heparitinase or platelet heparanase. Loss of biotin signal from the well surface is detected on incubation with peroxidase-streptavidin followed by color development using 3,3',5,5'-tetramethylbenzidine as the peroxidase substrate. The new assay allows specific detection of heparanase activity in multiple samples in a total time of 3 h including a 1-h substrate digestion step and is a significant improvement with regard to sensitivity, specificity, and ease of handling of multiple samples compared to other described assays. Heparanase specifically degrades the biotinylated HSGAG substrate, when used with an optimized assay buffer. A range of enzymes including collagenase, trypsin, plasmin, pepsin, chondroitinases, hyaluronidase, and neuraminidase show no effect on the substrate under optimized assay conditions. The covalent linkage of the substrate to the well prevents leaching of substrate and allows preparation and long-term storage of substrate-coated plates. The assay can be used to detect heparanase levels in clinical samples and cell culture supernatants and is ideal as a screening method for antagonists of enzyme activity.

Bioconversion of straw into improved fodder: fungal flora decomposing rice straw.

PubMed

Helal, G A

2005-09-01

The fungal flora decomposing rice straw were investigated all over the soil of Sharkia Province, east of Nile Delta, Egypt, using the nylon net bag technique. Sixty-four straw-decomposing species belonging to 30 genera were isolated by the dilution plate method in ground rice straw-Czapek's agar medium at pH 6. The plates were incubated separately at 5℃, 25℃ and 45℃, respectively. Twenty nine species belonging to 14 genera were isolated at 5℃. The most frequent genus was Penicillium (seven species), and the next frequent genera were Acremonium (three species), Fusarium (three species), Alternaria, Chaetomium, Cladosporium, Mucor, Stachybotrys (two species) and Rhizopus stolonifer. At 25℃, 47 species belonging to 24 genera were isolated. The most frequent genus was Aspergillus (nine species), and the next frequent genera were ranked by Penicillium (five species), Chaetomium (three species), Fusarium (three species). Each of Alternaria, Cladosporium, Mucor, Myrothecium and Trichoderma was represented by two species. At 45℃, 15 species belonging to seven genera were isolated. These were seven species of Aspergillus, two species of Chaetomium and two species of Emericella, while Humicola, Malbranchea, Rhizomucor and Talaromyces were represented by one species respectively. The total counts of fungi the genera, and species per gram of dry straw were significantly affected by incubation temperature and soil analysis (P < 0.05).

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.

The shock sensitivity of nitromethane/methanol mixtures

NASA Astrophysics Data System (ADS)

Bartram, Brian; Dattelbaum, Dana; Sheffield, Steve; Gibson, Lee

2013-06-01

The dilution of liquid explosives has multiple effects on detonation properties including an increase in critical diameter, spatiotemporal lengthening of the chemical reaction zone, and the development of propagating wave instabilities. Earlier detonation studies of NM/methanol mixtures have shown several effects of increasing dilution, including: 1) a continual increase in the critical diameter, 2) lowering of the Chapman-Jouguet detonation pressure, and 3) slowing of the steady detonation velocity (Koldunov et al., Comb. Expl. Shock Waves). Here, we present the results of a series of gas gun-driven plate-impact experiments to study the shock-to-detonation transition in NM/methanol mixtures. Embedded electromagnetic gauges were used to obtain in situ particle velocity wave profiles at multiple Lagrangian positions in the initiating explosive mixture. From the wave profiles obtained in each experiment, an unreacted Hugoniot locus, the initiation mechanism, and the overtake-time-to-detonation were obtained as a function of shock input condition for mixture concentrations from 100% NM to 50 wt%/50 wt% NM/methanol. Desensitization with dilution is less than expected. For example, little change in overtake time occurs in 80 wt%/20 wt% NM/methanol when compared with neat NM. Furthermore, the shock wave profiles from the gauges indicate that wave instabilities grow in as the overdriven detonation wave settles down following the shock-to-detonation transition.

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.

Process and reactor design for biophotolytic hydrogen production.

PubMed

Tamburic, Bojan; Dechatiwongse, Pongsathorn; Zemichael, Fessehaye W; Maitland, Geoffrey C; Hellgardt, Klaus

2013-07-14

The green alga Chlamydomonas reinhardtii has the ability to produce molecular hydrogen (H2), a clean and renewable fuel, through the biophotolysis of water under sulphur-deprived anaerobic conditions. The aim of this study was to advance the development of a practical and scalable biophotolytic H2 production process. Experiments were carried out using a purpose-built flat-plate photobioreactor, designed to facilitate green algal H2 production at the laboratory scale and equipped with a membrane-inlet mass spectrometry system to accurately measure H2 production rates in real time. The nutrient control method of sulphur deprivation was used to achieve spontaneous H2 production following algal growth. Sulphur dilution and sulphur feed techniques were used to extend algal lifetime in order to increase the duration of H2 production. The sulphur dilution technique proved effective at encouraging cyclic H2 production, resulting in alternating Chlamydomonas reinhardtii recovery and H2 production stages. The sulphur feed technique enabled photobioreactor operation in chemostat mode, resulting in a small improvement in H2 production duration. A conceptual design for a large-scale photobioreactor was proposed based on these experimental results. This photobioreactor has the capacity to enable continuous and economical H2 and biomass production using green algae. The success of these complementary approaches demonstrate that engineering advances can lead to improvements in the scalability and affordability of biophotolytic H2 production, giving increased confidence that H2 can fulfil its potential as a sustainable fuel of the future.

Isolation of Raoultella planticola from refillable antimicrobial liquid soap dispensers in a dental setting

PubMed Central

Tomlin, Nancy; Ruby, John D.

2014-01-01

Background Liquid antimicrobial soaps are commonly used in the dental healthcare setting for hand washing to minimize the potential spread of infectious agents to healthcare workers (HCW) and patients. The purpose of the current study was to evaluate possible bacterial contamination of antimicrobial liquid soap dispensers located in two institutional comprehensive dental care clinics. Methods Fourteen soap dispensers and original stock containers were sampled. A 1 ml aliquot was diluted in 10 ml of phosphate buffer (Tween 80). Serial dilutions were plated in duplicate on neutralizing agar and incubated for 7 days. Molecular identification was performed using 500 bp comparisons of 16S rDNA sequencing. Taq PCR was performed with sequence specific primers for Raoultella species. Results Bacterial growth was observed at 18 hours for 57% (8/14) of soap dispenser samples. Bacterial densities ranged from 4 ×102–6 ×109 CFU/mL. Original commercial containers exhibited no growth. Isolates were identified as Raoultella (Klebsiella) planticola. Conclusions This is the first study indicating recovery of R. planticola from antimicrobial liquid soap dispensers. R. planticola is a recognized environmental opportunistic pathogen that potentially poses a health concern. Practical Implications These findings indicate compliance problems with infection prevention recommendations and support the CDC’s recommendation that dispensers should not be “topped off”. High bacterial loads of R. planticola are inconsistent with infection control practices and are a concern since transmission and possible infection to the HCW or the patient may occur. PMID:25819655

In vitro activity of a polyhexanide-betaine solution against high-risk clones of multidrug-resistant nosocomial pathogens.

PubMed

López-Rojas, Rafael; Fernández-Cuenca, Felipe; Serrano-Rocha, Lara; Pascual, Álvaro

2017-01-01

To determine the in vitro activity of a polyhexanide-betaine solution against collection strains and multidrug-resistant (MDR) nosocomial isolates, including high-risk clones. We studied of 8 ATCC and 21 MDR clinical strains of Staphylococcus aureus, Enterococcus faecium, Enterococcus faecalis, Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, including the multiresistant high-risk clones. The MICs and MBCs of a 0.1% polyhexanide-0.1% betaine solution were determined by microdilution. For each species, strains with the highest MICs were selected for further experiments. The dilution-neutralization test (PrEN 12054) was performed by incubating bacterial inocula of 10 6 CFU/mL for 1min with undiluted 0.1% polyhexanide-betaine solution. The CFUs were counted after neutralization. Growth curves and time-kill curves at concentrations of 0.25, 1, 4, and 8×MIC, were performed. MICs of recovered strains were determined when regrowth was observed in time-kill studies after 24h of incubation. Strains with reduced susceptibility were selected by serial passage on plates with increasing concentrations of polyhexanide-betaine, and MICs were determined. Polyhexanide-betaine MIC range was 0.5-8mg/L. MBCs equalled or were 1 dilution higher than MICs. The dilution-neutralization method showed total inoculum clearance of all strains. In time-kill curves, no regrowth was observed at 4×MIC, except for S. aureus (8×MIC). Increased MICs were not observed in time-kill curves, or after serial passages after exposure to polyhexanide-betaine. Polyhexanide-betaine presented bactericidal activity against all MDR clinical isolates tested, including high-risk clones, at significantly lower concentrations and time of activity than those commercially used. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Total testosterone quantitative measurement in serum by LC-MS/MS☆

PubMed Central

Wang, Yuesong; Gay, Gabrielle D.; Botelho, Julianne Cook; Caudill, Samuel P.; Vesper, Hubert W.

2016-01-01

Reliable measurement of total testosterone is essential for the diagnosis, treatment and prevention of a number of hormone-related diseases affecting adults and children. A mass spectrometric method for testosterone determination in human serum was carefully developed and thoroughly validated. Total testosterone from 100 μL serum is released from proteins with acidic buffer and isolated by two serial liquid–liquid extraction steps. The first extraction step isolates the lipid fractions from an acidic buffer solution using ethyl acetate and hexane. The organic phase is dried down and reconstituted in a basic buffer solution. The second extraction step removes the phospholipids and other components by hexane extraction. Liquid chromatography–isotopic dilution tandem mass spectrometry is used to quantify the total testosterone. The sample preparation is automatically conducted in a liquid-handling system with 96-deepwell plates. The method limit of detection is 9.71 pmol/L (0.280 ng/dL) and the method average percent bias is not significantly different from reference methods. The performance of this method has proven to be consistent with the method precision over a 2-year period ranging from 3.7 to 4.8% for quality control pools at the concentrations 0.527, 7.90 and 30.7 nmol/L (15.2, 228, and 886 ng/dL), respectively. This method provides consistently high accuracy and excellent precision for testosterone determination in human serum across all clinical relevant concentrations. PMID:24960363

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.

Thermal conductance measurements of bolted copper joints for SuperCDMS

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

Schmitt, R.; Tatkowski, Greg; Ruschman, M.

2015-09-01

Joint thermal conductance testing has been undertaken for bolted copper to copper connections from 60 mK to 26 K. This testing was performed to validate an initial design basis for the SuperCDMS experiment, where a dilution refrigerator will be coupled to a cryostat via multiple bolted connections. Copper used during testing was either gold plated or passivated with citric acid to prevent surface oxidation. Results obtained are well fit by a power law regression of joint thermal conductance to temperature and match well with data collected during a literature review.

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

R-plasmid transfer in a wastewater treatment plant.

PubMed Central

Mach, P A; Grimes, D J

1982-01-01

Enteric bacteria have been examined for their ability to transfer antibiotic resistance in a wastewater treatment plant. Resistant Salmonella enteritidis, Proteus mirabilis, and Escherichia coli were isolated from clinical specimens and primary sewage effluent. Resistance to ampicillin, chloramphenicol, streptomycin, sulfadiazine, and tetracycline was demonstrated by spread plate and tube dilution techniques. Plasmid mediation of resistance was shown by ethidium bromide curing, agarose gel electrophoresis, and direct cell transfer. Each donor was mated with susceptible E. coli and Shigella sonnei. Mating pairs (and recipient controls) were suspended in unchlorinated primary effluent that had been filtered and autoclaved. Suspensions were added to membrane diffusion chambers which were then placed in the primary and secondary setting tanks of the wastewater treatment plant. Resistant recombinants were detected by replica plating nutrient agar master plates onto xylose lysine desoxycholate agar plates that contained per milliliter of medium 10 micrograms of ampicillin, 30 micrograms of chloramphenicol, 10 micrograms of streptomycin, 100 micrograms of sulfadiazine, or 30 micrograms of tetracycline. Mean transfer frequencies for laboratory matings were 2.1 X 10(-3). In situ matings for primary and secondary settling resulted in frequencies of 4.9 X 10(-5) and 7.5 X 10(-5), respectively. These values suggest that a significant level of resistance transfer occurs in wastewater treatment plants in the absence of antibiotics as selective agents. Images PMID:6760813

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.

Surface topographical changes measured by phase-locked interferometry

NASA Technical Reports Server (NTRS)

Lauer, J. L.; Fung, S. S.

1984-01-01

An electronic optical laser interferometer capable of resolving depth differences of as low as 30 A and planar displacements of 6000 A was constructed to examine surface profiles of bearing surfaces without physical contact. Topological chemical reactivity was determined by applying a drop of dilute alcoholic hydrochloric acid and measuring the profile of the solid surface before and after application of this probe. Scuffed bearing surfaces reacted much faster than virgin ones but that bearing surfaces exposed to lubricants containing an organic chloride reacted much more slowly. The reactivity of stainless steel plates, heated in a nitrogen atmosphere to different temperatures, were examined later at ambient temperature. The change of surface contour as a result of the probe reaction followed Arrhenius-type relation with respect to heat treatment temperature. The contact area of the plate of a ball/plate sliding elastohydrodynamic contact run on trimethylopropane triheptanoate with or without additives was optically profiled periodically. As scuffing was approached, the change of profile within the contact region changed much more rapidly by the acid probe and assumed a constant high value after scuffing. A nonetching metallurgical phase was found in the scuff mark, which was apparently responsible for the high reactivity.

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...

Cell viability of mycorrhiza helper bacteria solid inoculant in different carrier material

NASA Astrophysics Data System (ADS)

Asyiah, Iis Nur; Hindersah, Reginawanti; Harni, Rita

2018-02-01

Roots of food crops are colonized by nonpathogenic mycorrhizal fungi which show natural ability to control plant pathogen. Mycorrhizal establishment in plant roots is affected by rhizobacteria, known as mycorrhiza helper bacteria (MHB), which has synergetic effects on mycorrhizal associations. Laboratory experiment has been conducted to assess the best carrier material to develop well-qualified MHB of Pseudomonas diminuta and Bacillus subtilis solid inoculant. Carrier materials were 100 mesh organic matter of agricultural waste. Different spore concentration of both bacterial liquid inoculants were grown on three kinds of 100-mesh organic matter and stored at room temperature up to 90 days. Cell viability of both MHB were counted by serial dilution plate method by using specific medium. The results showed that sugar cane baggase ash was the best carrier material to maintain cell viability for both MHB. However, the population of Pseudomonas diminuta and Bacillus subtilis in sugar cane baggase ash were slightly decreased after 90 days. The use of sugarcane baggase ash for solid MHB inoculant development could be suggested.

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.

Protocols for dry DNA storage and shipment at room temperature

PubMed Central

Ivanova, Natalia V; Kuzmina, Masha L

2013-01-01

The globalization of DNA barcoding will require core analytical facilities to develop cost-effective, efficient protocols for the shipment and archival storage of DNA extracts and PCR products. We evaluated three dry-state DNA stabilization systems: commercial Biomatrica® DNAstable® plates, home-made trehalose and polyvinyl alcohol (PVA) plates on 96-well panels of insect DNA stored at 56 °C and at room temperature. Controls included unprotected samples that were stored dry at room temperature and at 56 °C, and diluted samples held at 4 °C and at −20 °C. PCR and selective sequencing were performed over a 4-year interval to test the condition of DNA extracts. Biomatrica® provided better protection of DNA at 56 °C and at room temperature than trehalose and PVA, especially for diluted samples. PVA was the second best protectant after Biomatrica® at room temperature, whereas trehalose was the second best protectant at 56 °C. In spite of lower PCR success, the DNA stored at −20 °C yielded longer sequence reads and stronger signal, indicating that temperature is a crucial factor for DNA quality which has to be considered especially for long-term storage. Although it is premature to advocate a transition to DNA storage at room temperature, dry storage provides an additional layer of security for frozen samples, protecting them from degradation in the event of freezer failure. All three forms of DNA preservation enable shipment of dry DNA and PCR products between barcoding facilities. PMID:23789643

A high throughput passive dosing format for the Fish Embryo Acute Toxicity test.

PubMed

Vergauwen, Lucia; Schmidt, Stine N; Stinckens, Evelyn; Maho, Walid; Blust, Ronny; Mayer, Philipp; Covaci, Adrian; Knapen, Dries

2015-11-01

High throughput testing according to the Fish Embryo Acute Toxicity (FET) test (OECD Testing Guideline 236) is usually conducted in well plates. In the case of hydrophobic test substances, sorptive and evaporative losses often result in declining and poorly controlled exposure conditions. Therefore, our objective was to improve exposure conditions in FET tests by evaluating a passive dosing format using silicone O-rings in standard 24-well polystyrene plates. We exposed zebrafish embryos to a series of phenanthrene concentrations until 120h post fertilization (hpf), and obtained a linear dilution series. We report effect values for both mortality and sublethal morphological effects based on (1) measured exposure concentrations, (2) (lipid normalized) body residues and (3) chemical activity. The LC50 for 120hpf was 310μg/L, CBR50 (critical body residue) was 2.72mmol/kg fresh wt and La50 (lethal chemical activity) was 0.047. All values were within ranges expected for baseline toxicity. Impaired swim bladder inflation was the most pronounced morphological effect and swimming activity was reduced in all exposure concentrations. Further analysis showed that the effect on swimming activity was not attributed to impaired swim bladder inflation, but rather to baseline toxicity. We conclude that silicone O-rings (1) produce a linear dilution series of phenanthrene in the 120hpf FET test, (2) generate and maintain aqueous concentrations for reliable determination of effect concentrations, and allow for obtaining mechanistic toxicity information, and (3) cause no toxicity, demonstrating its potential as an extension of the FET test when testing hydrophobic chemicals. Copyright © 2015 Elsevier Ltd. All rights reserved.

Wear studies of all UHMWPE couples under various bio-tribological conditions.

PubMed

Joyce, T J; Unsworth, A

2004-01-01

Wear tests were undertaken in which ultra high molecular weight polyethylene (UHMWPE) was rubbed against itself. Tests primarily employed a pin-on-plate wear test machine, with distilled water, Ringer solution and dilute bovine serum being used as the lubricants. Loads of 10N and 40N were employed, and some test pins had a rotational motion added. In all cases wear was high, with mean wear factors of up to 91 10 -6 mm3/Nm being measured, but the addition of rotation reduced the amount of material worn from the test plates. In the presence of bovine serum and under reciprocation only, pin wear was relatively low. With bovine serum as the lubricant, total mean wear factors for the UHMWPE couples were calculated to be in the range of 35 to 58 10-6mm3/Nm. Therefore the pin-on-plate tests showed that the choice of lubricant as well as the motion applied to the test pin had a significant influence on the wear volumes measured. A two-piece UHMWPE 'prosthesis' with matching hemispherical faces was fabricated and tested on a finger simulator. Distilled water was used as the lubricant and wear factors were found to be greater for the metacarpal component, 21 10 -6mm3/Nm, than the phalangeal component, 3 10-6mm3/Nm, after ten million cycles of testing. This result paralleled the greater wear seen by the plate than by the pin in the pin-on-plate tests under reciprocating motion. (Journal of Applied Biomaterials & Biomechanics 2004; 2: 29-34).

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.

Growth of 1.5 micron gallium indium nitrogen arsenic antimonide vertical cavity surface emitting lasers by molecular beam epitaxy

NASA Astrophysics Data System (ADS)

Wistey, Mark Allan

Fiber optics has revolutionized long distance communication and long haul networks, allowing unimaginable data speeds and noise-free telephone calls around the world for mere pennies per hour at the trunk level. But the high speeds of optical fiber generally do not extend to individual workstations or to the home, in large part because it has been difficult and expensive to produce lasers which emitted light at wavelengths which could take advantage of optical fiber. One of the most promising solutions to this problem is the development of a new class of semiconductors known as dilute nitrides. Dilute nitrides such as GaInNAs can be grown directly on gallium arsenide, which allows well-established processing techniques. More important, gallium arsenide allows the growth of vertical-cavity surface-emitting lasers (VCSELs), which can be grown in dense, 2D arrays on each wafer, providing tremendous economies of scale for manufacturing, testing, and packaging. Unfortunately, GaInNAs lasers have suffered from what has been dubbed the "nitrogen penalty," with high thresholds and low efficiency as the fraction of nitrogen in the semiconductor was increased. This thesis describes the steps taken to identify and essentially eliminate the nitrogen penalty. Protecting the wafer surface from plasma ignition, using an arsenic cap, greatly improved material quality. Using a Langmuir probe, we further found that the nitrogen plasma source produced a large number of ions which damaged the wafer during growth. The ions were dramatically reduced using deflection plates. Low voltage deflection plates were found to be preferable to high voltages, and simulations showed low voltages to be adequate for ion removal. The long wavelengths from dilute nitrides can be partly explained by wafer damage during growth. As a result of these studies, we demonstrated the first CW, room temperature lasers at wavelengths beyond 1.5mum on gallium arsenide, and the first GaInNAs(Sb) VCSELs beyond 1.31mum: 1.46mum. These techniques offer the promise of inexpensive, high speed fiber networking.

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)

Sub-Plate Overlap Code Documentation

NASA Technical Reports Server (NTRS)

Taff, L. G.; Bucciarelli, B.; Zarate, N.

1997-01-01

An expansion of the plate overlap method of astrometric data reduction to a single plate has been proposed and successfully tested. Each plate is (artificially) divided into sub-plates which can then be overlapped. This reduces the area of a 'plate' over which a plate model needs to accurately represent the relationship between measured coordinates and standard coordinates. Application is made to non-astrographic plates such as Schmidt plates and to wide-field astrographic plates. Indeed, the method is completely general and can be applied to any type of recording media.

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.

A Study on the Saving Method of Plate Jigs in Hull Block Butt Welding

NASA Astrophysics Data System (ADS)

Ko, Dae-Eun

2017-11-01

A large amount of plate jigs is used for alignment of welding line and control of welding deformations in hull block assembly stage. Besides material cost, the huge working man-hours required for working process of plate jigs is one of the obstacles in productivity growth of shipyard. In this study, analysis method was proposed to simulate the welding deformations of block butt joint with plate jigs setting. Using the proposed analysis method, an example simulation was performed for actual panel block joint to investigate the saving method of plate jigs. Results show that it is possible to achieve two objectives of quality accuracy of the hull block and saving the plate jig usage at the same time by deploying the plate jigs at the right places. And the proposed analysis method can be used in establishing guidelines for the proper use of plate jigs in block assembly stage.

Elastostatic stress analysis of orthotropic rectangular center-cracked plates

NASA Technical Reports Server (NTRS)

Gyekenyesi, G. S.; Mendelson, A.

1972-01-01

A mapping-collocation method was developed for the elastostatic stress analysis of finite, anisotropic plates with centrally located traction-free cracks. The method essentially consists of mapping the crack into the unit circle and satisfying the crack boundary conditions exactly with the help of Muskhelishvili's function extension concept. The conditions on the outer boundary are satisfied approximately by applying the method of least-squares boundary collocation. A parametric study of finite-plate stress intensity factors, employing this mapping-collocation method, is presented. It shows the effects of varying material properties, orientation angle, and crack-length-to-plate-width and plate-height-to-plate-width ratios for rectangular orthotropic plates under constant tensile and shear loads.

Detection of License Plate using Sliding Window, Histogram of Oriented Gradient, and Support Vector Machines Method

NASA Astrophysics Data System (ADS)

Astawa, INGA; Gusti Ngurah Bagus Caturbawa, I.; Made Sajayasa, I.; Dwi Suta Atmaja, I. Made Ari

2018-01-01

The license plate recognition usually used as part of system such as parking system. License plate detection considered as the most important step in the license plate recognition system. We propose methods that can be used to detect the vehicle plate on mobile phone. In this paper, we used Sliding Window, Histogram of Oriented Gradient (HOG), and Support Vector Machines (SVM) method to license plate detection so it will increase the detection level even though the image is not in a good quality. The image proceed by Sliding Window method in order to find plate position. Feature extraction in every window movement had been done by HOG and SVM method. Good result had shown in this research, which is 96% of accuracy.

Hyperspectral imaging for detection of non-O157 Shiga-toxin producing Escherichia coli (STEC) serogroups on spread plates of mixed cultures

NASA Astrophysics Data System (ADS)

Yoon, Seung Chul; Windham, William R.; Ladely, Scott; Heitschmidt, Gerald W.; Lawrence, Kurt C.; Park, Bosoon; Narang, Neelam; Cray, William C.

2012-05-01

We investigated the feasibility of visible and near-infrared (VNIR) hyperspectral imaging for rapid presumptive-positive screening of six representative non-O157 Shiga-toxin producing Escherichia coli (STEC) serogroups (O26, O45, O103, O111, O121, and O145) on spread plates of mixed cultures. Although the traditional culture method is still the "gold standard" for presumptive-positive pathogen screening, it is time-consuming, labor-intensive, not effective in testing large amount of food samples, and cannot completely prevent unwanted background microflora from growing together with target microorganisms on agar media. A previous study was performed using the data obtained from pure cultures individually inoculated on spot and/or spread plates in order to develop multivariate classification models differentiating each colony of the six non-O157 STEC serogroups and to optimize the models in terms of parameters. This study dealt with the validation of the trained and optimized models with a test set of new independent samples obtained from colonies on spread plates of mixed cultures. A new validation protocol appropriate to a hyperspectral imaging study for mixed cultures was developed. One imaging experiment with colonies obtained from two serial dilutions was performed. A total of six agar plates were prepared, where O45, O111 and O121 serogroups were inoculated into all six plates and each of O45, O103 and O145 serogroups was added into the mixture of the three common bacterial cultures. The number of colonies grown after 24-h incubation was 331 and the number of pixels associated with the grown colonies was 16,379. The best model found from this validation study was based on pre-processing with standard normal variate and detrending (SNVD), first derivative, spectral smoothing, and k-nearest neighbor classification (kNN, k=3) of scores in the principal component subspace spanned by 6 principal components. The independent testing results showed 95% overall detection accuracy at pixel level and 97% at colony level. The developed model was proven to be still valid even for the independent samples although the size of a test set was small and only one experiment was performed. This study was an important first step in validating and updating multivariate classification models for rapid screening of ground beef samples contaminated by non-O157 STEC pathogens using hyperspectral imaging.

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.

In vivo evaluation of the effect of essential oil-containing oral strips on salivary bacteria using the checkerboard method.

PubMed

da Silva, Carina Maciel; Colombo, Andréa Vieira; do Souto, Renata Martins; Colombo, Ana Paula

2005-01-01

The aim of this study was to evaluate the antimicrobial effect of essential oil-containing oral strips on different species of the oral microbiota. Saliva samples were collected from 20 subjects with good oral health, diluted and plated onto blood agar medium. The subjects were asked to place the strip (Listerine PocketPaks) on the tongue allowing it to dissolve. After 30 minutes, new saliva samples were collected again and the plates with the samples were incubated under anaerobic conditions at 37 degrees C for seven days. Colony counts (CFU/mL) were determined for each sample. The colonies on the plates were washed with 1 mL of TE buffer, and the bacterial suspensions were processed for the identification of 24 species by DNA probes and the Checkerboard DNA-DNA hybridization method. Differences in total counts, prevalence, and levels of the species evaluated before and after placement of the strips were determined by Wilcoxon sign rank and Chi-square tests. A modest increase in the total bacterial number in saliva from 1.4 x 10(8) to 1.7 x 10(8) bacterial cells was observed 30 minutes after the strip placement, although this change was not significant (p = 0.632). Most of the species reduced in frequency and/or levels, including the pathogens A. actinomycetemcomitans, C. rectus, E. corrodens, Fusobacterium spp., P. intermedia, and S. noxia, as well as the beneficial species A. meyeri, A. georgia, A. gerencseriae, A. odontolyticus, and P. acnes after strip placement. In contrast, A. viscosus, P. melaninogenica, P. gingivalis, P. micros, Streptococcus spp., T. forsythensis, and V. parvula presented an increase in prevalence and/or levels. These changes were not statistically significant after adjusting for multiple comparisons (p > 0.0022). The use of the essential oil-containing oral strips resulted in a short-term small increase in the total number of salivary microorganisms. In addition, a not significant decrease of certain periodontopathogens, and an increase in species compatible with oral health were observed.

Models of convection-driven tectonic plates - A comparison of methods and results

NASA Technical Reports Server (NTRS)

King, Scott D.; Gable, Carl W.; Weinstein, Stuart A.

1992-01-01

Recent numerical studies of convection in the earth's mantle have included various features of plate tectonics. This paper describes three methods of modeling plates: through material properties, through force balance, and through a thin power-law sheet approximation. The results obtained are compared using each method on a series of simple calculations. From these results, scaling relations between the different parameterizations are developed. While each method produces different degrees of deformation within the surface plate, the surface heat flux and average plate velocity agree to within a few percent. The main results are not dependent upon the plate modeling method and herefore are representative of the physical system modeled.

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.

Evaluation of commercially prepared transport systems for nonlethal detection of Aeromonas salmonicida in salmonid fish

USGS Publications Warehouse

Cipriano, R.C.; Bullock, G.L.

2001-01-01

In vitro studies indicated that commercially prepared transport systems containing Amies, Stuart's, and Cary-Blair media worked equally well in sustaining the viability of the fish pathogen Aeromonas salmonicida, which causes furunculosis. The bacterium remained viable without significant increase or decrease in cell numbers for as long as 48 h of incubation at 18-20??C in Stuart's transport medium; consequently, obtaining mucus samples in such tubes were comparable to on-site detection of A. salmonicida by dilution plate counts on Coomassie Brilliant Blue agar. In three different assays of 100 samples of mucus from Atlantic salmon Salmo salar infected subclinically with A. salmonicida, dilution counts conducted on-site proved more reliable for detecting the pathogen than obtaining the samples in the transport system. In the on-site assays, dilution counts detected the pathogen in 34, 41, and 22 samples, whereas this was accomplished in only 15, 15, and 3 of the respective samples when the transport system was used. In an additional experiment, Arctic char Salvelinus alpinus sustaining a frank epizootic of furunculosis were sampled similarly. Here, too, dilution counts were more predictive of the prevalence of A. salmonicida and detected the pathogen in 46 mucus samples; in comparison, only 6 samples collected by using the transport system were positive. We also observed that the transport system supported the growth of the normal mucus bacterial flora. Particularly predominant among these were motile aeromonads and Pseudomonas fluorescens. In studies of mixed culture growth, two representatives of both of the latter genera of bacteria outgrew A. salmonicida - in some cases, to the total exclusion of the pathogen itself.

[Prokaryotic community of subglacial bottom sediments of Antarctic Lake Untersee: detection by cultural and direct microscopic techniques].

PubMed

Muliukin, A L; Demkina, E V; Manucharova, N A; Akimov, V N; Andersen, D; McKay, C; Gal'chenko, V F

2014-01-01

The heterotrophic mesophilic component was studied in microbial communities of the samples of frozen regolith collected from the glacier near Lake Untersee collected in 2011 during the joint Russian-American expedition to central Dronning Maud Land (Eastern Antarctica). Cultural techniques revealed high bacterial numbers in the samples. For enumeration of viable cells, the most probable numbers (MPN) method proved more efficient than plating on agar media. Fluorescent in situ hybridization with the relevant oligonucleotide probes revealed members of the groups Eubacteria (Actinobacteria, Firmicutes) and Archaea. Application of the methods of cell resuscitation, such as the use of diluted media and prevention of oxidative stress, did not result in a significant increase in the numbers of viable cells retrieved form subglacial sediment samples. Our previous investigations demonstrated the necessity for special procedures for efficient reactivation of the cells from microbial communities of preserved fossil soil and permafrost samples collected in the Arctic zone. The differences in response to the special resuscitation procedures may reflect the differences in the physiological and morphological state of bacterial cells in microbial communities subject to continuous or periodic low temperatures and dehydration.

[Screening and identification of a bacterium capable of converting agar to neoagaro oligosaccharides].

PubMed

Han, Junping; Huang, Yayan; Ye, Jing; Xiao, Meitian

2015-09-04

To screen and identify a bacterium capable of converting agar to neoagaro oligosaccharides. We took samples of porphyra haitanensis and nearby seawater, and then used the medium containing 1 per thousand agar to enrich the target bacteria. The target isolates were obtained by dilution-plate method, of which crude enzymes were further obtained by liquid culture. We adopted DNS method to determine the target bacteria which can convert agar to neoagaro oligosaccharides. The phylogenetics was identified by analyzing 16S rDNA sequence and combining the strain's morphological and bacterial colonial physiological biochemical characteristics. We isolated a gram-negative bacterial strain HJPHYXJ-1 capable of transforming agar to neoagaro oligosaccharides. Basic Local Alignment Search Tool (BLAST) search of HJPHYXJ-1's 16S rDNA sequence on GenBank suggested that the similarity between this strain and Vibrio natriegens reached 99% . In addition, the morphological and physiological biochemical characteristics of HJPHYXJ-1 also showed highly similarity to Vibrio natriegens. So we identified HJPHYXJ-1 as Vibrio natriegens. The results of HPLC suggested that the metabolite of enzymatic degradation was neoagaro oligosaccharides. HJPHYXJ-1 or the new isolate of Vibrio natriegens was capable of converting agar to neoagaro oligosaccharides.

Aseptic laboratory techniques: plating methods.

PubMed

Sanders, Erin R

2012-05-11

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: Perform plating procedures without contaminating media. Isolate single bacterial colonies by the streak-plating method. Use pour-plating and spread-plating methods to determine the concentration of bacteria. Perform soft agar overlays when working with phage. Transfer bacterial cells from one plate to another using the replica-plating procedure. Given an experimental task, select the appropriate plating method.

Coupling solid-phase extraction and enzyme-linked immunosorbent assay for ultratrace determination of herbicides in pristine water

USGS Publications Warehouse

Aga, D.S.; Thurman, E.M.

1993-01-01

Solid-phase extraction (SPE) and enzyme-linked immunosorbent assay (ELISA) were coupled for automated trace analysis of pristine water samples containing 2-chloro-4-ethylamino-6-isopropylamine-s-triazine (atrazine) and 2-chloro-2???,6???-diethyl-N-(methoxymethyl)acetanilide (alachlor). The isolation of the two herbicides on a C18-resin involved the selection of an elution solvent that both removes interfering substances and is compatible with ELISA. Ethyl acetate was selected as the elution solvent followed by a solvent exchange with methanol/water (20/80, % v/v). The SPE-ELISA method has a detection limit of 5.0 ng/L (5 ppt), >90% recovery, and a relative standard deviation of ??10%. The performance of a microtiter plate-based ELISA and a magnetic particle-based ELISA coupled to SPE was also evaluated. Although the sensitivity of the two ELISA methods was comparable, the precision using magnetic particles was improved considerably (??10% versus ??20%) because of the faster reaction kinetics provided by the magnetic particles. Finally, SPE-ELISA and isotope dilution gas chromatography/ mass spectrometry correlated well (correlation coefficient of 0.96) for lake-water samples. The SPE-ELISA method is simple and may have broader applications for the inexpensive automated analysis of other contaminants in water at trace levels.

Characterizing Novel Thermophilic Amylase Producing Bacteria From Taptapani Hot Spring, Odisha, India

PubMed Central

Sen, Sudip Kumar; Raut, Sangeeta; Satpathy, Soumya; Rout, Prangya Ranjan; Bandyopadhyay, Bidyut; Das Mohapatra, Pradeep Kumar

2014-01-01

Background: Amylases play a vital role in biotechnological studies and rank an important position in the world enzyme market (25% to 33%). Bioprocess method of amylase production is more effective than the other sources, since the technique is easy, cost effective, fast, and the enzymes of required properties can be procured. Objectives: The current study aimed to report the characteristics of novel amylase producing bacterial strains isolated from Taptapani hot spring, Odisha, India. Materials and Methods: Bacterial strains were isolated by dilution plating method from the water samples collected from Taptapani Hot Spring, Odisha and screened for amylase production through starch hydrolysis. The bacterial isolates were identified morphologically, biochemically, and finally by 16S rDNA profiling. Results: Based on the morphological, physiological, biochemical characteristics and the molecular characterization, the isolates SS1, SS2, and SS3 were identified as Bacillus barbaricus, Aeromonas veroni, and Stenotrophomonas maltophilia, respectively. The approximate molecular weight of enzymes from SS1, SS2, and SS3 strains were 19 kDa, 56 kDa and 49 kDa, respectively. Conclusions: The current report isolates, characterizes, and demonstrates the novel heat-adapted amylase-producing bacteria SS1, SS2 and SS3 from Taptapani hot spring, indicating its potentiality and stability under acidic conditions. PMID:25741425

Rifamycin action on RNA polymerase in antibiotic-tolerant Mycobacterium tuberculosis results in differentially detectable populations.

PubMed

Saito, Kohta; Warrier, Thulasi; Somersan-Karakaya, Selin; Kaminski, Lina; Mi, Jianjie; Jiang, Xiuju; Park, Suna; Shigyo, Kristi; Gold, Ben; Roberts, Julia; Weber, Elaina; Jacobs, William R; Nathan, Carl F

2017-06-13

Mycobacterium tuberculosis (Mtb) encounters stresses during the pathogenesis and treatment of tuberculosis (TB) that can suppress replication of the bacteria and render them phenotypically tolerant to most available drugs. Where studied, the majority of Mtb in the sputum of most untreated subjects with active TB have been found to be nonreplicating by the criterion that they do not grow as colony-forming units (cfus) when plated on agar. However, these cells are viable because they grow when diluted in liquid media. A method for generating such "differentially detectable" (DD) Mtb in vitro would aid studies of the biology and drug susceptibility of this population, but lack of independent confirmation of reported methods has contributed to skepticism about their existence. Here, we identified confounding artifacts that, when avoided, allowed development of a reliable method of producing cultures of ≥90% DD Mtb in starved cells. We then characterized several drugs according to whether they contribute to the generation of DD Mtb or kill them. Of the agents tested, rifamycins led to DD Mtb generation, an effect lacking in a rifampin-resistant strain with a mutation in rpoB , which encodes the canonical rifampin target, the β subunit of RNA polymerase. In contrast, thioridazine did not generate DD Mtb from starved cells but killed those generated by rifampin.

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

Evaluation of Two Standard and Two Chromogenic Selective Media for Optimal Growth and Enumeration of Isolates of 16 Unique Bacillus Species.

PubMed

Kabir, M Shahjahan; Hsieh, Ying-Hsin; Simpson, Steven; Kerdahi, Khalil; Sulaiman, Irshad M

2017-06-01

The genus Bacillus is a group of gram-positive endospore-forming bacteria that can cause food poisoning and diarrheal illness in humans. A wide range of food products have been linked to foodborne outbreaks associated with these opportunistic pathogens. The U.S. Food and Drug Administration recommends (in their Bacteriological Analytical Manual) the use of Bacara or mannitol egg yolk polymyxin (MYP) agar plates and the most-probable-number (MPN) method for enumeration and confirmation of Bacillus cereus and related species isolated from foods, sporadic cases, outbreaks, and routine environmental surveillance samples. We performed a comparative analysis of two chromogenic media (Bacara and Brilliance) and two traditional media (MYP and polymyxin egg yolk mannitol bromothymol blue agar [PEMBA]) for the isolation and enumeration of 16 Bacillus species under modified growth conditions that included pH, temperature, and dilution factor. A total of 50 environmental, food, and American Type Culture Collection reference isolates from 16 distinct Bacillus species were evaluated. A food adulteration experiment also was carried out by artificially adulterating two baby food matrices with two isolates each of B. cereus and Bacillus thuringiensis . Our results clearly indicated that chromogenic plating media (Bacara and Brilliance) are better than conventional standard media (MYP and PEMBA) for the detection and enumeration of B. cereus in foods and other official regulatory samples. The comparison of the two chromogenic media also indicated that Brilliance medium to be more efficient and selective for the isolation of Bacillus.

Digital PCR on a SlipChip.

PubMed

Shen, Feng; Du, Wenbin; Kreutz, Jason E; Fok, Alice; Ismagilov, Rustem F

2010-10-21

This paper describes a SlipChip to perform digital PCR in a very simple and inexpensive format. The fluidic path for introducing the sample combined with the PCR mixture was formed using elongated wells in the two plates of the SlipChip designed to overlap during sample loading. This fluidic path was broken up by simple slipping of the two plates that removed the overlap among wells and brought each well in contact with a reservoir preloaded with oil to generate 1280 reaction compartments (2.6 nL each) simultaneously. After thermal cycling, end-point fluorescence intensity was used to detect the presence of nucleic acid. Digital PCR on the SlipChip was tested quantitatively by using Staphylococcus aureus genomic DNA. As the concentration of the template DNA in the reaction mixture was diluted, the fraction of positive wells decreased as expected from the statistical analysis. No cross-contamination was observed during the experiments. At the extremes of the dynamic range of digital PCR the standard confidence interval determined using a normal approximation of the binomial distribution is not satisfactory. Therefore, statistical analysis based on the score method was used to establish these confidence intervals. The SlipChip provides a simple strategy to count nucleic acids by using PCR. It may find applications in research applications such as single cell analysis, prenatal diagnostics, and point-of-care diagnostics. SlipChip would become valuable for diagnostics, including applications in resource-limited areas after integration with isothermal nucleic acid amplification technologies and visual readout.

Rheological Properties of Commercially Available Hyaluronic Acid Products in the United States for the Treatment of Osteoarthritis Knee Pain

PubMed Central

Nicholls, Mathew; Manjoo, Ajay; Shaw, Peter; Niazi, Faizan; Rosen, Jeffrey

2018-01-01

Objective: The inconsistent results within the current literature regarding the efficacy of intra-articular-hyaluronic acid (IA-HA) for the treatment of knee osteoarthritis (OA) have been suggested to be due to intrinsic differences between individual HA products. The purpose of this investigation is to define the rheological differences between currently available HA products in the United States at the time of this study for the treatment of knee OA, which will help elaborate on the appropriateness of classifying HA products as a class opposed to as individual agents. Methods: The rheological parameters for Euflexxa, Orthovisc, Supartz, Monovisc, Synvisc, Synvisc-One, Gel-One, and Hyalgan were obtained with a TA AR 2000 EX Rheometer with a cone-plate geometry (40-mm plate diameter and a 2° cone angle) at room temperature. Results: The bulk rheological parameters of the different products suggest molecular structures traversing the range of dilute solution (Hyalgan, Supartz), semidilute solution (Euflexxa, Orthovisc), entangled solutions (Monovisc, Synvisc, Synvisc-One), and even gel-like (Gel-One) behavior. Conclusions: Due to the differences in rheological properties between IA-HA products, the universal assessment of these products as a class may not be appropriate. Instead, it may be more appropriate to assess each product individually. Future research should aim to link these differences in rheological properties to the differences in clinical efficacy seen across these IA-HA products. PMID:29326532

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.

Performance of dead Azolla filiculoides biomass in Biosorption of Au from wastewater.

PubMed

Umali, L J; Duncan, J R; Burgess, J E

2006-01-01

Dried milled biomass of Azolla filiculoides removed up to 98.2% of gold from wastewater from a gold plating factory containing 5 mg gold/l in solution in batch biosorption. The gold uptake capacity of the biomass was 98 mg/g. Whole dried biomass used in a continuous flow column removed up to 100% of gold from diluted wastewater. A similar column was linked to a sulphide precipitation process to provide a two-step system which was able to remove 98% of gold from undiluted wastewater containing 41 mg Au/l. The lifetime of the column was five days.

In vitro drug susceptibility pattern of Mycoplasma alligatoris isolated from symptomatic American alligators (Alligator mississippiensis).

PubMed

Helmick, Kelly E; Brown, Daniel R; Jacobson, Elliott R; Brown, Mary B

2002-06-01

A recently described mycoplasma, Mycoplasma alligatoris, was isolated from dead American alligators (Alligator mississippiensis) that had demonstrated clinical signs of lethargy, anorexia, bilateral ocular discharge, edema. paraparesis, and polyarthritis. The in vitro minimum inhibitory concentration for nine antibacterial agents was determined through serial dilution in broth and plate culture for M. alligatoris isolates. The inhibitory concentration obtained for doxycycline, enrofloxacin, sarafloxacin, oxytetracycline, tilmicosin, and tylosin (< 1 microg/ml) was lower than that of clindamycin (1-8 microg/ml), chloramphenicol (8-16 microg/ml), and erythromycin (32-138 microg/ml).

Thermal conductance measurements of bolted copper joints for SuperCDMS

DOE PAGES

Schmitt, R. L.; Tatkowski, G.; Ruschman, M.; ...

2015-04-28

Joint thermal conductance testing has been undertaken for bolted copper to copper connections from 60 mK to 26 K. This testing was performed to validate an initial design basis for the SuperCDMS experiment, where a dilution refrigerator will be coupled to a cryostat via multiple bolted connections. Copper used during testing was either gold plated or passivated with citric acid to prevent surface oxidation. Finally, the results we obtained are well fit by a power law regression of joint thermal conductance to temperature and match well with data collected during a literature review.

Automated food microbiology: potential for the hydrophobic grid-membrane filter.

PubMed Central

Sharpe, A N; Diotte, M P; Dudas, I; Michaud, G L

1978-01-01

Bacterial counts obtained on hydrophobic grid-membrane filters were comparable to conventional plate counts for Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus in homogenates from a range of foods. The wide numerical operating range of the hydrophobic grid-membrane filters allowed sequential diluting to be reduced or even eliminated, making them attractive as components in automated systems of analysis. Food debris could be rinsed completely from the unincubated hydrophobic grid-membrane filter surface without affecting the subsequent count, thus eliminating the possibility of counting food particles, a common source of error in electronic counting systems. PMID:100054

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.

Validation of high throughput screening of human sera for detection of anti-PA IgG by Enzyme-Linked Immunosorbent Assay (ELISA) as an emergency response to an anthrax incident

PubMed Central

Semenova, Vera A.; Steward-Clark, Evelene; Maniatis, Panagiotis; Epperson, Monica; Sabnis, Amit; Schiffer, Jarad

2017-01-01

To improve surge testing capability for a response to a release of Bacillus anthracis, the CDC anti-Protective Antigen (PA) IgG Enzyme-Linked Immunosorbent Assay (ELISA) was re-designed into a high throughput screening format. The following assay performance parameters were evaluated: goodness of fit (measured as the mean reference standard r2), accuracy (measured as percent error), precision (measured as coefficient of variance (CV)), lower limit of detection (LLOD), lower limit of quantification (LLOQ), dilutional linearity, diagnostic sensitivity (DSN) and diagnostic specificity (DSP). The paired sets of data for each sample were evaluated by Concordance Correlation Coefficient (CCC) analysis. The goodness of fit was 0.999; percent error between the expected and observed concentration for each sample ranged from −4.6% to 14.4%. The coefficient of variance ranged from 9.0% to 21.2%. The assay LLOQ was 2.6 μg/mL. The regression analysis results for dilutional linearity data were r2 = 0.952, slope = 1.02 and intercept = −0.03. CCC between assays was 0.974 for the median concentration of serum samples. The accuracy and precision components of CCC were 0.997 and 0.977, respectively. This high throughput screening assay is precise, accurate, sensitive and specific. Anti-PA IgG concentrations determined using two different assays proved high levels of agreement. The method will improve surge testing capability 18-fold from 4 to 72 sera per assay plate. PMID:27814939

Validation of high throughput screening of human sera for detection of anti-PA IgG by Enzyme-Linked Immunosorbent Assay (ELISA) as an emergency response to an anthrax incident.

PubMed

Semenova, Vera A; Steward-Clark, Evelene; Maniatis, Panagiotis; Epperson, Monica; Sabnis, Amit; Schiffer, Jarad

2017-01-01

To improve surge testing capability for a response to a release of Bacillus anthracis, the CDC anti-Protective Antigen (PA) IgG Enzyme-Linked Immunosorbent Assay (ELISA) was re-designed into a high throughput screening format. The following assay performance parameters were evaluated: goodness of fit (measured as the mean reference standard r 2 ), accuracy (measured as percent error), precision (measured as coefficient of variance (CV)), lower limit of detection (LLOD), lower limit of quantification (LLOQ), dilutional linearity, diagnostic sensitivity (DSN) and diagnostic specificity (DSP). The paired sets of data for each sample were evaluated by Concordance Correlation Coefficient (CCC) analysis. The goodness of fit was 0.999; percent error between the expected and observed concentration for each sample ranged from -4.6% to 14.4%. The coefficient of variance ranged from 9.0% to 21.2%. The assay LLOQ was 2.6 μg/mL. The regression analysis results for dilutional linearity data were r 2  = 0.952, slope = 1.02 and intercept = -0.03. CCC between assays was 0.974 for the median concentration of serum samples. The accuracy and precision components of CCC were 0.997 and 0.977, respectively. This high throughput screening assay is precise, accurate, sensitive and specific. Anti-PA IgG concentrations determined using two different assays proved high levels of agreement. The method will improve surge testing capability 18-fold from 4 to 72 sera per assay plate. Published by Elsevier Ltd.

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

Systems and Methods for the Electrodeposition of a Nickel-cobalt Alloy

NASA Technical Reports Server (NTRS)

Ogozalek, Nance Jo (Inventor); Wistrand, Richard E. (Inventor)

2013-01-01

Systems and methods for electrodepositing a nickel-cobalt alloy using a rotating cylinder electrode assembly with a plating surface and an electrical contact. The assembly is placed within a plating bath and rotated while running a plating cycle. Nickel-cobalt alloy deposition is selectively controlled by controlling current density distribution and/or cobalt content in the plating bath while running the plating cycle to deposit an alloy of a desired yield strength onto the plating surface in a single plating cycle. In various embodiments, the rotating cylinder may be used as an insitu monitoring method to assist in obtaining the properties desired.

Adherence of Aeromonas hydrophila strains to human enterocyte-like cells pre-infected with rotavirus.

PubMed

Bertuccio, M P; Picerno, I; Scoglio, M E

2012-09-01

The interest grown in these years about emerging pathogens in the onset of intestinal disease showed that the pathogenic mechanism is a multifactorial event. Our objective was to evaluate the role of co-infection with rotavirus in the expression of Aeromonas spp adhesiveness. The rate of co-infection involves contact of Caco-2 cells with the virus, followed by adsorption for 1 and 2 hours. Aliquots of bacterial suspensions were added to tissue-culture plates. After infection, cell monolayers were lysed; serially diluted lysates were plated to determine the number of bound bacteria by performing colony forming units (CFU) counts. Non-adhesive strains were not subject to variations resulting from co-infection, while those who had medium or high adhesiveness gave rise to an increase of the same. Infection with rotavirus promotes the Aeromonas ability to adhere to Caco-2 cells and this effect depends on the duration of infection and on the starting adhesiveness of bacteria strain.

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.

Aerial ultrasound source with a circular vibrating plate attached to a rigid circumferential wall

NASA Astrophysics Data System (ADS)

Kuratomi, Ryo; Asami, Takuya; Miura, Hikaru

2018-07-01

We fabricate a transverse vibrating plate attached to a rigid wall integrated at the circumference of a circular vibrating plate that allows a strong sound wave field to be formed in the area encoded by the vibrating plate and rigid wall by installing a wall such as a reflective plate on the rigid wall. The design method for the circular vibrating plate attached to a rigid circumferential wall is investigated. A method of forming a strong standing wave field in an enclosed area constructed with a vibrating plate, cylindrical reflective plate, and parallel reflective plate is developed.

[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

Cryopreservation on a cryo-plate of Arundina graminifolia protocorms, dehydrated with silica gel and drying beads.

PubMed

Cordova, L B; Thammasiri, K

2016-01-01

There are various methods for the cryopreservation of plant material, with each biological specimen potentially requiring protocol optimization to maximize success. The aim of this study is to compare droplet-vitrification, encapsulation-dehydration, and the cryo-plate method for cryopreservation of protocorms of the orchid Arundina graminifolia, using silica gel and drying beads as the desiccation materials. The cryo-plate method included preculture of protocorms, developed from seeds, placed on aluminium cryo-plates and embedded in alginate gel. Cryo-plates were surface dried using sterile filter paper, placed in Petri dishes containing 50 g silica gel or 30 g drying beads in a laminar air-flow cabinet. Specimens on cryo-plates were dehydrated to 25 % moisture content, placed into 2 mL cryotubes and plunged directly into liquid nitrogen for 1 d. For cryopreservation, the cryo-plate method, involving dehydration with 30 g drying beads gave the highest regrowth (77 %), followed by the encapsulation-dehydration method with 30 g drying beads (64 % regrowth) and the droplet-vitrification method, following exposure to PVS2 solution for 20 min (33 % regrowth). Regrowth of cryopreserved protocorms using the cryo-plate method was rapid with the highest survival and regrowth.

Examination of B. subtilis var. niger Spore Killing by Dry Heat Methods

NASA Technical Reports Server (NTRS)

Kempf, Michael J.; Kirschner, Larry E.

2004-01-01

Dry heat microbial reduction is the only NASA approved sterilization method to reduce the microbial bioburden on space-flight hardware prior to launch. Reduction of the microbial bioburden on spacecraft is necessary to meet planetary protection requirements specific for the mission. Microbial bioburden reduction also occurs if a spacecraft enters a planetary atmosphere (e.g., Mars) and is heated due to frictional forces. Temperatures reached during atmospheric entry events (>200 C) are sufficient to damage or destroy flight hardware and also kill microbial spores that reside on the in-bound spacecraft. The goal of this research is to determine the survival rates of bacterial spores when they are subjected to conditions similar to those the spacecraft would encounter (i.e., temperature, pressure, etc.). B. subtilis var. niger spore coupons were exposed to a range of temperatures from 125 C to 200 C in a vacuum oven (at <1 Torr). After the exposures, the spores were removed by sonication, dilutions were made, and the spores were plated using the pour plate method with tryptic soy agar. After 3 days incubation at 32 C, the number of colony-forming units was counted. Lethality rate constants and D-values were calculated at each temperature. The calculated D-values were: 27 minutes (at 125 C), 13 minutes (at 135 C), and <0.1 minutes (at 150 C). The 125 C and 135 C survivor curves appeared as concavedownward curves. The 150 C survivor curve appeared as a straight-line. Due to the prolonged ramp-up time to the exposure conditions, spore killing during the ramp-up resulted in insufficient data to draw curves for exposures at 160 C, 175 C, and 200 C. Exploratory experiments using novel techniques, with short ramp times, for performing high temperature exposures were also examined. Several of these techniques, such as vacuum furnaces, thermal spore exposure vessels, and laser heating of the coupons, will be discussed.

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.

Characterisation of structure-borne sound source using reception plate method.

PubMed

Putra, A; Saari, N F; Bakri, H; Ramlan, R; Dan, R M

2013-01-01

A laboratory-based experiment procedure of reception plate method for structure-borne sound source characterisation is reported in this paper. The method uses the assumption that the input power from the source installed on the plate is equal to the power dissipated by the plate. In this experiment, rectangular plates having high and low mobility relative to that of the source were used as the reception plates and a small electric fan motor was acting as the structure-borne source. The data representing the source characteristics, namely, the free velocity and the source mobility, were obtained and compared with those from direct measurement. Assumptions and constraints employing this method are discussed.

Consecutive Plate Acoustic Suppressor Apparatus and Methods

NASA Technical Reports Server (NTRS)

Doychak, Joseph (Inventor); Parrott, Tony L. (Inventor)

1993-01-01

An apparatus and method for suppressing acoustic noise utilizes consecutive plates, closely spaced to each other so as to exploit dissipation associated with sound propagation in narrow channels to optimize the acoustic resistance at a liner surface. The closely spaced plates can be utilized as high temperature structural materials for jet engines by constructing the plates from composite materials. Geometries of the plates, such as plate depth, shape, thickness, inter-plate spacing, arrangement, etc., can be selected to achieve bulk material-like behavior.

Nonlinear instability and convection in a vertically vibrated granular bed

NASA Astrophysics Data System (ADS)

Shukla, Priyanka; Ansari, I. H.; van der Meer, D.; Lohse, Detlef; Alam, Meheboob

2015-11-01

The nonlinear instability of the density-inverted granular Leidenfrost state and the resulting convective motion in strongly shaken granular matter are analysed via a weakly nonlinear analysis. Under a quasi-steady ansatz, the base state temperature decreases with increasing height away from from the vibrating plate, but the density profile consists of three distinct regions: (i) a collisional dilute layer at the bottom, (ii) a levitated dense layer at some intermediate height and (iii) a ballistic dilute layer at the top of the granular bed. For the nonlinear stability analysis, the nonlinearities up-to cubic order in perturbation amplitude are retained, leading to the Landau equation. The genesis of granular convection is shown to be tied to a supercritical pitchfork bifurcation from the Leidenfrost state. Near the bifurcation point the equilibrium amplitude is found to follow a square-root scaling law, Ae √{ ▵} , with the distance ▵ from bifurcation point. The strength of convection is maximal at some intermediate value of the shaking strength, with weaker convection both at weaker and stronger shaking. Our theory predicts a novel floating-convection state at very strong shaking.

Microbial structural diversity estimated by dilution-extinction of phenotypic traits and T-RFLP analysis along a land-use intensification gradient

NASA Technical Reports Server (NTRS)

Gomez, Elena del V.; Garland, Jay L.; Roberts, Michael S.

2004-01-01

The present work tested whether the relationship between functional traits and inoculum density reflected structural diversity in bacterial communities from a land-use intensification gradient applying a mathematical model. Terminal restriction fragment length polymorphism (T-RFLP) analysis was also performed to provide an independent assessment of species richness. Successive 10-fold dilutions of a soil suspension were inoculated onto Biolog GN(R) microplates. Soil bacterial density was determined by total cell and plate counts. The relationship between phenotypic traits and inoculum density fit the model, allowing the estimation of maximal phenotypic potential (Rmax) and inoculum density (KI) at which Rmax will be half-reduced. Though Rmax decreased with time elapsed since clearing of native vegetation, KI remained high in two of the disturbed sites. The genetic pool of bacterial community did not experience a significant reduction, but the active fraction responding in the Biolog assay was adversely affected, suggesting a reduction in the functional potential. c2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Aseptic Laboratory Techniques: Plating Methods

PubMed Central

Sanders, Erin R.

2012-01-01

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the streak-plating method. ● Use pour-plating and spread-plating methods to determine the concentration of bacteria. ● Perform soft agar overlays when working with phage. ● Transfer bacterial cells from one plate to another using the replica-plating procedure. ● Given an experimental task, select the appropriate plating method. PMID:22617405

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

Use of Coniothyrium minitans transformed with the hygromycin B resistance gene to study survival and infection of Sclerotinia sclerotiorum sclerotia in soil.

PubMed

Jones, E Eirian; Stewart, Alison; Whipps, John M

2003-03-01

A Coniothyrium minitans strain (T3) co-transformed with the genes for beta-glucuronidase (uidA) and hygromycin phosphotransferase (hph), the latter providing resistance to the antibiotic hygromycin B, was used to investigate the survival and infection of sclerotia of Sclerotinia sclerotiorum by C. minitans over time in four different soils. Infection of sclerotia was rapid in all cases, with the behaviour of transformant T3 and wild type parent A69 being similar. Differences were seen between the soils in the rate of infection of sclerotia by C. minitans and in their indigenous fungal populations. Amendment of agar with hygromycin B enabled the quantification of C. minitans in soil by dilution plating where there was a high background of other microorganisms. In Lincoln soil from New Zealand, which had a natural but low population of C. minitans, the hygromycin B resistance marker allowed the umambiguous discrimination of the applied transformed isolate from the indigenous hygromycin B sensitive one. In this soil, although the indigenous C. minitans population was detected from sclerotia, none were recovered on the dilution plates, indicating the increased sensitivity of C. minitans detection from soil using sclerotial baiting. C. minitans was a very efficient parasite, being able to infect a large proportion of sclerotia within a relatively short time from an initially low soil population. The addition of hygromycin B to agar also allowed the detection of C. minitans from decaying sclerotia by inhibiting secondary fungal colonisers. This is the first report to show that fungi colonising sclerotia already infected by C. minitans mask the detection of C. minitans from sclerotia rather than displacing the original parasite.

Protocols for dry DNA storage and shipment at room temperature.

PubMed

Ivanova, Natalia V; Kuzmina, Masha L

2013-09-01

The globalization of DNA barcoding will require core analytical facilities to develop cost-effective, efficient protocols for the shipment and archival storage of DNA extracts and PCR products. We evaluated three dry-state DNA stabilization systems: commercial Biomatrica(®) DNAstable(®) plates, home-made trehalose and polyvinyl alcohol (PVA) plates on 96-well panels of insect DNA stored at 56 °C and at room temperature. Controls included unprotected samples that were stored dry at room temperature and at 56 °C, and diluted samples held at 4 °C and at -20 °C. PCR and selective sequencing were performed over a 4-year interval to test the condition of DNA extracts. Biomatrica(®) provided better protection of DNA at 56 °C and at room temperature than trehalose and PVA, especially for diluted samples. PVA was the second best protectant after Biomatrica(®) at room temperature, whereas trehalose was the second best protectant at 56 °C. In spite of lower PCR success, the DNA stored at -20 °C yielded longer sequence reads and stronger signal, indicating that temperature is a crucial factor for DNA quality which has to be considered especially for long-term storage. Although it is premature to advocate a transition to DNA storage at room temperature, dry storage provides an additional layer of security for frozen samples, protecting them from degradation in the event of freezer failure. All three forms of DNA preservation enable shipment of dry DNA and PCR products between barcoding facilities. © 2013 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.

METHOD OF MAKING AND PLATE CONNECTION

DOEpatents

Patriarca, P.; Shubert, C.E.; Slaughter, G.M.

1963-02-26

This invention relates to a method of joining, by welding or brazing, a tube to a plate, particularly where the tube and the plate are of different thickness or have different thermal conductivities and are subject to high temperatures. In this method the tube is inserted in the core of a plate containing an annular groove in its back face concentric with the bore and in communication with the bore. One end of the tube is welded substantially flush with the front end of the plate. Brazing alloy is placed in the groove on the back face of the plate and heat is applied to the plate and tube to melt the brazing alloy to permit the alloy to flow into the bore and bond the tube to the plate. (AEC)

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.

Multiple internal seal ring micro-electro-mechanical system vacuum packaging method

NASA Technical Reports Server (NTRS)

Hayworth, Ken J. (Inventor); Yee, Karl Y. (Inventor); Shcheglov, Kirill V. (Inventor); Bae, Youngsam (Inventor); Wiberg, Dean V. (Inventor); Challoner, A. Dorian (Inventor); Peay, Chris S. (Inventor)

2008-01-01

A Multiple Internal Seal Ring (MISR) Micro-Electro-Mechanical System (MEMS) vacuum packaging method that hermetically seals MEMS devices using MISR. The method bonds a capping plate having metal seal rings to a base plate having metal seal rings by wafer bonding the capping plate wafer to the base plate wafer. Bulk electrodes may be used to provide conductive paths between the seal rings on the base plate and the capping plate. All seals are made using only metal-to-metal seal rings deposited on the polished surfaces of the base plate and capping plate wafers. However, multiple electrical feed-through metal traces are provided by fabricating via holes through the capping plate for electrical connection from the outside of the package through the via-holes to the inside of the package. Each metal seal ring serves the dual purposes of hermetic sealing and providing the electrical feed-through metal trace.

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.

Aluminum transfer method for plating plastics

NASA Technical Reports Server (NTRS)

Goodrich, W. D.; Stalmach, C. J., Jr.

1977-01-01

Electroless plating technique produces plate of uniform thickness. Hardness and abrasion resistance can be increased further by heat treatment. Method results in seamless coating over many materials, has low thermal conductivity, and is relatively inexpensive compared to conventional methods.

Application of the variational-asymptotical method to composite plates

NASA Technical Reports Server (NTRS)

Hodges, Dewey H.; Lee, Bok W.; Atilgan, Ali R.

1992-01-01

A method is developed for the 3D analysis of laminated plate deformation which is an extension of a variational-asymptotical method by Atilgan and Hodges (1991). Both methods are based on the treatment of plate deformation by splitting the 3D analysis into linear through-the-thickness analysis and 2D plate analysis. Whereas the first technique tackles transverse shear deformation in the second asymptotical approximation, the present method simplifies its treatment and restricts it to the first approximation. Both analytical techniques are applied to the linear cylindrical bending problem, and the strain and stress distributions are derived and compared with those of the exact solution. The present theory provides more accurate results than those of the classical laminated-plate theory for the transverse displacement of 2-, 3-, and 4-layer cross-ply laminated plates. The method can give reliable estimates of the in-plane strain and displacement distributions.

Chinese License Plates Recognition Method Based on A Robust and Efficient Feature Extraction and BPNN Algorithm

NASA Astrophysics Data System (ADS)

Zhang, Ming; Xie, Fei; Zhao, Jing; Sun, Rui; Zhang, Lei; Zhang, Yue

2018-04-01

The prosperity of license plate recognition technology has made great contribution to the development of Intelligent Transport System (ITS). In this paper, a robust and efficient license plate recognition method is proposed which is based on a combined feature extraction model and BPNN (Back Propagation Neural Network) algorithm. Firstly, the candidate region of the license plate detection and segmentation method is developed. Secondly, a new feature extraction model is designed considering three sets of features combination. Thirdly, the license plates classification and recognition method using the combined feature model and BPNN algorithm is presented. Finally, the experimental results indicate that the license plate segmentation and recognition both can be achieved effectively by the proposed algorithm. Compared with three traditional methods, the recognition accuracy of the proposed method has increased to 95.7% and the consuming time has decreased to 51.4ms.

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.

Inverse methods-based estimation of plate coupling in a plate motion model governed by mantle flow

NASA Astrophysics Data System (ADS)

Ratnaswamy, V.; Stadler, G.; Gurnis, M.

2013-12-01

Plate motion is primarily controlled by buoyancy (slab pull) which occurs at convergent plate margins where oceanic plates undergo deformation near the seismogenic zone. Yielding within subducting plates, lateral variations in viscosity, and the strength of seismic coupling between plate margins likely have an important control on plate motion. Here, we wish to infer the inter-plate coupling for different subduction zones, and develop a method for inferring it as a PDE-constrained optimization problem, where the cost functional is the misfit in plate velocities and is constrained by the nonlinear Stokes equation. The inverse models have well resolved slabs, plates, and plate margins in addition to a power law rheology with yielding in the upper mantle. Additionally, a Newton method is used to solve the nonlinear Stokes equation with viscosity bounds. We infer plate boundary strength using an inexact Gauss-Newton method with line search for backtracking. Each inverse model is applied to two simple 2-D scenarios (each with three subduction zones), one with back-arc spreading and one without. For each case we examine the sensitivity of the inversion to the amount of surface velocity used: 1) full surface velocity data and 2) surface velocity data simplified using a single scalar average (2-D equivalent to an Euler pole) for each plate. We can recover plate boundary strength in each case, even in the presence of highly nonlinear flow with extreme variations in viscosity. Additionally, we ascribe an uncertainty in each plate's velocity and perform an uncertainty quantification (UQ) through the Hessian of the misfit in plate velocities. We find that as plate boundaries become strongly coupled, the uncertainty in the inferred plate boundary strength decreases. For very weak, uncoupled subduction zones, the uncertainty of inferred plate margin strength increases since there is little sensitivity between plate margin strength and plate velocity. This result is significant because it implies we can infer which plate boundaries are more coupled (seismically) for a realistic dynamic model of plates and mantle flow.

Are electrostatic potentials between regions of different chemical composition measurable? The Gibbs-Guggenheim Principle reconsidered, extended and its consequences revisited.

PubMed

Pethica, Brian A

2007-12-21

As indicated by Gibbs and made explicit by Guggenheim, the electrical potential difference between two regions of different chemical composition cannot be measured. The Gibbs-Guggenheim Principle restricts the use of classical electrostatics in electrochemical theories as thermodynamically unsound with some few approximate exceptions, notably for dilute electrolyte solutions and concomitant low potentials where the linear limit for the exponential of the relevant Boltzmann distribution applies. The Principle invalidates the widespread use of forms of the Poisson-Boltzmann equation which do not include the non-electrostatic components of the chemical potentials of the ions. From a thermodynamic analysis of the parallel plate electrical condenser, employing only measurable electrical quantities and taking into account the chemical potentials of the components of the dielectric and their adsorption at the surfaces of the condenser plates, an experimental procedure to provide exceptions to the Principle has been proposed. This procedure is now reconsidered and rejected. No other related experimental procedures circumvent the Principle. Widely-used theoretical descriptions of electrolyte solutions, charged surfaces and colloid dispersions which neglect the Principle are briefly discussed. MD methods avoid the limitations of the Poisson-Bolzmann equation. Theoretical models which include the non-electrostatic components of the inter-ion and ion-surface interactions in solutions and colloid systems assume the additivity of dispersion and electrostatic forces. An experimental procedure to test this assumption is identified from the thermodynamics of condensers at microscopic plate separations. The available experimental data from Kelvin probe studies are preliminary, but tend against additivity. A corollary to the Gibbs-Guggenheim Principle is enunciated, and the Principle is restated that for any charged species, neither the difference in electrostatic potential nor the sum of the differences in the non-electrostatic components of the thermodynamic potential difference between regions of different chemical compositions can be measured.

Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI).

PubMed

Janetzki, Sylvia; Panageas, Katherine S; Ben-Porat, Leah; Boyer, Jean; Britten, Cedrik M; Clay, Timothy M; Kalos, Michael; Maecker, Holden T; Romero, Pedro; Yuan, Jianda; Kast, W Martin; Hoos, Axel

2008-03-01

The Cancer Vaccine Consortium of the Sabin Vaccine Institute (CVC/SVI) is conducting an ongoing large-scale immune monitoring harmonization program through its members and affiliated associations. This effort was brought to life as an external validation program by conducting an international Elispot proficiency panel with 36 laboratories in 2005, and was followed by a second panel with 29 participating laboratories in 2006 allowing for application of learnings from the first panel. Critical protocol choices, as well as standardization and validation practices among laboratories were assessed through detailed surveys. Although panel participants had to follow general guidelines in order to allow comparison of results, each laboratory was able to use its own protocols, materials and reagents. The second panel recorded an overall significantly improved performance, as measured by the ability to detect all predefined responses correctly. Protocol choices and laboratory practices, which can have a dramatic effect on the overall assay outcome, were identified and lead to the following recommendations: (A) Establish a laboratory SOP for Elispot testing procedures including (A1) a counting method for apoptotic cells for determining adequate cell dilution for plating, and (A2) overnight rest of cells prior to plating and incubation, (B) Use only pre-tested serum optimized for low background: high signal ratio, (C) Establish a laboratory SOP for plate reading including (C1) human auditing during the reading process and (C2) adequate adjustments for technical artifacts, and (D) Only allow trained personnel, which is certified per laboratory SOPs to conduct assays. Recommendations described under (A) were found to make a statistically significant difference in assay performance, while the remaining recommendations are based on practical experiences confirmed by the panel results, which could not be statistically tested. These results provide initial harmonization guidelines to optimize Elispot assay performance to the immunotherapy community. Further optimization is in process with ongoing panels.

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.

Assessment of a 96-Well Plate Assay of Quantitative Drug Susceptibility Testing for Mycobacterium Tuberculosis Complex in China.

PubMed

Xia, Hui; Zheng, Yang; Zhao, Bing; van den Hof, Susan; Cobelens, Frank; Zhao, YanLin

2017-01-01

To evaluate the performance of the Sensitire MYCOTB MIC Plate (MYCOTB) which could measure the twelve anti-tuberculosis drugs susceptibility on one 96-wells plate. A total of 140 MDR-TB strains and 60 non-MDR strains were sub-cultured and 193 strains were finally tested for drug resistance using MYCOTB and agar proportion method (APM) and another 7 strains failed of subculture. The drugs included ofloxacin (Ofx), moxifloxacin (Mfx), rifampin (RFP), amikacin (Am), rifabutin (Rfb), para-aminosalicylic acid (PAS), ethionamide (Eth), isoniazid (INH), kanamycin (Km), ethambutol (EMB), streptomycin (Sm), and cycloserine(Cs). The categorical agreement, conditional agreement, sensitivity and specificity of MYCOTB were assessed in comparison with APM. For strains with inconsistent results between MYCOTB and APM, the drug resistance related gene fragments were amplified and sequenced: gyrA for Ofx and Mfx; rpoB for RFP and Rfb; embB for EMB; rpsl for Sm; katG and the promoter region of inhA for INH, ethA and the promoter region of inhA for Eth. The sequence results were compared with results of MYCOTB and APM to analyze the consistency between sequence results and MYCOTB or APM. The categorical agreement between two methods for each drug ranged from 88.6% to 100%. It was the lowest for INH (88.6%). The sensitivity and specificity of MYCOTB ranged from 71.4% to 100% and 84.3% to 100%, respectively. The sensitivity was lowest for Cs(71.4%), EMB at 10μg/ml (80.0%) and INH at 10.0μg/ml (84.6%). The specificity was lowest for Rfb (84.3%). Overall discordance between the two phenotypic methods was observed for 96 strains, of which 63 (65.6%) were found susceptible with APM and resistant with MYCOTB and the remaining 33(34.4%) strains were resistant by APM and susceptible with MYCOTB. 34/52 (65.4%) sequenced APM susceptible and MYCOTB resistant(APM-S/MYCOTB-R) strains had mutations or insertions in the amplified regions. 20/30 (66.7%) sequenced APM resistant and MYCOTB susceptible strains had mutations in the sequenced genes. MICs of twenty-nine of these thirty isolates were equal to or within 1 doubling dilution of the critical concentration. MYCOTB had good performance for most of tested drugs and could be used as an alternative to the more labor demanding and longer turnaround time solid culture based DST method for detection of drug susceptibility in China.

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.

A novel tool for high-throughput screening of granulocyte-specific antibodies using the automated flow cytometric granulocyte immunofluorescence test (Flow-GIFT).

PubMed

Nguyen, Xuan Duc; Dengler, Thomas; Schulz-Linkholt, Monika; Klüter, Harald

2011-02-03

Transfusion-related acute lung injury (TRALI) is a severe complication related with blood transfusion. TRALI has usually been associated with antibodies against leukocytes. The flow cytometric granulocyte immunofluorescence test (Flow-GIFT) has been introduced for routine use when investigating patients and healthy blood donors. Here we describe a novel tool in the automation of the Flow-GIFT that enables a rapid screening of blood donations. We analyzed 440 sera from healthy female blood donors for the presence of granulocyte antibodies. As positive controls, 12 sera with known antibodies against anti-HNA-1a, -b, -2a; and -3a were additionally investigated. Whole-blood samples from HNA-typed donors were collected and the test cells isolated using cell sedimentation in a Ficoll density gradient. Subsequently, leukocytes were incubated with the respective serum and binding of antibodies was detected using FITC-conjugated antihuman antibody. 7-AAD was used to exclude dead cells. Pipetting steps were automated using the Biomek NXp Multichannel Automation Workstation. All samples were prepared in the 96-deep well plates and analyzed by flow cytometry. The standard granulocyte immunofluorescence test (GIFT) and granulocyte agglutination test (GAT) were also performed as reference methods. Sixteen sera were positive in the automated Flow-GIFT, while five of these sera were negative in the standard GIFT (anti-HNA 3a, n = 3; anti-HNA-1b, n = 1) and GAT (anti-HNA-2a, n = 1). The automated Flow-GIFT was able to detect all granulocyte antibodies, which could be only detected in GIFT in combination with GAT. In serial dilution tests, the automated Flow-GIFT detected the antibodies at higher dilutions than the reference methods GIFT and GAT. The Flow-GIFT proved to be feasible for automation. This novel high-throughput system allows an effective antigranulocyte antibody detection in a large donor population in order to prevent TRALI due to transfusion of blood products.

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.

A new license plate extraction framework based on fast mean shift

NASA Astrophysics Data System (ADS)

Pan, Luning; Li, Shuguang

2010-08-01

License plate extraction is considered to be the most crucial step of Automatic license plate recognition (ALPR) system. In this paper, a region-based license plate hybrid detection method is proposed to solve practical problems under complex background in which existing large quantity of disturbing information. In this method, coarse license plate location is carried out firstly to get the head part of a vehicle. Then a new Fast Mean Shift method based on random sampling of Kernel Density Estimate (KDE) is adopted to segment the color vehicle images, in order to get candidate license plate regions. The remarkable speed-up it brings makes Mean Shift segmentation more suitable for this application. Feature extraction and classification is used to accurately separate license plate from other candidate regions. At last, tilted license plate regulation is used for future recognition steps.

Biomechanical analysis using FEA and experiments of a standard plate method versus three cable methods for fixing acetabular fractures with simultaneous THA.

PubMed

Aziz, Mina S R; Dessouki, Omar; Samiezadeh, Saeid; Bougherara, Habiba; Schemitsch, Emil H; Zdero, Radovan

2017-08-01

Acetabular fractures potentially account for up to half of all pelvic fractures, while pelvic fractures potentially account for over one-tenth of all human bone fractures. This is the first biomechanical study to assess acetabular fracture fixation using plates versus cables in the presence of a total hip arthroplasty, as done for the elderly. In Phase 1, finite element (FE) models compared a standard plate method versus 3 cable methods for repairing an acetabular fracture (type: anterior column plus posterior hemi-transverse) subjected to a physiological-type compressive load of 2207N representing 3 x body weight for a 75kg person during walking. FE stress maps were compared to choose the most mechanically stable cable method, i.e. lowest peak bone stress. In Phase 2, mechanical tests were then done in artificial hemipelvises to compare the standard plate method versus the optimal cable method selected from Phase 1. FE analysis results showed peak bone stresses of 255MPa (Plate method), 205MPa (Mears cable method), 250MPa (Kang cable method), and 181MPa (Mouhsine cable method). Mechanical tests then showed that the Plate method versus the Mouhsine cable method selected from Phase 1 had higher stiffness (662versus 385N/mm, p=0.001), strength (3210versus 2060N, p=0.009), and failure energy (8.8versus 6.2J, p=0.002), whilst they were statistically equivalent for interfragmentary sliding (p≥0.179) and interfragmentary gapping (p≥0.08). The Plate method had superior mechanical properties, but the Mouhsine cable method may be a reasonable alternative if osteoporosis prevents good screw thread interdigitation during plating. Copyright © 2017 IPEM. Published by 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.

Effect of periodontal dressings on human gingiva fibroblasts in vitro

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

Eber, R.M.; Shuler, C.F.; Buchanan, W.

1989-08-01

In vitro cytotoxicity studies of periodontal dressings have not generally produced a result consistent with in vivo observations. These prior in vitro studies have not used human intraoral cell lines. We tested the effects of two eugenol containing and two non-eugenol periodontal dressings on cultured human gingival fibroblasts (HGF) (ATCC No. 1292). Replicate HGF cultures grown in microtiter plates were exposed to stock, 1:4 and 1:16 dilutions of extracts made from each of the four periodontal dressings. The HGF cultures were pulse labelled with tritiated thymidine (3HTdR) after 24, 48, and 72 hours. Incorporations of the labelled thymidine were measuredmore » using liquid scintillation counting and expressed as counts per minute. The results showed that undiluted extracts from all four periodontal dressings totally inhibited 3HTdR uptake (P less than 0.05). The 1:4 dilution of eugenol dressings inhibited 3HTdR uptake significantly more than non-eugenol dressings (P less than 0.05). Interestingly, at 72 hours the 1:16 dilution of the non-eugenol dressings caused significantly increased 3HTdR uptake which was not observed with the eugenol dressings. The present results suggest that the use of a human fibroblastic cell line for testing the effects of periodontal dressings may provide information about the relative biological effects of these dressings. Using this cell line, we have found that eugenol dressings inhibit fibroblast proliferation to a greater extent than non-eugenol dressings.« less

Electrochemical Assay of Gold-Plating Solutions

NASA Technical Reports Server (NTRS)

Chiodo, R.

1982-01-01

Gold content of plating solution is assayed by simple method that required only ordinary electrochemical laboratory equipment and materials. Technique involves electrodeposition of gold from solution onto electrode, the weight gain of which is measured. Suitable fast assay methods are economically and practically necessary in electronics and decorative-plating industries. If gold content in plating bath is too low, poor plating may result, with consequent economic loss to user.

[Determination of antiphospholipid antibodies in serum using ELISA].

PubMed

Fialová, L; Mikulíková, L; Malbohan, I; Průcha, M; Palecková, A; Cerný, V

1994-05-01

The authors compare two ELISA methods for the assessment of antiphospholipid antibodies, classes IgG and IgM, in serum: ELISA Pin Plate System ALPHA DIALAB Co. and the ELISA method developed in the Research Institute of Rheumatic Diseases. Both methods use cardiolipin as antigen. In the Pin Plate test the immunochemical reaction antigen/antibody does not take place at the surface of the pits of the microtitration plates but on the tip of the next plate. The results of examinations of antiphospholipid antibodies obtained by the tested methods are comparable, the Pin Plate test is quicker and more sensitive, but its price limits routine use.

Measurement of the residual stress distribution in a thick pre-stretched aluminum plate

NASA Astrophysics Data System (ADS)

Yuan, S. X.; Li, X. Q.; M, S.; Zhang, Y. C.; Gong, Y. D.

2008-12-01

Thick pre-stretched aluminum alloy plates are widely used in aircraft, while machining distortion caused by initial residual stress release in thick plates is a common and serious problem. To reduce the distortion, the residual stress distribution in thick plate must be measured. According to the characteristics of the thick pre-stretched aluminum alloy plate, based the elastic mechanical theory, this article deduces the modified layer-removal strain method adapting two different strain situations, which are caused by tensile and compressive stress. To validate this method, the residual stresses distribution along the thick direction of plate 2D70T351 is measured by this method, it is shown that the new method deduced in this paper is simple and accurate, and is very useful in engineering.

Application of coordinate transform on ball plate calibration

NASA Astrophysics Data System (ADS)

Wei, Hengzheng; Wang, Weinong; Ren, Guoying; Pei, Limei

2015-02-01

For the ball plate calibration method with coordinate measurement machine (CMM) equipped with laser interferometer, it is essential to adjust the ball plate parallel to the direction of laser beam. It is very time-consuming. To solve this problem, a method based on coordinate transformation between machine system and object system is presented. With the fixed points' coordinates of the ball plate measured in the object system and machine system, the transformation matrix between the coordinate systems is calculated. The laser interferometer measurement data error due to the placement of ball plate can be corrected with this transformation matrix. Experimental results indicate that this method is consistent with the handy adjustment method. It avoids the complexity of ball plate adjustment. It also can be applied to the ball beam calibration.

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.

Properties of large nearly perfect crystals at very low temperatures

NASA Technical Reports Server (NTRS)

Davis, W.; Krack, K. R.; Richard, J. P.; Weber, J.

1983-01-01

A liquid helium cryostat of a size and construction unavailable commercially, was built for use in measuring the Q of several materials at milli-Kelvin temperatures. The design and testing of the cryostat is described as well as the design of the experiment vacuum chamber and adaptor for the dilution refrigerator insert. Theory, design, and testing are also discussed for the magnetic coils built to levitate the materials so as to isolate them and increase the measured Q. A four point suspension with capacitor end plates as the transducer was used to obtain preliminary Q measurements of 6061 aluminum alloy and single crystal silicon. Results are tabulated.

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.

A comparison between orthogonal and parallel plating methods for distal humerus fractures: a prospective randomized trial.

PubMed

Lee, Sang Ki; Kim, Kap Jung; Park, Kyung Hoon; Choy, Won Sik

2014-10-01

With the continuing improvements in implants for distal humerus fractures, it is expected that newer types of plates, which are anatomically precontoured, thinner and less irritating to soft tissue, would have comparable outcomes when used in a clinical study. The purpose of this study was to compare the clinical and radiographic outcomes in patients with distal humerus fractures who were treated with orthogonal and parallel plating methods using precontoured distal humerus plates. Sixty-seven patients with a mean age of 55.4 years (range 22-90 years) were included in this prospective study. The subjects were randomly assigned to receive 1 of 2 treatments: orthogonal or parallel plating. The following results were assessed: operating time, time to fracture union, presence of a step or gap at the articular margin, varus-valgus angulation, functional recovery, and complications. No intergroup differences were observed based on radiological and clinical results between the groups. In our practice, no significant differences were found between the orthogonal and parallel plating methods in terms of clinical outcomes, mean operation time, union time, or complication rates. There were no cases of fracture nonunion in either group; heterotrophic ossification was found 3 patients in orthogonal plating group and 2 patients in parallel plating group. In our practice, no significant differences were found between the orthogonal and parallel plating methods in terms of clinical outcomes or complication rates. However, orthogonal plating method may be preferred in cases of coronal shear fractures, where posterior to anterior fixation may provide additional stability to the intraarticular fractures. Additionally, parallel plating method may be the preferred technique used for fractures that occur at the most distal end of the humerus.

Plating methods, a survey

NASA Technical Reports Server (NTRS)

Berkowitz, J. B.; Emerson, N. H.

1972-01-01

Results are presented of a comprehensive search of the literature available, much of which has been generated by the research centers of NASA and its contractors, on plating and coating methods and techniques. Methods covered included: (1) electroplating from aqueous solutions; (2) electroplating from nonaqueous solutions; (3) electroplating from fused-salt baths; (4) electroforming; (5) electroless plating, immersion plating, and mirroring; (6) electroplating from gaseous plasmas; and (7) anodized films and conversion coatings.

Mechanical design and analysis of focal plate for gravity deformation

NASA Astrophysics Data System (ADS)

Wang, Jianping; Chu, Jiaru; Hu, Hongzhuan; Li, Kexuan; Zhou, Zengxiang

2014-07-01

The surface accuracy of astronomical telescope focal plate is a key indicator to precision stellar observation. To conduct accurate deformation measurement for focal plate in different status, a 6-DOF hexapod platform was used for attitude adjustment. For the small adjustment range of a classic 6-DOF hexapod platform, an improved structural arrangement method was proposed in the paper to achieve ultimate adjustment of the focal plate in horizontal and vertical direction. To validate the feasibility of this method, an angle change model which used ball hinge was set up for the movement and base plate. Simulation results in MATLAB suggested that the ball hinge angle change of movement and base plate is within the range of the limiting angle in the process of the platform plate adjusting to ultimate attitude. The proposed method has some guiding significance for accurate surface measurement of focal plate.

Antibiotic Susceptibility of Neisseria gonorrhoeae Strains from Europe and Africa

PubMed Central

Piot, Peter; Van Dyck, Eddy; Colaert, Jan; Ursi, Jean-Paul; Bosmans, Eugène; Meheus, André

1979-01-01

The in vitro activities of 16 antimicrobial agents were tested by a plate dilution method against 268 unselected isolates of Neisseria gonorrhoeae from Belgium, Rwanda, Swaziland, and Zaire. Fifteen β-lactamase-producing strains isolated in Europe from various origins were also tested. There were significant regional variations in antimicrobial agent susceptibility, even among the African isolates, with the Rwandan and Zairean strains being most resistant. Benzylpenicillin and ampicillin were equally active in all but the β-lactamase-producing strains. Among the cephalosporins, cefotaxime was by far the most active, followed by cefuroxime, cefamandole, cefoxitin, and cefaclor, in that order. All strains were susceptible to spectinomycin, thiamphenicol, kanamycin, and rifampin, with the exception of one highly rifampin-resistant isolate and a moderately thiamphenicol-resistant strain. Twenty-six percent of the isolates were highly resistant to streptomycin. Six percent of the gonococci had a minimal inhibitory concentration for tetracycline greater than 2 μg/ml. Clavulanic acid inhibited the β-lactamase activity of the gonococci tested and improved markedly the activities of ampicillin and amoxicillin against β-lactamase-producing strains. PMID:111615

Antibacterial properties of amino acid functionalized silver nanoparticles decorated on graphene oxide sheets

NASA Astrophysics Data System (ADS)

Chandraker, Kumudini; Nagwanshi, Rekha; Jadhav, S. K.; Ghosh, Kallol K.; Satnami, Manmohan L.

2017-06-01

Graphene oxide (GO) sheets decorated with amino acid L-cysteine (L-cys) functionalized silver nanoparticles (GO-L-cys-Ag) was synthesized by AgNO3, trisodium citrate, and NaBH4. GO-L-cys-Ag nanocomposite was characterized by transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectra, ultraviolet-visible (UV-vis) absorption spectra, which demonstrated that a diameter of L-cys-AgNPs compactly deposited on GO. Antibacterial activity tests of GO-L-cys-Ag nanocomposite were carried out using Escherichia coli MTCC 1687 and Staphylococcus aureus MTCC 3160 as model strains of Gram-negative and Gram-positive bacteria, respectively. The effect of bactericide dosage on antibacterial activity of GO-L-cys-Ag nanocomposite was examined by plate count, well diffusion and broth dilution methods. Morphological observation of bacterial cells by scanning electron microscope (SEM) showed that GO-L-cys-Ag nanocomposite was more destructive to cell membrane of Escherichia coli than that of Staphylococcus aureus. The above technique establish that the bactericidal property of GO-L-cys-Ag nanocomposite with wide range of applications in biomedical science.

In vitro synergy testing of macrolide-quinolone combinations against 41 clinical isolates of Legionella.

PubMed Central

Martin, S J; Pendland, S L; Chen, C; Schreckenberger, P; Danziger, L H

1996-01-01

Combination antimicrobial therapy against Legionella species has not been well studied. Several quinolones have activity against Legionella strains, which prompted this in vitro search for a synergistic combination with the macrolides. By a checkerboard assay, erythromycin, clarithromycin, and azithromycin, each in combination with ciprofloxacin and levofloxacin, were tested for synergy against 46 isolates of Legionella. The agar dilution method was employed using buffered charcoal-yeast extract media. A final inoculum of 10(4) CFU per spot was prepared from 24-h growth of each isolate. Plates were incubated at 35 degrees C for 48 h. Synergy, partial synergy, additive effect, or indifference was observed for all combinations of antibiotics tested. There was no antagonism observed. Synergy occurred to a significantly greater extent for the clarithromycin-levofloxacin (P = 0.0001) and azithromycin-levofloxacin (P = 0.003) combinations versus erythromycin-levofloxacin. The azithromycin-ciprofloxacin combination demonstrated significantly greater synergy than did either erythromycin-ciprofloxacin (P = 0.003) or clarithromycin-ciprofloxacin (P = 0.001). The newer macrolides clarithromycin and azithromycin may be more active in combination with a fluoroquinolone than is erythromycin. PMID:8726012

Isolation Efficiency of Mouse Pancreatic Stem Cells Is Age Dependent.

PubMed

Kuise, Takashi; Noguchi, Hirofumi; Saitoh, Issei; Kataoka, Hitomi Usui; Watanabe, Masami; Noguchi, Yasufumi; Fujiwara, Toshiyoshi

2013-11-10

Mouse pancreatic stem cells have been isolated from mouse pancreata. This study evaluated the efficacy of isolating mouse pancreatic stem cells using mice of different ages. The pancreata of newborn mice, 8-week-old mice, and 24-week-old mice were harvested and digested by using collagenase. The "duct-like" cells in the digested pancreatic tissue were then inoculated into 96-well plates, cloned by limiting dilution, and cultured in DMEM with 20% FBS. Pancreatic stem cells were isolated from the pancreata of all newborn mice, while cells could only be isolated from 10% of the pancreata of 8-week-old mice and could not be isolated from the pancreata of any 24-week-old mice. These data suggest that young mice may have some pancreatic stem cells and that older mice may only have a few pancreatic stem cells. These data also indicate that it is extremely difficult to isolate pancreatic stem cells from older mice, suggesting that future research focus its efforts on finding methods of isolating pancreatic stem cells from adult mice.

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.

A computed microtomography method for understanding epiphyseal growth plate fusion

NASA Astrophysics Data System (ADS)

Staines, Katherine A.; Madi, Kamel; Javaheri, Behzad; Lee, Peter D.; Pitsillides, Andrew A.

2017-12-01

The epiphyseal growth plate is a developmental region responsible for linear bone growth, in which chondrocytes undertake a tightly regulated series of biological processes. Concomitant with the cessation of growth and sexual maturation, the human growth plate undergoes progressive narrowing, and ultimately disappears. Despite the crucial role of this growth plate fusion ‘bridging’ event, the precise mechanisms by which it is governed are complex and yet to be established. Progress is likely hindered by the current methods for growth plate visualisation; these are invasive and largely rely on histological procedures. Here we describe our non-invasive method utilising synchrotron x-ray computed microtomography for the examination of growth plate bridging, which ultimately leads to its closure coincident with termination of further longitudinal bone growth. We then apply this method to a dataset obtained from a benchtop microcomputed tomography scanner to highlight its potential for wide usage. Furthermore, we conduct finite element modelling at the micron-scale to reveal the effects of growth plate bridging on local tissue mechanics. Employment of these 3D analyses of growth plate bone bridging is likely to advance our understanding of the physiological mechanisms that control growth plate fusion.

New technical tip for anterior cervical plating : make hole first and choose the proper plate size later.

PubMed

Park, Jeong Yoon; Zhang, Ho Yeol; Oh, Min Chul

2011-04-01

It is well known that plate-to-disc distance (PDD) is closely related to adjacent-level ossification following anterior cervical plate placement. The study was undertaken to compare the outcomes of two different anterior cervical plating methods for degenerative cervical condition. Specifically, the new method involves making holes for plate screws first with an air drill and then choosing a plate size. The other method was standard, that is, decide on the plate size first, locate the plate on the anterior vertebral body, and then drilling the screw holes. Our null hypothesis was that the new technical tip may increase PDD as compared with the standard anterior cervical plating procedure. We retrospectively reviewed 49 patients who had a solid fusion after anterior cervical arthrodesis with a plate for the treatment of cervical disc degeneration. Twenty-three patients underwent the new anterior cervical plating technique (Group A) and 26 patients underwent the standard technique (Group B). PDD and ratios between PDD to anterior body heights (ABH) were measured using postoperative lateral radiographs. In addition, operating times and clinical results were reviewed in all cases. The mean durations of follow-up were 16.42±5.99 (Group A) and 19.83±6.71 (Group B) months, range 12 to 35 months. Of these parameters mentioned above, cephalad PDD (5.43 versus 3.46 mm, p=0.005) and cephalad PDD/ABH (0.36 versus 0.23, p=0.004) were significantly greater in the Group A, whereas operation time for two segment arthrodesis (141.9 versus 170.6 minutes, p=0.047) was significantly lower in the Group A. There were no significant difference between the two groups in caudal PDD (5.92 versus 5.06 mm), caudal PDD/ABH (0.37 versus 0.32) and clinical results. The new anterior cervical plating method represents an improvement over the standard method in terms of cephalad plate-to-disc distance and operating time.

Swelling characteristics of acrylic acid polyelectrolyte hydrogel in a dc electric field

NASA Astrophysics Data System (ADS)

Jabbari, Esmaiel; Tavakoli, Javad; Sarvestani, Alireza S.

2007-10-01

A novel application of environmentally sensitive polyelectrolytes is in the fabrication of BioMEMS devices as sensors and actuators. Poly(acrylic acid) (PAA) gels are anionic polyelectrolyte networks that exhibit volume expansion in aqueous physiological environments. When an electric field is applied to PAA polyelectrolyte gels, the fixed anionic polyelectrolyte charges and the requirement of electro-neutrality in the network generate an osmotic pressure, above that in the absence of the electric field, to expand the network. The objective of this research was to investigate the effect of an externally applied dc electric field on the volume expansion of the PAA polyelectrolyte gel in a simulated physiological solution of phosphate buffer saline (PBS). For swelling studies in the electric field, two platinum-coated plates, as electrodes, were wrapped in a polyethylene sheet to protect the plates from corrosion and placed vertically in a vessel filled with PBS. The plates were placed on a rail such that the distance between the two plates could be adjusted. The PAA gel was synthesized by free radical crosslinking of acrylic acid monomer with ethylene glycol dimethacrylate (EGDMA) crosslinker. Our results demonstrate that volume expansion depends on the intensity of the electric field, the PAA network density, network homogeneity, and the position of the gel in the field relative to positive/negative electrodes. Our model predictions for PAA volume expansion, based on the dilute electrolyte concentration in the gel network, is in excellent agreement with the experimental findings in the high-electric-field regime (250-300 Newton/Coulomb).

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.

A New Enzyme-Linked Immunosorbent Assay for a Total Anti-T Lymphocyte Globulin Determination: Development, Analytical Validation, and Clinical Applications.

PubMed

Montagna, Michela; La Nasa, Giorgio; Bernardo, Maria E; Piras, Eugenia; Avanzini, Maria A; Regazzi, Mario; Locatelli, Franco

2017-06-01

Anti-T lymphocyte globulin (ATLG) modulates the alloreactivity of T lymphocytes, reducing the risk of immunological posttransplant complications, in particular rejection and graft-versus-host disease, after allogeneic hematopoietic stem cell transplantation (HSCT). We developed and validated a new enzyme-linked immunosorbent assay (ELISA) method to measure serum levels of total ATLG and evaluate the pharmacokinetics (PK) of the drug in children with β-Thalassemia, receiving allogeneic HSCT. Diluted serum samples were incubated with Goat-anti-Rabbit IgG antibody coated on a microtiter plate and then, with Goat-anti-Human IgG labeled with horseradish peroxidase. After incubation and washings, substrate solution was added and absorbance was read at 492 nm. ATLG concentrations in samples were determined by interpolation from a standard curve (range: 200-0.095 ng/mL), prepared by diluting a known amount of ATLG in phosphate-buffered saline (PBS). Low, medium, and high-quality control concentrations were 1.56, 6.25, and 25 ng/mL, respectively. This method was developed and validated within the acceptance criteria in compliance with the Guidelines for a biological method validation: the sensitivity of the method was 0.095 ng/mL. We analyzed serum samples from 14 children with β-Thalassemia who received ATLG (Grafalon) at a dose of 10 mg/kg administered as intravenous (IV) infusion on days -5, -4, and -3 before HSCT (day 0). Blood sampling for PK evaluation was performed on days -5, -4, and -3 before and after drug infusion; and then from day -2 to +56. The median total ATLG levels pre-IVand post-IV were 0 and 118 mcg/mL on day -5; 85.9 and 199.2 mcg/mL on day -4; 153 and 270.9 mcg/mL on day -3, respectively. The median PK values of CL was 0.0029 (range: 0.0028-0.0057) L·kg·d, Vd was 0.088 (range: 0.025-0.448) L/kg and t1/2 was 20.2 (range: 5.8-50.2) days. These data suggest that given the marked interindividual variability of total ATLG disposition, the development of a validated ELISA method and the possibility to measure PK parameters in paediatric populations are essential steps to optimize drug therapeutic regimens.

Development and evaluation of the quick anaero-system-a new disposable anaerobic culture system.

PubMed

Yang, Nam Woong; Kim, Jin Man; Choi, Gwang Ju; Jang, Sook Jin

2010-04-01

We developed a new disposable anaerobic culture system, namely, the Quick anaero-system, for easy culturing of obligate anaerobes. Our system consists of 3 components: 1) new disposable anaerobic gas pack, 2) disposable culture-envelope and sealer, and 3) reusable stainless plate rack with mesh containing 10 g of palladium catalyst pellets. To evaluate the efficiency of our system, we used 12 anaerobic bacteria. We prepared 2 sets of ten-fold serial dilutions of the 12 anaerobes, and inoculated these samples on Luria-Bertani (LB) broth and LB blood agar plate (LB-BAP) (BD Diagnostic Systems, USA). Each set was incubated in the Quick anaero-system (DAS Tech, Korea) and BBL GasPak jar with BD GasPak EZ Anaerobe Container System (BD Diagnostic Systems) at 35-37 degrees C for 48 hr. The minimal inoculum size showing visible growth of 12 anaerobes when incubated in both the systems was compared. The minimal inoculum size showing visible growth for 2 out of the 12 anaerobes in the LB broth and 9 out of the 12 anaerobes on LB-BAP was lower for the Quick anaero-system than in the BD GasPak EZ Anaerobe Container System. The mean time (+/-SD) required to achieve absolute anaerobic conditions of the Quick anaero-system was 17 min and 56 sec (+/-3 min and 25 sec). The Quick anaero-system is a simple and effective method of culturing obligate anaerobes, and its performance is superior to that of the BD GasPak EZ Anaerobe Container System.

Microbes in Mascara: Hypothesis-Driven Research in a Nonmajor Biology Lab †

PubMed Central

Burleson, Kathryn M.; Martinez-Vaz, Betsy M.

2011-01-01

In this laboratory exercise, students were taught concepts of microbiology and scientific process through an everyday activity — cosmetic use. The students’ goals for the lab were to develop a hypothesis regarding microbial contamination in cosmetics, learn techniques to culture and differentiate microorganisms from cosmetics, and propose best practices in cosmetics use based on their findings. Prior to the lab, students took a pretest to assess their knowledge of scientific hypotheses, microbiology, and cosmetic safety. In the first week, students were introduced to microbiological concepts and methodologies, and cosmetic terminology and safety. Students completed a hypothesis-writing exercise before formulating and testing their own hypotheses regarding cosmetic contamination. Students provided a cosmetic of their own and, in consultation with their lab group, chose one product for testing. Samples were serially diluted and plated on a variety of selective media. In the second week, students analyzed their plates to determine the presence and diversity of microbes and if their hypotheses were supported. Students completed a worksheet of their results and were given a posttest to assess their knowledge. Average test scores improved from 5.2 (pretest) to 7.8 (posttest), with p-values < 0.0001. Seventy-nine percent (79%) of students correctly identified hypotheses that were not falsifiable or lacked variables, and 89% of students improved their scores on questions concerning safe cosmetic use. Ninety-one percent (91%) of students demonstrated increased knowledge of microbial concepts and methods. Based on our results, this lab is an easy, yet effective, way to enhance knowledge of scientific concepts for nonmajors, while maintaining relevance to everyday life. PMID:23653761

Lamb wave scattering by a surface-breaking crack in a plate

NASA Technical Reports Server (NTRS)

Datta, S. K.; Al-Nassar, Y.; Shah, A. H.

1991-01-01

An NDE method based on finite-element representation and modal expansion has been developed for solving the scattering of Lamb waves in an elastic plate waveguide. This method is very powerful for handling discontinuities of arbitrary shape, weldments of different orientations, canted cracks, etc. The advantage of the method is that it can be used to study the scattering of Lamb waves in anisotropic elastic plates and in multilayered plates as well.

Recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibody to turkey coronavirus.

PubMed

Abdelwahab, Mohamed; Loa, Chien Chang; Wu, Ching Ching; Lin, Tsang Long

2015-06-01

Nucleocapsid (N) protein gene of turkey coronavirus (TCoV) was expressed in a prokaryotic system and used to develop an enzyme-linked immunosorbent assay (ELISA) for detection of antibody to TCoV. Anti-TCoV hyperimmune turkey serum and normal turkey serum were used as positive or negative controls for optimization of the ELISA. Goat anti-turkey IgG (H+L) conjugated with horseradish peroxidase was used as detector antibody. Three hundred and twenty two turkey sera from the field were used to evaluate the performance of ELISA and determine the cut-off point of ELISA. The established ELISA was also examined with serum samples obtained from turkeys experimentally infected with TCoV. Those serum samples were collected at various time intervals from 1 to 63 days post-infection. The optimum conditions for differentiation between anti-TCoV hyperimmune serum and normal turkey serum were recombinant TCoV N protein concentration at 20 μg/ml, serum dilution at 1:800, and conjugate dilution at 1:10,000. Of the 322 sera from the field, 101 were positive for TCoV by immunofluorescent antibody assay (IFA). The sensitivity and specificity of the ELISA relative to IFA test were 86.0% and 96.8%, respectively, using the optimum cut-off point of 0.2 as determined by logistic regression method. Reactivity of anti-rotavirus, anti-reovirus, anti-adenovirus, or anti-enterovirus antibodies with the recombinant N protein coated on the ELISA plates was not detected. These results indicated that the established antibody-capture ELISA in conjunction with recombinant TCoV N protein as the coating protein can be utilized for detection of antibodies to TCoV in turkey flocks. Copyright © 2015 Elsevier B.V. All rights reserved.

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)

Estimating the number and size of phloem sieve plate pores using longitudinal views and geometric reconstruction.

PubMed

Bussières, Philippe

2014-05-12

Because it is difficult to obtain transverse views of the plant phloem sieve plate pores, which are short tubes, to estimate their number and diameters, a method based on longitudinal views is proposed. This method uses recent methods to estimate the number and the sizes of approximately circular objects from their images, given by slices perpendicular to the objects. Moreover, because such longitudinal views are obtained from slices that are rather close to the plate centres whereas the pore size may vary with the pore distance from the plate edge, a sieve plate reconstruction model was developed and incorporated in the method to consider this bias. The method was successfully tested with published longitudinal views of phloem of Soybean and an exceptional entire transverse view from the same tissue. The method was also validated with simulated slices in two sieve plates from Cucurbita and Phaseolus. This method will likely be useful to estimate and to model the hydraulic conductivity and the architecture of the plant phloem, and it could have applications for other materials with approximately cylindrical structures.

Rifamycin action on RNA polymerase in antibiotic-tolerant Mycobacterium tuberculosis results in differentially detectable populations

PubMed Central

Saito, Kohta; Warrier, Thulasi; Somersan-Karakaya, Selin; Kaminski, Lina; Mi, Jianjie; Jiang, Xiuju; Park, Suna; Shigyo, Kristi; Gold, Ben; Roberts, Julia; Weber, Elaina; Jacobs, William R.; Nathan, Carl F.

2017-01-01

Mycobacterium tuberculosis (Mtb) encounters stresses during the pathogenesis and treatment of tuberculosis (TB) that can suppress replication of the bacteria and render them phenotypically tolerant to most available drugs. Where studied, the majority of Mtb in the sputum of most untreated subjects with active TB have been found to be nonreplicating by the criterion that they do not grow as colony-forming units (cfus) when plated on agar. However, these cells are viable because they grow when diluted in liquid media. A method for generating such “differentially detectable” (DD) Mtb in vitro would aid studies of the biology and drug susceptibility of this population, but lack of independent confirmation of reported methods has contributed to skepticism about their existence. Here, we identified confounding artifacts that, when avoided, allowed development of a reliable method of producing cultures of ≥90% DD Mtb in starved cells. We then characterized several drugs according to whether they contribute to the generation of DD Mtb or kill them. Of the agents tested, rifamycins led to DD Mtb generation, an effect lacking in a rifampin-resistant strain with a mutation in rpoB, which encodes the canonical rifampin target, the β subunit of RNA polymerase. In contrast, thioridazine did not generate DD Mtb from starved cells but killed those generated by rifampin. PMID:28559332

Evaluation of dispersion methods for enumeration of microorganisms from peat and activated carbon biofilters treating volatile organic compounds.

PubMed

Khammar, Nadia; Malhautier, Luc; Degrange, Valérie; Lensi, Robert; Fanlo, Jean-Louis

2004-01-01

To enumerate microorganisms having colonized biofilters treating volatile organic compounds, it is necessary firstly to evaluate dispersion methods. Crushing, shaking and sonication were then tested for the removal of microflora from biofilters packing materials (peat and activated carbon). Continuous or discontinuous procedures, and addition of glass beads had no effect on the number of microorganisms removed from peat particles. The duration of treatment also had no effect for shaking and crushing, but the number of microorganisms after 60 min of treatment with ultrasound was significantly higher than that obtained after 0.5 min. The comparison between these methods showed that crushing was the most efficient for the removal of microorganisms from both peat and activated carbon. The comparison between three chemical dispersion agents showed that 1% Na-pyrophosphate was less efficient, compared with 200 mM phosphate buffer or 1% Na-hexametaphosphate. To optimize the cultivation of microorganisms, three different agar media were compared. Tryptic soy agar tenfold diluted (TSA 1/10) was the most suitable medium for the culture of microflora from a peat biofilter. For the activated carbon biofilter, there was no significant difference between Luria Bertoni, TSA 1/10, and plate count agar. The optimized extraction and enumeration protocols were used to perform a quantitative characterization of microbial populations in an operating laboratory activated carbon biofilter and in two parallel peat biofilters.

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

A New Method for Rapid Screening of End-Point PCR Products: Application to Single Genome Amplified HIV and SIV Envelope Amplicons

PubMed Central

Houzet, Laurent; Deleage, Claire; Satie, Anne-Pascale; Merlande, Laetitia; Mahe, Dominique; Dejucq-Rainsford, Nathalie

2015-01-01

PCR is the most widely applied technique for large scale screening of bacterial clones, mouse genotypes, virus genomes etc. A drawback of large PCR screening is that amplicon analysis is usually performed using gel electrophoresis, a step that is very labor intensive, tedious and chemical waste generating. Single genome amplification (SGA) is used to characterize the diversity and evolutionary dynamics of virus populations within infected hosts. SGA is based on the isolation of single template molecule using limiting dilution followed by nested PCR amplification and requires the analysis of hundreds of reactions per sample, making large scale SGA studies very challenging. Here we present a novel approach entitled Long Amplicon Melt Profiling (LAMP) based on the analysis of the melting profile of the PCR reactions using SYBR Green and/or EvaGreen fluorescent dyes. The LAMP method represents an attractive alternative to gel electrophoresis and enables the quick discrimination of positive reactions. We validate LAMP for SIV and HIV env-SGA, in 96- and 384-well plate formats. Because the melt profiling allows the screening of several thousands of PCR reactions in a cost-effective, rapid and robust way, we believe it will greatly facilitate any large scale PCR screening. PMID:26053379

[Study on the Chinese herbal formula for treatment of vaginitis and the antimicrobial activity in murine models].

PubMed

Fu, Ting-ting; Wu, Jian-yuan; Wang, Li; Ma, Yao; Wang, Ying; Liu, Ying; Ding, Hong

2006-09-01

To study on the various proportions of Radix Sophorae Flavescentis, Cortex Phellodendri, Fructus Cnidii and pericarp of Zanthoxylum bungeanum Maxim in the formulas, whose antimicrobial effects on E. coli, S. aureus, P. aeruginosa and C. albicans under different pH values were compared in vitro. According to Chinese ancient proved recipe, the K-B method and plate diluting method were adopted to measure antimicrobial activity, and orthogonal design to ascertain the herbal formula in vitro. Finally, murine models were established to test the antimicrobial activity in vivo through vaginal membrane irritancy experiment, negative rate of pathogeny and pathological grade of vaginal membrane. The results suggested that formulas with different proportions of the herbs had diverse antimicrobial activities, and the effect was shown to be most obvious when one milliliter drug contains 100 microl Fructus Cnidii-pericarp of Zanthoxylum bungeanum (2:1) co-extracted volatile oil and 50 microl Radix Sophorae Flavescentis and Cortex Phenodendri ethanol extraction respectively under pH6. The antimicrobial effect of the formula, which hardly had any membrane irritancy, was better than Jie Eryin in vitro and vivo. The fromula has few components and better effect, and adaptation to the pH value of vaginitis. It is a promising alternative for gynecological diseases.

Pyroelectricity Assisted Infrared-Laser Desorption Ionization (PAI-LDI) for Atmospheric Pressure Mass Spectrometry

NASA Astrophysics Data System (ADS)

Li, Yanyan; Ma, Xiaoxiao; Wei, Zhenwei; Gong, Xiaoyun; Yang, Chengdui; Zhang, Sichun; Zhang, Xinrong

2015-08-01

A new atmospheric pressure ionization method termed pyroelectricity-assisted infrared laser desorption ionization (PAI-LDI) was developed in this study. The pyroelectric material served as both sample target plate and enhancing ionization substrate, and an IR laser with wavelength of 1064 nm was employed to realize direct desorption and ionization of the analytes. The mass spectra of various compounds obtained on pyroelectric material were compared with those of other substrates. For the five standard substances tested in this work, LiNbO3 substrate produced the highest ion yield and the signal intensity was about 10 times higher than that when copper was used as substrate. For 1-adamantylamine, as low as 20 pg (132.2 fmol) was successfully detected. The active ingredient in (Compound Paracetamol and 1-Adamantylamine Hydrochloride Capsules), 1-adamantylamine, can be sensitively detected at an amount as low as 150 pg, when the medicine stock solution was diluted with urine. Monosaccharide and oligosaccharides in Allium Cepa L. juice was also successfully identified with PAI-LDI. The method did not require matrix-assisted external high voltage or other extra facility-assisted set-ups for desorption/ionization. This study suggested exciting application prospect of pyroelectric materials in matrix- and electricity-free atmospheric pressure mass spectrometry research.

Clinical comparison of two different plating methods in minimally invasive plate osteosynthesis for clavicular midshaft fractures: A randomized controlled trial.

PubMed

Sohn, Hoon-Sang; Shon, Min Soo; Lee, Kyung-Hag; Song, Si-Jung

2015-11-01

The aim of this study was to compare the clinical and radiographic outcomes between two different plating methods (superior vs. anteroinferior) in minimally invasive plate osteosynthesis (MIPO) for acute displaced clavicular shaft fractures. A prospective, randomized controlled trial was performed in a single centre. Nineteen patients were treated with superior plating and 18 with anteroinferior plating using the MIPO technique. A 3.5-mm locking reconstruction plate was bent preoperatively and applied to either the anteroinferior or superior aspect of the clavicle through two separate incisions. The operating time, time to union, the proportional length difference, complications, and functional outcome of the shoulder joint were evaluated using the Constant score and the University of California Los Angeles (UCLA) score. There was no statistically significant difference in the Constant score and UCLA score. The mean time to union was 16.8 weeks for superior plating and 17.1 weeks for anteroinferior plating (p=0.866). The average operation time was 77.2min in superior plating and 79.4min in anteroinferior plating (p=0.491). One patient in the superior plating group showed plate failure. Despite no significant difference, one patient had nonunion in the superior plating group (p>0.999). From a clinical perspective, although MIPO with anteroinferior plating provides better outcomes especially in complications without statistically significant difference, both plating methods provided satisfactory clinical and radiographic outcomes. Level I, a single-centre, prospective, randomized controlled trial. Copyright © 2015 Elsevier Ltd. All rights reserved.

Method of ultrasonic measurement of texture

DOEpatents

Thompson, R. Bruce; Smith, John F.; Lee, Seung S.; Li, Yan

1993-10-12

A method for measuring texture of metal plates or sheets using non-destructive ultrasonic investigation includes measuring the velocity of ultrasonic energy waves in lower order plate modes in one or more directions, and measuring phase velocity dispersion of higher order modes of the plate or sheet if needed. Texture or preferred grain orientation can be derived from these measurements with improves reliability and accuracy. The method can be utilized in production on moving metal plate or sheet.

Method of ultrasonic measurement of texture

DOEpatents

Thompson, R.B.; Smith, J.F.; Lee, S.S.; Taejon Ch'ungmam; Yan Li.

1993-10-12

A method for measuring texture of metal plates or sheets using non-destructive ultrasonic investigation includes measuring the velocity of ultrasonic energy waves in lower order plate modes in one or more directions, and measuring phase velocity dispersion of higher order modes of the plate or sheet if needed. Texture or preferred grain orientation can be derived from these measurements with improves reliability and accuracy. The method can be utilized in production on moving metal plate or sheet. 9 figures.

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.

Optimum stacking sequence design of laminated composite circular plates with curvilinear fibres by a layer-wise optimization method

NASA Astrophysics Data System (ADS)

Guenanou, A.; Houmat, A.

2018-05-01

The optimum stacking sequence design for the maximum fundamental frequency of symmetrically laminated composite circular plates with curvilinear fibres is investigated for the first time using a layer-wise optimization method. The design variables are two fibre orientation angles per layer. The fibre paths are constructed using the method of shifted paths. The first-order shear deformation plate theory and a curved square p-element are used to calculate the objective function. The blending function method is used to model accurately the geometry of the circular plate. The equations of motion are derived using Lagrange's method. The numerical results are validated by means of a convergence test and comparison with published values for symmetrically laminated composite circular plates with rectilinear fibres. The material parameters, boundary conditions, number of layers and thickness are shown to influence the optimum solutions to different extents. The results should serve as a benchmark for optimum stacking sequences of symmetrically laminated composite circular plates with curvilinear fibres.

Understanding the solidification and microstructure evolution during CSC-MIG welding of Fe–Cr–B-based alloy

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

Sorour, A.A., E-mail: ahmad.sorour@mail.mcgill.ca; Chromik, R.R., E-mail: richard.chromik@mcgill.ca; Gauvin, R., E-mail: raynald.gauvin@mcgill.ca

2013-12-15

The present is a study of the solidification and microstructure of Fe–28.2%Cr–3.8%B–1.5%Si–1.5%Mn (wt.%) alloy deposited onto a 1020 plain carbon steel substrate using the controlled short-circuit metal inert gas welding process. The as-solidified alloy was a metal matrix composite with a hypereutectic microstructure. Thermodynamic calculation based on the Scheil–Gulliver model showed that a primary (Cr,Fe){sub 2}B phase formed first during solidification, followed by an eutectic formation of the (Cr,Fe){sub 2}B phase and a body-centered cubic Fe-based solid solution matrix, which contained Cr, Mn and Si. Microstructure analysis confirmed the formation of these phases and showed that the shape of themore » (Cr,Fe){sub 2}B phase was irregular plate. As the welding heat input increased, the weld dilution increased and thus the volume fraction of the (Cr,Fe){sub 2}B plates decreased while other microstructural characteristics were similar. - Highlights: • We deposit Fe–Cr–B-based alloy onto plain carbon steel using the CSC-MIG process. • We model the solidification behavior using thermodynamic calculation. • As deposited alloy consists of (Cr,Fe){sub 2}B plates embedded in Fe-based matrix. • We study the effect of the welding heat input on the microstructure.« less

Experimental analysis of a Flat Plate Pulsating Heat Pipe with Self-ReWetting Fluids during a parabolic flight campaign

NASA Astrophysics Data System (ADS)

Cecere, Anselmo; De Cristofaro, Davide; Savino, Raffaele; Ayel, Vincent; Sole-Agostinelli, Thibaud; Marengo, Marco; Romestant, Cyril; Bertin, Yves

2018-06-01

A Flat Plate Pulsating Heat Pipe (FPPHP) filled with an ordinary liquid (water) and a self-rewetting mixture (dilutes aqueous solutions of long-chain alcohols with unusual surface tension behavior) is investigated under variable gravity conditions on board a 'Zero-g' plane during the 65th Parabolic Flight Campaign of the European Space Agency. The FPPHP thermal performance in terms of evaporator and condenser temperatures, start-up levels and flow regimes is characterized for the two working fluids and a power input ranging from 0 to 200 W (up to 17 W/cm2 at the heater/evaporator wall interface). The experimental set-up also includes a transparent plate enabling the visualization of the oscillating flow patterns during the experiments. For a low power input (4 W/cm2), the pulsating heat pipe filled with pure water is not able to work under low-g conditions, because the evaporator immediately exhibits dry-out conditions and the fluid oscillations stops, preventing heat transfer between the hot and cold side and resulting in a global increase of the temperatures. On the other hand, the FPPHP filled with the self-rewetting fluid runs also during the microgravity phase. The liquid rewets several times the evaporator zone triggering the oscillatory regime. The self-rewetting fluid helps both the start-up and the thermal performance of the FPPHP in microgravity conditions.

Changes in the mycovirus (LeV) titer and viral effect on the vegetative growth of the edible mushroom Lentinula edodes.

PubMed

Kim, Jung-Mi; Song, Ha-Yeon; Choi, Hyo-Jin; Yun, Suk-Hyun; So, Kum-Kang; Ko, Han-Kyu; Kim, Dae-Hyuk

2015-02-02

This study attempted to cure the edible mushroom Lentinula edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. These results indicate that LeV infection has a deleterious effect on mycelial growth. Copyright © 2014 Elsevier B.V. All rights reserved.

In Vitro Cultivation of 'Unculturable' Oral Bacteria, Facilitated by Community Culture and Media Supplementation with Siderophores.

PubMed

Vartoukian, Sonia R; Adamowska, Aleksandra; Lawlor, Megan; Moazzez, Rebecca; Dewhirst, Floyd E; Wade, William G

2016-01-01

Over a third of oral bacteria are as-yet-uncultivated in-vitro. Siderophores have been previously shown to enable in-vitro growth of previously uncultivated bacteria. The objective of this study was to cultivate novel oral bacteria in siderophore-supplemented culture media. Various compounds with siderophore activity, including pyoverdines-Fe-complex, desferricoprogen and salicylic acid, were found to stimulate the growth of difficult-to-culture strains Prevotella sp. HOT-376 and Fretibacterium fastidiosum. Furthermore, pyrosequencing analysis demonstrated increased proportions of the as-yet-uncultivated phylotypes Dialister sp. HOT-119 and Megasphaera sp. HOT-123 on mixed culture plates supplemented with siderophores. Therefore a culture model was developed, which incorporated 15 μg siderophore (pyoverdines-Fe-complex or desferricoprogen) or 150 μl neat subgingival-plaque suspension into a central well on agar plates that were inoculated with heavily-diluted subgingival-plaque samples from subjects with periodontitis. Colonies showing satellitism were passaged onto fresh plates in co-culture with selected helper strains. Five novel strains, representatives of three previously-uncultivated taxa (Anaerolineae bacterium HOT-439, the first oral taxon from the Chloroflexi phylum to have been cultivated; Bacteroidetes bacterium HOT-365; and Peptostreptococcaceae bacterium HOT-091) were successfully isolated. All novel isolates required helper strains for growth, implying dependence on a biofilm lifestyle. Their characterisation will further our understanding of the human oral microbiome.

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.

[Screening and antibacterial function of Bacillus amyloliquefaciens X030].

PubMed

He, Hao; Zhu, Yingling; Chi, Liqing; Zhao, Zizhao; Wang, Ting; Zuo, Mingxing; Zhang, Tong; Zhou, Fengjuan; Xia, Liqiu; Ding, Xuezhi

2015-09-04

We isolated 339 bacillus strains from 72 soil samples all over the country, then purified their antimicrobial compounds and studied the antibacterial activity, to enrich bacillus resources and explore their second metabolites. A bacillus strain with strong antibacterial activity was selected by dilution plate and water bath heating from a soil sample from a peanut plantation in Henan Province; this strain was identified according to morphological observation, physiological and biochemical characteristics, and consequences of 16S rRNA homologous analysis. Antibacterial compound from the identified strain, Bacillus amyloliquefaciens X030, was separated and purified by acetone precipitation, Sephadex chromatography, C18 reverse phase column chromatography. Its molecular weight was analyzed by LC-MS/MS. The antibacterial activity was characterized by disc diffusion and plate two-way cultivation. Bacillus amyloliquefaciens was isolated that not only has antibacterial activity against Staphylococcus aureus, Candida albican and Saccharomycetes; but also against Pyriculariaoryzae, Chili pointed cell anthrax, Gloeosporium eriobotryae speg and Phytophthora parasitica. The compound was confirmed as polypeptide. Bacillus amyloliquefaciens X030 can produce a polypeptide that inhibits pathogenic bacteria and plant pathogenic fungi.

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.

The constraint method: A new finite element technique. [applied to static and dynamic loads on plates

NASA Technical Reports Server (NTRS)

Tsai, C.; Szabo, B. A.

1973-01-01

An approch to the finite element method which utilizes families of conforming finite elements based on complete polynomials is presented. Finite element approximations based on this method converge with respect to progressively reduced element sizes as well as with respect to progressively increasing orders of approximation. Numerical results of static and dynamic applications of plates are presented to demonstrate the efficiency of the method. Comparisons are made with plate elements in NASTRAN and the high-precision plate element developed by Cowper and his co-workers. Some considerations are given to implementation of the constraint method into general purpose computer programs such as NASTRAN.

METHOD OF APPLYING COPPER COATINGS TO URANIUM

DOEpatents

Gray, A.G.

1959-07-14

A method is presented for protecting metallic uranium, which comprises anodic etching of the uranium in an aqueous phosphoric acid solution containing chloride ions, cleaning the etched uranium in aqueous nitric acid solution, promptly electro-plating the cleaned uranium in a copper electro-plating bath, and then electro-plating thereupon lead, tin, zinc, cadmium, chromium or nickel from an aqueous electro-plating bath.

A THUMBNAIL HISTORY OF HETEROTROPHIC PLATE COUNT (HPC) METHODOLOGY IN THE UNITED STATES

EPA Science Inventory

Over the past 100 years, the method of determining the number of bacteria in water, foods or other materials has been termed variously as: bacterial plate count, total plate count, total viable plate count, aerobic plate count, standard plate cound and more recently, heterotrophi...

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

Finding Platinum-Coating Gaps On Titanium Anodes

NASA Technical Reports Server (NTRS)

Bodemeijer, Ronnald; Flowers, Cecil E.

1990-01-01

Simple procedure makes gaps visible to eye. New gap-detection method consists of plating thin layer of non-silver-colored metal like copper or gold on anode. Contrast in color between plated metal and bare anode material makes gaps stand out. If anode passes inspection, copper or gold plate removable by reversal of test-plating current. Remains to be determined whether test plating and removal damages anode. New method simpler and more economical than previous attempts to identify gaps in platinum.

Fuel cell design and assembly

DOEpatents

Myerhoff, Alfred

1984-01-01

The present invention is directed to a novel bipolar cooling plate, fuel cell design and method of assembly of fuel cells. The bipolar cooling plate used in the fuel cell design and method of assembly has discrete opposite edge and means carried by the plate defining a plurality of channels extending along the surface of the plate toward the opposite edges. At least one edge of the channels terminates short of the edge of the plate defining a recess for receiving a fastener.

The vibroacoustic response and sound absorption performance of multilayer, microperforated rib-stiffened plates

NASA Astrophysics Data System (ADS)

Zhou, Haian; Wang, Xiaoming; Wu, Huayong; Meng, Jianbing

2017-10-01

The vibroacoustic response and sound absorption performance of a structure composed of multilayer plates and one rigid back wall are theoretically analyzed. In this structure, all plates are two-dimensional, microperforated, and periodically rib-stiffened. To investigate such a structural system, semianalytical models of one-layer and multilayer plate structures considering the vibration effects are first developed. Then approaches of the space harmonic method and Fourier transforms are applied to a one-layer plate, and finally the cascade connection method is utilized for a multilayer plate structure. Based on fundamental acoustic formulas, the vibroacoustic responses of microperforated stiffened plates are expressed as functions of a series of harmonic amplitudes of plate displacement, which are then solved by employing the numerical truncation method. Applying the inverse Fourier transform, wave propagation, and linear addition properties, the equations of the sound pressures and absorption coefficients for the one-layer and multilayer stiffened plates in physical space are finally derived. Using numerical examples, the effects of the most important physical parameters—for example, the perforation ratio of the plate, sound incident angles, and periodical rib spacing—on sound absorption performance are examined. Numerical results indicate that the sound absorption performance of the studied structure is effectively enhanced by the flexural vibration of the plate in water. Finally, the proposed approaches are validated by comparing the results of stiffened plates of the present work with solutions from previous studies.

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.

Elastic turbulence in entangled semi-dilute DNA solutions measured with optical coherence tomography velocimetry.

PubMed

Malm, A V; Waigh, T A

2017-04-26

The flow instabilities of solutions of high molecular weight DNA in the entangled semi-dilute concentration regime were investigated using optical coherence tomography velocimetry, a technique that provides high spatial (probe volumes of 3.4 pL) and temporal resolution (sub μs) information on the flow behaviour of complex fluids in a rheometer. The velocity profiles of the opaque DNA solutions (high and low salt) were measured as a function of the distance across the gap of a parallel plate rheometer, and their evolution over time was measured. At lower DNA concentrations and low shear rates, the velocity fluctuations were well described by Gaussian functions and the velocity gradient was uniform across the rheometer gap, which is expected for Newtonian flows. As the DNA concentration and shear rate were increased there was a stable wall slip regime followed by an evolving wall slip regime, which is finally followed by the onset of elastic turbulence. Strain localization (shear banding) is observed on the boundaries of the flows at intermediate shear rates, but decreases in the high shear elastic turbulence regime, where bulk strain localization occurs. A dynamic phase diagram for non-linear flow was created to describe the different behaviours.

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.

Impact of oral Lactobacillus acidophilus gavage on rooster seminal and cloacal Lactobacilli concentrations.

PubMed

Kiess, A S; Hirai, J H; Triplett, M D; Parker, H M; McDaniel, C D

2016-08-01

The use of antibiotics in poultry is being heavily scrutinized, therefore alternatives such as probiotics are being investigated. Lactobacilli spp. are a commonly used bacteria in formulating probiotics, and the addition of Lactobacilli to broiler diets has demonstrated increased growth rates, stimulated immune systems, and reduced pathogen loads in the gastro-intestinal tract ( GI: ) tract. However, previous research has shown that when rooster semen is directly exposed to Lactobacillus acidophilus (L. acidophilus) sperm quality is reduced. Therefore, the objective of the current study was to determine if oral administration of L. acidophilus increases the concentration of Lactobacilli in semen as well as the cloaca. A total of 30 roosters were used: 15 roosters were gavaged with 1X PBS (Control) and 15 roosters were gavaged with 10(7) cfu/mL of L. acidophilus (Treated). All roosters were gavaged for 14 consecutive days. Semen was collected on a 3 d interval, and cloacal swabs were collected on a 2 d interval, beginning on the first day prior to oral administration. Semen and cloacal swabs were serial diluted, and 100 μL of each dilution was then plated on Man, Rogosa, Sharpe ( MRS: ) agar plates. All plates were incubated for 48 h at 37°C under anaerobic conditions and counted. All Lactobacilli counts were first log transformed, then log transformed (day 0) pre-counts were subtracted from the log transformed day counts providing log differences for the analysis. Seminal Lactobacilli counts were not altered by treatments. However, the main effect of treatment (P = 0.026) for cloacal counts indicated that roosters gavaged with Lactobacilli yielded higher counts than the controls. Additionally, cloaca samples also demonstrated a treatment by day interaction trend (P = 0.082), where Lactobacilli was higher in the L. acidophilus gavaged roosters than the controls only on days 3, 5, 13, and 15. In conclusion, the addition of L. acidophilus to the male breeder diet over extended periods may increase concentrations of Lactobacilli in the cloaca even higher than the concentrations observed in this study. If Lactobacilli reaches high enough concentrations in the cloaca, then sperm quality may be impacted which could lead to poor fertility within the breeder flock. © 2016 Poultry Science Association Inc.

Reconstruction of transient vibration and sound radiation of an impacted plate using time domain plane wave superposition method

NASA Astrophysics Data System (ADS)

Geng, Lin; Zhang, Xiao-Zheng; Bi, Chuan-Xing

2015-05-01

Time domain plane wave superposition method is extended to reconstruct the transient pressure field radiated by an impacted plate and the normal acceleration of the plate. In the extended method, the pressure measured on the hologram plane is expressed as a superposition of time convolutions between the time-wavenumber normal acceleration spectrum on a virtual source plane and the time domain propagation kernel relating the pressure on the hologram plane to the normal acceleration spectrum on the virtual source plane. By performing an inverse operation, the normal acceleration spectrum on the virtual source plane can be obtained by an iterative solving process, and then taken as the input to reconstruct the whole pressure field and the normal acceleration of the plate. An experiment of a clamped rectangular steel plate impacted by a steel ball is presented. The experimental results demonstrate that the extended method is effective in visualizing the transient vibration and sound radiation of an impacted plate in both time and space domains, thus providing the important information for overall understanding the vibration and sound radiation of the plate.

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.

Studies on prevalence of Strongyloides infection in Holambra and Maceió, Brazil, by the agar plate faecal culture method.

PubMed

Kobayashi, J; Hasegawa, H; Soares, E C; Toma, H; Dacal, A R; Brito, M C; Yamanaka, A; Foli, A A; Sato, Y

1996-01-01

Prevalence of Strongyloides stercoralis infection in three areas of Brazil was surveyed by a recently developed faecal culture method (an agar plate culture). The Strongyloides infection was confirmed in 11.3% of 432 subjects examined. The diagnostic efficacy of the agar plate culture was as high as 93.9% compared to only 28.5% and 26.5% by the Harada-Mori filter paper culture and faecal concentration methods, when faecal samples were examined simultaneously by these three methods. Among the 49 positive samples, about 60% were confirmed to be positive only by the agar plate culture. These results indicate that the agar plate culture is a sensitive new tool for the correct diagnosis of chronic Strongyloides infection.

Plate equations for piezoelectrically actuated flexural mode ultrasound transducers.

PubMed

Perçin, Gökhan

2003-01-01

This paper considers variational methods to derive two-dimensional plate equations for piezoelectrically actuated flexural mode ultrasound transducers. In the absence of analytical expressions for the equivalent circuit parameters of a flexural mode transducer, it is difficult to calculate its optimal parameters and dimensions, and to choose suitable materials. The influence of coupling between flexural and extensional deformation, and coupling between the structure and the acoustic volume on the dynamic response of piezoelectrically actuated flexural mode transducer is analyzed using variational methods. Variational methods are applied to derive two-dimensional plate equations for the transducer, and to calculate the coupled electromechanical field variables. In these methods, the variations across the thickness direction vanish by using the stress resultants. Thus, two-dimensional plate equations for a stepwise laminated circular plate are obtained.

Rapid planar chromatographic analysis of 25 water-soluble dyes used as food additives.

PubMed

Morlock, Gertrud E; Oellig, Claudia

2009-01-01

A rapid planar chromatographic method for identification and quantification of 25 water-soluble dyes in food was developed. In a horizontal developing chamber, the chromatographic separation on silica gel 60F254 high-performance thin-layer chromatography plates took 12 min for 40 runs in parallel, using 8 mL ethyl acetate-methanol-water-acetic acid (65 + 23 + 11 + 1, v/v/v/v) mobile phase up to a migration distance of 50 mm. However, the total analysis time, inclusive of application and evaluation, took 60 min for 40 runs. Thus, the overall time/run can be calculated as 1.5 min with a solvent consumption of 200 microL. A sample throughput of 1000 runs/8 h day can be reached by switching between the working stations (application, development, and evaluation) in a 20 min interval, which triples the analysis throughput. Densitometry was performed by absorption measurement using the multiwavelength scan mode in the UV and visible ranges. Repeatabilities [relative standard deviation (RSD), 4 determinations] at the first or second calibration level showed precisions of mostly < or = 2.7%, ranging between 0.2 and 5.2%. Correlation coefficient values (R > or = 0.9987) and RSD values (< or = 4.2%) of the calibration curves were highly satisfactory using classical quantification. However, digital evaluation of the plate image was also used for quantification, which resulted in RSD values of the calibration curves of mostly < or = 3.0%, except for two < or = 6.0%. The method was applied for the analysis of some energy drinks and bakery ink formulations, directly applied after dilution. By recording of absorbance spectra in the visible range, the identities of the dyes found in the samples were ascertained by comparison with the respective standard bands (correlation coefficients > or = 0.9996). If necessary for confirmation, online mass spectra were recorded within a minute.

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.…

Congruency of scapula locking plates: implications for implant design.

PubMed

Park, Andrew Y; DiStefano, James G; Nguyen, Thuc-Quyen; Buckley, Jenni M; Montgomery, William H; Grimsrud, Chris D

2012-04-01

We conducted a study to evaluate the congruency of fit of current scapular plate designs. Three-dimensional image-processing and -analysis software, and computed tomography scans of 12 cadaveric scapulae were used to generate 3 measurements: mean distance from plate to bone, maximum distance, and percentage of plate surface within 2 mm of bone. These measurements were used to quantify congruency. The scapular spine plate had the most congruent fit in all 3 measured variables. The lateral border and glenoid plates performed statistically as well as the scapular spine plate in at least 1 of the measured variables. The medial border plate had the least optimal measurements in all 3 variables. With locking-plate technology used in a wide variety of anatomical locations, the locking scapula plate system can allow for a fixed-angle construct in this region. Our study results showed that the scapular spine, glenoid, and lateral border plates are adequate in terms of congruency. However, design improvements may be necessary for the medial border plate. In addition, we describe a novel method for quantifying hardware congruency, a method that can be applied to any anatomical location.

Three-Phase Coexistence in Colloidal Rod-Plate Mixtures.

PubMed

Woolston, Phillip; van Duijneveldt, Jeroen S

2015-09-01

Aqueous suspensions of clay particles, such as montmorillonite (MMT) platelets and sepiolite (Sep) rods, tend to form gels at concentrations around 1 vol %. For Sep rods, adsorbing sodium polyacrylate to the surface allows for an isotropic-nematic phase separation to be seen instead. Here, MMT is added to such Sep suspensions, resulting in a complex phase behavior. Across a range of clay concentrations, separation into three phases is observed: a lower, nematic phase dominated by Sep rods, a MMT-rich middle layer, which is weakly birefringent and probably a gel, and a dilute top phase. Analysis of phase volumes suggests that the middle layer may contain as much as 6 vol % MMT.

A 2D Daubechies finite wavelet domain method for transient wave response analysis in shear deformable laminated composite plates

NASA Astrophysics Data System (ADS)

Nastos, C. V.; Theodosiou, T. C.; Rekatsinas, C. S.; Saravanos, D. A.

2018-03-01

An efficient numerical method is developed for the simulation of dynamic response and the prediction of the wave propagation in composite plate structures. The method is termed finite wavelet domain method and takes advantage of the outstanding properties of compactly supported 2D Daubechies wavelet scaling functions for the spatial interpolation of displacements in a finite domain of a plate structure. The development of the 2D wavelet element, based on the first order shear deformation laminated plate theory is described and equivalent stiffness, mass matrices and force vectors are calculated and synthesized in the wavelet domain. The transient response is predicted using the explicit central difference time integration scheme. Numerical results for the simulation of wave propagation in isotropic, quasi-isotropic and cross-ply laminated plates are presented and demonstrate the high spatial convergence and problem size reduction obtained by the present method.

Biomechanical characteristics of fixation methods for floating pubic symphysis.

PubMed

Song, Wenhao; Zhou, Dongsheng; He, Yu

2017-03-07

Floating pubic symphysis (FPS) is a relatively rare injury caused by high-energy mechanisms. There are several fixation methods used to treat FPS, including external fixation, subcutaneous fixation, internal fixation, and percutaneous cannulated screw fixation. To choose the appropriate fixation, it is necessary to study the biomechanical performance of these different methods. The goal of this study was to compare the biomechanical characteristics of six methods by finite element analysis. A three-dimensional finite element model of FPS was simulated. Six methods were used in the FPS model, including external fixation (Ext), subcutaneous rod fixation (Sub-rod), subcutaneous plate fixation (Sub-plate), superior pectineal plate fixation (Int-sup), infrapectineal plate fixation (Int-ifa), and cannulated screw fixation (Int-scr). Compressive and rotational loads were then applied in all models. Biomechanical characteristics that were recorded and analyzed included construct stiffness, micromotion of the fracture gaps, von Mises stress, and stress distribution. The construct stiffness of the anterior pelvic ring was decreased dramatically when FPS occurred. Compressive stiffness was restored by the three internal fixation and Sub-rod methods. Unfortunately, rotational stiffness was not restored satisfactorily by the six methods. For micromotion of the fracture gaps, the displacement was reduced significantly by the Int-sup and Int-ifa methods under compression. The internal fixation methods and Sub-plate method performed well under rotation. The maximum von Mises stress of the implants was not large. For the plate-screw system, the maximum von Mises stress occurred over the region of the fracture and plate-screw joints. The maximum von Mises stress appeared on the rod-screw and screw-bone interfaces for the rod-screw system. The present study showed the biomechanical advantages of internal fixation methods for FPS from a finite element view. Superior stabilization of the anterior pelvic ring and fracture gaps was obtained by internal fixation. Subcutaneous fixation had satisfactory outcomes as well. Sub-rod fixation offered good anti-compression, while the Sub-plate fixation provided favorable anti-rotational capacity.

Evaluation of the methods for enumerating coliform bacteria from water samples using precise reference standards.

PubMed

Wohlsen, T; Bates, J; Vesey, G; Robinson, W A; Katouli, M

2006-04-01

To use BioBall cultures as a precise reference standard to evaluate methods for enumeration of Escherichia coli and other coliform bacteria in water samples. Eight methods were evaluated including membrane filtration, standard plate count (pour and spread plate methods), defined substrate technology methods (Colilert and Colisure), the most probable number method and the Petrifilm disposable plate method. Escherichia coli and Enterobacter aerogenes BioBall cultures containing 30 organisms each were used. All tests were performed using 10 replicates. The mean recovery of both bacteria varied with the different methods employed. The best and most consistent results were obtained with Petrifilm and the pour plate method. Other methods either yielded a low recovery or showed significantly high variability between replicates. The BioBall is a very suitable quality control tool for evaluating the efficiency of methods for bacterial enumeration in water samples.

Classification and treatment of periprosthetic supracondylar femur fractures.

PubMed

Ricci, William

2013-02-01

Locked plating and retrograde nailing are two accepted methods for treatment of periprosthetic distal femur fractures. Each has relative benefits and potential pitfalls. Appropriate patient selection and knowledge of the specific femoral component geometry are required to optimally choose between these two methods. Locked plating may be applied to most periprosthetic distal femur fractures. The fracture pattern, simple or comminuted, will dictate the specific plating technique, compression plating or bridge plating. Nailing requires an open intercondylar box and a distal fragment of enough size to allow interlocking. With proper patient selection and proper techniques, good results can be obtained with either method. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

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

Elastic guided waves in a layered plate with rectangular cross section.

PubMed

Mukdadi, O M; Desai, Y M; Datta, S K; Shah, A H; Niklasson, A J

2002-11-01

Guided waves in a layered elastic plate of rectangular cross section (finite width and thickness) has been studied in this paper. A semianalytical finite element method in which the deformation of the cross section is modeled by two-dimensional finite elements and analytical representation of propagating waves along the length of the plate has been used. The method is applicable to arbitrary number of layers and general anisotropic material properties of each layer, and is similar to the stiffness method used earlier to study guided waves in a laminated composite plate of infinite width. Numerical results showing the effect of varying the width of the plate on the dispersion of guided waves are presented and are compared with those for an infinite plate. In addition, effect of thin anisotropic coating or interface layers on the guided waves is investigated.

A Modified Kirchhoff plate theory for Free Vibration analysis of functionally graded material plates using meshfree method

NASA Astrophysics Data System (ADS)

Nguyen Van Do, Vuong

2018-04-01

In this paper, a modified Kirchhoff theory is presented for free vibration analyses of functionally graded material (FGM) plate based on modified radial point interpolation method (RPIM). The shear deformation effects are taken account into modified theory to ignore the locking phenomenon of thin plates. Due to the proposed refined plate theory, the number of independent unknowns reduces one variable and exists with four degrees of freedom per node. The simulated free vibration results employed by the modified RPIM are compared with the other analytical solutions to verify the effectiveness and the accuracy of the developed mesh-free method. Detail parametric studies of the proposed method are then conducted including the effectiveness of thickness ratio, boundary condition and material inhomogeneity on the sample problems of square plates. Results illustrated that the modified mesh-free RPIM can effectively predict the numerical calculation as compared to the exact solutions. The obtained numerical results are indicated that the proposed method are stable and well accurate prediction to evaluate with other published analyses.

Development of a Titanium Plate for Mandibular Angle Fractures with a Bone Defect in the Lower Border: Finite Element Analysis and Mechanical Test

PubMed Central

Goulart, Douglas Rangel; Kemmoku, Daniel Takanori; Noritomi, Pedro Yoshito

2015-01-01

ABSTRACT Objectives The aim of the present study was to develop a plate to treat mandibular angle fractures using the finite element method and mechanical testing. Material and Methods A three-dimensional model of a fractured mandible was generated using Rhinoceros 4.0 software. The models were exported to ANSYS®, in which a static application of displacement (3 mm) was performed in the first molar region. Three groups were assessed according to the method of internal fixation (2 mm system): two non-locking plates; two locking plates and a new design locking plate. The computational model was transferred to an in vitro experiment with polyurethane mandibles. Each group contained five samples and was subjected to a linear loading test in a universal testing machine. Results A balanced distribution of stress was associated with the new plate design. This plate modified the mechanical behavior of the fractured region, with less displacement between the fractured segments. In the mechanical test, the group with two locking plates exhibited greater resistance to the 3 mm displacement, with a statistically significant difference when compared with the new plate group (ANOVA, P = 0.016). Conclusions The new plate exhibited a more balanced distribution of stress. However, the group with two locking plates exhibited greater mechanical resistance. PMID:26539287

Alloy vapor deposition using ion plating and flash evaporation

NASA Technical Reports Server (NTRS)

Spalvins, T.

1971-01-01

Method extends scope of ion plating technique to include deposition of alloy films without changing composition of plating alloy. Coatings flow with specimen material without chipping or peeling. Technique is most effective vacuum deposition method for depositing alloys for strong and lasting adherence.

The complex variable reproducing kernel particle method for bending problems of thin plates on elastic foundations

NASA Astrophysics Data System (ADS)

Chen, L.; Cheng, Y. M.

2018-07-01

In this paper, the complex variable reproducing kernel particle method (CVRKPM) for solving the bending problems of isotropic thin plates on elastic foundations is presented. In CVRKPM, one-dimensional basis function is used to obtain the shape function of a two-dimensional problem. CVRKPM is used to form the approximation function of the deflection of the thin plates resting on elastic foundation, the Galerkin weak form of thin plates on elastic foundation is employed to obtain the discretized system equations, the penalty method is used to apply the essential boundary conditions, and Winkler and Pasternak foundation models are used to consider the interface pressure between the plate and the foundation. Then the corresponding formulae of CVRKPM for thin plates on elastic foundations are presented in detail. Several numerical examples are given to discuss the efficiency and accuracy of CVRKPM in this paper, and the corresponding advantages of the present method are shown.

Method and apparatus for close packing of nuclear fuel assemblies

DOEpatents

Newman, Darrell F.

1993-01-01

The apparatus of the present invention is a plate of neutron absorbing material. The plate may have a releasable locking feature permitting the plate to be secured within a nuclear fuel assembly between nuclear fuel rods during storage or transportation then removed for further use or destruction. The method of the present invention has the step of placing a plate of neutron absorbing material between nuclear fuel rods within a nuclear fuel assembly, preferably between the two outermost columns of nuclear fuel rods. Additionally, the plate may be releasably locked in place.

Method and apparatus for close packing of nuclear fuel assemblies

DOEpatents

Newman, D.F.

1993-03-30

The apparatus of the present invention is a plate of neutron absorbing material. The plate may have a releasable locking feature permitting the plate to be secured within a nuclear fuel assembly between nuclear fuel rods during storage or transportation then removed for further use or destruction. The method of the present invention has the step of placing a plate of neutron absorbing material between nuclear fuel rods within a nuclear fuel assembly, preferably between the two outermost columns of nuclear fuel rods. Additionally, the plate may be releasably locked in place.

Zone plate method for electronic holographic display using resolution redistribution technique.

PubMed

Takaki, Yasuhiro; Nakamura, Junya

2011-07-18

The resolution redistribution (RR) technique can increase the horizontal viewing-zone angle and screen size of electronic holographic display. The present study developed a zone plate method that would reduce hologram calculation time for the RR technique. This method enables calculation of an image displayed on a spatial light modulator by performing additions of the zone plates, while the previous calculation method required performing the Fourier transform twice. The derivation and modeling of the zone plate are shown. In addition, the look-up table approach was introduced for further reduction in computation time. Experimental verification using a holographic display module based on the RR technique is presented.

Investigation on filter method for smoothing spiral phase plate

NASA Astrophysics Data System (ADS)

Zhang, Yuanhang; Wen, Shenglin; Luo, Zijian; Tang, Caixue; Yan, Hao; Yang, Chunlin; Liu, Mincai; Zhang, Qinghua; Wang, Jian

2018-03-01

Spiral phase plate (SPP) for generating vortex hollow beams has high efficiency in various applications. However, it is difficult to obtain an ideal spiral phase plate because of its continuous-varying helical phase and discontinued phase step. This paper describes the demonstration of continuous spiral phase plate using filter methods. The numerical simulations indicate that different filter method including spatial domain filter, frequency domain filter has unique impact on surface topography of SPP and optical vortex characteristics. The experimental results reveal that the spatial Gaussian filter method for smoothing SPP is suitable for Computer Controlled Optical Surfacing (CCOS) technique and obtains good optical properties.

Diabetes Diet, Eating, & Physical Activity

MedlinePlus

... for pregnancy if you have diabetes. Meal plan methods Two common ways to help you plan how ... about the method that’s best for you. Plate method The plate method helps you control your portion ...

Multipotent Mesenchymal Stromal Stem Cell Expansion by Plating Whole Bone Marrow at a Low Cellular Density: A More Advantageous Method for Clinical Use

PubMed Central

Mareschi, Katia; Rustichelli, Deborah; Calabrese, Roberto; Gunetti, Monica; Sanavio, Fiorella; Castiglia, Sara; Risso, Alessandra; Ferrero, Ivana; Tarella, Corrado; Fagioli, Franca

2012-01-01

Mesenchymal stem cells (MSCs) are a promising source for cell therapy due to their pluripotency and immunomodulant proprieties. As the identification of “optimal” conditions is important to identify a standard procedure for clinical use. Percoll, Ficoll and whole bone marrow directly plated were tested from the same sample as separation methods. The cells were seeded at the following densities: 100 000, 10 000, 1000, 100, 10 cells/cm2. After reaching confluence, the cells were detached, pooled and re-plated at 1000, 500, 100, and 10 cells/cm2. Statistical analyses were performed. Cumulative Population Doublings (PD) did not show significant differences for the separation methods and seeding densities but only for the plating density. Some small quantity samples plated in T25 flasks at plating densities of 10 and 100 cells/cm2 did not produce any expansion. However, directly plated whole bone marrow resulted in a more advantageous method in terms of CFU-F number, cellular growth and minimal manipulation. No differences were observed in terms of gross morphology, differentiation potential or immunophenotype. These data suggest that plating whole bone marrow at a low cellular density may represent a good procedure for MSC expansion for clinical use. PMID:23715383

Chromatic characterization of ion-exchanged glass binary phase plates for mode-division multiplexing.

PubMed

Prieto-Blanco, Xesús; Montero-Orille, Carlos; Moreno, Vicente; Mateo, Eduardo F; Liñares, Jesús

2015-04-10

Mode-division multiplexing (MDM) in few-mode fibers is regarded as a promising candidate to increase optical network capacity. A fundamental element for MDM is a modal transformer to LP modes which can be implemented in a free-space basis by using multiregion phase plates, that is, LP plates. Likewise, several wavelengths have to be used due to wavelength multiplexing purposes, optical amplification tasks, and so on. In this work we show that efficient monolithic binary phase plates for different wavelengths can be fabricated by ion-exchange in glass and used for MDM tasks. We introduce an optical characterization method of the chromatic properties of such phase plates which combines the inverse Wentzel-Kramers-Brillouin (IWKB) together with Mach-Zehnder and Michelson-based interferometric techniques. The interferometric method provides a measurement of the phase step for several wavelengths, which characterizes the chromatic properties of the phase plate. Consequently, it is shown that the IWKB method allows us to design and characterize the phase plates in an easy and fast way.

Measurement of large steel plates based on linear scan structured light scanning

NASA Astrophysics Data System (ADS)

Xiao, Zhitao; Li, Yaru; Lei, Geng; Xi, Jiangtao

2018-01-01

A measuring method based on linear structured light scanning is proposed to achieve the accurate measurement of the complex internal shape of large steel plates. Firstly, by using a calibration plate with round marks, an improved line scanning calibration method is designed. The internal and external parameters of camera are determined through the calibration method. Secondly, the images of steel plates are acquired by line scan camera. Then the Canny edge detection method is used to extract approximate contours of the steel plate images, the Gauss fitting algorithm is used to extract the sub-pixel edges of the steel plate contours. Thirdly, for the problem of inaccurate restoration of contour size, by measuring the distance between adjacent points in the grid of known dimensions, the horizontal and vertical error curves of the images are obtained. Finally, these horizontal and vertical error curves can be used to correct the contours of steel plates, and then combined with the calibration parameters of internal and external, the size of these contours can be calculated. The experiments results demonstrate that the proposed method can achieve the error of 1 mm/m in 1.2m×2.6m field of view, which has satisfied the demands of industrial measurement.

Multidimensional Normalization to Minimize Plate Effects of Suspension Bead Array Data.

PubMed

Hong, Mun-Gwan; Lee, Woojoo; Nilsson, Peter; Pawitan, Yudi; Schwenk, Jochen M

2016-10-07

Enhanced by the growing number of biobanks, biomarker studies can now be performed with reasonable statistical power by using large sets of samples. Antibody-based proteomics by means of suspension bead arrays offers one attractive approach to analyze serum, plasma, or CSF samples for such studies in microtiter plates. To expand measurements beyond single batches, with either 96 or 384 samples per plate, suitable normalization methods are required to minimize the variation between plates. Here we propose two normalization approaches utilizing MA coordinates. The multidimensional MA (multi-MA) and MA-loess both consider all samples of a microtiter plate per suspension bead array assay and thus do not require any external reference samples. We demonstrate the performance of the two MA normalization methods with data obtained from the analysis of 384 samples including both serum and plasma. Samples were randomized across 96-well sample plates, processed, and analyzed in assay plates, respectively. Using principal component analysis (PCA), we could show that plate-wise clusters found in the first two components were eliminated by multi-MA normalization as compared with other normalization methods. Furthermore, we studied the correlation profiles between random pairs of antibodies and found that both MA normalization methods substantially reduced the inflated correlation introduced by plate effects. Normalization approaches using multi-MA and MA-loess minimized batch effects arising from the analysis of several assay plates with antibody suspension bead arrays. In a simulated biomarker study, multi-MA restored associations lost due to plate effects. Our normalization approaches, which are available as R package MDimNormn, could also be useful in studies using other types of high-throughput assay data.

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.

Optimum structural design with plate bending elements - A survey

NASA Technical Reports Server (NTRS)

Haftka, R. T.; Prasad, B.

1981-01-01

A survey is presented of recently published papers in the field of optimum structural design of plates, largely with respect to the minimum-weight design of plates subject to such constraints as fundamental frequency maximization. It is shown that, due to the availability of powerful computers, the trend in optimum plate design is away from methods tailored to specific geometry and loads and toward methods that can be easily programmed for any kind of plate, such as finite element methods. A corresponding shift is seen in optimization from variational techniques to numerical optimization algorithms. Among the topics covered are fully stressed design and optimality criteria, mathematical programming, smooth and ribbed designs, design against plastic collapse, buckling constraints, and vibration constraints.

A correlation method to predict the surface pressure distribution on an infinite plate from which a jet is issuing. [effects of a lifting jet

NASA Technical Reports Server (NTRS)

Perkins, S. C., Jr.; Menhall, M. R.

1978-01-01

A correlation method to predict pressures induced on an infinite plate by a jet issuing from the plate into a subsonic free stream was developed. The complete method consists of an analytical method which models the blockage and entrainment properties of the jet and a correlation which accounts for the effects of separation. The method was developed for jet velocity ratios up to ten and for radial distances up to five diameters from the jet. Correlation curves and data comparisons are presented for jets issuing normally from a flat plate with velocity ratios one to twelve. Also, a list of references which deal with jets in a crossflow is presented.

Characterization of indoor bioaerosols from a hospital ward in a tropical setting.

PubMed

Sudharsanam, S; Swaminathan, S; Ramalingam, A; Thangavel, G; Annamalai, R; Steinberg, R; Balakrishnan, K; Srikanth, P

2012-06-01

Study was conducted to assess whether temporal variation exists in airborne microbial concentrations of a hospital ward (west-Chennai, India) using active and passive methods, and characterise the microorganisms. Air samples (duplicates) were collected simultaneously using exposed-plate, impingement (BioSampler) and filtration (personal sampling filter cassette loaded with gelatin filter) methods over different periods of the year. Bacterial plates were incubated at 37°C and observed for growth after 48h; fungal plates were incubated at 25°C and 37°C and observed upto 7 days. Microorganisms were identified using standard microbiological procedures. Microbial loads were found to vary with the sampling method. Concentrations of bacteria were higher (exposed-plate: 45-150 CFU/plate; impingement: 1.12E+03-1.6856E+05 CFU/m(3); filtration: 3.788E+03-1.91111E+05 CFU/m(3)) than fungi (exposed-plate: 0-13 CFU/plate; impingement: 0-3.547E+03 CFU/m(3); filtration: 0-1.515E+04 CFU/ m(3)). Coagulase-negative Staphylococci and Micrococci were the predominant Gram-positive cocci in active and passive samples. Enterobacter and Pseudomonas were the predominant Gram-negative bacilli. Among fungi, Aspergillus niger was isolated throughout the year. There was no significant temporal variation in airborne microbial loads irrespective of methods. Exposed-plate method was found to capture microorganisms efficiently with little variation in duplicate samples, suggesting its use in hospitals for preliminary assessment of indoor air quality and determine pathogenic microorganisms due to particle fall-out.

The analysis of the accuracy of the wheel alignment inspection method on the side-slip plate stand

NASA Astrophysics Data System (ADS)

Gajek, A.; Strzępek, P.

2016-09-01

The article presents the theoretical basis and the results of the examination of the wheel alignment inspection method on the slide slip plate stand. It is obligatory test during periodic technical inspection of the vehicle. The measurement is executed in the dynamic conditions. The dependence between the lateral displacement of the plate and toe-in of the tested wheels has been shown. If the diameter of the wheel rim is known then the value of the toe-in can be calculated. The comparison of the toe-in measurements on the plate stand and on the four heads device for the wheel alignment inspection has been carried out. The accuracy of the measurements and the influence of the conditions of the tests on the plate stand (the way of passing through the plate) were estimated. The conclusions about the accuracy of this method are presented.

System and method for optically locating microchannel positions

DOEpatents

Brewer, Laurence R.; Kimbrough, Joseph; Balch, Joseph; Davidson, J. Courtney

2001-01-01

A system and method is disclosed for optically locating a microchannel position. A laser source generates a primary laser beam which is directed at a microchannel plate. The microchannel plates include microchannels at various locations. A back-reflectance beam detector receives a back-reflected beam from the plate. The back-reflected beam is generated when the primary beam reflects off of the plate. A photodiode circuit generates a trigger signal when the back-reflected beam exceeds a predetermined threshold, indicating a presence of the microchannel. The method of the present invention includes the steps of generating a primary beam, directing the primary beam to a plate containing a microchannel, receiving from the plate a back-reflected beam generated in response to the primary beam, and generating a trigger signal when the back-reflected beam exceeds a predetermined threshold which corresponds to a presence of the microchannel.

Apparatus for in-situ nondestructive measurement of Young's modulus of plate structures

NASA Technical Reports Server (NTRS)

Huang, Jerry Qixin (Inventor); Perez, Robert J. (Inventor); DeLangis, Leo M. (Inventor)

2005-01-01

A method and apparatus for determining stiffness of a plate-like structure including a monolithic or composite laminate plate entails disposing a device for generating an acoustical pulse against a surface of the plate and disposing a detecting device against the same surface spaced a known distance from the pulse-generating device, and using the pulse-generating device to emit a pulse so as to create an extensional wave in the plate. The detecting device is used to determine a time of flight of the wave over the known distance, and the wave velocity is calculated. A Young's modulus of the plate is determined by a processor based on the wave velocity. Methods and apparatus for evaluating both isotropic plates and anisotropic laminates are disclosed.

Nondestructive test method accurately sorts mixed bolts

NASA Technical Reports Server (NTRS)

Dezeih, C. J.

1966-01-01

Neutron activation analysis method sorts copper plated steel bolts from nickel plated steel bolts. Copper and nickel plated steel bolt specimens of the same configuration are irradiated with thermal neutrons in a test reactor for a short time. After thermal neutron irradiation, the bolts are analyzed using scintillation energy readout equipment.

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

Development efforts to improve curved-channel microchannel plates

NASA Technical Reports Server (NTRS)

Corbett, M. B.; Feller, W. B.; Laprade, B. N.; Cochran, R.; Bybee, R.; Danks, A.; Joseph, C.

1993-01-01

Curved-channel microchannel plate (C-plate) improvements resulting from an ongoing NASA STIS microchannel plate (MCP) development program are described. Performance limitations of previous C-plates led to a development program in support of the STIS MAMA UV photon counter, a second generation instrument on the Hubble Space Telescope. C-plate gain, quantum detection efficiency, dark noise, and imaging distortion, which are influenced by channel curvature non-uniformities, have all been improved through use of a new centrifuge fabrication technique. This technique will be described, along with efforts to improve older, more conventional shearing methods. Process optimization methods used to attain targeted C-plate performance goals will be briefly characterized. Newly developed diagnostic measurement techniques to study image distortion, gain uniformity, input bias angle, channel curvature, and ion feedback, will be described. Performance characteristics and initial test results of the improved C-plates will be reported. Future work and applications will also be discussed.

Actively controlling coolant-cooled cold plate configuration

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

Chainer, Timothy J.; Parida, Pritish R.

A method is provided to facilitate active control of thermal and fluid dynamic performance of a coolant-cooled cold plate. The method includes: monitoring a variable associated with at least one of the coolant-cooled cold plate or one or more electronic components being cooled by the cold plate; and dynamically varying, based on the monitored variable, a physical configuration of the cold plate. By dynamically varying the physical configuration, the thermal and fluid dynamic performance of the cold plate are adjusted to, for example, optimally cool the one or more electronic components, and at the same time, reduce cooling power consumptionmore » used in cooling the electronic component(s). The physical configuration can be adjusted by providing one or more adjustable plates within the coolant-cooled cold plate, the positioning of which may be adjusted based on the monitored variable.« less

Characterization of E 471 food emulsifiers by high-performance thin-layer chromatography-fluorescence detection.

PubMed

Oellig, Claudia; Brändle, Klara; Schwack, Wolfgang

2018-07-13

Mono- and diacylglycerol (MAG and DAG) emulsifiers, also known as food additive E 471, are widely used to adjust techno-functional properties in various foods. Besides MAGs and DAGs, E 471 emulsifiers additionally comprise different amounts of triacylglycerols (TAGs) and free fatty acids (FFAs). MAGs, DAGs, TAGs and FFAs are generally determined by high-performance liquid chromatography (HPLC) or gas chromatography (GC) coupled to mass selective detection, analyzing the individual representatives of the lipid classes. In this work we present a rapid and sensitive method for the determination of MAGs, DAGs, TAGs and FFAs in E 471 emulsifiers by high-performance thin-layer chromatography with fluorescence detection (HPTLC-FLD), including a response factor system for quantitation. Samples were simply dissolved and diluted with t-butyl methyl ether before a two-fold development was performed on primuline pre-impregnated LiChrospher silica gel plates with diethyl ether and n-pentane/n-hexane/diethyl ether (52:20:28, v/v/v) as the mobile phases to 18 and 75 mm, respectively. For quantitation, the plate was scanned in the fluorescence mode at UV 366/>400 nm, when the cumulative signal for each lipid class was used. Calibration was done with 1,2-distearin and amounts of lipid classes were calculated with response factors and expressed as monostearin, distearin, tristearin and stearic acid. Limits of detection and quantitation were 1 and 4 ng/zone, respectively, for 1,2-distearin. Thus, the HPTLC-FLD approach represents a simple, rapid and convenient screening alternative to HPLC and GC analysis of the individual compounds. Visual detection additionally enables an easy characterization and the direct comparison of emulsifiers through the lipid class pattern, when utilized as a fingerprint. Copyright © 2018 Elsevier B.V. All rights reserved.

Inhibitory activity of a standardized elderberry liquid extract against clinically-relevant human respiratory bacterial pathogens and influenza A and B viruses

PubMed Central

2011-01-01

Background Black elderberries (Sambucus nigra L.) are well known as supportive agents against common cold and influenza. It is further known that bacterial super-infection during an influenza virus (IV) infection can lead to severe pneumonia. We have analyzed a standardized elderberry extract (Rubini, BerryPharma AG) for its antimicrobial and antiviral activity using the microtitre broth micro-dilution assay against three Gram-positive bacteria and one Gram-negative bacteria responsible for infections of the upper respiratory tract, as well as cell culture experiments for two different strains of influenza virus. Methods The antimicrobial activity of the elderberry extract was determined by bacterial growth experiments in liquid cultures using the extract at concentrations of 5%, 10%, 15% and 20%. The inhibitory effects were determined by plating the bacteria on agar plates. In addition, the inhibitory potential of the extract on the propagation of human pathogenic H5N1-type influenza A virus isolated from a patient and an influenza B virus strain was investigated using MTT and focus assays. Results For the first time, it was shown that a standardized elderberry liquid extract possesses antimicrobial activity against both Gram-positive bacteria of Streptococcus pyogenes and group C and G Streptococci, and the Gram-negative bacterium Branhamella catarrhalis in liquid cultures. The liquid extract also displays an inhibitory effect on the propagation of human pathogenic influenza viruses. Conclusion Rubini elderberry liquid extract is active against human pathogenic bacteria as well as influenza viruses. The activities shown suggest that additional and alternative approaches to combat infections might be provided by this natural product. PMID:21352539

Determination of optimal conditions for 5-methyl-benzotriazole biodegradation with activated sludge communities by dilution of the inoculum.

PubMed

Yuan, Heyang; Herzog, Bastian; Helmreich, Brigitte; Lemmer, Hilde; Müller, Elisabeth

2014-07-15

The aerobic biodegradation of 5-methyl-benzotriazole (5-TTri) was optimized using lab-scale setups and activated sludge communities (ASC) collected from three wastewater treatment plants (WWTP) MBR-MH, CAS-E and CAS-M being different in their treatment technologies. ASC inocula were diluted to rule out non-biodegrading species and incubated under two nutrient conditions: A) mineral salt media (MSM) and B) carbon and nitrogen supplied MSM giving MSM-CN. 5-TTri removal with the ASC ranged from 60% to 100% in only 10 days. 100 μL suspended biomass from the biodegrading setups was subsequently plated on solid media to eliminate possible activated sludge remnants. After growth occurred, mixed colonies were harvested and inoculated in fresh liquid MSM containing 20 mg L(-1) 5-TTri. These bacterial consortia showed good 5-TTri removal in MSM-CN rather than in MSM, indicating nutrient supply being required for efficient biodegradation. In addition, experiments with high 5-TTri concentrations ranging from 20 to 1,000 mg L(-1) were conducted in both, MSM and MSM-CN and the maximal 5-TTri removal capacity of the ASC evaluated. 50 mg L(-1) 5-TTri was still removed in both media whereas 100 mg L(-1) was solely removed in MSM-CN. 5-TTri biodegradation patterns also indicated that 5-TTri might be co-metabolized by microbial consortia. Furthermore, experiments with gradient-solid-media-plates showed 5-TTri to be inhibitory for the ASC in concentrations above 50 mg L(-1) and revealed the optimal conditions regarding carbon and nitrogen concentration and pH value for effective 5-TTri biodegradation by ASC. Nitrogen proved a crucial factor for enhancing organisms' biodegradation capacity with an optimal pH around 7 while carbon showed no such effect. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of the antimicrobial effect of super-oxidized water (Sterilox®) and sodium hypochlorite against Enterococcus faecalis in a bovine root canal model

PubMed Central

ROSSI-FEDELE, Giampiero; de FIGUEIREDO, José Antonio Poli; STEIER, Liviu; CANULLO, Luigi; STEIER, Gabriela; ROBERTS, Adam P.

2010-01-01

Ideally root canal irrigants should have, amongst other properties, antimicrobial action associated with a lack of toxicity against periapical tissues. Sodium hypochlorite (NaOCl) is a widely used root canal irrigant, however it has been shown to have a cytotoxic effect on vital tissue and therefore it is prudent to investigate alternative irrigants. Sterilox's Aquatine Alpha Electrolyte® belongs to the group of the super-oxidized waters; it consists of a mixture of oxidizing substances, and has been suggested to be used as root canal irrigant. Super-oxidized waters have been shown to provide efficient cleaning of root canal walls, and have been proposed to be used for the disinfection of medical equipment. Objective To compare the antimicrobial action against Enterococcus faecalis of NaOCl, Optident Sterilox Electrolyte Solution® and Sterilox's Aquatine Alpha Electrolyte® when used as irrigating solutions in a bovine root canal model. Methodology Root sections were prepared and inoculated with E. faecalis JH2-2. After 10 days of incubation the root canals were irrigated using one of three solutions (NaOCl, Optident Sterilox Electrolyte Solution® and Sterilox's Aquatine Alpha Electrolyte®) and subsequently sampled by grinding dentin using drills. The debris was placed in BHI broth and dilutions were plated onto fresh agar plates to quantify growth. Results Sodium hypochlorite was the only irrigant to eliminate all bacteria. When the dilutions were made, although NaOCl was still statistically superior, Sterilox's Aquatine Alpha Electrolyte® solution was superior to Optident Sterilox Electrolyte Solution®. Conclusion Under the conditions of this study Sterilox's Aquatine Alpha Electrolyte® appeared to have significantly more antimicrobial action compared to the Optident Sterilox Electrolyte Solution® alone, however NaOCl was the only solution able to consistently eradicate E. faecalis in the model. PMID:21085808

LabKey Server NAb: A tool for analyzing, visualizing and sharing results from neutralizing antibody assays

PubMed Central

2011-01-01

Background Multiple types of assays allow sensitive detection of virus-specific neutralizing antibodies. For example, the extent of antibody neutralization of HIV-1, SIV and SHIV can be measured in the TZM-bl cell line through the degree of luciferase reporter gene expression after infection. In the past, neutralization curves and titers for this standard assay have been calculated using an Excel macro. Updating all instances of such a macro with new techniques can be unwieldy and introduce non-uniformity across multi-lab teams. Using Excel also poses challenges in centrally storing, sharing and associating raw data files and results. Results We present LabKey Server's NAb tool for organizing, analyzing and securely sharing data, files and results for neutralizing antibody (NAb) assays, including the luciferase-based TZM-bl NAb assay. The customizable tool supports high-throughput experiments and includes a graphical plate template designer, allowing researchers to quickly adapt calculations to new plate layouts. The tool calculates the percent neutralization for each serum dilution based on luminescence measurements, fits a range of neutralization curves to titration results and uses these curves to estimate the neutralizing antibody titers for benchmark dilutions. Results, curve visualizations and raw data files are stored in a database and shared through a secure, web-based interface. NAb results can be integrated with other data sources based on sample identifiers. It is simple to make results public after publication by updating folder security settings. Conclusions Standardized tools for analyzing, archiving and sharing assay results can improve the reproducibility, comparability and reliability of results obtained across many labs. LabKey Server and its NAb tool are freely available as open source software at http://www.labkey.com under the Apache 2.0 license. Many members of the HIV research community can also access the LabKey Server NAb tool without installing the software by using the Atlas Science Portal (https://atlas.scharp.org). Atlas is an installation of LabKey Server. PMID:21619655

Conductivity fuel cell collector plate and method of fabrication

DOEpatents

Braun, James C.

2002-01-01

An improved method of manufacturing a PEM fuel cell collector plate is disclosed. During molding a highly conductive polymer composite is formed having a relatively high polymer concentration along its external surfaces. After molding the polymer rich layer is removed from the land areas by machining, grinding or similar process. This layer removal results in increased overall conductivity of the molded collector plate. The polymer rich surface remains in the collector plate channels, providing increased mechanical strength and other benefits to the channels. The improved method also permits greater mold cavity thickness providing a number of advantages during the molding process.

Fabricating cooled electronic system with liquid-cooled cold plate and thermal spreader

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

Chainer, Timothy J.; Graybill, David P.; Iyengar, Madhusudan K.

Methods are provided for facilitating cooling of an electronic component. The method includes providing a liquid-cooled cold plate and a thermal spreader associated with the cold plate. The cold plate includes multiple coolant-carrying channel sections extending within the cold plate, and a thermal conduction surface with a larger surface area than a surface area of the component to be cooled. The thermal spreader includes one or more heat pipes including multiple heat pipe sections. One or more heat pipe sections are partially aligned to a first region of the cold plate, that is, where aligned to the surface to bemore » cooled, and partially aligned to a second region of the cold plate, which is outside the first region. The one or more heat pipes facilitate distribution of heat from the electronic component to coolant-carrying channel sections of the cold plate located in the second region of the cold plate.« less

Fabricating cooled electronic system with liquid-cooled cold plate and thermal spreader

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

Chainer, Timothy J.; Graybill, David P.; Iyengar, Madhusudan K.

Methods are provided for facilitating cooling of an electronic component. The methods include providing a liquid-cooled cold plate and a thermal spreader associated with the cold plate. The cold plate includes multiple coolant-carrying channel sections extending within the cold plate, and a thermal conduction surface with a larger surface area than a surface area of the component to be cooled. The thermal spreader includes one or more heat pipes including multiple heat pipe sections. One or more heat pipe sections are partially aligned to a first region of the cold plate, that is, where aligned to the surface to bemore » cooled, and partially aligned to a second region of the cold plate, which is outside the first region. The one or more heat pipes facilitate distribution of heat from the electronic component to coolant-carrying channel sections of the cold plate located in the second region of the cold plate.« less

Optimization of Uranium Molecular Deposition for Alpha-Counting Sources

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

Monzo, Ellen; Parsons-Moss, Tashi; Genetti, Victoria

2016-12-12

Method development for molecular deposition of uranium onto aluminum 1100 plates was conducted with custom plating cells at Lawrence Livermore National Laboratory. The method development focused primarily on variation of electrode type, which was expected to directly influence plated sample homogeneity. Solid disc platinum and mesh platinum anodes were compared and data revealed that solid disc platinum anodes produced more homogenous uranium oxide films. However, the activity distribution also depended on the orientation of the platinum electrode relative to the aluminum cathode, starting current, and material composition of the plating cell. Experiments demonstrated these variables were difficult to control undermore » the conditions available. Variation of plating parameters among a series of ten deposited plates yielded variations up to 30% in deposition efficiency. Teflon particles were observed on samples plated in Teflon cells, which poses a problem for alpha activity measurements of the plates. Preliminary electropolishing and chemical polishing studies were also conducted on the aluminum 1100 cathode plates.« less

Solving the Problem of Linear Viscoelasticity for Piecewise-Homogeneous Anisotropic Plates

NASA Astrophysics Data System (ADS)

Kaloerov, S. A.; Koshkin, A. A.

2017-11-01

An approximate method for solving the problem of linear viscoelasticity for thin anisotropic plates subject to transverse bending is proposed. The method of small parameter is used to reduce the problem to a sequence of boundary problems of applied theory of bending of plates solved using complex potentials. The general form of complex potentials in approximations and the boundary conditions for determining them are obtained. Problems for a plate with elliptic elastic inclusions are solved as an example. The numerical results for a plate with one, two elliptical (circular), and linear inclusions are analyzed.

Comparison of effects of seven treatment methods for distal radius fracture on minimizing complex regional pain syndrome

PubMed Central

Sun, Tao

2016-01-01

Introduction Using network meta-analysis, we evaluated the adverse effects of the seven most common treatment methods, i.e., bridging external fixation, non-bridging external fixation, K-wire fixation, plaster fixation, dorsal plating, volar plating, and dorsal and volar plating, by their associated risk of developing complex regional pain syndrome (CRPS) in distal radius fracture (DRF) patients. Material and methods Following an exhaustive search of scientific literature databases for high quality studies, randomized controlled trials (RCTs) related to our study topic were screened and selected based on stringent predefined inclusion and exclusion criteria. Data extracted from the selected studies were used for statistical analyses using Stata 12.0 software. Results A total of 17 RCTs, including 1658 DRF patients, were enrolled in this network meta-analysis. Among the 1658 DRF patients, 452 received bridging external fixation, 525 received non-bridging external fixation, 154 received K-wire fixation, 84 received plaster fixation, 132 received dorsal plating, 123 received volar plating, and 188 received dorsal and volar plating. When compared to bridging external fixation patients, there was no marked difference in the CRPS risk in DRF patients receiving different treatments (all p > 0.05). However, the surface under the cumulative ranking curves (SUCRA) for plaster fixation (77.0%) and non-bridging external fixation (71.3%) were significantly higher compared with the other five methods. Conclusions Our findings suggest that compared with bridging external fixation, K-wire fixation, dorsal plating, volar plating, dorsal and volar plating, plaster fixation and non-bridging external fixation might be the better treatment methods to reduce the risk of CRPS in DRF patients. PMID:28144268

[Finite element study of maxillary Le Fort-I osteotomy with rigid internal fixation].

PubMed

Zhou, Jian; Sun, Geng-Lin; Wu, Wei; Xu, Chong-Tao; Wang, Peng-Lin

2010-05-01

To study the biomechanical characteristic of maxillary Le fort- I osteotomy with rigid internal fixation (RIF) , so as to choose best fixation method. The 3-dimensional finite element models of maxillary Le Fort-I osteotomy with 9 kinds of RIF methods were established. Then the models were divided into three groups to calculate the stress distribution of the maxilla and the displacement of bone segment under 3 kinds of occlusion condition. The fixation stability of the different RIF methods was evaluated. Under the incisor occlusion condition, the stress of the cranio maxillary complex transmits mainly along the nasal-maxillary buttress. Under the premolar and molar occlusion condition, the stress transmits along the alveolar process first, then turns to the nasal-maxillary and zygomatic-maxillary buttress. The focused stress position of the internal fixation system is at the connection between the screws and the plate and at the plate near the osteotomy line. Under the premolar occlusion condition, the displacement of bone segment with different RIF methods was (in a decreasing order) 0.396509 mm (with bio-absorbable plate), 0.148393 mm (with micro-plate ), 0.078436 mm (with mini-plate) in group 1; 0.188791 mm (fixing at the nasal-maxillary buttress), 0.121718 mm (fixing at the zygomatic-maxillary buttress), 0.078436 mm (fixing at the both buttress) in group 2; 0.091023 mm (with straight plate), 0.078436 mm (with L shape plate), 0.072450 mm (with Y shape plate), 0.065617 mm (with T shape plate) in group 3. The fixation stability of using the bio-absorbable plate in Le Fort-I osteotomy is less stable than using the titanium plate. Fixing at the zygomatic-maxillary buttress is more stable than at the naso-maxillary buttress. The fixation stability is different by using different shapes of plates.

A generalized plate method for estimating total aerobic microbial count.

PubMed

Ho, Kai Fai

2004-01-01

The plate method outlined in Chapter 61: Microbial Limit Tests of the U.S. Pharmacopeia (USP 61) provides very specific guidance for assessing total aerobic bioburden in pharmaceutical articles. This methodology, while comprehensive, lacks the flexibility to be useful in all situations. By studying the plate method as a special case within a more general family of assays, the effects of each parameter in the guidance can be understood. Using a mathematical model to describe the plate counting procedure, a statistical framework for making more definitive statements about total aerobic bioburden is developed. Such a framework allows the laboratory scientist to adjust the USP 61 methods to satisfy specific practical constraints. In particular, it is shown that the plate method can be conducted, albeit with stricter acceptance criteria, using a test specimen quantity that is smaller than the 10 g or 10 mL prescribed in the guidance. Finally, the interpretation of results proffered by the guidance is re-examined within this statistical framework and shown to be overly aggressive.

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.

Semi-automatic for ultrasonic measurement of texture

DOEpatents

Thompson, R. Bruce; Smith, John F.; Lee, Seung S.; Li, Yan

1990-02-13

A method for measuring texture of metal plates or sheets using non-destructive ultrasonic investigation includes measuring the velocity of ultrasonic energy waves in lower order plate modes in one or more directions, and measuring phase velocity dispersion of higher order modes of the plate or sheet if needed. Texture or preferred grain orientation can be derived from these measurements with improved reliability and accuracy. The method can be utilized in production on moving metal plate or sheet.

Semi-automatic for ultrasonic measurement of texture

DOEpatents

Thompson, R.B.; Smith, J.F.; Lee, S.S.; Li, Y.

1990-02-13

A method for measuring texture of metal plates or sheets using non-destructive ultrasonic investigation includes measuring the velocity of ultrasonic energy waves in lower order plate modes in one or more directions, and measuring phase velocity dispersion of higher order modes of the plate or sheet if needed. Texture or preferred grain orientation can be derived from these measurements with improved reliability and accuracy. The method can be utilized in production on moving metal plate or sheet. 9 figs.

A novel method for characterizing the impact response of functionally graded plates

NASA Astrophysics Data System (ADS)

Larson, Reid A.

Functionally graded material (FGM) plates are advanced composites with properties that vary continuously through the thickness of the plate. Metal-ceramic FGM plates have been proposed for use in thermal protection systems where a metal-rich interior surface of the plate gradually transitions to a ceramic-rich exterior surface of the plate. The ability of FGMs to resist impact loads must be demonstrated before using them in high-temperature environments in service. This dissertation presents a novel technique by which the impact response of FGM plates is characterized for low-velocity, low- to medium-energy impact loads. An experiment was designed where strain histories in FGM plates were collected during impact events. These strain histories were used to validate a finite element simulation of the test. A parameter estimation technique was developed to estimate local material properties in the anisotropic, non-homogenous FGM plates to optimize the finite element simulations. The optimized simulations captured the physics of the impact events. The method allows research & design engineers to make informed decisions necessary to implement FGM plates in aerospace platforms.

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.

Applications of a General Finite-Difference Method for Calculating Bending Deformations of Solid Plates

NASA Technical Reports Server (NTRS)

Walton, William C., Jr.

1960-01-01

This paper reports the findings of an investigation of a finite - difference method directly applicable to calculating static or simple harmonic flexures of solid plates and potentially useful in other problems of structural analysis. The method, which was proposed in doctoral thesis by John C. Houbolt, is based on linear theory and incorporates the principle of minimum potential energy. Full realization of its advantages requires use of high-speed computing equipment. After a review of Houbolt's method, results of some applications are presented and discussed. The applications consisted of calculations of the natural modes and frequencies of several uniform-thickness cantilever plates and, as a special case of interest, calculations of the modes and frequencies of the uniform free-free beam. Computed frequencies and nodal patterns for the first five or six modes of each plate are compared with existing experiments, and those for one plate are compared with another approximate theory. Beam computations are compared with exact theory. On the basis of the comparisons it is concluded that the method is accurate and general in predicting plate flexures, and additional applications are suggested. An appendix is devoted t o computing procedures which evolved in the progress of the applications and which facilitate use of the method in conjunction with high-speed computing equipment.

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.

The Nonlinear Dynamic Response of an Elastic-Plastic Thin Plate under Impulsive Loading,

DTIC Science & Technology

1987-06-11

Among those numerical methods, the finite element method is the most effective one. The method presented in this paper is an " influence function " numerical...computational time is much less than the finite element method. Its precision is higher also. II. Basic Assumption and the Influence Function of a Simple...calculation. Fig. 1 3 2. The Influence function of a Simple Supported Plate The motion differential equation of a thin plate can be written as DV’w+ _.eluq() (1

Bactericidal effects of a high-power, red light-emitting diode on two periodontopathic bacteria in antimicrobial photodynamic therapy in vitro.

PubMed

Umeda, Makoto; Tsuno, Akiko; Okagami, Yoshihide; Tsuchiya, Fumito; Izumi, Yuichi; Ishikawa, Isao

2011-11-01

  Light-emitting diodes have been investigated as new light activators for photodynamic therapy. We investigated the bactericidal effects of high-power, red light-emitting diodes on two periodontopathic bacteria in vitro.   A light-emitting diode (intensity: 1100 mW/cm(2) , peak wavelength: 650 nm) was used to irradiate a bacterial solution for either 10 or 20 s. Bacterial solutions (Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans) at a concentration of 2.5 × 10(6) c.f.u./mL were mixed with an equal volume of either methylene blue or toluidine blue O (0-20 μg/mL) and added to titer plate wells. The plate wells were irradiated with red light-emitting diode light from a distance of 22 or 40 mm. The contents were diluted, and 50 μL was smeared onto blood agar plates. After 1 week of culturing, bacterial c.f.u. were counted.   The light-emitting diode energy density was estimated to be approximately 4 and 8 J/cm(2) after 10 and 20 s of irradiation, respectively. Red light-emitting diode irradiation for 10 s from a distance of 22 mm, combined with methylene blue at concentrations >10 μg/mL, completely killed Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans.   High-power, red light-emitting diode irradiation with a low concentration of dye showed effective bactericidal effects against two periodontopathic bacteria. © 2011 Blackwell Publishing Asia Pty Ltd.

Comparison between open plating versus minimally invasive plate osteosynthesis for acute displaced clavicular shaft fractures.

PubMed

Sohn, Hoon-Sang; Kim, Won Ju; Shon, Min Soo

2015-08-01

Current literatures describe good clinical outcomes of acute displaced fracture of clavicle treated with minimally invasive plate osteosynthesis (MIPO). But, there are little comparative data of the outcomes between open plating and MIPO techniques. We compared the outcomes of open plating and MIPO for treatment of acute displaced clavicular shaft fractures. The author performed a retrospective review on a consecutive series of patients with clavicular shaft fracture who underwent open plating or MIPO. Fourteen patients were treated with open plating with interfragmentary screw fixation, and 19 were treated with the MIPO technique without exposing a fracture site itself. A superior plating method was applied to both groups. Patient demographics, clinical outcomes using Constant score and University of California Los Angeles (UCLA) shoulder score, operation time, union rate, complications, and radiographic evaluation were evaluated. There were no statistically significant differences in the demographic data, including patient's variables (age, gender, involved side, smoking, alcohol, and diabetic status) and fracture characteristics (trauma mechanism, distribution of fracture type, presence of polytrauma, and time from trauma to surgery) between the two groups. Mean operation time was 87.5 min in open plating and 77.2 min in MIPO (p=0.129). The mean time to union was 15.7 weeks in patients who underwent open plating and 16.8 weeks in patients who underwent MIPO (p=0.427). Although there was no significant difference, nonunion developed 1 case in MIPO while none was in open plating. Four patients in open plating had skin numbness (none in MIPO, p=0.024). There was no significant difference in the Constant score and UCLA score of the two surgical methods. This study showed that both open plating with interfragmentary screw fixation (Open plating) and minimally invasive plate osteosynthesis (MIPO) are equally effective and safe treatment methods for acute displaced clavicle shaft fracture. Copyright © 2015 Elsevier Ltd. All rights reserved.

Pollution impacts on bacterioplankton diversity in a tropical urban coastal lagoon system.

PubMed

Salloto, Gigliola R B; Cardoso, Alexander M; Coutinho, Felipe H; Pinto, Leonardo H; Vieira, Ricardo P; Chaia, Catia; Lima, Joyce L; Albano, Rodolpho M; Martins, Orlando B; Clementino, Maysa M

2012-01-01

Despite a great number of published studies addressing estuarine, freshwater and marine bacterial diversity, few have examined urban coastal lagoons in tropical habitats. There is an increasing interest in monitoring opportunistic pathogens as well as indigenous microbial community members in these water bodies by current molecular and microbiological approaches. In this work, bacterial isolates were obtained through selective plate dilution methods to evaluate antibiotic resistances. In addition, 16S rRNA gene libraries were prepared from environmental waters and mixed cultures grown in BHI medium inoculated with Jacarepaguá lagoon waters. Denaturing gradient gel electrophoresis (DGGE) analyses showed distinct community profiles between environmental communities from each studied site and their cultured counterparts. A total of 497 bacterial sequences were analyzed by MOTHUR, yielding 245 operational taxonomic units (OTUs) grouped at 97% similarity. CCA diagrams showcased how several environmental variables affect the distribution of 18 bacterial orders throughout the three distinct habitats. UniFrac metrics and Venn diagrams revealed that bacterial communities retrieved through each experimental approach were significantly different and that only one OTU, closely related to Vibrio cholerae, was shared between them. Potentially pathogenic bacteria were isolated from most sampled environments, fifty percent of which showed antibiotic resistance.

Pollution Impacts on Bacterioplankton Diversity in a Tropical Urban Coastal Lagoon System

PubMed Central

Salloto, Gigliola R. B.; Cardoso, Alexander M.; Coutinho, Felipe H.; Pinto, Leonardo H.; Vieira, Ricardo P.; Chaia, Catia; Lima, Joyce L.; Albano, Rodolpho M.; Martins, Orlando B.; Clementino, Maysa M.

2012-01-01

Despite a great number of published studies addressing estuarine, freshwater and marine bacterial diversity, few have examined urban coastal lagoons in tropical habitats. There is an increasing interest in monitoring opportunistic pathogens as well as indigenous microbial community members in these water bodies by current molecular and microbiological approaches. In this work, bacterial isolates were obtained through selective plate dilution methods to evaluate antibiotic resistances. In addition, 16S rRNA gene libraries were prepared from environmental waters and mixed cultures grown in BHI medium inoculated with Jacarepaguá lagoon waters. Denaturing gradient gel electrophoresis (DGGE) analyses showed distinct community profiles between environmental communities from each studied site and their cultured counterparts. A total of 497 bacterial sequences were analyzed by MOTHUR, yielding 245 operational taxonomic units (OTUs) grouped at 97% similarity. CCA diagrams showcased how several environmental variables affect the distribution of 18 bacterial orders throughout the three distinct habitats. UniFrac metrics and Venn diagrams revealed that bacterial communities retrieved through each experimental approach were significantly different and that only one OTU, closely related to Vibrio cholerae, was shared between them. Potentially pathogenic bacteria were isolated from most sampled environments, fifty percent of which showed antibiotic resistance. PMID:23226484

Assessing the antibiotic potential of essential oils against Haemophilus ducreyi.

PubMed

Lindeman, Zachary; Waggoner, Molly; Batdorff, Audra; Humphreys, Tricia L

2014-05-27

Haemophilus ducreyi is the bacterium responsible for the genital ulcer disease chancroid, a cofactor for the transmission of HIV, and it is resistant to many antibiotics. With the goal of exploring possible alternative treatments, we tested essential oils (EOs) for their efficacy as antimicrobial agents against H. ducreyi. We determine the minimum inhibitory concentration (MIC) of Cinnamomum verum (cinnamon), Eugenia caryophyllus (clove) and Thymus satureioides (thyme) oil against 9 strains of H. ducreyi using the agar dilution method. We also determined the minimum lethal concentration for each oil by subculturing from the MIC plates onto fresh agar without essential oil. For both tests, we used a 2-way ANOVA to evaluate whether antibiotic-resistant strains had a different sensitivity to the oils relative to non-resistant strains. All 3 oils demonstrated excellent activity against H. ducreyi, with MICs of 0.05 to 0.52 mg/mL and MLCs of 0.1-0.5 mg/mL. Antibiotic-resistant strains of H. ducreyi were equally susceptible to these 3 essential oils relative to non-resistant strains (p=0.409). E. caryophyllus, C. verum and T. satureioides oils are promising alternatives to antibiotic treatment for chancroid.

[USE OF LASER RAMAN-LUMINESCENT TECHNOLOGIES FOR EVALUATION OF QUALITY OF MEAT PRODUCTS AND DETERMINATION OF THE DEGREE OF THEIR BACTERIAL CONTAMINATION].

PubMed

Kukushkin, V I; Satusheva, E V; Aleksandrov, M T; Morozova, O A; Pashkov, E P; Ambartsumyan, O A; Amosova, V A

2015-01-01

Determination of the effect of microorganisms on spoilage of meat products during various temperature regimes of storage by integral indexes of luminescent lines in their spectra and development of an algorithm of microorganism indication by an express method using laser Raman-luminescent spectroscopy. Minced meat from beef and pork was used. Determination of quantity of mesophilic aerobic and opportunistic-anaerobic microorganisms was carried out by serial 10-fold dilutions with subsequent parallel seeding into Rida count total 24 plates and Petri dishes with 5% blood agar. Sample study was carried out in luminescent software-hardware complex Enspectr L405 (a variant of Enspectr M software-hardware complexes). Meat spoilage was established to be caused to a large degree by Pseudomonas genus (P. fluorescens, P. putida, P. fragi et al.) bacterial growth. Raman-luminescent spectra of bacteria that compose microflora, characterizing and accompanying beef and pork spoilage, were measured and recorded into a database. The results obtained will allow to use this technique in the future for both express-indication and differentiation of microorganisms and express-evaluation of quality of meat products at all stages of their manufacturing, storage, transport and realization.

Detection of Escherichia Coli Bacteria in Wastewater by using Graphene as a Sensing Material

NASA Astrophysics Data System (ADS)

Wibowo, K. M.; Sahdan, M. Z.; Ramli, N. I.; Muslihati, A.; Rosni, N.; Tsen, V. H.; Saim, H.; Ahmad, S. A.; Sari, Y.; Mansor, Z.

2018-04-01

Graphene is a family of carbon bonded in hexagonal honeycomb crystalline structure that has many superior properties. It was very suitable to be applied on sensor application due to the superior properties on electrical, physical, and optical. Furthermore, graphene also provide a large detection area since it has 2D structure. In this research, we develop graphene as a nanosensor for detection of Escherichia coli (E. coli) bacteria. The sample E. coli bacteria were cultured from domestic wastewater by using plate culture method and then isolated to get pure single colony. The serial dilution was performed to create different concentration of bacteria. Field emission scanning electron microscope and biochemical test were performed to ensure the sample genuinely target E. coli that defined by the physical size and optical properties. Raman spectroscopy measurements were also performed on the grapheme films, and it was found that the ratio of G peak and D peak intensity changing do to the presence of E. coli. The electrical properties of graphene shows the increasing number of the bacteria 4 to 273 cfu result in decreasing the resistance from 4.371 to 3.903 ohm gradually.

Contamination of common spices in Saudi Arabia markets with potential mycotoxin-producing fungi

PubMed Central

Hashem, Mohamed; Alamri, Saad

2010-01-01

Fifteen spices obtained from common markets were examined for their mould profile. A total of 520 fungal isolates, representing 57 species, were recovered and identified from dried and ground spice samples on three different media using standard dilution plate method. The most heavily contaminated spice samples examined were observed in ginger in order of magnitude of 5325–6800 cfu/g. The most predominant fungal genera encountered were Aspergillus, Penicillium, and Rhizopus. Relative occurrence values of taxa disclosed ranged between 80% for Aspergillusflavus, Aspergillusniger and Penicilliumarenicola, and 10% for some species. Samples obtained from sumac encountered very rare colony counts indicating its antifungal prosperities. The present magnitude of contamination and spectra of mycobiota approximate those reported for similar spice samples. Several potentially mycotoxigenic fungi were isolated from the majority of samples. The present study attracts the attention to potential risk for mycotoxins contamination may be caused as a result of using these spices, especially in great quantities. The study strongly recommends reduction in application of heavily contaminated spices like ginger in food processing and using some others like clove and sumac due to their antimicrobial properties. PMID:23961074

In Vitro Activity of p-Hydroxybenzyl Penicillin (Penicillin X) and Five Other Penicillins Against Neisseria gonorrhoeae: Comparisons of Strains from Patients with Uncomplicated Infections and from Women with Pelvic Inflammatory Disease

PubMed Central

Sackel, Stephen G.; Alpert, Susan; Rosner, Bernard; McCormack, William M.; Finland, Maxwell

1977-01-01

Minimum inhibitory concentrations (MICs) of six penicillins against 95 strains of Neisseria gonorrhoeae from patients with uncomplicated anogenital infections and 22 strains from women with pelvic inflammatory disease were determined by an agar plate dilution method, using an inocula replicator. Against all 117 strains, the order of activity observed was: BL-P1654 > penicillin X > penicillin G > ampicillin > amoxicillin = carbenicillin. MICs against strains isolated from women with gonococcal pelvic inflammatory disease were significantly higher than those against isolates from uncomplicated infections: BL-P1654, P < 0.001; penicillin X, P < 0.001; penicillin G, P < 0.001; ampicillin, P < 0.001; and amoxicillin, P < 0.05. MICs of penicillin G were ≥0.125 μg/ml against 33 (36%) of the 92 strains from patients with uncomplicated infections, as contrasted with 15 (68%) of the 22 isolates from women with pelvic inflammatory disease (P < 0.01). The means of the MICs of penicillin G were 0.06 μg/ml for the former and 0.14 μg/ml for the latter. PMID:407840

Replication of Syngrapha falcifera Multiple-Nuclear Polyhedrosis Virus-D in Different Insect Cells

NASA Astrophysics Data System (ADS)

Khalid Nessr Alhag, Sadeq; Xin, Peng Jian

Six insect cell lines were tested for susceptibility to Syngrapha falcifera multiple nucleocapsid nucleopolyhedrovirus-D (SfaMNPV-D) infection by use of a typical endpoint assay procedure. Cell lines from Trichoplusia ni (Tn5B1-4), (L105-clone), Spodoptera litura (SL-ZSU-1), Spodoptera frugiperda (IPLB-SF-21), Pieris rapaeb (Pr-E-HNU9) and Helicoverpa zea (BCIRL-HZ-AM1) in 96-well tissue culture plates were infected with dilutions of extra cellular virus suspensions of (SfaMNPV-D). Each cell/virus combination was incubated at temperatures 27°C and wells were scored for positive infection at 2 to 4 day intervals. The resulting data were analyzed by Reed and Muench method, providing virus titers for each combination of virus, cell line. The results were categorized by accuracy and by rapidity of maximum titer. Virus titer of Tn5B-4 was higher than other cell lines TCID50 8.7x108, the lowest level detected in infected was in (Pr-E-HNU9) cells TCID50 2.4x108. No Virions or polyhedral inclusion bodies were detected in infected SL-ZSU-1 cells.

A correlation method to predict the surface pressure distribution of an infinite plate or a body of revolution from which a jet is issuing

NASA Technical Reports Server (NTRS)

Perkins, S. C., Jr.; Mendenhall, M. R.

1980-01-01

A correlation method to predict pressures induced on an infinite plate by a jet exhausting normal to the plate into a subsonic free stream was extended to jets exhausting at angles to the plate and to jets exhausting normal to the surface of a body revolution. The complete method consisted of an analytical method which models the blockage and entrainment properties of the jet and an empirical correlation which accounts for viscous effects. For the flat plate case, the method was applicable to jet velocity ratios up to ten, jet inclination angles up to 45 deg from the normal, and radial distances up to five diameters from the jet. For the body of revolution case, the method was applicable to a body at zero degrees angle of attack, jet velocity ratios 1.96 and 3.43, circumferential angles around the body up to 25 deg from the jet, axial distances up to seven diameters from the jet, and jet-to-body diameter ratios less than 0.1.

Identification of damage in plates using full-field measurement with a continuously scanning laser Doppler vibrometer system

NASA Astrophysics Data System (ADS)

Chen, Da-Ming; Xu, Y. F.; Zhu, W. D.

2018-05-01

An effective and reliable damage identification method for plates with a continuously scanning laser Doppler vibrometer (CSLDV) system is proposed. A new constant-speed scan algorithm is proposed to create a two-dimensional (2D) scan trajectory and automatically scan a whole plate surface. Full-field measurement of the plate can be achieved by applying the algorithm to the CSLDV system. Based on the new scan algorithm, the demodulation method is extended from one dimension for beams to two dimensions for plates to obtain a full-field operating deflection shape (ODS) of the plate from velocity response measured by the CSLDV system. The full-field ODS of an associated undamaged plate is obtained by using polynomials with proper orders to fit the corresponding full-field ODS from the demodulation method. A curvature damage index (CDI) using differences between curvatures of ODSs (CODSs) associated with ODSs that are obtained by the demodulation method and the polynomial fit is proposed to identify damage. An auxiliary CDI obtained by averaging CDIs at different excitation frequencies is defined to further assist damage identification. An experiment of an aluminum plate with damage in the form of 10.5% thickness reduction in a damage area of 0.86% of the whole scan area is conducted to investigate the proposed method. Six frequencies close to natural frequencies of the plate and one randomly selected frequency are used as sinusoidal excitation frequencies. Two 2D scan trajectories, i.e., a horizontally moving 2D scan trajectory and a vertically moving 2D scan trajectory, are used to obtain ODSs, CODSs, and CDIs of the plate. The damage is successfully identified near areas with consistently high values of CDIs at different excitation frequencies along the two 2D scan trajectories; the damage area is also identified by auxiliary CDIs.

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

Plate measurement techniques and reduction methods used by the West German satellite observers, and resulting consequences for the observation

NASA Technical Reports Server (NTRS)

Deker, H.

1971-01-01

The West German tracking stations are equipped with ballistic cameras. Plate measurement and plate reduction must therefore follow photogrammetric methods. Approximately 100 star positions and 200 satellite positions are measured on each plate. The mathematical model for spatial rotation of the bundle of rays is extended by including terms for distortion and internal orientation of the camera as well as by providing terms for refraction which are computed for the measured coordinates of the star positions on the plate. From the measuring accuracy of the plate coordinates it follows that the timing accuracy for the exposures has to be about one millisecond, in order to obtain a homogeneous system.

[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.

Effusion plate using additive manufacturing methods

DOEpatents

Johnson, Thomas Edward; Keener, Christopher Paul; Ostebee, Heath Michael; Wegerif, Daniel Gerritt

2016-04-12

Additive manufacturing techniques may be utilized to construct effusion plates. Such additive manufacturing techniques may include defining a configuration for an effusion plate having one or more internal cooling channels. The manufacturing techniques may further include depositing a powder into a chamber, applying an energy source to the deposited powder, and consolidating the powder into a cross-sectional shape corresponding to the defined configuration. Such methods may be implemented to construct an effusion plate having one or more channels with a curved cross-sectional geometry.

Method of obtaining intensified image from developed photographic films and plates

NASA Technical Reports Server (NTRS)

Askins, B. S. (Inventor)

1978-01-01

A method is explained of obtaining intensified images from silver images on developed photographic films and plates. The steps involve converting silver of the developed film or plate to a radioactive compound by treatment with an aqueous alkaline solution of an organo-S35 compound; placing the treated film or plate in direct contact with a receiver film which is then exposed by radiation from the activated film; and developing and fixing the resulting intensified image on the receiver film.

Reliability assessment of different plate theories for elastic wave propagation analysis in functionally graded plates.

PubMed

Mehrkash, Milad; Azhari, Mojtaba; Mirdamadi, Hamid Reza

2014-01-01

The importance of elastic wave propagation problem in plates arises from the application of ultrasonic elastic waves in non-destructive evaluation of plate-like structures. However, precise study and analysis of acoustic guided waves especially in non-homogeneous waveguides such as functionally graded plates are so complicated that exact elastodynamic methods are rarely employed in practical applications. Thus, the simple approximate plate theories have attracted much interest for the calculation of wave fields in FGM plates. Therefore, in the current research, the classical plate theory (CPT), first-order shear deformation theory (FSDT) and third-order shear deformation theory (TSDT) are used to obtain the transient responses of flexural waves in FGM plates subjected to transverse impulsive loadings. Moreover, comparing the results with those based on a well recognized hybrid numerical method (HNM), we examine the accuracy of the plate theories for several plates of various thicknesses under excitations of different frequencies. The material properties of the plate are assumed to vary across the plate thickness according to a simple power-law distribution in terms of volume fractions of constituents. In all analyses, spatial Fourier transform together with modal analysis are applied to compute displacement responses of the plates. A comparison of the results demonstrates the reliability ranges of the approximate plate theories for elastic wave propagation analysis in FGM plates. Furthermore, based on various examples, it is shown that whenever the plate theories are used within the appropriate ranges of plate thickness and frequency content, solution process in wave number-time domain based on modal analysis approach is not only sufficient but also efficient for finding the transient waveforms in FGM plates. Copyright © 2013 Elsevier B.V. All rights reserved.

The Development and Evaluation of a Portion Plate for Youth: A Pilot Study

ERIC Educational Resources Information Center

Bohnert, Amy M.; Randall, Edin T.; Tharp, Stephanie; Germann, Julie

2011-01-01

Objective: To develop and evaluate a portion plate for adolescents (Nutri-plate). Methods: Sixteen African American adolescents (mean age = 12.94 years; 66% male) were randomized to participate in either plate design or nutrition education sessions. Adolescents' input was used to create the Nutri-plate, and participants' food selection and intake…

Detection and estimation of defects in a circular plate using operational deflection shapes

NASA Astrophysics Data System (ADS)

Pai, Perngjin F.; Oh, Yunje; Kim, Byeong-Seok

2002-06-01

This paper investigates dynamic characteristics (mode shapes and natural frequencies) and defect detection of circular plates using a scanning laser vibrometer. Exact dynamic characteristics of a circular aluminum plate having a clamped inner rim and a free outer rim are obtained using two methods; one uses Bessel functions and the other uses a multiple shooting method. An in-house finite element code GESA is also used to analyze the circular plate using the DKT plate element. Numerical results show that some reports in the literature are incorrect and that high-frequency Operational Deflection Shapes (ODSs) are needed in order to locate small defects. Detection of two defects in the circular aluminum plate is experimentally studied using the distributions of RMS velocities under broadband periodic chirp excitations. RMS velocities of ODSs, symmetry breaking of ODSs, splitting of natural frequencies and ODSs, and a Boundary Effect Detection (BED) method. The BED method is non-destructive and model-independent; it processes experimental ODSs to reveal extra local boundary effects caused by defects to reveal locations of defects. Experimental results show that small defects in circular plates can be pinpointed by these approaches. Moreover, a new concept of using the balance of elastic and kinetic energies within a mode cell for detecting defects in two- dimensional structures of irregular shapes is proposed.

Enhancements for digital imaging of gusset plate connections : fisheye and image stitching.

DOT National Transportation Integrated Search

2011-09-01

This report describes techniques to convert fisheye digital images of gusset plates to perspective images (defish). It also describes methods to stitch together partial images of a gusset plate into a composite of the complete gusset plate. The conve...

Comparative Study of Hydroalcoholic Extracts of Momordica charantia L. against Foodborne Pathogens

PubMed Central

Rakholiya, Kalpna; Vaghela, P.; Rathod, T.; Chanda, Sumitra

2014-01-01

The antimicrobial effect of 24 different hydroalcoholic extracts (100, 75, 50 and 25% methanol and water) obtained from four parts (leaf+stem (aerial), peel, pulp and seed) of Momordica charantia L. were investigated against five Gram-positive, six Gram-negative and four fungal strains. The extraction was done by individual cold percolation method using hexane, different hydroalcoholic solvent (100, 75, 50 and 25% methanol) and water. The antimicrobial activity was done by agar well diffusion assay. The extracts, which showed >15 mm zone of inhibition, were further screened to determine minimum inhibitory concentration and minimum bactericidal concentration using a broth dilution method performed in 96-well microtitre plate. The extractive yield was highest in aqueous extracts of all the four parts closely followed by 25% methanol. Micrococcus flavus was the most susceptible Gram-positive bacteria and Pseudomonas testosteroni was the most susceptible Gram-negative bacteria. The highest antibacterial activity was shown by 100% methanol. The Gram-negative Pseudomonas spp. was more susceptible towards all the extracts than the Gram-positive bacteria or fungal strains investigated. One hundred percent and 50% methanol extracts of seed showed lowest minimum inhibitory concentration and minimum bactericidal concentration values, that is <39 and 625 μg/ml, respectively, against Pseudomonas pictorum. Therefore, these extracts would be of interest in the control of Pseudomonas spp. in food industry as well as used for therapeutic purposes. PMID:24843188

Treatment of Palm Oil Mill Effluent by a Microbial Consortium Developed from Compost Soils

PubMed Central

Nwuche, Charles O.; Ogbonna, James C.

2014-01-01

A method for the aerobic treatment of palm oil mill effluent (POME) was investigated in shake-flask experiments using a consortium developed from POME compost. POME was initially centrifuged at 4,000 g for 15 min and the supernatant was enriched with (NH4)2SO4 (0.5%) and yeast extract (0.25%) to boost its nitrogen content. At optimum pH (pH 4) and temperature (40°C) conditions, the chemical oxygen demand (COD) of the effluent decreased from 10,350 to 1,000 mg/L (90.3%) after 7 days. The total bacterial population determined by plate count enumeration was 2.4 × 106 CFU/mL, while the fungal count was 1.8 × 103 colonies/mL. Bacteria of the genera Pseudomonas, Flavobacterium, Micrococcus, and Bacillus were isolated, while the fungal genera included Aspergillus, Penicillium, Trichoderma, and Mucor. When the isolated species were each inoculated into separate batches of the raw effluent, both pH and COD were unchanged. However, at 75 and 50% POME dilutions, the COD dropped by 52 and 44%, respectively, while the pH increased from 4 to 7.53. POME treatment by aerobic method is sustainable and holds promising prospects for cushioning the environment from the problems associated with the use of anaerobic systems. PMID:27433536

Action of Monomeric/Gemini Surfactants on Free Cells and Biofilm of Asaia lannensis.

PubMed

Koziróg, Anna; Kręgiel, Dorota; Brycki, Bogumił

2017-11-22

We investigated the biological activity of surfactants based on quaternary ammonium compounds: gemini surfactant hexamethylene-1,6-bis-( N,N -dimethyl- N -dodecylammonium bromide) (C6), synthesized by the reaction of N,N -dimethyl- N- dodecylamine with 1,6-dibromohexane, and its monomeric analogue dodecyltrimethylammonium bromide (DTAB). The experiments were performed with bacteria Asaia lannensis , a common spoilage in the beverage industry. The minimal inhibitory concentration (MIC) values were determined using the tube standard two-fold dilution method. The growth and adhesive properties of bacterial cells were studied in different culture media, and the cell viability was evaluated using plate count method. Both of the surfactants were effective against the bacterial strain, but the MIC of gemini compound was significantly lower. Both C6 and DTAB exhibited anti-adhesive abilities. Treatment with surfactants at or below MIC value decreased the number of bacterial cells that were able to form biofilm, however, the gemini surfactant was more effective. The used surfactants were also found to be able to eradicate mature biofilms. After 4 h of treatment with C6 surfactant at concentration 10 MIC, the number of bacterial cells was reduced by 91.8%. The results of this study suggest that the antibacterial activity of the gemini compound could make it an effective microbiocide against the spoilage bacteria Asaia sp. in both planktonic and biofilm stages.

Efficient High-Fidelity, Geometrically Exact, Multiphysics Structural Models

DTIC Science & Technology

2011-10-14

fuctionally graded core. International Journal for Numerical Methods in Engineering, 68:940– 966, 2006. 7F. Shang, Z. Wang, and Z. Li. Analysis of...normal deformable plate theory and MLPG method with radial basis fuctions . Composite Structures, 80:539– 552, 2007. 17W. Zhen and W. Chen. A higher-order...functionally graded plates by using higher-order shear and normal deformable plate theory and MLPG method with radial basis fuctions . Composite Structures, 80

Method for plating with metal oxides

DOEpatents

Silver, Gary L.; Martin, Frank S.

1994-08-23

A method of plating hydrous metal oxides on at least one substrate, which method is indifferent to the electrochemical properties of the substrate, and comprises reacting metallic ions in aqueous solution with an appropriate oxidizing agent such as sodium hypochlorite or calcium sulfite with oxygen under suitable conditions of pH and concentration such that oxidation and precipitation of metal oxide are sufficiently slow to allow satisfactory plating of metal oxide on the substrate.

Wave propagation in a multilayered laminated cross-ply composite plate

NASA Technical Reports Server (NTRS)

Shah, A. H.; Datta, S. K.; Karunasena, W.

1991-01-01

Dispersion of guided waves in a cross-ply laminated plate has been studied here using a stiffness method and an exact method. It is shown that the number of laminae strongly influences the dispersion behavior. Further, it is found that when the number of laminae is sufficiently large, then the dispersion behavior can be predicted by treating the plate as homogeneous with six stiffness constants obtained by using an effective modulus method.