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Sample records for 96-well culture plates

  1. Quantification of cells cultured on 96-well plates.

    PubMed

    Kueng, W; Silber, E; Eppenberger, U

    1989-10-01

    The method for cell number measurement in monolayer cultures by crystal violet staining published recently by Gillies et al. (R. G. Gillies, N. Didier, M. Denton (1986) Anal. Biochem. 159, 109-113) was modified and significantly improved. The procedure was adapted for use in 96-well plates since the method is inherently very sensitive. Modifications allowed fast and complete solubilization of dye adsorbed by cell nuclei during staining. Since light absorption of the unstained or destained cell layers is negligible, cell number measurements can be performed in the respective wells. Due to these features, multiple assays may be carried out rapidly using standard 96-well plate readers. In addition, it is shown that the sensitivity of the assay can be varied and easily controlled by choosing the appropriate pH during the staining procedure. This increases the flexibility of the method making it useful for determining cell density of a wide range of different cell types. PMID:2604040

  2. Toward Microbioreactor Arrays: A Slow-Responding Oxygen Sensor for Monitoring of Microbial Cultures in Standard 96-Well Plates.

    PubMed

    Glauche, Florian; John, Gernot T; Arain, Sarina; Knepper, Andreas; Neubauer, Antje; Goelling, Detlef; Lang, Christine; Violet, Norman; King, Rudibert; Neubauer, Peter

    2015-08-01

    In this study, a slow-responding chemo-optical sensor for dissolved oxygen (DO) integrated into a 96-well plate was developed. The slow response time ensures that the measured oxygen value does not change much during plate transport to the microplate reader. The sensor therefore permits at-line DO measurement of microbial cultures. Moreover, it eliminates the necessity of individual optical measurement systems for each culture plate, as many plates can be measured successively. Combined with the 96-well format, this increases the experimental throughput enormously. The novel sensor plate (Slow OxoPlate) consists of fluorophores suspended in a polymer matrix that were placed into u-bottom 96-well plates. Response time was measured using sodium sulfite, and a t90 value of 9.7 min was recorded. For application, DO values were then measured in Escherichia coli and Saccharomyces cerevisiae cultures grown under fed-batch-like conditions. Depending on the DO sensor's response time, different information on the oxygenation state of the culture plate was obtained: a fast sensor variant detects disturbance through sampling, whereas the slow sensor indicates oxygen limitation during incubation. A combination of the commercially available OxoPlate and the Slow OxoPlate enables operators of screening facilities to validate their cultivation procedures with regard to oxygen availability. PMID:25720599

  3. Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System

    PubMed Central

    Conway, Michael K.; Gerger, Michael J.; Balay, Erin E.; O'Connell, Rachel; Hanson, Seth; Daily, Neil J.; Wakatsuki, Tetsuro

    2015-01-01

    Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells in regenerative medicine, drug discovery and safety testing. In order to produce stem cell derived biopharmaceutics and cells for tissue engineering and transplantation, a cost-effective cell-manufacturing technology is essential. Maintenance of pluripotency and stable performance of cells in downstream applications (e.g., cell differentiation) over time is paramount to large scale cell production. Yet that can be difficult to achieve especially if cells are cultured manually where the operator can introduce significant variability as well as be prohibitively expensive to scale-up. To enable high-throughput, large-scale stem cell production and remove operator influence novel stem cell culture protocols using a bench-top multi-channel liquid handling robot were developed that require minimal technician involvement or experience. With these protocols human induced pluripotent stem cells (iPSCs) were cultured in feeder-free conditions directly from a frozen stock and maintained in 96-well plates. Depending on cell line and desired scale-up rate, the operator can easily determine when to passage based on a series of images showing the optimal colony densities for splitting. Then the necessary reagents are prepared to perform a colony split to new plates without a centrifugation step. After 20 passages (~3 months), two iPSC lines maintained stable karyotypes, expressed stem cell markers, and differentiated into cardiomyocytes with high efficiency. The system can perform subsequent high-throughput screening of new differentiation protocols or genetic manipulation designed for 96-well plates. This technology will reduce the labor and technical burden to produce large numbers of identical stem cells for a myriad of applications. PMID:26068617

  4. Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System.

    PubMed

    Conway, Michael K; Gerger, Michael J; Balay, Erin E; O'Connell, Rachel; Hanson, Seth; Daily, Neil J; Wakatsuki, Tetsuro

    2015-01-01

    Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells in regenerative medicine, drug discovery and safety testing. In order to produce stem cell derived biopharmaceutics and cells for tissue engineering and transplantation, a cost-effective cell-manufacturing technology is essential. Maintenance of pluripotency and stable performance of cells in downstream applications (e.g., cell differentiation) over time is paramount to large scale cell production. Yet that can be difficult to achieve especially if cells are cultured manually where the operator can introduce significant variability as well as be prohibitively expensive to scale-up. To enable high-throughput, large-scale stem cell production and remove operator influence novel stem cell culture protocols using a bench-top multi-channel liquid handling robot were developed that require minimal technician involvement or experience. With these protocols human induced pluripotent stem cells (iPSCs) were cultured in feeder-free conditions directly from a frozen stock and maintained in 96-well plates. Depending on cell line and desired scale-up rate, the operator can easily determine when to passage based on a series of images showing the optimal colony densities for splitting. Then the necessary reagents are prepared to perform a colony split to new plates without a centrifugation step. After 20 passages (~3 months), two iPSC lines maintained stable karyotypes, expressed stem cell markers, and differentiated into cardiomyocytes with high efficiency. The system can perform subsequent high-throughput screening of new differentiation protocols or genetic manipulation designed for 96-well plates. This technology will reduce the labor and technical burden to produce large numbers of identical stem cells for a myriad of applications. PMID:26068617

  5. Electrothermal micromixing in 96 well plate

    NASA Astrophysics Data System (ADS)

    Kauffmann, Paul; Loire, Sophie; Mezic, Igor

    2011-11-01

    Diagnostic and pharmacology processes could be greatly accelerated by appropriate mixing. Here electrothermal flows are explored to provide mixing of conductive physiological solutions (=1.6 S/m) in a 96 well plate. Three interdigitated electrodes provide an electric field (< 15Vpp, 1MHz) beneath each well. Polarization and conduction phenomenon of the fluid in a well will be first modeled numerically and compared to an electrical circuit model. Due to high conductivity and permittivity of the fluid, the impedance of the array of filled wells collapse dramatically (96 wells: R = 1Ohm, C=250nF). The power supply challenges accordingly raised by arrays of electrothermal micromixers will be then analyzed. The efficiency of different methods of mixing in those wells will be also compared: the addition of low frequency signal leading to AC electro-osmotic perturbations, a blinking vortices method. The experimental results will be compared to simulations.

  6. Lactobacillus casei microbiological assay of folic acid derivatives in 96-well microtiter plates.

    PubMed

    Horne, D W; Patterson, D

    1988-11-01

    Microbiological assay is still widely used for estimating folic acid derivatives in serum and other biological samples. We describe here a modification of this procedure involving use of 96-well microtiter plates. This procedure, used with modern, computer-interfaced microtiter-plate readers and data-reduction software, greatly shortens the time and minimizes reagent costs for this assay. Under the conditions of our assay procedures, all folic acid derivatives tested gave equal growth response for Lactobacillus casei. Results for assays of rat liver extracts showed excellent agreement between the standard bioassay and the 96-well procedure. PMID:3141087

  7. HIGHLY SENSITIVE ASSAY FOR ANTICHOLINESTERASE COMPOUNDS USING 96 WELL PLATE FORMAT

    EPA Science Inventory

    The rapid and sensitive detection of organophosphate insecticides using a 96 well plate format is reported. Several features of this assay make it attractive for development as a laboratory-based or field screening assay. Acetylcholinesterase (AChE) was stabilized in a gelati...

  8. A microassay for measuring glycogen in 96-well-cultured cells.

    PubMed

    Gómez-Lechón, M J; Ponsoda, X; Castell, J V

    1996-05-01

    This study describes a rapid, sensitive, and automated spectrophotometric enzymatic microassay that measures the intracellular glycogen of primary cultured hepatocytes and other cultured cells in 96-well plates and can be adapted for other samples that are transferred to these plates. The procedure involves in situ disruption of cells, followed by hydrolysis of glycogen into glucosyl units by fungal glucoamylase (exo-1.4-alpha-glucosidase, EC 3.2.1.3), and glucose determination with the glucose oxidase colorimetric method. The color intensity can be measured in conventional ELISA readers, and the data can be fed to an on-line computer for rapid processing. The advantages of this method are its simplicity and automation, the reduction in sample handling, and the small number of cells required compared to other conventional methods. PMID:8660508

  9. Biofilm Localization in the Vertical Wall of Shaking 96-Well Plates

    PubMed Central

    Gomes, Luciana C.; Moreira, Joana M. R.; Simões, Manuel; Melo, Luís F.; Mergulhão, Filipe J.

    2014-01-01

    Microtiter plates with 96 wells are being increasingly used for biofilm studies due to their high throughput, low cost, easy handling, and easy application of several analytical methods to evaluate different biofilm parameters. These methods provide bulk information about the biofilm formed in each well but lack in detail, namely, regarding the spatial location of the biofilms. This location can be obtained by microscopy observation using optical and electron microscopes, but these techniques have lower throughput and higher cost and are subjected to equipment availability. This work describes a differential crystal violet (CV) staining method that enabled the determination of the spatial location of Escherichia coli biofilms formed in the vertical wall of shaking 96-well plates. It was shown that the biofilms were unevenly distributed on the wall with denser cell accumulation near the air-liquid interface. The results were corroborated by scanning electron microscopy and a correlation was found between biofilm accumulation and the wall shear strain rates determined by computational fluid dynamics. The developed method is quicker and less expensive and has a higher throughput than the existing methods available for spatial location of biofilms in microtiter plates. PMID:24834360

  10. Stability test for amorphous materials in humidity controlled 96-well plates by near-infrared spectroscopy.

    PubMed

    Otsuka, Makoto; Tanabe, Hideaki

    2012-03-01

    The purpose of this research is to apply near infrared spectrometry (NIR) with chemoinformetrics to predict the change of crystalline properties of indomethacin (IMC) amorphous under various levels of relative humidity storage conditions. Stability test for amorphous and meta-stable polymorphic forms was performed in humidity controlled the modified 96-well quartz plates containing various kinds of saturated salt solutions (0-100% of relative humidity (RH)) by NIR spectroscopy. Amorphous form was obtained melt product to pour into liquid nitrogen and after then ground. Samples were stored at 25°C in the 6-well plates at various levels of RH. The spectra of the powder samples were measured by the reflectance FT-NIR spectrometer. The second derivative spectra of form α showed specific absorption peaks at 4980, 6036, 7296 and 8616 cm-1 and that of form γ showed those at 5020, 5028, 7344, 7428 and 8436 cm-1. After storage at less than 50% RH, the peak intensities at 5020, 5028, 7344, 7428 and 8436 cm-1 of the amorphous solid increased with increasing of storage time. However, the peak intensity at 4980, 6036 and 7296 cm-1 increased at more than 50% RH Please check and confirm the edit. The results suggested that at lower humidity, the IMC amorphous solid transformed into form γ, but it transformed into form α at more than high humidity. It is possible that crystalline stability of the pharmaceutical preparations could be predicted by using humidity controlled 96-well plates and reflectance NIR-chemoinformetric methods. PMID:21942281

  11. Measuring oxidative stress resistance of Caenorhabditis elegans in 96-well microtiter plates.

    PubMed

    Possik, Elite; Pause, Arnim

    2015-01-01

    Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic human disorders, including cardiovascular and neurodegenerative diseases, diabetes, aging, and cancer. Therefore, it is important to study oxidative stress not only in cell systems but also using whole organisms. C. elegans is an attractive model organism to study the genetics of oxidative stress signal transduction pathways, which are highly evolutionarily conserved. Here, we provide a protocol to measure oxidative stress resistance in C. elegans in liquid. Briefly, ROS-inducing reagents such as paraquat (PQ) and H2O2 are dissolved in M9 buffer, and solutions are aliquoted in the wells of a 96 well microtiter plate. Synchronized L4/young adult C. elegans animals are transferred to the wells (5-8 animals/well) and survival is measured every hour until most worms are dead. When performing an oxidative stress resistance assay using a low concentration of stressors in plates, aging might influence the behavior of animals upon oxidative stress, which could lead to an incorrect interpretation of the data. However, in the assay described herein, this problem is unlikely to occur since only L4/young adult animals are being used. Moreover, this protocol is inexpensive and results are obtained in one day, which renders this technique attractive for genetic screens. Overall, this will help to understand oxidative stress signal transduction pathways, which could be translated into better characterization of oxidative stress-associated human disorders. PMID:25993260

  12. A rapid direct telomerase assay method using 96-well streptavidin plates.

    PubMed

    Francis, Rawle; Friedman, Simon H

    2002-05-01

    We have developed a high-throughput direct assay methodfor the assay of telomerase activity that improves on previous direct telomerase assays in two ways that allow larger numbers of samples to be conveniently processed: (i) 96-well streptavidin coated plates are used to bind and wash biotinylated primer extension products from the telomerase assay, as opposed to tubes containing streptavidin-coated magnetic beads; and (ii) storage phosphor-imagery is used instead of film autoradiography to detect telomerase products after being washed and released from the streptavidin-derivatized matrix. This method improves on previous direct assay methods using magnetic beads by allowing larger numbers of samples to be conveniently assayed. Also, the total activity of the radiolabeled nucleotides used in this procedure is significantly lower than that used in standard direct telomerase assays, lowering costs and exposure to radioactivity. We have validated the assay by repeating, in triplicate, the IC50 determination of rivanol, our previously identified telomerase inhibitor. PMID:12019789

  13. Establishment of ATP-based luciferase viability assay in 96-well plate for Trypanosoma congolense.

    PubMed

    Suganuma, Keisuke; Allamanda, Puttik; Hakimi, Hassan; Zhou, Mo; Angeles, Jose Ma; Kawazu, Shin-ichiro; Inoue, Noboru

    2014-11-01

    Animal African trypanosomosis (AAT), caused by Trypanosoma congolense, is widespread throughout sub-Saharan Africa. There are significant concerns related to the current drugs available for the treatment of AAT due to their limited effectiveness across species and their adverse effects. Moreover, drug resistant trypanosomes have recently been reported in the field. High throughput screening (HTS) of large chemical compound library collections is a promising approach for identifying novel drug candidates. While HTS for Trypanozoon trypanosomes, T. brucei sspp. and T. evansi is well established, no assays have been developed for T. congolense. In the present study, the authors developed an ATP-based luciferase viability assay for T. congolense in a 96-well plate format. The calculated 50% inhibitory concentration (IC50) values for pentamidine and diminazene were 10-100 times higher in T. congolense than in T. brucei. This result suggests that the transporters for the 2 tested compounds differ between T. congolense and T. brucei. This assay could further be applied to screen novel chemical compounds for the treatment of AAT caused by T. congolense. PMID:25056575

  14. Establishment of ATP-Based Luciferase Viability Assay in 96-Well Plate for Trypanosoma congolense

    PubMed Central

    SUGANUMA, Keisuke; ALLAMANDA, Puttik; HAKIMI, Hassan; ZHOU, Mo; ANGELES, Jose Ma.; KAWAZU, Shin-ichiro; INOUE, Noboru

    2014-01-01

    ABSTRACT Animal African trypanosomosis (AAT), caused by Trypanosoma congolense, is widespread throughout sub-Saharan Africa. There are significant concerns related to the current drugs available for the treatment of AAT due to their limited effectiveness across species and their adverse effects. Moreover, drug resistant trypanosomes have recently been reported in the field. High throughput screening (HTS) of large chemical compound library collections is a promising approach for identifying novel drug candidates. While HTS for Trypanozoon trypanosomes, T. brucei sspp. and T. evansi is well established, no assays have been developed for T. congolense. In the present study, the authors developed an ATP-based luciferase viability assay for T. congolense in a 96-well plate format. The calculated 50% inhibitory concentration (IC50) values for pentamidine and diminazene were 10–100 times higher in T. congolense than in T. brucei. This result suggests that the transporters for the 2 tested compounds differ between T. congolense and T. brucei. This assay could further be applied to screen novel chemical compounds for the treatment of AAT caused by T. congolense. PMID:25056575

  15. Urine sample preparation in 96-well filter plates for quantitative clinical proteomics.

    PubMed

    Yu, Yanbao; Suh, Moo-Jin; Sikorski, Patricia; Kwon, Keehwan; Nelson, Karen E; Pieper, Rembert

    2014-06-01

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ~10 μg of total protein were processed by 96FASP and LC-MS resulting in 700-900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics. PMID:24797144

  16. IDENTIFICATION OF SALMONELLA-POSITIVE FECAL SAMPLES USING A 96-WELL MICROCULTURE PLATE TECHNIQUE (RX METHOD)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conventional Salmonella isolation involves multiple sample transfers to culture media performed by an experienced microbiologist. The modified semi-solid RV and XLT (RX) Plate method, a modification of the RX tube format designed by Gailey et al. (2004), consolidates pre-enrichment (buffered pepton...

  17. SIMPLE DEVICE TO DELIVER BEADS TO 96-WELL PLATES FOR RAPID RESUSPENSION OF CELLS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genomic sequencing involves sequence determination of inserts from a large numbers of plasmids of a genomic library. In high throughput DNA sequencing projects, cultures are grown in a 96-deep well plate format, centrifuged, and resuspended. Resuspension of cells was aided by use of a simple devic...

  18. Ice-Cap: a method for growing Arabidopsis and tomato plants in 96-well plates for high-throughput genotyping.

    PubMed

    Su, Shih-Heng; Clark, Katie A; Gibbs, Nicole M; Bush, Susan M; Krysan, Patrick J

    2011-01-01

    It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping project is to collect a tissue sample from each individual plant. The traditional approach to this task is to sample plants one-at-a-time. If one wishes to genotype hundreds or thousands of individuals, however, using this strategy results in a significant bottleneck in the genotyping pipeline. The Ice-Cap method that we describe here provides a high-throughput solution to this challenge by allowing one scientist to collect tissue from several thousand seedlings in a single day (1,2). This level of throughput is made possible by the fact that tissue is harvested from plants 96-at-a-time, rather than one-at-a-time. The Ice-Cap method provides an integrated platform for performing seedling growth, tissue harvest, and DNA extraction. The basis for Ice-Cap is the growth of seedlings in a stacked pair of 96-well plates. The wells of the upper plate contain plugs of agar growth media on which individual seedlings germinate. The roots grow down through the agar media, exit the upper plate through a hole, and pass into a lower plate containing water. To harvest tissue for DNA extraction, the water in the lower plate containing root tissue is rapidly frozen while the seedlings in the upper plate remain at room temperature. The upper plate is then peeled away from the lower plate, yielding one plate with 96 root tissue samples frozen in ice and one plate with 96 viable seedlings. The technique is named "Ice-Cap" because it uses ice to capture the root tissue. The 96-well plate containing the seedlings can then wrapped in foil and transferred to low temperature. This process suspends further growth of the seedlings, but does not affect their viability. Once genotype analysis has been completed, seedlings with the desired genotype can be transferred from the 96-well plate to soil for further propagation. We have demonstrated

  19. MStern Blotting-High Throughput Polyvinylidene Fluoride (PVDF) Membrane-Based Proteomic Sample Preparation for 96-Well Plates.

    PubMed

    Berger, Sebastian T; Ahmed, Saima; Muntel, Jan; Cuevas Polo, Nerea; Bachur, Richard; Kentsis, Alex; Steen, Judith; Steen, Hanno

    2015-10-01

    We describe a 96-well plate compatible membrane-based proteomic sample processing method, which enables the complete processing of 96 samples (or multiples thereof) within a single workday. This method uses a large-pore hydrophobic PVDF membrane that efficiently adsorbs proteins, resulting in fast liquid transfer through the membrane and significantly reduced sample processing times. Low liquid transfer speeds have prevented the useful 96-well plate implementation of FASP as a widely used membrane-based proteomic sample processing method. We validated our approach on whole-cell lysate and urine and cerebrospinal fluid as clinically relevant body fluids. Without compromising peptide and protein identification, our method uses a vacuum manifold and circumvents the need for digest desalting, making our processing method compatible with standard liquid handling robots. In summary, our new method maintains the strengths of FASP and simultaneously overcomes one of the major limitations of FASP without compromising protein identification and quantification. PMID:26223766

  20. Human tear analysis with miniaturized multiplex cytokine assay on “wall-less” 96-well plate

    PubMed Central

    Quah, Joanne; Tong, Louis; Kim, Namyong

    2015-01-01

    Purpose Tears are a particularly limited body fluid and commonly used in the diagnosis of patients who have ocular diseases. A popular method for analysis of ocular inflammation in tears uses Luminex® bead multiplex technology to generate valuable multiple cytokine profile outputs with 25–50 µl tear sample volume. We propose a method for measuring tear cytokines with 5 μl tear sample volume and 80% reduced Luminex reagents compared to previous protocols. Methods Using human tears pooled from 1,000 participants, the DA-Bead-based method running at 5–20 µl volume, using manual pipetting, in conjunction with a magnetic Luminex cytokine (four-plex) panel assay in a 96-well format was performed and validated for tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-1β, and IL-6. Results Upon use of the DA-Bead method at the 5 μl volume with cytokine standards, the concentrations of each of the four cytokines were found to be linear over a range of 3.5–4 log pg/ml with an intra-assay coefficient of variation (CV) ≤5%, inter-assay %CV ≤10%, and accuracy within the 70–130% range. Upon use of a 5 µl healthy pooled tear sample, cytokine concentrations were detected with a precision intra-assay %CV ˂ 20% for IL-6, IFN-γ, or TNF-α or 30.37% with IL-1β. The inter-assay %CV with tears was ≤20.84% for all cytokines. Tear volumes run at 5 μl on DA-Bead produced a similar cytokine expression profile at a 1-month interval and were highly correlated with the larger 10 μl–based tear sample volume cytokine profile with R2 = 0.98. Conclusions DA-Bead assay is highly sensitive and reproducible and has a performance profile that is potentially suitable for use in standard clinical scenarios. Considering the use of as little as 5 µl of assay beads and 5 µl sample, this is also likely to reduce the assay cost significantly and ease diagnosis of patients with ocular diseases. PMID:26539027

  1. Is biofilm removal properly assessed? Comparison of different quantification methods in a 96-well plate system.

    PubMed

    Stiefel, Philipp; Rosenberg, Urs; Schneider, Jana; Mauerhofer, Stefan; Maniura-Weber, Katharina; Ren, Qun

    2016-05-01

    Various methods have been reported to quantify total biofilm or different components of biofilm; however, these methods are often confusedly used, leading to discrepancies and misleading results. In this study, different methods for quantification of biofilm, including those for total biomass, total amount of bacterial cells, viable cell number, and amount of extracellular polymeric substances, were systematically compared in microtiter plates. To evaluate which method is suitable for assessment of biofilm removal and for bacterial killing, biofilm samples were treated with various cleaners possessing removing and/or killing capacities. It was found that most of the methods tested in this study in general exhibited high reproducibility and repeatability. Crystal Violet staining was a simple but reliable method for total biomass quantification. Total bacteria cell numbers could be reliably quantified by the fluorescent DNA-binding dye Acridine Orange. Viable cells could be quantified by either an ATP-based assay or a proliferation assay. Both of these viability methods showed a broad detection range and led to precise measurement. For quantification of proteins in the biofilm, staining with fluorescein isothiocyanate was most suitable. Furthermore, it was revealed that a combination of different methods is required to determine if a cleaner kills or removes biofilm. PMID:26923144

  2. Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin.

    PubMed

    Ho, Tien Yu Jessica; Chan, Chia-Chung; Chan, KinGho; Wang, Yu Chieh; Lin, Jing-Tang; Chang, Cheng-Ming; Chen, Chien-Sheng

    2013-11-15

    We developed a sensitive, simple, inexpensive and rapid bead-based immunoassay platform, composed of liposomal nanovesicle amplification system, Gentamycin sulfate beads and 96-well filtration plates. In the beginning of the assay, Gentamycin sulfate beads, Gentamycin sulfate and Gentamycin specific antibody were incubated in a bottom-sealed 96-well filtration plate. After incubation, washing was done by running washing buffer through the unsealed filtration plate with only gravity and the antibody-Gentamycin bead complexes were retained in the plate. Fluorescent dye-loaded protein G-liposomal nanovesicles were then added to specifically bind to antibodies on the retained beads. After washing unbound nanovesicles, millions of fluorescent dye molecules were released by adding a detergent solution to lyse liposomal nanovesicles. The limit of detection (LOD) of this novel detection platform in TBS and in skim milk were 52.65 ng/mL and 14.16 ng/mL, which are both sufficient for detecting the 200 ng/mL Codex maximum residual level (MRL). The dynamic ranges were both from each of their LODs to 100 μg/mL. The 50% inhibition concentrations (IC50) in TBS and skim milk were 199.66 ng/mL and 360.81 ng/mL, respectively. We also demonstrated the good specificity of this platform by comparing detection results between pure Gentamycin solution and a mixture solution of 6 different antibiotics including Gentamycin in skim milk. The entire assay with 60 samples was conducted within 2h. In sum, this novel biosensing platform not only fulfilled most benefits of magnetic bead-based assays, but also was inexpensive and convenient by replacing the magnetic separation with filtration plate separation. PMID:23728198

  3. A Novel 96well-formatted Micro-gap Plate Enabling Drug Response Profiling on Primary Tumour Samples

    NASA Astrophysics Data System (ADS)

    Ma, Wei-Yuan; Hsiung, Lo-Chang; Wang, Chen-Ho; Chiang, Chi-Ling; Lin, Ching-Hung; Huang, Chiun-Sheng; Wo, Andrew M.

    2015-04-01

    Drug-based treatments are the most widely used interventions for cancer management. Personalized drug response profiling remains inherently challenging with low cell count harvested from tumour sample. We present a 96well-formatted microfluidic plate with built-in micro-gap that preserves up to 99.2% of cells during multiple assay/wash operation and only 9,000 cells needed for a single reagent test (i.e. 1,000 cells per test spot x 3 selected concentration x triplication), enabling drug screening and compatibility with conventional automated workstations. Results with MCF7 and MDA-MB-231 cell lines showed that no statistical significance was found in dose-response between the device and conventional 96-well plate control. Primary tumour samples from breast cancer patients tested in the device also showed good IC50 prediction. With drug screening of primary cancer cells must consider a wide range of scenarios, e.g. suspended/attached cell types and rare/abundant cell availability, the device enables high throughput screening even for suspended cells with low cell count since the signature microfluidic cell-trapping feature ensures cell preservation in a multiple solution exchange protocol.

  4. Development of a fluorimetric multispecies 96-well micro-plate growth test for screening metal toxicity to phytoplankton

    SciTech Connect

    Peterson, H.G.; Ruecker, N.J.; Cantin, I.A.; Nyholm, N.; Dal-Jensen, S.

    1995-12-31

    The rapid and cost-effective screening of industrial waste is an ideal approach to regulations that offer true protection of aquatic habitats. For these tests to be ecologically important protection of large groups of organisms is also essential. This can best be done by testing batteries of species. Photosynthetic organisms compose 99.9% of habitats as well as providing food for higher trophic levels. A test was developed that can accommodate the testing of most phytoplanktonic species irrespective of morphology (unicellular, multicellular, colonial, filamentous). Forty eight to 72 h growth tests were carried out with green algae, diatoms, and cyanobacteria. The algae were incubated with different levels of toxicants in 96-well microplates which were read in a 96-well fluorometric plate reader. Phytoplankton emitting low levels of fluorescence can be incubated with DCMU, which can increase the fluorescent signal 2 to 4 times. The data from the plate reader is transferred to a computer spreadsheet and inhibition levels are automatically calculated. Eleven metal mining wastes from across Canada were tested against this method using the following phytoplanktonic species: Selenastrum, Nannochloris (green algae), Nitzschia (diatom), Microcystis, and Pseudoanabaena (cyanobacteria). These wastes were also screened against Microtox. All wastes were highly toxic to the tested phytoplankton, but only 4 were toxic to Microtox{trademark}.

  5. 96-Well Plate Colorimetric Assay for K(sub i) Determination of (plusmn)-2-Benzylsuccinic Acid, an Inhibitor of Carboxypeptidase A

    ERIC Educational Resources Information Center

    Wentland, Mark P.; Raza, Shaan; Yingtong Gao

    2004-01-01

    An appropriate assay to determine the inhibition potency of carboxypeptidase A (CPA) in 96-well format to illustrate how high throughput screening is used in modern drug discovery to identify bioactive molecules is developed. Efforts in developing a colorimetric 96-well plate assay for determination of the K(sub i) for inhibition of CPA by…

  6. Facile preparation of a photoactivatable surface on a 96-well plate: a versatile and multiplex cell migration assay platform.

    PubMed

    Kamimura, Masao; Scheideler, Olivia; Shimizu, Yoshihisa; Yamamoto, Shota; Yamaguchi, Kazuo; Nakanishi, Jun

    2015-06-01

    Cell migration is an essential cellular activity in various physiological and pathological processes, such as wound healing and cancer metastasis. Therefore, in vitro cell migration assays are important not only for fundamental biological studies but also for evaluating potential drugs that control cell migration activity in medical applications. In this regard, robust control over cell migrating microenvironments is critical for reliable and quantitative analysis as cell migration is highly dependent upon the microenvironments. Here, we developed a facile method for making a commercial glass-bottom 96-well plate photoactivatable for cell adhesion, aiming to develop a versatile and multiplex cell migration assay platform. Cationic poly-d-lysine was adsorbed to the anionic glass surface via electrostatic interactions and, subsequently, functionalized with poly(ethylene glycol) (PEG) bearing a photocleavable reactive group. The initial PEGylated surface is non-cell-adhesive. However, upon near-ultraviolet (UV) irradiation, the photorelease of PEG switches the surface from non-biofouling to cell-adhesive. With this platform, we assayed cell migration in the following procedure: (1) create cell-attaching regions of precise geometries by controlled photoirradiation, (2) seed cells to allow them to attach selectively to the irradiated regions, (3) expose UV light to the remaining PEGylated regions to extend the cell-adhesive area, (4) analyse cell migration using microscopy. Surface modification of the glass surface was characterized by ζ-potential and contact angle measurements. The PEGylated surface showed cell-resistivity and became cell-adhesive upon releasing PEG by near-UV irradiation. The method was applied for parallelly evaluating the effect of model drugs on the migration of epithelial MDCK cells in the multiplexed platform. The dose-response relationship for cytochalasin D treatment on cell migration behavior was successfully evaluated with high

  7. Immobilization of saccharides and peptides on 96-well microtiter plates coated with methyl vinyl ether-maleic anhydride copolymer.

    PubMed

    Satoh, A; Kojima, K; Koyama, T; Ogawa, H; Matsumoto, I

    1998-06-15

    We have previously reported a method to immobilize protein ligands on microtiter plates coated with methyl vinyl ether-maleic anhydride copolymer (MMAC) [Isosaki, K., et al. (1992) J. Chromatogr. 597, 123-128]. In this study, we improved the MMAC method to efficiently immobilize not only small ligands such as peptides and oligosaccharides, which could not be efficiently immobilized previously, but also heparin via its reducing end. Amino and hydrazino groups were introduced to MMAC-coated microtiter plate wells by coupling to acid anhydride groups of MMAC with 1,6-hexamethylenediamine and adipic acid dihydrazide, respectively. The amino groups introduced were allowed to react with peptides by use of divalent cross-linkers. Hydrazino groups were allowed to react with formyl groups of saccharides by reductive amination. Peptides and oligosaccharides were immobilized in a dose-dependent manner by these methods. In the case of the angiotensin peptide thus immobilized, the detection limit by monoclonal antibodies was as low as 0.1-1 fmol peptide per well. Application of 20-200 nmol oligosaccharides to the well was sufficient to immobilize and subsequently detect lectins. Furthermore, heparin immobilized on the hydrazinocoated wells was successfully used for the binding assay of annexin IV. PMID:9648659

  8. Collection, storage, and filtration of in vivo study samples using 96-well filter plates to facilitate automated sample preparation and LC/MS/MS analysis.

    PubMed

    Berna, M; Murphy, A T; Wilken, B; Ackermann, B

    2002-03-01

    The benefits of high-throughput bioanalysis within the pharmaceutical industry are well established. One of the most significant bottlenecks in bioanalysis is transferring in vivo-generated study samples from their collection tubes during sample preparation and extraction. In most cases, the plasma samples must be stored frozen prior to analysis, and the freeze/thaw (F/T) process introduces thrombin clots that are capable of plugging pipets and automated liquid-transfer systems. A new approach to dealing with this problem involves the use of Ansys Captiva 96-well 20-microm polypropylene filter plates to collect, store frozen, and filter plasma samples prior to bioanalysis. The samples are collected from the test subjects, and the corresponding plasma samples are placed directly into the wells of the filter plate. Two Duoseal (patent pending) covers are used to seal the top and bottom of the plate, and the plate is stored at down to -70 degrees C. Prior to sample analysis, the seals are removed and the plate is placed in a 96-well SPE manifold. As the plasma thaws, it passes (by gravity or mild vacuum) through the polypropylene filter into a 96-well collection plate. A multichannel pipet or automated liquid-transfer system is used to transfer sample aliquots without fear of plugging. A significant advantage of this approach is that, unlike other methods, issues related to incomplete pipetting are virtually eliminated. The entire process is rapid since thawing and filtering take place simultaneously, and if a second F/T cycle is required for reanalysis, it is not necessary to refilter the samples (additional clotting was not observed after three F/T cycles). This technique was tested using monkey, rat, and dog plasma and sodium heparin and EDTA anticoagulants. To assess the possibility of nonspecific binding to the polypropylene filter, a variety of drug candidates from diverse drug classes were studied. Validation data generated for two Lilly compounds from distinct

  9. Quantum dot immunoassays in renewable surface column and 96-well plate formats for the fluorescence detection of Botulinum neurotoxin using high-affinity antibodies

    PubMed Central

    Grate, Jay W.; Tyler, Abby; Ozanich, Richard M.; Miller, Keith D.; Lou, Jianlong; Marks, James D.; Bruckner-Lea, Cynthia J.

    2009-01-01

    A fluorescence sandwich immunoassay using high affinity antibodies and quantum dot (QD) reporters has been developed for detection of botulinum neurotoxin serotype A (BoNT/A) using a nontoxic recombinant fragment of the holotoxin (BoNT/A-HC-fragment) as a structurally valid simulant for the full toxin molecule. The antibodies used, AR4 and RAZ1, bind to nonoverlapping epitopes present on both the full toxin and on the recombinant fragment. In one format, the immunoassay is carried out in a 96-well plate with detection in a standard plate reader using AR4 as the capture antibody and QD-coupled RAZ1 as the reporter. Detection to 31 pM with a total incubation time of 3 hours was demonstrated. In a second format, the AR4 capture antibody was coupled to Sepharose beads, and the reactions were carried out in microcentrifuge tubes with an incubation time of 1 hour. The beads were subsequently captured and concentrated in a rotating rod “renewable surface” flow cell equipped with a fiber optic system for fluorescence measurements. In PBS buffer, the BoNT/A-HC-fragment was detected to concentrations as low as 5 pM using the fluidic measurement approach. PMID:19643593

  10. Quantum dot immunoassays in renewable surface column and 96-well plate formats for the fluorescence detection of Botulinum neurotoxin using high-affinity antibodies

    SciTech Connect

    Warner, Marvin G.; Grate, Jay W.; Tyler, Abby J.; Ozanich, Richard M.; Miller, Keith D.; Lou, Jianlong; Marks, James D.; Bruckner-Lea, Cindy J.

    2009-09-01

    A fluorescence sandwich immunoassay using high affinity antibodies and quantum dot (QD) reporters has been developed for detection of botulinum toxin serotype A (BoNT/A). For the development of the assay, a nontoxic recombinant fragment of the holotoxin (BoNT/A-HC-fragment) has been used as a structurally valid simulant for the full toxin molecule. The antibodies used, AR4 and RAZ1, bind to nonoverlapping epitopes present on both the full toxin and on the recombinant fragment. In one format, the immunoassay is carried out in a 96-well plate with detection in a standard plate reader. Detection down to 31 pM of the BoNT/Hc-fragment was demonstrated with a total incubation time of 3 hours, using AR4 as the capture antibody and QD-coupled RAZ1 as the reporter. In a second format, the AR4 capture antibody was coupled to Sepharose beads, and the immunochemical reactions were carried out in microcentrifuge tubes with an incubation time of 1 hour. These beads were subsequently captured and concentrated in a rotating rod “renewable surface” flow cell as part of a sequential injection fluidic system. This flow cell was equipped with a fiber optic system for fluorescence measurements. In PBS buffer solution matrix, the BoNT/A-HC-fragment was detected to concentrations as low as 5 pM using the fluidic measurement approach.

  11. Identification of photosynthesis inhibitors of pelagic marine algae using 96-well plate microfractionation for enhanced throughput in effect-directed analysis.

    PubMed

    Booij, Petra; Vethaak, A Dick; Leonards, Pim E G; Sjollema, Sascha B; Kool, Jeroen; de Voogt, Pim; Lamoree, Marja H

    2014-07-15

    Because of large-scale production and use of an increasing diversity of chemicals in modern society, estuarine and coastal waters may be contaminated with numerous substances. Some of these compounds have the potential to affect microalgae at the base of the pelagic food chain. Therefore, we identified the main chemical stressors that negatively affect the effective photosystem II efficiency (ϕPSII) in marine microalgae of the Dutch estuarine and coastal waters. An enhanced effect-directed analysis (EDA) was carried out by combining reversed-phase ultra performance liquid chromatography fractionation of extracts from passive samplers, followed by effect assessment using the pulse amplitude modulation fluorometry assay and chemical analysis of biologically active fractions using high-resolution mass spectrometry. This study focuses on a novel microfractionation technique using 96-well plates to enhance throughput in EDA, structure elucidation, and the analytical and effect confirmation of the compounds that are identified. Although there are numerous unknown compounds present in estuarine and coastal waters, our EDA study shows that atrazine, diuron, irgarol, isoproturon, terbutryn, and terbutylazine are the main contributors to the observed effect on the ϕPSII of marine microalgae. PMID:24926900

  12. High throughput adjustable 96-well plate assay for androgen receptor binding: a practical approach for EDC screening using the chimpanzee AR.

    PubMed

    Hartig, P C; Cardon, M C; Blystone, C R; Gray, L E; Wilson, V S

    2008-09-26

    The issue as to whether natural and man-made chemicals interfere with endocrine function has raised concerns. This interference could be biologically significant even at very low doses if the chemicals interact deleteriously with hormone receptors at low concentrations. Therefore, the United States Environmental Protection Agency (USEPA) Office of Coordination and Policy (OSCP) requested that a nonhuman mammalian androgen receptor binding assay be developed for possible use in their Endocrine Disruptor Screening Program (EDSP). Ideally, this assay would be high throughput, not use animals as a source of receptor protein, easily deployed throughout the scientific community, utilize reagents available to both the public and private sector, and have the potential for future automation. We developed a highly modified 96-well plate assay which meets these criteria. It employs a baculovirus expressed recombinant primate androgen receptor which is publically available and exploits the unique ability of some mammalian androgen receptors to remain biologically active after guanidine hydrochloride (GdnHCl) solubilization. This GdnHCl treated receptor remains soluble and requires no additional purification prior to use. We provide a very detailed description of the assay protocol itself, and similarly detailed method for producing and solubilizing the receptor. PMID:18691642

  13. A novel colorimetric assay of β-D-glucans in basidiomycete strains by alcian blue dye in a 96-well microtiter plate.

    PubMed

    C Semedo, Magda; Karmali, Amin; Fonseca, Luís

    2015-01-01

    Basidiomycete strains synthesize several types of β-d-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these β-d-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these β-d-glucans present some disadvantages, a novel colorimetric assay method for β-d-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (∼14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of β-d-glucans in the range of 0-0.8 μg, with a slope of 44.15 × 10(-2) and a limit of detection of 0.017 μg/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for β-1,3-d-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red. β-d-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify β-d-glucans from other biological sources. PMID:26317582

  14. Development and application of a high-throughput sample cleanup process based on 96-well plate for simultaneous determination of 16 steroids in biological matrices using liquid chromatography-triple quadrupole mass spectrometry.

    PubMed

    Luo, Guanzhong; Li, Youxin; Bao, James J

    2016-02-01

    A novel high-throughput sample pretreatment system was developed by the integration of protein precipitation (PP), phospholipid removal (PPR), and hollow fiber liquid-phase microextraction (HF-LPME) into two simple 96-well plates and a matching 96-grid lid. With this system, 16 steroids were separated from biological matrices of plasma, milk, and urine and analyzed by liquid chromatography-triple quadrupole mass spectrometry. In the tandem sample cleanup process, the prepositive PP and PPR step preliminarily removed some of the interferences from the biological matrices. The following HF-LPME step kept the residual interference out of the hollow fiber and enriched the steroids in the hollow fiber to achieve high sensitivity. By a series of method optimizations, acetonitrile was chosen as the crash solvent for PP and PPR. A mixture of octanol and toluene (1:1 v/v) was used as the acceptor phase for HF-LPME. The extraction was conducted at 80 rpm for 50 min in a donor phase containing 1 mL 20% sodium chloride at 25 °C. Under these conditions, the limits of detection for the 16 steroids were 3.6-300.0 pg(.)mL(-1) in plasma, 3.0-270.0 pg·mL(-1) in milk, and 2.2-210.0 pg(.)mL(-1) in urine. The recoveries of the 16 steroids were 81.9-97.9% in plasma (relative standard deviation 1.0-8.0%), 80.6-97.7% in milk (relative standard deviation 0.8-5.4%), and 87.3-98.7% in urine (relative standard deviation 1.0-4.9%). Further, the integrated 96-well platform of PP, PPR, and HF-LPME enabled us to run this assay in an automatic and high-throughput fashion. The reliability of the method was further corroborated by evaluation of its applicability in plasma and urine samples from volunteers and fresh bovine milk from local dairy enterprises. PMID:26738495

  15. Location dependent biases in automatic 96-well microplate readers.

    PubMed

    Harrison, R O; Hammock, B D

    1988-01-01

    Procedures performed in 96-well microplates and quantitated by automatic readers assume instruments to be precise, accurate, and free of well location dependent bias. Instrument specifications generally focus on precision and accuracy without specifically addressing biases which are dependent on well location. These biases appear to be meniscus dependent and can be demonstrated in varying degrees in automatic readers of many designs by using a reverse plate wet test, which compares repeated readings of a dye loaded plate in normal and reversed positions. This test analyzes differences between readings and is, therefore, independent of pipetting error or other experimental variables such as protein binding or immunoassay variability. Different plates increased or decreased the magnitude of observed errors but did not themselves cause the errors measured by the reverse plate wet test. Error patterns were consistent for each reader and varied widely among the 16 instruments tested. Only 4 of 16 instruments passed an existing manufacturer's specification for precision, and only one of the 16 readers tested passed a similar specification for accuracy. The severest location dependent bias was found in an instrument which exhibited excellent repeatability and consistently passed its built-in diagnostic tests. One reader with significant bias was returned to the manufacturer for routine service and calibration, but it was not demonstrably improved. The reverse plate wet test is an extremely useful diagnostic tool for quality control at all stages of instrument manufacture and use. PMID:3069835

  16. Establishment and validation of a method for multi-dose irradiation of cells in 96-well microplates

    SciTech Connect

    Abatzoglou, Ioannis; Zois, Christos E.; Pouliliou, Stamatia

    2013-02-15

    Highlights: ► We established a method for multi-dose irradiation of cell cultures within a 96-well plate. ► Equations to adjust to preferable dose levels are produced and provided. ► Up to eight different dose levels can be tested in one microplate. ► This method results in fast and reliable estimation of radiation dose–response curves. -- Abstract: Microplates are useful tools in chemistry, biotechnology and molecular biology. In radiobiology research, these can be also applied to assess the effect of a certain radiation dose delivered to the whole microplate, to test radio-sensitivity, radio-sensitization or radio-protection. Whether different radiation doses can be accurately applied to a single 96-well plate to further facilitate and accelerated research by one hand and spare funds on the other, is a question dealt in the current paper. Following repeated ion-chamber, TLD and radiotherapy planning dosimetry we established a method for multi-dose irradiation of cell cultures within a 96-well plate, which allows an accurate delivery of desired doses in sequential columns of the microplate. Up to eight different dose levels can be tested in one microplate. This method results in fast and reliable estimation of radiation dose–response curves.

  17. 96-well format-based microfluidic platform for parallel interconnection of multiple multicellular spheroids.

    PubMed

    Kim, Jin-Young; Fluri, David A; Kelm, Jens M; Hierlemann, Andreas; Frey, Olivier

    2015-06-01

    In this article, we present a microfluidic platform, compatible with conventional 96-well formats, that enables facile and parallelized culturing and testing of spherical microtissues in a standard incubator. The platform can accommodate multiple microtissues (up to 66) of different cell types, formed externally by using the hanging-drop method, and enables microtissue interconnection through microfluidic channels for continuous media perfusion or dosage of substances. The platform contains 11 separate channels, and each channel has six tissue compartments. Primary rat liver tissues were cultured over 8 days, and multiple tumor tissues (HCT116) were exposed to various concentrations of 5-fluorouracil for platform characterization. PMID:25524491

  18. Automation of cell-based drug absorption assays in 96-well format using permeable support systems.

    PubMed

    Larson, Brad; Banks, Peter; Sherman, Hilary; Rothenberg, Mark

    2012-06-01

    Cell-based drug absorption assays, such as Caco-2 and MDCK-MDR1, are an essential component of lead compound ADME/Tox testing. The permeability and transport data they provide can determine whether a compound continues in the drug discovery process. Current methods typically incorporate 24-well microplates and are performed manually. Yet the need to generate absorption data earlier in the drug discovery process, on an increasing number of compounds, is driving the use of higher density plates. A simple, more efficient process that incorporates 96-well permeable supports and proper instrumentation in an automated process provides more reproducible data compared to manual methods. Here we demonstrate the ability to perform drug permeability and transport assays using Caco-2 or MDCKII-MDR1 cells. The assay procedure was automated in a 96-well format, including cell seeding, media and buffer exchanges, compound dispense, and sample removal using simple robotic instrumentation. Cell monolayer integrity was confirmed via transepithelial electrical resistance and Lucifer yellow measurements. Proper cell function was validated by analyzing apical-to-basolateral and basolateral-to-apical movement of rhodamine 123, a known P-glycoprotein substrate. Apparent permeability and efflux data demonstrate how the automated procedure provides a less variable method than manual processing, and delivers a more accurate assessment of a compound's absorption characteristics. PMID:22357561

  19. High throughput preparation of fly genomic DNA in 96-well format using a paint-shaker.

    PubMed

    Lang, Michael; Nagy, Olga; Lang, Claus; Orgogozo, Virginie

    2015-01-01

    Sample homogenization is an essential step for genomic DNA extraction, with multiple downstream applications in Molecular Biology. Genotyping hundreds or thousands of samples requires an automation of this homogenization step, and high throughput homogenizer equipment currently costs 7000 euros or more. We present an apparatus for homogenization of individual Drosophila adult flies in 96-well micro-titer dishes, which was built from a small portable paint-shaker (F5 portable paint-shaker, Ushake). Single flies are disrupted in each well that contains extraction buffer and a 4-mm metal ball. Our apparatus can hold up to five 96-well micro-titer plates. Construction of the homogenizer apparatus takes about 3-4 days, and all equipment can be obtained from a home improvement store. The total material cost is approximately 700 euros including the paint-shaker. We tested the performance of our apparatus using the ZR-96 Quick-gDNA™ kit (Zymo Research) homogenization buffer and achieved nearly complete tissue homogenization after 15 minutes of shaking. PCR tests did not detect any cross contamination between samples of neighboring wells. We obtained on average 138 ng of genomic DNA per fly, and DNA quality was adequate for standard PCR applications. In principle, our tissue homogenizer can be used for isolation of DNA suitable for library production and high throughput genotyping by Multiplexed Shotgun Genotyping (MSG), as well as RNA isolation from single flies. The sample adapter can also hold and shake other items, such as centrifuge tubes (15-50 mL) or small bottles. PMID:26818699

  20. High throughput preparation of fly genomic DNA in 96-well format using a paint-shaker

    PubMed Central

    Lang, Michael; Nagy, Olga; Lang, Claus; Orgogozo, Virginie

    2015-01-01

    Sample homogenization is an essential step for genomic DNA extraction, with multiple downstream applications in Molecular Biology. Genotyping hundreds or thousands of samples requires an automation of this homogenization step, and high throughput homogenizer equipment currently costs 7000 euros or more. We present an apparatus for homogenization of individual Drosophila adult flies in 96-well micro-titer dishes, which was built from a small portable paint-shaker (F5 portable paint-shaker, Ushake). Single flies are disrupted in each well that contains extraction buffer and a 4-mm metal ball. Our apparatus can hold up to five 96-well micro-titer plates. Construction of the homogenizer apparatus takes about 3–4 days, and all equipment can be obtained from a home improvement store. The total material cost is approximately 700 euros including the paint-shaker. We tested the performance of our apparatus using the ZR-96 Quick-gDNA™ kit (Zymo Research) homogenization buffer and achieved nearly complete tissue homogenization after 15 minutes of shaking. PCR tests did not detect any cross contamination between samples of neighboring wells. We obtained on average 138 ng of genomic DNA per fly, and DNA quality was adequate for standard PCR applications. In principle, our tissue homogenizer can be used for isolation of DNA suitable for library production and high throughput genotyping by Multiplexed Shotgun Genotyping (MSG), as well as RNA isolation from single flies. The sample adapter can also hold and shake other items, such as centrifuge tubes (15–50 mL) or small bottles. PMID:26818699

  1. Determining cereal starch amylose content using a dual wavelength iodine binding 96 well plate assay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Studying these large...

  2. Development of a 96-well plate iodine binding assay for amylose content determination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cereal starch amylose/amylopectin (AM/AP) ratios are critical in functional properties for food and industrial applications. Conventional methods for the determination of AM/AP of cereal starches are very time consuming and labor intensive making it very difficult to screen large sample sets. Stud...

  3. Folate content in fresh-cut vegetable packed products by 96-well microtiter plate microbiological assay.

    PubMed

    Fajardo, Violeta; Alonso-Aperte, Elena; Varela-Moreiras, Gregorio

    2015-02-15

    Ready-to-eat foods have nowadays become a significant portion of the diet. Accordingly, nutritional composition of these food categories should be well-known, in particular its folate content. However, there is a broad lack of folate data in food composition tables and databases. A total of 21 fresh-cut vegetable and fruit packed products were analysed for total folate (TF) content using a validated method that relies on the folate-dependent growth of chloramphenicol-resistant Lactobacillus casei subspecies rhamnosus (NCIMB 10463). Mean TF content ranged from 10.0 to 140.9μg/100g for the different matrices on a fresh weight basis. Higher TF quantity, 140.9-70.1μg/100g, was found in spinach, rocket, watercress, chard and broccoli. Significant differences were observed between available data for fresh vegetables and fruits from food composition tables or databases and the analysed results for fresh-cut packed products. Supplied data support the potential of folate-rich fresh-cut ready-to-eat vegetables to increase folate intake significantly. PMID:25236228

  4. HIGHLY SENSITIVE ASSAY FOR ANTICHOLINESTERASE COMPOUNDS USING 96 WELL PLATE FORMAT

    EPA Science Inventory

    One of the approaches for reducing uncertainties in the assessment of human exposure is to better characterize concentrations of hazardous compounds that may be present in our immediate environment. A significant limitation to this approach, however, is that sampling and labora...

  5. Protocol: high throughput silica-based purification of RNA from Arabidopsis seedlings in a 96-well format

    PubMed Central

    2011-01-01

    The increasing popularity of systems-based approaches to plant research has resulted in a demand for high throughput (HTP) methods to be developed. RNA extraction from multiple samples in an experiment is a significant bottleneck in performing systems-level genomic studies. Therefore we have established a high throughput method of RNA extraction from Arabidopsis thaliana to facilitate gene expression studies in this widely used plant model. We present optimised manual and automated protocols for the extraction of total RNA from 9-day-old Arabidopsis seedlings in a 96 well plate format using silica membrane-based methodology. Consistent and reproducible yields of high quality RNA are isolated averaging 8.9 μg total RNA per sample (~20 mg plant tissue). The purified RNA is suitable for subsequent qPCR analysis of the expression of over 500 genes in triplicate from each sample. Using the automated procedure, 192 samples (2 × 96 well plates) can easily be fully processed (samples homogenised, RNA purified and quantified) in less than half a day. Additionally we demonstrate that plant samples can be stored in RNAlater at -20°C (but not 4°C) for 10 months prior to extraction with no significant effect on RNA yield or quality. Additionally, disrupted samples can be stored in the lysis buffer at -20°C for at least 6 months prior to completion of the extraction procedure providing a flexible sampling and storage scheme to facilitate complex time series experiments. PMID:22136293

  6. Multiwell cell culture plate format with integrated microfluidic perfusion system

    NASA Astrophysics Data System (ADS)

    Domansky, Karel; Inman, Walker; Serdy, Jim; Griffith, Linda G.

    2006-01-01

    A new cell culture analog has been developed. It is based on the standard multiwell cell culture plate format but it provides perfused three-dimensional cell culture capability. The new capability is achieved by integrating microfluidic valves and pumps into the plate. The system provides a means to conduct high throughput assays for target validation and predictive toxicology in the drug discovery and development process. It can be also used for evaluation of long-term exposure to drugs or environmental agents or as a model to study viral hepatitis, cancer metastasis, and other diseases and pathological conditions.

  7. Technique for the cultural examination of urine using a single plate (the Macba plate).

    PubMed Central

    Naylor, G. R.

    1980-01-01

    This technique for the cultural examination of urine provides, within 24 hours, using a single Petri dish, the colonial appearance of any bacteria present on MacConkey agar and on blood agar, a rough colony count of the bacteria present, their sensitivity to four chemotherapeutic agents, isolated colonies from which further sensitivity tests can be done if necessary and the opportunity to scrutinize contaminating organisms that, even though present in only small numbers, may be of help in diagnostic problems. Images Plate 2 Plate 1 PMID:6762392

  8. Protein farnesyltransferase: measurement of enzymatic activity in 96-well format using TopCount microplate scintillation counting technology.

    PubMed

    Harwood, H J

    1995-04-10

    Protein farnesyltransferase (PFT) catalyzes the transfer of the farnesyl group of farnesyl pyrophosphate (FPP) to proteins ending with a carboxy-terminal CAAX motif, forming a thioether linkage to the cysteine residue of the protein. A method is described herein for measurement of PFT activity in 96-well format using TopCount microplate scintillation counting technology. This method has the advantages of requiring only a single transfer from reaction vessels or wells of a 96-well reaction plate to the filtration wells of a 96-well Packard UniFilter GF/B filtration plate following acid precipitation and of allowing liquid scintillation counting to be conducted directly in the filtration plate without the need for either removal of the filter from the plate or transfer of the filter to liquid scintillation vials prior to radioactivity assessment. Using rat brain cytosol as the source of PFT, H-ras as the source of farnesyl acceptor protein, and [1-3H]FPP as the farnesyl donor, the incorporation of radiolabeled farnesyl residues into H-ras was found to be a linear function of both the time of incubation at 37 degrees C (up to 75 min) and the concentration of rat brain cytosolic protein present during incubation (up to 40 micrograms protein), and to be dependent on the concentration of H-ras (Km = 1.1 microM) and FPP (Km = 0.6 microM) present in the incubation reaction. In the presence of 4 microM H-ras, 0.5 microM FPP, 4 mM MgCl2, and 20 microM ZnCl2, the specific activity of rat brain cytosolic PFT measured using this methodology was 0.253 +/- 0.036 (SD; n = 30) pmol H-ras farnesylated per minute of incubation at 37 degrees C per milligram cytosolic protein. The signal-to-noise ratio for H-ras farnesylation using this methodology averaged 25 relative to incubation in the absence of H-ras (background farnesylation of cytosolic proteins) and 50 relative to incubation in the absence of both H-ras and rat brain cytosol (background filter associated radioactivity

  9. Culture of bovine embryos on a polydimethylsiloxane (PDMS) microwell plate.

    PubMed

    Akagi, Satoshi; Hosoe, Misa; Matsukawa, Kazutsugu; Ichikawa, Akihiko; Tanikawa, Tamio; Takahashi, Seiya

    2010-08-01

    We fabricated a polydimethylsiloxane (PDMS)-based microwell plate (PDMS-MP) containing 100 microwells with a rounded bottom and examined whether it can be used for culture of individual in vitro fertilized (IVF) embryos or parthenogenetically activated zona-free embryos in cattle. In Experiment 1, we examined the in vitro developmental ability of IVF embryos cultured individually on PDMS-MP. After IVF, 20 embryos were transferred into 100 microl drops on PDMS-MP and cultured individually in each well of PDMS-MP (PDMS group). After 7 days of culture, the embryos in the PDMS group developed to the blastocyst stage at the same rate of those in the control group cultured in a group of 20 embryos without PDMS-MP. There were no differences in total number of cells and the ratio of inner cell mass to total cells between the PDMS and control groups. In Experiment 2, we examined the in vitro developmental ability of parthenogenetically activated zona-free bovine embryos cultured individually on PDMS-MP. The zona-free embryos were cultured individually in each well of a PDMS-MP or in each well produced by pressing a darning needle onto the bottom of a culture dish (WOW group). After 7 days of culture, the blastocyst formation rate and cell number of blastocysts in the PDMS group did not differ from those of the zona-intact embryos in the control group. Also, there were no differences in the blastocyst formation rate and cell number of blastocysts between the WOW and PDMS groups. These results suggest that the culture system using PDMS-MP is useful for individual embryos or zona-free embryos in cattle. PMID:20484872

  10. A novel 96-well gel-based assay for determining antifungal activity against filamentous fungi.

    PubMed

    Troskie, Anscha Mari; Vlok, Nicolas Maré; Rautenbach, Marina

    2012-12-01

    In recent years the global rise in antibiotic resistance and environmental consciousness lead to a renewed fervour to find and develop novel antibiotics, including antifungals. However, the influence of the environment on antifungal activity is often disregarded and many in vitro assays may cause the activity of certain antifungals to be overestimated or underestimated. The general antifungal test assays that are economically accessible to the majority of scientists primarily rely on visual examination or on spectrophotometric analysis. The effect of certain morphogenic antifungals, which may lead to hyperbranching of filamentous fungi, unfortunately renders these methods unreliable. To minimise the difficulties experienced as a result of hyperbranching, we developed a straightforward, economical 96-well gel-based method, independent of spectrophotometric analysis, for highly repeatable determination of antifungal activity. For the calculation of inhibition parameters, this method relies on the visualisation of assay results by digitisation. The antifungal activity results from our novel micro-gel dilution assay are comparable to that of the micro-broth dilution assay used as standard reference test of The Clinical and Laboratory Standard Institute. Furthermore, our economical assay is multifunctional as it permits microscopic analysis of the preserved assay results, as well as rendering highly reliable data. PMID:23089670

  11. A 96-well electrochemical method for the screening of enzymatic activities.

    PubMed

    Abdellaoui, Sofiène; Noiriel, Alexandre; Henkens, Robert; Bonaventura, Celia; Blum, Loïc J; Doumèche, Bastien

    2013-04-01

    The rapid electrochemical screening of enzyme activities in bioelectronics is still a challenging issue. In order to solve this problem, we propose to use a 96-well electrochemical assay. This system is composed of 96 screen-printed electrodes on a printed circuit board adapted from a commercial system (carbon is used as the working electrode and silver chloride as the counter/reference electrode). The associated device allows for the measurements on the 96 electrodes to be performed within a few seconds. In this work, we demonstrate the validity of the screening method with the commercial laccase from the fungus Trametes versicolor. The signal-to-noise ratio (S/N) is found to be the best way to analyze the electrochemical signals. The S/N follows a saturation-like mechanism with a dynamic linear range of two decades ranging from 0.5 to 75 ng of laccase (corresponding to enzymatic activities from 62 × 10(-6) to 9.37 × 10(-3) μmol min(-1)) and a sensitivity of 3027 μg(-1) at +100 mV versus Ag/AgCl. Laccase inhibitors (azide and fluoride anions), pH optima, and interfering molecules could also be identified within a few minutes. PMID:23461701

  12. Oxygen transfer rates in shaken culture vessels from Fernbach flasks to microtiter plates.

    PubMed

    Running, Jeffrey A; Bansal, Karan

    2016-08-01

    By a sulfite oxidation method, oxygen transfer rates (OTRs) were determined in 11 types of culture vessels from 2.8-L Fernbach (FB) flasks to 96-, 48-, and 24-well square deepwell microtiter plates (MTPs). OTRs ranged from 140 mM/h in 250-mL Ultrayield™ flasks shaken at 300 rpm with a 50 mm diameter shaker throw to 5 mM/h in unbaffled FBs shaken at 200 rpm with a 25 mm throw. Baffles in FBs increased OTRs 6-12-fold under various shaking conditions, and up to five-fold in 250-mL flasks, depending on the type of baffles. Corner-baffling was superior to bottom-baffling in glass, 250-mL flasks. In MTPs, OTRs increased with increasing well size and decreasing fill volume. At 50 mm throw and 300 rpm, 24-well MTPs had OTRs comparable to corner-baffled, 250-mL flasks (∼100 mM/h). The OTRs in unbaffled flasks were relatively insensitive to shaking conditions, increasing less than two-fold between the most modest and the most vigorous conditions. There was no consistency across vessels as to whether the alternate incubation conditions of 70 mm throw and 250 rpm produced higher OTRs than the 50 mm throw and 300 rpm regimen. No increase in OTR was seen in any MTP when the cover hole diameter was increased beyond 4.5 mm. OTRs decreased as viscosity increased, falling smoothly in unbaffled flasks and 24-well MTPs, but 48-well and 96-well MTPs showed precipitous OTR drops as viscosity increased. Matching the OTRs of screening vessels to the oxygen uptake rates of microbial cultures can greatly reduce the number of false positive strains that are forwarded from microbial screens. Biotechnol. Bioeng. 2016;113: 1729-1735. © 2016 Wiley Periodicals, Inc. PMID:26806816

  13. An Assessment of Cell Culture Plate Surface Chemistry for in Vitro Studies of Tissue Engineering Scaffolds

    PubMed Central

    Röder, Alexander; García-Gareta, Elena; Theodoropoulos, Christina; Ristovski, Nikola; Blackwood, Keith A.; Woodruff, Maria A.

    2015-01-01

    The use of biopolymers as a three dimensional (3D) support structure for cell growth is a leading tissue engineering approach in regenerative medicine. Achieving consistent cell seeding and uniform cell distribution throughout 3D scaffold culture in vitro is an ongoing challenge. Traditionally, 3D scaffolds are cultured within tissue culture plates to enable reproducible cell seeding and ease of culture media change. In this study, we compared two different well-plates with different surface properties to assess whether seeding efficiencies and cell growth on 3D scaffolds were affected. Cell attachment and growth of murine calvarial osteoblast (MC3T3-E1) cells within a melt-electrospun poly-ε-caprolactone scaffold were assessed when cultured in either “low-adhesive” non-treated or corona discharged-treated well-plates. Increased cell adhesion was observed on the scaffold placed in the surface treated culture plates compared to the scaffold in the non-treated plates 24 h after seeding, although it was not significant. However, higher cell metabolic activity was observed on the bases of all well-plates than on the scaffold, except for day 21, well metabolic activity was higher in the scaffold contained in non-treated plate than the base. These results indicate that there is no advantage in using non-treated plates to improve initial cell seeding in 3D polymeric tissue engineering scaffolds, however non-treated plates may provide an improved metabolic environment for long-term studies. PMID:26703748

  14. An Assessment of Cell Culture Plate Surface Chemistry for in Vitro Studies of Tissue Engineering Scaffolds.

    PubMed

    Röder, Alexander; García-Gareta, Elena; Theodoropoulos, Christina; Ristovski, Nikola; Blackwood, Keith A; Woodruff, Maria A

    2015-01-01

    The use of biopolymers as a three dimensional (3D) support structure for cell growth is a leading tissue engineering approach in regenerative medicine. Achieving consistent cell seeding and uniform cell distribution throughout 3D scaffold culture in vitro is an ongoing challenge. Traditionally, 3D scaffolds are cultured within tissue culture plates to enable reproducible cell seeding and ease of culture media change. In this study, we compared two different well-plates with different surface properties to assess whether seeding efficiencies and cell growth on 3D scaffolds were affected. Cell attachment and growth of murine calvarial osteoblast (MC3T3-E1) cells within a melt-electrospun poly-ε-caprolactone scaffold were assessed when cultured in either "low-adhesive" non-treated or corona discharged-treated well-plates. Increased cell adhesion was observed on the scaffold placed in the surface treated culture plates compared to the scaffold in the non-treated plates 24 h after seeding, although it was not significant. However, higher cell metabolic activity was observed on the bases of all well-plates than on the scaffold, except for day 21, well metabolic activity was higher in the scaffold contained in non-treated plate than the base. These results indicate that there is no advantage in using non-treated plates to improve initial cell seeding in 3D polymeric tissue engineering scaffolds, however non-treated plates may provide an improved metabolic environment for long-term studies. PMID:26703748

  15. Development of a novel 96-well format for liquid-liquid microextraction and its application in the HPLC analysis of biological samples.

    PubMed

    Borijihan, Guirong; Li, Youxin; Gao, Jianguo; Bao, James J

    2014-05-01

    A novel 96-well liquid-liquid microextraction system combined with modern HPLC was developed and used for the simultaneous analysis of 96 biological samples. The system made use of hollow fibers, a 96-well plate, and a plastic base with a center hole and a side hole. One end of the hollow fiber was sealed, while the other end was attached to one of the holes positioned at the center for the plastic base. The needle was inserted into the liquid from inside or outside of the hollow fiber through the center or the side holes, respectively. The system was tested with plasma samples containing three compounds, acidic indomethacin, neutral dexamethasone, and basic propafenone. Some parameters, such as the kind and dimension of hollow fiber, pH and salt concentration of the donor phase, the selection of organic solvent for the acceptor phase, and the extraction time were investigated. Under the optimization conditions, the Log D and drug concentration of indomethacin, dexamethasone, and propafenone in plasma and urine samples were analyzed. Then, the methodology was validated. The results demonstrated that ng/mL levels could be exactly and rapidly analyzed by our system, which was equipped with an auto-injection sampler, making sample analysis more convenient. PMID:24574156

  16. Efficacy of agar-plate culture in detection of Strongyloides stercoralis infection.

    PubMed

    Arakaki, T; Iwanaga, M; Kinjo, F; Saito, A; Asato, R; Ikeshiro, T

    1990-06-01

    Agar-plate culture of feces using a modified petri dish proved to be highly efficient in the detection of Strongyloides stercoralis infection. Furrows left by S. stercoralis on the agar plate were distinguished readily in size from those left by Necator americanus. PMID:2352073

  17. Effect of thiram on avian growth plate chondrocytes in culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thiram (tetramethyl thiuram disulfide) is a general use pesticide. It causes tibial dyschondroplasia, a cartilage defect in poultry leading to growth plate deformation and lameness. The mechanism of its action on chondrocytes is not understood. Since proteins play significant role in development an...

  18. Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader.

    PubMed

    Schaefer, Jorrit; Jovanovic, Goran; Kotta-Loizou, Ioly; Buck, Martin

    2016-06-15

    Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ß-d-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. PMID:27036618

  19. Single-step method for β-galactosidase assays in Escherichia coli using a 96-well microplate reader

    PubMed Central

    Schaefer, Jorrit; Jovanovic, Goran; Kotta-Loizou, Ioly; Buck, Martin

    2016-01-01

    Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ß-d-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. PMID:27036618

  20. Fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format.

    PubMed

    Rühmann, Broder; Schmid, Jochen; Sieber, Volker

    2014-07-11

    A fast carbohydrate screening platform processible in 96-well format is described. The method is suitable for the determination of various carbohydrates out of complex mixtures as obtained by acidic hydrolysis of carbohydrates polymers. The chromatographic conditions for an efficient separation (12min) and the derivatization process with 1-phenyl-3-methyl-5-pyrazolone (PMP) were optimized for high resolution separation and simultaneous determination of deoxy-, amino-, anhydro-sugars as well as hexoses, pentoses, dimers, uronic acids and degradation products like furfural and hydroxymethylfurfural (HMF). The potential to quantify with UV- and MS-detector in the same range has been demonstrated for 20 different compounds. Finally, the matrix effects of the hydrolysis were positively evaluated. The micro scale hydrolysis and PMP-derivatization without any extraction or drying steps, both in 96-well format, result in a fast and intuitive sample preparation. In combination with a fast liquid chromatography coupled to UV and electrospray ionization ion trap detection (LC-UV-ESI-MS/MS) for the qualification and quantification of various sugars, dimers and degradation products, this method shows great performance in carbohydrate analysis. PMID:24861788

  1. Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay

    PubMed Central

    Lordkipanidzé, Marie; Lowe, Gillian C.; Kirkby, Nicholas S.; Chan, Melissa V.; Lundberg, Martina H.; Morgan, Neil V.; Bem, Danai; Nisar, Shaista P.; Leo, Vincenzo C.; Jones, Matthew L.; Mundell, Stuart J.; Daly, Martina E.; Mumford, Andrew D.; Warner, Timothy D.; Watson, Steve P.; Watson, Steve P.; Mumford, Andrew D.; Mundell, Stuart J.; Gissen, Paul; Daly, Martina E.; Lester, Will; Clark, Justin; Williams, Mike; Motwani, Jayashree; Marshall, Dianne; Nyatanga, Priscilla; Mann, Pat; Kirwan, Julie; Wilde, Jonathan; Dunkley, Tracey; Greenway, April; Makris, Michael; Pavord, Sue; Dattani, Rashesh; Grimley, Gerry Dolan Charlotte; Stokley, Simone; Astwood, Emma; Chang, Cherry; Foros, Merri; Trower, Linda; Thachil, Jecko; Hay, Charlie; Pike, Gill; Will, Andrew; Grainger, John; Foulkes, Matt; Fareh, Mona; Talks, Kate; Biss, Tina; Kesteven, Patrick; Hanley, John; Vowles, Julie; Basey, Lesley; Barnes, Michelle; Collins, Peter; Rayment, Rachel; Alikhan, Raza; Morris, Ana Guerrero Rebecca; Mansell, Dianne; Toh, Cheng Hock; Martlew, Vanessa; Murphy, Elaine; Lachmann, Robin; Rose, Peter; Chapman, Oliver; Lokare, Anand; Marshall, Kathryn; Khan, Naseem; Keeling, David; Giangrande, Paul; Austin, Steve; Bevan, David; Alamelu, Jayanthi

    2014-01-01

    Up to 1% of the population have mild bleeding disorders, but these remain poorly characterized, particularly with regard to the roles of platelets. We have compared the usefulness of Optimul, a 96-well plate-based assay of 7 distinct pathways of platelet activation to characterize inherited platelet defects in comparison with light transmission aggregometry (LTA). Using Optimul and LTA, concentration-response curves were generated for arachidonic acid, ADP, collagen, epinephrine, Thrombin receptor activating-peptide, U46619, and ristocetin in samples from (1) healthy volunteers (n = 50), (2) healthy volunteers treated with antiplatelet agents in vitro (n = 10), and (3) patients with bleeding of unknown origin (n = 65). The assays gave concordant results in 82% of cases (κ = 0.62, P < .0001). Normal platelet function results were particularly predictive (sensitivity, 94%; negative predictive value, 91%), whereas a positive result was not always substantiated by LTA (specificity, 67%; positive predictive value, 77%). The Optimul assay was significantly more sensitive at characterizing defects in the thromboxane pathway, which presented with normal responses with LTA. The Optimul assay is sensitive to mild platelet defects, could be used as a rapid screening assay in patients presenting with bleeding symptoms, and detects changes in platelet function more readily than LTA. This trial was registered at www.isrctn.org as #ISRCTN 77951167. PMID:24408324

  2. Characterization of multiple platelet activation pathways in patients with bleeding as a high-throughput screening option: use of 96-well Optimul assay.

    PubMed

    Lordkipanidzé, Marie; Lowe, Gillian C; Kirkby, Nicholas S; Chan, Melissa V; Lundberg, Martina H; Morgan, Neil V; Bem, Danai; Nisar, Shaista P; Leo, Vincenzo C; Jones, Matthew L; Mundell, Stuart J; Daly, Martina E; Mumford, Andrew D; Warner, Timothy D; Watson, Steve P

    2014-02-20

    Up to 1% of the population have mild bleeding disorders, but these remain poorly characterized, particularly with regard to the roles of platelets. We have compared the usefulness of Optimul, a 96-well plate-based assay of 7 distinct pathways of platelet activation to characterize inherited platelet defects in comparison with light transmission aggregometry (LTA). Using Optimul and LTA, concentration-response curves were generated for arachidonic acid, ADP, collagen, epinephrine, Thrombin receptor activating-peptide, U46619, and ristocetin in samples from (1) healthy volunteers (n = 50), (2) healthy volunteers treated with antiplatelet agents in vitro (n = 10), and (3) patients with bleeding of unknown origin (n = 65). The assays gave concordant results in 82% of cases (κ = 0.62, P < .0001). Normal platelet function results were particularly predictive (sensitivity, 94%; negative predictive value, 91%), whereas a positive result was not always substantiated by LTA (specificity, 67%; positive predictive value, 77%). The Optimul assay was significantly more sensitive at characterizing defects in the thromboxane pathway, which presented with normal responses with LTA. The Optimul assay is sensitive to mild platelet defects, could be used as a rapid screening assay in patients presenting with bleeding symptoms, and detects changes in platelet function more readily than LTA. This trial was registered at www.isrctn.org as #ISRCTN 77951167. PMID:24408324

  3. Development and validation of a 96-well cellular assay for the discovery of ALDH1A1 inhibitors.

    PubMed

    Ming, Wenyu; Ma, Wenzhen; Chen, Lisa H; Volk, Catherine; Michael, Mervyn Dodson; Xu, Yanping; Zhang, Fang; Wang, Xiaojun

    2013-07-01

    Retinoic acid, the active metabolite of vitamin A, plays important roles in various physiological and pathological processes. The two-step production of retinoic acid from vitamin A (retinol) is catalyzed by alcohol dehydrogenases and aldehyde dehydrogenases, which are potential therapeutic targets for numerous diseases, such as obesity, diabetes, and cancer. Currently, the lack of a suitable high-throughput cellular assay hinders efforts to identify therapeutic small molecular inhibitors of aldehyde dehydrogenase, such as ALDH1A1. In this report, we utilized high-content imaging technology and a commercially available cell permeable ALDH substrate to develop a 96-well cellular ALDH1A1 assay. This assay has a robust and sensitive readout and is amenable to automation. With this cellular assay, we identified potent selective ALDH1A1 inhibitors to explore the role of retinoic acid production in various preclinical disease models. PMID:23957476

  4. Evaluation of the Minnesota Easy Culture System II Bi-Plate and Tri-Plate for identification of common mastitis pathogens in milk.

    PubMed

    Royster, E; Godden, S; Goulart, D; Dahlke, A; Rapnicki, P; Timmerman, J

    2014-01-01

    The objective of this study was to validate use of the Minnesota Easy Culture System II Bi-Plate and Tri-Plate (University of Minnesota Laboratory for Udder Health, St. Paul) to identify common mastitis pathogens in milk. A total of 283 quarter and composite milk samples submitted to the University of Minnesota Laboratory for Udder Health during the spring of 2010 were cultured simultaneously using 3 methods: standard laboratory culture (reference method) and the Minnesota Easy Culture System II Bi-Plate and Tri-Plate methods. Bi-Plate and Tri-Plate cultures were incubated for 18 to 24h and interpreted by 2 independent, untrained readers within 5h of each other. An experienced technician completed the standard laboratory culture. For each sample, all 3 study personnel recorded the culture result (yes/no) for each of the following diagnostic categories: no bacterial growth (NG), mixed (2 organisms), contaminated (3 or more organisms), gram-positive (GP), gram-negative (GN), Staphylococcus spp., Streptococcus spp., Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Enterococcus spp., Staphylococcus aureus, coagulase-negative staphylococci, Escherichia coli, Klebsiella spp., and other. For each category, the prevalence, sensitivity, specificity, accuracy, and predictive values of a positive and negative test were calculated, and the agreement between readers and between each reader and the laboratory was assessed. Specificity, overall accuracy, and negative predictive values were generally high (>80%) for the Bi-Plate and Tri-Plate for each category. Sensitivity and positive predictive values were intermediate (>60%) or high (>80%) for the broad categories of NG, GP, GN, Staphylococcus spp. and Streptococcus spp., and for Staph. aureus, but were generally lower (<60%) for other more specific categories. Similarly, interreader agreement (kappa value) was moderate to substantial (40-80%) for the broad categories of NG, GP, GN, Staphylococcus spp

  5. The BUME method: a novel automated chloroform-free 96-well total lipid extraction method for blood plasma[S

    PubMed Central

    Löfgren, Lars; Ståhlman, Marcus; Forsberg, Gun-Britt; Saarinen, Sinikka; Nilsson, Ralf; Hansson, Göran I.

    2012-01-01

    Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profiles of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroform-based reference method. Lipid recoveries were linear from 10–100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by two-phase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, high-throughput BUME method can replace chloroform-based methods, saving both human and environmental resources. PMID:22645248

  6. Microscale 3D collagen cell culture assays in conventional flat-bottom 384-well plates.

    PubMed

    Leung, Brendan M; Moraes, Christopher; Cavnar, Stephen P; Luker, Kathryn E; Luker, Gary D; Takayama, Shuichi

    2015-04-01

    Three-dimensional (3D) culture systems such as cell-laden hydrogels are superior to standard two-dimensional (2D) monolayer cultures for many drug-screening applications. However, their adoption into high-throughput screening (HTS) has been lagging, in part because of the difficulty of incorporating these culture formats into existing robotic liquid handling and imaging infrastructures. Dispensing cell-laden prepolymer solutions into 2D well plates is a potential solution but typically requires large volumes of reagents to avoid evaporation during polymerization, which (1) increases costs, (2) makes drug penetration variable and (3) complicates imaging. Here we describe a technique to efficiently produce 3D microgels using automated liquid-handling systems and standard, nonpatterned, flat-bottomed, 384-well plates. Sub-millimeter-diameter, cell-laden collagen gels are deposited on the bottom of a ~2.5 mm diameter microwell with no concerns about evaporation or meniscus effects at the edges of wells, using aqueous two-phase system patterning. The microscale cell-laden collagen-gel constructs are readily imaged and readily penetrated by drugs. The cytotoxicity of chemotherapeutics was monitored by bioluminescence and demonstrated that 3D cultures confer chemoresistance as compared with similar 2D cultures. Hence, these data demonstrate the importance of culturing cells in 3D to obtain realistic cellular responses. Overall, this system provides a simple and inexpensive method for integrating 3D culture capability into existing HTS infrastructure. PMID:25510473

  7. Microscale 3-D collagen cell culture assays in conventional flat-bottom 384-well plates

    PubMed Central

    Leung, Brendan M.; Moraes, Christopher; Cavnar, Stephen; Luker, Kathryn E.; Luker, Gary D.; Takayama, Shuichi

    2015-01-01

    Three-dimensional culture systems such as cell-laden hydrogels are superior to standard 2-D monolayer cultures for many drug-screening applications. However, their adoption in high throughput screening (HTS) have been lagging, in part due to the difficulty of incorporating these culture formats into existing robotic liquid handling and imaging infrastructures. Dispensing cell-laden pre-polymer solutions into 2-D well-plates is a potential solution, but typically requires large volumes of reagents to avoid evaporation during polymerization, which increases cost, makes drug penetration variable and imaging complex. Here we describe a technique to efficiently produce 3-D ‘microgels’ using automated liquid handling systems and standard, non-patterned, flat-bottomed, 384-well plates. Sub-millimeter-diameter, cell-laden collagen gels are deposited on the bottom of ~2.5 mm-diameter microwell with no concerns over evaporation and meniscus effects at the edges of wells, using aqueous two-phase system patterning. The microscale cell-laden collagen-gel constructs are readily imaged and readily penetrated by drugs. Cytotoxicity of chemotherapeutics were monitored by bioluminescence and demonstrates that 3-D cultures confer chemoresistance, as compared to similar 2-D culture. This data hence, demonstrates the importance of culturing cells in 3-D to obtain realistic cellular responses. Overall, this system provided a simple and inexpensive method for integrating 3-D culture capability into existing HTS infrastructure. PMID:25510473

  8. Determination of suvorexant in human plasma using 96-well liquid-liquid extraction and HPLC with tandem mass spectrometric detection.

    PubMed

    Breidinger, S A; Simpson, R C; Mangin, E; Woolf, E J

    2015-10-01

    A method, using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS), was developed for the determination of suvorexant (MK-4305, Belsomra(®)), a selective dual orexin receptor antagonist for the treatment insomnia, in human plasma over the concentration range of 1-1000ng/mL. Stable isotope labeled (13)C(2)H3-suvorexant was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction, in the 96-well format, of a 100μL plasma sample with methyl t-butyl ether. The compounds were chromatographed under isocratic conditions on a Waters dC18 (50×2.1mm, 3μm) column with a mobile phase consisting of 30/70 (v/v %) 10mM ammonium formate, pH3/acetonitrile at a flow rate of 0.3mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for suvorexant (m/z 451→186) and (13)C(2)H3-suvorexant (m/z 455→190) on an Applied Biosystems API 4000 tandem mass spectrometer was used for quantitation. Intraday assay precision, assessed in six different lots of control plasma, was within 10% CV at all concentrations, while assay accuracy ranged from 95.6 to 105.0% of nominal. Quality control (QC) samples in plasma were stored at -20°C. Initial within day analysis of QCs after one freeze-thaw cycle showed accuracy within 9.5% of nominal with precision (CV) of 6.7% or less. The plasma QC samples were demonstrated to be stable for up to 25 months at -20°C. The method described has been used to support clinical studies during Phase I through III of clinical development. PMID:26343269

  9. Microstructured multi-well plate for three-dimensional packed cell seeding and hepatocyte cell culture

    PubMed Central

    Goral, Vasiliy N.; Au, Sam H.; Faris, Ronald A.; Yuen, Po Ki

    2014-01-01

    In this article, we present a microstructured multi-well plate for enabling three-dimensional (3D) high density seeding and culture of cells through the use of a standard laboratory centrifuge to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro without the addition of animal derived or synthetic matrices or coagulants. Each well has microfeatures on the bottom that are comprised of a series of ditches/open microchannels. The dimensions of the microchannels promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro. After cell seeding with a standard pipette, the microstructured multi-well plates were centrifuged to tightly pack cells inside the ditches in order to enhance cell-cell interactions and induce formation of 3D cellular structures during cell culture. Cell-cell interactions were optimized based on cell packing by considering dimensions of the ditches/open microchannels, orientation of the microstructured multi-well plate during centrifugation, cell seeding density, and the centrifugal force and time. With the optimized cell packing conditions, we demonstrated that after 7 days of cell culture, primary human hepatocytes adhered tightly together to form cord-like structures that resembled 3D tissue-like cellular architecture. Importantly, cell membrane polarity was restored without the addition of animal derived or synthetic matrices or coagulants. PMID:25379107

  10. Uniform electric field generation in circular multi-well culture plates using polymeric inserts

    PubMed Central

    Tsai, Hsieh-Fu; Cheng, Ji-Yen; Chang, Hui-Fang; Yamamoto, Tadashi; Shen, Amy Q.

    2016-01-01

    Applying uniform electric field (EF) in vitro in the physiological range has been achieved in rectangular shaped microchannels. However, in a circular-shaped device, it is difficult to create uniform EF from two electric potentials due to different electrical resistances originated from the length difference between the diameter of the circle and the length of any parallel chord of the bottom circular chamber where cells are cultured. To address this challenge, we develop a three-dimensional (3D) computer-aided designed (CAD) polymeric insert to create uniform EF in circular shaped multi-well culture plates. A uniform EF with a coefficient of variation (CV) of 1.2% in the 6-well plate can be generated with an effective stimulation area percentage of 69.5%. In particular, NIH/3T3 mouse embryonic fibroblast cells are used to validate the performance of the 3D designed Poly(methyl methacrylate) (PMMA) inserts in a circular-shaped 6-well plate. The CAD based inserts can be easily scaled up (i.e., 100 mm dishes) to further increase effective stimulation area percentages, and also be implemented in commercially available cultureware for a wide variety of EF-related research such as EF-cell interaction and tissue regeneration studies. PMID:27193911

  11. Uniform electric field generation in circular multi-well culture plates using polymeric inserts

    NASA Astrophysics Data System (ADS)

    Tsai, Hsieh-Fu; Cheng, Ji-Yen; Chang, Hui-Fang; Yamamoto, Tadashi; Shen, Amy Q.

    2016-05-01

    Applying uniform electric field (EF) in vitro in the physiological range has been achieved in rectangular shaped microchannels. However, in a circular-shaped device, it is difficult to create uniform EF from two electric potentials due to different electrical resistances originated from the length difference between the diameter of the circle and the length of any parallel chord of the bottom circular chamber where cells are cultured. To address this challenge, we develop a three-dimensional (3D) computer-aided designed (CAD) polymeric insert to create uniform EF in circular shaped multi-well culture plates. A uniform EF with a coefficient of variation (CV) of 1.2% in the 6-well plate can be generated with an effective stimulation area percentage of 69.5%. In particular, NIH/3T3 mouse embryonic fibroblast cells are used to validate the performance of the 3D designed Poly(methyl methacrylate) (PMMA) inserts in a circular-shaped 6-well plate. The CAD based inserts can be easily scaled up (i.e., 100 mm dishes) to further increase effective stimulation area percentages, and also be implemented in commercially available cultureware for a wide variety of EF-related research such as EF-cell interaction and tissue regeneration studies.

  12. Uniform electric field generation in circular multi-well culture plates using polymeric inserts.

    PubMed

    Tsai, Hsieh-Fu; Cheng, Ji-Yen; Chang, Hui-Fang; Yamamoto, Tadashi; Shen, Amy Q

    2016-01-01

    Applying uniform electric field (EF) in vitro in the physiological range has been achieved in rectangular shaped microchannels. However, in a circular-shaped device, it is difficult to create uniform EF from two electric potentials due to different electrical resistances originated from the length difference between the diameter of the circle and the length of any parallel chord of the bottom circular chamber where cells are cultured. To address this challenge, we develop a three-dimensional (3D) computer-aided designed (CAD) polymeric insert to create uniform EF in circular shaped multi-well culture plates. A uniform EF with a coefficient of variation (CV) of 1.2% in the 6-well plate can be generated with an effective stimulation area percentage of 69.5%. In particular, NIH/3T3 mouse embryonic fibroblast cells are used to validate the performance of the 3D designed Poly(methyl methacrylate) (PMMA) inserts in a circular-shaped 6-well plate. The CAD based inserts can be easily scaled up (i.e., 100 mm dishes) to further increase effective stimulation area percentages, and also be implemented in commercially available cultureware for a wide variety of EF-related research such as EF-cell interaction and tissue regeneration studies. PMID:27193911

  13. A Chlorhexidine- Agar Plate Culture Medium Protocol to Complement Standard Broth Culture of Mycobacterium tuberculosis

    PubMed Central

    Asmar, Shady; Chatellier, Sonia; Mirande, Caroline; van Belkum, Alex; Canard, Isabelle; Raoult, Didier; Drancourt, Michel

    2016-01-01

    The culture of Mycobacterium tuberculosis using parallel inoculation of a solid culture medium and a liquid broth provides the gold standard for the diagnosis of tuberculosis. Here, we evaluated a chlorhexidine decontamination-MOD9 solid medium protocol versus the standard NALC-NaOH-Bactec 960 MGIT protocol for the diagnosis of pulmonary tuberculosis by culture. Three-hundred clinical specimens comprising 193 sputa, 30 bronchial aspirates, 10 broncho-alveolar lavages, 47 stools, and 20 urines were prospectively submitted for the routine diagnosis of tuberculosis. The contamination rates were 5/300 (1.7%) using the MOD9 protocol and 17/300 (5.7%) with the Bactec protocol, respectively (P < 0.05, Fisher exact test). Of a total of 50 Mycobacterium isolates (48 M. tuberculosis and two Mycobacterium abscessus) were cultured. Out of these 50, 48 (96%) isolates were found using the MOD9 protocol versus 35 (70%) when using the Bactec protocol (P < 0.05, Fisher exact test). The time to positivity was 10.1 ± 3.9 days versus 14.7 ± 7.3 days, respectively, (P < 0.05, Student’s t-test). These data confirmed the usefulness of parallel inoculation of a solid culture medium with broth for the recovery of M. tuberculosis in agreement with current recommendations. More specifically, chlorhexidine decontamination and inoculation of the MOD9 solid medium could be proposed to complement the standard Bactec 960 MGIT broth protocol. PMID:26834733

  14. A Chlorhexidine- Agar Plate Culture Medium Protocol to Complement Standard Broth Culture of Mycobacterium tuberculosis.

    PubMed

    Asmar, Shady; Chatellier, Sonia; Mirande, Caroline; van Belkum, Alex; Canard, Isabelle; Raoult, Didier; Drancourt, Michel

    2016-01-01

    The culture of Mycobacterium tuberculosis using parallel inoculation of a solid culture medium and a liquid broth provides the gold standard for the diagnosis of tuberculosis. Here, we evaluated a chlorhexidine decontamination-MOD9 solid medium protocol versus the standard NALC-NaOH-Bactec 960 MGIT protocol for the diagnosis of pulmonary tuberculosis by culture. Three-hundred clinical specimens comprising 193 sputa, 30 bronchial aspirates, 10 broncho-alveolar lavages, 47 stools, and 20 urines were prospectively submitted for the routine diagnosis of tuberculosis. The contamination rates were 5/300 (1.7%) using the MOD9 protocol and 17/300 (5.7%) with the Bactec protocol, respectively (P < 0.05, Fisher exact test). Of a total of 50 Mycobacterium isolates (48 M. tuberculosis and two Mycobacterium abscessus) were cultured. Out of these 50, 48 (96%) isolates were found using the MOD9 protocol versus 35 (70%) when using the Bactec protocol (P < 0.05, Fisher exact test). The time to positivity was 10.1 ± 3.9 days versus 14.7 ± 7.3 days, respectively, (P < 0.05, Student's t-test). These data confirmed the usefulness of parallel inoculation of a solid culture medium with broth for the recovery of M. tuberculosis in agreement with current recommendations. More specifically, chlorhexidine decontamination and inoculation of the MOD9 solid medium could be proposed to complement the standard Bactec 960 MGIT broth protocol. PMID:26834733

  15. Microfluidic transwell inserts for generation of tissue culture-friendly gradients in well plates

    PubMed Central

    Sip, Christopher G.; Bhattacharjee, Nirveek; Folch, Albert

    2015-01-01

    Gradients of biochemical molecules play a key role in many physiological processes such as axon growth, tissue morphogenesis, and trans-epithelium nutrient transport, as well as in pathophysiological phenomena such as wound healing, immune response, bacterial invasion, and cancer metastasis. In this paper, we report a microfluidic transwell insert for generating quantifiable concentration gradients in a user-friendly and modular format that is compatible with conventional cell cultures and with tissue explant cultures. The device is simply inserted into a standard 6-well plate, where it hangs self-supported at a distance of ~250 μm above the cell culture surface. The gradient is created by small microflows from the device, through an integrated track-etched porous membrane, into the cell culture well. The microfluidic transwell can deliver stable, quantifiable gradients over a large area with extremely low fluid shear stress to dissociated cells or tissue explants cultured independently on the surface of a 6-well plate. We used finite-element modeling to describe the porous membrane flow and molecular transport and to predict gradients generated by the device. Using the device, we applied a gradient of the chemotactic peptide N-Formyl-Met-Leu-Phe (fMLP) to a large population of HL-60 cells (a neutrophil cell line) and directly observed the migration with time-lapse microscopy. On quantification of the chemotactic response with an automated tracking algorithm, we found 74% of the cells moving towards the gradient. Additionally, the modular design and low fluid shear stress made it possible to apply gradients of growth factors and second messengers to mouse retinal explant cultures. With a simplified interface and well-defined gradients, the microfluidic transwell device has potential for broad applications to gradient-sensing biology. PMID:24225908

  16. Practical Bench Comparison of BBL CHROMagar Orientation and Standard Two-Plate Media for Urine Cultures

    PubMed Central

    D'Souza, Holly A.; Campbell, Mary; Baron, Ellen Jo

    2004-01-01

    A total of 1,023 urine samples sent for routine culture were plated onto sheep blood and MacConkey agars and a BBL CHROMagar Orientation (CO; Becton Dickinson, Cockeysville, Md.) plate, and the results were compared. Of these, 250 urine samples (24%) grew >10,000 CFU of one or two putative pathogens/ml and 773 showed no growth (NG), mixed growth of <10,000 CFU/ml, or three or more strains (mixed). The CO and conventional medium results agreed completely for 595 cultures with NG or <10,000 CFU/ml. An additional 178 urine samples yielded clinically insignificant differences. Both medium sets essentially agreed on quantities and identification for 400 single-pathogen cultures and 9 mixed cultures. With the caveat that CO cannot differentiate Klebsiella, Enterobacter, and Serratia spp., enteric pathogens were identified only by morphology and color on CO. Direct visual differentiation of group B streptococci from lactobacilli is not possible, but lactobacillus cells always exhibited easily recognizable morphology on Gram stain. Of 108 paired organism susceptibility results encompassing 2,268 drug-pathogen combinations, there were 3% errors and only 1% very major errors. Use of CO allowed a >50% reduction in inoculation time and a >20% reduction in work-up time. For our laboratory, with 50% “no growth” and ca. 25% significant results (50% Escherichia coli), CO allowed time and workup cost savings for a majority of cultures. A cost analysis (time and supplies for our laboratory) showed that if CO is used alone, the break-even level for CO pricing is $1.78; if CO and blood agar are both used, the break-even pricing for CO is $1.53. PMID:14715732

  17. Well plate-based perfusion culture device for tissue and tumor microenvironment replication.

    PubMed

    Zhang, W; Gu, Y; Hao, Y; Sun, Q; Konior, K; Wang, H; Zilberberg, J; Lee, W Y

    2015-07-01

    There are significant challenges in developing in vitro human tissue and tumor models that can be used to support new drug development and evaluate personalized therapeutics. The challenges include: (1) working with primary cells which are often difficult to maintain ex vivo, (2) mimicking native microenvironments from which primary cells are harvested, and (3) the lack of culture devices that can support these microenvironments to evaluate drug responses in a high-throughput manner. Here we report a versatile well plate-based perfusion culture device that was designed, fabricated and used to: (1) ascertain the role of perfusion in facilitating the expansion of human multiple myeloma cells and evaluate drug response of the cells, (2) preserve the physiological phenotype of primary murine osteocytes by reconstructing the 3D cellular network of osteocytes, and (3) circulate primary murine T cells through a layer of primary murine intestine epithelial cells to recapitulate the interaction of the immune cells with the epithelial cells. Through these diverse case studies, we demonstrate the device's design features to support: (1) the convenient and spatiotemporal placement of cells and biomaterials into the culture wells of the device; (2) the replication of tissues and tumor microenvironments using perfusion, stromal cells, and/or biomaterials; (3) the circulation of non-adherent cells through the culture chambers; and (4) conventional tissue and cell characterization by plate reading, histology, and flow cytometry. Future challenges are identified and discussed from the perspective of manufacturing the device and making its operation for routine and wide use. PMID:26021852

  18. Anti-epidermal-cell-surface pemphigus antibody detaches viable epidermal cells from culture plates by activation of proteinase.

    PubMed Central

    Farb, R M; Dykes, R; Lazarus, G S

    1978-01-01

    Immunoglobulin from pemphigus patients binds to the surface of mouse epidermal cells in culture. Cells incubated with the pemphigus antibody are easily detached from culture plates whereas cells incubated with serum from normal patients remain on the plate. Pemphigus antibody-mediated cell detachment is blocked by the addition of the proteinase inhibitors soybean trypsin inhibitor and alpha2-macroglobulin to the culture media. Detachable cells are viable, and activation of the complement cascade is not necessary for cell detachment. The anti-cell-surface antibody of pemphigus appears to disrupt adhesion between viable epidermal cells by activation of proteinase. Images PMID:272663

  19. Scanning electron microscopy of the nail plate in onychomycosis patients with negative fungal culture.

    PubMed

    Yue, Xueping; Li, Qing; Wang, Hongwei; Sun, Yilin; Wang, Aiping; Zhang, Qi; Zhang, Cuiping

    2016-03-01

    Onychomycosis is a common dermatological problem and can be identified by direct microscopic examination and fungal culture. However, the positive rate of fungal culture is low. This study investigated the application of scanning electron microscopy in the diagnosis of onychomycosis in 20 patients with negative fungal culture. In this study, a routine glutaraldehyde fixation method was used to prepare specimens for electron microscope examination. Results showed that under the scanning electron microscope, significant structural damage was observed in the nail plate in all patients. Hyphaes were seen in 70% of cases. A mixture of scattered hyphaes, pseudohyphaes, and spores was observed in 30% of cases. A mixture of spores and bacteria was observed in 10% of cases. A mixture of hyphaes and bacteria was observed in 20% of cases. The typical hyphae pierced a thin layer or single layer of corneocytes. Hyphaes could be smooth, sleek, and straight with visible separation, or dry, bent, and folded with a smooth surface. The diameter of hyphaes was 1-2 µm. The scattered spores were the main form of spore growth, and the growth of budding spores can be seen attached to the surface of layered armor. Most of the bacteria were gathered in clumps on the ventral surface, especially in grooves. In conclusion, scanning electron microscopy can be used to preliminarily identify the pathogen involved and the degree of damage in cases where onychomycosis is clinically diagnosed, but fungal culture is negative. SCANNING 38:172-176, 2016. © 2015 Wiley Periodicals, Inc. PMID:26291603

  20. Influence of plating density on individual cell growth, cell division and differentiation of neonatal rat heart primary cultures.

    PubMed

    Millart, H; Seraydarian, M W

    1986-01-01

    The influence of plating cell density of an originally enriched myocardial cell population has been studied in neonatal rat heart cells in culture. Low density (LDM) is defined as a density (24 h after plating) of 209 +/- 44 cells/mm2 (mean +/- SEM) and is compared with high density (HDM), 419 +/- 67 cells/mm2. Cell growth is evaluated by the total cell number, the percentage of myocardial cells (M) in culture (PAS method) and the protein content per cell. Some differentiation parameters such as beating rates, glycogen concentration, enzymatic activities (cytochrome C oxidase and glycogen phosphorylase) are studied with time in culture (48, 96 and 192 hr). High density was designed to yield a complete confluency of the cells within 24 hr after plating and to minimize cell division of the non-muscle cells (F). At high density, cell division of F cells is effectively limited, thus leading to a more stable model regarding the cell density per plate and the percentage of M cells: 85.7 +/- 4% and 33.4 +/- 6% in LDM cultures compared with 86.5 +/- 4.7% and 51.7 +/- 9.8% in HDM cultures at 24 and 192 hr (mean +/- SEM). Heart cells increase similarly in size with age in culture in both groups. In HDM cultures the spontaneous contractions begin sooner (24 hr) than in LDM cultures and are more rapidly synchronized. The beating rate is higher in HDM cultures between 48 and 96 hr; however, after this time it falls in HDM and does not fall in LDM. Thus the overgrowth of muscle cells by non-muscle cells is not responsible for loss of beating with time in culture but more likely high density could be a limiting factor for isotonic contraction. There is more glycogen per myocyte in LDM than in HDM cultures. The cell density influences the enzymatic activities of cytochrome C oxidase and glycogen phosphorylase. The cytochrome oxidase activity is higher in HDM cultures than in LDM cultures at 96 hr whereas glycogen phosphorylase activity is higher in LDM cultures at time 96 and 192

  1. How air influences radiation dose deposition in multiwell culture plates: a Monte Carlo simulation of radiation geometry

    PubMed Central

    Sabater, Sebastia; Berenguer, Roberto; Honrubia-Gomez, Paloma; Rivera, Miguel; Nuñez, Ana; Jimenez-Jimenez, Esther; Martos, Ana; Ramirez-Castillejo, Carmen

    2014-01-01

    Radiation of experimental culture cells on plates with various wells can cause a risk of underdosage as a result of the existence of multiple air–water interfaces. The objective of our study was to quantify this error in culture plates with multiple wells. Radiation conditions were simulated with the GAMOS code, based on the GEANT4 code, and this was compared with a simulation performed with PENELOPE and measured data. We observed a slight underdosage of ∼4% on the most superficial half of the culture medium. We believe that this underdosage does not have a significant effect on the dose received by culture cells deposited in a monolayer and adhered to the base of the wells. PMID:24722683

  2. Identification of Brucella by MALDI-TOF Mass Spectrometry. Fast and Reliable Identification from Agar Plates and Blood Cultures

    PubMed Central

    Ferreira, Laura; Vega Castaño, Silvia; Sánchez-Juanes, Fernando; González-Cabrero, Sandra; Menegotto, Fabiola; Orduña-Domingo, Antonio

    2010-01-01

    Background MALDI-TOF mass spectrometry (MS) is a reliable method for bacteria identification. Some databases used for this purpose lack reference profiles for Brucella species, which is still an important pathogen in wide areas around the world. We report the creation of profiles for MALDI-TOF Biotyper 2.0 database (Bruker Daltonics, Germany) and their usefulness for identifying brucellae from culture plates and blood cultures. Methodology/Principal Findings We created MALDI Biotyper 2.0 profiles for type strains belonging to B. melitensis biotypes 1, 2 and 3; B. abortus biotypes 1, 2, 5 and 9; B. suis, B. canis, B ceti and B. pinnipedialis. Then, 131 clinical isolates grown on plate cultures were used in triplicate to check identification. Identification at genus level was always correct, although in most cases the three replicates reported different identification at species level. Simulated blood cultures were performed with type strains belonging to the main human pathogenic species (B. melitensis, B. abortus, B. suis and B. canis), and studied by MALDI-TOF MS in triplicate. Identification at genus level was always correct. Conclusions/Significance MALDI-TOF MS is reliable for Brucella identification to the genus level from culture plates and directly from blood culture bottles. PMID:21151913

  3. The influence of the air plasma jet on early adherent events of L929 fibroblasts on cell culture polystyrene plate

    NASA Astrophysics Data System (ADS)

    Lee, Jung-Hwan; Kwon, Jae-Sung; Om, Ji-Yeon; Kim, Yong-Hee; Choi, Eun-Ha; Kim, Kwang-Mahn; Kim, Kyoung-Nam

    2012-10-01

    Recently, atmospheric pressure plasma was applied to biological field. The aim of this study was to identify whether the air plasma jet increases fibroblast early attachment under moving motion on the cell culture polystyrene plate. Polystyrene plate was treated with plasma jet using compressed air. After 2 minutes of treatment, L929 was seeded on polystyrene plate as well as on untreated plate. Cells were allowed to attach for 4 hours under 70 RPM. FE-SEM, confocal microscopy and RT-PCR were used to evaluate characters of cells. The results suggested that plasma treatment on the polystyrene plate altered surface energy without change of roughness. In occasion of treatment plate, attached L292 were significantly found but not found on untreated surface. Also, despite the small area of treated center by the flame of the plasma jet, cells were also attached on round surface of the area covered by the flame, which suggests that the effect was not only due to the jet flame but perhaps due to the jet interacting with surrounding atmosphere. In the light of this study, the air plasma jet could be useful for early attachment of L292 on the polystyrene plate under moving motion and can be applied to biomaterials.

  4. Fluorometric assessment of acetaminophen-induced toxicity in rat hepatocyte spheroids seeded on micro-space cell culture plates.

    PubMed

    Sanoh, Seigo; Santoh, Masataka; Takagi, Masashi; Kanayama, Tatsuya; Sugihara, Kazumi; Kotake, Yaichiro; Ejiri, Yoko; Horie, Toru; Kitamura, Shigeyuki; Ohta, Shigeru

    2014-09-01

    Hepatotoxicity induced by the metabolic activation of drugs is a major concern in drug discovery and development. Three-dimensional (3-D) cultures of hepatocyte spheroids may be superior to monolayer cultures for evaluating drug metabolism and toxicity because hepatocytes in spheroids maintain the expression of various metabolizing enzymes and transporters, such as cytochrome P450 (CYP). In this study, we examined the hepatotoxicity due to metabolic activation of acetaminophen (APAP) using fluorescent indicators of cell viability and intracellular levels of glutathione (GSH) in rat hepatocyte spheroids grown on micro-space cell culture plates. The mRNA expression levels of some drug-metabolizing enzymes were maintained during culture. Additionally, this culture system was compatible with microfluorometric imaging under confocal laser scanning microscopy. APAP induced a decrease in intracellular ATP at 10mM, which was blocked by the CYP inhibitor 1-aminobenzotriazole (ABT). APAP (10mM, 24h) decreased the levels of both intracellular ATP and GSH, and GSH-conjugated APAP (APAP-GSH) were formed. All three effects were blocked by ABT, confirming a contribution of APAP metabolic activation by CYP to spheroid toxicity. Fluorometric imaging of hepatocyte spheroids on micro-space cell culture plates may allow the screening of drug-induced hepatotoxicity during pharmaceutical development. PMID:24878114

  5. Effect of nitrogen-rich cell culture surfaces on type X collagen expression by bovine growth plate chondrocytes

    PubMed Central

    2011-01-01

    Background Recent evidence indicates that osteoarthritis (OA) may be a systemic disease since mesenchymal stem cells (MSCs) from OA patients express type X collagen, a marker of late stage chondrocyte hypertrophy (associated with endochondral ossification). We recently showed that the expression of type X collagen was suppressed when MSCs from OA patients were cultured on nitrogen (N)-rich plasma polymer layers, which we call "PPE:N" (N-doped plasma-polymerized ethylene, containing up to 36 atomic percentage (at.% ) of N. Methods In the present study, we examined the expression of type X collagen in fetal bovine growth plate chondrocytes (containing hypertrophic chondrocytes) cultured on PPE:N. We also studied the effect of PPE:N on the expression of matrix molecules such as type II collagen and aggrecan, as well as on proteases (matrix metalloproteinase-13 (MMP-13) and molecules implicated in cell division (cyclin B2). Two other culture surfaces, "hydrophilic" polystyrene (PS, regular culture dishes) and nitrogen-containing cation polystyrene (Primaria®), were also investigated for comparison. Results Results showed that type X collagen mRNA levels were suppressed when cultured for 4 days on PPE:N, suggesting that type X collagen is regulated similarly in hypertrophic chondrocytes and in human MSCs from OA patients. However, the levels of type X collagen mRNA almost returned to control value after 20 days in culture on these surfaces. Culture on the various surfaces had no significant effects on type II collagen, aggrecan, MMP-13, and cyclin B2 mRNA levels. Conclusion Hypertrophy is diminished by culturing growth plate chondrocytes on nitrogen-rich surfaces, a mechanism that is beneficial for MSC chondrogenesis. Furthermore, one major advantage of such "intelligent surfaces" over recombinant growth factors for tissue engineering and cartilage repair is potentially large cost-saving. PMID:21244651

  6. Electrical excitability of outgrowing neurites of embryonic neurones in cultures of dissociated neural plate of Xenopus laevis.

    PubMed Central

    Willard, A L

    1980-01-01

    1. I have studied the electrical excitability of outgrowing processes of individual neurones in cultures made from dissociated neural plates of embryos of Xenopus laevis prior to the time of neurite outgrowth in vivo. 2. The electrical excitability of neurites was tested by stimulating them extracellularly and recording responses with an intracellular electrode in their cell bodies; neurites were excitable at all times examined. 3. The ionic basis of the excitability of neurites was tested by recording from cells while changing the composition of the salines perfusing the cultures. 4. In cultures less than 10 hr old, all neurites tested made responses which depended on Ca2+. The action potentials of the cell bodies were also Ca2+-dependent at these times. 5. Between 10 and 12 hr in culture, a time at which the cell bodies still made Ca2+-dependent action potentials, neurites acquired the ability to make Na+-dependent responses. At these times, two-thirds of neurites tested retained the ability to produce divalent cation-dependent action potentials when perfused with solutions of isotonic Ba2+. 6. After 12 hr in culture, no neurites were observed to make Ca2+-or Ba2+-dependent responses; only Na+-dependent responses were observed. Cells continued to initiate and elongate new neurites until about 24 hr in culture. Thus neurites sent out at different times in culture differed in their development of excitability. 7. Cell bodies making exclusively Ca2+-dependent action potentials could be found until about 15 hr in culture, after which time a Na+-dependent component appeared. Cell bodies could then be observed to make action potentials which depended on both Ca2+ and Na+ until about 3 days in culture. After 3 days, most cell bodies made predominately Na+-dependent action potentials. Unlike the neurites, cell bodies retained the ability to make action potentials in isotonic Ba2+ for as long as the cultures were maintained (up to 5 days). 8. The possibility that changes

  7. Soluble penicillin-binding protein 2a: beta-lactam binding and inhibition by non-beta-lactams using a 96-well format.

    PubMed

    Toney, J H; Hammond, G G; Leiting, B; Pryor, K D; Wu, J K; Cuca, G C; Pompliano, D L

    1998-01-01

    High level methicillin resistance in Staphylococcus aureus is dependent upon the acquisition of the mecA gene encoding penicillin-binding protein 2a (PBP2a). PBP2a is a member of a family of peptidoglycan biosynthetic enzymes involved in assembly of the cell wall in bacteria and is poorly inactivated by beta-lactam antibiotics. We describe a 96-well-filter binding assay using recombinant, soluble PBP2a which allows for kinetic measurement of penicillin binding. The deacylation rate constant for the PBP2a-penicillin G covalent complex was found to be 5.7 +/- 1.0 x 10(-5) s-1 at 30 degrees C (half-life of approximately 200 min). For the PBP2a acylation reaction, the value of K(m) (penicillin G) = 0.5 +/- 0.1 mM and kcat = 1 x 10(-3) s-1, which yields a second-order rate constant (kcat/K(m)) for inactivation of 2.0 M-1 s-1. Using this assay, several non-beta-lactam inhibitors including Cibacron blue have been found which exhibit IC50 values between 10 and 30 microM. The binding affinities of several carbapenems and beta-lactams correlated well between the filter binding assay described in this report and an electrophoretic assay for PBP2a using membranes prepared form methicillin-resistant S. aureus. PMID:9448849

  8. Paper microzone plates.

    PubMed

    Carrilho, Emanuel; Phillips, Scott T; Vella, Sarah J; Martinez, Andres W; Whitesides, George M

    2009-08-01

    This paper describes 96- and 384-microzone plates fabricated in paper as alternatives to conventional multiwell plates fabricated in molded polymers. Paper-based plates are functionally related to plastic well plates, but they offer new capabilities. For example, paper-microzone plates are thin (approximately 180 microm), require small volumes of sample (5 microL per zone), and can be manufactured from inexpensive materials ($0.05 per plate). The paper-based plates are fabricated by patterning sheets of paper, using photolithography, into hydrophilic zones surrounded by hydrophobic polymeric barriers. This photolithography used an inexpensive formulation photoresist that allows rapid (approximately 15 min) prototyping of paper-based plates. These plates are compatible with conventional microplate readers for quantitative absorbance and fluorescence measurements. The limit of detection per zone loaded for fluorescence was 125 fmol for fluorescein isothiocyanate-labeled bovine serum albumin, and this level corresponds to 0.02 the quantity of analyte per well used to achieve comparable signal-to-noise in a 96-well plastic plate (using a solution of 25 nM labeled protein). The limits of detection for absorbance on paper was approximately 50 pmol per zone for both Coomassie Brilliant Blue and Amaranth dyes; these values were 0.4 that required for the plastic plate. Demonstration of quantitative colorimetric correlations using a scanner or camera to image the zones and to measure the intensity of color, makes it possible to conduct assays without a microplate reader. PMID:19572563

  9. Effects of parathyroid hormone and calcitonin on alkaline phosphatase activity and matrix calcification in rabbit growth-plate chondrocyte cultures

    SciTech Connect

    Kato, Y.; Shimazu, A.; Nakashima, K.; Suzuki, F.; Jikko, A.; Iwamoto, M. )

    1990-07-01

    The effects of PTH and calcitonin (CT) on the expression of mineralization-related phenotypes by chondrocytes were examined. In cultures of pelleted growth-plate chondrocytes. PTH caused 60-90% decreases in alkaline phosphatase activity, the incorporation of {sup 45}Ca into insoluble material, and the calcium content during the post-mitotic stage. These effects of PTH were dose-dependent and reversible. In contrast, CT increased alkaline phosphatase activity, {sup 45}Ca incorporation into insoluble material, and the calcium content by 1.4- to 1.8-fold. These observations suggest that PTH directly inhibits the expression of the mineralization-related phenotypes by growth-plate chondrocytes, and that CT has the opposite effects.

  10. Determination of the HIV integrase inhibitor, MK-0518 (raltegravir), in human plasma using 96-well liquid-liquid extraction and HPLC-MS/MS.

    PubMed

    Merschman, S A; Vallano, P T; Wenning, L A; Matuszewski, B K; Woolf, E J

    2007-09-15

    An HPLC-MS/MS method was developed for the determination of MK-0518 (raltegravir), an HIV integrase inhibitor, in human plasma over the concentration range of 2-1000 ng/mL. Stable isotope labeled (13)C(6)-MK-0518 was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction with hexane:methylene chloride in the 96-well format with a 200 microL plasma sample size. The compounds were chromatographed on an Ace C(18) (50 x 3.0 mm, 3 microm, titanium frits) column with 42.5/57.5 (v/v %) 0.1mM EDTA in 0.1% formic acid/methanol mobile phase at a flow rate of 0.5 mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for MK-0518 (m/z 445-->109) and (13)C(6)-MK-0518 (m/z 451-->367) on an Applied Biosystem API 4000 HPLC-MS/MS was used for quantitation. Intraday precision of standard curve concentrations in five different lots of control plasma was within 3.2%, while accuracy ranged from 94.8 to 106.8%. The mean extraction recovery of spiked plasma samples was 87%. Quality control (QC) samples were stored at -20 degrees C. Initial within day analysis showed QC accuracy within 7.5% of nominal with precision of 3.1% or less. The plasma QC samples were demonstrated to be stable for up to 23 months at -20 degrees C. The method described has been used to support over 18 clinical studies during Phase I through III of clinical development. PMID:17644453

  11. Trace Amounts of Furan-2-Carboxylic Acids Determine the Quality of Solid Agar Plates for Bacterial Culture

    PubMed Central

    Hara, Shintaro; Isoda, Reika; Tahvanainen, Teemu; Hashidoko, Yasuyuki

    2012-01-01

    Background Many investigators have recognised that a significant proportion of environmental bacteria exist in a viable but non-culturable state on agar plates, and some researchers have also noticed that some of such bacteria clearly recover their growth on matrices other than agar. However, the reason why agar is unsuitable for the growth of some bacteria has not been addressed. Methodology/Principal Findings According to the guide of a bioassay for swarming inhibition, we identified 5-hydroxymethylfuran-2-carboxylic acid (5-HMFA) and furan-2-carboxylic acid (FA) as factors that inhibit bacterial swarming and likely inhibit extracellular polysaccharide production on agar. The furan-2-carboxylic acids 5-HMFA and FA effectively inhibited the swarming and swimming of several environmental bacteria at concentrations of 1.8 and 2.3 µg L−1 (13 and 21 nmol L−1), respectively, which are equivalent to the concentrations of these compounds in 0.3% agar. On Luria-Bertani (LB) plates containing 1.0% agar that had been previously washed with MeOH, a mixture of 5-HMFA and FA in amounts equivalent to their original concentrations in the unwashed agar repressed the swarming of Escherichia coli K12 strain W3110, a representative swarming bacterium. Conclusions/Significance Agar that contains trace amounts of 5-HMFA and FA inhibits the proliferation of some slow-growing or difficult-to-culture bacteria on the plates, but it is useful for single colony isolation due to the ease of identification of swarmable bacteria as the non-swarmed colonies. PMID:22848437

  12. A review of the current state of digital plate reading of cultures in clinical microbiology.

    PubMed

    Rhoads, Daniel D; Novak, Susan M; Pantanowitz, Liron

    2015-01-01

    Digital plate reading (DPR) is increasingly being adopted as a means to facilitate the analysis and improve the quality and efficiency within the clinical microbiology laboratory. This review discusses the role of DPR in the context of total laboratory automation and explores some of the platforms currently available or in development for digital image capturing of microbial growth on media. The review focuses on the advantages and challenges of DPR. Peer-reviewed studies describing the utility and quality of these novel DPR systems are largely lacking, and professional guidelines for DPR implementation and quality management are needed. Further development and more widespread adoption of DPR is anticipated. PMID:26110091

  13. A review of the current state of digital plate reading of cultures in clinical microbiology

    PubMed Central

    Rhoads, Daniel D.; Novak, Susan M.; Pantanowitz, Liron

    2015-01-01

    Digital plate reading (DPR) is increasingly being adopted as a means to facilitate the analysis and improve the quality and efficiency within the clinical microbiology laboratory. This review discusses the role of DPR in the context of total laboratory automation and explores some of the platforms currently available or in development for digital image capturing of microbial growth on media. The review focuses on the advantages and challenges of DPR. Peer-reviewed studies describing the utility and quality of these novel DPR systems are largely lacking, and professional guidelines for DPR implementation and quality management are needed. Further development and more widespread adoption of DPR is anticipated. PMID:26110091

  14. Optimization of in situ hybridization for detection of viral genomes in cultured cells on 96-microwell plates: a cytomegalovirus model.

    PubMed Central

    Mougin, C; Bassignot, A; Coaquette, A; Bourgeois, A; Lab, M

    1991-01-01

    In situ hybridization (ISH) for identification of infectious replicative cytomegalovirus (CMV) in cell culture microplates (96 microwells) infected by clinical specimens was tested by using a biotin-labeled DNA probe and an avidin-alkaline phosphatase conjugate. A total of 395 specimens were examined by using ISH and a monoclonal antibody (MAb) specific for an early antigen of CMV. Of 47 specimens that gave a positive signal for CMV by ISH, 33 were confirmed virus positive by MAb staining. Of 141 blood samples tested, 4.96% were positive by ISH, and 0.7% were positive by the MAb technique. ISH shows 40% more sensitivity than MAb staining. This technique should be widely applicable for the specific identification of viral isolates (e.g., herpesvirus, myxovirus, paramyxovirus, and enterovirus) in cell culture 96-microwell microplates, thereby making it feasible to screen a larger number of samples than is possible with classical methods using conventional culture tubes, shell vials, or 24-well plates. Images PMID:1662228

  15. Simultaneous determination of a novel KDR kinase inhibitor and its N-oxide metabolite in human plasma using 96-well solid-phase extraction and liquid chromatography/tandem mass spectrometry.

    PubMed

    Xu, Yang; Du, Lihong; Soli, Eric D; Braun, Matthew P; Dean, Dennis C; Musson, Donald G

    2005-03-25

    To support pharmacokinetic studies, a selective and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous determination of a novel KDR kinase inhibitor (1) and its active metabolite (2) in human plasma. The method is fully automated using a Packard MultiPROBE II system and a TomTec Quadra 96 liquid handling workstation to perform sample preparation and solid-phase extraction (SPE). Following the extraction on a mixed-mode SPE using Oasis MCX 96-well plate, the analytes were separated on a Aquasil C18 column (50 mm x 2.1 mm, i.d., 3 microm) with a mobile phase consisting of acetonitrile/ammonium acetate buffer (5 mM, pH 5.0) (60/40, v/v). The run time for each injection was 4.5 min with the retention times of approximately 2.0 and 2.7 min for 1 and 2 respectively, at a flow rate of 0.25 mL/min. A tandem mass spectrometric detection was conducted using multiple reaction monitoring (MRM) under the positive ion mode with a turbo ion-spray interface. The linear ranges of the calibration curves were 0.05-400 ng/mL for 1 and 0.1-400 ng/mL for 2 on a PE Sciex API 4000 LC-MS/MS system. The lower limits of quantitation (LLOQ) of the assay were 0.05 and 0.1 ng/mL for 1 and 2 respectively, when 0.4 mL of plasma was processed. Intra-day assay precision (using five standard curves prepared by spiking compounds to five lots of plasma) was less than 4.9% for 1 and less than 9.6% for 2 on each concentration. Assay accuracy was found to be 95.1-104.6% of nominal for 1 standards and 93.5-105.6% for 2 standards. QC samples were stable when kept at room temperature for 4 h, at -70 degrees C for 10 days, and after three freeze-thaw cycles. The extraction recoveries were 80%, 83% and 84% for 1 and 2 and I.S. respectively, and no significant matrix effects were observed. The method was successfully applied to plasma samples from clinical studies after oral administration of compound 1. PMID:15686997

  16. Comparison of diploid fibroblast and rabbit kidney tissue cultures and a diploid fibroblast microtiter plate system for the isolation of herpes simplex virus.

    PubMed Central

    Langenberg, A; Zbanyszek, R; Dragavon, J; Ashley, R; Corey, L

    1988-01-01

    We evaluated the relative sensitivities of two cell systems (rabbit kidney [RK] and human diploid fibroblast [DF; human embryonic tonsil]) in standard tube cultures versus DF cells in a 48-well microtiter plate system for the detection of both symptomatic and asymptomatic herpes simplex virus (HSV) infection. At least one system isolated HSV in 111 of 809 specimens (13.7%). HSV was isolated in RK tube cultures from 110 specimens (99%), in DF tube cultures from 91 specimens (82%), and in DF microtiter plates from 95 specimens (86%). The frequency of HSV isolation varied with the anatomic site and the presence or absence of a herpetic lesion. The sensitivities of the three culture systems remained similar whether the specimens were obtained from lesions or whether the specimens were taken to determine if asymptomatic excretion of HSV was present. While RK tube cultures were more sensitive than DF tube cultures, the DF microtiter plate system was as sensitive as DF tube cultures and its use is supported as a cheaper and less labor-intensive method for the detection of HSV. PMID:2846647

  17. Biodiesel production from marine cyanobacteria cultured in plate and tubular photobioreactors.

    PubMed

    Selvan, B Karpanai; Revathi, M; Piriya, P Sobana; Vasan, P Thirumalai; Prabhu, D Immuanual Gilwax; Vennison, S John

    2013-03-01

    Carbon (neutral) based renewable liquid biofuels are alternative to petroleum derived transport fuels that contribute to global warming and are of a limited availability. Microalgae based biofuels are considered as promising source of energy. Lyngbya sp. and Synechococcus sp. were studied for the possibility of biodiesel production in different media such as ASNIII, sea water enrichment medium and BG11. The sea water enrichment medium was found superior in enhancing the growth rate of these microalgae. Nitrogen depletion has less effect in total chlorophyll a content, at the same time the lipid content was increased in both Lyngbya sp. and Synechococcus sp. by 1.4 and 1.2 % respectively. Increase in salinity from 0.5-1.0 M also showed an increase in the lipid content to 2.0 and 0.8 % in these strains; but a salinity of 1.5 M has a total inhibitory effect in the growth. The total biomass yield was comparatively higher in tubular LED photobioreactor than the fluorescent flat plated photobioreactor. Lipid extraction was obtained maximum at 60 degrees C in 1:10 sample: solvent ratio. GC-MS analysis of biodiesel showed high content of polyunsaturated fatty acids (PUFA; 4.86 %) than saturated fatty acid (SFA; 4.10 %). Biodiesel production was found maximum in Synechococcus sp. than Lyngbya sp. The viscosity of the biodiesel was closely related to conventional diesel. The results strongly suggest that marine microalgae could be used as a renewable energy source for biodiesel production. PMID:23678548

  18. Culture of Isolated Floor Plate Tissue and Production of Conditioned Medium to Assess Functional Properties of Floor Plate-released Signals

    PubMed Central

    Charoy, Camille; Arbeille, Elise; Thoinet, Karine; Castellani, Valérie

    2014-01-01

    During development, progenitors and post-mitotic neurons receive signals from adjacent territories that regulate their fate. The floor-plate is a group of glial cells lining the ependymal canal at ventral position. The floor-plate expresses key morphogens contributing to the patterning of cell lineages in the spinal cord. At later developmental stages, the floor-plate regulates the navigation of axons in the spinal cord, acting as a barrier to prevent the crossing of ipsilateral axons and controlling midline crossing by commissural axons1. These functions are achieved through the secretion of various guidance cues. Some of these cues act as attractants and repellents for the growing axons while others regulate guidance receptors and downstream signaling to modulate the sensitivity of the axons to the local guidance cues2,3. Here we describe a method that allows investigating the properties of floor-plate derived signals in a variety of developmental contexts, based on the production of Floor-Plate conditioned medium (FPcm)4-6. We then exemplify the use of this FPcm in the context of axon guidance. First, the spinal cord is isolated from mouse embryo at E12.5 and the floor-plate is dissected out and cultivated in a plasma-thrombin matrix (Figure 1). Second two days later, commissural tissue are dissected out from E12.5 embryos, triturated and exposed to the FPcm. Third, the tissue are processed for Western blot analysis of commissural markers. PMID:24561889

  19. Effect of different heterotrophic plate count methods on the estimation of the composition of the culturable microbial community

    PubMed Central

    Gössl, Eva-Maria; Antonielli, Livio; Sessitsch, Angela; Kostić, Tanja

    2015-01-01

    Heterotrophic plate counts (HPC) are routinely determined within the scope of water quality assessment. However, variable HPC methods with different cultivation parameters (i.e., temperature and media type) are applied, which could lead to significant effects in the outcome of the analysis. Therefore the effect of different HPC methods, according to DIN EN ISO 6222 and EPA, on the culturable microbial community composition was investigated by 16S rRNA gene sequence analysis and statistical evaluation was performed. The culturable community composition revealed significant effects assigned to temperature (p < 0.01), while for media type no statistical significance was observed. However, the abundance of certain detected bacteria was affected. Lower temperature (22 °C) showed the abundance of naturally occurring Pseudomonadaceae and Aeromonadaceae, whereas at high temperature (37 °C) numerous Enterobacteriaceae, Citrobacter spp. and Bacilli were identified. The highest biodiversity was detected at lower temperature, especially on R2A medium. These results indicate that different temperatures (low and high) should be included into HPC measurement and selection of media should, ideally, be adjusted to the monitored water source. Accordingly, it can be inferred that the HPC method is more suitable for continuous monitoring of the same water source than for single assessments of a water sample. PMID:25861554

  20. Comparison of the TEMPO® System, Petrifilm® , and Cultural MPN Procedure for Enumeration of E. coli, Coliforms and Total Aerobic Plate Counts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Recent innovations in microbiological methods for analysis of food products have been in methods for detection of bacterial pathogens. Petrifilm dehydrated plates are the only significant addition to cultural procedures for indicator organisms in the last 20 years. An automated most...

  1. Hyperspectral imaging for detection of non-O157 shiga-toxin producing escherichia coli(STEC) serogroups on spread plates of mixed cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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. Althou...

  2. A comparison of standard cultural methods for the detection of foodborne Salmonella species including three new chromogenic plating media.

    PubMed

    Schönenbrücher, Vanessa; Mallinson, Edward T; Bülte, Michael

    2008-03-31

    In this study the draft of the horizontal method for the detection of Salmonella species from human food and animal feed (ISO 6579:2002) was compared to the European gold standard (DIN EN 12824:1998), including the three new chromogenic plating media AES Salmonella Agar Plate (ASAP), Oxoid Salmonella Chromogen Media (OSCM) and Miller-Mallinson agar (MM). First the growth and appearance of 36 bacterial type strains (Salmonella and other 21 species) on ASAP, OSCM and MM were compared to those on the three traditional agars Brilliant Green Agar according to Edel and Kampelmacher (BGA), Xylose Lysine Deoxycholate Agar (XLD) and Xylose Lysine Tergitol 4 Agar (XLT4). Only on MM agar, did all of 36 tested type strains produce typical colonies, especially strains of S. Senftenberg, Salmonella arizonae, S. Dublin and S. Derby. Artificial inoculation experiments using raw pork ground meat (n=92) were subsequently conducted. A shortened incubation time of 24 h in RVS broth yielded a Salmonella species recovery of 100% from spiked meat samples. Finally, 286 naturally contaminated raw porcine and bovine minced meat samples and raw poultry meat samples were investigated. Forty-three strains from a total of 39 Salmonella-positive samples were found. S. Typhimurium (n=21), with DT 104 L, DT 012 and RDNC being the most prevalent subtypes isolated. D-tartrate-positive S. Paratyphi B (n=2) and S. Saint-Paul (n=3) were also recovered. They were cultured from poultry meat and were multi-resistant against antibiotics including nalidixic acid. Rappaport Vassiliadis broth with soypeptone (RVS) yielded the highest recovery of Salmonella spp. (97,4%) compared to Tetrathionate broth with Novobiocin according to Muller and Kauffman (MKTTn, 94,9%) and Selenite Cystine broth (SC, 38,5%). However, no significant difference was obtained by comparing the ISO 6579:2002 draft to the gold standard. PMID:18192050

  3. Mechanical influence of tissue culture plates and extracellular matrix on mesenchymal stem cell behavior: A topical review.

    PubMed

    Tatullo, Marco; Marrelli, Massimo; Falisi, Giovanni; Rastelli, Claudio; Palmieri, Francesca; Gargari, Marco; Zavan, Barbara; Paduano, Francesco; Benagiano, Vincenzo

    2016-03-01

    Tissue engineering applications need a continuous development of new biomaterials able to generate an ideal cell-extracellular matrix interaction. The stem cell fate is regulated by several factors, such as growth factors or transcription factors. The most recent literature has reported several publications able to demonstrate that environmental factors also contribute to the regulation of stem cell behavior, leading to the opinion that the environment plays the major role in the cell differentiation.The interaction between mesenchymal stem cells (MSCs) and extracellular environment has been widely described, and it has a crucial role in regulating the cell phenotype. In our laboratory (Tecnologica Research Institute, Crotone, Italy), we have recently studied how several physical factors influence the distribution and the morphology of MSCs isolated from dental pulp, and how they are able to regulate stem cell differentiation. Mechanical and geometrical factors are only a small part of the environmental factors able to influence stem cell behavior, however, this influence should be properly known: in fact, this assumption must be clearly considered during those studies involving MSCs; furthermore, these interactions should be considered as an important bias that involves an high number of studies on the MSCs, since in worldwide laboratories the scientists mostly use tissue culture plates for their experiments. PMID:26612837

  4. The gene expression pattern induced by high plating density in cultured bovine and buffalo granulosa cells might be regulated by specific miRNA species

    PubMed Central

    YENUGANTI, Vengala Rao; BADDELA, Vijay Simha; BAUFELD, Anja; SINGH, Dheer; VANSELOW, Jens

    2015-01-01

    Precise regulation of cell type-specific gene expression profiles precedes the profound morphological reorganization of somatic cell layers during folliculogenesis, ovulation and luteinization. Cell culture models are essential to the study of corresponding molecular mechanisms of gene regulation. In a recent study, it was shown that an increased cell plating density can largely change gene expression profiles of cultured bovine granulosa cells. In our present study, we comparatively analyzed cell plating density effects on cultured bovine and buffalo granulosa cells. Cells were isolated from small- to medium-sized follicles (2–6 mm) and cultured under serum-free conditions at different plating densities. The abundance of selected marker transcripts and associated miRNA candidates was determined by quantitative real-time RT-PCR. We found in both species that the abundance of CYP19A1, CCNE1 and PCNA transcripts was remarkably lower at a high plating density, whereas VNN2 and RGS2 transcripts significantly increased. In contrast, putative regulators of CYP19A1, miR-378, miR-106a and let-7f were significantly higher in both species or only in buffalo, respectively. Also miR-15a, a regulator of CCNE1, was upregulated in both species. Thus, increased plating density induced similar changes of mRNA and miRNA expression in granulosa cells from buffalo and cattle. From these data, we conclude that specific miRNA species might be involved in the observed density-induced gene regulation. PMID:25740097

  5. Culturing in serum-free culture medium on collagen type-I-coated plate increases expression of CD133 and retains original phenotype of HT-29 cancer stem cell

    PubMed Central

    Arab-Bafrani, Zahra; Shahbazi-Gahrouei, Daryoush; Abbasian, Mehdi; Saberi, Alihossein; Fesharaki, Mehrafarin; Hejazi, Seyed Hossein; Manshaee, Samira

    2016-01-01

    Background: A sub-population of tumor cells termed cancer stem cells (CSCs) has an important role in tumor initiation, progression, and recurrence. Selecting a suitable procedure for isolation and enrichment of CSCs is the biggest challenge in the study of CSCs. In the present study, the role of the combination of stem cell culture medium and collagen type-I was evaluated for successful isolation and enrichment of HT-29 CSCs. Materials and Methods: HT-29 cells were cultured in serum-containing medium (parental culture medium: Medium + 10% fetal bovine serum) and serum-free medium (stem cell culture medium); both on collagen-coated plates. Spheres forming ability and CD133 expression, as a potential marker of colorectal CSCs, were evaluated in two culture mediums. Results: The results show spheroids usually give rise completely within 15 days in the stem cell culture medium on the collagen-coated plate. CD133 expression in spheroid cells (84%) is extensively higher than in parental cells (25%). Moreover, relative to parental cells, spheroid cells were more radioresistance. Conclusion: Finding of this study suggested that CSCs derived from colon cancer cell line (HT-29) can be propagated and form colonospheres in serum-free culture medium on collagen type-I. According to maintenance of their original phenotype in these conditions, it seems serum-free culture medium on collagen type-I is a suitable way to drug screening of HT-29 CSCs. PMID:27135028

  6. Enumeration of total coliforms and E. coli in foods by the SimPlate coliform and E. coli color indicator method and conventional culture methods: collaborative study.

    PubMed

    Feldsine, Philip T; Lienau, Andrew H; Roa, Nerie H; Green, Shannon T

    2005-01-01

    The relative effectiveness of the SimPlate Coliform and E. coli Color Indicator (CEc-CI) method was compared to the AOAC 3-tube Most Probable Number (MPN) methods for enumerating and confirming coliforms and Escherichia coli in foods (966.23 and 966.24). In this study, test portions were prepared and analyzed according to the conditions stated in both the AOAC methods and SimPlate directions for use. Six food types were artificially contaminated with coliform bacteria and E. coli: frozen burritos, frozen broccoli, fluid pasteurized milk, whole almond nut meats, cheese, and powdered cake mix. Method comparisons were conducted. Overall, the SimPlate method demonstrated <0.3 log difference for total coliform and E. coli counts compared to the AOAC reference methods for the majority of food types and levels analyzed. In all cases, the repeatability and reproducibility of the SimPlate CEc-CI method were not different from those of the reference methods and in certain cases, were statistically better than those of the AOAC 3-tube MPN methods. These results indicate that the SimPlate CEc-CI method and the reference culture methods are comparable for enumeration of both total coliforms and E. coli in foods. PMID:16385981

  7. 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

  8. Comparison of polymerase chain reaction methods and plating for analysis of enriched cultures of Listeria monocytogenes when using the ISO11290-1 method.

    PubMed

    Dalmasso, Marion; Bolocan, Andrei Sorin; Hernandez, Marta; Kapetanakou, Anastasia E; Kuchta, Tomáš; Manios, Stavros G; Melero, Beatriz; Minarovičová, Jana; Muhterem, Meryem; Nicolau, Anca Ioana; Rovira, Jordi; Skandamis, Panagiotis N; Stessl, Beatrix; Wagner, Martin; Jordan, Kieran; Rodríguez-Lázaro, David

    2014-03-01

    Analysis for Listeria monocytogenes by ISO11290-1 is time-consuming, entailing two enrichment steps and subsequent plating on agar plates, taking five days without isolate confirmation. The aim of this study was to determine if a polymerase chain reaction (PCR) assay could be used for analysis of the first and second enrichment broths, saving four or two days, respectively. In a comprehensive approach involving six European laboratories, PCR and traditional plating of both enrichment broths from the ISO11290-1 method were compared for the detection of L. monocytogenes in 872 food, raw material and processing environment samples from 13 different dairy and meat food chains. After the first and second enrichments, total DNA was extracted from the enriched cultures and analysed for the presence of L. monocytogenes DNA by PCR. DNA extraction by chaotropic solid-phase extraction (spin column-based silica) combined with real-time PCR (RTi-PCR) was required as it was shown that crude DNA extraction applying sonication lysis and boiling followed by traditional gel-based PCR resulted in fewer positive results than plating. The RTi-PCR results were compared to plating, as defined by the ISO11290-1 method. For first and second enrichments, 90% of the samples gave the same results by RTi-PCR and plating, whatever the RTi-PCR method used. For the samples that gave different results, plating was significantly more accurate for detection of positive samples than RTi-PCR from the first enrichment, but RTi-PCR detected a greater number of positive samples than plating from the second enrichment, regardless of the RTi-PCR method used. RTi-PCR was more accurate for non-food contact surface and food contact surface samples than for food and raw material samples especially from the first enrichment, probably because of sample matrix interference. Even though RTi-PCR analysis of the first enrichment showed less positive results than plating, in outbreak scenarios where a rapid result is

  9. β-Cyclodextrin enhanced on-line organic solvent field-amplified sample stacking in capillary zone electrophoresis for analysis of ambroxol in human plasma, following liquid-liquid extraction in the 96-well format.

    PubMed

    Li, Ji; Bi, Youwei; Wang, Li; Sun, Fanlu; Chen, Zhao; Xu, Guili; Fan, Guorong

    2012-07-01

    A field-amplified sample stacking (FASS) and capillary zone electrophoresis (CZE) method is described for the quantification of ambroxol hydrochloride in human plasma, following liquid-liquid extraction in the 96-well format. The separation was carried out at 25 °C in a 31.2 cm × 75 μm fused-silica capillary with an applied voltage of 15 kV. The background electrolyte (BGE) was composed of 6.25 mM borate-25 mM phosphate (pH 3.0) and 1mM β-cyclodextrin. The detection wavelength was 210 nm. Clean-up and preconcentration of plasma biosamples were developed by 96-well format liquid-liquid extraction (LLE). In this study, FASS in combination with β-cyclodextrin enhanced the sensitivity about 60-70 fold in total. The method was suitably validated with respect to stability, specificity, linearity, lower limit of quantitation, accuracy, precision, extraction recovery and robustness. The calibration graph was linear for ambroxol hydrochloride from 2 to 500 ng/ml. The lower limit of quantification was 2 ng/ml. The intra- and inter-day precisions of lowest limit of quantification (LLOQ) were 9.61 and 11.80%, respectively. The method developed was successfully applied to the evaluation of clinical pharmacokinetic study of ambroxol hydrochloride tablet after oral administration to 12 healthy volunteers. PMID:22464560

  10. Microtiter plate-based antibody microarrays for bacteria and toxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Research has focused on the development of rapid biosensor-based, high-throughput, and multiplexed detection of pathogenic bacteria in foods. Specifically, antibody microarrays in 96-well microtiter plates have been generated for the purpose of selective detection of Shiga toxin-producing E. coli (...

  11. LED illumination effects on proliferation and survival of meningioma cellular cultures

    NASA Astrophysics Data System (ADS)

    Solarte, Efrain; Urrea, Hernan; Criollo, William; Gutierrez, Oscar

    2010-02-01

    Meningioma cell cultures were prepared from frozen cell samples in 96 wells culture plates. Semiconductor light sources (LED) in seven different wavelength ranges were used to illuminate the wells, three different irradiation doses were selected per LED. Control cultures using three different concentrations of FBS were processed for comparison. Cell proliferation, viability, and cytotoxicity were measured every 24 hours for 6 days, using the XTT colorimetric assay (RocheR). None of the irradiated cultures exhibit cytotoxicity; but some of them exhibit proliferation inhibition. The larger proliferation was detected at a 0.05J/cm2 dose, for all LEDs; but for the orange and violet LEDs generated the bigger proliferation rate was measured. Results show the improvement of meningioma cell proliferation using illumination in some given wavelength ranges.

  12. Development of a selective culture medium for bifidobacteria, Raffinose-Propionate Lithium Mupirocin (RP-MUP) and assessment of its usage with Petrifilm™ Aerobic Count plates.

    PubMed

    Miranda, Rodrigo Otávio; de Carvalho, Antonio Fernandes; Nero, Luís Augusto

    2014-05-01

    This study aimed to develop a selective culture media to enumerate bifidobacteria in fermented milk and to assess this medium when used with Petrifilm™ AC plates. For this purpose, Bifidobacterium spp., Lactobacillus spp. and Streptococcus thermophilus strains were tested to verify their fermentation patterns for different carbohydrates. All bifidobacteria strains were able to use raffinose. Based on these characteristic, a selective culture medium was proposed (Raffinose-Propionate Lithium Mupirocin, RP-MUP), used with Petrifilm™ AC plates, and was used to enumerate bifidobacteria in fermented milk. RP-MUP performance was assessed by comparing the results with this medium to reference protocols and culture media for bifidobacteria enumeration. RP-MUP, whether used or not with Petrifilm™ AC, presented similar performance to TOS-MUP (ISO 29981), with no significant differences between the mean bifidobacteria counts (p < 0.05) and with high correlation indices (r = 0.99, p < 0.05). As an advantage, reliable results were obtained after just 48 h of incubation when RP-MUP was used with Petrifilm™ AC, instead of the 72 h described in the ISO 29981 protocol. PMID:24387858

  13. A microELISA assay for detection of anti-HLA activity of mouse monoclonal antibodies using an Astroscan 2100 automated plate reader.

    PubMed

    Sadler, A M; Krausa, P; Marsh, S G; Heyes, J M; Bodmer, J G

    1992-04-27

    A microenzyme-linked immunosorbent assay (microELISA) method has been developed using an Astroscan 2100 system automated plate reader which was initially designed for tissue typing by a two colour fluorescent microcytotoxicity assay. A 96-well plate ELISA used for screening mouse monoclonal antibodies raised against surface HLA antigens has been modified for use with the Astroscan plate reader and 72-well typing trays. The existing substrate 4-methylumbelliferyl-beta-D-galactoside (4MUG) has been replaced with fluorescein-di-beta-D-galactopyranoside (FDG), to provide a wavelength (530 nm) detectable by the Astroscan or other automated plate readers designed for reading microcytotoxicity assay plates. The assay volumes have also been reduced tenfold for use with Terasaki microtest plates. The assay now has the major advantage of requiring only 5 microliters of test supernatant allowing hybridomas to be screened earlier during a fusion and on a wider cell panel. The use of the large panel which includes B lymphoblastoid cell lines (B-LCL) and mouse L cell transfectants expressing HLA genes, reduces the length of time the hybridomas need to be kept in tissue culture before selection. Other advantages include the reduction in the number of target cells required, smaller volumes of reagents throughout the assay and the ability to screen cytotoxic as well as non-cytotoxic monoclonal antibodies. The sensitivity of this microELISA proved to be comparable with the original assay and so provides an efficient screening method for monoclonal antibodies. PMID:1583310

  14. The expression of transforming growth factor-beta by cultured chick growth plate chondrocytes: differential regulation by 1,25-dihydroxyvitamin D3.

    PubMed

    Farquharson, C; Law, A S; Seawright, E; Burt, D W; Whitehead, C C

    1996-05-01

    1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and transforming growth factor-beta (TGF-beta) are both important regulators of chondrocyte growth and differentiation. We report here that 1,25(OH)2D3 differentially regulates the expression of the genes for TGF-beta 1 to -beta 3 and the secretion of the corresponding proteins in cultured chick chondrocytes. Confluent growth plate chondrocytes were serum-deprived and cultured in varying concentrations of 1,25(OH)2D3. Cells were assayed for TGF-beta mRNA and conditioned medium was assayed for TGF-beta activity and isoform composition. Active TGF-beta was only detected in 10(-8) M 1,25(OH)2D3-treated cultures (8.37 ng active TGF-beta/mg protein). There was a significant decrease in total (latent-active) TGF-beta activity in conditioned medium of 10(-12) M (23.4%; P < 0.05) and 10(-10) M (20.7%; P < 0.05) 1,25(OH)2D3-treated cultures but 10(-8) M 1,25(OH)2D3 significantly increased (30.9%; P < 0.01) TGF-beta activity. The amounts of TGF-beta 1, -beta 2 and -beta 3 isoforms produced were similar in control, 10(-10) or 10(-12) M 1,25(OH)2D3-treated cultures but the conditioned medium of 10(-8) M 1,25(OH)2D3-treated cultures contained significantly higher amounts of all three isoforms. Quantification of TGF-beta mRNA demonstrated differential control of TGF-beta gene expression with TGF-beta 1 and -beta 3 mRNA levels reduced by all concentrations of 1,25(OH)2D3 examined (10(-8), 10(-10) and 10(-12) M) whilst TGF-beta 2 mRNA concentrations were elevated. Our results indicated that 1,25(OH)2D3 regulates chick growth plate chondrocyte TGF-beta secretion and mRNA expression in a concentration-dependent and isoform-specific manner. This interaction may be important in the regulation of chondrocyte metabolism and endochondral bone growth. PMID:8708539

  15. Rickettsia rickettsii isolation from naturally infected Amblyomma parvum ticks by centrifugation in a 24-well culture plate technique

    PubMed Central

    Dzul-Rosado, K.; Peniche-Lara, G.; Tello-Martín, R.; Zavala-Velázquez, J.; Pacheco, R. de Campos; Labruna, M.B.; Sánchez, E.C.; Zavala-Castro, J.

    2013-01-01

    Rocky Mountain spotted fever is an acute illness caused by Rickettsia rickettsii (R. rickettsii) and is transmitted by the bite of ticks of the genera Dermacentor, Amblyomma and Rhipicephalus. The illness results in a high mortality rate and may be easily confused with other febrile syndromes. In Yucatan State, Mexico, childhood cases with a high mortality have been reported. In this work we report the isolation of a Mexican R. rickettsii strain from a tick egg mass using an alternative method for Rickettsia isolation with 24-well plates. We also identified a potential vector of R. rickettsii in the southeast of Mexico, which is Amblyomma parvum. PMID:26623321

  16. Twenty-four well plate miniature bioreactor system as a scale-down model for cell culture process development.

    PubMed

    Chen, Aaron; Chitta, Rajesh; Chang, David; Amanullah, Ashraf

    2009-01-01

    Increasing the throughput and efficiency of cell culture process development has become increasingly important to rapidly screen and optimize cell culture media and process parameters. This study describes the application of a miniaturized bioreactor system as a scaled-down model for cell culture process development using a CHO cell line expressing a recombinant protein. The microbioreactor system (M24) provides non-invasive online monitoring and control capability for process parameters such as pH, dissolved oxygen (DO), and temperature at the individual well level. A systematic evaluation of the M24 for cell culture process applications was successfully completed. Several challenges were initially identified. These included uneven gas distribution in the wells due to system design and lot to lot variability, foaming issues caused by sparging required for active DO control, and pH control limitation under conditions of minimal dissolved CO2. A high degree of variability was found which was addressed by changes in the system design. The foaming issue was resolved by addition of anti-foam, reduction of sparge rate, and elimination of DO control. The pH control limitation was overcome by a single manual liquid base addition. Intra-well reproducibility, as indicated by measurements of process parameters, cell growth, metabolite profiles, protein titer, protein quality, and scale-equivalency between the M24 and 2 L bioreactor cultures were very good. This evaluation has shown feasibility of utilizing the M24 as a scale-down tool for cell culture application development under industrially relevant process conditions. PMID:18683260

  17. Comparison of PCR-DGGE and selective plating methods for monitoring the dynamics of a mixed culture population in synthetic brewery wastewater.

    PubMed

    Tam, Kawai; Yang, Ching-Hong; Matsumoto, Mark R; Crowley, David E; Sheppard, John D

    2005-01-01

    Enrichment of an activated sludge inoculum in synthetic brewery wastewater, which included glucose, maltose, and ethanol, was conducted in batch experiments to identify the dominant microbes present, to determine methodologies capable of monitoring the mixed culture population dynamics, and to determine the consortium's substrate degradation behavior. These results and methodologies were subsequently used in the determination of the population dynamics of suspended and attached microorganisms in a sequencing batch system in the second part of this research work. The three-membered microbial community comprised two bacterial and one fungal species that were identified as Acinetobacter sp., Enterobacter sp., and Candida sp. PCR-DGGE and plating on selective media were used to track the population dynamics of the consortium during the degradation of different substrates in synthetic wastewater containing glucose, maltose, and ethanol. Enterobacter sp. could degrade glucose and maltose but not ethanol, whereas Acinetobacter and Candida could degrade all three carbon sources. In buffered batch mixed culture experiments, Enterobacter was the predominant bacterium until the sugar concentrations decreased to levels that enabled Acinetobacter and Candida to degrade ethanol. PCR-DGGE was effective for detecting the dominant species, but culture-based methods were more accurate for monitoring the population dynamics of these microorganisms during growth in the wastewater medium. PMID:15932247

  18. Effect of sprinkler pressure and spray plate on culturable microorganism concentrations during simulated irrigation of dairy wastewater

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study we conducted simulated spray irrigation events of dairy wastewater to assess the impact of pressure and sprinkler type upon post-sprinkler culturable microorganism concentrations. Dairy wastewater was sampled before and after it was pumped through sprinklers typically used on center p...

  19. The effect of automobile exhaust particulates on cell viability, plating efficiency and cell division of mammalian tissue culture cells.

    PubMed

    Seemayer, N H; Hadnagy, W; Tomingas, R

    1987-03-01

    Extract of particulate matter (EPM) of gasoline engine exhaust induced only a slight loss of cell viability of mouse macrophages (line IC-21) in vitro, while a strong dose-dependent reduction of plating efficiency of human cell line A-549 and of Syrian hamster line 14-1b occurred. Cytological investigations of exposed macrophages of line IC-21 revealed an increase in the mitotic index from 1.5% of control values up to 14.6% at the highest tested concentration of EPM. Mitotic arrest is based almost exclusively on C-type mitoses occurring dose-dependently in the presence of EPM. Results indicate disturbances of the spindle apparatus in the presence of EPM. PMID:2437649

  20. Validation of a sensitive and automated 96-well solid-phase extraction liquid chromatography-tandem mass spectrometry method for the determination of desloratadine and 3-hydroxydesloratadine in human plasma.

    PubMed

    Yang, Liyu; Clement, Robert P; Kantesaria, Bhavna; Reyderman, Larisa; Beaudry, Francis; Grandmaison, Charles; Di Donato, Lorella; Masse, Robert; Rudewicz, Patrick J

    2003-07-25

    To support clinical development, a liquid chromatographic-tandem mass spectrometric (LC-MS-MS) method was developed and validated for the determination of desloratadine (descarboethoxyloratadine) and 3-OH desloratadine (3-hydroxydescarboethoxyloratadine) concentrations in human plasma. The method consisted of automated 96-well solid-phase extraction for sample preparation and liquid chromatography/turbo ionspray tandem mass spectrometry for analysis. [2H(4)]Desloratadine and [2H(4)]3-OH desloratadine were used as internal standards (I.S.). A quadratic regression (weighted 1/concentration(2)) gave the best fit for calibration curves over the concentration range of 25-10000 pg/ml for both desloratadine and 3-OH desloratadine. There was no interference from endogenous components in the blank plasma tested. The accuracy (%bias) at the lower limit of quantitation (LLOQ) was -12.8 and +3.4% for desloratadine and 3-OH desloratadine, respectively. The precision (%CV) for samples at the LLOQ was 15.1 and 10.9% for desloratadine and 3-OH desloratadine, respectively. For quality control samples at 75, 1000 and 7500 pg/ml, the between run %CV was

  1. Plate motion

    SciTech Connect

    Gordon, R.G. )

    1991-01-01

    The motion of tectonic plates on the earth is characterized in a critical review of U.S. research from the period 1987-1990. Topics addressed include the NUVEL-1 global model of current plate motions, diffuse plate boundaries and the oceanic lithosphere, the relation between plate motions and distributed deformations, accelerations and the steadiness of plate motions, the distribution of current Pacific-North America motion across western North America and its margin, plate reconstructions and their uncertainties, hotspots, and plate dynamics. A comprehensive bibliography is provided. 126 refs.

  2. Data on keratin expression in human cells cultured with Australian native plant extracts.

    PubMed

    Adams, Damian H; Shou, Qingyao; Wohlmuth, Hans; Cowin, Allison J

    2016-06-01

    Australian native plants have a long history of therapeutic use in indigenous cultures particularly for the treatment of wounds. We analysed 14 plant derived compounds from the species Pilidiostigma glabrum, Myoporum montanum, Geijera parviflora, and Rhodomyrtus psidioides for keratin 1, 5, 10 and 14 supporting the research article "Native Australian plant extracts differentially induce Collagen I and Collagen III in vitro and could be important targets for the development of new wound healing therapies" [5]. An in situ immunofluorescence assay was used in a 96 well tissue culture plate format to measure keratin expression in immortalised human keratinocytes (HaCaTs) exposed Australian native plant compounds to NMR spectra for the plant extracts are included in this article as is quantitative fluorescent intensity data of keratin 1, 5, 10 and 14 expression. PMID:27077086

  3. Rapid Identification of Bacteria Directly from Positive Blood Cultures by Use of a Serum Separator Tube, Smudge Plate Preparation, and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

    PubMed

    Chen, Yan; Porter, Vanessa; Mubareka, Samira; Kotowich, Leona; Simor, Andrew E

    2015-10-01

    We analyzed the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) of smudge plate growth for bacterial identification from 400 blood cultures. Ninety-seven percent of Gram-negative bacilli and 85% of Gram-positive organisms were correctly identified within 4 h; only eight isolates (2.0%) were misidentified. This method provided rapid and accurate microbial identification from positive blood cultures. PMID:26202115

  4. Three-dimensional cell culturing by magnetic levitation for evaluating efficacy/toxicity of photodynamic therapy

    NASA Astrophysics Data System (ADS)

    Sabino, Luis G.; Menezes, Priscila F. C.; Bagnato, Vanderlei S.; Souza, Glauco; Killian, Thomas C.; Kurachi, Cristina

    2014-03-01

    We used three dimensional cell cultures (3D) based on the magnetic levitation method (MLM) to evaluate cytotoxicity of photodynamic therapy (PDT). First, we decorated Hep G2 and MDA-MB-321 cells with NanoShuttle by introducing it in the media and incubated overnight. Next day, we transferred the cells to a 6-well plate and placed a magnetic driver on the top of the plate to start levitation. We monitored the formation of the 3D cell culture by optical microscopy and after four days, we added the photosensitizer Photogem (PG) in the culture media in concentrations of 50, 25, 12.5, 6.25μg/ml. We incubated them for 24 hours, after that we washed the cultures with PBS and added fresh media. Samples were then illuminated for 600s using a 630nm LED-based device, generating light intensities of 30 mW/cm2 in a total light fluence of 18 J/cm2. Following the illumination, we added fresh media, and 30 hours later, the 3D structures were broken using a pipettor and the cells seeded in 96 well plates, 105 cells per well, with a magnetic drive placed on the bottom of the plate to create cell culture dots. After 24 hours, we used a MTT assay to evaluate PDT cytotoxicity. The PDT effect, evaluated by the half maximal effective concentration (EC50), in MDA-MB-231 cells (EC50 =3.14 μg/ml) is more aggressive compared to the effect of PDT in Hep G2 cells (EC50 = 7.48 μg/ml). It suggests that the cell culture structure and its interaction facilitated the PG uptake and consequently elevated the Photodynamic effect for MDA-MB-231.

  5. Culture.

    ERIC Educational Resources Information Center

    1997

    Twelve conference papers on cultural aspects of second language instruction include: "Towards True Multiculturalism: Ideas for Teachers" (Brian McVeigh); Comparing Cultures Through Critical Thinking: Development and Interpretations of Meaningful Observations" (Laurel D. Kamada); "Authority and Individualism in Japan and the USA" (Alisa Woodring);…

  6. Characterization of microfluidic human epidermal keratinocyte culture

    PubMed Central

    O’Neill, Adrian T.; Monteiro-Riviere, Nancy A.

    2008-01-01

    Human epidermal keratinocytes (HEK) are skin cells of primary importance in maintaining the body’s defensive barrier and are used in vitro to assess the irritation potential and toxicity of chemical compounds. Microfluidic systems hold promise for high throughput irritant and toxicity assays, but HEK growth kinetics have yet to be characterized within microscale culture chambers. This research demonstrates HEK patterning on microscale patches of Type I collagen within microfluidic channels and maintenance of these cells under constant medium perfusion for 72 h. HEK were shown to maintain 93.0%–99.6% viability at 72 h under medium perfusion ranging from 0.025–0.4 μl min−1. HEK maintained this viability while ∼100% confluent—a level not possible in 96 well plates. Microscale HEK cultures offer the ability to precisely examine the morphology, behavior and viability of individual cells which may open the door to new discoveries in toxicological screening methods and wound healing techniques. PMID:19002858

  7. Miniaturized plate readers for low-cost, high-throughput phenotypic screening.

    PubMed

    Jensen, Paul A; Dougherty, Bonnie V; Moutinho, Thomas J; Papin, Jason A

    2015-02-01

    We present a miniaturized plate reader for measuring optical density in 96-well plates. Our standalone reader fits in most incubators, environmental chambers, or biological containment suites, allowing users to leverage their existing laboratory infrastructure. The device contains no moving parts, allowing an entire 96-well plate to be read several times per second. We demonstrate how the fast sampling rate allows our reader to detect small changes in optical density, even when the device is placed in a shaking incubator. A wireless communication module allows remote monitoring of multiple devices in real time. These features allow easy assembly of multiple readers to create a scalable, accurate solution for high-throughput phenotypic screening. PMID:25366331

  8. Miniaturized Plate Readers for Low-Cost, High-Throughput Phenotypic Screening

    PubMed Central

    Jensen, Paul A.; Dougherty, Bonnie V.; Moutinho, Thomas J.

    2015-01-01

    We present a miniaturized plate reader for measuring optical density in 96-well plates. Our standalone reader fits in most incubators, environmental chambers, or biological containment suites, allowing users to leverage their existing laboratory infrastructure. The device contains no moving parts, allowing an entire 96-well plate to be read several times per second. We demonstrate how the fast sampling rate allows our reader to detect small changes in optical density, even when the device is placed in a shaking incubator. A wireless communication module allows remote monitoring of multiple devices in real time. These features allow easy assembly of multiple readers to create a scalable, accurate solution for high-throughput phenotypic screening. PMID:25366331

  9. Use of a Fluorometric Imaging Plate Reader in high-throughput screening

    NASA Astrophysics Data System (ADS)

    Groebe, Duncan R.; Gopalakrishnan, Sujatha; Hahn, Holly; Warrior, Usha; Traphagen, Linda; Burns, David J.

    1999-04-01

    High-throughput screening (HTS) efforts at Abbott Laboratories have been greatly facilitated by the use of a Fluorometric Imaging Plate Reader. The FLIPR consists of an incubated cabinet with integrated 96-channel pipettor and fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorometer. An argon laser is used to excite fluorophores in a 96-well microtiter plate and the emitted fluorescence is imaged by a cooled CCD camera. The image data is downloaded from the camera and processed to average the signal form each well of the microtiter pate for each time point. The data is presented in real time on the computer screen, facilitating interpretation and trouble-shooting. In addition to fluorescence, the camera can also detect luminescence form firefly luciferase.

  10. Real and Virtual Heritage Historical Astronomical Plate Archives in Sonneberg, Bamberg and Hamburg Observatories, the Evolution of Astrophysics and their Influence on Human Knowledge and Culture

    NASA Astrophysics Data System (ADS)

    Kunzmann, Björn

    The rise of astrophysics around 1860 introduced new instruments, methods and research areas. Of course, the increasing number of foundations of new observatories around the world starting at that time was forced by that new scientific discipline, too, but especially by the usage of photographic instruments. At the end of the 19th century the formation and development of photographic methods and techniques had reached a level of sufficient stability for productive usage in astronomy and astrophysics, their new instrumrents, methods and goals. The fundamental meaning of star light analysis for astrophysics by increasing discoveries of Variable Stars and the Systematic search for moving Solar System objects had basically driven the beginning of large photographic sky patrols at that time, using photographic glass plates as detectors and information storages. Sky Patrols, especially systematic long-term monitoring of the whole sky or of well defined selected areas and Sky Surveys were (and still are) an important key method that forced the evolution and progress of astrophysics. Important scientific results by famous astronomers, for example Walter Baade, Cuno Hoffmeister and Harlow Shapley depended on the analysis of photographic plates. Today, there are around 50 photographic plate archives world-wide. Most of them, unfortunately, are in a quite poor condition and not yet digitized. Following Harvard College Observatory with an estimated total of 600,000 plates, Sonneberg observatory harbours the second largest archive world-wide (around 300,000 plates) among other large ones in Germany like Bamberg (40,000 plates) and Hamburg (35,000 plates). These plate archives form an important heritage with a total of roughly two million direct plates and some ten or hundred thousands of spectroscopic plates. A lot of progress has been made by transforming this real heritage to a virtual one by systematic digitisation of the plates, but perhaps only 15% of them have been

  11. Peen plating

    NASA Technical Reports Server (NTRS)

    Babecki, A. J. (Inventor); Haehner, C. L.

    1973-01-01

    A process for metal plating which comprises spraying a mixture of metallic powder and small peening particles at high velocity against a surface is described. The velocity must be sufficient to impact and bond metallic powder onto the surface. In the case of metal surfaces, the process has as one of its advantages providing mechanical working (hardening) of the surface simultaneously with the metal plating.

  12. Evaluation of the cytotoxicity effect of dimethyl sulfoxide (DMSO) on Caco2/TC7 colon tumor cell cultures.

    PubMed

    Da Violante, Georges; Zerrouk, Naima; Richard, Isabelle; Provot, Gérard; Chaumeil, Jean Claude; Arnaud, Philippe

    2002-12-01

    Dimethyl sulfoxide (DMSO) is usually used to solubilize poorly soluble drugs in permeation assays such as that using Caco2 enterocyte-like cells. The objective of this study was to evaluate the toxicity of DMSO on Caco2/TC7 cells and determinate the maximal concentration usable in permeation experiments. Caco2/TC7 cells were cultured for 21 d on 96-well plates for evaluation of toxicity. The determination of lactate dehydrogenase (LDH) release in cell supernatant and the measurement of Neutral Red (NR) uptake are used for cytotoxicity assays. DMSO solutions (0-100%) in Hank's balanced salt solution containing HEPES (25 mM), pH 7.4, were incubated with Caco-2/TC7 cells on 96 well plates. Caco2/TC7 cells were cultured on Transwell-Clear inserts to evaluate the influence of DMSO on the apparent permeability of the paracellular marker mannitol. DMSO 10% did not induce any significant increase in LDH release whereas a significant increase in LDH activity (ANOVA, p<0.05) occurred at a DMSO concentration of 20 to 50%. NR incorporation in viable cells was statistically reduced by 27 to 36% at DMSO concentration of 20% up to 100% (ANOVA, p>0.05). No statistical difference (p<0.05) in apparent mannitol permeability was observed between the control and 10% DMSO groups. In conclusion, at concentrations of up to 10%, DMSO did not produce any significant alteration in apical membrane permeability or on cell-to-cell tight junctional complexes. PMID:12499647

  13. Enumeration of total yeasts and molds in foods by the SimPlate Yeast and Mold-Color Indicator method and conventional culture methods: collaborative study.

    PubMed

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

    2003-01-01

    The relative effectiveness of the SimPlate Yeast and Mold-Color Indicator method (Y&M-CI) was compared to the U.S. Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM) method and the proposed International Organization for Standardization (ISO) method, ISO/CD 21527, for enumerating yeasts and molds in foods. Test portions were prepared and incubated according to the conditions stated in both the BAM and ISO methods. Six food types were analyzed: frozen corn dogs, nut meats, frozen fruits, cake mix, cereal, and fresh cheese. Nut meats, frozen fruits, and fresh cheese were naturally contaminated. All other foods were artificially contaminated with either a yeast or mold. Seventeen 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 between the SimPlate method and the 2 corresponding reference methods. Moreover, mean log counts between the 2 reference methods were also very similar. The repeatability (Sr) and reproducibility (SR) standard deviations were comparable between the 3 method comparisons. These results indicate that the BAM method and the SimPlate method are equivalent for enumerating yeast and mold populations in foods. Similarly, the SimPlate method is comparable to the proposed ISO method when test portions are prepared and incubated as defined in the proposed ISO method. PMID:12723916

  14. Hyperspectral imaging for differentiating colonies of non-O157 shiga-toxin producing echerichia coli (STEC) serogroups on spread plates of pure cultures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Direct plating onto solid agar media has been widely used in microbiology laboratories for presumptive-positive pathogen detection in spite of the fact that it is often subjective, labor intensive and time consuming. Rainbow agar is a selective and chromogenic medium that helps to detect pathogenic ...

  15. A Compact Immunoassay Platform Based on a Multicapillary Glass Plate

    PubMed Central

    Xue, Shuhua; Zeng, Hulie; Yang, Jianmin; Nakajima, Hizuru; Uchiyama, Katsumi

    2014-01-01

    A highly sensitive, rapid immunoassay performed in the multi-channels of a micro-well array consisting of a multicapillary glass plate (MCP) and a polydimethylsiloxane (PDMS) slide is described. The micro-dimensions and large surface area of the MCP permitted the diffusion distance to be decreased and the reaction efficiency to be increased. To confirm the concept of the method, human immunoglobulin A (h-IgA) was measured using both the proposed immunoassay system and the traditional 96-well plate method. The proposed method resulted in a 1/5-fold decrease of immunoassay time, and a 1/56-fold cut in reagent consumption with a 0.05 ng/mL of limit of detection (LOD) for IgA. The method was also applied to saliva samples obtained from healthy volunteers. The results correlated well to those obtained by the 96-well plate method. The method has the potential for use in disease diagnostic or on-site immunoassays. PMID:24859022

  16. 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

  17. Assessment of a new selective chromogenic Bacillus cereus group plating medium and use of enterobacterial autoinducer of growth for cultural identification of Bacillus species.

    PubMed

    Reissbrodt, R; Rassbach, A; Burghardt, B; Rienäcker, I; Mietke, H; Schleif, J; Tschäpe, H; Lyte, M; Williams, P H

    2004-08-01

    A new chromogenic Bacillus cereus group plating medium permits differentiation of pathogenic Bacillus species by colony morphology and color. Probiotic B. cereus mutants were distinguished from wild-type strains by their susceptibilities to penicillin G or cefazolin. The enterobacterial autoinducer increased the sensitivity and the speed of enrichment of B. cereus and B. anthracis spores in serum-supplemented minimal salts medium (based on the standard American Petroleum Institute medium) and buffered peptone water. PMID:15297532

  18. One plate, two plates, a thousand plates. How crystallisation changes with large numbers of samples.

    PubMed

    Newman, Janet

    2011-09-01

    Turning commercial lab automation into a high-throughput centre requires an underlying process, and implementing checks to ensure that the process is working as it should. At the Collaborative Crystallisation Centre (C3), protein samples from local, national and international groups are set up in crystallisation screening and optimisation experiments with two thousand 96 well plates being set up each year. During its five years of operation, the C3 has implemented a series of enabling protocols - from simple 'reality checks' to determine if a screen has evaporated during storage to more sophisticated systems such as a sample labelling and tracking system. The most important - and perhaps surprising - lesson has been how much effort is required to effectively communicate between the centre and its clients, as well as between the centre's staff members. It is easy to confuse the concept of 'high throughput' in any field with the idea of setting up an experiment quickly. Although automation can be used to set up a single experiment more rapidly than can be done by hand, the distinguishing feature of a high throughput technology is the sustainability of the increased rate. PMID:21571072

  19. Multiplexed chemiluminescent assays in ArrayPlates for high-throughput measurement of gene expression

    NASA Astrophysics Data System (ADS)

    Martel, Ralph R.; Rounseville, Matthew P.; Botros, Ihab W.; Seligmann, Bruce E.

    2002-06-01

    Multiplexed Molecular Profiling (MMP) assays for drug discovery are performed in ArrayPlates. ArrayPlates are 96- well microtiter plates that contain a 16-element array at the bottom of each well. Each element within an array measures one analyte in a sample. A CCD imager records the quantitative chemiluminescent readout of all 1,536 elements in a 96-well plate simultaneously. Since array elements are reagent modifiable by the end-user, ArrayPlates can be adapted to a broad range of nucleic acid- and protein-based assays. Such multiplexed assays are rapidly established, flexible, robust, automation-friendly and cost-effective. Nucleic acid assays in ArrayPlates can detect DNA and RNA, including SNPs and ESTs. A multiplexed mRNA assay to measure the expression of 16 genes is described. The assay combines a homogeneous nuclease protection assay with subsequent probe immobilization to the array by means of a sandwich hybridization followed with chemiluminescent detection. This assay was used to examine cells grown and treated in microplates and avoided cloning, transfection, RNA insolation, reverse transcription, amplification and fluorochrome labeling. Standard deviations for the measurement of 16 genes ranged from 3 percent to 13 percent in samples of 30,000 cells. Such ArrayPlates transcription assays are useful in drug discovery and development for target validation, screening, lead optimization, metabolism and toxicity profiling. Chemiluminescent detection provides ArrayPlates assays with high signal-to-noise readout and simplifies imager requirements. Imaging a 2D surface that contains arrays simplifies lens requirements relative to imaging columns of liquid in microtiter plate wells. The Omix imager for ArrayPlates is described.

  20. Differentiation of big-six non-O157 shiga-toxin producing escherichia coli (STEC) on spread plates of mixed cultures using hyperspectral imaging

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There have been considerable recent advances in the technology for rapidly detecting foodborne pathogens. However, a traditional culture method is still the “gold standard” for presumptive-positive pathogen screening although it is labor-intensive, ineffective in testing large amount of food samples...

  1. Application of 16S rDNA-DGGE and plate culture to characterization of bacterial communities associated with the sawfly, Acantholyda erythrocephala (Hymenoptera, Pamphiliidae).

    PubMed

    Zahner, Viviane; Lucarotti, Christopher J; McIntosh, Douglas

    2008-12-01

    Culture-based analysis was employed in parallel with PCR amplification of 16S rDNA, coupled with denaturing gradient gel electrophoresis (DGGE), to profile bacterial species associated with different developmental stages of the pine false webworm (PFW), Acantholyda erythrocephala, a sawfly pest responsible for incidents of severe defoliation in commercially important tree plantations in North America. Culture-based analysis revealed that Pseudomonas spp. along with Bacillus sphaericus and Arthrobacter sp. were the predominant components of the microflora of the internal organs and identified life-stage-specific associations including Photorhabdus temperata with egg and larval samples and a Janthinobacterium sp. with eonymphs. PCR-DGGE confirmed the predominance of Pseudomonas spp. and B. sphaericus in the majority of samples but did not detect Arthrobacter sp., P. temperate, or Janthinobacterium sp. In contrast, DGGE revealed the presence of a Chryseobacterium sp. as the predominant component of the PFW micoflora at all life stages, with the exception of adults. This species had been infrequently cultured, at low levels, from a limited number of samples and the existence of a possible relationship between this bacterium and the PFW had gone unnoticed using the culture-based approach. Our findings highlight the advantages of applying a dual approach to the study of microbe-insect associations and demonstrate that the benefits of one system can be used to overcome some of the limitations of the other. PMID:18769850

  2. Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture

    PubMed Central

    Luni, Camilla; Feldman, Hope C.; Pozzobon, Michela; De Coppi, Paolo; Meinhart, Carl D.; Elvassore, Nicola

    2010-01-01

    This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100–300 μl). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for stem cell production at the clinical level. The controlled stirring inside the wells of a standard 96-well plate is provided by buoyancy-driven thermoconvection. The temperature and velocity fields within the culture volume are determined with numerical simulations. The numerical results are verified with experimental velocity measurements using microparticle image velocimetry (μPIV) and are used to define feasible experimental conditions for stem cell cultures. To test the bioreactor array’s functionality, human umbilical cord blood-derived CD34+ cells were cultured for 7 days at five different stirring conditions (0.24–0.58 μm∕s) in six repeated experiments. Cells were characterized in terms of proliferation, and flow cytometry measurements of viability and CD34 expression. The microliter-bioreactor array demonstrates its ability to support HSC cultures under stirred conditions without adversely affecting the cell behavior. Because of the highly controlled operative conditions, it can be used to explore culture conditions where the mass transport of endogenous and exogenous growth factors is selectively enhanced, and cell suspension provided. While the bioreactor array was developed for culturing HSCs, its application can be extended to other cell types. PMID:20824067

  3. Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies.

    PubMed

    Cheng, Ta-Chun; Pan, Chu-Hsiang; Chen, Chien-Shu; Chuang, Kuo-Hsiang; Chuang, Chih-Hung; Huang, Chien-Chaio; Chu, Yu-Yi; Yang, Ya-Chun; Chu, Pei-Yu; Kao, Chien-Han; Hsieh, Yuan-Chin; Cheng, Tian-Lu

    2015-07-01

    The envelope glycoprotein E2 of classical swine fever virus (CSFV) is widely used as a marker for measuring vaccine efficacy and antibody titer. The glycosylation profile of E2 may affect the immunogenicity of the vaccine and the timing of re-vaccination. In this study, a human embryonic kidney cell line was used to secrete fully-glycosylated CSFV E2, which was then coated onto ELISA plates without purification or adjustment. The resulting E2-secreting medium-direct-coating (E2-mDc) ELISA was successfully used to measure anti-E2 antibody titers in vaccinated and field pig sera samples. Compared with a virus neutralization test (as standard), the E2-mDc ELISA was found to be more accurate (90%) than a commercial CSFV antibody diagnostic kit (62%). In conclusion, the mammalian cell-secreted antigen can provide cheap, accurate and effective assays for vaccine efficacy and disease diagnoses. PMID:25975854

  4. Monolayer culturing and cloning of human pluripotent stem cells on laminin-521-based matrices under xeno-free and chemically defined conditions.

    PubMed

    Rodin, Sergey; Antonsson, Liselotte; Hovatta, Outi; Tryggvason, Karl

    2014-10-01

    A robust method for culturing human pluripotent stem (hPS) cells under chemically defined and xeno-free conditions is an important tool for stem cell research and for the development of regenerative medicine. Here, we describe a protocol for monolayer culturing of Oct-4-positive hPS cells on a specific laminin-521 (LN-521) isoform, under xeno-free and chemically defined conditions. The cells are dispersed into single-cell suspension and then plated on LN-521 isoform at densities higher than 5,000 cells per cm², where they attach, migrate and survive by forming small monolayer cell groups. The cells avidly divide and expand horizontally until the entire dish is covered by a confluent monolayer. LN-521, in combination with E-cadherin, allows cloning of individual hPS cells in separate wells of 96-well plates without the presence of rho-associated protein kinase (ROCK) inhibitors or any other inhibitors of anoikis. Characterization of cells maintained for several months in culture reveals pluripotency with a minimal degree of genetic abnormalities. PMID:25211513

  5. Cytokine and Eicosanoid Production by Cultured Human Monocytes Exposed to Titanium Particulate Debris

    NASA Astrophysics Data System (ADS)

    Robinson, Timothy M.; Manley, Paul A.; Sims, Paul A.; Albrecht, Ralph; Darien, Benjamin J.

    1999-10-01

    Phagocytosis of particulate wear debris from arthroplasties by macrophages induces an inflammatory response that has been linked to implant loosening and premature failure of artificial joints. Inflammatory mediators released by phagocytic macrophages such as tumor necrosis factor-a (TNF-[alpha]), interleukin-1[beta] (IL-1[beta]), interleukin-6 (IL-6), and prostaglandin E2 (PGE2) are believed to play a central role in the pathogenesis of aseptic loosening. The objective of this study was to characterize titanium alloy particulates that closely match wear debris found around joint arthroplasties and to study their effects on the biosynthesis of inflammatory mediators by cultured monocytes. Peripheral blood monocytes were isolated from healthy human volunteers. Monocytes were cultured in 96-well plates for 24 h, washed, and exposed to three concentrations of titanium particulates and controls from 18Ð24 h. Supernatants were assayed for TNF-[alpha], IL-1[beta], IL-6, and PGE2 activity. Energy dispersive X-ray spectroscopy (EDX) verified the titanium alloy to be Ti6A14V. Scanning electron microscopy (SEM) analysis showed significant titanium particulate heterogeneity with approximately 95% of the particles <1 micrometer in diameter. SEM and EDX technology was useful in the characterization of the titanium particulates utilized for in vitro models of titanium-induced cytokine release by monocytes. Incubation of titanium particulates (in concentrations similar to those found around loosened prosthetic joints) with cultured monocytes significantly increased their production of TNF-[alpha], IL-1[beta], and PGE2.

  6. Controllable Production of Transplantable Adult Human High-Passage Dermal Papilla Spheroids Using 3D Matrigel Culture

    PubMed Central

    Miao, Yong; Sun, Ya Bin; Liu, Bing Cheng; Jiang, Jin Dou

    2014-01-01

    We have succeeded in culturing human dermal papilla (DP) cell spheroids and developed a three-dimensional (3D) Matrigel (basement membrane matrix) culture technique that can enhance and restore DP cells unique characteristics in vitro. When 1×104 DP cells were cultured on the 96-well plates precoated with Matrigel for 5 days, both passage 2 and passage 8 DP cells formed spheroidal microtissues with a diameter of 150–250 μm in an aggregative and proliferative manner. We transferred and recultured these DP spheroids onto commercial plates. Cells within DP spheres could disaggregate and migrate out, which was similar to primary DP. Moreover, we examined the expression of several genes and proteins associated with hair follicle inductivity of DP cells, such as NCAM, Versican, and α-smooth muscle actin, and confirmed that their expression level was elevated in the spheres compared with the dissociated DP cells. To examine the hair-inducing ability of DP spheres, hair germinal matrix cells (HGMCs) and DP spheres were mixed and cultured on Matrigel. Unlike the dissociated DP cells and HGMCs cocultured in two dimensions, HGMCs can differentiate into hair-like fibers under the induction of the DP spheres made from the high-passage cells (passage 8) in vitro. We are the first to show that passage 3 human HGMCs differentiate into hair-like fibers in the presence of human DP spheroids. These results suggest that the 3D Matrigel culture technique is an ideal culture model for forming DP spheroids and that sphere formation partially models the intact DP, resulting in hair induction, even by high-passage DP cells. PMID:24528213

  7. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing.

    PubMed

    Riba, J; Gleichmann, T; Zimmermann, S; Zengerle, R; Koltay, P

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1 μm and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35 pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20 μm in size. Further, the magnification of the optical system used for cell detection was increased. Redesign of the optical path allows for collision-free addressing of any flat substrate since no compartment protrudes below the nozzle of the dispenser chip anymore. The improved system allows for deterministic isolation of individual bacterial cells. A single-cell printing efficiency of 93% was obtained as shown by printing fluorescent labeled E. coli. A 96-well plate filled with growth medium is inoculated with single bacteria cells on average within about 8 min. Finally, individual bacterial cells from a heterogeneous sample of E. coli and E. faecalis were isolated for clonal culturing directly on agar plates in user-defined array geometry. PMID:27596612

  8. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing

    PubMed Central

    Riba, J.; Gleichmann, T.; Zimmermann, S.; Zengerle, R.; Koltay, P.

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1 μm and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35 pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20 μm in size. Further, the magnification of the optical system used for cell detection was increased. Redesign of the optical path allows for collision-free addressing of any flat substrate since no compartment protrudes below the nozzle of the dispenser chip anymore. The improved system allows for deterministic isolation of individual bacterial cells. A single-cell printing efficiency of 93% was obtained as shown by printing fluorescent labeled E. coli. A 96-well plate filled with growth medium is inoculated with single bacteria cells on average within about 8 min. Finally, individual bacterial cells from a heterogeneous sample of E. coli and E. faecalis were isolated for clonal culturing directly on agar plates in user-defined array geometry. PMID:27596612

  9. 3D cell culture and osteogenic differentiation of human bone marrow stromal cells plated onto jet-sprayed or electrospun micro-fiber scaffolds.

    PubMed

    Brennan, Meadhbh Á; Renaud, Audrey; Gamblin, Anne-Laure; D'Arros, Cyril; Nedellec, Steven; Trichet, Valerie; Layrolle, Pierre

    2015-08-01

    A major limitation of the 2D culture systems is that they fail to recapitulate the in vivo 3D cellular microenvironment whereby cell-cell and cell-extracellular matrix (ECM) interactions occur. In this paper, a biomaterial scaffold that mimics the structure of collagen fibers was produced by jet-spraying. This micro-fiber polycaprolactone (PCL) scaffold was evaluated for 3D culture of human bone marrow mesenchymal stromal cells (MSCs) in comparison with a commercially available electrospun scaffold. The jet-sprayed scaffolds had larger pore diameters, greater porosity, smaller diameter fibers, and more heterogeneous fiber diameter size distribution compared to the electrospun scaffolds. Cells on jet-sprayed constructs exhibited spread morphology with abundant cytoskeleton staining, whereas MSCs on electrospun scaffolds appeared less extended with fewer actin filaments. MSC proliferation and cell infiltration occurred at a faster rate on jet-sprayed compared to electrospun scaffolds. Osteogenic differentiation of MSCs and ECM production as measured by ALP, collagen and calcium deposition was superior on jet-sprayed compared to electrospun scaffolds. The jet-sprayed scaffold which mimics the native ECM and permits homogeneous cell infiltration is important for 3D in vitro applications such as bone cellular interaction studies or drug testing, as well as bone tissue engineering strategies. PMID:26238732

  10. Measuring Chitinase and Protease Activity in Cultures of Fungal Entomopathogens.

    PubMed

    Cheong, Peter; Glare, Travis R; Rostás, Michael; Haines, Stephen R

    2016-01-01

    Entomopathogenic fungi produce a variety of destructive enzymes and metabolites to overcome the unique defense mechanisms of insects. In a first step, fungal chitinases and proteinases need to break down the insect's cuticle. Both enzyme classes support the infection process by weakening the chitin barrier and by producing nutritional cleavage products for the fungus. In a second step, the pathogen can now mechanically penetrate the weakened cuticle and reach the insect's hemolymph where it starts proliferating. The critical enzymes chitinase and proteinase are also excreted into the supernatants of fungal cultures and can be used as indicators of virulence. Chromogenic assays adapted for 96-well microtiter plates that measure these enzymes provide a sensitive, fast, and easy screening method for evaluating the potential biocontrol activity of fungal isolates and may be considered as an alternative to laborious and time-consuming bioassays. Furthermore, monitoring fungal enzyme production in dependence of time, nutrient sources, or other factors can facilitate in establishing optimal growth and harvesting conditions for selected isolates with the aim of achieving maximum biocontrol activity. PMID:27565500

  11. Detection of Legionella species in sewage and ocean water by polymerase chain reaction, direct fluorescent-antibody, and plate culture methods.

    PubMed

    Palmer, C J; Tsai, Y L; Paszko-Kolva, C; Mayer, C; Sangermano, L R

    1993-11-01

    Legionella spp. are ubiquitous in most environmental water sources; however, sewage treatment plants have not been examined as potential environmental reservoirs for these bacteria. This study used polymerase chain reaction, direct fluorescent-antibody staining, and culture methods to examine raw and treated sewage, ocean-receiving waters, and nearshore coastal environments for the presence of Legionella pneumophila and other Legionella spp. The study concluded that Legionella spp. are present in all phases of sewage treatment and that population numbers do not significantly decline through the treatment process. Ocean-receiving waters located 5 miles offshore, where the treated sewage is discharged, were found to contain Legionella spp., but ocean water between the discharge site and coastal bathing beaches was negative. This suggests that the Legionella spp. from the ocean discharge site were not reaching the nearshore beach waters. A flood control channel and river that entered the ocean were found to contain Legionella spp., and a nearby beach swimming area was also found to be positive, suggesting that land runoff from the flood control channel and river were the source of the Legionella spp. in the beach water samples that tested positive. PMID:8285669

  12. Detection of Legionella species in sewage and ocean water by polymerase chain reaction, direct fluorescent-antibody, and plate culture methods.

    PubMed Central

    Palmer, C J; Tsai, Y L; Paszko-Kolva, C; Mayer, C; Sangermano, L R

    1993-01-01

    Legionella spp. are ubiquitous in most environmental water sources; however, sewage treatment plants have not been examined as potential environmental reservoirs for these bacteria. This study used polymerase chain reaction, direct fluorescent-antibody staining, and culture methods to examine raw and treated sewage, ocean-receiving waters, and nearshore coastal environments for the presence of Legionella pneumophila and other Legionella spp. The study concluded that Legionella spp. are present in all phases of sewage treatment and that population numbers do not significantly decline through the treatment process. Ocean-receiving waters located 5 miles offshore, where the treated sewage is discharged, were found to contain Legionella spp., but ocean water between the discharge site and coastal bathing beaches was negative. This suggests that the Legionella spp. from the ocean discharge site were not reaching the nearshore beach waters. A flood control channel and river that entered the ocean were found to contain Legionella spp., and a nearby beach swimming area was also found to be positive, suggesting that land runoff from the flood control channel and river were the source of the Legionella spp. in the beach water samples that tested positive. Images PMID:8285669

  13. In vitro co-culture model of medulloblastoma and human neural stem cells for drug delivery assessment.

    PubMed

    Ivanov, Delyan P; Parker, Terry L; Walker, David A; Alexander, Cameron; Ashford, Marianne B; Gellert, Paul R; Garnett, Martin C

    2015-07-10

    Physiologically relevant in vitro models can serve as biological analytical platforms for testing novel treatments and drug delivery systems. We describe the first steps in the development of a 3D human brain tumour co-culture model that includes the interplay between normal and tumour tissue along with nutrient gradients, cell-cell and cell-matrix interactions. The human medulloblastoma cell line UW228-3 and human foetal brain tissue were marked with two supravital fluorescent dyes (CDCFDASE, Celltrace Violet) and cultured together in ultra-low attachment 96-well plates to form reproducible single co-culture spheroids (d = 600 μm, CV% = 10%). Spheroids were treated with model cytotoxic drug etoposide (0.3-100 μM) and the viability of normal and tumour tissue quantified separately using flow cytometry and multiphoton microscopy. Etoposide levels of 10 μM were found to maximise toxicity to tumours (6.5% viability) while stem cells maintained a surviving fraction of 40%. The flexible cell marking procedure and high-throughput compatible protocol make this platform highly transferable to other cell types, primary tissues and personalised screening programs. The model's key anticipated use is for screening and assessment of drug delivery strategies to target brain tumours, and is ready for further developments, e.g. differentiation of stem cells to a range of cell types and more extensive biological validation. PMID:25592050

  14. Effects of diethylstilbestrol on the proliferation and tyrosinase activity of cultured human melanocytes

    PubMed Central

    TANG, JIANBING; LI, QIN; CHENG, BIAO; HUANG, CHONG; CHEN, KUI

    2015-01-01

    The aim of the present study was to observe the effects of different exogenous estrogen diethylstilbestrol (DES) concentrations on the human melanocyte proliferation and tyrosinase activity. Skin specimens were obtained following blepharoplasty, and the melanocytes were primary cultured and passaged to the third generation. The melanocytes were seeded in 96-well plates, each well had 5×103 cells. The medium was changed after 24 h, and contained 10−4-10−8 M DES. After the melanocytes were incubated, the proliferation and tyrosinase activity were detected by the MTT assay and L-DOPA reaction. DES (10−8-10−6 M) enhanced the proliferation of cultured melanocytes. The intensity was positively correlated with the concentration of drug. DES, >10−5 M, inhibited the melanocytes proliferation or even produced the toxicity effect. Following the addition of 10−6 M DES to the medium, the tyrosinase activity of melanocytes was significantly increased, with P<0.05. In conclusion, a certain concentration of DES promoted the proliferation of melanocytes, enhanced the activity of tyrosinase and promoted pigment synthesis of melanocytes, with the optimal concentration of 10−6 M. PMID:26171155

  15. Sputtering and ion plating

    NASA Technical Reports Server (NTRS)

    1972-01-01

    The proceedings of a conference on sputtering and ion plating are presented. Subjects discussed are: (1) concepts and applications of ion plating, (2) sputtering for deposition of solid film lubricants, (3) commercial ion plating equipment, (4) industrial potential for ion plating and sputtering, and (5) fundamentals of RF and DC sputtering.

  16. Quantitation of megakaryocytopoiesis in liquid culture by enzymatic determination of acetylcholinesterase.

    PubMed

    Burstein, S A; Boyd, C N; Dale, G L

    1985-01-01

    A method has been developed to quantitate megakaryocytopoiesis in culture by measuring acetylcholinesterase synthesized in vitro. Murine marrow cells, treated with diisopropylfluorosphosphate (DFP) to inactivate initial acetylcholinesterase (AchE) present in megakaryocytes and contaminating blood, were set up in Iscove's medium supplemented with 15% DFP-treated horse serum +/- pokeweed mitogen-stimulated spleen cell conditioned medium (PWM-SCM) in 96-well microplates. Following the culture period, Triton X-100, dithiobisnitrobenzoic acid (DTNB), and acetylthiocholine iodide were added to each well. AchE synthesized in culture cleaved acetylthiocholine to thiocholine, which stochiometrically reduced the colorless indicator DTNB to a highly colored product. Thirty minutes following the addition of substrate, the plates were assayed for activity with a vertical recording photometer. When platelets, freshly prepared bone marrow cells, or cultured marrow were assayed by this method, a linear relationship was observed between optical density (OD) and the number of cells assayed. Moreover, a linear relationship between the number of AchE-positive megakaryocytes determined histochemically and AchE activity determined spectrophotometrically was observed. Red cells exhibited no activity. Inhibitor studies demonstrated that the activity measured was true AchE. Separation of marrow by density gradient centrifugation showed that the megakaryocyte enriched fraction contained all the AchE while the megakaryocyte depleted fraction contained none. From the data we conclude that this rapid, semiautomated method quantitates megakaryocytic AchE synthesis in culture, and that this method will be a useful assay system for the detection of factors that influence megakaryocytopoiesis. PMID:3965482

  17. MerMade: An Oligodeoxyribonucleotide Synthesizer for High Throughput Oligonucleotide Production in Dual 96-Well Plates

    PubMed Central

    Rayner, Simon; Brignac, Stafford; Bumeister, Ron; Belosludtsev, Yuri; Ward, Travis; Grant, O’dell; O’Brien, Kevin; Evans, Glen A.; Garner, Harold R.

    1998-01-01

    We have designed and constructed a machine that synthesizes two standard 96-well plates of oligonucleotides in a single run using standard phosphoramidite chemistry. The machine is capable of making a combination of standard, degenerate, or modified oligos in a single plate. The run time is typically 17 hr for two plates of 20-mers and a reaction scale of 40 nm. The reaction vessel is a standard polypropylene 96-well plate with a hole drilled in the bottom of each well. The two plates are placed in separate vacuum chucks and mounted on an xy table. Each well in turn is positioned under the appropriate reagent injection line and the reagent is injected by switching a dedicated valve. All aspects of machine operation are controlled by a Macintosh computer, which also guides the user through the startup and shutdown procedures, provides a continuous update on the status of the run, and facilitates a number of service procedures that need to be carried out periodically. Over 25,000 oligos have been synthesized for use in dye terminator sequencing reactions, polymerase chain reactions (PCRs), hybridization, and RT–PCR. Oligos up to 100 bases in length have been made with a coupling efficiency in excess of 99%. These machines, working in conjunction with our oligo prediction code are particularly well suited to application in automated high throughput genomic sequencing. PMID:9685322

  18. Efficient DNA sequencing on microtiter plates using dried reagents and Bst DNA polymerase.

    PubMed

    Earley, J J; Kuivaniemi, H; Prockop, D J; Tromp, G

    1993-01-01

    Sequenase, Taq DNA polymerase and Bst DNA polymerase were tested for sequencing of DNA on microtiter plates using dried down reagents. Several parameters were investigated to expedite the drying process while minimizing damage to the enzyme. Sequenase did not tolerate drying very well, and frequently generated sequences with weak signals and many sites of premature termination. With Taq DNA polymerase it was possible to obtain sequences of good quality. However, there was considerable variation of results between experiments and between batches of microtiter plates. Bst DNA polymerase generated sequences of excellent quality. It was stable for more than a week in dried-down state at -20 degrees C and at least overnight at room temperature. The method described here using Bst DNA polymerase is well suited for laboratory robots and workstations that typically employ 96-well microtiter plates. PMID:8173079

  19. Earth's Decelerating Tectonic Plates

    SciTech Connect

    Forte, A M; Moucha, R; Rowley, D B; Quere, S; Mitrovica, J X; Simmons, N A; Grand, S P

    2008-08-22

    Space geodetic and oceanic magnetic anomaly constraints on tectonic plate motions are employed to determine a new global map of present-day rates of change of plate velocities. This map shows that Earth's largest plate, the Pacific, is presently decelerating along with several other plates in the Pacific and Indo-Atlantic hemispheres. These plate decelerations contribute to an overall, globally averaged slowdown in tectonic plate speeds. The map of plate decelerations provides new and unique constraints on the dynamics of time-dependent convection in Earth's mantle. We employ a recently developed convection model constrained by seismic, geodynamic and mineral physics data to show that time-dependent changes in mantle buoyancy forces can explain the deceleration of the major plates in the Pacific and Indo-Atlantic hemispheres.

  20. ASSEMBLY OF PARALLEL PLATES

    DOEpatents

    Groh, E.F.; Lennox, D.H.

    1963-04-23

    This invention is concerned with a rigid assembly of parallel plates in which keyways are stamped out along the edges of the plates and a self-retaining key is inserted into aligned keyways. Spacers having similar keyways are included between adjacent plates. The entire assembly is locked into a rigid structure by fastening only the outermost plates to the ends of the keys. (AEC)

  1. Accelerated plate tectonics.

    PubMed

    Anderson, D L

    1975-03-21

    The concept of a stressed elastic lithospheric plate riding on a viscous asthenosphere is used to calculate the recurrence interval of great earthquakes at convergent plate boundaries, the separation of decoupling and lithospheric earthquakes, and the migration pattern of large earthquakes along an arc. It is proposed that plate motions accelerate after great decoupling earthquakes and that most of the observed plate motions occur during short periods of time, separated by periods of relative quiescence. PMID:17799689

  2. Spiral Plate Method for Bacterial Determination

    PubMed Central

    Gilchrist, J. E.; Campbell, J. E.; Donnelly, C. B.; Peeler, J. T.; Delaney, J. M.

    1973-01-01

    A method is described for determining the number of bacteria in a solution by the use of a machine which deposits a known volume of sample on a rotating agar plate in an ever decreasing amount in the form of an Archimedes spiral. After the sample is incubated, different colony densities are apparent on the surface of the plate. A modified counting grid is described which relates area of the plate of volume of sample. By counting an appropriate area of the plate, the number of bacteria in the sample is estimated. This method was compared to the pour plate procedure with the use of pure and mixed cultures in water and milk. The results did not demonstrate a significant difference in variance between duplicates at the α = 0.01 level when concentrations of 600 to 12 × 105 bacteria per ml were used, but the spiral plate method gave counts that were higher than counts obtained by the pour plate method. The time and materials required for this method are substantially less than those required for the conventional aerobic pour plate procedure. Images PMID:4632851

  3. Selective isolation of ammonia-oxidizing bacteria from autotrophic nitrifying granules by applying cell-sorting and sub-culturing of microcolonies

    PubMed Central

    Fujitani, Hirotsugu; Kumagai, Asami; Ushiki, Norisuke; Momiuchi, Kengo; Tsuneda, Satoshi

    2015-01-01

    Nitrification is a key process in the biogeochemical nitrogen cycle and biological wastewater treatment that consists of two stepwise reactions, ammonia oxidation by ammonia-oxidizing bacteria (AOB) or archaea followed by nitrite oxidation by nitrite-oxidizing bacteria. One of the representatives of the AOB group is Nitrosomonas mobilis species. Although a few pure strains of this species have been isolated so far, approaches to their preservation in pure culture have not been established. Here, we report isolation of novel members of the N. mobilis species from autotrophic nitrifying granules used for ammonia-rich wastewater treatment. We developed an isolation method focusing on microcolonies formation of nitrifying bacteria. Two kinds of distinctive light scattering signatures in a cell-sorting system enabled to separate microcolonies from single cells and heterogeneous aggregates within granule samples. Inoculation of a pure microcolony into 96-well microtiter plates led to successful sub-culturing and increased probability of isolation. Obtained strain Ms1 is cultivated in the liquid culture with relatively high ammonia or nitrite concentration, not extremely slow growing. Considering environmental clones that were closely related to N. mobilis and detected in various environments, the availability of this novel strain would facilitate to reveal this member’s ecophysiology in a variety of habitats. PMID:26528282

  4. Plating Tank Control Software

    1998-03-01

    The Plating Tank Control Software is a graphical user interface that controls and records plating process conditions for plating in high aspect ratio channels that require use of low current and long times. The software is written for a Pentium II PC with an 8 channel data acquisition card, and the necessary shunt resistors for measuring currents in the millampere range.

  5. Rotatable shear plate interferometer

    DOEpatents

    Duffus, Richard C.

    1988-01-01

    A rotatable shear plate interferometer comprises a transparent shear plate mounted obliquely in a tubular supporting member at 45.degree. with respect to its horizontal center axis. This tubular supporting member is supported rotatably around its center axis and a collimated laser beam is made incident on the shear plate along this center axis such that defocus in different directions can be easily measured.

  6. Microfabricated glucose biosensor for culture well operation.

    PubMed

    Pemberton, R M; Cox, T; Tuffin, R; Sage, I; Drago, G A; Biddle, N; Griffiths, J; Pittson, R; Johnson, G; Xu, J; Jackson, S K; Kenna, G; Luxton, R; Hart, J P

    2013-04-15

    A water-based carbon screen-printing ink formulation, containing the redox mediator cobalt phthalocyanine (CoPC) and the enzyme glucose oxidase (GOx), was investigated for its suitability to fabricate glucose microbiosensors in a 96-well microplate format: (1) the biosensor ink was dip-coated onto a platinum (Pt) wire electrode, leading to satisfactory amperometric performance; (2) the ink was deposited onto the surface of a series of Pt microelectrodes (10-500 μm diameter) fabricated on a silicon substrate using MEMS (microelectromechanical systems) microfabrication techniques: capillary deposition proved to be successful; a Pt microdisc electrode of ≥100 μm was required for optimum biosensor performance; (3) MEMS processing was used to fabricate suitably sized metal (Pt) tracks and pads onto a silicon 96 well format base chip, and the glucose biosensor ink was screen-printed onto these pads to create glucose microbiosensors. When formed into microwells, using a 340 μl volume of buffer, the microbiosensors produced steady-state amperometric responses which showed linearity up to 5 mM glucose (CV=6% for n=5 biosensors). When coated, using an optimised protocol, with collagen in order to aid cell adhesion, the biosensors continued to show satisfactory performance in culture medium (linear range to 2 mM, dynamic range to 7 mM, CV=5.7% for n=4 biosensors). Finally, the operation of these collagen-coated microbiosensors, in 5-well 96-well format microwells, was tested using a 5-channel multipotentiostat. A relationship between amperometric response due to glucose, and cell number in the microwells, was observed. These results indicate that microphotolithography and screen-printing techniques can be combined successfully to produce microbiosensors capable of monitoring glucose metabolism in 96 well format cell cultures. The potential application areas for these microbiosensors are discussed. PMID:23265827

  7. An improved plating process

    NASA Technical Reports Server (NTRS)

    Askew, John C.

    1994-01-01

    An alternative to the immersion process for the electrodeposition of chromium from aqueous solutions on the inside diameter (ID) of long tubes is described. The Vessel Plating Process eliminates the need for deep processing tanks, large volumes of solutions, and associated safety and environmental concerns. Vessel Plating allows the process to be monitored and controlled by computer thus increasing reliability, flexibility and quality. Elimination of the trivalent chromium accumulation normally associated with ID plating is intrinsic to the Vessel Plating Process. The construction and operation of a prototype Vessel Plating Facility with emphasis on materials of construction, engineered and operational safety and a unique system for rinse water recovery are described.

  8. Multicolor printing plate joining

    NASA Technical Reports Server (NTRS)

    Waters, W. J. (Inventor)

    1984-01-01

    An upper plate having ink flow channels and a lower plate having a multicolored pattern are joined. The joining is accomplished without clogging any ink flow paths. A pattern having different colored parts and apertures is formed in a lower plate. Ink flow channels each having respective ink input ports are formed in an upper plate. The ink flow channels are coated with solder mask and the bottom of the upper plate is then coated with solder. The upper and lower plates are pressed together at from 2 to 5 psi and heated to a temperature of from 295 F to 750 F or enough to melt the solder. After the plates have cooled and the pressure is released, the solder mask is removed from the interior passageways by means of a liquid solvent.

  9. Simple and specific colorimetric detection of Staphylococcus using its volatile 2-[3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl] propanoic acid in the liquid phase and head space of cultures.

    PubMed

    Saranya, Raju; Aarthi, Raju; Sankaran, Krishnan

    2015-05-01

    Spread of drug-resistant Staphylococcus spp. into communities pose danger demanding effective non-invasive and non-destructive tools for its early detection and surveillance. Characteristic volatile organic compounds (VOCs) produced by bacteria offer new diagnostic targets and novel approaches not exploited so far in infectious disease diagnostics. Our search for such characteristic VOC for Staphylococcus spp. led to the depiction of 2-[3-acetoxy-4,4,14-trimethylandrost-8-en-17-yl] propanoic acid (ATMAP), a moderately volatile compound detected both in the culture and headspace when the organism was grown in tryptone soya broth (TSB) medium. A simple and inexpensive colorimetric method (colour change from yellow to orange) using methyl red as the pH indicator provided an absolutely specific way for identifying Staphylococcus spp., The assay performed in liquid cultures (7-h growth in TSB) as well as in the headspace of plate cultures (grown for 10 h on TSA) was optimised in a 96-well plate and 12-well plate formats, respectively, employing a set of positive and negative strains. Only Staphylococcus spp. showed the distinct colour change from yellow to orange due to the production of the above VOC while in the case of other organisms, the reagent remained yellow. The method validated using known clinical and environmental strains (56 including Staphylococcus, Proteus, Pseudomonas, Klebsiella, Bacillus, Shigella and Escherichia coli) was found to be highly efficient showing 100% specificity and sensitivity. Such simple methods of bacterial pathogen identification are expected to form the next generation tools for the control of infectious diseases through early detection and surveillance of causative agents. PMID:25900191

  10. Hypervelocity plate acceleration

    SciTech Connect

    Marsh, S.P.; Tan, T.H.

    1991-01-01

    Shock tubes have been used to accelerate 1.5-mm-thick stainless steel plates to high velocity while retaining their integrity. The fast shock tubes are 5.1-cm-diameter, 15.2-cm-long cylinders of PBX-9501 explosive containing a 1.1-cm-diameter cylindrical core of low-density polystyrene foam. The plates have been placed directly in contact with one face of the explosive system. Plane-wave detonation was initiated on the opposite face. A Mach disk was formed in the imploding styrofoam core, which provided the impulse required to accelerate the metal plate to high velocity. Parametric studies were made on this system to find the effect of varying plate metal, plate thickness, foam properties, and addition of a barrel. A maximum plate velocity of 9.0 km/s has been observed. 6 refs., 17 figs.

  11. WFPDB: European Plate Archives

    NASA Astrophysics Data System (ADS)

    Tsvetkov, Milcho

    2007-08-01

    The Wide-Field Plate Database (WFPDB) gives an inventory of all wide-field (>~ 1 sq. deg) photographic observations archived in astronomical institutions over the world. So it facilitates and stimulates their use and preservation as a valuable source of information for future investigations in astronomy. At present WFPDB manages plate-index information for 25% of all existing plates providing on-line access from Sofia (http://www.skyarchive.org/search) and in CDS, Strasbourg. Here we present the new development of WFPDB as an instrument for searching of long term brightness variations of different sky objects stressing on the European photographic plate collections (from existing 2 million wide-field plates more than 55% are in Europe: Germany, Russia, Ukraine, Italy, Czech Republic, etc.). We comment examples of digitization (with flatbed scanners) of the European plate archives in Sonneberg, Pulkovo, Asiago, Byurakan, Bamberg, etc. and virtual links of WFPDB with European AVO, ADS, IBVS.

  12. GOLD PLATING PROCESS

    DOEpatents

    Seegmiller, R.

    1957-08-01

    An improved bath is reported for plating gold on other metals. The composition of the plating bath is as follows: Gold cyanide from about 15 to about 50 grams, potassium cyanide from about 70 to about 125 grams, and sulfonated castor oil from about 0.1 to about 10 cc. The gold plate produced from this bath is smooth, semi-hard, and nonporous.

  13. 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.

  14. CALUTRON FACE PLATE

    DOEpatents

    Brobeck, W.M.

    1959-08-25

    The construction of a removable cover plate for a calutron tank is described. The plate is fabricated of a rectangular frame member to which is welded a bowed or dished plate of thin steel, reinforced with transverse stiffening ribs. When the tank is placed between the poles of a magnet, the plate may be pivoted away from the tank and magnet and is adapted to support the ion separation mechanism secured to its inner side as well as the vacuum load within the tank.

  15. PLATES WITH OXIDE INSERTS

    DOEpatents

    West, J.M.; Schumar, J.F.

    1958-06-10

    Planar-type fuel assemblies for nuclear reactors are described, particularly those comprising fuel in the oxide form such as thoria and urania. The fuel assembly consists of a plurality of parallel spaced fuel plate mennbers having their longitudinal side edges attached to two parallel supporting side plates, thereby providing coolant flow channels between the opposite faces of adjacent fuel plates. The fuel plates are comprised of a plurality of longitudinally extending tubular sections connected by web portions, the tubular sections being filled with a plurality of pellets of the fuel material and the pellets being thermally bonded to the inside of the tubular section by lead.

  16. Alteration of the aPA ELISA by UV exposure of polystyrene microtiter plates.

    PubMed

    Goldberg, J S; Wagenknecht, D R; McIntyre, J A

    1996-01-01

    Interlaboratory inconsistencies in antiphospholipid antibody (aPA) solid phase assays have prompted controversy in clinical laboratory testing for aPA. We found that the aPA ELISA can be influenced by the type of microtiter plate utilized and by the conditions in which the plates are stored. By exposing 96-well, flat-bottom polystyrene microtiter plates to short wave UV light (254 nm), the aPA ELISA signal decreased in a UV dose-dependent manner. No effect was seen with long wave UV light (366 nm). These results were independent of the antibody isotype under study or the phospholipid (PL) antigen used: anionic phosphatidylserine (PS) and cardiolipin (CL), or zwitterionic phosphatidylethanolamine (PE). Purified human beta 2-glycoprotein I (beta 2 GPI), a known cofactor for anionic PL, and rabbit anti-beta 2 GPI antisera were used to demonstrate that beta 2 GPI bound equally to UV treated and untreated microtiter plates. In contrast, recognition of beta 2 GPI on an anionic PL surface was decreased on UV treated plates, suggesting that UV exposure alters the lipid binding properties of the microliter plate. To determine whether UV exposure inhibited PL binding directly or caused a change in the way the PL was bound, the amount of PL bound to UV treated and untreated plates was measured by using fluorescent labeled PS and a fluorimeter. PS binding was decreased by 53% in UV treated wells as compared to untreated wells. These data show that short wave UV exposure reduces PL binding to polystyrene microtiter plates, thereby reducing the amount of beta 2 GPI bound to PL coated ELISA plates. Thus by using UV exposed microtiter plates, decreased or false-negative a PA ELISA results may be obtained for aPA positive plasmas. PMID:8887002

  17. A membrane-based purification process for cell culture-derived influenza A virus.

    PubMed

    Weigel, Thomas; Solomaier, Thomas; Wehmeyer, Sebastian; Peuker, Alessa; Wolff, Michael W; Reichl, Udo

    2016-02-20

    A simple membrane-based purification process for cell culture-derived influenza virus was established that relies on only two chromatographic unit operations to achieve the contamination limits required according to regulatory authorities. After clarification and concentration, a pseudo-affinity membrane adsorber (sulfated cellulose, SCMA) was applied for virus capture. The subsequent polishing step consisted of a salt-tolerant anion exchange membrane adsorber (STMA) to bind residual DNA. For the presented process neither a buffer exchange step nor a nuclease step for further DNA digestion were required. As a starting point, a two-salt strategy (including a polyvalent ion) was employed to screen STMA conditions in a 96-well plate format. After optimization on chromatographic laboratory scale, the virus recovery was up to 97% with a residual DNA level below 0.82%. In addition, the STMA was characterized regarding its dynamic binding capacity and the impact of flow rate on yields and contamination levels. Overall, the total virus yield for influenza virus A/PR/8/34 (H1/N1) of this two-step membrane process was 75%, while the protein and the DNA contamination level could be reduced to 24% and at least 0.5%, respectively. With 19.8μg protein and 1.2ng DNA per monovalent dose, this purity level complies with the limits of the European Pharmacopeia for cell culture-derived vaccines for human use. Overall, the presented downstream process might serve as a generic and economic platform technology for production of cell culture-derived viruses and viral vectors. PMID:26712479

  18. Blue Willow Story Plates

    ERIC Educational Resources Information Center

    Fontes, Kris

    2009-01-01

    In the December 1997 issue of "SchoolArts" is a lesson titled "Blue Willow Story Plates" by Susan Striker. In this article, the author shares how she used this lesson with her middle-school students many times over the years. Here, she describes a Blue Willow plate painting project that her students made.

  19. Earthquakes and plate tectonics

    USGS Publications Warehouse

    Spall, H.

    1977-01-01

    An explanation is to be found in plate tectonics, a concept which has revolutionized thinking in the Earth sciences in the last 10 years. The theory of plate tectonics combines many of the ideas about continental drift (originally proposed in 1912 by Alfred Wegener in Germany) and sea-floor spreading (suggested originally by Harry Hess of Princeton University). 

  20. Turbine vane plate assembly

    DOEpatents

    Schiavo Jr., Anthony L.

    2006-01-10

    A turbine vane assembly includes a turbine vane having first and second shrouds with an elongated airfoil extending between. Each end of the airfoil transitions into a shroud at a respective junction. Each of the shrouds has a plurality of cooling passages, and the airfoil has a plurality of cooling passages extending between the first and second shrouds. A substantially flat inner plate and an outer plate are coupled to each of the first and second shrouds so as to form inner and outer plenums. Each inner plenum is defined between at least the junction and the substantially flat inner plate; each outer plenum is defined between at least the substantially flat inner plate and the outer plate. Each inner plenum is in fluid communication with a respective outer plenum through at least one of the cooling passages in the respective shroud.

  1. Comparison of Assurance GDS(®) MPX ID for Top STEC with Reference Culture Methods for the Detection of E. coli Top 6 STEC; Direct Confirmation of Top 6 STEC from Isolation Plates and Determination of Equivalence of PickPen(®) and FSIS OctoMACS™ Concentration Protocols.

    PubMed

    Feldsine, Philip; Lienau, Andrew H; Shah, Khyati; Immermann, Amy; Soliven, Khanh; Kaur, Mandeep; Kerr, David E; Jucker, Markus; Hammack, Tom; Brodsky, Michael; Agin, James

    2016-01-01

    Assurance GDS(®) MPX ID for Top Shiga toxin-producing Escherichia coli (STEC; MPX ID) was validated according to the AOAC INTERNATIONAL Methods Committee Guidelines for Validation of Microbiological Methods for Foods and Environmental Surfaces as (1) a secondary screening method for specific detection of the Top 6 STEC serogroups (O26, O45, O103, O111, O121, and O145) in raw beef trim, raw ground beef, raw spinach, and on stainless steel; and (2) as a confirmatory method for the identification of pure culture isolates as Top 6 STEC. MPX ID is used in conjunction with the upfront BCS Assurance GDS MPX Top 7 STEC assay. This Performance Tested Method(SM) validation has two main parts: Method Developer studies and the Independent Laboratory study. A total of 180 samples and controls were analyzed. Results showed that MPX ID had no statistically significant differences with the reference culture methods for the detection of Top 6 STEC in the food matrixes (raw beef trim, raw ground beef, and raw spinach) and environmental sponges (stainless steel) studied. Inclusivity/exclusivity studies were also conducted. One hundred percent inclusivity among the 50 Top 6 STEC serovars tested and 100% exclusivity for the 30 non-Top 6 STEC organisms tested were demonstrated. For validation of MPX ID as a confirmatory method for isolated colonies, all inclusivity and exclusivity organisms were streaked for isolation onto five STEC plating media: modified rainbow agar, Levine's eosin-methylene blue (L-EMB) agar, rainbow agar with novobiocin and cefixime, and enterohemolysin agar with selective agents as well as trypticase soy agar with yeast extract. These isolated colonies were suspended and analyzed by Assurance GDS MPX Top 7 STEC and MPX ID. MPX ID was able to correctly confirm all inclusivity organisms from all plate types, except two STEC isolates from L-EMB agar plates only in the Independent Laboratory study. All exclusivity organisms were correctly determined by MPX ID as non

  2. Earthquakes and plate tectonics.

    USGS Publications Warehouse

    Spall, H.

    1982-01-01

    Earthquakes occur at the following three kinds of plate boundary: ocean ridges where the plates are pulled apart, margins where the plates scrape past one another, and margins where one plate is thrust under the other. Thus, we can predict the general regions on the earth's surface where we can expect large earthquakes in the future. We know that each year about 140 earthquakes of magnitude 6 or greater will occur within this area which is 10% of the earth's surface. But on a worldwide basis we cannot say with much accuracy when these events will occur. The reason is that the processes in plate tectonics have been going on for millions of years. Averaged over this interval, plate motions amount to several mm per year. But at any instant in geologic time, for example the year 1982, we do not know, exactly where we are in the worldwide cycle of strain build-up and strain release. Only by monitoring the stress and strain in small areas, for instance, the San Andreas fault, in great detail can we hope to predict when renewed activity in that part of the plate tectonics arena is likely to take place. -from Author

  3. Lohse's historic plate archive

    NASA Astrophysics Data System (ADS)

    Tsvetkov, M.; Tsvetkova, K.; Richter, G.; Scholz, G.; Böhm, P.

    The description and the analysis of Oswald Lohse's astrophotographic plates, collected at the Astrophysical Observatory Potsdam in the period 1879 - 1889, are presented. 67 plates of the archive, taken with the greatest instrument of the observatory at that time - the refractor (D = 0.30 m, F = 5.40 m, scale = 38''/mm) and with the second heliographic objective (D = 0.13 m, F = 1.36 m, scale = 152''/mm) - - survived two world wars in relative good condition. The plate emulsions are from different manufacturers in the beginning of astrophotography (Gädicke, Schleussner, Beernaert, etc.). The sizes of the plates are usually 9x12 cm2, which corresponds to fields of 1.2deg and 5deg respectively for each instrument mentioned above. The average limiting magnitude is 13.0(pg). Besides of the plates received for technical experiments (work on photographic processes, testing of new instruments and methods of observations), the scientific observations follow programs for studies of planet surfaces, bright stars, some double stars, stellar clusters and nebulous objects. Lohse's archive is included into the Wide Field Plate Database (http://www.skyarchive.org) as the oldest systematic one, covering the fields of Orion (M42/43), Pleiades, h & chi Persei, M37, M3, M11, M13, M92, M31, etc. With the PDS 2020 GM+ microdensitometer of Münster University 10 archive plates were digitized.

  4. Cavity-Enhanced Immunoassay Measurements in Microtiter Plates Using BBCEAS.

    PubMed

    Bajuszova, Zuzana; Ali, Zulfiqur; Scott, Simon; Seetohul, L Nitin; Islam, Meez

    2016-05-17

    We report on the first detailed use of broadband cavity enhanced absorption spectroscopy (BBCEAS) as a detection system for immunoassay. A vertical R ≥ 0.99 optical cavity was integrated with a motorized XY stage, which functioned as a receptacle for 96-well microtiter plates. The custom-built cavity enhanced microplate reader was used to make measurements on a commercially available osteocalcin sandwich ELISA kit. A 30-fold increase in path length was obtained with a minimum detectable change in the absorption coefficient, αmin(t), of 5.3 × 10(-5) cm(-1) Hz(-1/2). This corresponded to a 39-fold increase in the sensitivity of measurement when directly compared to measurements in a conventional microplate reader. Separate measurements of a standard STREP-HRP colorimetric reaction in microtiter plates of differing optical quality produced an increase in sensitivity of up to 115-fold compared to a conventional microplate reader. The sensitivity of the developed setup compared favorably with previous liquid-phase cavity enhanced studies and approaches the sensitivity of typical fluorometric ELISAs. It could benefit any biochemical test which uses single pass absorption as a detection method, through either the label free detection of biologically important molecules at lower concentrations or the reduction in the amount of expensive biochemicals needed for a particular test, leading to cheaper tests. PMID:27089516

  5. Tectonic Plate Movement.

    ERIC Educational Resources Information Center

    Landalf, Helen

    1998-01-01

    Presents an activity that employs movement to enable students to understand concepts related to plate tectonics. Argues that movement brings topics to life in a concrete way and helps children retain knowledge. (DDR)

  6. Flat plate solar oven

    SciTech Connect

    Parikh, M.

    1981-01-01

    The construction of an Indian Rs. 186 (US $20.33) flat-plate solar oven is described. Detailed drawings are provided and relevant information on cooking times and temperature for different foods is given.

  7. Plate tectonics: Metamorphic myth

    NASA Astrophysics Data System (ADS)

    Korenaga, Jun

    2016-01-01

    Clear evidence for subduction-induced metamorphism, and thus the operation of plate tectonics on the ancient Earth has been lacking. Theoretical calculations indicate that we may have been looking for something that cannot exist.

  8. Violin plate modes.

    PubMed

    Gough, Colin

    2015-01-01

    As the first step toward developing a generic model for the acoustically radiating vibrational modes of the violin and related instruments, the modes of both freely supported and edge-constrained top and back plates have been investigated as functions of shape, arching height, elastic anisotropy, the f-holes and associated island area, thickness graduations, and the additional boundary constraints of the ribs, soundpost, and bass-bar present in the assembled instrument. Comsol shell structure finite element software has been used as a quasi-experimental tool, with physical and geometric properties varied smoothly, often over several orders of magnitude, allowing the development of the plate modes to be followed continuously from those of an initially square plate to those of doubly-arched, guitar-shaped, orthotropic plates and their dependence on all the above factors. PMID:25618046

  9. Positive battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John R. (Inventor)

    1985-01-01

    The power characteristics of a lead acid battery are improved by incorporating a dispersion of 1 to 10% by weight of a thermodynamically stable conductivity additive, such as conductive tin oxide coated glass fibers (34) of filamentary glass wool (42) in the positive active layer (32) carried on the grid (30) of the positive plate (16). Positive plate potential must be kept high enough to prevent reduction of the tin oxide to tin by utilizing an oversized, precharged positive paste.

  10. Fractal multifiber microchannel plates

    NASA Technical Reports Server (NTRS)

    Cook, Lee M.; Feller, W. B.; Kenter, Almus T.; Chappell, Jon H.

    1992-01-01

    The construction and performance of microchannel plates (MCPs) made using fractal tiling mehtods are reviewed. MCPs with 40 mm active areas having near-perfect channel ordering were produced. These plates demonstrated electrical performance characteristics equivalent to conventionally constructed MCPs. These apparently are the first MCPs which have a sufficiently high degree of order to permit single channel addressability. Potential applications for these devices and the prospects for further development are discussed.

  11. Phononic plate waves.

    PubMed

    Wu, Tsung-Tsong; Hsu, Jin-Chen; Sun, Jia-Hong

    2011-10-01

    In the past two decades, phononic crystals (PCs) which consist of periodically arranged media have attracted considerable interest because of the existence of complete frequency band gaps and maneuverable band structures. Recently, Lamb waves in thin plates with PC structures have started to receive increasing attention for their potential applications in filters, resonators, and waveguides. This paper presents a review of recent works related to phononic plate waves which have recently been published by the authors and coworkers. Theoretical and experimental studies of Lamb waves in 2-D PC plate structures are covered. On the theoretical side, analyses of Lamb waves in 2-D PC plates using the plane wave expansion (PWE) method, finite-difference time-domain (FDTD) method, and finite-element (FE) method are addressed. These methods were applied to study the complete band gaps of Lamb waves, characteristics of the propagating and localized wave modes, and behavior of anomalous refraction, called negative refraction, in the PC plates. The theoretical analyses demonstrated the effects of PC-based negative refraction, lens, waveguides, and resonant cavities. We also discuss the influences of geometrical parameters on the guiding and resonance efficiency and on the frequencies of waveguide and cavity modes. On the experimental side, the design and fabrication of a silicon-based Lamb wave resonator which utilizes PC plates as reflective gratings to form the resonant cavity are discussed. The measured results showed significant improvement of the insertion losses and quality factors of the resonators when the PCs were applied. PMID:21989878

  12. A Rapid and Sensitive Method for Measuring N-Acetylglucosaminidase Activity in Cultured Cells

    PubMed Central

    Mauri, Victor; Lotfi, Parisa; Segatori, Laura; Sardiello, Marco

    2013-01-01

    A rapid and sensitive method to quantitatively assess N-acetylglucosaminidase (NAG) activity in cultured cells is highly desirable for both basic research and clinical studies. NAG activity is deficient in cells from patients with Mucopolysaccharidosis type IIIB (MPS IIIB) due to mutations in NAGLU, the gene that encodes NAG. Currently available techniques for measuring NAG activity in patient-derived cell lines include chromogenic and fluorogenic assays and provide a biochemical method for the diagnosis of MPS IIIB. However, standard protocols require large amounts of cells, cell disruption by sonication or freeze-thawing, and normalization to the cellular protein content, resulting in an error-prone procedure that is material- and time-consuming and that produces highly variable results. Here we report a new procedure for measuring NAG activity in cultured cells. This procedure is based on the use of the fluorogenic NAG substrate, 4-Methylumbelliferyl-2-acetamido-2-deoxy-alpha-D-glucopyranoside (MUG), in a one-step cell assay that does not require cell disruption or post-assay normalization and that employs a low number of cells in 96-well plate format. We show that the NAG one-step cell assay greatly discriminates between wild-type and MPS IIIB patient-derived fibroblasts, thus providing a rapid method for the detection of deficiencies in NAG activity. We also show that the assay is sensitive to changes in NAG activity due to increases in NAGLU expression achieved by either overexpressing the transcription factor EB (TFEB), a master regulator of lysosomal function, or by inducing TFEB activation chemically. Because of its small format, rapidity, sensitivity and reproducibility, the NAG one-step cell assay is suitable for multiple procedures, including the high-throughput screening of chemical libraries to identify modulators of NAG expression, folding and activity, and the investigation of candidate molecules and constructs for applications in enzyme replacement

  13. Prediction of Drug-Induced Liver Injury in Micropatterned Co-cultures Containing iPSC-Derived Human Hepatocytes.

    PubMed

    Ware, Brenton R; Berger, Dustin R; Khetani, Salman R

    2015-06-01

    Primary human hepatocytes (PHHs) are a limited resource for drug screening, their quality for in vitro use can vary considerably across different lots, and a lack of available donor diversity restricts our understanding of how human genetics affect drug-induced liver injury (DILI). Induced pluripotent stem cell-derived human hepatocyte-like cells (iPSC-HHs) could provide a complementary tool to PHHs for high-throughput drug screening, and ultimately enable personalized medicine. Here, we hypothesized that previously developed iPSC-HH-based micropatterned co-cultures (iMPCCs) with murine embryonic fibroblasts could be amenable to long-term drug toxicity assessment. iMPCCs, created in industry-standard 96-well plates, were treated for 6 days with a set of 47 drugs, and multiple functional endpoints (albumin, urea, ATP) were evaluated in dosed cultures against vehicle-only controls to enable binary toxicity decisions. We found that iMPCCs correctly classified 24 of 37 hepatotoxic drugs (65% sensitivity), while all 10 non-toxic drugs tested were classified as such in iMPCCs (100% specificity). On the other hand, conventional confluent cultures of iPSC-HHs failed to detect several liver toxins that were picked up in iMPCCs. Results for DILI detection in iMPCCs were remarkably similar to published data in PHH-MPCCs (65% versus 70% sensitivity) that were dosed with the same drugs. Furthermore, iMPCCs detected the relative hepatotoxicity of structural drug analogs and recapitulated known mechanisms of acetaminophen toxicity in vitro. In conclusion, iMPCCs could provide a robust tool to screen for DILI potential of large compound libraries in early stages of drug development using an abundant supply of commercially available iPSC-HHs. PMID:25716675

  14. Caribbean plate interactions

    SciTech Connect

    Ball, M. )

    1993-02-01

    Vector analysis of plate motions, derived from studies of Atlantic magnetic lineations and fracture zone trends, indicates the following relative movements between the Caribbean, North American, and South American Plates. (1) During Early Jurassic to Early Cretaceous, the North American Plate moved 1900 km westward and 900 km northward relative to the South American Plate. A broad zone including the Caribbean region, i.e., the zone between the North and South America Plates, was a site of left-lateral shear and north-south extension. (2) During Early Cretaceous to Late Cretaceous, the North American Mate moved an additional 1200 km westward relative to South America across this zone. (3) During Late Cretaceous to the end of the Eocene, the North American Plate moved 200 km westward and 400 km northward relative to the South American Plate. (4) From the end of the Eocene to near the end of the Miocene, North America converged on South America some 200 km and moved 100 km eastward relative to it. Through the Mesozoic and earliest Tertiary history of the Caribbean, the region was a shear zone within which left-lateral displacement exceeded 3000 km and north-south extension exceeded 1300 km. In regard to time, 80% of the history of the Caribbean region is one of north-south extension and left-lateral shear. In terms of space, 97% of the shear is left-lateral and the ratio of divergence versus convergence is 7 to 1. Thus, characterizing the Caribbean region, and the Atlantic to its east, as a zone of north-south extension and left-lateral shear, is a fair generalization.

  15. Using Plate Mapping to Examine Portion Size and Plate Composition for Large and Small Divided Plates

    PubMed Central

    Sharp, David E.; Sobal, Jeffery; Wansink, Brian

    2015-01-01

    Does the size of a plate influence the serving of all items equally, or does it influence the serving of some foods – such as meat versus starch versus vegetables – very differently? To examine this, we utilize a promising new plate mapping method where people drew a meal on a paper plate to examine sensitivity to small versus large three-compartment divided plates in portion size and meal composition in a sample of 109 university students. The total drawn meal area was 37% bigger on large plates than small plates, that is, the portion of plate coverage did not differ by plate size. Men and women drew bigger vegetable portions and men drew bigger meat portions on large plates when compared to small plates. These results suggest that men and women are differentially sensitive to plate size for overall meal size and for meal composition. Implications for decreasing portion size and improving meal balance are discussed. PMID:25280373

  16. Naturally Derived Iron Oxide Nanowires from Bacteria for Magnetically Triggered Drug Release and Cancer Hyperthermia in 2D and 3D Culture Environments: Bacteria Biofilm to Potent Cancer Therapeutic.

    PubMed

    Kumeria, Tushar; Maher, Shaheer; Wang, Ye; Kaur, Gagandeep; Wang, Luoshan; Erkelens, Mason; Forward, Peter; Lambert, Martin F; Evdokiou, Andreas; Losic, Dusan

    2016-08-01

    Iron oxide nanowires produced by bacteria (Mariprofundus ferrooxydans) are demonstrated as new multifunctional drug carriers for triggered therapeutics release and cancer hyperthmia applications. Iron oxide nanowires are obtained from biofilm waste in the bore system used to pump saline groundwater into the River Murray, South Australia (Australia) and processed into individual nanowires with extensive magnetic properties. The drug carrier capabilities of these iron oxide nanowires (Bac-FeOxNWs) are assessed by loading anticancer drug (doxorubicin, Dox) followed by measuring its elution under sustained and triggered release conditions using alternating magnetic field (AMF). The cytotoxicity of Bac-FeOxNWs assessed in 2D (96 well plate) and 3D (Matrigel) cell cultures using MDA-MB231-TXSA human breast cancer cells and mouse RAW 264.7 macrophage cells shows that these Bac-FeOxNWs are biocompatible even at concentrations as high as 250 μg/mL after 24 h of incubation. Finally, we demonstrate the capabilities of Bac-FeOxNWs as potential hyperthermia agent in 3D culture setup. Application of AMF increased the local temperature by 14 °C resulting in approximately 34% decrease in cell viability. Our results demonstrate that these naturally produced nanowires in the form of biofilm can efficiently act as drug carriers with triggered payload release and magnetothermal heating features for potential anticancer therapeutics applications. PMID:27428076

  17. Cadmium plating replacements

    SciTech Connect

    Nelson, M.J.; Groshart, E.C.

    1995-03-01

    The Boeing Company has been searching for replacements to cadmium plate. Two alloy plating systems seem close to meeting the needs of a cadmium replacement. The two alloys, zinc-nickel and tin-zinc are from alloy plating baths; both baths are neutral pH. The alloys meet the requirements for salt fog corrosion resistance, and both alloys excel as a paint base. Currently, tests are being performed on standard fasteners to compare zinc-nickel and tin-zinc on threaded hardware where cadmium is heavily used. The Hydrogen embrittlement propensity of the zinc-nickel bath has been tested, and just beginning for the tin-zinc bath. Another area of interest is the electrical properties on aluminum for tin-zinc and will be discussed. The zinc-nickel alloy plating bath is in production in Boeing Commercial Airplane Group for non-critical low strength steels. The outlook is promising that these two coatings will help The Boeing Company significantly reduce its dependence on cadmium plating.

  18. Cadmium plating replacements

    NASA Technical Reports Server (NTRS)

    Nelson, Mary J.; Groshart, Earl C.

    1995-01-01

    The Boeing Company has been searching for replacements to cadmium plate. Two alloy plating systems seem close to meeting the needs of a cadmium replacement. The two alloys, zinc-nickel and tin-zinc are from alloy plating baths; both baths are neutral pH. The alloys meet the requirements for salt fog corrosion resistance, and both alloys excel as a paint base. Currently, tests are being performed on standard fasteners to compare zinc-nickel and tin-zinc on threaded hardware where cadmium is heavily used. The Hydrogen embrittlement propensity of the zinc-nickel bath has been tested, and just beginning for the tin-zinc bath. Another area of interest is the electrical properties on aluminum for tin-zinc and will be discussed. The zinc-nickel alloy plating bath is in production in Boeing Commercial Airplane Group for non-critical low strength steels. The outlook is promising that these two coatings will help The Boeing Company significantly reduce its dependence on cadmium plating.

  19. Insulin-like growth factor-I (IGF-I) protects cultured equine Leydig cells from undergoing apoptosis.

    PubMed

    Yoon, M J; Roser, J F

    2010-12-01

    Leydig cells located in the interstitial space of the testicular parenchyma produce testosterone which plays a critical role in the maintenance and restoration of spermatogenesis in many species, including horses. For normal spermatogenesis, maintaining Leydig cells is critical to provide an optimal and constant level of testosterone. Recently, an anti-apoptotic effect of IGF-I in testicular cells in rats has been reported, but a similar effect of IGF-I on equine Leydig cells remains to be elucidated. If IGF-I also protects stallion testicular cells from undergoing apoptosis, then IGF-I may have potential as a treatment regime to prevent testicular degeneration. The present study was designed to evaluate the anti-apoptotic effect of IGF-I on cultured equine Leydig cells. Testes were collected from 5 post-pubertal stallions (2-4 years old) during routine castrations. A highly purified preparation of equine Leydig cells was obtained from a discontinuous Percoll gradient. Purity of equine Leydig cells was assessed using histochemical 3β-HSD staining. Equine Leydig cells and selected doses of recombinant human IGF-1 (rhIGF-I; Parlow A.F., National Hormone and Peptide Program, Harbor-UCLA Medical Center) were added to wells of 24 or 96 well culture plates in triplicate and cultured for 24 or 48 h under 95% air:5% CO(2) at 34°C. After 24 or 48 h incubation, apoptotic rate was assessed using a Cell Death Detection ELISA kit. Significantly lower apoptotic rates were observed in equine Leydig cells cultured with 5, 10, or 50ng/ml of rhIGF-I compared with control cells cultured without rhIGF-I for 24h. Exposure to 1, 5, 10 or 50 ng/ml of rhIGF-I significantly decreased apoptotic rate in equine Leydig cells cultured for 48 h. After 48 h incubation, cells were labeled with Annexin V and propodium iodine to determine the populations of healthy, apoptotic, and necrotic cells by counting stained cells using a Nikon Eclipse inverted fluorescence microscope. As a percentage of

  20. Bipolar battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1987-01-01

    A liquid-impermeable plate (10) having through-plate conductivity with essentially zero resistance comprises an insulator sheet (12) having a series of spaced perforations (14) each of which contains a metal element (16) sealingly received into the perforation (14). A low-cost plate can readily be manufactured by punching a thermoplastic sheet (40) such as polypropylene with a punching tool (52), filling the apertures with led spheres (63) having a diameter smaller than the holes (50) but larger than the thickness of the sheet, sweeping excess spheres (62) off the sheet with a doctor blade (60) and then pressing a heated platen (74) onto the sheet to swage the spheres into a cylindrical shape and melt the surrounding resin to form a liquid-impermeable collar (4) sealing the metal into the sheet.

  1. Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers.

    PubMed

    Grosch, Jan-Hendrik; Sieben, Michaela; Lattermann, Clemens; Kauffmann, Kira; Büchs, Jochen; Spieß, Antje C

    2016-03-01

    Microtiter plates (MTP) and automatized techniques are increasingly applied in the field of biotechnology. However, the susceptibility of MTPs to edge effects such as thermal gradients can lead to high variation of measured enzyme activities. In an effort to enhance experimental reliability, to quantify, and to minimize instrument-caused deviations in enzyme kinetics between two MTP-readers, we comprehensively quantified temperature distribution in 96-well MTPs. We demonstrated the robust application of the absorbance dye cresol red as easily applicable temperature indicator in cuvettes and MTPs and determined its accuracy to ±0.16°C. We then quantified temperature distributions in 96-well MTPs revealing temperature deviations over single MTP of up to 2.2°C and different patterns in two commercial devices (BioTek Synergy 4 and Synergy Mx). The obtained liquid temperature was shown to be substantially controlled by evaporation. The temperature-induced enzyme activity variation within MTPs amounted to about 20 %. Activity deviations between MTPs and to those in cuvettes were determined to 40 % due to deviations from the set temperature in MTPs. In conclusion, we propose a better control of experimental conditions in MTPs or alternative experimental systems for reliable determination of kinetic parameters for bioprocess development. PMID:26709721

  2. Nitrided Metallic Bipolar Plates

    SciTech Connect

    Brady, Michael P; Tortorelli, Peter F; Pihl, Josh A; Toops, Todd J; More, Karren Leslie; Meyer III, Harry M; Vitek, John Michael; Wang, Heli; Turner, John; Wilson, Mahlon; Garzon, Fernando; Rockward, Tommy; Connors, Dan; Rakowski, Jim; Gervasio, Don

    2008-01-01

    The objectives are: (1) Develop and optimize stainless steel alloys amenable to formation of a protective Cr-nitride surface by gas nitridation, at a sufficiently low cost to meet DOE targets and with sufficient ductility to permit manufacture by stamping. (2) Demonstrate capability of nitridation to yield high-quality stainless steel bipolar plates from thin stamped alloy foils (no significant stamped foil warping or embrittlement). (3) Demonstrate single-cell fuel cell performance of stamped and nitrided alloy foils equivalent to that of machined graphite plates of the same flow-field design ({approx}750-1,000 h, cyclic conditions, to include quantification of metal ion contamination of the membrane electrode assembly [MEA] and contact resistance increase attributable to the bipolar plates). (4) Demonstrate potential for adoption in automotive fuel cell stacks. Thin stamped metallic bipolar plates offer the potential for (1) significantly lower cost than currently-used machined graphite bipolar plates, (2) reduced weight/volume, and (3) better performance and amenability to high volume manufacture than developmental polymer/carbon fiber and graphite composite bipolar plates. However, most metals exhibit inadequate corrosion resistance in proton exchange membrane fuel cell (PEMFC) environments. This behavior leads to high electrical resistance due to the formation of surface oxides and/or contamination of the MEA by metallic ions, both of which can significantly degrade fuel cell performance. Metal nitrides offer electrical conductivities up to an order of magnitude greater than that of graphite and are highly corrosion resistant. Unfortunately, most conventional coating methods (for metal nitrides) are too expensive for PEMFC stack commercialization or tend to leave pinhole defects, which result in accelerated local corrosion and unacceptable performance.

  3. Plate-mantle coupling from post-Pangea plate kinematics

    NASA Astrophysics Data System (ADS)

    Zahirovic, Sabin; Dietmar Müller, R.; Seton, Maria; Flament, Nicolas

    2015-04-01

    Convection in the Earth's mantle that involves plates at the surfaces gives rise to plate velocities that vary through time and depend on the balance of plate boundary forces, with the present-day providing a snapshot of this ongoing process. However, present-day plate velocities do not capture plate behaviour over geologically representative timeframes and thus cannot be used to evaluate factors limiting plate velocities. Previous studies investigated the effects of continental keels on plate speeds by either using the present-day snapshot or a limited number of reconstructed plate configurations, often leading to conflicting results. For example, an early assumption was that continental keels (especially cratons) were unlikely to impede fast plate motions because India's velocity approached ~20 cm/yr in the Eocene prior to the collision with Eurasia. We employ a modern plate reconstruction approach with evolving global topological plate boundaries for the post-Pangea timeframe (since 200 Ma) to evaluate factors controlling plate velocities. Plate boundary configurations and plate velocities are extracted from the open-source and cross-platform plate reconstruction package GPlates (www.gplates.org) at 1 Myr intervals. For each plate, at each timestep, the area of continental and cratonic lithosphere is calculated to evaluate the effect on plate velocities. Our results support that oceanic plates tend to be 2-3 times faster than plates with large portion of continental plate area, consistent with predictions of numerical models of mantle convection. The fastest plates (~8.5 cm/yr RMS) are dominated by oceanic plate area and high subducting portion of plate perimeter, while the slowest plates (~2.6-2.8 cm/yr RMS) are dominated by continental plate area and bounded by transforms and mid-oceanic ridge segments. Importantly, increasing cratonic fractions (both Proterozoic and Archean lithosphere) significantly impede plate velocities, suggesting that deep continental

  4. Reduced Plating Ignitron

    NASA Technical Reports Server (NTRS)

    Polzin, Kurt A (Inventor); Pearson, J Boise (Inventor)

    2014-01-01

    An ignitron apparatus has an airtight tubular housing having a first sealed end and a second sealed end. An anode is connected at the first sealed end, projecting into the housing, and a recess at the second sealed and forms a well which contains a quantity of liquid gallium or gallium alloy making up the cathode. An ignitor projects through the liquid metal and into the housing. The inner surface of the housing includes at least one plating-reduction structure to prevent electrical shorting of the apparatus caused by plating of the liquid metal.

  5. NICKEL PLATING PROCESS

    DOEpatents

    Hoover, T.B.; Zava, T.E.

    1959-05-12

    A simplified process is presented for plating nickel by the vapor decomposition of nickel carbonyl. In a preferred form of the invention a solid surface is nickel plated by subjecting the surface to contact with a mixture containing by volume approximately 20% nickel carbonyl vapor, 2% hydrogen sulfide and .l% water vapor or 1% oxygen and the remainder carbon dioxide at room temperature until the desired thickness of nickel is obtained. The advantage of this composition over others is that the normally explosive nickel carbonyl is greatly stabilized.

  6. Optimum Ho:YAG laser irradiation condition on percutaneous laser disk decompression (PLDD): in-vitro quantitive evaluation of influence on disk cells using three-dimensional cell culture system

    NASA Astrophysics Data System (ADS)

    Sato, Masato; Ishihara, Miya; Arai, Tsunenori; Asazuma, Takashi; Kikuchi, Toshiyuki; Kikuchi, Makoto; Fujikawa, Kyosuke

    1999-06-01

    The purpose of this study is to evaluate the influence on disc cells after laser irradiation using three-dimensional (3-D) culture system and to clarify the optimum Ho:YAG laser irradiation condition on percutaneous laser disc decompression (PLDD) therapy. Since the Ho:YAG laser ablation is characterized by water-vapor bubble dynamics with pressure wave, thermal effect on cell metabolism might occur in the intervertebral disc. We studied the disc cell damage on the metabolic point of view to investigate the optimum irradiation parameter of the Ho:YAG laser. We have developed the 3-D cultured disc cell system using agarose gel to investigate laser - disc cell interaction. This culture system provides a highly in vivo-like environment for disc cells in which cell- extracellular matrix interactions appear to be more important than contacts among cells. Intervertebral discs were obtained from Japanese white. The isolated disc cells were seeded in 96-well culture plates at the cell densities of 1 X 106 cells/ml, and incubated for 12 days. A pulsed Ho:YAG laser was delivered through a 200 micrometer-core diameter single silica glass fiber. On the agarose gel including the 3-D cultured disc cells, we used the Ho:YAG laser irradiation energy ranging from 40 to 180 mJ/pulse at the fiber end. Cytotoxicity and matrix synthesis after the laser irradiations were evaluated in time course to determine the optimum condition of laser irradiations. It was confirmed that laser irradiation causes necrosis of the cells and additionally produces apoptosis depending on the condition. The ability of matrix synthesis was maintained even after the irradiation, which differed depending on the irradiation conditions. The optimum irradiation conditions seemed related to the preservation of intact area and the acceleration of matrix synthesis in reactive area.

  7. Nuclear reactor alignment plate configuration

    DOEpatents

    Altman, David A; Forsyth, David R; Smith, Richard E; Singleton, Norman R

    2014-01-28

    An alignment plate that is attached to a core barrel of a pressurized water reactor and fits within slots within a top plate of a lower core shroud and upper core plate to maintain lateral alignment of the reactor internals. The alignment plate is connected to the core barrel through two vertically-spaced dowel pins that extend from the outside surface of the core barrel through a reinforcement pad and into corresponding holes in the alignment plate. Additionally, threaded fasteners are inserted around the perimeter of the reinforcement pad and into the alignment plate to further secure the alignment plate to the core barrel. A fillet weld also is deposited around the perimeter of the reinforcement pad. To accomodate thermal growth between the alignment plate and the core barrel, a gap is left above, below and at both sides of one of the dowel pins in the alignment plate holes through with the dowel pins pass.

  8. Growth Plate Injuries

    MedlinePlus

    ... or crushed, the growth plate may close prematurely, forming a bony bridge or “bar.” The risk of ... this publication: James S. Panagis, M.D., M.P.H., NIAMS/NIH; R. Tracy Ballock, M.D., Case ...

  9. The Plate Tectonics Project

    ERIC Educational Resources Information Center

    Hein, Annamae J.

    2011-01-01

    The Plate Tectonics Project is a multiday, inquiry-based unit that facilitates students as self-motivated learners. Reliable Web sites are offered to assist with lessons, and a summative rubric is used to facilitate the holistic nature of the project. After each topic (parts of the Earth, continental drift, etc.) is covered, the students will…

  10. INL HIP Plate Fabrication

    SciTech Connect

    B. H. Park; C. R. Clark; J. F. Jue

    2010-02-01

    This document outlines the process used to bond monolithic fuel plates by Hot Isostatic Pressing (HIP). This method was developed at Idaho National Laboratory (INL) for the Reduced Enrichment for Research and Test Reactors (RERTR) program. These foils have been used in a number of irradiation experiments in support of the United States Global Threat Reduction Initiative (GTRI) program.

  11. Unitary plate electrode

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor); Clough, Thomas J. (Inventor); Josefowicz, Jack Y. (Inventor); Sibert, John W. (Inventor)

    1985-01-01

    The unitary electrode (10) comprises a porous sheet (12) of fiberglass the strands (14) of which contain a coating (16) of conductive tin oxide. The lower portion of the sheet contains a layer (18) of resin and the upper layer (20) contains lead dioxide forming a positive active electrode on an electrolyte-impervious layer. The strands (14) form a continuous conduction path through both layers (16, 18). Tin oxide is prevented from reduction by coating the surface of the plate facing the negative electrode with a conductive, impervious layer resistant to reduction such as a thin film (130) of lead or graphite filled resin adhered to the plate with a layer (31) of conductive adhesive. The plate (10) can be formed by casting a molten resin from kettle (60) onto a sheet of glass wool (56) overlying a sheet of lead foil and then applying positive active paste from hopper (64) into the upper layer (68). The plate can also be formed by passing an assembly of a sheet ( 80) of resin, a sheet (86) of sintered glass and a sheet (90) of lead between the nip (92) of heated rollers (93, 95) and then filling lead oxide into the pores (116) of the upper layer (118).

  12. Optimizing Two-Color Semiconductor Nanocrystal Immunoassays in Single Well Microtiter Plate Formats

    PubMed Central

    Sapsford, Kim E.; Spindel, Samantha; Jennings, Travis; Tao, Guoliang; Triulzi, Robert C.; Algar, W. Russ; Medintz, Igor L.

    2011-01-01

    The simultaneous detection of two analytes, chicken IgY (IgG) and Staphylococcal enterotoxin B (SEB), in the single well of a 96-well plate is demonstrated using luminescent semiconductor quantum dot nanocrystal (NC) tracers. The NC-labeled antibodies were prepared via sulfhydryl-reactive chemistry using a facile protocol that took <3 h. Dose response curves for each target were evaluated in a single immunoassay format and compared to Cy5, a fluorophore commonly used in fluorescent immunoassays, and found to be equivalent. Immunoassays were then performed in a duplex format, demonstrating multiplex detection in a single well with limits of detection equivalent to the single assay format: 9.8 ng/mL chicken IgG and 7.8 ng/mL SEB. PMID:22164051

  13. Renewable liquid reflecting zone plate

    DOEpatents

    Toor, Arthur; Ryutov, Dmitri D.

    2003-12-09

    A renewable liquid reflecting zone plate. Electrodes are operatively connected to a dielectric liquid in a circular or other arrangement to produce a reflecting zone plate. A system for renewing the liquid uses a penetrable substrate.

  14. MyPlate Food Guide

    MedlinePlus

    ... follow throughout your life. 2. Fruits Like veggies, fruits contain vitamins, minerals, and fiber. The red section of MyPlate is slightly smaller than the green, but together fruits and veggies should fill half your plate. Whole ...

  15. What Are Growth Plate Injuries?

    MedlinePlus

    ... activities. Other reasons for growth plate injuries are:  Child abuse  Injury from extreme cold (for example, frostbite)  Radiation ( ... problems) treats most growth plate injuries. At other times, the child will see a pediatric orthopaedic surgeon (a doctor ...

  16. What Are Growth Plate Injuries?

    MedlinePlus

    ... activities. Other reasons for growth plate injuries are: Child abuse Injury from extreme cold (for example, frostbite) Radiation ( ... problems) treats most growth plate injuries. At other times, the child will see a pediatric orthopaedic surgeon (a doctor ...

  17. Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-Cancer Drug Testing in 3D Culture

    PubMed Central

    Lemmo, Stephanie; Atefi, Ehsan; Luker, Gary D.; Tavana, Hossein

    2014-01-01

    Tumor spheroids are three-dimensional clusters of cancer cells that exhibit characteristics of poorly perfused tumors and hence present a relevant model for testing the efficacy of anti-cancer compounds. The use of spheroids for drug screening is hindered by technological complexities for high throughput generation of consistent size spheroids individually addressable by drug compounds. Here we present and optimize a simple spheroid technology based on the use of an aqueous two-phase system. Cancer cells confined in a drop of the denser aqueous dextran phase are robotically dispensed into a microwell containing the immersion aqueous polyethylene glycol phase. Cells remain within the drop and form a viable spheroid, without a need for any external stimuli. The size of resulting spheroids is sensitive to volume variations of dispensed drops from the air displacement pipetting head of a commercial liquid handling robot. Therefore, we parametrically optimize the process of dispensing of dextran phase drops. For a given cell density, this optimization reproducibly generates consistent size spheroids in standard 96-well plates. In addition, we evaluate the use of a commercial biochemical assay to examine cellular viability of cancer cell spheroids. Spheroids show a dose-dependent response to cisplatin similar to a monolayer culture. However unlike their two-dimensional counterpart, spheroids exhibit resistance to paclitaxel treatment. This technology, which uses only commercially-available reagents and equipment, can potentially expedite anti-cancer drug discovery. Although the use of robotics makes the ATPS spheroid technology particularly useful for drug screening applications, this approach is compatible with simpler liquid handling techniques such as manual micropipetting and offers a straightforward method of 3D cell culture in research laboratories. PMID:25221631

  18. Microchannel plate streak camera

    DOEpatents

    Wang, Ching L.

    1989-01-01

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras. The improved streak camera is far more sensitive to photons (UV to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1000 KeV x-rays.

  19. North American plate dynamics

    NASA Technical Reports Server (NTRS)

    Richardson, Randall M.; Reding, Lynn M.

    1991-01-01

    Deformation within the North American plate in response to various tectonic processes is modeled using an elastic finite element analysis. The tectonic processes considered in the modeling include ridge forces associated with the normal thermal evolution of oceanic lithosphere, shear and normal stresses transmitted across transforms, normal stresses transmitted across convergent boundaries, stresses due to horizontal density contrasts within the continent, and shear tractions applied along the base of the plate. Model stresses are calculated with respect to a lithostatic reference stress state. Shear stresses transmitted across transform boundaries along the San Andreas and Caribbean are small, of the order of 5-10 MPa. Also, compressive stresses of the order of 5-10 MPa transmitted across the major transforms improve the fit to the data. Compressive stresses across convergent margins along the Aleutians and the Middle America trench are important.

  20. Microchannel plate streak camera

    DOEpatents

    Wang, C.L.

    1989-03-21

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras is disclosed. The improved streak camera is far more sensitive to photons (UV to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1,000 KeV x-rays. 3 figs.

  1. Microchannel plate streak camera

    DOEpatents

    Wang, C.L.

    1984-09-28

    An improved streak camera in which a microchannel plate electron multiplier is used in place of or in combination with the photocathode used in prior streak cameras. The improved streak camera is far more sensitive to photons (uv to gamma-rays) than the conventional x-ray streak camera which uses a photocathode. The improved streak camera offers gamma-ray detection with high temporal resolution. It also offers low-energy x-ray detection without attenuation inside the cathode. Using the microchannel plate in the improved camera has resulted in a time resolution of about 150 ps, and has provided a sensitivity sufficient for 1000 keV x-rays.

  2. Bipolar battery plate

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1985-01-01

    A liquid-impermeable plate (10) having throughplate conductivity with essentially zero resistance comprises an insulator sheet (12) having a series of spaced perforations (14) each of which contains a metal element (16) sealingly received into the perforation (14). A low-cost plate can readily be manufactured by punching a thermoplastic sheet (40) such as polypropylene with a punching tool (52), filling the apertures with lead spheres (63) having a diameter smaller than the holes (50) but larger than the thickness of the sheet, sweeping excess spheres (62) off the sheet with a doctor blade (60) and then pressing a heated platen (74) onto the sheet to swage the spheres into a cylindrical shape and melt the surrounding resin to form a liquid-impermeable collar (4) sealing the metal into the sheet.

  3. Plate motion and deformation

    SciTech Connect

    Minster, B.; Prescott, W.; Royden, L.

    1991-02-01

    Our goal is to understand the motions of the plates, the deformation along their boundaries and within their interiors, and the processes that control these tectonic phenomena. In the broadest terms, we must strive to understand the relationships of regional and local deformation to flow in the upper mantle and the rheological, thermal and density structure of the lithosphere. The essential data sets which we require to reach our goal consist of maps of current strain rates at the earth's surface and the distribution of integrated deformation through time as recorded in the geologic record. Our success will depend on the effective synthesis of crustal kinematics with a variety of other geological and geophysical data, within a quantitative theoretical framework describing processes in the earth's interior. Only in this way can we relate the snapshot of current motions and earth structure provided by geodetic and geophysical data with long-term processes operating on the time scales relevant to most geological processes. The wide-spread use of space-based techniques, coupled with traditional geological and geophysical data, promises a revolution in our understanding of the kinematics and dynamics of plate motions over a broad range of spatial and temporal scales and in a variety of geologic settings. The space-based techniques that best address problems in plate motion and deformation are precise space-geodetic positioning -- on land and on the seafloor -- and satellite acquisition of detailed altimetric and remote sensing data in oceanic and continental areas. The overall science objectives for the NASA Solid Earth Science plan for the 1990's, are to Understand the motion and deformation of the lithosphere within and across plate boundaries'', and to understand the dynamics of the mantle, the structure and evolution of the lithosphere, and the landforms that result from local and regional deformation. 57 refs., 7 figs., 2 tabs.

  4. Smov FOS Plate Scale

    NASA Astrophysics Data System (ADS)

    Kinney, Anne

    1994-01-01

    The goal is to measure the precise plate scale and orientation. This will be acheived by performing a raster step and dwell sequence in the 4.3 arcsec aperture. The edges of the aperture should be avoided to prevent vignetting effects. An aperture map is required at each step of the dwell sequence. This test has to be conducted for both the RED and BLUE detectors. We will also determine the offset between the two detectors.

  5. Plated wire memory subsystem

    NASA Technical Reports Server (NTRS)

    Reynolds, L.; Tweed, H.

    1972-01-01

    The work performed entailed the design, development, construction and testing of a 4000 word by 18 bit random access, NDRO plated wire memory for use in conjunction with a spacecraft imput/output unit and central processing unit. The primary design parameters, in order of importance, were high reliability, low power, volume and weight. A single memory unit, referred to as a qualification model, was delivered.

  6. Martian plate tectonics

    NASA Astrophysics Data System (ADS)

    Sleep, N. H.

    1994-03-01

    The northern lowlands of Mars have been produced by plate tectonics. Preexisting old thick highland crust was subducted, while seafloor spreading produced thin lowland crust during late Noachian and Early Hesperian time. In the preferred reconstruction, a breakup margin extended north of Cimmeria Terra between Daedalia Planum and Isidis Planitia where the highland-lowland transition is relatively simple. South dipping subduction occured beneath Arabia Terra and east dipping subduction beneath Tharsis Montes and Tempe Terra. Lineations associated with Gordii Dorsum are attributed to ridge-parallel structures, while Phelegra Montes and Scandia Colles are interpreted as transfer-parallel structures or ridge-fault-fault triple junction tracks. Other than for these few features, there is little topographic roughness in the lowlands. Seafloor spreading, if it occurred, must have been relatively rapid. Quantitative estimates of spreading rate are obtained by considering the physics of seafloor spreading in the lower (approx. 0.4 g) gravity of Mars, the absence of vertical scarps from age differences across fracture zones, and the smooth axial topography. Crustal thickness at a given potential temperature in the mantle source region scales inversely with gravity. Thus, the velocity of the rough-smooth transition for axial topography also scales inversely with gravity. Plate reorganizations where young crust becomes difficult to subduct are another constraint on spreading age. Plate tectonics, if it occurred, dominated the thermal and stress history of the planet. A geochemical implication is that the lower gravity of Mars allows deeper hydrothermal circulation through cracks and hence more hydration of oceanic crust so that more water is easily subducted than on the Earth. Age and structural relationships from photogeology as well as median wavelength gravity anomalies across the now dead breakup and subduction margins are the data most likely to test and modify hypotheses

  7. Elastic plate spallation

    NASA Technical Reports Server (NTRS)

    Oline, L.; Medaglia, J.

    1972-01-01

    The dynamic finite element method was used to investigate elastic stress waves in a plate. Strain displacement and stress strain relations are discussed along with the stiffness and mass matrix. The results of studying point load, and distributed load over small, intermediate, and large radii are reported. The derivation of finite element matrices, and the derivation of lumped and consistent matrices for one dimensional problems with Laplace transfer solutions are included. The computer program JMMSPALL is also included.

  8. Plates with Incompatible Prestrain

    NASA Astrophysics Data System (ADS)

    Bhattacharya, Kaushik; Lewicka, Marta; Schäffner, Mathias

    2016-07-01

    We study effective elastic behavior of the incompatibly prestrained thin plates, where the prestrain is independent of thickness and uniform through the plate's thickness h. We model such plates as three-dimensional elastic bodies with a prescribed pointwise stress-free state characterized by a Riemannian metric G, and seek the limiting behavior as {h to 0}. We first establish that when the energy per volume scales as the second power of h, the resulting {Γ} -limit is a Kirchhoff-type bending theory. We then show the somewhat surprising result that there exist non-immersible metrics G for whom the infimum energy (per volume) scales smaller than h 2. This implies that the minimizing sequence of deformations carries nontrivial residual three-dimensional energy but it has zero bending energy as seen from the limit Kirchhoff theory perspective. Another implication is that other asymptotic scenarios are valid in appropriate smaller scaling regimes of energy. We characterize the metrics G with the above property, showing that the zero bending energy in the Kirchhoff limit occurs if and only if the Riemann curvatures R 1213, R 1223 and R 1212 of G vanish identically. We illustrate our findings with examples; of particular interest is an example where {G_{2 × 2}}, the two-dimensional restriction of G, is flat but the plate still exhibits the energy scaling of the Föppl-von Kármán type. Finally, we apply these results to a model of nematic glass, including a characterization of the condition when the metric is immersible, for {G = Id3 + γ n ⊗ n} given in terms of the inhomogeneous unit director field distribution { n in R^3}.

  9. Perfused Multiwell Plate for 3D Liver Tissue Engineering

    PubMed Central

    Domansky, Karel; Inman, Walker; Serdy, James; Dash, Ajit; Lim, Matthew H. M.

    2014-01-01

    In vitro models that capture the complexity of in vivo tissue and organ behaviors in a scalable and easy-to-use format are desirable for drug discovery. To address this, we have developed a bioreactor that fosters maintenance of 3D tissue cultures under constant perfusion and we have integrated multiple bioreactors into an array in a multiwell plate format. All bioreactors are fluidically isolated from each other. Each bioreactor in the array contains a scaffold that supports formation of hundreds of 3D microscale tissue units. The tissue units are perfused with cell culture medium circulated within the bioreactor by integrated pneumatic diaphragm micropumps. Electronic controls for the pumps are kept outside the incubator and connected to the perfused multiwell by pneumatic lines. The docking design and open-well bioreactor layout make handling perfused multiwell plates similar to using standard multiwell tissue culture plates. A model of oxygen consumption and transport in the circulating culture medium was used to predict appropriate operating parameters for primary liver cultures. Oxygen concentrations at key locations in the system were then measured as a function of flow rate and time after initiation of culture to determine oxygen consumption rates. After seven days in culture, tissue formed from cells seeded in the perfused multiwell reactor remained functionally viable as assessed by immunostaining for hepatocyte and liver sinusoidal endothelial cell (LSEC) phenotypic markers. PMID:20024050

  10. Origin of Small Tectonic Plates

    NASA Astrophysics Data System (ADS)

    Mallard, C.; Coltice, N.; Seton, M.; Müller, D.; Tackley, P.

    2015-12-01

    The plate tectonic theory allowed to split the Earth surface into 6 (Le Pichon 1968) to 52 tectonic plates (Bird 2003). These plates are separated into two groups: the first of 7 large plates and the second of numerous smaller plates (Morra et al 2013). Previous studies using the reconstruction of the past 200 My, suggest that the size of large plates is driven by mantle flow. But the tools employed are descriptive (Morra et al 2013, Sornette and Pisarenko 2003), hence ignoring forces and physical principles within the lithosphere and the mantle. The processes at the origin of small plates remain unknown. We developed a new approach to explain the plate sizes. We demonstrate that the physics of convection drives it. We applied plate tectonics theory on 3D spherical convection models generating plate-like motions, which give access to a complete survey of data: velocities, viscosity and heat flow. Our data show that (1) the large plates depend on the dominating scale of the convective flow due to the initiation or the shutdown of subductions; (2) the smaller plates are generated thanks to large variability of regional stresses along subduction zone by slab pull and suction influenced by the geometry of trenches. Our results are consistent with the quick reorganizations of back-arc basins occuring synchronously with the modification of subduction zones geometry around the Pacific plate (Sdrolias et al 2004). Hence, we conclude that (1) the decreasing number of small plates in the plate reconstructions back in time is an artifact induced by their short lifetime, that is why they are artificially ignored; (2) the geometry of past trenches is simplified leading to an underestimation of the length of subduction zones.

  11. Plate tectonics conserves angular momentum

    NASA Astrophysics Data System (ADS)

    Bowin, C.

    2009-03-01

    A new combined understanding of plate tectonics, Earth internal structure, and the role of impulse in deformation of the Earth's crust is presented. Plate accelerations and decelerations have been revealed by iterative filtering of the quaternion history for the Euler poles that define absolute plate motion history for the past 68 million years, and provide an unprecedented precision for plate angular rotation variations with time at 2-million year intervals. Stage poles represent the angular rotation of a plate's motion between adjacent Euler poles, and from which the maximum velocity vector for a plate can be determined. The consistent maximum velocity variations, in turn, yield consistent estimates of plate accelerations and decelerations. The fact that the Pacific plate was shown to accelerate and decelerate, implied that conservation of plate tectonic angular momentum must be globally conserved, and that is confirmed by the results shown here (total angular momentum ~1.4 E+27 kgm2s-1). Accordingly, if a plate decelerates, other plates must increase their angular momentums to compensate. In addition, the azimuth of the maximum velocity vectors yields clues as to why the "bend" in the Emperor-Hawaiian seamount trend occurred near 46 Myr. This report summarizes processing results for 12 of the 14 major tectonic plates of the Earth (except for the Juan de Fuca and Philippine plates). Plate accelerations support the contention that plate tectonics is a product of torques that most likely are sustained by the sinking of positive density anomalies due to phase changes in subducted gabbroic lithosphere at depth in the upper lower mantle (above 1200 km depth). The tectonic plates are pulled along by the sinking of these positive mass anomalies, rather than moving at near constant velocity on the crests of convection cells driven by rising heat. These results imply that spreading centers are primarily passive reactive features, and fracture zones (and wedge-shaped sites

  12. Symmetries in laminated composite plates

    NASA Technical Reports Server (NTRS)

    Noor, A. K.

    1976-01-01

    The different types of symmetry exhibited by laminated anisotropic fibrous composite plates are identified and contrasted with the symmetries of isotropic and homogeneous orthotropic plates. The effects of variations in the fiber orientation and the stacking sequence of the layers on the symmetries exhibited by composite plates are discussed. Both the linear and geometrically nonlinear responses of the plates are considered. A simple procedure is presented for exploiting the symmetries in the finite element analysis. Examples are given of square, skew and polygonal plates where use of symmetry concepts can significantly reduce the scope and cost of analysis.

  13. Fuel cell end plate structure

    DOEpatents

    Guthrie, Robin J.; Katz, Murray; Schroll, Craig R.

    1991-04-23

    The end plates (16) of a fuel cell stack (12) are formed of a thin membrane. Pressure plates (20) exert compressive load through insulation layers (22, 26) to the membrane. Electrical contact between the end plates (16) and electrodes (50, 58) is maintained without deleterious making and breaking of electrical contacts during thermal transients. The thin end plate (16) under compressive load will not distort with a temperature difference across its thickness. Pressure plate (20) experiences a low thermal transient because it is insulated from the cell. The impact on the end plate of any slight deflection created in the pressure plate by temperature difference is minimized by the resilient pressure pad, in the form of insulation, therebetween.

  14. Thermo-responsive polymer aided spheroid culture in cryogel based platform for high throughput drug screening.

    PubMed

    Sarkar, J; Kumar, A

    2016-04-21

    In high throughput cell culture, the paradigm is now shifting from 2D to 3D systems. However, in 3D cell culture systems, it is important that the cells form spheroids with robust cell-cell interactions. We fabricated poly(N-isopropylacrylamide-co-gelatin) cryogel scaffolds for cell culture and inserted them into open-ended 96-well plates that formed a drainage and leakage protected, easy to handle high throughput platform. This platform was used to screen for the optimal concentration of poly(N-isopropylacrylamide) (pNIPAAm) as an external aid to the formation of cellular spheroids. It was found that hepatic cells (Hep G2) seeded in the presence of 0.03% pNIPAAm formed better hepatic spheroids in terms of morphology (as assessed by microscopic analysis and formation of bile canaliculi-like structures) and functionality by day 5 of culture. An increase of 44.22%, 15.75%, 36.44%, 32.05% and 27.02% was observed in glucose consumption (1.925 mM per day per 10(4) cells), albumin synthesis (164.18 ng per day per 10(4) cells), CYP1A1 (304.92 pg per min per 10(4) cells), CYP2A6 (441.23 nM per min per 10(4) cells) and phase II metabolic activity (386.18 nM per min(-1) per 10(4) cells), respectively, upon using 0.03% pNIPAAm, as compared to the 3D control. The platform was tested with other cells such as breast and lung cancer cells and found to be compatible. The cell spheroids were subjected to drug toxicity screening in cryogel based open-ended platforms. It was observed that the spheroids were more resistant to anticancer drugs, as compared to 2D and 3D controls, with approximately 11%-67% increase in the IC50 values of tamoxifen and paclitaxel. The platform also showed dose dependent and reproducible responses to drugs. PMID:27027476

  15. A titer plate-based polymer microfluidic platform for high throughput nucleic acid purification.

    PubMed

    Park, D S-W; Hupert, M L; Witek, M A; You, B H; Datta, P; Guy, J; Lee, J-B; Soper, S A; Nikitopoulos, D E; Murphy, M C

    2008-02-01

    A 96-well solid-phase reversible immobilization (SPRI) reactor plate was designed to demonstrate functional titer plate-based microfluidic platforms. Nickel, large area mold inserts were fabricated using an SU-8 based, UV-LIGA technique on 150 mm diameter silicon substrates. Prior to UV exposure, the prebaked SU-8 resist was flycut to reduce the total thickness variation to less than 5 mum. Excellent UV lithography results, with highly vertical sidewalls, were obtained in the SU-8 by using an UV filter to remove high absorbance wavelengths below 350 nm. Overplating of nickel in the SU-8 patterns produced high quality, high precision, metal mold inserts, which were used to replicate titer plate-based SPRI reactors using hot embossing of polycarbonate (PC). Optimized molding conditions yielded good feature replication fidelity and feature location integrity over the entire surface area. Thermal fusion bonding of the molded PC chips at 150 degrees C resulted in leak-free sealing, which was verified in leakage tests using a fluorescent dye. The assembled SPRI reactor was used for simple, fast purification of genomic DNA from whole cell lysates of several bacterial species, which was verified by PCR amplification of the purified genomic DNA. PMID:17659445

  16. Optimization of Salmonella Typhi biofilm assay on polypropylene microtiter plates using response surface methodology.

    PubMed

    Ganjali Dashti, M; Abdeshahian, P; Sudesh, K; Phua, K K

    2016-04-01

    The objective of this study was to develop an optimized assay for Salmonella Typhi biofilm that mimics the environment of the gallbladder as an experimental model for chronic typhoid fever. Multi-factorial assays are difficult to optimize using traditional one-factor-at-a-time optimization methods. Response surface methodology (RSM) was used to optimize six key variables involved in S. Typhi biofilm formation on cholesterol-coated polypropylene 96-well microtiter plates. The results showed that bile (1.22%), glucose (2%), cholesterol (0.05%) and potassium chloride (0.25%) were critical factors affecting the amount of biofilm produced, but agitation (275 rpm) and sodium chloride (0.5%) had antagonistic effects on each other. Under these optimum conditions the maximum OD reading for biofilm formation was 3.4 (λ600 nm), and the coefficients of variation for intra-plate and inter-plate assays were 3% (n = 20) and 5% (n = 8), respectively. These results showed that RSM is an effective approach for biofilm assay optimization. PMID:26963754

  17. Wobble plate engine

    SciTech Connect

    Derderian, H.; Pronovost, J.

    1980-07-01

    A description is given of a conventional Stirling cycle engine of the type characterized by an elongated cylinder having a closed end and an open end, means for applying heat to the exterior of the cylinder to heat gas in the interior of the cylinder at the closed end thereof, a displacer piston mounted for a reciprocation within the closed end of the cylinder and a power piston axially aligned with the displacer piston and mounted for coaxial reciprocation between the displacer piston and the open end of the cylinder, the reciprocation of the displacer piston leading the corresponding stroke of the power piston by a predetermined phase angle, the improved means for properly phasing the reciprocation of the two pistons and for converting the reciprocation of the power piston into rotation of an output shaft which is parallel to the axis of reciprocation comprising: a supporting frame; an output shaft mounted on said frame for rotation about a first axis which is parallel to the axis of reciprocation of the two pistons, said shaft having an inclined cylindrical bearing surface the axis of which intersects said first axis at an acute angle; gimbal means pivotally mounted on said supporting frame for oscillation about a second axis perpendicular to and intersecting first axis; a wobble plate pivotally mounted on a third axis within said gimbal means for oscillation with said gimbal means and for further oscillation about said third axis which is mutually perpendicular to and intersecting said first and second axes, said wobble plate being rotatably mounted on said inclined cylindrical bearing surface in a plane which is perpendicular to said axis of said bearing surface whereby said output shaft can rotate about said first axis as said wobble plate oscillates about said second and third axes.

  18. Shuttle plate braiding machine

    NASA Technical Reports Server (NTRS)

    Huey, Jr., Cecil O. (Inventor)

    1994-01-01

    A method and apparatus for moving yarn in a selected pattern to form a braided article. The apparatus includes a segmented grid of stationary support elements and a plurality of shuttles configured to carry yarn. The shuttles are supported for movement on the grid assembly and each shuttle includes a retractable plunger for engaging a reciprocating shuttle plate that moves below the grid assembly. Such engagement at selected times causes the shuttles to move about the grid assembly in a selected pattern to form a braided article of a particular geometry.

  19. Reduced hydrogen cadmium plating

    SciTech Connect

    Hoeller, T.; Ross, L. ); Varma, R. ); Agarwala, V.S. )

    1991-01-01

    This paper demonstrates the advantages of using a periodic reverse pulse plating method, incorporating a fast cathodic pulse which is separated from the subsequent anodic/cathodic pulses by a long rest period in producing silvery cadmium coatings on steel from aqueous fluoroborate electrolyte. Also, the deposition obtained by combination of pulse currents and turbulent electrolyte flow system (forced convection of electrolyte, Re {approximately} 20-25,000) result in a near hydrogen-free electrodeposition of fine- grained cadmium. This is confirmed by the determination of diffusible hydrogen by the electrochemical (Barnach Electrode) method.

  20. Plated wire memory subsystem

    NASA Technical Reports Server (NTRS)

    Carpenter, K. H.

    1974-01-01

    The design, construction, and test history of a 4096 word by 18 bit random access NDRO Plated Wire Memory for use in conjunction with a spacecraft input/output and central processing unit is reported. A technical and functional description is given along with diagrams illustrating layout and systems operation. Test data is shown on the procedures and results of system level and memory stack testing, and hybrid circuit screening. A comparison of the most significant physical and performance characteristics of the memory unit versus the specified requirements is also included.

  1. Ultrafast microchannel plate photomultipliers.

    PubMed

    Kume, H; Koyama, K; Nakatsugawa, K; Suzuki, S; Fatlowitz, D

    1988-03-15

    Performance characteristics of several new types of photomultiplier tubes (PMT) with microchannel plates (MCP) are presented in this paper. They are the MCP-PMT with 6-microm diam channels, MCP-PMT with an S-l photocathode, and MCP-PMT with multi (discrete) anode and gatable MCP-PMT. Important requirements of an optical detector for picosecond lasers, fluorescence measurements, and material analysis are low light detectability, ultrafast time response, and versatile operation including modulation. The basic configuration, characteristics, and practical results of these detectors are described. PMID:20531532

  2. Tectonics of the Easter plate

    NASA Technical Reports Server (NTRS)

    Engeln, J. F.; Stein, S.

    1984-01-01

    A new model for the Easter plate is presented in which rift propagation has resulted in the formation of a rigid plate between the propagating and dying ridges. The distribution of earthquakes, eleven new focal mechanisms, and existing bathymetric and magnetic data are used to describe the tectonics of this area. Both the Easter-Nazca and Easter-Pacific Euler poles are sufficiently close to the Easter plate to cause rapid changes in rates and directions of motion along the boundaries. The east and west boundaries are propagating and dying ridges; the southwest boundary is a slow-spreading ridge and the northern boundary is a complex zone of convergent and transform motion. The Easter plate may reflect the tectonics of rift propagation on a large scale, where rigid plate tectonics requires boundary reorientation. Simple schematic models to illustrate the general features and processes which occur at plates resulting from large-scale rift propagation are used.

  3. Localised Plate Motion on Venus

    NASA Astrophysics Data System (ADS)

    Ghail, R. C.

    1996-03-01

    The volcanic and tectonic features observed in Dali Vinculum, Parga Vinculum and Imdr Regio are concentrated at long, narrow, curvilinear zones, with relatively minor volcanism and tectonism between these zones. These zones, whilst more diffuse than terrestrial plate boundaries, nevertheless define the margins of tectonic plates. In contrast to Earth, however, it appears that venusian plates are neither created nor destroyed by lateral motion. Rather, plates are thinned and intruded at vincula plate boundaries, vertically accreted by small-scale intra-plate (planitia) volcanism and perhaps destroyed by delamination of thickened crust in tesserae and montane regions such as Thetis Regio and Ishtar Terra. The diversity in age both between and within these three areas together with the evidence for infrequent, small scale resurfacing in the planitiae are difficult to reconcile with a non-uniformitarian geological process.

  4. Multiple plate hydrostatic viscous damper

    SciTech Connect

    Ludwig, L.P.

    1981-02-01

    A device for damping radial motion of a rotating shaft is described. The damper comprises a series of spaced plates extending in a radial direction. A hydraulic piston is utilized to place a load in these plates. Each annular plate is provided with a suitable hydrostatic bearing geometry on at least one of its faces. This structure provides a high degree of dampening in a rotor case system of turbomachinery in general. The damper is particularly useful in gas turbine engines.

  5. Multiple plate hydrostatic viscous damper

    NASA Technical Reports Server (NTRS)

    Ludwig, L. P. (Inventor)

    1981-01-01

    A device for damping radial motion of a rotating shaft is described. The damper comprises a series of spaced plates extending in a radial direction. A hydraulic piston is utilized to place a load in these plates. Each annular plate is provided with a suitable hydrostatic bearing geometry on at least one of its faces. This structure provides a high degree of dampening in a rotor case system of turbomachinery in general. The damper is particularly useful in gas turbine engines.

  6. Channel plate for DNA sequencing

    DOEpatents

    Douthart, R.J.; Crowell, S.L.

    1998-01-13

    This invention is a channel plate that facilitates data compaction in DNA sequencing. The channel plate has a length, a width and a thickness, and further has a plurality of channels that are parallel. Each channel has a depth partially through the thickness of the channel plate. Additionally an interface edge permits electrical communication across an interface through a buffer to a deposition membrane surface. 15 figs.

  7. Ion plating for the future

    NASA Technical Reports Server (NTRS)

    Spalvins, T.

    1981-01-01

    The ion plating techniques are classified relative to the instrumental set up, evaporation media, and mode of transport. A distinction is drawn between the low vacuum (plasma) and high vacuum (ion beam) techniques. Ion plating technology is discussed at the fundamental and industrial level. At the fundamental level, the capabilities and limitations of the plasma (evaporant flux) and film characteristics are evaluated. And on the industrial level, the performance and potential uses of ion plated films are discussed.

  8. Channel plate for DNA sequencing

    DOEpatents

    Douthart, Richard J.; Crowell, Shannon L.

    1998-01-01

    This invention is a channel plate that facilitates data compaction in DNA sequencing. The channel plate has a length, a width and a thickness, and further has a plurality of channels that are parallel. Each channel has a depth partially through the thickness of the channel plate. Additionally an interface edge permits electrical communication across an interface through a buffer to a deposition membrane surface.

  9. Carbon-assisted flyer plates

    DOEpatents

    Stahl, David B.; Paisley, Dennis L.

    1994-01-01

    A laser driven flyer plate utilizing an optical fiber connected to a laser. The end of the optical fiber has a layer of carbon and a metal layer deposited onto it. The carbon layer provides the laser induced plasma which is superior to the plasma produced from most metals. The carbon layer plasma is capable of providing a flatter flyer plate, converting more of the laser energy to driving plasma, promoting a higher flyer plate acceleration, and providing a more uniform pulse behind the plate. In another embodiment, the laser is in optical communication with a substrate onto which a layer of carbon and a layer of metal have been deposited.

  10. True Shear Parallel Plate Viscometer

    NASA Technical Reports Server (NTRS)

    Ethridge, Edwin; Kaukler, William

    2010-01-01

    This viscometer (which can also be used as a rheometer) is designed for use with liquids over a large temperature range. The device consists of horizontally disposed, similarly sized, parallel plates with a precisely known gap. The lower plate is driven laterally with a motor to apply shear to the liquid in the gap. The upper plate is freely suspended from a double-arm pendulum with a sufficiently long radius to reduce height variations during the swing to negligible levels. A sensitive load cell measures the shear force applied by the liquid to the upper plate. Viscosity is measured by taking the ratio of shear stress to shear rate.

  11. Glass-bead peen plating

    NASA Technical Reports Server (NTRS)

    Graves, J. R.

    1974-01-01

    Peen plating of aluminum, copper, and nickel powders was investigated. Only aluminum was plated successfully within the range of peen plating conditions studied. Optimum plating conditions for aluminum were found to be: (1) bead/powder mixture containing 25 to 35% powder by weight, (2) peening intensity of 0.007A as measured by Almen strip, and (3) glass impact bead diameter of at least 297 microns (0.0117 inches) for depositing-100 mesh aluminum powder. No extensive cleaning or substrate preparation is required beyond removing loose dirt or heavy oil.

  12. Laser-driven flyer plate

    SciTech Connect

    Paisley, D.L.

    1991-09-10

    Disclosed is an apparatus for producing high velocity flyer plates involving placing a layer of dielectric material between a first metal foil and a second metal foil. With laser irradiation through an optical substrate, the first metal foil forms a plasma in the area of the irradiation, between the substrate and the solid portion of the first metal foil. When the pressure between the substrate and the foil reaches the stress limit of the dielectric, the dielectric will break away and launch the flyer plate out of the second metal foil. The mass of the flyer plate is controlled, as no portion of the flyer plate is transformed into a plasma. 2 figures.

  13. Laser-driven flyer plate

    DOEpatents

    Paisley, Dennis L.

    1991-01-01

    Apparatus for producing high velocity flyer plates involving placing a layer of dielectric material between a first metal foil and a second metal foil. With laser irradiation through an optical substrate, the first metal foil forms a plasma in the area of the irradiation, between the substrate and the solid portion of the first metal foil. When the pressure between the substrate and the foil reaches the stress limit of the dielectric, the dielectric will break away and launch the flyer plate out of the second metal foil. The mass of the flyer plate is controlled, as no portion of the flyer plate is transformed into a plasma.

  14. Scintillator plate calorimetry

    SciTech Connect

    Price, L.E.

    1990-01-01

    Calorimetry using scintillator plates or tiles alternated with sheets of (usually heavy) passive absorber has been proven over multiple generations of collider detectors. Recent detectors including UA1, CDF, and ZEUS have shown good results from such calorimeters. The advantages offered by scintillator calorimetry for the SSC environment, in particular, are speed (<10 nsec), excellent energy resolution, low noise, and ease of achieving compensation and hence linearity. On the negative side of the ledger can be placed the historical sensitivity of plastic scintillators to radiation damage, the possibility of nonuniform response because of light attenuation, and the presence of cracks for light collection via wavelength shifting plastic (traditionally in sheet form). This approach to calorimetry is being investigated for SSC use by a collaboration of Ames Laboratory/Iowa State University, Argonne National Laboratory, Bicron Corporation, Florida State University, Louisiana State University, University of Mississippi, Oak Ridge National Laboratory, Virginia Polytechnic Institute and State University, Westinghouse Electric Corporation, and University of Wisconsin.

  15. An Optimized High Throughput Clean-Up Method Using Mixed-Mode SPE Plate for the Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS

    PubMed Central

    Wang, Wan; Qin, Suzi; Li, Linsen; Chen, Xiaohua; Wang, Qunjie; Wei, Junfu

    2015-01-01

    A high throughput sample preparation method was developed utilizing mixed-mode solid phase extraction (SPE) in 96-well plate format for the determination of free arachidonic acid in plasma by LC-MS/MS. Plasma was mixed with 3% aqueous ammonia and loaded into each well of 96-well plate. After washing with water and methanol sequentially, 3% of formic acid in acetonitrile was used to elute arachidonic acid. The collected fraction was injected onto a reversed phase column at 30°C with mobile phase of acetonitrile/water (70 : 30, v/v) and detected by LC-MS/MS coupled with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The calibration curve ranged from 10 to 2500 ng/mL with sufficient linearity (r2 = 0.9999). The recoveries were in the range of 99.38% to 103.21% with RSD less than 6%. The limit of detection is 3 ng/mL. PMID:25873969

  16. 10. DETAIL OF BUILDER'S PLATE AT NORTH PORTAL. PLATE READS: ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    10. DETAIL OF BUILDER'S PLATE AT NORTH PORTAL. PLATE READS: 1889, BUILT BY THE BERLIN IRON BRIDGE CO. EAST BERLIN CONN. DOUGLAS & JARVIS PAT. APT. 16, 1878, AP'L 17, 1885. A.P. FORESMAN, WM. S. STARR, T.J. STREBEIGH, COMMISSIONERS. - Pine Creek Bridge, River Road spanning Pine Creek, Jersey Shore, Lycoming County, PA

  17. Measuring Ca²⁺ changes in multiwell format using the Fluorometric Imaging Plate Reader (FLIPR(®)).

    PubMed

    Marshall, Ian C B; Owen, Davina E; McNulty, Shaun

    2013-01-01

    The Fluorometric Imaging Plate Reader (FLIPR) has made a significant contribution to drug discovery programs. The key advantage of FLIPR over conventional plate readers is the ability to measure fluorescence emission from multiple wells (96 wells or 384 wells) simultaneously and with high temporal resolution. Consequently, FLIPR has been used extensively to record dynamic intracellular processes such as changes in intracellular Ca(2+) ion concentration, membrane potential, and pH. Since FLIPR is used to measure a functional response in cells, it is rapidly able to distinguish full agonists, partial agonists, and antagonists at a target of interest, making the system a valuable screening tool for interrogation of compound libraries. Automated FLIPR systems for ultra high throughput have also become available that employ integrated plate stackers, washers and specialized stages to allow users to shuttle cell and compound plates from incubators or storage magazines onto the FLIPR system itself. In this chapter generic methods for assessing intracellular Ca(2+) on the FLIPR are described. PMID:23007581

  18. Plate tectonics conserves angular momentum

    NASA Astrophysics Data System (ADS)

    Bowin, C.

    2010-03-01

    A new combined understanding of plate tectonics, Earth internal structure, and the role of impulse in deformation of the Earth's crust is presented. Plate accelerations and decelerations have been revealed by iterative filtering of the quaternion history for the Euler poles that define absolute plate motion history for the past 68 million years, and provide an unprecedented precision for plate angular rotation variations with time at 2-million year intervals. Stage poles represent the angular rotation of a plate's motion between adjacent Euler poles, and from which the maximum velocity vector for a plate can be determined. The consistent maximum velocity variations, in turn, yield consistent estimates of plate accelerations and decelerations. The fact that the Pacific plate was shown to accelerate and decelerate, implied that conservation of plate tectonic angular momentum must be globally conserved, and that is confirmed by the results shown here (total angular momentum ~1.4+27 kg m2 s-1). Accordingly, if a plate decelerates, other plates must increase their angular momentums to compensate. In addition, the azimuth of the maximum velocity vectors yields clues as to why the "bend" in the Emperor-Hawaiian seamount trend occurred near 46 Myr. This report summarizes processing results for 12 of the 14 major tectonic plates of the Earth (except for the Juan de Fuca and Philippine plates). Plate accelerations support the contention that plate tectonics is a product of torques that most likely are sustained by the sinking of positive density anomalies revealed by geoid anomalies of the degree 4-10 packet of the Earth's spherical harmonic coefficients. These linear positive geoid anomalies underlie plate subduction zones and are presumed due to phase changes in subducted gabbroic lithosphere at depth in the upper lower mantle (above 1200 km depth). The tectonic plates are pulled along by the sinking of these positive mass anomalies, rather than moving at near constant

  19. Plate tectonics, damage and inheritance.

    PubMed

    Bercovici, David; Ricard, Yanick

    2014-04-24

    The initiation of plate tectonics on Earth is a critical event in our planet's history. The time lag between the first proto-subduction (about 4 billion years ago) and global tectonics (approximately 3 billion years ago) suggests that plates and plate boundaries became widespread over a period of 1 billion years. The reason for this time lag is unknown but fundamental to understanding the origin of plate tectonics. Here we suggest that when sufficient lithospheric damage (which promotes shear localization and long-lived weak zones) combines with transient mantle flow and migrating proto-subduction, it leads to the accumulation of weak plate boundaries and eventually to fully formed tectonic plates driven by subduction alone. We simulate this process using a grain evolution and damage mechanism with a composite rheology (which is compatible with field and laboratory observations of polycrystalline rocks), coupled to an idealized model of pressure-driven lithospheric flow in which a low-pressure zone is equivalent to the suction of convective downwellings. In the simplest case, for Earth-like conditions, a few successive rotations of the driving pressure field yield relic damaged weak zones that are inherited by the lithospheric flow to form a nearly perfect plate, with passive spreading and strike-slip margins that persist and localize further, even though flow is driven only by subduction. But for hotter surface conditions, such as those on Venus, accumulation and inheritance of damage is negligible; hence only subduction zones survive and plate tectonics does not spread, which corresponds to observations. After plates have developed, continued changes in driving forces, combined with inherited damage and weak zones, promote increased tectonic complexity, such as oblique subduction, strike-slip boundaries that are subparallel to plate motion, and spalling of minor plates. PMID:24717430

  20. The antipsoriatic dimethyl-fumarate suppresses interferon-gamma -induced ICAM-1 and HLA-DR expression on hyperproliferative keratinocytes. Quantification by a culture plate-directed APAAP-ELISA technique.

    PubMed

    Sebok, B; Bonnekoh, B; Vetter, R; Schneider, I; Gollnick, H; Mahrle, G

    1998-01-01

    The derivatives of fumaric acid show antipsoriatic effects but details of the mechanism of action are largely unknown. The study focused on the effect of fumaric acid, dimethyl-fumarate, Zn-, Ca- and Mg-monoethyl-fumarate on the interferon-gamma (IFN-gamma)-induced expression of ICAM-1 and HLA-DR molecules on keratinocytes. Human hyperproliferative keratinocytes of the HaCaT cell line were exposed to IFN-gamma (10 U/ml) alone or in combination with fumaric acid and its derivatives for 48 hrs. The effect of fumarates was investigated semiquantitatively using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) method. Subsequently, the effect of dimethyl-fumarate, the main component of "fumaric acid therapy", was evaluated quantitatively by means of an APAAP-ELISA technique. The semiquantitative evaluation revealed that in the micromolar dose range investigated only dimethyl-fumarate demonstrated substantial growth inhibition and down-regulation of the cell surface markers. In the quantitative evaluation, dimethyl-fumarate significantly (pcultures. Thus, there is experimental evidence that dimethyl-fumarate may exert its antipsoriatic effect not only as an antiproliferative agent but also by down-regulation of ICAM-1 and HLA-DR molecules on hyperproliferative keratinocytes. PMID:9649687

  1. Uterine culture in mares.

    PubMed

    Brook, D

    1984-05-01

    A guarded, sterile swab is used to obtain samples for uterine culture. With the mare in stocks, the tail bandage and the perineum washed, the culture rod is introduced into the vagina with a gloved hand. After the rod is guided through the cervix, the guard cap is dislodged and the swab is rubbed along the endometrium, after which the rod is extracted. Samples for uterine culture should only be obtained during full estrus. Swabs should be directly plated onto agar within 2 hours of collection. Blood agar is appropriate for initial screening, but use of specialized types of agar expedites identification of microbes. Plates are incubated at 37 C and inspected for growth every 12 hours. The type and number of bacterial colonies should be coupled with the history and clinical signs in deciding on the necessity and type of treatment. Pure, heavy bacterial growth is usually accompanied by clinical signs of infection. Interpretation of the significance of moderate bacterial growth may be aided by cytologic examination of endometrial smears, made by rolling the swab onto a glass slide and staining with Diff - Quik . Large numbers of neutrophils indicate the need for antibiotic therapy. Mixed bacterial growth and variable numbers of neutrophils usually indicate faulty sampling technic. Microaerophilic or anaerobic cultures may aid diagnosis in cases of equivocal aerobic culture results. PMID:6377040

  2. Stepped conical zone plate antenna

    NASA Astrophysics Data System (ADS)

    Wiltse, James C.

    2001-07-01

    The Fresnel zone plate lens was invented and developed for optical frequencies. However, fabrication difficulties at the short optical wavelengths have prevented obtain good efficiencies. At longer microwave or millimeter-wavelengths fabrication is easier and phase correcting zone plate antennas have been used to obtain good efficiencies. This paper describes a new type of phase correcting zone plate having even better efficiency, namely a diffraction efficiency of 99 percent compared to a true lens, and an overall efficiency much better than a true lens. For the usual zone plate antenna employed at microwave or millimeter wavelengths, path length adjustment is accomplished by cutting different depths in a dielectric plate or by using two or more dielectrics having different dielectric constants. The new design uses a tilted cut in a dielectric plate, which more accurately matches the shape of a true lens and produces much lower phase error. The construction is still near and can be made for example, by a milling machine with a tilted bit. For a circular zone plate, the lens is a stepped conical or tapered shape. Because the phase steps are small, the far-field antenna pattern is excellent and sidelobe-levels are very low. Analysis of typical configurations will be given, showing that phase errors are small, lower than those for an eighth-wave corrected phase zone plate.

  3. ChooseMyPlate.gov

    MedlinePlus

    ... Day You don’t have to be an Olympic athlete to be active. Taking the stairs instead of the elevator, mowing the lawn, and gardening count too! TWITTER FEED 2 days ChooseMyPlate.gov @MyPlate Make family dinner more fun with a game or activity. Challenge kids to solve a riddle ...

  4. Micro-channel plate detector

    DOEpatents

    Elam, Jeffrey W.; Lee, Seon W.; Wang, Hsien -Hau; Pellin, Michael J.; Byrum, Karen; Frisch, Henry J.

    2015-09-22

    A method and system for providing a micro-channel plate detector. An anodized aluminum oxide membrane is provided and includes a plurality of nanopores which have an Al coating and a thin layer of an emissive oxide material responsive to incident radiation, thereby providing a plurality of radiation sensitive channels for the micro-channel plate detector.

  5. Present-day plate motions

    NASA Technical Reports Server (NTRS)

    Minster, J. B.; Jordan, T. H.

    1977-01-01

    A data set comprising 110 spreading rates, 78 transform fault azimuths and 142 earthquake slip vectors was inverted to yield a new instantaneous plate motion model, designated RM2. The mean averaging interval for the relative motion data was reduced to less than 3 My. A detailed comparison of RM2 with angular velocity vectors which best fit the data along individual plate boundaries indicates that RM2 performs close to optimally in most regions, with several notable exceptions. On the other hand, a previous estimate (RM1) failed to satisfy an extensive set of new data collected in the South Atlantic Ocean. It is shown that RM1 incorrectly predicts the plate kinematics in the South Atlantic because the presently available data are inconsistent with the plate geometry assumed in deriving RM1. It is demonstrated that this inconsistency can be remedied by postulating the existence of internal deformation with the Indian plate, although alternate explanations are possible.

  6. Impact damage of composite plates

    NASA Technical Reports Server (NTRS)

    Lal, K. M.; Goglia, G. L.

    1983-01-01

    A simple model to study low velocity transverse impact of thin plates made of fiber-reinforced composite material, in particular T300/5208 graphite-epoxy was discussed. This model predicts the coefficient of restitution, which is a measure of the energy absorbed by the target during an impact event. The model is constructed on the assumption that the plate is inextensible in the fiber direction and that the material is incompressible in the z-direction. Such a plate essentially deforms by shear, hence this model neglects bending deformations of the plate. The coefficient of restitution is predicted to increase with large interlaminar shear strength and low transverse shear modulus of the laminate. Predictions are compared with the test results of impacted circular and rectangular clamped plates. Experimentally measured values of the coefficient of restitution are found to agree with the predicted values within a reasonable error.

  7. SAMI Automated Plug Plate Configuration

    NASA Astrophysics Data System (ADS)

    Lorente, N. P. F.; Farrell, T.; Goodwin, M.

    2013-10-01

    The Sydney-AAO Multi-object Integral field spectrograph (SAMI) is a prototype wide-field system at the Anglo-Australian Telescope (AAT) which uses a plug-plate to mount its 13×61-core imaging fibre bundles (hexabundles) in the optical path at the telescope's prime focus. In this paper we describe the process of determining the positions of the plug-plate holes, where plates contain three or more stacked observation configurations. The process, which up until now has involved several separate processes and has required significant manual configuration and checking, is now being automated to increase efficiency and reduce error. This is carried out by means of a thin Java controller layer which drives the configuration cycle. This layer controls the user interface and the C++ algorithm layer where the plate configuration and optimisation is carried out. Additionally, through the Aladin display package, it provides visualisation and facilitates user verification of the resulting plates.

  8. Statistical tests of additional plate boundaries from plate motion inversions

    NASA Technical Reports Server (NTRS)

    Stein, S.; Gordon, R. G.

    1984-01-01

    The application of the F-ratio test, a standard statistical technique, to the results of relative plate motion inversions has been investigated. The method tests whether the improvement in fit of the model to the data resulting from the addition of another plate to the model is greater than that expected purely by chance. This approach appears to be useful in determining whether additional plate boundaries are justified. Previous results have been confirmed favoring separate North American and South American plates with a boundary located beween 30 N and the equator. Using Chase's global relative motion data, it is shown that in addition to separate West African and Somalian plates, separate West Indian and Australian plates, with a best-fitting boundary between 70 E and 90 E, can be resolved. These results are generally consistent with the observation that the Indian plate's internal deformation extends somewhat westward of the Ninetyeast Ridge. The relative motion pole is similar to Minster and Jordan's and predicts the NW-SE compression observed in earthquake mechanisms near the Ninetyeast Ridge.

  9. 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.

  10. Highly curved microchannel plates

    NASA Technical Reports Server (NTRS)

    Siegmund, O. H. W.; Cully, S.; Warren, J.; Gaines, G. A.; Priedhorsky, W.; Bloch, J.

    1990-01-01

    Several spherically curved microchannel plate (MCP) stack configurations were studied as part of an ongoing astrophysical detector development program, and as part of the development of the ALEXIS satellite payload. MCP pairs with surface radii of curvature as small as 7 cm, and diameters up to 46 mm have been evaluated. The experiments show that the gain (greater than 1.5 x 10 exp 7) and background characteristics (about 0.5 events/sq cm per sec) of highly curved MCP stacks are in general equivalent to the performance achieved with flat MCP stacks of similar configuration. However, gain variations across the curved MCP's due to variations in the channel length to diameter ratio are observed. The overall pulse height distribution of a highly curved surface MCP stack (greater than 50 percent FWHM) is thus broader than its flat counterpart (less than 30 percent). Preconditioning of curved MCP stacks gives comparable results to flat MCP stacks, but it also decreases the overall gain variations. Flat fields of curved MCP stacks have the same general characteristics as flat MCP stacks.

  11. Balanced Orifice Plate

    NASA Technical Reports Server (NTRS)

    Kelley, Anthony R. (Inventor); Buskirk, Paul D. (Inventor)

    2006-01-01

    An orifice plate for use in a conduit through which fluid flows is defined by a central circular region having a radius R, and a ring-shaped region surrounding the central circular region. The ring-shaped region has holes formed therethrough with those holes centered at each radius R thereof satisfying a relationship A(sub R)=al(X(sub R)V(sub R)(sup b)) where A(sub R) is a sum of areas of those holes having centers at radius R, X(sub R) is a flow coefficient at radius R, V(sub R) is a velocity of the fluid that is to flow through the conduit at radius R, b is a constant selected to make at least one process variable (associated with the fluid that is to flow through the conduit) approximately equal at each radius R, and a is a constant that is equal to (X(sub R)A(sub R)V(sub R)(sup b)) at each radius R.

  12. Mantle convection with plates and mobile, faulted plate margins.

    PubMed

    Zhong, S; Gurnis, M

    1995-02-10

    A finite-element formulation of faults has been incorporated into time-dependent models of mantle convection with realistic rheology, continents, and phase changes. Realistic tectonic plates naturally form with self-consistent coupling between plate and mantle dynamics. After the initiation of subduction, trenches rapidly roll back with subducted slabs temporarily laid out along the base of the transition zone. After the slabs have penetrated into the lower mantle, the velocity of trench migration decreases markedly. The inhibition of slab penetration into the lower mantle by the 670-kilometer phase change is greatly reduced in these models as compared to models without tectonic plates. PMID:17813909

  13. The Golosyiv plate archive digitisation

    NASA Astrophysics Data System (ADS)

    Sergeeva, T. P.; Sergeev, A. V.; Pakuliak, L. K.; Yatsenko, A. I.

    2007-08-01

    The plate archive of the Main Astronomical Observatory of the National Academy of Sciences of Ukraine (Golosyiv, Kyiv) includes about 85 000 plates which have been taken in various observational projects during 1950-2005. Among them are about 25 000 of direct northern sky area plates and more than 600 000 plates containing stellar, planetary and active solar formations spectra. Direct plates have a limiting magnitude of 14.0-16.0 mag. Since 2002 we have been organising the storage, safeguarding, cataloguing and digitization of the plate archive. The very initial task was to create the automated system for detection of astronomical objects and phenomena, search of optical counterparts in the directions of gamma-ray bursts, research of long period, flare and other variable stars, search and rediscovery of asteroids, comets and other Solar System bodies to improve the elements of their orbits, informational support of CCD observations and space projects, etc. To provide higher efficiency of this work we have prepared computer readable catalogues and database for 250 000 direct wide field plates. Now the catalogues have been adapted to Wide Field Plate Database (WFPDB) format and integrated into this world database. The next step will be adaptation of our catalogues, database and images to standards of the IVOA. Some magnitude and positional accuracy estimations for Golosyiv archive plates have been done. The photometric characteristics of the images of NGC 6913 cluster stars on two plates of the Golosyiv's double wide angle astrograph have been determined. Very good conformity of the photometric characteristics obtained with external accuracies of 0.13 and 0.15 mag. has been found. The investigation of positional accuracy have been made with A3± format fixed bed scanner (Microtek ScanMaker 9800XL TMA). It shows that the scanner has non-detectable systematic errors on the X-axis, and errors of ± 15 μm on the Y-axis. The final positional errors are about ± 2 μm (

  14. Turbidimetric assay for tetracyclines in feeds using a microtiter plate system.

    PubMed

    Brady, M S; Katz, S E

    1991-01-01

    The microtiter plate system for turbidimetric assay of chlortetracycline (CTC) and oxytetracycline (OTC) levels in feeds uses a 96 well microtiter plate, a multichannel pipette, and an ELISA reader to measure turbidity. Feeds are extracted for both tetracyclines using AOAC extraction systems. For CTC, the range of the standard curve is 0.001-0.005 microgram CTC/mL; for OTC, the range is 0.004-0.016 microgram OTC/mL. Repeatability of CTC assays, as shown by the coefficient of variation (CV), ranged from 0.54 to 5.65% for same-day assays and from 2.01 to 9.39% for assays on different days. For OTC, CVs ranged from 2.69 to 10.01% for same-day assays and 3.24 to 9.08% for different-day assays. Average recoveries for CTC were 108.7% for same-day assays and 106.8% for different-day assays; for OTC, average recoveries were 112.4% and 106.5% for same-day and different-day assays, respectively. PMID:1874689

  15. Difficulties encountered removing locked plates

    PubMed Central

    Raja, S; Imbuldeniya, AM; S, Garg; Groom, G

    2012-01-01

    INTRODUCTION Locked plates are commonly used to obtain fixation in periarticular and comminuted fractures. Their use has also gained popularity in repairing fractures in osteoporotic bone. These plates provide stable fixation and promote biological healing. Over the last 3 years, we have used over 150 locked plates with varying success to fix periarticular fractures involving mainly the knee and ankle. In this study, we report our clinical experience and the difficulties encountered when removing locked plates in adult patients with a variety of indications including implant failure, infection, non-union and a palpable symptomatic implant. METHODS A retrospective analysis was performed of patients enrolled prospectively into a database. Included in the study were 36 consecutive adult patients who each underwent the procedure of locked plate removal in a single inner city level 1 trauma centre. Data collected included primary indication for fixation, indication for implant removal, time of the implant in situ, grade of operating surgeon and difficulties encountered during the procedure. RESULTS Implant removal was associated with a complication rate of 47%. The major problems encountered were difficulty in removing the locked screws and the implant itself. A total of ten cold welded screws were found in eight cases. Removal was facilitated by high speed metal cutting burrs and screw removal sets in all but one case, where a decision was made to leave the plate in situ. CONCLUSIONS The majority of studies investigating implant removal and problems encountered in doing so report a relatively high complication rate. With the advent of locking plates and their growing popularity, difficulties are now being seen intra-operatively when removing them. There is a paucity of data, however, specifically directed at locking plate removal. We recommend that surgeons should be aware of the potential complications while removing locked plates. Fluoroscopic control and all

  16. High-throughput micro plate assays for screening flavonoid content and DPPH-scavenging activity in sorghum bran and flour

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rapid, 96-well microtiter assays were compared to conventional assays for quantifying total phenolic content, flavonoid content, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) in sorghum grain. The 96-well assays exhibited a correlation of >0.9 to the conventional assays. The 96-well assays allowed for ...

  17. High loading uranium fuel plate

    DOEpatents

    Wiencek, Thomas C.; Domagala, Robert F.; Thresh, Henry R.

    1990-01-01

    Two embodiments of a high uranium fuel plate are disclosed which contain a meat comprising structured uranium compound confined between a pair of diffusion bonded ductile metal cladding plates uniformly covering the meat, the meat having a uniform high fuel loading comprising a content of uranium compound greater than about 45 Vol. % at a porosity not greater than about 10 Vol. %. In a first embodiment, the meat is a plurality of parallel wires of uranium compound. In a second embodiment, the meat is a dispersion compact containing uranium compound. The fuel plates are fabricated by a hot isostatic pressing process.

  18. Plating on stainless steel alloys

    SciTech Connect

    Dini, J.W.; Johnson, H.R.

    1981-09-11

    Quantitative adhesion data are presented for a variety of electroplated stainless steel type alloys. Results show that excellent adhesion can be obtained by using a Wood's nickel strike or a sulfamate nickel strike prior to final plating. Specimens plated after Wood's nickel striking failed in the deposit rather than at the interface between the substrate and the coating. Flyer plate quantitative tests showed that use of anodic treatment in sulfuric acid prior to Wood's nickel striking even further improved adhesion. In contrast activation of stainless steels by immersion or cathodic treatment in hydrochloric acid resulted in very reduced bond strengths with failure always occurring at the interface between the coating and substrate.

  19. The multigap resistive plate chamber

    SciTech Connect

    Zeballos, E. Cerron; Crotty, I.; Hatzifotiadou, D.; Valverde, J. Lamas; Neupane, S.; Williams, M. C. S.; Zichichi, A.

    2015-02-03

    The paper describes the multigap resistive plate chamber (RPC). This is a variant of the wide gap RPC. However it has much improved time resolution, while keeping all the other advantages of the wide gap RPC design.

  20. Quaternions as astrometric plate constants

    NASA Technical Reports Server (NTRS)

    Jefferys, William H.

    1987-01-01

    A new method for solving problems in relative astrometry is proposed. In it, the relationship between the measured quantities and the components of the position vector of a star is modeled using quaternions, in effect replacing the plate constants of a standard four-plate-constant solution with the four components of a quaternion. The method allows a direct solution for the position vectors of the stars, and hence for the equatorial coordinates. Distortions, magnitude, and color effects are readily incorporated into the formalism, and the method is directly applicable to overlapping-plate problems. The advantages of the method include the simplicity of the resulting equations, their freedom from singularities, and the fact that trigonometric functions and tangential point transformations are not needed to model the plate material. A global solution over the entire sky is possible.

  1. Carbon-assisted flyer plates

    DOEpatents

    Stahl, D.B.; Paisley, D.L.

    1994-04-12

    A laser driven flyer plate is described utilizing an optical fiber connected to a laser. The end of the optical fiber has a layer of carbon and a metal layer deposited onto it. The carbon layer provides the laser induced plasma which is superior to the plasma produced from most metals. The carbon layer plasma is capable of providing a flatter flyer plate, converting more of the laser energy to driving plasma, promoting a higher flyer plate acceleration, and providing a more uniform pulse behind the plate. In another embodiment, the laser is in optical communication with a substrate onto which a layer of carbon and a layer of metal have been deposited. 2 figures.

  2. Soft x-ray microscope with zone plates at UVSOR

    NASA Astrophysics Data System (ADS)

    Watanabe, Norio; Shimanuki, Yoshio; Taniguchi, Mieko; Kihara, Hiroshi

    1993-01-01

    A soft x-ray microscope with zone plates was set up at UVSOR (Okazaki, Japan). A 0.41 micrometers line and space pattern was clearly distinguished using an objective zone plate with the outermost zone width of 0.41 micrometers . Modulation transfer functions were measured at wavelengths of 3.1 nm and 5.4 nm, and compared with theoretical calculations. Considering the resolution of a microchannel plate used as a detector, the agreement is fairly good. With this microscope, some biological specimens such as diatoms, spicule of trepang, crab and rabbit muscles, human blood cells, human chromosomes, and magnetotactic bacterium were observed at 3.1 nm and 5.4 nm. With an environmental chamber (wet cell) using polypropylene foils as windows, wet specimens were observed at a wavelength of 4.6 nm. Images of spicule of trepang, human blood cell, and cultured protoplast of plant cell stained by methyl mercury were observed with good contrast.

  3. Pulse plating of nickel deposits

    SciTech Connect

    Stimetz, C.J.; Stevenson, M.F.

    1980-02-01

    Pulse plated and conventional nickel deposits have been compared for differences in morphology, mechanical properties, and microstructure. The deposits were obtained from nickel sulfamate, nickel chloride, and Watts nickel plating solutions. No significant differences were found in the direct and pulse current deposits from the sulfamate and chloride solutions; however, significant differences in microstructure, yield strength, and microhardness were observed in deposits from the Watts nickel solution.

  4. Microtissue Culture Plaque Assay for Herpesvirus saimiri

    PubMed Central

    Chan, Emerson W.; Dunkel, Virginia C.

    1973-01-01

    A microtissue culture method for the plaque assay of Herpesvirus saimiri has been developed. Virus titrations carried out in Microtest II tissue culture plates (Falcon) yielded reproducible results that agreed well with those obtained by employing macrocultures. The described method is quantitative, reproducible, economical, and suitable for routine assay of large numbers of virus samples. Images PMID:4201642

  5. Impact on multilayered composite plates

    NASA Technical Reports Server (NTRS)

    Kim, B. S.; Moon, F. C.

    1977-01-01

    Stress wave propagation in a multilayer composite plate due to impact was examined by means of the anisotropic elasticity theory. The plate was modelled as a number of identical anisotropic layers and the approximate plate theory of Mindlin was then applied to each layer to obtain a set of difference-differential equations of motion. Dispersion relations for harmonic waves and correction factors were found. The governing equations were reduced to difference equations via integral transforms. With given impact boundary conditions these equations were solved for an arbitrary number of layers in the plate and the transient propagation of waves was calculated by means of a Fast Fourier Transform algorithm. The multilayered plate problem was extended to examine the effect of damping layers present between two elastic layers. A reduction of the interlaminar normal stress was significant when the thickness of damping layer was increased but the effect was mostly due to the softness of the damping layer. Finally, the problem of a composite plate with a crack on the interlaminar boundary was formulated.

  6. How mantle slabs drive plate tectonics.

    PubMed

    Conrad, Clinton P; Lithgow-Bertelloni, Carolina

    2002-10-01

    The gravitational pull of subducted slabs is thought to drive the motions of Earth's tectonic plates, but the coupling between slabs and plates is not well established. If a slab is mechanically attached to a subducting plate, it can exert a direct pull on the plate. Alternatively, a detached slab may drive a plate by exciting flow in the mantle that exerts a shear traction on the base of the plate. From the geologic history of subduction, we estimated the relative importance of "pull" versus "suction" for the present-day plates. Observed plate motions are best predicted if slabs in the upper mantle are attached to plates and generate slab pull forces that account for about half of the total driving force on plates. Slabs in the lower mantle are supported by viscous mantle forces and drive plates through slab suction. PMID:12364804

  7. Measurement of TACE Activity in Extracts from Cultured Cells

    PubMed Central

    Xu, Xiuling; Padilla, Mabel T.; Lin, Yong

    2016-01-01

    In cigarette smoke–induced and inflammation-associated lung cancer development, cigarette smoke extract (CSE) activates tumor necrosis factor-alpha (TNF-α) secretion from macrophages. TNF-α converting enzyme (TACE), also known as α-Secretase or ADAM17 (A Disintegrin and Metalloprotease), is a member of the ADAM family of metalloproteases. TACE mediated ectodomain shedding leads to the conversion of the inactive TNF-α precursor into the active mature pro-inflammatory cytokine. The SensoLyte 520 TACE (α-Secretase) Activity Assay Kit was used to detect TACE activity in CSE-activated macrophages. This assay is reliable, reproducible and easy to carry out in 96 well plate format.

  8. Plate mode velocities in graphite/epoxy plates

    NASA Technical Reports Server (NTRS)

    Prosser, W. H.; Gorman, M. R.

    1994-01-01

    Measurements of the velocities of the extensional and flexural plate modes were made along three directions of propagation in four graphite/epoxy composite plates. The acoustic signals were generated by simulated acoustic emission events (pencil lead breaks or Hsu-Neilson sources) and detected by by broadband ultrasonic transducers. The first arrival of the extensional plate mode, which is nondispersive at low frequencies, was measured at a number of different distances from the source along the propagation direction of interest. The velocity was determined by plotting the distance versus arrival time and computing its slope. Because of the large dispersion of the flexural mode, a Fourier phase velocity technique was used to characterize this mode. The velocity was measured up to a frequency of 160 kHz. Theoretical predictions of the velocities of these modes were also made and compared with experimental observations. Classical plate theory yields good agreement with the measured extensional velocities. For predictions of the dispersion of the flexural mode, Mindlin plates theory, which includes the effects of shear deformation and rotatory inertia was shown to give better agreement with the experimental measurements.

  9. Simplified 48-hour IMVic test: an agar plate method.

    PubMed

    Powers, E M; Latt, T G

    1977-09-01

    An agar plate method was developed for the performance of the IMVic (indole, methyl red, Voges-Proskauer, and citrate) tests in lieu of the conventional tubed liquid media. By modifying the composition of the media and adding agar, a single "X"-compartmented petri dish was prepared containing all four IMVic test media. Ease of performance and simplification of the test were achieved by inoculating all four media simultaneously from a single colony (single inoculum) on eosin-methylene blue agar. Tests with 87 cultures, representing 7 genera in the family Enterobacteriaceae, were completed with typical (correct) IMVic patterns for all cultures within 48 h. Parallel tests with conventional media showed that the agar plate method was superior, more sensitive, faster, and simpler to perform, and less time was required to identify Escherichia coli by 72 h. PMID:334074

  10. Dehydration improves cryopreservation of mat rush (Juncus decipiens Nakai) basal stem buds on cryo-plates.

    PubMed

    Niino, T; Yamamoto, S I; Fukui, K; Castillo Martinez, C R; Arizaga, M V; Matsumoto, T; Engelmann, F

    2013-01-01

    Two cryopreservation procedures using aluminium cryo-plates, termed V-Cryo-plate and D-Cryo-plate, were successfully developed for in vitro mat rush (Juncus decipiens Nakai) basal stem buds. Multiple stems induced in liquid MS medium containing 8.9 μM BA by roller culture were cut into small clumps, plated on solid MS medium and cultured for 1 week at 25 degree C. Clumps that had produced many buds were cold-hardened at 5 degree C for 1-2 months. The buds with basal stems were dissected from small clumps and precultured overnight at 25 degree C on solid MS medium containing 0.3 M sucrose. Precultured buds were placed on aluminium cryo-plates and embedded in calcium alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25 degree C in loading solution (2 M glycerol + 1.0 M sucrose). In the D-Cryo-plate procedure, the buds were dehydrated to 27-25% moisture content (fresh weight) by placing the cryo-plates in the air current of a laminar flow cabinet for 2 to 3 h. In the V-Cryo-plate procedure, buds were dehydrated by immersing the cryo-plates in PVS2 vitrification solution for 40 min at 25 degree C. In both procedures, cooling was performed by placing the cryo-plates in uncapped cryotubes, which were immersed in liquid nitrogen. For rewarming, cryo-plates were immersed in medium with 1.0 M sucrose for 20 min at room temperature. Regrowth of cryopreserved buds of line 'Kitakei 2' using D-Cryo-plate and V-Cryo-plate procedures, was 90% and 80%, respectively. The two procedures were applied to 20 additional mat rush lines. Using the V-Cryo-plate procedure resulted in regrowth ranging between 13.3 and 86.7%, with an average of 52.5%. The D-Cryo-plate led to regrowth ranging between 73.3 and 96.7%, with an average of 86.3%. The D-Cryo-plate procedure will facilitate cryostorage of mat rush germplasm. PMID:24441366

  11. The tectonic plates are shifting: cultural change vs. mural dyslexia.

    PubMed

    Cohn, Kenneth; Friedman, Leonard H; Allyn, Thomas R

    2007-01-01

    In response to a rapidly changing healthcare marketplace, a variety of new business models have arisen, including new specialties (hospitalists), selective care (concierge medicine), and joint ventures (ambulatory surgical centers, specialty hospitals), some with hospitals and others with independent vendors. Since both hospitals and physicians are feeling the squeeze of rising expenses, burdensome regulations, heightened consumer expectations, and stagnant or decreasing reimbursement, the response to global economic competition and the need to improve clinical and financial outcomes can bring physicians and hospitals together rather than drive them farther apart. In response to perceived threats, physicians and hospital executives can engage in defensive reasoning that may feel protective but can also lead to mural dyslexia, the inability or unwillingness to see the handwriting on the wall. The strategies of positive deviance (finding solutions that already exist in the community rather than importing best practices), appreciative inquiry (building on success rather than relying solely on root-cause analyses of problems), and structured dialogue (allowing practicing physicians to articulate clinical priorities rather than assuming they lack the maturity and will to come to consensus) are field-tested approaches that allow hospital leaders to engage practicing physicians and that can help both parties work more interdependently to improve patient care in a dynamically changing environment. Physician-hospital collaboration based on transparency, active listening, and prompt implementation can offer sustainable competitive advantage to those willing to embark on a lifetime learning journey. PMID:17907689

  12. Wide-Field Plate Database: Included Ukrainian Plate Catalogues

    NASA Astrophysics Data System (ADS)

    Tsvetkova, Katya; Sergeeva, Tetyana

    2007-08-01

    We present the basic information for the archives of the Main Astronomical Observatory of the National Academy of Sciences of Ukraine, included into the Wide-Field Plate Database last two years with total number of plates - 11260. The plates were obtained with the 0.40m Double Wide-angle Astrograph (DWA); 10cm, 11cm and 15cm Three Cameras Astrograph (TCA) and 12 cm Double Short focus Astrograph (DSA) in the period 1949-1998 in the frames of the following observing programmes: Photographic Survey of the Northern Sky; Investigation of the kinematics and the structure in the main meridian section of the Galaxy; selection of reference stars, minor planets observations, comet investigations.

  13. Pre-Plated Cell Lines for ADMETox Applications in the Pharmaceutical Industry.

    PubMed

    Torres, Francisco M; Sáfár, Zsolt; Vázquez-Sánchez, Maria A; Kurunczi, Anita; Kis, Emese; Magnan, Rémi; Jani, Márton; Nicolás, Josep Oriol; Krajcsi, Péter

    2015-01-01

    Membrane transporters significantly modulate membrane permeability of endobiotics and xenobiotics, such as bile acids and drugs, respectively. Various in vitro methods have been established for both ATP-binding cassette (ABC) transporters to examine cellular efflux and uptake, and for solute carriers (SLC) to examine cellular uptake of substrates. Cell-based systems are the models of choice to test drug-transporter interactions as well as drug-drug interactions for research and regulatory purposes, albeit, for low passive permeability substrates of ABC transporters, vesicular uptake assays are also recommended. Commercially available pre-plated cells (e.g., immortalized or transfected) offer a useful alternative to in-house cell culture. Three main methods are known to manufacture pre-plated cultures: regular culture medium with vacuum seal, cryopreserved delivery, and the solid shipping media technology. The regular culture medium and the solid shipping media technologies provide ready-to-use models for end users. Models expressing a broad selection of transporters are available in pre-plated formats for absorption, distribution, metabolism, excretion, and toxicity (ADMETox) studies. Conversely, the application and utility of pre-plated cultures coupled with personal experiences have not been extensively covered in published research papers or reviews, despite availability and significant use of pre-plated products in the pharmaceutical industry. In this overview, we will briefly describe: 1) in vitro tools commonly used for ADMETox testing; 2) methods employed in manufacturing, shipment and preparation of pre-plated cell lines; 3) cell-membrane barrier models currently available in pre-plated format to reproduce passage restriction of physiological barriers to certain compounds; and 4) recommended pre-plated cell lines overexpressing uptake transporters for ADMETox applications. PMID:26250397

  14. 30 CFR 20.13 - Approval plate.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... plate shall be satisfactory to MSHA. (a) Purpose of approval plate. The approval plate is a label which.... By it the manufacturer can point out that his lamp complies with specifications of MSHA and that it... place MSHA's approval plate on his lamp obligates the manufacturer to maintain the quality of...

  15. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 6 2013-10-01 2013-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the...

  16. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 6 2010-10-01 2010-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the...

  17. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 6 2014-10-01 2014-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the...

  18. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 6 2012-10-01 2012-10-01 false Plate specifications. 451.23 Section 451.23... SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The safety approval plate must be of the size and in the...

  19. APPLICATION OF A NEW MICROCULTURING TECHNIQUE TO ASSESS THE EFFECTS OF TEMPERATURE AND SALINITY ON SPECIFIC GROWTH RATES OF SIX SYMBIODINIUM ISOLATES

    EPA Science Inventory

    A simple micro-culturing technique is described for determining specific growth rates of cultured Symbiodinium spp. Micro-cultures were prepared by transferring 200 L fresh test medium containing 2–10 Symbiodinium cells to wells of a flat bottom 96-well plate. Cultures were incub...

  20. Beyond plate tectonics - Looking at plate deformation with space geodesy

    NASA Technical Reports Server (NTRS)

    Jordan, Thomas H.; Minster, J. Bernard

    1988-01-01

    The requirements that must be met by space-geodetic systems in order to constrain the horizontal secular motions associated with the geological deformation of the earth's surface are explored. It is suggested that in order to improve existing plate-motion models, the tangential components of relative velocities on interplate baselines must be resolved to an accuracy of less than 3 mm/yr. Results indicate that measuring the velocities between crustal blocks to + or - 5 mm/yr on 100-km to 1000-km scales can produce geologically significant constraints on the integrated deformation rates across continental plate-boundary zones such as the western United States.

  1. Plating on some difficult-to-plate metals and alloys

    SciTech Connect

    Dini, J.W.; Johnson, H.R.

    1980-02-01

    Electrodeposition of coatings on metals such as beryllium, beryllium-copper, Kovar, lead, magnesium, thorium, titanium, tungsten, uranium, zirconium, and their alloys can be problematic. This is due in most cases to a natural oxide surface film that readily reforms after being removed. The procedures we recommend for plating on these metals rely on replacing the oxide film with a displacement coating, or etching to allow mechanical keying between the substrate and plated deposit. The effectiveness of the procedures is demonstrated by interface bond strengths found in ring-shear and conical-head tensile tests.

  2. Urine culture

    MedlinePlus

    Culture and sensitivity - urine ... when urinating. You also may have a urine culture after you have been treated for an infection. ... when bacteria or yeast are found in the culture. This likely means that you have a urinary ...

  3. Stool Culture

    MedlinePlus

    ... Bacterial Culture, stool; Feces Culture Formal name: Enteric Pathogens Culture, stool Related tests: Ova and Parasite Exam , ... Shiga toxin-producing Escherichia coli , Widal Test , Gastrointestinal Pathogens Panel All content on Lab Tests Online has ...

  4. Fecal culture

    MedlinePlus

    Stool culture; Culture - stool ... stool tests are done in addition to the culture, such as: Gram stain of stool Fecal smear ... Giannella RA. Infectious enteritis and proctocolitis and bacterial food poisoning. In: Feldman M, Friedman LS, Brandt LJ, ...

  5. Plates for vacuum thermal fusion

    DOEpatents

    Davidson, James C.; Balch, Joseph W.

    2002-01-01

    A process for effectively bonding arbitrary size or shape substrates. The process incorporates vacuum pull down techniques to ensure uniform surface contact during the bonding process. The essence of the process for bonding substrates, such as glass, plastic, or alloys, etc., which have a moderate melting point with a gradual softening point curve, involves the application of an active vacuum source to evacuate interstices between the substrates while at the same time providing a positive force to hold the parts to be bonded in contact. This enables increasing the temperature of the bonding process to ensure that the softening point has been reached and small void areas are filled and come in contact with the opposing substrate. The process is most effective where at least one of the two plates or substrates contain channels or grooves that can be used to apply vacuum between the plates or substrates during the thermal bonding cycle. Also, it is beneficial to provide a vacuum groove or channel near the perimeter of the plates or substrates to ensure bonding of the perimeter of the plates or substrates and reduce the unbonded regions inside the interior region of the plates or substrates.

  6. Rapid throughput analysis demonstrates that chemicals with distinct seizurogenic mechanisms differentially alter Ca2+ dynamics in networks formed by hippocampal neurons in culture.

    PubMed

    Cao, Zhengyu; Zou, Xiaohan; Cui, Yanjun; Hulsizer, Susan; Lein, Pamela J; Wulff, Heike; Pessah, Isaac N

    2015-04-01

    Primary cultured hippocampal neurons (HN) form functional networks displaying synchronous Ca(2+) oscillations (SCOs) whose patterns influence plasticity. Whether chemicals with distinct seizurogenic mechanisms differentially alter SCO patterns was investigated using mouse HN loaded with the Ca(2+) indicator fluo-4-AM. Intracellular Ca(2+) dynamics were recorded from 96 wells simultaneously in real-time using fluorescent imaging plate reader. Although quiescent at 4 days in vitro (DIV), HN acquired distinctive SCO patterns as they matured to form extensive dendritic networks by 16 DIV. Challenge with kainate, a kainate receptor (KAR) agonist, 4-aminopyridine (4-AP), a K(+) channel blocker, or pilocarpine, a muscarinic acetylcholine receptor agonist, caused distinct changes in SCO dynamics. Kainate at <1 µM produced a rapid rise in baseline Ca(2+) (Phase I response) associated with high-frequency and low-amplitude SCOs (Phase II response), whereas SCOs were completely repressed with >1 µM kainate. KAR competitive antagonist CNQX [6-cyano-7-nitroquinoxaline-2,3-dione] (1-10 µM) normalized Ca(2+) dynamics to the prekainate pattern. Pilocarpine lacked Phase I activity but caused a sevenfold prolongation of Phase II SCOs without altering either their frequency or amplitude, an effect normalized by atropine (0.3-1 µM). 4-AP (1-30 µM) elicited a delayed Phase I response associated with persistent high-frequency, low-amplitude SCOs, and these disturbances were mitigated by pretreatment with the KCa activator SKA-31 [naphtho[1,2-d]thiazol-2-ylamine]. Consistent with its antiepileptic and neuroprotective activities, nonselective voltage-gated Na(+) and Ca(2+) channel blocker lamotrigine partially resolved kainate- and pilocarpine-induced Ca(2+) dysregulation. This rapid throughput approach can discriminate among distinct seizurogenic mechanisms that alter Ca(2+) dynamics in neuronal networks and may be useful in screening antiepileptic drug candidates. PMID:25583085

  7. Safeguards Culture

    SciTech Connect

    Frazar, Sarah L.; Mladineo, Stephen V.

    2012-07-01

    The concepts of nuclear safety and security culture are well established; however, a common understanding of safeguards culture is not internationally recognized. Supported by the National Nuclear Security Administration, the authors prepared this report, an analysis of the concept of safeguards culture, and gauged its value to the safeguards community. The authors explored distinctions between safeguards culture, safeguards compliance, and safeguards performance, and evaluated synergies and differences between safeguards culture and safety/security culture. The report concludes with suggested next steps.

  8. Insert metering plates for gas turbine nozzles

    DOEpatents

    Burdgick, Steven S.; Itzel, Gary; Chopra, Sanjay; Abuaf, Nesim; Correia, Victor H.

    2004-05-11

    The invention comprises a metering plate which is assembled to an impingement insert for use in the nozzle of a gas turbine. The metering plate can have one or more metering holes and is used to balance the cooling flow within the nozzle. A metering plate with multiple holes reduces static pressure variations which result from the cooling airflow through the metering plate. The metering plate can be assembled to the insert before or after the insert is inserted into the nozzle.

  9. Surface crack problems in plates

    NASA Technical Reports Server (NTRS)

    Joseph, P. F.; Erdogan, F.

    1989-01-01

    The mode I crack problem in plates under membrane loading and bending is reconsidered. The purpose is to examine certain analytical features of the problem further and to provide some new results. The formulation and the results given by the classical and the Reissner plate theories for through and part-through cracks are compared. For surface cracks the three-dimensional finite element solution is used as the basis of comparison. The solution is obtained and results are given for the crack/contact problem in a plate with a through crack under pure bending and for the crack interaction problem. Also, a procedure is developed to treat the problem of subcritical crack growth and to trace the evolution of the propagating crack.

  10. Dual polarization flat plate antenna

    NASA Astrophysics Data System (ADS)

    Kelly, Kenneth C.

    Rectangular waveguides with radiating slots are used in groups to form planar array microwave antennas with large apertures and small depth. Such flat plate antennas are widely used on spacecraft and aircraft. Typically, flat plate antennas provide fixed linear polarization. The present paper describes a new flat plate antenna which produces two coincident beams that are distinguished by their orthogonal linear polarizations. The antenna has two ports, one for each of the coicident beams. Completely external to the antenna, connecting a simple network to those terminal ports enables the antenna to provide right circular polarization from one port and left from the other. A different external network enables the antenna to have arbitrarily adjustable polarizations.

  11. Achromatic axially symmetric wave plate.

    PubMed

    Wakayama, Toshitaka; Komaki, Kazuki; Otani, Yukitoshi; Yoshizawa, Toru

    2012-12-31

    An achromatic axially symmetric wave plate (AAS-WP) is proposed that is based on Fresnel reflections. The wave plate does not introduce spatial dispersion. It provides retardation in the wavelength domain with an axially symmetric azimuthal angle. The optical configuration, a numerical simulation, and the optical properties of the AAS-WP are described. It is composed of PMMA. A pair of them is manufactured on a lathe. In the numerical simulation, the achromatic angle is estimated and is used to design the devices. They generate an axially symmetric polarized beam. The birefringence distribution is measured in order to evaluate the AAS-WPs. PMID:23388751

  12. Novel approach of high cell density recombinant bioprocess development: Optimisation and scale-up from microlitre to pilot scales while maintaining the fed-batch cultivation mode of E. coli cultures

    PubMed Central

    2010-01-01

    Background Bioprocess development of recombinant proteins is time consuming and laborious as many factors influence the accumulation of the product in the soluble and active form. Currently, in most cases the developmental line is characterised by a screening stage which is performed under batch conditions followed by the development of the fed-batch process. Performing the screening already under fed-batch conditions would limit the amount of work and guarantee that the selected favoured conditions also work in the production scale. Results Here, for the first time, high throughput multifactorial screening of a cloning library is combined with the fed-batch technique in 96-well plates, and a strategy is directly derived for scaling to bioreactor scale. At the example of a difficult to express protein, an RNase inhibitor, it is demonstrated that screening of various vector constructs and growth conditions can be performed in a coherent line by (i) applying a vector library with promoters and ribosome binding sites of different strength and various fusion partners together with (ii) an early stage use of the fed-batch technology. It is shown that the EnBase® technology provides an easy solution for controlled cultivation conditions in the microwell scale. Additionally the high cell densities obtained provide material for various analyses from the small culture volumes. Crucial factors for a high yield of the target protein in the actual case were (i) the fusion partner, (ii) the use of of a mineral salt medium together with the fed-batch technique, and (iii) the preinduction growth rate. Finally, it is shown that the favorable conditions selected in the microwell plate and shake flask scales also work in the bioreactor. Conclusions Cultivation media and culture conditions have a major impact on the success of a screening procedure. Therefore the application of controlled cultivation conditions is pivotal. The consequent use of fed-batch conditons from the first

  13. Mammosphere culture of cancer stem cells in a microfluidic device

    NASA Astrophysics Data System (ADS)

    Saadin, Katayoon; White, Ian M.

    2012-03-01

    It is known that tumor-initiating cells with stem-like properties will form spherical colonies - termed mammospheres - when cultured in serum-free media on low-attachment substrates. Currently this assay is performed in commercially available 96-well trays with low-attachment surfaces. Here we report a novel microsystem that features on-chip mammosphere culture on low attachment surfaces. We have cultured mammospheres in this microsystem from well-studied human breast cancer cell lines. To enable the long-term culture of these unattached cells, we have integrated diffusion-based delivery columns that provide zero-convection delivery of reagents, such as fresh media, staining agents, or drugs. The multi-layer system consists of parallel cell-culture chambers on top of a low-attachment surface, connected vertically with a microfluidic reagent delivery layer. This design incorporates a reagent reservoir, which is necessary to reduce evaporation from the cell culture micro-chambers. The development of this microsystem will lead to the integration of mammosphere culture with other microfluidic functions, including circulating tumor cell recovery and high throughput drug screening. This will enable the cancer research community to achieve a much greater understanding of these tumor initiating cancer stem cells.

  14. Cyclooxygenases and prostaglandin E2 receptors in growth plate chondrocytes in vitro and in situ--prostaglandin E2 dependent proliferation of growth plate chondrocytes.

    PubMed

    Brochhausen, Christoph; Neuland, Pia; Kirkpatrick, C James; Nüsing, Rolf M; Klaus, Günter

    2006-01-01

    Prostaglandin E2 (PGE2) plays an important role in bone development and metabolism. To interfere therapeutically in the PGE2 pathway, however, knowledge about the involved enzymes (cyclooxygenases) and receptors (PGE2 receptors) is essential. We therefore examined the production of PGE2 in cultured growth plate chondrocytes in vitro and the effects of exogenously added PGE2 on cell proliferation. Furthermore, we analysed the expression and spatial distribution of cyclooxygenase (COX)-1 and COX-2 and PGE2 receptor types EP1, EP2, EP3 and EP4 in the growth plate in situ and in vitro. PGE2 synthesis was determined by mass spectrometry, cell proliferation by DNA [3H]-thymidine incorporation, mRNA expression of cyclooxygenases and EP receptors by RT-PCR on cultured cells and in homogenized growth plates. To determine cellular expression, frozen sections of rat tibial growth plate and primary chondrocyte cultures were stained using immunohistochemistry with polyclonal antibodies directed towards COX-1, COX-2, EP1, EP2, EP3, and EP4. Cultured growth plate chondrocytes transiently secreted PGE2 into the culture medium. Although both enzymes were expressed in chondrocytes in vitro and in vivo, it appears that mainly COX-2 contributed to PGE2-dependent proliferation. Exogenously added PGE2 stimulated DNA synthesis in a dose-dependent fashion and gave a bell-shaped curve with a maximum at 10-8 M. The EP1/EP3 specific agonist sulprostone and the EP1-selective agonist ONO-D1-004 increased DNA synthesis. The effect of PGE2 was suppressed by ONO-8711. The expression of EP1, EP2, EP3, and EP4 receptors in situ and in vitro was observed; EP2 was homogenously expressed in all zones of the growth plate in situ, whereas EP1 expression was inhomogenous, with spared cells in the reserve zone. In cultured cells these four receptors were expressed in a subset of cells only. The most intense staining for the EP1 receptor was found in polygonal cells surrounded by matrix. Expression of

  15. Learning Culture.

    ERIC Educational Resources Information Center

    Jones, David

    Adult and continuing education in the arts can and does play a role in the development of cultural identity. Dimensions of culture include ethnicity, location, age, social class, and time. This definition of culture leads to the conclusion that cultures are generally small and are dynamic rather than static. Research shows that individuals in what…

  16. Culture matters.

    PubMed

    Arif, Zeba

    Zebaa Arif reflects on changes during her career as a mental health nurse in relation to cultural care issues: Cultural awareness is becoming embedded in patient care. All aspects of care are influenced by cultural beliefs and should form part of assessment. Leadership is essential in influencing cultural care, as is organisational commitment. PMID:16262169

  17. Overriding Plate Controls on Subduction Zone Evolution

    NASA Astrophysics Data System (ADS)

    Sharples, W. K.; Jadamec, M. A.; Moresi, L. N.; Capitanio, F. A.

    2014-12-01

    Seismic data, rock deformation experiments, and geochemical studies indicate variability in the thickness, buoyancy, and strength of the lithosphere at plate boundaries. However, geodynamic models of subduction commonly either omit an overriding plate or do not investigate role of the variation in overriding plate properties on the subduction evolution. We present time-dependent numerical models of subduction that vary the overriding plate thickness, strength, and density and allow for a plate interface that evolves with time via an anisotropic brittle failure rheology. We examine the emergence of (a) asymmetric versus symmetric subduction, (b) trench retreat versus advance, (c) subduction zone geometry, (d) slab stagnation versus penetration into the lower mantle, and (e) flat slab subduction. The majority of the models result in sustained asymmetric subduction. The models demonstrate that trench retreat is correlated with a thin overriding plate, whereas, trench advance is correlated with a thick and/or strong overriding plate. Slab dip, measured at a depth below the plate boundary interface, has a negative correlation with an increase in overriding plate thickness. Overriding plate thickness exerts a first order control over slab penetration into the lower mantle, with penetration most commonly occurring in models with a thick overriding plate. Periods of flat slab subduction occur with thick, strong overriding plates producing strong plate boundary interface coupling. The results provide insight into how the overriding plate plays a role in establishing advancing and retreating subduction, as well as providing an explanation for the variation of slab geometry observed in subduction zones on Earth.

  18. Elastic stability of thick auxetic plates

    NASA Astrophysics Data System (ADS)

    Lim, Teik-Cheng

    2014-04-01

    Auxetic materials and structures exhibit a negative Poisson’s ratio while thick plates encounter shear deformation, which is not accounted for in classical plate theory. This paper investigates the effect of a negative Poisson’s ratio on thick plates that are subjected to buckling loads, taking into consideration the shear deformation using Mindlin plate theory. Using a highly accurate shear correction factor that allows for the effect of Poisson’s ratio, the elastic stability of circular and square plates are evaluated in terms of dimensionless parameters, namely the Mindlin-to-Kirchhoff critical buckling load ratio and Mindlin critical buckling load factors. Results for thick square plates reveal that both parameters increase as the Poisson’s ratio becomes more negative. In the case of thick circular plates, the Mindlin-to-Kirchhoff critical buckling load ratios and the Mindlin critical buckling load factors increase and decrease, respectively, as the Poisson’s ratio becomes more negative. The results obtained herein show that thick auxetic plates behave as thin conventional plates, and therefore suggest that the classical plate theory can be used to evaluate the elastic stability of thick plates if the Poisson’s ratio of the plate material is sufficiently negative. The results also suggest that materials with highly negative Poisson’s ratios are recommended for square plates, but not circular plates, that are subjected to buckling loads.

  19. Petroleum occurrences and plate tectonics

    SciTech Connect

    Olenin, V.B.; Sokolov, B.A.

    1983-01-01

    This paper analyzes the mechanisms of petroleum formation and petroleum accumulation proposed in recent years by some Russian and foreign investigators from the viewpoint of the new global or plate tectonics. On the basis of discussion and the facts, the authors conclude that the mechanisms proposed are in contradiction to reality and their use in practical application is at least premature.

  20. Plated wire random access memories

    NASA Technical Reports Server (NTRS)

    Gouldin, L. D.

    1975-01-01

    A program was conducted to construct 4096-work by 18-bit random access, NDRO-plated wire memory units. The memory units were subjected to comprehensive functional and environmental tests at the end-item level to verify comformance with the specified requirements. A technical description of the unit is given, along with acceptance test data sheets.

  1. Corrosion resistant metallic bipolar plate

    DOEpatents

    Brady, Michael P.; Schneibel, Joachim H.; Pint, Bruce A.; Maziasz, Philip J.

    2007-05-01

    A corrosion resistant, electrically conductive component such as a bipolar plate for a PEM fuel cell includes 20 55% Cr, balance base metal such as Ni, Fe, or Co, the component having thereon a substantially external, continuous layer of chromium nitride.

  2. The seismotectonics of plate boundaries

    NASA Technical Reports Server (NTRS)

    Berger, J.; Brune, J. N.; Goodkind, J.; Wyatt, F.; Agnew, D. C.; Beaumont, C.

    1981-01-01

    Research on the seismotectonics of plate boundaries is summarized. Instrumental development and an observational program designed to study various aspects of the seismotectonics of southern California and the northern Gulf of California are described. A unique superconducting gravimeter was further developed and supported under this program for deployment and operation at several sites. Work on Earth tides is also discussed.

  3. Custom Multiwell Plate Design for Rapid Assembly of Photopatterned Hydrogels.

    PubMed

    Ahmed, Naveed; Schober, Joseph; Hill, Lindsay; Zustiak, Silviya P

    2016-06-01

    The extracellular matrix provides both mechanical support and biochemical cues that influence cellular behavior. Matrix stiffness, in particular, has been found to regulate cellular morphology, motility, proliferation, differentiation, and drug responses among other behaviors. Thus, biomaterial platforms that exhibit wide range of stiffness and are available in a semi high-throughput format such as a multiwell plate would be useful for elucidating cell-substrate relationships. Polyacrylamide (PA) gels have been widely used as cell platforms since they span a range of stiffness between 0.3 and 300 kPa in Young's modulus, which encompasses all soft tissues. However, PA gels are time consuming and labor intensive to prepare, and are not amenable to a multiwell plate format. In this study, we present a novel custom multiwell plate design that allows for a one-step stiffness assay assembly that reduces preparation time and labor intensity by several fold. Gel stiffness is controlled by ultraviolet light intensity and exposure time to achieve a wide stiffness range from a single gel precursor solution. The geometry of the gels is defined by a custom photomask and gel thickness is controlled by spacers. A multiwell plate upper structure is designed similar to a regular multiwell plate such that a gel fits in each well and cells and media are added on top. The upper structure design allows for adequate gas exchange and minimum evaporation. Comparison between cell behaviors seeded in the custom and a standard multiwell plate demonstrated the suitability of the design as a cell culture platform. In summary, we describe and validate a novel custom design for an easy and rapid assembly of photopolymerizable PA-based stiffness assay. PMID:27059131

  4. Avionics Box Cold Plate Damage Prevention

    NASA Technical Reports Server (NTRS)

    Stambolian, Damon B.; Larchar, Steven W.; Henderson, Gena; Tran, Donald; Barth, Tim

    2012-01-01

    Problem Introduction: 1. Prevent Cold Plate Damage in Space Shuttle. 1a. The number of cold plate problems had increased from an average of 16.5 per/year between 1990 through 2000, to an average of 39.6 per year between 2001through 2005. 1b. Each complete set of 80 cold plates cost approximately $29 million, an average of $362,500 per cold plate. 1c It takes four months to produce a single cold plate. 2. Prevent Cold Plate Damage in Future Space Vehicles.

  5. Consecutive plate acoustic suppressor apparatus and methods

    NASA Technical Reports Server (NTRS)

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

    1992-01-01

    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.

  6. 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.

  7. A robust electrochemiluminescence immunoassay for carcinoembryonic antigen detection based on a microtiter plate as a bridge and Au@Pd nanorods as a peroxidase mimic.

    PubMed

    Zhang, Yong; Pang, Xuehui; Wu, Dan; Ma, Hongmin; Yan, Zhaoqing; Zhang, Jiatao; Du, Bin; Wei, Qin

    2016-01-01

    The common drawbacks of most traditional electrochemiluminescence (ECL) immunoassays are the strict storage conditions for the ECL electrode and the steric hindrance caused by bovine serum albumin and antigen. The strict storage conditions require that the modified electrode must be stored at 4 °C before measurement, which may cause the degradation of protein molecules and low reproducibility as the time goes by. The steric hindrance can hinder electron transfer between the electrode and the electrochemical active substance unable to transmit proteins on the electrode surface. The current study takes a 96-well microtiter plate (MTP) as a bridge for analyte pre-treatment and Au@Pd nanorods as a peroxidase mimic to assemble a simple and robust ECL immunoassay. Advantages of such assay include not only high sensitivity but also robust detection circumstance. We demonstrated the method by detecting carcinoembryonic antigen from human serum and obtained a good detection limit of 3 fg mL(-1). PMID:26609799

  8. Cultural Neuroscience

    PubMed Central

    Ames, Daniel L.; Fiske, Susan T.

    2013-01-01

    Cultural neuroscience issues from the apparently incompatible combination of neuroscience and cultural psychology. A brief literature sampling suggests, instead, several preliminary topics that demonstrate proof of possibilities: cultural differences in both lower-level processes (e.g. perception, number representation) and higher-order processes (e.g. inferring others’ emotions, contemplating the self) are beginning to shed new light on both culture and cognition. Candidates for future cultural neuroscience research include cultural variations in the default (resting) network, which may be social; regulation and inhibition of feelings, thoughts, and actions; prejudice and dehumanization; and neural signatures of fundamental warmth and competence judgments. PMID:23874143

  9. Maps, Plates, and Mount Saint Helens.

    ERIC Educational Resources Information Center

    Lary, Barbara E.; Krockover, Gerald H.

    1987-01-01

    Describes a laboratory activity on plate tectonics which focuses on the connection between plate tectonics and the different types of volcanoes. Provides questions for discussion and includes suggestions for extending the activity. (ML)

  10. Plating by glass-bead peening

    NASA Technical Reports Server (NTRS)

    Babecki, A. J.; Haehner, C. L.

    1971-01-01

    Technique permits plating of primarily metallic substrates with either metals or nonmetals at normal temperature. Peening uses compressed air to apply concurrent streams of small glass beads and powdered plating material to the substrate.

  11. 49 CFR 230.46 - Badge plates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... TRANSPORTATION STEAM LOCOMOTIVE INSPECTION AND MAINTENANCE STANDARDS Boilers and Appurtenances Steam Gauges § 230.46 Badge plates. A metal badge plate showing the allowed steam pressure shall be attached to...

  12. 49 CFR 230.46 - Badge plates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... TRANSPORTATION STEAM LOCOMOTIVE INSPECTION AND MAINTENANCE STANDARDS Boilers and Appurtenances Steam Gauges § 230.46 Badge plates. A metal badge plate showing the allowed steam pressure shall be attached to...

  13. 49 CFR 230.46 - Badge plates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... TRANSPORTATION STEAM LOCOMOTIVE INSPECTION AND MAINTENANCE STANDARDS Boilers and Appurtenances Steam Gauges § 230.46 Badge plates. A metal badge plate showing the allowed steam pressure shall be attached to...

  14. 49 CFR 230.46 - Badge plates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... TRANSPORTATION STEAM LOCOMOTIVE INSPECTION AND MAINTENANCE STANDARDS Boilers and Appurtenances Steam Gauges § 230.46 Badge plates. A metal badge plate showing the allowed steam pressure shall be attached to...

  15. 49 CFR 230.46 - Badge plates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... TRANSPORTATION STEAM LOCOMOTIVE INSPECTION AND MAINTENANCE STANDARDS Boilers and Appurtenances Steam Gauges § 230.46 Badge plates. A metal badge plate showing the allowed steam pressure shall be attached to...

  16. 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.

  17. Electrically induced mechanical precompression of ferroelectric plates

    DOEpatents

    Chen, P.J.

    1987-03-02

    A method of electrically inducing mechanical precompression of ferroelectric plate covered with electrodes utilizes the change in strains of the plate as functions of applied electric field. A first field polarizes and laterally shrinks the entire plate. An outer portion of the electrodes are removed, and an opposite field partially depolarizes and expands the central portion of the plate against the shrunk outer portion. 2 figs.

  18. Practical automatic Arabic license plate recognition system

    NASA Astrophysics Data System (ADS)

    Mohammad, Khader; Agaian, Sos; Saleh, Hani

    2011-02-01

    Since 1970's, the need of an automatic license plate recognition system, sometimes referred as Automatic License Plate Recognition system, has been increasing. A license plate recognition system is an automatic system that is able to recognize a license plate number, extracted from image sensors. In specific, Automatic License Plate Recognition systems are being used in conjunction with various transportation systems in application areas such as law enforcement (e.g. speed limit enforcement) and commercial usages such as parking enforcement and automatic toll payment private and public entrances, border control, theft and vandalism control. Vehicle license plate recognition has been intensively studied in many countries. Due to the different types of license plates being used, the requirement of an automatic license plate recognition system is different for each country. [License plate detection using cluster run length smoothing algorithm ].Generally, an automatic license plate localization and recognition system is made up of three modules; license plate localization, character segmentation and optical character recognition modules. This paper presents an Arabic license plate recognition system that is insensitive to character size, font, shape and orientation with extremely high accuracy rate. The proposed system is based on a combination of enhancement, license plate localization, morphological processing, and feature vector extraction using the Haar transform. The performance of the system is fast due to classification of alphabet and numerals based on the license plate organization. Experimental results for license plates of two different Arab countries show an average of 99 % successful license plate localization and recognition in a total of more than 20 different images captured from a complex outdoor environment. The results run times takes less time compared to conventional and many states of art methods.

  19. Electrically induced mechanical precompression of ferroelectric plates

    DOEpatents

    Chen, Peter J.

    1987-01-01

    A method of electrically inducing mechanical precompression of a ferroelectric plate covered with electrodes utilizes the change in strains of the plate as functions of applied electric field. A first field polarizes and laterally shrinks the entire plate. An outer portion of the electrodes are removed, and an opposite field partially depolarizes and expands the central portion of the plate against the shrunk outer portion.

  20. 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.

  1. Ion plated electronic tube device

    DOEpatents

    Meek, T.T.

    1983-10-18

    An electronic tube and associated circuitry which is produced by ion plating techniques. The process is carried out in an automated process whereby both active and passive devices are produced at very low cost. The circuitry is extremely reliable and is capable of functioning in both high radiation and high temperature environments. The size of the electronic tubes produced are more than an order of magnitude smaller than conventional electronic tubes.

  2. Electroless metal plating of plastics

    DOEpatents

    Krause, L.J.

    1982-09-20

    Process for plating main group metals on aromatic polymers is carried out by the use of a nonaqueous solution of a salt of an alkali metal in a positive valence state and a main group metal in a negative valence state with contact between the solution and polymer providing a redox reaction causing the deposition of the main group metal and the reduction of the polymer. Products from the process exhibit useful decorative and electrical properties.

  3. Electroless metal plating of plastics

    DOEpatents

    Krause, Lawrence J.

    1986-01-01

    Process for plating main group metals on aromatic polymers is carried out by the use of a nonaqueous solution of a salt of an alkali metal in a positive valence state and a main group metal in a negative valence state with contact between the solution and polymer providing a redox reaction causing the deposition of the main group metal and the reduction of the polymer. Products from the process exhibit useful decorative and electrical properties.

  4. Electroless metal plating of plastics

    DOEpatents

    Krause, Lawrence J.

    1984-01-01

    Process for plating main group metals on aromatic polymers is carried out by the use of a nonaqueous solution of a salt of an alkali metal in a positive valence state and a main group metal in a negative valence state with contact between the solution and polymer providing a redox reaction causing the deposition of the main group metal and the reduction of the polymer. Products from the process exhibit useful decorative and electrical properties.

  5. PRETREATING URANIUM FOR METAL PLATING

    DOEpatents

    Wehrmann, R.F.

    1961-05-01

    A process is given for anodically treating the surface of uranium articles, prior to metal plating. The metal is electrolyzed in an aqueous solution of about 10% polycarboxylic acid, preferably oxalic acid, from 1 to 5% by weight of glycerine and from 1 to 5% by weight of hydrochloric acid at from 20 to 75 deg C for from 30 seconds to 15 minutes. A current density of from 60 to 100 amperes per square foot is used.

  6. Syncytial-Type Cell Plates

    PubMed Central

    Otegui, Marisa; Staehelin, L. Andrew

    2000-01-01

    Cell wall formation in the syncytial endosperm of Arabidopsis was studied by using high-pressure-frozen/freeze-substituted developing seeds and immunocytochemical techniques. The endosperm cellularization process begins at the late globular embryo stage with the synchronous organization of small clusters of oppositely oriented microtubules (∼10 microtubules in each set) into phragmoplast-like structures termed mini-phragmoplasts between both sister and nonsister nuclei. These mini-phragmoplasts produce a novel kind of cell plate, the syncytial-type cell plate, from Golgi-derived vesicles ∼63 nm in diameter, which fuse by way of hourglass-shaped intermediates into wide (∼45 nm in diameter) tubules. These wide tubules quickly become coated and surrounded by a ribosome-excluding matrix; as they grow, they branch and fuse with each other to form wide tubular networks. The mini-phragmoplasts formed between a given pair of nuclei produce aligned tubular networks that grow centrifugally until they merge into a coherent wide tubular network with the mini-phragmoplasts positioned along the network margins. The individual wide tubular networks expand laterally until they meet and eventually fuse with each other at the sites of the future cell corners. Transformation of the wide tubular networks into noncoated, thin (∼27 nm in diameter) tubular networks begins at multiple sites and coincides with the appearance of clathrin-coated budding structures. After fusion with the syncytial cell wall, the thin tubular networks are converted into fenestrated sheets and cell walls. Immunolabeling experiments show that the cell plates and cell walls of the endosperm differ from those of the embryo and maternal tissue in two features: their xyloglucans lack terminal fucose residues on the side chain, and callose persists in the cell walls after the cell plates fuse with the parental plasma membrane. The lack of terminal fucose residues on xyloglucans suggests that these cell wall

  7. Episodic plate tectonics on Venus

    NASA Technical Reports Server (NTRS)

    Turcotte, Donald

    1992-01-01

    Studies of impact craters on Venus from the Magellan images have placed important constraints on surface volcanism. Some 840 impact craters have been identified with diameters ranging from 2 to 280 km. Correlations of this impact flux with craters on the Moon, Earth, and Mars indicate a mean surface age of 0.5 +/- 0.3 Ga. Another important observation is that 52 percent of the craters are slightly fractured and only 4.5 percent are embayed by lava flows. These observations led researchers to hypothesize that a pervasive resurfacing event occurred about 500 m.y. ago and that relatively little surface volcanism has occurred since. Other researchers have pointed out that a global resurfacing event that ceased about 500 MYBP is consistent with the results given by a recent study. These authors carried out a series of numerical calculations of mantle convection in Venus yielding thermal evolution results. Their model considered crustal recycling and gave rapid planetary cooling. They, in fact, suggested that prior to 500 MYBP plate tectonics was active in Venus and since 500 MYBP the lithosphere has stabilized and only hot-spot volcanism has reached the surface. We propose an alternative hypothesis for the inferred cessation of surface volcanism on Venus. We hypothesize that plate tectonics on Venus is episodic. Periods of rapid plate tectonics result in high rates of subduction that cool the interior resulting in more sluggish mantle convection.

  8. Volcano spacing and plate rigidity

    SciTech Connect

    Brink, U. )

    1991-04-01

    In-plane stresses, which accompany the flexural deformation of the lithosphere under the load adjacent volcanoes, may govern the spacing of volcanoes in hotspot provinces. Specifically, compressive stresses in the vicinity of a volcano prevent new upwelling in this area, forcing a new volcano to develop at a minimum distance that is equal to the distance in which the radial stresses change from compressional to tensile (the inflection point). If a volcano is modeled as a point load on a thin elastic plate, then the distance to the inflection point is proportional to the thickness of the plate to the power of 3/4. Compilation of volcano spacing in seven volcanic groups in East Africa and seven volcanic groups of oceanic hotspots shows significant correlation with the elastic thickness of the plate and matches the calculated distance to the inflection point. In contrast, volcano spacing in island arcs and over subduction zones is fairly uniform and is much larger than predicted by the distance to the inflection point, reflecting differences in the geometry of the source and the upwelling areas.

  9. Plating of proximal humeral fractures.

    PubMed

    Martetschläger, Frank; Siebenlist, Sebastian; Weier, Michael; Sandmann, Gunther; Ahrens, Philipp; Braun, Karl; Elser, Florian; Stöckle, Ulrich; Freude, Thomas

    2012-11-01

    The optimal treatment for proximal humeral fractures is controversial. Few data exist concerning the influence of the surgical approach on the outcome. The purpose of this study was to evaluate the clinical and radiological outcomes of proximal humeral fractures treated with locking plate fixation through a deltopectoral vs an anterolateral deltoid-splitting approach. Of 86 patients who met the inclusion criteria, 70 were available for follow-up examination. Thirty-three patients were treated through a deltopectoral approach and 37 through an anterolateral deltoid-splitting approach. In all cases, open reduction and internal fixation with a PHILOS locking plate (Synthes, Umkirch, Germany) was performed. Clinical follow-up included evaluation of pain, shoulder mobility, and strength. Constant score and Disabilities of the Arm, Shoulder and Hand (DASH) score were assessed. A clinical neurological examination of the axillary nerve was also performed. Consolidation, reduction, and appearance of head necrosis were evaluated radiographically. After a mean follow-up of 33 months, Constant scores, DASH scores, and American Shoulder and Elbow Surgeons scores showed no significant differences between the groups. Clinical neurologic examination of the axillary nerve revealed no obvious damage to the nerve in either group. Deltopectoral and anterolateral detoid-splitting approaches for plate fixation of proximal humeral fractures are safe and provide similar clinical outcomes. The results of this study suggest that the approach can be chosen according to surgeon preference. PMID:23127451

  10. MTR plates modeling with MAIA

    SciTech Connect

    Marelle, V.; Dubois, S.; Ripert, M.; Noirot, J.

    2008-07-15

    MAIA is a thermo-mechanical code dedicated to the modeling of MTR fuel plates. The main physical phenomena modeled in the code are the cladding oxidation, the interaction between fuel and Al-matrix, the swelling due to fission products and the Al/fuel particles interaction. The creeping of the plate can be modeled in the mechanical calculation. MAIA has been validated on U-Mo dispersion fuel experiments such as IRIS 1 and 2 and FUTURE. The results are in rather good agreement with post-irradiation examinations. MAIA can also be used to calculate in-pile behavior of U{sub 3}Si{sub 2} plates as in the SHARE experiment irradiated in the SCK/Mol BR2 reactor. The main outputs given by MAIA throughout the irradiation are temperatures, cladding oxidation thickness, interaction thickness, volume fraction of meat constituents, swelling, displacements, strains and stresses. MAIA is originally a two-dimensional code but a three-dimensional version is currently under development. (author)

  11. 24 CFR 3280.5 - Data plate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 24 Housing and Urban Development 5 2014-04-01 2014-04-01 false Data plate. 3280.5 Section 3280.5... MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.5 Data plate. Each manufactured home shall bear a data plate affixed in a permanent manner near the main electrical panel or other...

  12. 24 CFR 3280.5 - Data plate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 24 Housing and Urban Development 5 2012-04-01 2012-04-01 false Data plate. 3280.5 Section 3280.5... MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.5 Data plate. Each manufactured home shall bear a data plate affixed in a permanent manner near the main electrical panel or other...

  13. 24 CFR 3280.5 - Data plate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 24 Housing and Urban Development 5 2011-04-01 2011-04-01 false Data plate. 3280.5 Section 3280.5... MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.5 Data plate. Each manufactured home shall bear a data plate affixed in a permanent manner near the main electrical panel or other...

  14. 24 CFR 3280.5 - Data plate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 24 Housing and Urban Development 5 2013-04-01 2013-04-01 false Data plate. 3280.5 Section 3280.5... MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.5 Data plate. Each manufactured home shall bear a data plate affixed in a permanent manner near the main electrical panel or other...

  15. 24 CFR 3280.5 - Data plate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 24 Housing and Urban Development 5 2010-04-01 2010-04-01 false Data plate. 3280.5 Section 3280.5... MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.5 Data plate. Each manufactured home shall bear a data plate affixed in a permanent manner near the main electrical panel or other...

  16. Moving Divertor Plates in a Tokamak

    SciTech Connect

    S.J. Zweben, H. Zhang

    2009-02-12

    Moving divertor plates could help solve some of the problems of the tokamak divertor through mechanical ingenuity rather than plasma physics. These plates would be passively heated on each pass through the tokamak and cooled and reprocessed outside the tokamak. There are many design options using varying plate shapes, orientations, motions, coatings, and compositions.

  17. Recent plate motions and crustal deformation

    SciTech Connect

    Lisowski, M. )

    1991-01-01

    Reports by U.S. workers on geodetic measurements of recent plate motions or crustal deformation published in 1987-1990 are reviewed. The review begins with global plate motions, proceeds through plate boundaries in California, Alaska, and the Pacific Northwest, and finishes with volcanic phenomena, monument stability and longevity, and GPS relative position measurements. 184 refs.

  18. 49 CFR 213.123 - Tie plates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Tie plates. 213.123 Section 213.123 Transportation... TRANSPORTATION TRACK SAFETY STANDARDS Track Structure § 213.123 Tie plates. (a) In Classes 3 through 5 track where timber crossties are in use there shall be tie plates under the running rails on at least eight...

  19. 46 CFR 169.665 - Name plates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Name plates. 169.665 Section 169.665 Shipping COAST... Gross Tons § 169.665 Name plates. Each generator, motor and other major item f power equipment must be provided with a name plate indicating the manufacturer's name, its rating in volts and amperes or in...

  20. 46 CFR 169.665 - Name plates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Name plates. 169.665 Section 169.665 Shipping COAST... Gross Tons § 169.665 Name plates. Each generator, motor and other major item f power equipment must be provided with a name plate indicating the manufacturer's name, its rating in volts and amperes or in...

  1. 46 CFR 169.665 - Name plates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Name plates. 169.665 Section 169.665 Shipping COAST... Gross Tons § 169.665 Name plates. Each generator, motor and other major item f power equipment must be provided with a name plate indicating the manufacturer's name, its rating in volts and amperes or in...

  2. 46 CFR 169.665 - Name plates.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Name plates. 169.665 Section 169.665 Shipping COAST... Gross Tons § 169.665 Name plates. Each generator, motor and other major item f power equipment must be provided with a name plate indicating the manufacturer's name, its rating in volts and amperes or in...

  3. 46 CFR 169.665 - Name plates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Name plates. 169.665 Section 169.665 Shipping COAST... Gross Tons § 169.665 Name plates. Each generator, motor and other major item f power equipment must be provided with a name plate indicating the manufacturer's name, its rating in volts and amperes or in...

  4. Forming Weld Lands On Metal Plates

    NASA Technical Reports Server (NTRS)

    Weddendorf, Bruce

    1994-01-01

    Forming shoe pounds edge of newly inserted plate workpiece. After many passes of shoe and advances of plate, thick land builds up at edge. Workpiece heated to enable metal to flow without strain hardening. Proposed upset-forming process replaces relatively expensive, time-consuming, and wasteful process in which integral weld lands created by machining metal away from plates everywhere except at lands.

  5. 21 CFR 872.4760 - Bone plate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Bone plate. 872.4760 Section 872.4760 Food and... DENTAL DEVICES Surgical Devices § 872.4760 Bone plate. (a) Identification. A bone plate is a metal device intended to stabilize fractured bone structures in the oral cavity. The bone segments are attached to...

  6. FASTENER FOR AN ASSEMBLY OF PLATES

    DOEpatents

    Groh, E.F.

    1963-08-20

    A fastener is provided for a spaced-apart parallel plate fuel assembly. The fastener, attached by screws to a key that passes through the edges of the assembled plate, serves as a retainer for the outermost plate as well as a bidirectional spacer for separating the fuel assembly from two neighboring fuel assemblies. (AEC)

  7. Normal-Pressure Tests of Rectangular Plates

    NASA Technical Reports Server (NTRS)

    Ramberg, Walter; Mcpherson, Albert E; Levy, Samuel

    1942-01-01

    Report presents the results of normal-pressure tests made of 56 rectangular plates with clamped edges and of 5 plates with freely supported edges. Pressure was applied and the center deflection and the permanent set at the center were measured. For some of the plates, in addition, strains and contours were measured.

  8. 21 CFR 872.4760 - Bone plate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Bone plate. 872.4760 Section 872.4760 Food and... DENTAL DEVICES Surgical Devices § 872.4760 Bone plate. (a) Identification. A bone plate is a metal device intended to stabilize fractured bone structures in the oral cavity. The bone segments are attached to...

  9. The critical shear load of rectangular plates

    NASA Technical Reports Server (NTRS)

    Seydel, Edgar

    1933-01-01

    This report gives formulas for analyzing the critical shear load of a simply supported square, isotropic (simple flat plate), or orthogonal anisotropic plate (a plate in which the rigidity in two directions perpendicular to each other is different, i.e. plywood or corrugated sheet), these formulas, although arrived at by approximation method, seem to agree fairly well with experimental results.

  10. Apparatus and method for controlling plating uniformity

    DOEpatents

    Hachman Jr., John T.; Kelly, James J.; West, Alan C.

    2004-10-12

    The use of an insulating shield for improving the current distribution in an electrochemical plating bath is disclosed. Numerical analysis is used to evaluate the influence of shield shape and position on plating uniformity. Simulation results are compared to experimental data for nickel deposition from a nickel--sulfamate bath. The shield is shown to improve the average current density at a plating surface.

  11. 49 CFR 451.23 - Plate specifications.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 6 2011-10-01 2011-10-01 false Plate specifications. 451.23 Section 451.23 Transportation Other Regulations Relating to Transportation (Continued) COAST GUARD, DEPARTMENT OF HOMELAND SECURITY SAFETY APPROVAL OF CARGO CONTAINERS TESTING AND APPROVAL OF CONTAINERS Safety Approval Plate § 451.23 Plate specifications. (a) The...

  12. Culturing Protozoa.

    ERIC Educational Resources Information Center

    Stevenson, Paul

    1980-01-01

    Compares various nutrient media, growth conditions, and stock solutions used in culturing protozoa. A hay infusion in Chalkey's solution maintained at a stable temperature is recommended for producing the most dense and diverse cultures. (WB)

  13. Repellent Culture.

    ERIC Educational Resources Information Center

    Carroll, Jeffrey

    2001-01-01

    Considers defining "culture," noting how it is difficult to define because those individuals defining it cannot separate themselves from it. Relates these issues to student writing and their writing improvement. Addresses violence in relation to culture. (SG)

  14. Liver Cell Culture Devices

    PubMed Central

    Andria, B.; Bracco, A.; Cirino, G.; Chamuleau, R. A. F. M.

    2010-01-01

    In the last 15 years many different liver cell culture devices, consisting of functional liver cells and artificial materials, have been developed. They have been devised for numerous different applications, such as temporary organ replacement (a bridge to liver transplantation or native liver regeneration) and as in vitro screening systems in the early stages of the drug development process, like assessing hepatotoxicity, hepatic drug metabolism, and induction/inhibition studies. Relevant literature is summarized about artificial human liver cell culture systems by scrutinizing PubMed from 2003 to 2009. Existing devices are divided in 2D configurations (e.g., static monolayer, sandwich, perfused cells, and flat plate) and 3D configurations (e.g., liver slices, spheroids, and different types of bioreactors). The essential features of an ideal liver cell culture system are discussed: different types of scaffolds, oxygenation systems, extracellular matrixes (natural and artificial), cocultures with nonparenchymal cells, and the role of shear stress problems. Finally, miniaturization and high-throughput systems are discussed. All these factors contribute in their own way to the viability and functionality of liver cells in culture. Depending on the aim for which they are designed, several good systems are available for predicting hepatotoxicity and hepatic metabolism within the general population. To predict hepatotoxicity in individual cases genomic analysis might be essential as well. PMID:26998397

  15. Culture Clubs.

    ERIC Educational Resources Information Center

    Gersten, Bridget Fitzgerald

    1998-01-01

    One way to break down barriers and promote understanding among English-as-a-Second-Language (ESL) and mainstream students is to establish culture clubs. Culture clubs involve frequent exchange of information about social, academic, and cultural topics in extracurricular settings. They are a critical component of ESL programs. The article explains…

  16. Teaching Culture.

    ERIC Educational Resources Information Center

    Magrath, Douglas R.

    The study of a foreign language is the study of another culture. Cultural involvement begins as learners progress from grammar to the actual use of language. Culture includes the ideas, customs, skills, arts, and tools of a people and influences both cognitive and affective behavior. It should be introduced as part of the total language…

  17. Development and testing of a multiwell plates absorbance reader for clinical analysis using inexpensive webcam

    NASA Astrophysics Data System (ADS)

    Castillo, Jimmy; Gutierrez, Hector; Vitta, Yosmery; Martinez, Mauro; Fernandez, Alberto

    2007-09-01

    Biochemical analysis and clinical tests like glucose, hemoglobin, cholesterol, iron, etc. are crucial for early illness diagnosis like diabetes, anemia and coronary deceases. These tests usually are done in state of the art instruments in well equipped laboratories in health centers. In some cases, these instruments are not portable, so they are not recommended for clinical field studies in remote areas. The present work shows a portable low-cost prototype of multi-well plates reader designed for clinical analysis. A Light Emission Diodes (LEDs) array is used as excitation source and an inexpensive webcam as detector. The light source illuminates the 96 well plates and the webcam take the image with 640x480 pixels. The data is acquired and processed by using a portable computer. 96 samples can be read including blanks and calibration standards simultaneously. Light absorption data are processed using a MatLab software designed in our laboratory to obtain calibration curves, standards lectures and samples concentration. The system was evaluated using different analytes series solutions: Neutral Red, Cooper (II) Ammonia Complex and Methyl Orange. The results shows that it is possible to measure few micro liters of solutions with adequate exactitude and precision of less than 3%. As possible analytical clinical application, iron determination was performed using Fe(III) Thiocyanate complex. This method is usually applied in serum samples analysis. The sensibility achieved with the proposed instrumentation configurations allows the analysis of iron in serum samples in the references values normal range (0.75 - 1.5 mg/L) in human.

  18. Plate tectonics, habitability and life

    NASA Astrophysics Data System (ADS)

    Spohn, Tilman; Breuer, Doris

    2016-04-01

    The role of plate tectonics in defining habitability of terrestrial planets is being increasingly discussed (e.g., Elkins-Tanton, 2015). Plate tectonics is a significantly evolved concept with a large variety of aspects. In the present context, cycling of material between near surface and mantle reservoirs is most important. But increased heat transport through mixing of cold lithosphere with the deep interior and formation of continental crust may also matter. An alternative mechanism of material cycling between these reservoirs is hot-spot volcanism combined with crust delamination. Hot-spot volcanism will transport volatiles to the atmosphere while delamination will mix crust, possibly altered by sedimentation and chemical reactions, with the mantle. The mechanism works as long as the stagnant lithosphere plate has not grown thicker than the crust and as long as volcanic material is added onto the crust. Thermal evolution studies suggest that the mechanism could work for the first 1-2 Ga of planetary evolution. The efficiency of the mechanism is limited by the ratio of extrusive to intrusive volcanism, which is thought to be less than 0.25. Plate tectonics would certainly have an advantage by working even for more evolved planets. A simple, most-used concept of habitability requires the thermodynamic stability of liquid water on the surface of a planet. Cycling of CO2between the atmosphere, oceans and interior through subduction and surface volcanism is an important element of the carbonate-silicate cycle, a thermostat feedback cycle that will keep the atmosphere from entering into a runaway greenhouse. Calculations for a model Earth lacking plate tectonics but degassing CO2, N, and H2O to form a surface ocean and a secondary atmosphere (Tosi et al, 2016) suggest that liquid water can be maintained on the surface for 4.5Ga. The model planet would then qualify as habitable. It is conceivable that the CO2 buffering capability of its ocean together with silicate

  19. MEPE is a novel regulator of growth plate cartilage mineralization

    PubMed Central

    Staines, K.A.; Mackenzie, N.C.W.; Clarkin, C.E.; Zelenchuk, L.; Rowe, P.S.; MacRae, V.E.; Farquharson, C.

    2012-01-01

    Matrix extracellular phosphoglycoprotein (MEPE) belongs to the SIBLING protein family which play key roles in biomineralization. Although the growth plates of MEPE-overexpressing mice display severe morphological disruption, the expression and function of MEPE in growth plate matrix mineralization remains largely undefined. Here we show MEPE and its cleavage product, the acidic serine aspartate-rich MEPE-associated motif (ASARM) peptide, to be localised to the hypertrophic zone of the growth plate. We also demonstrate that the phosphorylated (p)ASARM peptide inhibits ATDC5 chondrocyte matrix mineralization. Stable MEPE-overexpressing ATDC5 cells also had significantly reduced matrix mineralization in comparison to the control cells. Interestingly, we show that the addition of the non-phosphorylated (np)ASARM peptide promoted mineralization in the ATDC5 cells. The peptides and the overexpression of MEPE did not affect the differentiation of the ATDC5 cells. For a more physiologically relevant model, we utilized the metatarsal organ culture model. We show the pASARM peptide to inhibit mineralization at two stages of development, as shown by histological and μCT analysis. Like in the ATDC5 cells, the peptides did not affect the differentiation of the metatarsals indicating that the effects seen on mineralization are direct, as is additionally confirmed by no change in alkaline phosphatase activity or mRNA expression. In the metatarsal organ cultures, the pASARM peptide also reduced endothelial cell markers and vascular endothelial growth factor mRNA expression. Taken together these results show MEPE to be an important regulator of growth plate chondrocyte matrix mineralization through its cleavage to an ASARM peptide. PMID:22766095

  20. Peridinialean dinoflagellate plate patterns, labels and homologies

    USGS Publications Warehouse

    Edwards, L.E.

    1990-01-01

    Tabulation patterns for peridinialean dinoflagellate thecae and cysts have been traditionally expressed using a plate labelling system described by C.A. Kofoid in the early 1900's. This system can obscure dinoflagellate plate homologies and has not always been strictly applied. The plate-labelling system presented here introduces new series labels but incorporates key features and ideas from the more recently proposed systems of G.L. Eaton and F.J.R. Taylor, as modified by W.R. Evitt. Plate-series recognition begins with the cingulum (C-series) and proceeds from the cingulum toward the apex for the three series of the epitheca/epicyst and proceeds from the cingulum toward the antapex for the two series of the hypotheca/hypocyst. The epithecal/epicystal model consists of eight plates that touch the anterior margin of the cingulum (E-series: plates E1-E7, ES), seven plates toward the apex that touch the E-series plates (M-series: R, M1-M6), and up to seven plates near the apex that do not touch E-series plates (D-series: Dp-Dv). The hypothecal/hypocystal model consists of eight plates that touch the posterior margin of the cingulum (H-series: H1-H6,HR,HS) and three plates toward the antapex (T1-T3). Epithecal/epicystal tabulation patterns come in both 8- and 7- models, corresponding to eight and seven plates, respectively, in the E-series. Hypothecal/hypocystal tabulation patterns also come in both 8- and 7-models, corresponding to eight and seven plates, respectively, in the H-series. By convention, the 7-model epitheca/epicyst has no plates E1 and M1; the 7-model hypotheca/hypocyst has no plate H6. Within an 8-model or 7-model, the system emphasizes plates that are presumed to be homologous by giving them identical labels. I introduce the adjectives "monothigmate", "dithigmate," and "trithigmate" to designate plates touching one, two, and three plates, respectively, of the adjacent series. The term "thigmation" applies to the analysis of plate contacts between

  1. Development of flat-plate solar plate collector: Evaporator

    NASA Astrophysics Data System (ADS)

    Abramzon, B.; Yaron, I.

    1981-11-01

    In the present study the thermal performance of a flat plate solar collector is analyzed theoretically for the case in which the working fluid may undergo a phase change within the tubes of the collector. In addition to the common domestic applications, such a collector - evaporator may be used as a generator of vapors for the production of mechanical or electrical energy, e.g., solar water pumps, solar power stations, etc., as well as for solar - powered absorption refrigeration machines, distillation installations, etc.

  2. Separator plate for a fuel cell

    DOEpatents

    Petri, R.J.; Meek, J.; Bachta, R.P.; Marianowski, L.G.

    1996-04-02

    A separator plate is described for a fuel cell comprising an anode current collector, a cathode current collector and a main plate, the main plate disposed between the anode current collector and the cathode current collector. The anode current collector forms a flattened peripheral wet seal structure and manifold wet seal structure on the anode side of the separator plate and the cathode current collector forms a flattened peripheral wet seal structure and manifold wet seal structure on the cathode side of the separator plate. In this manner, the number of components required to manufacture and assemble a fuel cell stack is reduced. 9 figs.

  3. Separator plate for a fuel cell

    DOEpatents

    Petri, Randy J.; Meek, John; Bachta, Robert P.; Marianowski, Leonard G.

    1996-01-01

    A separator plate for a fuel cell comprising an anode current collector, a cathode current collector and a main plate, the main plate disposed between the anode current collector and the cathode current collector. The anode current collector forms a flattened peripheral wet seal structure and manifold wet seal structure on the anode side of the separator plate and the cathode current collector forms a flattened peripheral wet seal structure and manifold wet seal structure on the cathode side of the separator plate. In this manner, the number of components required to manufacture and assemble a fuel cell stack is reduced.

  4. Steady state response of unsymmetrically laminated plates

    SciTech Connect

    Hosokawa, Kenji; Kawashima, Katsuya; Sakata, Toshiyuki

    1995-11-01

    A numerical approach for analyzing the forced vibration problem of a symmetrically laminated FRP (fiber reinforced plastic) composite plate was proposed by the authors. In the present paper, this approach is modified for application to an unsymmetrically laminated FRP composite plate. Numerical calculations are carried out for the clamped antisymmetrically laminated rectangular and elliptical plates which are a kind of unsymmetrically laminated plate. Then,, the effects of the lamina material and the fiber orientation angle on the steady state response are discussed. Furthermore, it is investigated that what structural damping factor is most influenced on the steady state response of an antisymmetrically laminated plate.

  5. Detection of archaeological forgeries of Iberian lead plates using nanoelectrochemical techniques. The lot of fake plates from Bugarra (Spain).

    PubMed

    Doménech-Carbó, Antonio; Doménech-Carbó, María Teresa; Pérez, Monserrat Lastras; Herrero-Cortell, Miquel

    2015-02-01

    Identification of forgeries is of considerable interest in studies of archaeological signariums and written Iberian artifacts, elements of capital importance for the knowledge of that culture, because there are many Iberian inscribed lead plate counterfeits circulating in the market and among many museum funds. A case study of identification of forgeries of archaeological lead using voltammetry of microparticles (VMP) and scanning electrochemical microscopy (SECM), aided by conventional optical microscopy and scanning electron microscopy (SEM/EDX) is described. The electrochemical methods are essentially non-invasive so can be applied to samples of nanoscopic size. Application to the authentication of five lead plates found in the archaeological site of Los Villaricos-Torralba (Bugarra, Spain) is described. PMID:25549575

  6. Shear deformation in thick auxetic plates

    NASA Astrophysics Data System (ADS)

    Lim, Teik-Cheng

    2013-08-01

    This paper aims to understand the effect of auxeticity on shear deformation in thick plates. Three models for the shear correction factor of plates as a function of Poisson’s ratio were proposed: an analytical model, a cubic fit model and a modified model. Of these three, the cubic fit model exhibits the best accuracy over the entire range of Poisson’s ratio from -1 to 0.5. The extent of shear deformation is herein investigated using the example of uniformly loaded circular plates. It was found that the maximum deformation of such plates based on Mindlin theory approximates to those according to Kirchhoff theory when the Poisson’s ratio of the plate material is highly negative. When the Poisson’s ratio of the plate material is -1 and the edge of the plate is simply supported, the calculation of the maximum deflection by Mindlin theory simplifies into that by Kirchhoff theory. These results suggest that auxeticity reduces shear deformation in thick plates, permitting the use of classical plate theory for thick plates only if the plate material is highly auxetic.

  7. Characterization of gas-liquid mass transfer phenomena in microtiter plates.

    PubMed

    Hermann, Robert; Lehmann, Mathias; Büchs, Jochen

    2003-01-20

    Gas-liquid mass transfer properties of shaken 96-well microtiter plates were characterized using a recently described method. The maximum oxygen transfer capacity (OTR(max)), the specific mass transfer area (a), and the mass transfer coefficient (k(L)) in a single well were determined at different shaking intensities (different shaking frequencies and shaking diameters at constant filling volume) and different filling volumes by means of sulfite oxidation as a chemical model system. The shape (round and square cross-sections) and the size (up to 2 mL maximum filling volume) of a microtiter plate well were also considered as influencing parameters. To get an indication of the hydrodynamic behavior of the liquid phase in a well, images were taken during shaking and the liquid height derived as a characteristic parameter. The investigations revealed that the OTR(max) is predominantly dependent on the specific mass transfer area (a) for the considered conditions in round-shaped wells. The mass transfer coefficient (k(L)) in round-shaped wells remains at a nearly constant value of about 0.2 m/h for all shaking intensities, thus within the range reported in the literature for surface-aerated bioreactors. The OTR(max) in round-shaped wells is strongly influenced by the interfacial tension, determined by the surface tension of the medium used and the surface properties of the well material. Up to a specific shaking intensity the liquid surface in the wells remains horizontal and no liquid movement can be observed. This critical shaking intensity must be exceeded to overcome the surface tension and, thus, to increase the liquid height and enlarge the specific mass transfer area. This behavior is solely specific to microtiter plates and has not yet been observed for larger shaking bioreactors such as shaking flasks. In square-shaped microtiter plate wells the corners act as baffles and cause a significant increase of OTR(max), a, and k(L). An OTR(max) of up to 0.15 mol

  8. An understanding of HSLA-65 plate steels

    NASA Astrophysics Data System (ADS)

    Sampath, K.

    2006-02-01

    HSLA-65 plate steels can be produced using one of five plate manufacturing techniques: normalizing, controlled rolling (CR), controlled rolling followed by accelerated cooling (CR-AC), direct quenching and tempering (DQT), or conventional quenching and tempering (Q&T). The HSLA-65 steels are characterized by low carbon content and low alloy content, and they exhibit a low carbon equivalent that allows improved plate weldability. These characteristics in turn (a) provide the steel plate with a refined microstructure that ensures high strength and toughness; (b) eliminate or substantially reduce the need for preheating during welding; (c) resist susceptibility to hydrogen-assisted cracking (HAC) in the weld heat affected zone (HAZ) when fusion (arc) welded using low heat-input conditions; and (d) depending on section thickness, facilitate high heat-input welding (about 2 kJ/mm) without significant loss of strength or toughness in the HAZ. However, application of this plate manufacturing process and of these controls produces significant differences in the metallurgical structure and range of mechanical properties of the HSLA-65 plate steels both among themselves and versus conventional higher strength steel (HSS) plates. For example, among the HSLA-65 plate steels, those produced by Q&T exhibit minimal variability in mechanical properties, especially in thicker plates. Besides variability in mechanical properties depending on plate thickness, the CR and CR-AC plate steels exhibit a relatively higher yield strength to ultimate tensile strength (YS/UTS) ratio than do DQT and Q&T steels. Such differences in processing and properties of HSLA-65 plate steels could potentially affect the selection and control of various secondary fabrication practices, including arc welding. Consequently, fabricators must exercise extreme caution when transferring allowable limits of certified secondary fabrication practices from one type of HSLA-65 plate steel to another, even for the

  9. When Did Plate Tectonics Begin

    NASA Astrophysics Data System (ADS)

    Brown, M.

    2015-12-01

    Present-day plate tectonics on Earth is characterized by asymmetric (one-sided) subduction, but how do we recognize the imprint of subduction in the geologic record? How do we weigh global (commonly younger) vs local (commonly older) datasets or distinguish initiation from episodic from continuous subduction? How reliable are data gaps? Characteristics of the Paleozoic record of subduction include calc-alkaline magmatism, blueschist/UHP metamorphism and collisional orogenesis, and ophiolites as representatives of former ocean lithosphere. Are these characteristic rocks preserved in Proterozoic, Archean and Hadean crust? Does a hotter mantle, higher heat production and weaker lithosphere modify or eliminate these features? What preceded subduction and how do we recognize that regime? Are rock associations or geochemical fingerprints reliable? Does reworking and overprinting modify geochemical fingerprints? Proposals for the start of plate tectonics have been based on: persistence of isotope anomalies/fractionated chemical domains in the mantle; changes in chemistry of magmatic rocks, rates of crustal growth vs reworking, and sites of growth; the metamorphic record, particularly the first appearance of contrasting thermal gradients or eclogite (including evidence from mineral inclusions in diamonds) or UHP metamorphic rocks; stabilization of cratonic lithosphere and formation of supercratons, and the beginning of the Proterozoic supercontinent cycle; the end of the flat Earth, emergence of continents, development of significant topography, changes in the style of orogeny and the rise in atmospheric oxygen; and, the appearance of passive margins and changes in the style of sedimentation. Estimates of the timing have varied from the Hadean to Neoproterozoic. I will summarize evidence for a growing consensus that the late Mesoarchean to early Paleoproterozoic was a 700 Myr long period of transition to continuous (?) subduction and global (?) mobile-lid plate tectonics.

  10. Tectonic speed limits from plate kinematic reconstructions

    NASA Astrophysics Data System (ADS)

    Zahirovic, Sabin; Müller, R. Dietmar; Seton, Maria; Flament, Nicolas

    2015-05-01

    The motion of plates and continents on the planet's surface are a manifestation of long-term mantle convection and plate tectonics. Present-day plate velocities provide a snapshot of this ongoing process, and have been used to infer controlling factors on the speeds of plates and continents. However, present-day velocities do not capture plate behaviour over geologically representative periods of time. To address this shortcoming, we use a plate tectonic reconstruction approach to extract time-dependent plate velocities and geometries from which root mean square (RMS) velocities are computed, resulting in a median RMS plate speed of ∼ 4 cm /yr over 200 Myr. Linking tectonothermal ages of continental lithosphere to the RMS plate velocity analysis, we find that the increasing portions of plate area composed of continental and/or cratonic lithosphere significantly reduces plate speeds. Plates with any cratonic portion have a median RMS velocity of ∼ 5.8 cm /yr, while plates with more than 25% of cratonic area have a median RMS speed of ∼ 2.8 cm /yr. The fastest plates (∼ 8.5 cm /yr RMS speed) have little continental fraction and tend to be bounded by subduction zones, while the slowest plates (∼ 2.6- 2.8 cm /yr RMS speed) have large continental fractions and usually have little to no subducting part of plate perimeter. More generally, oceanic plates tend to move 2-3 times faster than continental plates, consistent with predictions of numerical models of mantle convection. The slower motion of continental plates is compatible with deep keels impinging on asthenospheric flow and increasing shear traction, thus anchoring the plate in the more viscous mantle transition zone. We also find that short-lived (up to ∼ 10 Myr) rapid accelerations of Africa (∼100 and 65 Ma), North America (∼100 and 55 Ma) and India (∼ 130 , 80 and 65 Ma) appear to be correlated with plume head arrivals as recorded by large igneous province (LIPs) emplacement. By evaluating

  11. Turbulent flows near flat plates

    NASA Astrophysics Data System (ADS)

    Kambe, R.; Imamura, T.; Doi, M.

    1980-08-01

    The method to study the effect of the plate moving in the homogeneous or isotropic turbulence is presented. The crucial point of this method is to solve the Orr-Sommerfeld like equation, which is satisfied by the kernel of the Wiener-Hermite expansion of the velocity field, under the inhomogeneous boundary condition. In the special case of constant mean flow, our method gives the same result as that of Hunt and Graham and succeeds in explaining the experimental results of Thomas and Hancock. The method is also applied to the case of nonuniform mean flow, where the shear effect comes up.

  12. Non-cyanide silver plating

    SciTech Connect

    Dini, J.W.

    1995-11-07

    Lawrence Livermore National Laboratory (LLNL) and Technic, Inc. have entered into a CRADA (Cooperative Research and Development Agreement) with the goal of providing industry with an environmentally benign alternative to the presently used silver cyanide plating process. This project has been in place for about six months and results are quite promising. The main objective, that of deposition of deposits as thick as 125 um (5 mils), has been met. Property data such as stress and hardness have been obtained and the structure of the deposit has been analyzed via metallography and x-ray diffraction. These results will be presented in this paper, along with plans for future work.

  13. Coarse Layering at 'Home Plate'

    NASA Technical Reports Server (NTRS)

    2006-01-01

    This image shows coarse-grained layers from around the edge of a low plateau called 'Home Plate' inside Mars' Gusev Crater. One possible origin is material falling to the ground after being thrown aloft by an explosion such as a volcanic eruption or meteorite impact.

    The panoramic camera (Pancam) on NASA's Mars Exploration Rover Spirit acquired the exposures for this image on Spirit's 749th Martian day (Feb. 10, 2006). This view is an approximately true-color rendering mathematically generated from separate images taken through all of the left Pancam's 432-nanometer to 753-nanometer filters.

  14. Cultured Inquiry

    ERIC Educational Resources Information Center

    Scheppler, Judith A.; Sethakorn, Nan; Styer, Susan

    2003-01-01

    The Kirby-Bauer assay, also called the disc diffusion assay, is a standard procedure used in clinical laboratories to test the susceptibility of patients' bacterial isolates to antibiotics. In the assay, the bacteria are swabbed onto an agar plate, and paper discs impregnated with antibiotics are placed on the agar. The antibiotic diffuses from…

  15. Adherent protective coatings plated on magnesium-lithium alloy

    NASA Technical Reports Server (NTRS)

    1965-01-01

    Zinc is plated on a magnesium-lithium alloy by using a modification of the standard zinc-plate immersion bath. Further protection is given the alloy by applying a light plating of copper on the zinc plating. Other metals are plated on the copper by using conventional plating baths.

  16. A Refined Shear Deformation Plate Theory

    NASA Astrophysics Data System (ADS)

    Liu, Yucheng

    2011-04-01

    An improved higher-order shear deformation theory of plates is presented in this paper. The theory is developed from the transverse shear deformation theory presented by Ambartsumian [11]. The present plate theory contains kinematics of higher-order displacement field of plates, a system of higher-order differential equilibrium equations in terms of the three generalized displacements of bending plates, and a system of boundary conditions at each edge of plate boundaries. The present shear deformation theory of plates is validated by applying it to solve torsional plates and simply supported plates. The obtained solutions using the present theory are compared with the solutions of other shear-deformation theories. A good agreement is achieved through these comparisons and the advantages of the present theory are clearly verified. The shear deformation plate theory presented here can be applied to the analysis of laminated composite plates to better predict their dynamic and static behaviors. The proposed theory should also be supplemented to the theory of finite element analysis for developing new shell elements.

  17. Analysis of plate spin motion and its implications for strength of plate boundary

    NASA Astrophysics Data System (ADS)

    Matsuyama, Takeshi; Iwamori, Hikaru

    2016-02-01

    In this study, we investigate the driving forces of plate motion, especially those of plate spin motion, that are related to the toroidal components of the global plate velocity field. In previous works, numerical simulations of mantle convection have been used to examine the extent to which toroidal velocity components are naturally generated on the surface, by varying key parameters, notably the rheological properties of plates and plate boundaries. Here, we take the reverse approach and perform analyses of observed plate motions, which show an increase in the toroidal/poloidal ratio at high degrees of spherical harmonic expansion, as well as a rapid change in the plate spin rate and the estimated driving stress around a critical plate size of approximately 1000 km. This quantitative relationship constrains the strength at plate boundaries to 3-75 MPa, which is consistent with several seismological observations, including those from the NE Japan arc associated with the 2011 Tohoku earthquake.

  18. Statistical Evaluation of Electronic and Plate Counts for Estimating Bacterial Populations

    PubMed Central

    Mountney, G. J.; O'malley, J.

    1966-01-01

    Bacteria from cultures of Staphylococcus aureus, Pseudomonas geniculata, and Rhodotorula glutenis were counted by use of an electronic counter and by plate counts from broth cultures at selected periods from 0 to 24 hr. Variations in the two methods were noted, and the results were compared after calculating correlations, coefficients of variation, and nested analyses of variance. It was not possible to determine the absolute accuracy of the two methods; however, the precision of the results obtained with an electronic counter was better than that obtained with plate counting. It appears that electronic counting, because of ease and convenience, provides better comparative counts at a particular stage of growth, especially during the early stages of the growth period, whereas plate counting yields the best results in determining growth curves. PMID:16349683

  19. Wide-Field Plate Database

    NASA Astrophysics Data System (ADS)

    Tsvetkov, M. K.; Stavrev, K. Y.; Tsvetkova, K. P.; Semkov, E. H.; Mutatov, A. S.

    The Wide-Field Plate Database (WFPDB) and the possibilities for its application as a research tool in observational astronomy are presented. Currently the WFPDB comprises the descriptive data for 400 000 archival wide field photographic plates obtained with 77 instruments, from a total of 1 850 000 photographs stored in 269 astronomical archives all over the world since the end of last century. The WFPDB is already accessible for the astronomical community, now only in batch mode through user requests sent by e-mail. We are working on on-line interactive access to the data via INTERNET from Sofia and parallel from the Centre de Donnees Astronomiques de Strasbourg. (Initial information can be found on World Wide Web homepage URL http://www.wfpa.acad.bg.) The WFPDB may be useful in studies of a variety of astronomical objects and phenomena, andespecially for long-term investigations of variable objects and for multi-wavelength research. We have analysed the data in the WFPDB in order to derive the overall characteristics of the totality of wide-field observations, such as the sky coverage, the distributions by observation time and date, by spectral band, and by object type. We have also examined the totality of wide-field observations from point of view of their quality, availability and digitisation. The usefulness of the WFPDB is demonstrated by the results of identification and investigation of the photometrical behaviour of optical analogues of gamma-ray bursts.

  20. Petrologic implications of plate tectonics.

    PubMed

    Yoder, H S

    1971-07-30

    Petrologists can make significant contributions to the plate tectonic concept. Fixing the stability fields of the principal rock types involved will provide the limits of pressure and temperature of the various environments. Experimental determination of the partition coefficients of the trace elements will be helpful. Studies of the partial melting behavior of possible parental materials in the absence and presence of water, especially the undersaturated region, will contribute to the understanding of magma production. Experimental observations on the rheological properties of the peridotites below and just above the solidus will lead to a better evaluation of the convective mechanism. Measurement of the fundamental properties of rocks, such as the density of solids and liquids at high pressures and temperatures, would contribute to understanding the concepts of diapiric rise, magma segregation, and the low-velocity zone. Broader rock sampling of the oceanic areas of all environments will do much to define the petrologic provinces. The field petrologist specializing in the Paleozoic regions and Precambrian shields can contribute by examining those regions for old plate boundaries and devising new criteria for their recognition. PMID:17770454

  1. Skin or nail culture

    MedlinePlus

    Mucosal culture; Culture - skin; Culture - mucosal; Nail culture; Culture - fingernail; Fingernail culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria, ...

  2. Current global plate kinematics from GPS (1995-2007) with the plate-consistent reference frame

    NASA Astrophysics Data System (ADS)

    Kogan, Mikhail G.; Steblov, Grigory M.

    2008-04-01

    We present the vectors of rotation of 10 major lithospheric plates, estimated from continuous GPS observations at 192 globally distributed stations; 71 stations were selected as representing stable plate regions. All days for the period 1995.0-2007.0 were included in the analysis. In contrast to previous GPS plate models, our model is independent of international terrestrial reference frames (ITRF). The origin of our plate-consistent reference frame is the center of plate rotation (CP) rather than the center of mass of the entire Earth's system (CM) as in recent versions of ITRF. We estimate plate rotations and CP by minimizing the misfit between the horizontal velocities predicted by the plate model and the observed GPS velocities. If any version of ITRF is used as the reference frame, the drift of the ITRF origin relative to CP cannot be neglected in estimation of plate rotation vectors and plate-residual station velocities. The model of the plate kinematics presented here addresses the problem debated since the beginning of the space geodesy: how big are disagreements between the current plate motions and the motions averaged over several million years? We compare the vectors of relative plate rotations estimated here with the published vectors from GPS and geologic models. We also discuss the integrity of individual plates as exhibited by plate-residual station velocities. For seven largest plates, the RMS value of plate-residual station velocities in stable plate interiors is 0.5-0.9 mm/a; this value is an upper bound on deviation of real plates from infinite stiffness.

  3. Scale-down of vinegar production into microtiter plates using a custom-made lid.

    PubMed

    Schlepütz, Tino; Büchs, Jochen

    2014-04-01

    As an important food preservative and condiment, vinegar is widely produced in industry by submerged acetic acid bacteria cultures. Although vinegar production is established on the large scale, up to now suitable microscale cultivation methods, e.g. using microtiter plates, are missing to enable high-throughput cultivation and to optimize fermentation conditions. In order to minimize evaporation losses of ethanol and acetic acid in a 48-well microtiter plate during vinegar production a new custom-made lid was developed. A diffusion model was used to calculate the dimensions of a hole in the lid to guarantee a suitable oxygen supply and level of ventilation. Reference fermentation was conducted in a 9-L bioreactor to enable the calculation of the proper cultivation conditions in the microtiter plate. The minimum dissolved oxygen tensions in the microtiter plate were between 7.5% and 23% of air saturation and in the same range as in the 9-L bioreactor. Evaporation losses of ethanol and acetic acid were less than 5% after 47 h and considerably reduced compared to those of microtiter plate fermentations with a conventional gas-permeable seal. Furthermore, cultivation times in the microtiter plate were with about 40 h as long as in the 9-L bioreactor. In conclusion, microtiter plate cultivations with the new custom-made lid provide a platform for high-throughput studies on vinegar production. Results are comparable to those in the 9-L bioreactor. PMID:24315857

  4. Effect of plate roughness on the field near RPC plates

    NASA Astrophysics Data System (ADS)

    Jash, A.; Majumdar, N.; Mukhopadhyay, S.; Chattopadhyay, S.

    2016-06-01

    The inner surfaces of the electrodes encompassing the gas volume of a Resistive Plate Chamber (RPC) have been found to exhibit asperities with, grossly, three kind of features. The desired uniform electric field within the gas volume of RPC is expected to be affected due to the presence of these asperities, which will eventually affect the final response from the detector. In this work, an attempt has been made to model the highly complex roughness of the electrode surfaces and compute its effect on the electrostatic field within RPC gas chamber. The calculations have been performed numerically using Finite Element Method (FEM) and Boundary Element Method (BEM) and the two methods have been compared in this context.

  5. Droplet-based magnetic bead immunoassay using microchannel-connected multiwell plates (μCHAMPs) for the detection of amyloid beta oligomers.

    PubMed

    Park, Min Cheol; Kim, Moojong; Lim, Gun Taek; Kang, Sung Min; An, Seong Soo A; Kim, Tae Song; Kang, Ji Yoon

    2016-06-21

    Multiwell plates are regularly used in analytical research and clinical diagnosis but often require laborious washing steps and large sample or reagent volumes (typically, 100 μL per well). To overcome such drawbacks in the conventional multiwell plate, we present a novel microchannel-connected multiwell plate (μCHAMP) that can be used for automated disease biomarker detection in a small sample volume by performing droplet-based magnetic bead immunoassay inside the plate. In this μCHAMP-based immunoassay platform, small volumes (30-50 μL) of aqueous-phase working droplets are stably confined within each well by the simple microchannel structure (200-300 μm in height and 0.5-1 mm in width), and magnetic beads are exclusively transported into an adjacent droplet through the oil-filled microchannels assisted by a magnet array aligned beneath and controlled by a XY-motorized stage. Using this μCHAMP-based platform, we were able to perform parallel detection of synthetic amyloid beta (Aβ) oligomers as a model analyte for the early diagnosis of Alzheimer's disease (AD). This platform easily simplified the laborious and consumptive immunoassay procedure by achieving automated parallel immunoassay (32 assays per operation in 3-well connected 96-well plate) within 1 hour and at low sample consumption (less than 10 μL per assay) with no cumbersome manual washing step. Moreover, it could detect synthetic Aβ oligomers even below 10 pg mL(-1) concentration with a calculated detection limit of ∼3 pg mL(-1). Therefore, the μCHAMP and droplet-based magnetic bead immunoassay, with the combination of XY-motorized magnet array, would be a useful platform in the diagnosis of human disease, including AD, which requires low consumption of the patient's body fluid sample and automation of the entire immunoassay procedure for high processing capacity. PMID:27185215

  6. Assay of creatine kinase in microtiter plates using thio-NAD to allow monitoring at 405 nM.

    PubMed

    Florini, J R

    1989-11-01

    An assay system for creatine kinase using microtiter plates and a plate reader that records absorbancies at 405 nM has been devised. The system is an adaptation of well-established assays that couple creatine kinase with the reactions catalyzed by hexokinase and glucose-6-phosphate dehydrogenase (G6PDH), to give a measurable increase in reduced pyridine nucleotide quantitated by absorbance at 340 nM. Two features of this system are modified for reading at 405 nM: (i) The thioamido derivative of NAD is used because its reduced form exhibits a substantial increase in absorbance at 405 nM, the most commonly available wavelength on microplate readers; and (ii) glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides is used because it can reduce either NAD or NADP (unlike most other G6PDH enzymes, which require NADP), thus making it unnecessary to use the more expensive thio-NADP. The rate of thio-NAD reduction is linear with enzyme concentration and time over a 20-fold range of concentrations of purified creatine kinase, and the assay also works well with myogenic cells allowed to grow and differentiate in the 96-well plate in which the assay is performed. This system offers considerable savings in cells, time, and material in studies of muscle cell differentiation, for which creatine kinase levels are frequently measured. It also provides a potential method for the convenient and economical measurement of activities of many other enzymes that can be coupled to reduction of thio-NAD. PMID:2610356

  7. Battery plate containing filler with conductive coating

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1986-01-01

    The plate (10) comprises a matrix or binder resin phase (12) in which is dispersed particulate, conductive tin oxide such as tin oxide coated glass fibers (14). A monopolar plate (11) is prepared by coating a layer (18) of electrolytically active material onto a surface of the plate (10). Tin oxide is prevented from reduction by coating a surface of the plate (10) with a conductive, impervious layer resistant to reduction such as a thin film (22) of lead adhered to the plate with a layer (21) of conductive adhesive. The plate (10) can be formed by casting a molten dispersion from mixer (36) onto a sheet (30) of lead foil or by passing an assembly of a sheet (41) of resin, a sheet (43) of fiberglass and a sheet (45) of lead between the nip of heated rollers (48, 50).

  8. Battery plate containing filler with conductive coating

    NASA Technical Reports Server (NTRS)

    Rowlette, John J. (Inventor)

    1985-01-01

    The plate (10) comprises a matrix or binder resin phase (12) in which is dispersed particulate, conductive tin oxide such as tin oxide coated glass fibers (14). A monopolar plate (11) is prepared by coating a layer (18) of electrolytically active material onto a surface of the plate (10). Tin oxide is prevented from reduction by coating a surface of the plate (10) with a conductive, impervious layer resistant to reduction such as a thin film (22) of lead adhered to the plate with a layer (21) of conductive adhesive. The plate (10) can be formed by casting a molten dispersion from mixer (36) onto a sheet (30) of lead foil or by passing an assembly of a sheet (41) of resin, a sheet (43) of fiberglass and a sheet (45) of lead between the nip of heated rollers (48, 50).

  9. Excitation and Characterization of Chladni Plate Patterns

    NASA Astrophysics Data System (ADS)

    Bourke, Shannon; Behringer, Ernest

    2011-04-01

    When a thin metal plate with a small amount of sand on it is made to vibrate, aesthetically pleasing sand patterns can form along the nodal lines of the plate. These symmetric patterns are called Chladni Patterns. Students taking PHY 101 Physical Science in the Arts at Eastern Michigan University create these patterns by pulling a violin bow across the edge of a plate, or by using a mechanical oscillator to drive the center of a plate. These two methods only allow a small subset of all possible points on the plate to be excited. We designed and built an electronic device that allows its user to excite the plate at any point. We present patterns created with this electronic device and other methods, and describe ways to model the observed patterns.

  10. Membrane- and plate-type acoustic metamaterials.

    PubMed

    Huang, Tai-Yun; Shen, Chen; Jing, Yun

    2016-06-01

    Over the past decade there has been a great amount of research effort devoted to the topic of acoustic metamaterials (AMMs). The recent development of AMMs has enlightened the way of manipulating sound waves. Several potential applications such as low-frequency noise reduction, cloaking, angular filtering, subwavelength imaging, and energy tunneling have been proposed and implemented by the so-called membrane- or plate-type AMMs. This paper aims to offer a thorough overview on the recent development of membrane- or plate-type AMMs. The underlying mechanism of these types of AMMs for tuning the effective density will be examined first. Four different groups of membrane- or plate-type AMMs (membranes with masses attached, plates with masses attached, membranes or plates without masses attached, and active AMMs) will be reviewed. The opportunities, limitations, and challenges of membrane- or plate-type AMMs will be also discussed. PMID:27369148

  11. Rapidly Moving Divertor Plates In A Tokamak

    SciTech Connect

    S. Zweben

    2011-05-16

    It may be possible to replace conventional actively cooled tokamak divertor plates with a set of rapidly moving, passively cooled divertor plates on rails. These plates would absorb the plasma heat flux with their thermal inertia for ~10-30 sec, and would then be removed from the vessel for processing. When outside the tokamak, these plates could be cooled, cleaned, recoated, inspected, and then returned to the vessel in an automated loop. This scheme could provide nearoptimal divertor surfaces at all times, and avoid the need to stop machine operation for repair of damaged or eroded plates. We describe various possible divertor plate designs and access geometries, and discuss an initial design for a movable and removable divertor module for NSTX-U.

  12. Coupling between plate vibration and acoustic radiation

    NASA Technical Reports Server (NTRS)

    Frendi, Abdelkader; Maestrello, Lucio; Bayliss, Alvin

    1992-01-01

    A detailed numerical investigation of the coupling between the vibration of a flexible plate and the acoustic radiation is performed. The nonlinear Euler equations are used to describe the acoustic fluid while the nonlinear plate equation is used to describe the plate vibration. Linear, nonlinear, and quasi-periodic or chaotic vibrations and the resultant acoustic radiation are analyzed. We find that for the linear plate response, acoustic coupling is negligible. However, for the nonlinear and chaotic responses, acoustic coupling has a significant effect on the vibration level as the loading increases. The radiated pressure from a plate undergoing nonlinear or chaotic vibrations is found to propagate nonlinearly into the far-field. However, the nonlinearity due to wave propagation is much weaker than that due to the plate vibrations. As the acoustic wave propagates into the far-field, the relative difference in level between the fundamental and its harmonics and subharmonics decreases with distance.

  13. Real time monitoring of electroless nickel plating

    NASA Astrophysics Data System (ADS)

    Rains, Aaron E.; Kline, Ronald A.

    2013-01-01

    This work deals with the design and manufacturing of the heat and chemical resistant transducer case required for on-line immersion testing, experimental design, data acquisition and signal processing. Results are presented for several depositions with an accuracy of two ten-thousandths of an inch in coating thickness obtained. Monitoring the deposition rate of Electroless Nickel (EN) plating in-situ will provide measurement of the accurate dimensions of the component being plated, in real time. EN is used as for corrosion and wear protection for automotive an - Electroless Nickel (EN) plating is commonly used for corrosion and wear protection for automotive and aerospace components. It plates evenly and symmetrically, theoretically allowing the part to be plated to its final dimension. Currently the standard approach to monitoring the thickness of the deposited nickel is to remove the component from the plating bath and physically measure the part. This can lead to plating problems such as pitting, non-adhesion of the deposit and contamination of the plating solution. The goal of this research effort is to demonstrate that plating thickness can be rapidly and accurately measured using ultrasonic testing. Here a special housing is designed to allow immersion of the ultrasonic transducers directly into the plating bath. An FFT based signal processing algorithm was developed to resolve closely spaced echoes for precise thickness determination. The technique in this research effort was found to be capable of measuring plating thicknesses to within 0.0002 inches. It is expected that this approach will lead to cost savings in many EN plating operations.

  14. High speed thin plate fatigue crack monitor

    NASA Technical Reports Server (NTRS)

    Wincheski, Buzz A. (Inventor); Heyman, Joseph S. (Inventor); Namkung, Min (Inventor); Fulton, James P. (Inventor)

    1996-01-01

    A device and method are provided which non-destructively detect crack length and crack geometry in thin metallic plates. A non-contacting vibration apparatus produces resonant vibrations without introducing extraneous noise. Resulting resonant vibration shifts in cracked plates are correlated to known crack length in plates with similar resonant vibration shifts. In addition, acoustic emissions of cracks at resonance frequencies are correlated to acoustic emissions from known crack geometries.

  15. Avionics Box Cold Plate Damage Prevention

    NASA Technical Reports Server (NTRS)

    Stambolian, Damon; Larcher, Steven; Henderson, Gena; Tran, Donald

    2011-01-01

    Over the years there have been several occurrences of damage to Space Shuttle Orbiter cold plates during removal and replacement of avionics boxes. Thus a process improvement team was put together to determine ways to prevent these kinds of damage. From this effort there were many solutions including, protective covers, training, and improved operations instructions. The focus of this paper is to explain the cold plate damage problem and the corrective actions for preventing future damage to aerospace avionics cold plate designs.

  16. Anisotropic magnetocapacitance in ferromagnetic-plate capacitors

    NASA Astrophysics Data System (ADS)

    Haigh, J. A.; Ciccarelli, C.; Betz, A. C.; Irvine, A.; Novák, V.; Jungwirth, T.; Wunderlich, J.

    2015-04-01

    The capacitance of a parallel-plate capacitor can depend on the applied magnetic field. Previous studies have identified capacitance changes induced via classical Lorentz force or spin-dependent Zeeman effects. Here we measure a magnetization direction-dependent capacitance in parallel-plate capacitors where one plate is a ferromagnetic semiconductor, gallium manganese arsenide. This anisotropic magnetocapacitance is due to the anisotropy in the density of states dependent on the magnetization through the strong spin-orbit interaction.

  17. Hyperspectral imaging for detecting pathogens grown on agar plates

    NASA Astrophysics Data System (ADS)

    Yoon, Seung Chul; Lawrence, Kurt C.; Siragusa, Gregory R.; Line, John E.; Park, Bosoon; Windham, William R.

    2007-09-01

    This paper is concerned with the development of a hyperspectral imaging technique for detecting and identifying one of the most common foodborne pathogens, Campylobacter. Direct plating using agars is an effective tool for laboratory tests and analyses of microorganisms. The morphology (size, growth pattern, color, etc.) of colonies grown on agar plates has been widely used to tentatively differentiate organisms. However, it is sometimes difficult to differentiate target organisms like Campylobacters from other contaminants grown together on the same agar plates. A hyperspectral imaging system operating at the visible and near infrared (VNIR) spectral region from 400 nm to 900 nm was set up to measure spectral signatures of 17 different Campylobacter and non-Campylobacter subspecies. Protocols for culturing, imaging samples and for calibrating measured data were developed. The VNIR spectral library of all 17 organisms commonly encountered in poultry was established from calibrated hyperspectral images. A classification algorithm was developed to locate and identify Campylobacters, non-Campylobacter contaminants, and background agars with 99.29% accuracy. This research has a potential to be expanded to detect other pathogens grown on agar media.

  18. Bipolar plates for PEM fuel cells

    NASA Astrophysics Data System (ADS)

    Middelman, E.; Kout, W.; Vogelaar, B.; Lenssen, J.; de Waal, E.

    The bipolar plates are in weight and volume the major part of the PEM fuel cell stack, and are also a significant contributor to the stack costs. The bipolar plate is therefore a key component if power density has to increase and costs must come down. Three cell plate technologies are expected to reach targeted cost price levels, all having specific advantages and drawbacks. NedStack has developed a conductive composite materials and a production process for fuel cell plates (bipolar and mono-polar). The material has a high electric and thermal conductivity, and can be processed into bipolar plates by a proprietary molding process. Process cycle time has been reduced to less than 10 s, making the material and process suitable for economical mass production. Other development work to increase material efficiency resulted in thin bipolar plates with integrated cooling channels, and integrated seals, and in two-component bipolar plates. Total thickness of the bipolar plates is now less than 3 mm, and will be reduced to 2 mm in the near future. With these thin integrated plates it is possible to increase power density up to 2 kW/l and 2 kW/kg, while at the same time reducing cost by integrating other functions and less material use.

  19. Plate Tectonics: A Paradigm under Threat.

    ERIC Educational Resources Information Center

    Pratt, David

    2000-01-01

    Discusses the challenges confronting plate tectonics. Presents evidence that contradicts continental drift, seafloor spreading, and subduction. Reviews problems posed by vertical tectonic movements. (Contains 242 references.) (DDR)

  20. Vibration of thin, tensioned, helically wrapped plates

    NASA Astrophysics Data System (ADS)

    Lopez, Ernesto; Müftü, Sinan

    2011-02-01

    Free vibration analysis of a thin tensioned plate, wrapped around a cylindrical guide in a helical manner is presented. The system is a model of a thin, flexible web wrapped around a turn-bar. The equation of motion of the wrapped plate is derived by using the energy method and with the Kirchhoff-Love assumptions. The weak form of the equation of motion was obtained by the finite element method and the eigenvalue problem was solved numerically. The effects of parameters such as plate tension, guide radius, longitudinal and helical wrap angles, plate width, and the lengths of the non-wrapped segments were investigated. Eigenmodes with same mode numbers were observed in symmetric and anti-symmetric fashion about the center of the plate, for symmetrically wrapped plates. It was shown that the plate/shell boundary of the wrapped plate effectively acts like a support. For non-helically wrapped plates the free edges cause a frequency clustering of the lateral modes about the dominant longitudinal mode. The frequency clustering diminishes when helical wrap is introduced.

  1. The floor plate: multiple cells, multiple signals.

    PubMed

    Placzek, Marysia; Briscoe, James

    2005-03-01

    One of the key organizers in the CNS is the floor plate - a group of cells that is responsible for instructing neural cells to acquire distinctive fates, and that has an important role in establishing the elaborate neuronal networks that underlie the function of the brain and spinal cord. In recent years, considerable controversy has arisen over the mechanism by which floor plate cells form. Here, we describe recent evidence that indicates that discrete populations of floor plate cells, with characteristic molecular properties, form in different regions of the neuraxis, and we discuss data that imply that the mode of floor plate induction varies along the anteroposterior axis. PMID:15738958

  2. Dynamic stiffness formulation for free orthotropic plates

    NASA Astrophysics Data System (ADS)

    Ghorbel, O.; Casimir, J. B.; Hammami, L.; Tawfiq, I.; Haddar, M.

    2015-06-01

    This paper presents a procedure for developing the dynamic stiffness matrix of a free orthotropic Kirchhoff plate. The dynamic stiffness matrix is computed for free edge boundary conditions of the plate that allow assembly procedures. The method is based on a strong formulation of Kirchhoff plate equations and series solutions, taking advantage of the symmetry and Gorman type decomposition of the free boundary conditions. The performances of the so-called Dynamic Stiffness Method (DSM) are evaluated by comparing the harmonic responses of an orthotropic Kirchhoff plate with those obtained from the Finite Element Method using four noded quadrilateral elements.

  3. Locomotion of a flapping flexible plate

    NASA Astrophysics Data System (ADS)

    Hua, Ru-Nan; Zhu, Luoding; Lu, Xi-Yun

    2013-12-01

    The locomotion of a flapping flexible plate in a viscous incompressible stationary fluid is numerically studied by an immersed boundary-lattice Boltzmann method for the fluid and a finite element method for the plate. When the leading-edge of the flexible plate is forced to heave sinusoidally, the entire plate starts to move freely as a result of the fluid-structure interaction. Mechanisms underlying the dynamics of the plate are elucidated. Three distinct states of the plate motion are identified and can be described as forward, backward, and irregular. Which state to occur depends mainly on the heaving amplitude and the bending rigidity of the plate. In the forward motion regime, analysis of the dynamic behaviors of the flapping flexible plate indicates that a suitable degree of flexibility can improve the propulsive performance. Moreover, there exist two kinds of vortex streets in the downstream of the plate which are normal and deflected wake. Further the forward motion is compared with the flapping-based locomotion of swimming and flying animals. The results obtained in the present study are found to be consistent with the relevant observations and measurements and can provide some physical insights into the understanding of the propulsive mechanisms of swimming and flying animals.

  4. Rapidly solidified metal coatings by peen plating

    NASA Technical Reports Server (NTRS)

    Chu, H. P.

    1987-01-01

    Specimens of 7075-T6 aluminum alloy were peen plated with rapidly solidified tin-lead and aluminum powders, and the cross-sections of the coated specimens were examined by light and electron microscopy. The properties of the peen plated specimens were also compared with those of shot peened specimens without any coating. It is found that peen plating with rapidly solidified metals improves the fatigue properties of the coated samples to a greater extent than shot peening alone. Specimens of 7075-T6 alloy peen plated with rapidly solidified tin-lead and aluminum exhibited better fatigue resistance than shot peened specimens in both air and salt water.

  5. MACMA: a Virtual Lab for Plate Tectonics

    NASA Astrophysics Data System (ADS)

    Grigne, C.; Combes, M.; Tisseau, C.

    2013-12-01

    MACMA (Multi-Agent Convective MAntle) is a tool developed to simulate evolutive plate tectonics and mantle convection in a 2-D cylindrical geometry (Combes et al., 2012). The model relies mainly on a force balance to compute the velocity of each plate, and on empirical rules to determine how plate boundaries move and evolve. It includes first-order features of plate tectonics: (a) all plates on Earth do not have the same size, (b) subduction zones are asymmetric, (c) plates driven by subducting slabs and upper plates do not exhibit the same velocities, and (d) plate boundaries are mobile, can collide, merge and disappear, and new plate boundaries can be created. The MACMA interface was designed to be user-friendly and a simple use of the simulator can be achieved without any prerequisite knowledge in fluid dynamics, mantle rheology, nor in numerical methods. As a preliminary study, the simulator was used by a few students from bachelor's degree to master's degree levels. An initial configuration for plate tectonics has to be created before starting a simulation: the number and types of plate boundaries (ridge, subduction, passive margins) has to be defined and seafloor ages must be given. A simple but interesting exercise consists in letting students build such an initial configuration: they must analyze a map of tectonic plates, choose a 2-D section and examine carefully a map of seafloor ages. Students mentioned that the exercise made them realize that the 3-D spherical structure of plate tectonics does not translate directly in a simple 2-D section, as opposed to what is usually shown in books. Physical parameters: e.g. mantle viscosity, number of layers to consider in the mantle (upper and lower mantle, possible asthenosphere), initial time and mantle temperature, have to be chosen, and students can use this virtual lab to see how different scenarios emerge when parameters are varied. Very importantly, the direct visualization of the mobility of plate

  6. Fuel cell cooler-humidifier plate

    DOEpatents

    Vitale, Nicholas G.; Jones, Daniel O.

    2000-01-01

    A cooler-humidifier plate for use in a proton exchange membrane (PEM) fuel cell stack assembly is provided. The cooler-humidifier plate combines functions of cooling and humidification within the fuel cell stack assembly, thereby providing a more compact structure, simpler manifolding, and reduced reject heat from the fuel cell. Coolant on the cooler side of the plate removes heat generated within the fuel cell assembly. Heat is also removed by the humidifier side of the plate for use in evaporating the humidification water. On the humidifier side of the plate, evaporating water humidifies reactant gas flowing over a moistened wick. After exiting the humidifier side of the plate, humidified reactant gas provides needed moisture to the proton exchange membranes used in the fuel cell stack assembly. The invention also provides a fuel cell plate that maximizes structural support within the fuel cell by ensuring that the ribs that form the boundaries of channels on one side of the plate have ends at locations that substantially correspond to the locations of ribs on the opposite side of the plate.

  7. Wave propagation in metamaterial lattice sandwich plates

    NASA Astrophysics Data System (ADS)

    Fang, Xin; Wen, Jihong; Yin, Jianfei; Yu, Dianlong

    2016-04-01

    This paper designed a special acoustic metamaterial 3D Kagome lattice sandwich plate. Dispersion properties and vibration responses of both traditional plate and metamaterial plate are investigated based on FEA methods. The traditional plate does not have low-frequency complete bandgaps, but the metamaterial plate has low-frequency complete bandgap (at 620Hz) coming from the symmetrical local cantilever resonators. The bandgap frequency is approximate to the first-order natural frequency of the oscillator. Complex wave modes are analyzed. The dispersion curves of longitudinal waves exist in the flexural bandgap. The dispersion properties demonstrate the metamaterial design is advantageous to suppress the low-frequency flexural wave propagation in lattice sandwich plate. The flexural vibrations near the bandgap are also suppressed efficiently. The longitudinal excitation stimulates mainly longitudinal waves and lots of low-frequency flexural vibration modes are avoided. Furthermore, the free edge effects in metamaterial plate provide new method for damping optimizations. The influences of damping on vibrations of the metamaterial sandwich plate are studied. Damping has global influence on the wave propagation; stronger damping will induce more vibration attenuation. The results enlighten us damping and metamaterial design approaches can be unite in the sandwich plates to suppress the wave propagations.

  8. Modal interaction in postbuckled plates. Theory

    NASA Technical Reports Server (NTRS)

    Thurston, Gaylen A.

    1989-01-01

    Plates can have more than one buckled solution for a fixed set of boundary conditions. The theory for the identification and the computation of multiple solutions in buckled plates is examined. The theory predicts modal interaction (which is also called change in buckle pattern or secondary buckling) in experiments on certain plates with multiple theoretical solutions. A set of coordinate functions is defined for Galerkin's method so that the von Karman plate equations are reduced to a coupled set of cubic equations in generalized coordinates that are uncoupled in the linear terms. An iterative procedure for solving modal interaction problems is suggested based on this cubic form.

  9. Cultural Communications.

    ERIC Educational Resources Information Center

    Armas, Jose

    It is too often taken for granted that the communication process with culturally different children takes place as readily as it might with children from Anglo cultures. Most teachers receive training in verbal and formal communication skills; children come to school with nonverbal and informal communication skills. This initially can create…

  10. Ryukyuan Culture.

    ERIC Educational Resources Information Center

    Trafton, Terry

    The Ryukyu Islands of Japan, of which Okinawa is the best known, possess a lengthy history and a sophisticated cultural background, an exploration of which helps to shed light on this area and on mainland Japan. This document is an exposition of Ryukuan culture. Divided into eight sections, the areas covered include: (1) Historical perspective;…

  11. Bronchoscopic culture

    MedlinePlus

    ... a laboratory exam to check a piece of tissue or fluid from the lungs for infection-causing germs. ... Culture - bronchoscopic ... used to get a sample ( biopsy ) of lung tissue or fluid. The sample ... a special dish (culture). It is then watched to see if bacteria ...

  12. Tunable surface plasmon wave plates.

    PubMed

    Djalalian-Assl, Amir; Cadusch, Jasper J; Balaur, Eugeniu; Aramesh, Morteza

    2016-07-01

    The highest resonant transmission through an array of holes perforated in metallic screens occurs when the dielectric constant of the substrate, the superstrate, and the hole are the same. Changes in the refractive index of the homogenous environment also produce the largest shift in resonances per refractive index unit. In this Letter, we first propose and apply a technique in realization of a freestanding bi-periodic array of holes perforated in a silver film. We then show both numerically and experimentally that shifts in (1,0) and (0,1) modes in response to changes in the refractive index of the surrounding dielectric provide a mechanism for realization of a miniaturized tunable quarter-wave plate that operates in an extraordinary optical transmission mode with a high throughput and a near unity state of circularly polarized light. PMID:27367123

  13. Background events in microchannel plates

    NASA Technical Reports Server (NTRS)

    Siegmund, O. H. W.; Vallerga, J.; Wargelin, B.

    1988-01-01

    Measurements have been made to assess the characteristics and origins of background events in microchannel plates (MCPs). An overall background rate of about 0.4 events/sq cm persec has been achieved consistently for MCPs that have been baked and scrubbed. The temperature and gain of the MCPs are found to have no significant effect on the background rate. Detection of 1.46-MeV gamma rays from the MCP glass confirms the presence of K-40, with a concentration of 0.0007 percent, in MCP glass. It is shown that beta decay from K-40 is sufficient to cause the background rate and spectrum observed. Anticoincidence measurements indicate the the background rate caused by cosmic ray interactions is small (less than 0.016 events/sq cm per sec).

  14. A Treatise on Equivalent-Plate Stiffnesses for Stiffened Laminated-Composite Plates and Plate-Like Lattices

    NASA Technical Reports Server (NTRS)

    Nemeth, Michael P.

    2011-01-01

    A survey of studies conducted since 1914 on the use of equivalent-plate stiffnesses in modeling the overall, stiffness-critical response of stiffened plates and shells is presented. Two detailed, comprehensive derivations of first-approximation equivalent-plate stiffnesses are also presented that are based on the Reissner-Mindlin-type, first-order transverse-shear deformation theory for anisotropic plates. Equivalent-plate stiffness expressions, and a corresponding symbolic manipulation computer program, are also presented for several different stiffener configurations. These expressions are very general and exhibit the full range of anisotropies permitted by the Reissner-Mindlin-type, first-order transverse-shear deformation theory for anisotropic plates. The expressions presented in the present study were also compared with available, previously published results. For the most part, the previously published results are for special cases of the general expressions presented herein and are almost in complete agreement. Analysis is also presented that extends the use of the equivalent-plate stiffness expressions to sandwich plates.

  15. Thickness dependence of the Casimir force between a magnetodielectric plate and a diamagnetic plate

    SciTech Connect

    Inui, Norio

    2011-11-15

    This paper examines the repulsive Casimir force between a magnetodielectric plate, with static permeability greater than static permittivity, and a diamagnetic plate. As the thickness of the magnetodielectric plate is decreased, the attractive component of the Casimir force decreases more than the repulsive one. This effect makes the net Casimir force repulsive, and a larger repulsive Casimir force is generated compared to the Casimir force between the plates with infinite thickness.

  16. Recovery of Clostridia on Catalase-Treated Plating Media

    PubMed Central

    Harmon, Stanley M.; Kautter, Donald A.

    1977-01-01

    Four plating media commonly used for culturing clostridia were tested for their ability to support growth of several Clostridium species after storage of the plates for 1 to 10 days at 4 and 25°C with and without subsequent addition of catalase. Liver-veal (LV) agar and brain heart infusion (BHI) agar rapidly became incapable of supporting growth after storage without added catalase, whereas Shahidi Ferguson perfringens agar base and Brewer anaerobic agar were less affected. Plate counts of vegetative cells of nine of the less fastidious Clostridium species on untreated LV and BHI agars, stored for 3 days at 4°C, were 60 to 90% lower than counts on catalase-treated media. Counts on Shahidi Ferguson perfringens agar base were only 1 to 24% lower on untreated medium with the same species. Addition of 500 U of purified beef liver catalase to the surface of the 3-day-old agars before inoculation resulted in substantial restoration of the ability of the media to support colony formation from vegetative cells except with the most strictly anaerobic species (nonproteolytic C. botulinum types B, E, and F, and C. novyii types A and B). A similar response was obtained with spores of the less fastidious species on catalase-treated media. Our results suggest that inhibition of most Clostridium species on LV and BHI agars may be due to accumulation of peroxide during preparation, storage, and incubation of the media, and also suggest that the presence of glucose in these media is a major factor contributing to their inability to support growth. It is believed that the addition of exogenous catalase prevents the accumulation of peroxide(s), thus allowing colony formation from vegetative cells of the clostridia under what would otherwise be unsuitable cultural conditions. PMID:869526

  17. Cultural History and Cultural Materialism.

    ERIC Educational Resources Information Center

    Berman, Ronald

    1990-01-01

    Historicism critiques cultural history and cultural materialism as a methodology for literary analysis. Questions the finality of interpretation, how original values change, and whether dramatic history implies actual history. Using Shakespearean plays, analyzes the power and politics of a play in relation to its audience; posits that cultural…

  18. Evaluation of the 3M Petrifilm Enterobacteriaceae Count plate method for the enumeration of Enterobacteriaceae in foods.

    PubMed

    Silbernagel, Karen M; Lindberg, Kathryn G

    2002-09-01

    Results of the 3M Petrifilm Enterobacteriaceae Count (EB) plate method were compared with those of the standard violet red bile glucose agar (VRBG) method for the detection and enumeration of Enterobacteriaceae. Studies involving 107 bacterial strains demonstrated that the Petrifilm EB plate method is as sensitive as and more selective than the VRBG method. Sixty of the 62 pure Enterobacteriaceae cultures were recovered by both methods. In addition, 38 of the 45 non-Enterobacteriaceae organisms did not grow on the Petrifilm EB plate, while 28 of the 45 non-Enterobacteriaceae organisms did not grow on the VRBG plate. Colony counts from 174 naturally contaminated and 120 artificially inoculated dairy and nondairy food samples showed that the Petrifilm EB plate method performed as well as or better than the standard VRBG method for the enumeration of Enterobacteriaceae. PMID:12233857

  19. Evaluation of brush plated alloys as substitutes for tank plated hard chromium

    SciTech Connect

    Durham, H.B.; Hooper, A.M.

    1995-11-01

    This paper summarizes results obtained from a test program conducted in cooperation with Tinker Air Force Base (TAFB) in 1993 for the purpose of evaluating the potential of using brush plated alloys as replacements for tank plated hard chromium which is used in the overhaul of jet engines. Tank plating is energy expensive and generates waste products in several of the plating steps. Test specimens used in this study were fabricated from carbon steel, chromium-based stainless steel, and nickel-based stainless steel. Baseline specimens were tank plated with hard chromium or with a soft nickel capped with hard chromium. The specimens were tested for fatigue, thickness, microhardness, Taber wear, and Falex pin and vee block wear. All test were conducted in accordance with standard procedures of the American Society for Testing and Materials (ASTM). Test results obtained from the baseline specimens were compared with those obtained from specimens which had been coated with the three brush plated layered alloy solutions used in this evaluation. Results of this study show that brush plated alloy coatings appear promising as alternatives to tank plated hard chromium. Some data quality problems were encountered during testing, so at this time, definitive statements concerning the use of brush plating alloys as an acceptable alternative to tank plated hard chromium in critical TAFB maintenance, cannot be made. Although these tests results are not positive for TAFB`s operation, users of hard chromium tank plating with less critical applications may find brush plated coatings a suitable alternative. 1 ref.

  20. Simulation of Evolutive Plate Tectonics: the Size of Plates Depends on Mantle Temperature

    NASA Astrophysics Data System (ADS)

    Grigne, C.; Combes, M.

    2013-12-01

    We use a dynamic model of plate tectonics based on a multiagent approach, in a 2D cylindrical geometry (Combes et al., 2012), to study how evolutive plate tectonics affect the long term thermal state of the mantle, and in return, to analyze the relationship between the mantle mean temperature and the geometry of plate tectonics. Our model accounts for first-order features of plate tectonics: (a) all plates on Earth do not have the same size, (b) subduction zones are asymmetric, (c) plates driven by subducting slabs and upper plates do not exhibit the same velocities, and (d) plate boundaries are mobile, can collide, merge and disappear, and new plate boundaries can be created. We show that when processes for plate boundary creation (subduction initiation and ridge creation) are relying on a brittle criterion, namely when a fixed yield strength has to be reached, the average size of plates adapts to the mantle thermal state: longer plates are obtained for a hotter mantle, which implies a maximum seafloor age that remains fairly high throughout Earth's thermal history and limits mantle heat loss. This is consistent with petrological and geochemical constraints on Earth's cooling history. Important fluctuations in the mantle heat flux and velocities of plates are obtained on a timescale of a few hundred Myr, but on the long term, the relationship between the average wavelength of plate tectonics and mantle temperature can be explained by a simple scaling law. Recent compilations of geological records infer that passive margins had longer lifespans in the past (e.g. Bradley 2008; 2011), which has been linked to 'sluggish' plate tectonics and slow plates in the Precambrian (Korenaga, 2006). Our simulations outputs include lifespans of tectonic entities such as passive margins, as well as statistical data about events of plates reorganizations. We obtain faster plates in the past than at present day, but counterintuitively we also observe a low episodicity of tectonic

  1. Vacuum fusion bonding of glass plates

    DOEpatents

    Swierkowski, Steve P.; Davidson, James C.; Balch, Joseph W.

    2000-01-01

    An improved apparatus and method for vacuum fusion bonding of large, patterned glass plates. One or both glass plates are patterned with etched features such as microstructure capillaries and a vacuum pumpout moat, with one plate having at least one hole therethrough for communication with a vacuum pumpout fixture. High accuracy alignment of the plates is accomplished by a temporary clamping fixture until the start of the fusion bonding heat cycle. A complete, void-free fusion bond of seamless, full-strength quality is obtained through the plates; because the glass is heated well into its softening point and because of a large, distributed force that is developed that presses the two plates together from the difference in pressure between the furnace ambient (high pressure) and the channeling and microstructures in the plates (low pressure) due to the vacuum drawn. The apparatus and method may be used to fabricate microcapillary arrays for chemical electrophoresis; for example, any apparatus using a network of microfluidic channels embedded between plates of glass or similar moderate melting point substrates with a gradual softening point curve, or for assembly of glass-based substrates onto larger substrates, such as in flat panel display systems.

  2. Vacuum fusion bonding of glass plates

    DOEpatents

    Swierkowski, Steve P.; Davidson, James C.; Balch, Joseph W.

    2001-01-01

    An improved apparatus and method for vacuum fusion bonding of large, patterned glass plates. One or both glass plates are patterned with etched features such as microstructure capillaries and a vacuum pumpout moat, with one plate having at least one hole therethrough for communication with a vacuum pumpout fixture. High accuracy alignment of the plates is accomplished by a temporary clamping fixture until the start of the fusion bonding heat cycle. A complete, void-free fusion bond of seamless, full-strength quality is obtained through the plates; because the glass is heated well into its softening point and because of a large, distributed force that is developed that presses the two plates together from the difference in pressure between the furnace ambient (high pressure) and the channeling and microstructures in the plates (low pressure) due to the vacuum drawn. The apparatus and method may be used to fabricate microcapillary arrays for chemical electrophoresis; for example, any apparatus using a network of microfluidic channels embedded between plates of glass or similar moderate melting point substrates with a gradual softening point curve, or for assembly of glass-based substrates onto larger substrates, such as in flat panel display systems.

  3. Pressure-Letdown Plates for Coal Gasifiers

    NASA Technical Reports Server (NTRS)

    Collins, E. R., Jr.

    1985-01-01

    Variation of pseudoporous plates used with coal gasifiers in pressure letdown stage of processing minimize clogging. Rotating plates containing variable gap annuli continually change flow path to enable erosionless reduction of gas pressure. Particles that otherwise clog porous plugs pass through gaps.

  4. Computing relative plate velocities: a primer

    SciTech Connect

    Bevis, M.

    1987-08-01

    Standard models of present-day plate motions are framed in terms of rates and poles of rotation, in accordance with the well-known theorem due to Euler. This article shows how computation of relative plate velocities from such models can be viewed as a simple problem in spherical trigonometry. A FORTRAN subroutine is provided to perform the necessary computations.

  5. Plate Tectonic Cycle. K-6 Science Curriculum.

    ERIC Educational Resources Information Center

    Blueford, J. R.; And Others

    Plate Tectonics Cycle is one of the units of a K-6 unified science curriculum program. The unit consists of four organizing sub-themes: (1) volcanoes (covering formation, distribution, and major volcanic groups); (2) earthquakes (with investigations on wave movements, seismograms and sub-suface earth currents); (3) plate tectonics (providing maps…

  6. 49 CFR 236.779 - Plate, top.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 4 2011-10-01 2011-10-01 false Plate, top. 236.779 Section 236.779 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF... OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Definitions § 236.779 Plate, top....

  7. 49 CFR 236.779 - Plate, top.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 4 2014-10-01 2014-10-01 false Plate, top. 236.779 Section 236.779 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF... OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Definitions § 236.779 Plate, top....

  8. 49 CFR 236.779 - Plate, top.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Plate, top. 236.779 Section 236.779 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF... OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Definitions § 236.779 Plate, top....

  9. 49 CFR 236.779 - Plate, top.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Plate, top. 236.779 Section 236.779 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF... OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Definitions § 236.779 Plate, top....

  10. 49 CFR 236.779 - Plate, top.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 4 2013-10-01 2013-10-01 false Plate, top. 236.779 Section 236.779 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF... OF SIGNAL AND TRAIN CONTROL SYSTEMS, DEVICES, AND APPLIANCES Definitions § 236.779 Plate, top....

  11. Improved nickel plating of Inconel X-750

    NASA Technical Reports Server (NTRS)

    Farmer, M. E.; Feeney, J. E.; Kuster, C. A.

    1969-01-01

    Electroplating technique with acid pickling provides a method of applying nickel plating on Inconel X-750 tubing to serve as a wetting agent during brazing. Low-stress nickel-plating bath contains no organic wetting agents that cause the nickel to blister at high temperatures.

  12. 30 CFR 22.10 - Approval plate.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... MINING PRODUCTS PORTABLE METHANE DETECTORS § 22.10 Approval plate. (a) Attachment to be made by manufacturers. (1) Manufacturers shall attach, stamp, or mold an approval plate on each permissible methane... follows: Permissible Methane Detector (or Permissible Methane Indicating Detector) Approval No. ___...

  13. Layered Plating Specimens For Mechanical Tests

    NASA Technical Reports Server (NTRS)

    Thompson, Linda B.; Flowers, Cecil E.

    1991-01-01

    Layered specimens readily made in standard sizes for tensile and other tests of mechanical properties. Standard specimen of metal ordinarily difficult to plate to standard grip thickness or diameter made by augmentation with easier-to-plate material followed by machining to standard size and shape.

  14. 49 CFR 213.123 - Tie plates.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 4 2014-10-01 2014-10-01 false Tie plates. 213.123 Section 213.123 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION TRACK SAFETY STANDARDS Track Structure § 213.123 Tie plates. (a) In Classes 3 through 5...

  15. 49 CFR 213.123 - Tie plates.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Tie plates. 213.123 Section 213.123 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION TRACK SAFETY STANDARDS Track Structure § 213.123 Tie plates. (a) In Classes 3 through 5...

  16. 49 CFR 213.123 - Tie plates.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 4 2011-10-01 2011-10-01 false Tie plates. 213.123 Section 213.123 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION TRACK SAFETY STANDARDS Track Structure § 213.123 Tie plates. (a) In Classes 3 through 5...

  17. 49 CFR 213.123 - Tie plates.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 4 2013-10-01 2013-10-01 false Tie plates. 213.123 Section 213.123 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION TRACK SAFETY STANDARDS Track Structure § 213.123 Tie plates. (a) In Classes 3 through 5...

  18. Cycloid kinematics of relative plate motion

    SciTech Connect

    Cronin, V.S.

    1987-11-01

    The trajectory of a point on one plate as observed from another plate is generally a complex curve and not a small circle around a single axis of relative motion, as is commonly assumed. The shape of the relative-motion path is given the general name spherical cycloid because of its morphological similarity to cycloid planetary trajectories described by early astronomers. The cycloid relative-motion model predicts that the following phenomena occur during finite displacements: (1) the relative velocity and the curvature of the trajectory of a point on one plate relative to another plate varies systematically; (2) plates wobble relative to one another; and (3) the angle of convergence and/or divergence varies systematically along the length of any given transform fault. The small-circle relative-motion model, whereby transform faults have been considered lines of pure slip along which crust is conserved, is not generally valid for finite relative displacements.

  19. Shock wave absorber having apertured plate

    DOEpatents

    Shin, Yong W.; Wiedermann, Arne H.; Ockert, Carl E.

    1985-01-01

    The shock or energy absorber disclosed herein utilizes an apertured plate maintained under the normal level of liquid flowing in a piping system and disposed between the normal liquid flow path and a cavity pressurized with a compressible gas. The degree of openness (or porosity) of the plate is between 0.01 and 0.60. The energy level of a shock wave travelling down the piping system thus is dissipated by some of the liquid being jetted through the apertured plate toward the cavity. The cavity is large compared to the quantity of liquid jetted through the apertured plate, so there is little change in its volume. The porosity of the apertured plate influences the percentage of energy absorbed.

  20. Shock wave absorber having apertured plate

    DOEpatents

    Shin, Y.W.; Wiedermann, A.H.; Ockert, C.E.

    1983-08-26

    The shock or energy absorber disclosed herein utilizes an apertured plate maintained under the normal level of liquid flowing in a piping system and disposed between the normal liquid flow path and a cavity pressurized with a compressible gas. The degree of openness (or porosity) of the plate is between 0.01 and 0.60. The energy level of a shock wave travelling down the piping system thus is dissipated by some of the liquid being jetted through the apertured plate toward the cavity. The cavity is large compared to the quantity of liquid jetted through the apertured plate, so there is little change in its volume. The porosity of the apertured plate influences the percentage of energy absorbed.

  1. Torsion and transverse bending of cantilever plates

    NASA Technical Reports Server (NTRS)

    Reissner, Eric; Stein, Manuel

    1951-01-01

    The problem of combined bending and torsion of cantilever plates of variable thickness, such as might be considered for solid thin high-speed airplane or missile wings, is considered in this paper. The deflections of the plate are assumed to vary linearly across the chord; minimization of the potential energy by means of the calculus of variations then leads to two ordinary linear differential equations for the bending deflections and the twist of the plate. Because the cantilever is analyzed as a plate rather than as a beam, the effect of constraint against axial warping in torsion is inherently included. The application of this method to specific problems involving static deflection, vibration, and buckling of cantilever plates is presented. In the static-deflection problems, taper and sweep are considered.

  2. Optimum design of geodesically stiffened composite plates

    NASA Technical Reports Server (NTRS)

    Guerdal, Zafer; Phillips, John L.

    1988-01-01

    With the goal of tailorability in mind, the in-plane stiffness characteristics of a particular grid stiffened plate configuration under axial and shear loads have been studied. The contribution of the skin to the stiffener network and the resultant skin/rib interaction is analyzed. For the given plate geometry and loads, it is shown that an optimum configuration does exist. To achieve optimally designed practical plate configurations, buckling constraints need to be included in the design. Due to the complex geometry and loading of the plates, a simplified local buckling analysis of isolated stiffeners and triangular skin elements between the stiffeners is considered. Development of a stiffener buckling analysis represent stiffeners as shear deformable plate elements is presented.

  3. Is There Really A North American Plate?

    NASA Astrophysics Data System (ADS)

    Krill, A.

    2011-12-01

    Lithospheric plates are typically identified from earthquake epicenters and evidence such as GPS movements. But no evidence indicates a plate boundary between the North American and South American Plates. Some plate maps show them separated by a transform boundary, but it is only a fracture zone. Other maps show an "undefined plate boundary" or put no boundary between these two plates (check Google images). Early plate maps showed a single large American Plate, quite narrow east of the Caribbean Plate (Le Pichon 1968, Morgan 1968). The North and South American Plates became established by the leading textbook Earth (Press & Siever 1974). On their map, from a Scientific American article by John Dewey (1972), these new plates were separated by an "uncertain plate boundary." The reasons for postulating a North American Plate were probably more psychological than geological. Each of the other continents of the world had its own plate, and North American geologists naturally wanted theirs. Similarly, European geographers used to view Europe as its own continent. A single large plate should again be hypothesized. But the term American Plate would now be ambiguous ("Which plate, North or South?") Perhaps future textbook authors could call it the "Two-American Plate." Textbook authors ultimately decide such global-tectonic matters. I became aware of textbook authors' opinions and influence from my research into the history of Alfred Wegener's continental drift (see Fixists vs. Mobilists by Krill 2011). Leading textbook author Charles Schuchert realized that continental drift would abolish his cherished paleogeographic models of large east-west continents (Eria, Gondwana) and small oceans (Poseiden, Nereis). He and his junior coauthors conspired to keep drift evidence out of their textbooks, from the 1934-editions until the 1969-editions (Physical Geology by Longwell et al. 1969, Historical Geology by Dunbar & Waage 1969). Their textbooks ruled in America. Textbooks

  4. Holographic NDT of thin titanium plate described

    NASA Astrophysics Data System (ADS)

    Yang, R.

    1986-01-01

    Holography is a technique for recording the amplitude and phase changes of an object on a photographic plate in the form of interference fringes. Because the holographic interference method can examine a component in the noncontact mode and is not bothered by complicated part shape or surface roughness, it has been widely used in nondestructive testing. In the rapidly developing aerospace industry, titanium and titanium alloy thin plates are used more and more but local delamination has been found in the production process of bonded titanium plates. Using ultrasonic methods, such defects are difficult to detect and large area delaminations with tightly closed edges can easily be missed. Other testing methods are also unsuccessful. A holographic method was developed for detecting defects in thin titanium plates and several high temperature alloy plates were tested. The testing equipment, methods and results are discussed.

  5. Volcanism in response to plate flexure.

    PubMed

    Hirano, Naoto; Takahashi, Eiichi; Yamamoto, Junji; Abe, Natsue; Ingle, Stephanie P; Kaneoka, Ichiro; Hirata, Takafumi; Kimura, Jun-Ichi; Ishii, Teruaki; Ogawa, Yujiro; Machida, Shiki; Suyehiro, Kiyoshi

    2006-09-01

    Volcanism on Earth is known to occur in three tectonic settings: divergent plate boundaries (such as mid-ocean ridges), convergent plate boundaries (such as island arcs), and hot spots. We report volcanism on the 135 million-year-old Pacific Plate not belonging to any of these categories. Small alkalic volcanoes form from small percent melts and originate in the asthenosphere, as implied by their trace element geochemistry and noble gas isotopic compositions. We propose that these small volcanoes erupt along lithospheric fractures in response to plate flexure during subduction. Minor extents of asthenospheric melting and the volcanoes' tectonic alignment and age progression in the direction opposite to that of plate motion provide evidence for the presence of a small percent melt in the asthenosphere. PMID:16873612

  6. Esophageal culture

    MedlinePlus

    ... for infection-causing germs in a sample of tissue from the esophagus. ... Culture - esophageal ... A sample of tissue from your esophagus is needed. The sample is ... or viruses. Other tests may be done to determine what medicine ...

  7. Endocervical culture

    MedlinePlus

    ... There, they are placed in a special dish (culture). They are then watched to see if bacteria, virus, or fungus grow. Further tests may be done to identify the specific organism and determine the best treatment.

  8. Bile culture

    MedlinePlus

    ... lab. There, it is placed in a special dish called a culture medium to see if bacteria, ... bacteria, virus, or fungus grew in the laboratory dish. Note: Normal value ranges may vary slightly among ...

  9. Esophageal culture

    MedlinePlus

    ... lab. There, it is placed in a special dish (culture) and watched for the growth of bacteria, ... means that no germs grew in the laboratory dish. Normal value ranges may vary slightly among different ...

  10. Blood culture

    MedlinePlus

    Culture - blood ... A blood sample is needed . The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

  11. Gastric culture

    MedlinePlus

    ... test or procedure preparation (3 to 6 years) School age test or procedure preparation (6 to 12 ... immune system. The final results of the gastric culture test may take several weeks. Your provider will ...

  12. Regulation of chondrocyte terminal differentiation in the postembryonic growth plate: the role of the PTHrP-Indian hedgehog axis.

    PubMed

    Farquharson, C; Jefferies, D; Seawright, E; Houston, B

    2001-09-01

    Chondrocyte differentiation during embryonic bone growth is controlled by interactions between PTHrP and Indian hedgehog. We have now determined that the major components of this signaling pathway are present in the postembryonic growth plate. PTHrP was immunolocalized throughout the growth plate, and semiquantitative RT-PCR analysis of maturationally distinct chondrocyte fractions indicated that PTHrP, Indian hedgehog, and the PTH/PTHrP receptor were expressed at similar levels throughout the growth plate. However, patched, the hedgehog receptor, was more highly expressed in proliferating chondrocytes. Although all fractionated cells responded to PTHrP in culture by increasing thymidine incorporation and cAMP production and decreasing alkaline phosphatase activity, the magnitude of response was greatest in the proliferative chondrocytes. Bone morphogenetic proteins are considered likely intermediates in PTHrP signaling. Expression of bone morphogenetic protein-2 and 4--7 was detected within the growth plate, and PTHrP inhibited the expression of bone morphogenetic protein-4 and 6. Although organ culture studies indicated a possible paracrine role for epiphyseal chondrocyte-derived PTHrP in regulating growth plate chondrocyte differentiation, the presence within the postembryonic growth plate of functional components of the PTHrP-Indian hedgehog pathway suggests that local mechanisms intrinsic to the growth plate exist to control the rate of endochondral ossification. PMID:11517192

  13. Tectonic Evolution of the Jurassic Pacific Plate

    NASA Astrophysics Data System (ADS)

    Nakanishi, M.; Ishihara, T.

    2015-12-01

    We present the tectonic evolution of the Jurassic Pacific plate based on magnetic anomly lineations and abyssal hills. The Pacific plate is the largest oceanic plate on Earth. It was born as a microplate aroud the Izanagi-Farallon-Phoenix triple junction about 192 Ma, Early Jurassic [Nakanishi et al., 1992]. The size of the Pacific plate at 190 Ma was nearly half that of the present Easter or Juan Fernandez microplates in the East Pacific Rise [Martinez et at, 1991; Larson et al., 1992]. The plate boundary surrounding the Pacific plate from Early Jurassic to Early Cretaceous involved the four triple junctions among Pacific, Izanagi, Farallon, and Phoenix plates. The major tectonic events as the formation of oceanic plateaus and microplates during the period occurred in the vicinity of the triple junctions [e.g., Nakanishi and Winterer, 1998; Nakanishi et al., 1999], implying that the study of the triple junctions is indispensable for understanding the tectonic evolution of the Pacific plate. Previous studies indicate instability of the configuration of the triple junctions from Late Jurassic to Early Cretaceous (155-125 Ma). On the other hand, the age of the birth of the Pacific plate was determined assuming that all triple junctions had kept their configurations for about 30 m.y. [Nakanishi et al., 1992] because of insufficient information of the tectonic history of the Pacific plate before Late Jurassic.Increase in the bathymetric and geomagnetic data over the past two decades enables us to reveal the tectonic evolution of the Pacific-Izanagi-Farallon triple junction before Late Jurassic. Our detailed identication of magnetic anomaly lineations exposes magnetic bights before anomaly M25. We found the curved abyssal hills originated near the triple junction, which trend is parallel to magnetic anomaly lineations. These results imply that the configuration of the Pacific-Izanagi-Farallon triple junction had been RRR before Late Jurassic.

  14. Parallel Plate System for Collecting Data Used to Determine Viscosity

    NASA Technical Reports Server (NTRS)

    Kaukler, William (Inventor); Ethridge, Edwin C. (Inventor)

    2013-01-01

    A parallel-plate system collects data used to determine viscosity. A first plate is coupled to a translator so that the first plate can be moved along a first direction. A second plate has a pendulum device coupled thereto such that the second plate is suspended above and parallel to the first plate. The pendulum device constrains movement of the second plate to a second direction that is aligned with the first direction and is substantially parallel thereto. A force measuring device is coupled to the second plate for measuring force along the second direction caused by movement of the second plate.

  15. Mass production of spores of lactic acid-producing Rhizopus oryzae NBRC 5384 on agar plate.

    PubMed

    Yamane, Tsuneo; Tanaka, Ryosuke

    2013-01-01

    Mass production of sporangiospores (spores) of Rhizopus oryzae NBRC 5384 (identical to NRRL 395 and ATCC 9363) on potato-dextrose-agar medium was studied aiming at starting its L(+)-lactic acid fermentation directly from spore inoculation. Various parameters including harvest time, sowed spore density, size of agar plate, height of air space, and incubation mode of plate (agar-on-bottom or agar-on-top) were studied. Ordinarily used shallow Petri dishes were found out to be unsuitable for the full growth of R. oryzae sporangiophores. In a very wide range of the sowed spore density, the smaller it was, the greater the number of the harvested spores was. It was also interesting to find out that R. oryzae grown downward vertically with a deep air space in an agar-on-top mode gave larger amount of spores than in an agar-on-bottom mode at 30°C for 7-day cultivation. Scale-up of the agar plate culture from 26.4 to 292 cm(2) was studied, resulting in the proportional relationship between the number of the harvested spores/plate and the plate area in the deep Petri dishes. The number of plates of 50 cm in diameter needed for 100 m(3) industrial submerged fermentation started directly from 2 × 10(5) spores/mL inoculum size was estimated as about 6, from which it was inferred that such a fermentation would be feasible. Designing a 50 cm plate and a method of spreading and collecting the spores were suggested. Bioprocess technological significance of the "full-scale industrial submerged fermentation started directly from spore inoculation omitting pre-culture" has been discussed. PMID:23658025

  16. Urine culture - catheterized specimen

    MedlinePlus

    Culture - urine - catheterized specimen; Urine culture - catheterization; Catheterized urine specimen culture ... urinary tract infections may be found in the culture. This is called a contaminant. You may not ...

  17. Spirit's Tracks around 'Home Plate'

    NASA Technical Reports Server (NTRS)

    2006-01-01

    [figure removed for brevity, see original site] Annotated Version

    This portion of an image acquired by the Mars Reconnaissance Orbiter's High Resolution Imaging Science Experiment camera shows the Spirit rover's winter campaign site. The rover is visible. So is the 'Low Ridge' feature where Spirit was parked with an 11-degree northerly tilt to maximize sunlight on the solar panels during the southern winter season. Tracks made by Spirit on the way to 'Home Plate' and to and from 'Tyrone,' an area of light-toned soils exposed by rover wheel motions, are also evident. The original image is catalogued as PSP_001513_1655_red and was taken Sept. 29, 2006.

    NASA's Jet Propulsion Laboratory, a division of the California Institute of Technology in Pasadena, manages the Mars Reconnaissance Orbiter for NASA's Science Mission Directorate, Washington. Lockheed Martin Space Systems, Denver, is the prime contractor for the project and built the spacecraft. The High Resolution Imaging Science Experiment is operated by the University of Arizona, Tucson, and the instrument was built by Ball Aerospace and Technology Corp., Boulder, Colo.

  18. The jubilee of plate tectonics

    NASA Astrophysics Data System (ADS)

    Glen, William; Frankel, Henry

    What should one expect at a birthday gathering for an idea that suffered disdain from birth, but was later transformed overnight into the ruling global theory of the Earth sciences? Tom Hilde and Richard Carlson (Texas A&M University, College Station, Tex.), under the auspices of the Texas A&M Geodynamics Research Institute, with the cooperation of the National Aeronautics and Space Administration (NASA) , the Inter-Union Commission on the Lithosphere, and the U.S. Geodynamics Committee, convened an unprecedented gathering of the makers of Earth science's modern revolution and a large number of their foremost disciples at Texas A&M April 22- 24 , 1987.Those who missed the show will have to read for weeks to learn of the panoply of profound new developments across the broad front of research bearing on plate tectonics, but those with an interest in their own intellectual genealogy will not be able to recapture the sense of historic moment that pervaded Rudder Tower Conference Center as many of the old revolutionaries opened windows on the past. Of the 20 key participants or originators who were invited to speak, all but four sent abstracts. Pleased and surprised by this strong response, the convenors found themselves in the rather difficult position of having to limit the number of talks by nonoriginators to 20. The unique assembly also provided the first opportunity to video interviews with more than a dozen of the originators.

  19. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus

    PubMed Central

    Cold, Emma R.; Freyria, Nastasia J.; Martínez Martínez, Joaquín; Fernández Robledo, José A.

    2016-01-01

    The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham’s F12–5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham’s F12–5% FBS– 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham’s F12–5% FBS– 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. PMID:27149378

  20. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus.

    PubMed

    Cold, Emma R; Freyria, Nastasia J; Martínez Martínez, Joaquín; Fernández Robledo, José A

    2016-01-01

    The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham's F12-5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham's F12-5% FBS- 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham's F12-5% FBS- 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. PMID:27149378

  1. Tissue and cell studies of the growth plate in the chondrodysplasias.

    PubMed

    Horton, W A; Campbell, D; Machado, M A; Aulthouse, A L; Ahmed, S; Ellard, J T

    1989-09-01

    As the morphologic expression of the chondrocytic differentiation pathway responsible for bone development and growth, the growth plate has been investigated extensively in the chondrodysplasias. Unique morphologic abnormalities identified in many disorders have provided insight into pathogenetic mechanisms and have been useful diagnostically and nosologically. Biochemical studies have detected evidence of type II collagen defects in patients having disorders in the achondrogenesis type II-spondyloepiphyseal dysplasias (SED) congenita family of chondrodysplasias. Most promising may be the cell culture systems now being developed for human chondrocytes. Preliminary results suggest that they will allow the cellular and molecular biology of the dysplastic growth plate to be directly analyzed. PMID:2816990

  2. Lee plate equations for electroded quartz crystal plates with the consideration of electrode density and stiffness.

    PubMed

    Wang, Ji; Chen, Guojun; Du, Jianke

    2008-02-01

    Lee plate equations for high-frequency vibrations of piezoelectric plates have been established and improved over the last decades with the sole objective of obtaining the accurate prediction of frequency and mode shapes to aid crystal resonator design. The latest improvement includes extra terms related to derivatives of the flexural displacement to adjust the accuracy and consider electrodes for practical applications. As part of the efforts to make the equations applicable for resonator design with the improved frequency accuracy and consideration of electrodes, we derived Lee plate equations for electroded plates by changing the integration limits in the dimension reduction procedure to signify the dominant role of the crystal plate. By modifying the density terms in plate equations to include the contribution of both electrode stiffness and density, the accuracy of the thickness-shear vibration frequency of electroded plates is improved for commonly used electrode materials. PMID:18334357

  3. Tissue in Cube: In Vitro 3D Culturing Platform with Hybrid Gel Cubes for Multidirectional Observations.

    PubMed

    Hagiwara, Masaya; Kawahara, Tomohiro; Nobata, Rina

    2016-07-01

    An in vitro 3D culturing platform enabling multidirectional observations of 3D biosamples is presented. The 3D structure of biosamples can be recognized without fluorescence. The cubic platform employs two types of hydrogels that are compatible with conventional culture dishes or well plates, facilitating growth in culture, ease of handling, and viewing at multiple angles. PMID:27128576

  4. Reducing chromium losses from a chromium plating bath. 1987 summer intern report. Project conducted at New Dimension Plating, Hutchinson, Minnesota

    SciTech Connect

    Achman, D.

    1987-12-31

    The company employs about forty people and operates for one or two eight hour shifts with an average of 315 racks of chrome plating per eight hour day. They plate a variety of metals including copper, nickel, gold, brass and chromium. Chromium is the major metal plated and is usually the last step in plating cycle. Most parts are copper plated and then nickel plated in preparation for chrome plating. The main difference between New Dimension Plating and other plating shops is the variety of parts plated. As New Dimension Plating is a job shop, a wide range of parts such as motorcycle accessories, stove parts, and custom items are metal finished. The plating lines are manual, meaning employees dip the racks into the tanks by hand. This fact along with the fact that parts vary greatly in size and shape accounts for the significant drag-out on the chromium plating line.

  5. Anaerobic bag culture method.

    PubMed

    Rosenblatt, J E; Stewart, P R

    1975-06-01

    In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method. PMID:1100671

  6. Anaerobic bag culture method.

    PubMed Central

    Rosenblatt, J E; Stewart, P R

    1975-01-01

    In a new method of anaerobic culture, a transparent, gas-impermeable bag is used and the anaerobic environment is established with copper sulfate-saturated steel wool. An Alka-Seltzer tablet generates carbon dioxide. The agar plate surface can be inspected through the bag at any time without interrupting the anaerobic atmosphere or disturbing other specimens. Methylene blue indicator strips are completely reduced by 4 h after the bag is set up and have remained reduced for as long as 3 weeks. Growth of 16 different stock culture anaerobes was generally equivalent by the bag and GasPak jar methods. Yield and growth of anaerobic isolates also were equivalent with 7 of 10 clinical specimens; from the other 3 specimens, 13 isolates were recovered, 5 by both the bag and jar methods and the rest by one method or the other. No consistent differences were found between the anaerobic bag and GasPak jar methods in the yield of anaerobes from clinical specimens. Early growth (24 h of incubation) of anaerobes from one specimen was detected with the bag method. Images PMID:1100671

  7. Shells on a Sphere: Tectonic Plate Motion and Plate Boundary Deformation

    NASA Astrophysics Data System (ADS)

    Apel, Edwin Victor, III

    Plate motion models have matured from being based only on geology and seismicity to incorporating space-based geodetic methods like GPS. I use a block modeling approach to incorporate both rigid block rotation and near-boundary elastic strain accumulation effects in a formal inversion of GPS velocities. Independent Okhotsk and Amurian microplate motions are tested using GPS velocities that constrain the plate kinematics of northeast Asia. Modeling favors scenarios with independent OKH and AMU motion, based on the application of F-test statistics. The plate-motion parameters of the independent plates are consistent with the kinematics inferred from earthquake focal mechanism solutions along their boundaries. GPS-measured velocities (15 from continuously recording stations within the stable India plate interior) geodetically constrain India plate motion, intraplate strain, and plate boundary deformation around the India plate. Dense station coverage from previously published studies allows rigorous testing of boundary parameterizations. I develop robust India plate motion parameters and see good agreement between predicted plate directions from the preferred model and the seismological data. Available GPS data in and around the Aegean region is combined and used to evaluate plate motion models, elastic plate boundary deformation and its relationship to seismogenic coupling along the Hellenic subduction zone. The A.D. 365 M ˜8.4, the A.D. 1303 M˜8 Crete suggest that portions of the plate interface must be locked. The primary focus of this study is to examine potential upper plate deformation resulting from a locked subduction interface, active hanging-wall extension, or both. I consider multiple model scenarios in an attempt to interpret the both the horizontal and vertical geodetic signals in the region and its implications for earthquake hazard assessment.

  8. Colorimetric microtiter plate receptor-binding assay for the detection of freshwater and marine neurotoxins targeting the nicotinic acetylcholine receptors

    USGS Publications Warehouse

    Rubio, Fernando; Kamp, Lisa; Carpino, Justin; Faltin, Erin; Loftin, Keith A.; Molgó, Jordi; Aráoz, Rómulo

    2014-01-01

    Anatoxin-a and homoanatoxin-a, produced by cyanobacteria, are agonists of nicotinic acetylcholine receptors (nAChRs). Pinnatoxins, spirolides, and gymnodimines, produced by dinoflagellates, are antagonists of nAChRs. In this study we describe the development and validation of a competitive colorimetric, high throughput functional assay based on the mechanism of action of freshwater and marine toxins against nAChRs. Torpedo electrocyte membranes (rich in muscle-type nAChR) were immobilized and stabilized on the surface of 96-well microtiter plates. Biotinylated α-bungarotoxin (the tracer) and streptavidin-horseradish peroxidase (the detector) enabled the detection and quantitation of anatoxin-a in surface waters and cyclic imine toxins in shellfish extracts that were obtained from different locations across the US. The method compares favorably to LC/MS/MS and provides accurate results for anatoxin-a and cyclic imine toxins monitoring. Study of common constituents at the concentrations normally found in drinking and environmental waters, as well as the tolerance to pH, salt, solvents, organic and inorganic compounds did not significantly affect toxin detection. The assay allowed the simultaneous analysis of up to 25 samples within 3.5 h and it is well suited for on-site or laboratory monitoring of low levels of toxins in drinking, surface, and ground water as well as in shellfish extracts.

  9. Design of a titering assay for lentiviral vectors utilizing direct extraction of DNA from transduced cells in microtiter plates

    PubMed Central

    Murphy, Michele E; Vin, Chintan D; Slough, Megan M; Gombotz, Wayne R; Kelley-Clarke, Brenna

    2016-01-01

    Using lentiviral vector products in clinical applications requires an accurate method for measuring transduction titer. For vectors lacking a marker gene, quantitative polymerase chain reaction is used to evaluate the number of vector DNA copies in transduced target cells, from which a transduction titer is calculated. Immune Design previously described an integration-deficient lentiviral vector pseudotyped with a modified Sindbis virus envelope for use in cancer immunotherapy (VP02, of the ZVex platform). Standard protocols for titering integration-competent lentiviral vectors employ commercial spin columns to purify vector DNA from transduced cells, but such columns are not optimized for isolation of extrachromosomal (nonintegrated) DNA. Here, we describe a 96-well transduction titer assay in which DNA extraction is performed in situ in the transduction plate, yielding quantitative recovery of extrachromosomal DNA. Vector titers measured by this method were higher than when commercial spin columns were used for DNA isolation. Evaluation of the method’s specificity, linear range, and precision demonstrate that it is suitable for use as a lot release assay to support clinical trials with VP02. Finally, the method is compatible with titering both integrating and nonintegrating lentiviral vectors, suggesting that it may be used to evaluate the transduction titer for any lentiviral vector. PMID:26942209

  10. Colorimetric microtiter plate receptor-binding assay for the detection of freshwater and marine neurotoxins targeting the nicotinic acetylcholine receptors.

    PubMed

    Rubio, Fernando; Kamp, Lisa; Carpino, Justin; Faltin, Erin; Loftin, Keith; Molgó, Jordi; Aráoz, Rómulo

    2014-12-01

    Anatoxin-a and homoanatoxin-a, produced by cyanobacteria, are agonists of nicotinic acetylcholine receptors (nAChRs). Pinnatoxins, spirolides, and gymnodimines, produced by dinoflagellates, are antagonists of nAChRs. In this study we describe the development and validation of a competitive colorimetric, high throughput functional assay based on the mechanism of action of freshwater and marine toxins against nAChRs. Torpedo electrocyte membranes (rich in muscle-type nAChR) were immobilized and stabilized on the surface of 96-well microtiter plates. Biotinylated α-bungarotoxin (the tracer) and streptavidin-horseradish peroxidase (the detector) enabled the detection and quantitation of anatoxin-a in surface waters and cyclic imine toxins in shellfish extracts that were obtained from different locations across the US. The method compares favorably to LC/MS/MS and provides accurate results for anatoxin-a and cyclic imine toxins monitoring. Study of common constituents at the concentrations normally found in drinking and environmental waters, as well as the tolerance to pH, salt, solvents, organic and inorganic compounds did not significantly affect toxin detection. The assay allowed the simultaneous analysis of up to 25 samples within 3.5 h and it is well suited for on-site or laboratory monitoring of low levels of toxins in drinking, surface, and ground water as well as in shellfish extracts. PMID:25260255

  11. Scaling of energy absorbing composite plates

    NASA Technical Reports Server (NTRS)

    Jackson, Karen; Lavoie, J. Andre; Morton, John

    1994-01-01

    The energy absorption response and crushing characteristics of geometrically scaled graphite-Kevlar epoxy composite plates were investigated. Two different trigger mechanisms including notch, and steeple geometries were incorporated into the plate specimens to initiate crushing. Sustained crushing was achieved with a new test fixture which provided lateral support to prevent global buckling. Values of specific sustained crushing stress (SSCS) were obtained which were lower than values reported for tube specimens from previously published data. Two sizes of hybrid plates were fabricated; a baseline or model plate, and a full-scale plate with inplane dimensions scaled by a factor of two. The thickness dimension of the full-scale plates was increased using two different techniques: the ply-level method in which each ply orientation in the baseline laminate stacking sequence is doubled, and the sublaminate technique in which the baseline laminate stacking sequence is repeated as a group. Results indicated that the SSCS has a small dependence on trigger mechanism geometry. However, a reduction in the SSCS of 10-25% was observed for the full-scale plates as compared with the baseline specimens, indicating a scaling effect in the crushing response.

  12. Scaling of energy absorbing composite plates

    NASA Technical Reports Server (NTRS)

    Jackson, Karen; Morton, John; Traffanstedt, Catherine; Boitnott, Richard

    1992-01-01

    The energy absorption response and crushing characteristics of geometrically scaled graphite-Kevlar epoxy composite plates were investigated. Three different trigger mechanisms including chamfer, notch, and steeple geometries were incorporated into the plate specimens to initiate crushing. Sustained crushing was achieved with a simple test fixture which provided lateral support to prevent global buckling. Values of specific sustained crushing stress (SSCS) were obtained which were comparable to values reported for tube specimens from previously published data. Two sizes of hybrid plates were fabricated; a baseline or model plate, and a full-scale plate with in-plane dimensions scaled by a factor of two. The thickness dimension of the full-scale plates was increased using two different techniques; the ply-level method in which each ply orientation in the baseline laminate stacking sequence is doubled, and the sublaminate technique in which the baseline laminate stacking sequence is repeated as a group. Results indicated that the SSCS is independent of trigger mechanism geometry. However, a reduction in the SSCS of 10-25 percent was observed for the full-scale plates as compared with the baseline specimens, indicating a scaling effect in the crushing response.

  13. Pulsed electromagnetic field affects intrinsic and endoplasmatic reticulum apoptosis induction pathways in MonoMac6 cell line culture.

    PubMed

    Kaszuba-Zwoinska, J; Chorobik, P; Juszczak, K; Zaraska, W; Thor, P J

    2012-10-01

    Current studies were aimed to elucidate influence of pulsed electromagnetic field stimulation on cell viability and apoptosis induction pathways. For the experimental model we have chosen monocytic cell line MonoMac6 and several apoptosis inducers with different mechanism of death induction like puromycin, colchicine, cyclophosphamide, minocycline and hydrogen peroxide. MonoMac6 cell line was grown at density 1x10(5) cells/well in 96-well culture plates. To induce cell death cell cultures were treated with different apoptosis inducers like puromycin, colchicine, cyclophosphamide, minocycline, hydrogen peroxide and at the same time with pulsed electromagnetic field 50 Hz, 45±5 mT (PEMF) for 4 hour per each stimulation, three times, in 24 hours intervals. Afterwards, cells were harvested for flow cytometry analysis of cell viability measured by annexin V-APC labeled and propidium iodide staining. Expression of apoptosis related genes was evaluated by semi quantitative reverse transcription (RT)-PCR assay. NuPAGE Novex Western blot analysis was carried out for apoptosis inducing factor (AIF) abundance in cytosolic and nuclear extracts of MonoMac6 cells. Puromycin, colchicine and minocycline activated cells and simultaneously treated with PEMF have shown out diminished percentage of annexinV positive (AnV+) cells comparing to controls without PEMF stimulation. MonaMac6 cells puromycin/colchicyne and PEMF treated were to a higher extent double stained (AnV+,PI+), which means increased late apoptotic as well as necrotic (PI+) cells, than non-stimulated controls. On the other hand, minocycline activated cells prior to PEMF treatment showed diminished amount of apoptotic and necrotic (annexin V, annexin V and propidium iodide, propidium iodide positive staining) cells. The opposite effect of PEMF on the percentage of annexin V positively stained cells has been achieved after treatment of MonoMac6 culture with cyclophoshamide and hydrogen peroxide. PEMF enhanced early

  14. Determination of glucose using a coupled-enzymatic reaction with new fluoride selective optical sensing polymeric film coated in microtiter plate wells

    PubMed Central

    Abd-Rabboh, Hisham S. M.; Meyerhoff, Mark E.

    2007-01-01

    The determination of glucose in beverages is demonstrated using newly developed fluoride selective optical sensing polymeric film that contains aluminum (III) octaethylporphyrin (Al[OEP]) ionophore and the chromoionophore ETH7075 cast at the bottom of wells of a 96-well polypropylene microtiter plate. The method uses a dual enzymatic reaction involving glucose oxidase enzyme (GOD) and horseradish peroxidase (HRP), along with an organofluoro-substrate (4-fluorophenol) as the source of fluoride ions. The concentration of fluoride ions after enzymatic reaction is directly proportional to the glucose level in the sample. The method has a detection limit of 0.8 mmol L−1, a linear range of 0.9– 40 mmol L−1 and a sensitivity of 0.125 absorbance unit/decade of glucose concentration. Glucose levels in several beverage samples determined using the proposed method correlate well with a reference spectrophotometric enzyme method based on detection of hydrogen peroxide using bromopyrogallol red dye (BPR). The new method can also be used to determine H2O2 concentrations in the 0.1 - 50 mmol L−1 range using a single enzymatic reaction involving H2O2 oxidation of 4-fluorophenol catalyzed by HRP. The methodology could potentially be used to detect a wide range of substrates for which selective oxidase enzymes exist (to generate H2O2), with the high throughput of simple microtiter plate detection scheme. PMID:18480896

  15. Electrochemical Machining of Metal Plates

    SciTech Connect

    Cooper, J F; Evans, M C

    2005-03-04

    Electrochemical machining (ECM) with concentrated sodium chlorate electrolyte was used to rapidly cut a circular groove (13 cm diameter, 0.2 cm wide) through a 0.15 cm thick sheet of steel--thus opening a 5-inch porthole in as little as 10 minutes. The most favorable operating conditions were: T = 22 C; chlorate concentration 600 g NaClO{sub 3}/liter-solution; electric power of 100 A at 10 V; and flow of 0.5 l/s at a pressure drop of 10 kPa (1.5 psi). The porthole may be removed entirely by electrochemical means, or the electrochemical dissolution may continue until only thin membrane remains that is subsequently cut with a utility knife. An array of thermocouples was used to track temperature in the flowing solution and in the trough being machined; the maximum increase in temperature in the trough was 5 C over that of the flowing electrolyte which increased in temperature by 19 C because of power dissipation. ECM is shown feasible for rapid perforation of plates of ferrous and non-ferrous metals using portable equipment and commercial batteries. The technique can be extended to cut perforations of arbitrary shape through non-planar surfaces using a deformable ring cathode. Analysis of the power requirements for electrolyte flow and ECM indicate a total system weight of less than 45 kg (100 lb) using a commercial NiMH battery. The technique is recommended for reduction to practice and demonstration on full scale as an engineering prototype.

  16. Multiplex profiling of cellular invasion in 3D cell culture models.

    PubMed

    Burgstaller, Gerald; Oehrle, Bettina; Koch, Ina; Lindner, Michael; Eickelberg, Oliver

    2013-01-01

    To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states. PMID:23671660

  17. Multiplex Profiling of Cellular Invasion in 3D Cell Culture Models

    PubMed Central

    Burgstaller, Gerald; Oehrle, Bettina; Koch, Ina; Lindner, Michael; Eickelberg, Oliver

    2013-01-01

    To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states. PMID:23671660

  18. Modeling Residual Stresses in Ceramic Plates

    SciTech Connect

    Cantavella, V.; Moreno, A.; Mezquita, A.; Reig, Y.

    2008-02-15

    The generation of residual stresses during cooling of layered ceramic plates has been modeled. Each plate comprises a body and two thin layers (engobe and glaze). The model takes into account two types of stresses: thermal stresses, resulting from temperature gradients inside the plate during cooling, and the stresses produced by the mismatch of the coefficient of thermal expansion (CTE) of the layers. The body has been simulated using a linear viscoelastic constitutive equation. The engobe and the glaze layer have been considered as elastic solids below a certain temperature (setting temperature: T{sub a}). Above T{sub a} these two layers have no mechanical influence on the body.

  19. Bi-coolant flat plate solar collector

    NASA Astrophysics Data System (ADS)

    Chon, W. Y.; Green, L. L.

    The feasibility study of a flat plate solar collector which heats air and water concurrently or separately was carried out. Air flows above the collector absorber plate, while water flows in tubes soldered or brazed beneath the plate. The collector efficiencies computed for the flow of both air and water are compared with those for the flow of a single coolant. The results show that the bi-coolant collector efficiency computed for the entire year in Buffalo, New York is higher than the single-coolant collector efficiency, although the efficiency of the water collector is higher during the warmer months.

  20. Ion plating with an induction heating source

    NASA Technical Reports Server (NTRS)

    Spalvins, T.; Brainard, W. A.

    1976-01-01

    Induction heating is introduced as an evaporation heat source in ion plating. A bare induction coil without shielding can be directly used in the glow discharge region with no arcing. The only requirement is to utilize an rf inductive generator with low operating frequency of 75 kHz. Mechanical simplicity of the ion plating apparatus and ease of operation is a great asset for industrial applications; practically any metal such as nickel, iron, and the high temperature refractories can be evaporated and ion plated.