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Sample records for 96-well culture plates

  1. 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

    2015-05-14

    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.

  2. 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

  3. 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.

  4. An automated 96-well-plate loader for FACScan.

    PubMed

    Illges, H

    1999-01-01

    Staining multiple samples for data acquisition with a flow cytometer is done in 96-well plates to save time and make large data assessment in one working day possible. However, the inability of the FACScan to take up the samples from 96-well plates is a major drawback. In order to avoid the individual transfer of samples to tubes, we have developed a system, which allows using the FACScan with 96-well plates. The machine consists of a programmable control module and a loader which moves the 96-well plate in 3 axis well by well along the sample collector. The machine is equipped with a wash buffer tank to avoid cross contamination of samples and a shaking option to avoid sedimentation of cells during acquisition. The machine can be further developed into a full automatic loader if connected to the FACS Station. In 24 hours about 7,000 samples with 10,000 cells each can be acquired.

  5. Induction of apoptosis by intracellular potassium ion depletion: using the fluorescent dye PBFI in a 96-well plate method in cultured lung cancer cells.

    PubMed

    Andersson, B; Janson, V; Behnam-Motlagh, P; Henriksson, R; Grankvist, K

    2006-09-01

    Depletion of intracellular potassium ions (K+) is necessary for cells to shrink, activate caspases and induce DNA fragmentation, events which are features of apoptosis. Here we describe a 96-well plate method using the cell permeable form of K+ binding benzofuran isophtalate (PBFI-AM) to measure intracellular K+ content in relation to untreated control. Cultured human pulmonary mesothelioma cells (P31) and small-cell lung cancer cells (U1690) were treated with K+ flux modulators in order to deprive the cells of intracellular K+. The combination of K+ influx inhibition with 10 micromol/L bumetanide plus 10 micromol/L ouabain and K+ efflux stimulation with 3 mg/L amphotericin B or 5 micromol/L nigericin efficiently reduced the intracellular K+ content after 3 h. Manipulation of K+ fluxes with subsequent intracellular K+ depletion induced apoptosis of lung cancer cells, as detected by caspase-3 activity after 3 h K+ depletion followed by 24 h proliferation and TUNEL positive staining after 48 h proliferation. We concluded that the PBFI-AM assay was a useful tool to determine intracellular K+ content in relation to untreated control, and that intracellular K+ depletion of lung cancer cells by clinically used drugs of relevant concentrations induced apoptosis. These findings may lead to novel therapeutic strategies in the treatment of lung cancer.

  6. 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...

  7. Performance evaluation of 3D polystyrene 96-well plates with human neural stem cells in a calcium assay.

    PubMed

    Lai, Yinzhi; Kisaalita, William S

    2012-08-01

    In this study, we have generated a high-throughput screening (HTS)-compatible 3D cell culture platform by chemically "welding" polystyrene scaffolds into standard 2D polystyrene 96-well plates. The variability of scaffolds was minimized by introducing automation into the fabrication process. The fabricated 3D cell culture plates were compared with several commercially available 3D cell culture platforms with light and scanning electron microscopy. Voltage-gated calcium channel functionality was used to access the Z' factors of all plates, including a 2D standard plate control. It was found that with the No-Wash Fluo-4 calcium assay and neural progenitor cells, all plates display acceptable Z' factors for use in HTS. The plates with "welded" polystyrene scaffolds have several advantages, such as being versatile and economical, and are ready to use off the shelf. These characteristics are especially desired in HTS preclinical drug discovery applications.

  8. 96-Well plate assays for measuring collagenase activity using (3)H-acetylated collagen.

    PubMed

    Koshy, P J; Rowan, A D; Life, P F; Cawston, T E

    1999-11-15

    We describe two alternative assays for measuring collagenolytic activity using (3)H-acetylated collagen. Both assays have been developed for the 96-well plate format and measure the amount of radiolabeled collagen fragments released into the supernatant from an insoluble (3)H-acetylated collagen fibril preparation. The first method separates digested solubilized fragments from the intact fibril by sedimentation of the undigested collagen by centrifugation. The second method achieves this separation by filtration of the supernatant through the membrane of a 96-well filtration plate which retains the undigested collagen fibril. Both methods give linear dose- and time-dependent responses of collagenase activity > or = 70% of total collagen lysis. In addition, both assays can be simply modified to measure tissue inhibitors of metalloproteinases (TIMPs) inhibitory activity, which is also linear between 20 and 75% of total collagen lysis with the amount of TIMP added.

  9. A 96-well plate assay for CYP4503A induction using cryopreserved human hepatocytes.

    PubMed

    Kamiguchi, Naomi; Aoyama, Eiji; Okuda, Teruaki; Moriwaki, Toshiya

    2010-11-01

    A reliable and practical CYP3A induction assay with cryopreserved human hepatocytes in a 96-well format was developed. Various 96-well plates with different basement membrane were evaluated using prototypical inducers, rifampicin, phenytoin, and carbamazepine. Thin-layer (TL) Matrigel was found to yield the highest basal and induced levels of CYP3A activity as determined by testosterone 6β-hydroxylation. Concentration-dependent CYP3A induction of rifampicin was reproducible with the EC(50) values of 0.36 ± 0.28 μM from four batches of human hepatocytes using the 96-well plate with TL Matrigel. The rank order of induction potency for nine inducers or noninducers at a concentration of 10 μM were well comparable among the multiple donors, by expressing the results as percentage of change compared with the positive control, 10 μM rifampicin. Cotreatment of avasimibe or efavirenz with 10 μM rifampicin was found to reduce CYP3A activities induced by rifampicin at a lower rate than treatment with rifampicin alone, whereas treatment with phenobarbital and carbamazepine had no effect. From a comparison of induced CYP3A activities and gene expression levels, there were compounds that would cause induction of CYP3A4 mRNA but not activity, presumably due to their inhibitory effect on CYP3A activity. The cotreatment assay of test compound with rifampicin allows us to exclude the false-negative results caused by the cytotoxicity and/or the mechanism-based inactivation, when the drug candidate's ability for CYP3A induction is evaluating the enzyme activity. This 96-well plate assay, which is robust, reproducible, and convenient, has demonstrated the paramount applicability to the early drug discovery stage.

  10. Streptomycetes in micro-cultures: growth, production of secondary metabolites, and storage and retrieval in the 96-well format.

    PubMed

    Minas, W; Bailey, J E; Duetz, W

    2000-12-01

    Mycelium-forming Streptomyces strains were grown in one milliliter liquid micro-cultures in square deep-well microtiter plates. Growth was evaluated with respect to biomass formation and production of secondary metabolites which were found to be very similar in the micro-cultures, bioreactor, and shake flask cultivations, respectively. Despite repetitive sampling and extensive growth on the walls of the wells, no cross contamination occurred. Furthermore, we successfully employed cold storage at -20 degrees C of spore suspensions (in the 96-well format), directly prepared from cultures grown on agar in the microtitre plate. Cultures were retrieved by replicating aliquots from the frozen spore suspensions.

  11. 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

    2014-06-03

    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.

  12. Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics

    PubMed Central

    2015-01-01

    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

  13. A method for high throughput determination of viable bacteria cell counts in 96-well plates

    PubMed Central

    2012-01-01

    Background There are several methods for quantitating bacterial cells, each with advantages and disadvantages. The most common method is bacterial plating, which has the advantage of allowing live cell assessment through colony forming unit (CFU) counts but is not well suited for high throughput screening (HTS). On the other hand, spectrophotometry is adaptable to HTS applications but does not differentiate between dead and living bacteria and has low sensitivity. Results Here, we report a bacterial cell counting method termed Start Growth Time (SGT) that allows rapid and serial quantification of the absolute or relative number of live cells in a bacterial culture in a high throughput manner. We combined the methodology of quantitative polymerase chain reaction (qPCR) calculations with a previously described qualitative method of bacterial growth determination to develop an improved quantitative method. We show that SGT detects only live bacteria and is sensitive enough to differentiate between 40 and 400 cells/mL. SGT is based on the re-growth time required by a growing cell culture to reach a threshold, and the notion that this time is proportional to the number of cells in the initial inoculum. We show several applications of SGT, including assessment of antibiotic effects on cell viability and determination of an antibiotic tolerant subpopulation fraction within a cell population. SGT results do not differ significantly from results obtained by CFU counts. Conclusion SGT is a relatively quick, highly sensitive, reproducible and non-laborious method that can be used in HTS settings to longitudinally assess live cells in bacterial cell cultures. PMID:23148795

  14. Nitrotyrosine Density of Rabbit Urinary Bladder Muscle and Mucosa Measured via Western Blotting and 96-Well Plate Analysis.

    PubMed

    Fitzpatrick, Brittany; Schuler, Catherine; Leggett, Robert E; Levin, Robert M

    2012-01-01

    Purpose. Nitrotyrosine was quantitated in rabbit bladder muscle and mucosa using two analytical systems: Western blotting analyses and a 96-well plate quantitative analysis kit. Materials and Methods. Rabbit bladder muscle and mucosa were obtained from control rabbits. For the Western analysis, the samples were loaded into a SDS page gel and then transferred to a PVDF membrane. The optical density was measured using a Kodak Scanner. Using the 96-well plate, the samples and standards were loaded, incubated with primary and secondary antibody, washed and vacuumed with 10x wash buffer three times between each incubation period. Stop buffer was added to the plate and the results were quantified via the plate reader. Results. For both muscle and mucosa tissue, the optical density readings were linear with tissue concentration; the concentration of nitrotyrosine in the mucosa was significantly higher than in the muscle. However, whereas the Western blot analysis is based on relative optical densities, the 96-well plate kit provides a truly quantitative analysis. Discussion. Mucosa tissue displayed a higher density of nitrotyrosine than did detrusor muscle tissue. This may well be due to the significantly higher metabolic activity of the mucosa compared to the muscle.

  15. 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

    2011-11-09

    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

  16. A 96-well screen filter plate for high-throughput biological sample preparation and LC-MS/MS analysis.

    PubMed

    Peng, Sean X; Cousineau, Martin; Juzwin, Stephen J; Ritchie, David M

    2006-01-01

    A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates.

  17. Thermodynamic equilibrium solubility measurements in simulated fluids by 96-well plate method in early drug discovery.

    PubMed

    Bharate, Sonali S; Vishwakarma, Ram A

    2015-04-01

    An early prediction of solubility in physiological media (PBS, SGF and SIF) is useful to predict qualitatively bioavailability and absorption of lead candidates. Despite of the availability of multiple solubility estimation methods, none of the reported method involves simplified fixed protocol for diverse set of compounds. Therefore, a simple and medium-throughput solubility estimation protocol is highly desirable during lead optimization stage. The present work introduces a rapid method for assessment of thermodynamic equilibrium solubility of compounds in aqueous media using 96-well microplate. The developed protocol is straightforward to set up and takes advantage of the sensitivity of UV spectroscopy. The compound, in stock solution in methanol, is introduced in microgram quantities into microplate wells followed by drying at an ambient temperature. Microplates were shaken upon addition of test media and the supernatant was analyzed by UV method. A plot of absorbance versus concentration of a sample provides saturation point, which is thermodynamic equilibrium solubility of a sample. The established protocol was validated using a large panel of commercially available drugs and with conventional miniaturized shake flask method (r(2)>0.84). Additionally, the statistically significant QSPR models were established using experimental solubility values of 52 compounds.

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

    PubMed Central

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

    2017-01-01

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

  19. 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.

  20. 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.

  1. Mass Spectrometric N-Glycan Analysis of Haptoglobin from Patient Serum Samples Using a 96-Well Plate Format.

    PubMed

    Zhu, Jianhui; Wu, Jing; Yin, Haidi; Marrero, Jorge; Lubman, David M

    2015-11-06

    Alterations in glycosylation of serum glycoproteins can provide unique and highly specific fingerprints of malignancy. Our previous mass spectrometric study revealed that the bifucosylation level of serum haptoglobin was distinctly increased in hepatocellular carcinoma (HCC) patients versus liver cirrhosis of all three major etiologies. We have thus developed a method for the analysis of large numbers of serum samples based on a 96-well plate platform for the evaluation of fucosylation changes of serum haptoglobin between HCC versus cirrhosis. Haptoglobin was isolated from the serum of individual patient samples based on an HPLC column immobilized with antihaptoglobin antibody via hydrazide immobilization chemistry. Only 10 μL of serum was required for glycan extraction and processing for MALDI-QIT mass spectrometry analysis using the 96-well plate format. The bifucosylation degrees of haptoglobin in individuals were calculated using a quantitative glycomics method. The MS data confirmed that the bifucosylated tetra-anntenary glycan was upregulated in HCC samples of all etiologies. This study provides a parallel method for processing glycan content for haptoglobin and evaluating detailed changes in glycan structures for a potentially large cohort of clinical serum samples.

  2. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    PubMed Central

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-01-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format. PMID:27886235

  3. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates

    NASA Astrophysics Data System (ADS)

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-01

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  4. Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

    PubMed

    Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

    2016-11-25

    Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

  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. Development of cystic embryoid bodies with visceral yolk-sac-like structures from mouse embryonic stem cells using low-adherence 96-well plate.

    PubMed

    Yasuda, Emiko; Seki, Yuji; Higuchi, Takatoshi; Nakashima, Fumio; Noda, Tomozumi; Kurosawa, Hiroshi

    2009-04-01

    Cystic embryoid bodies with visceral yolk-sac-like structure (cystic EB-Vs) are used as a model for the study of early extraembryonic tissue formation containing visceral endoderm-like derivatives. In this study, we optimized the cell density of embryonic stem (ES) cells for developing cystic EB-Vs in a low-adherence 96-well plate. When ES cells were seeded at a density of 4000 cells/well, the cystic EB-Vs were most efficiently developed from ES cells via forming multicellular spherical aggregates called embryoid bodies (EBs). The suspension culture in the low-adherence plate was preferable for developing EBs into cystic EB-Vs rather than the attachment culture in the plate coated with 0.1% gelatin. The seeding cell density of 4000 cells/well was always superior to 1000 cells/well in the efficiency of cystic EB-V development. Because the high-cell density culture generally raises the limitation of oxygen and nutrient supplies, we investigated the effects of low-oxygen and low-nutrient conditions on the development of cystic EB-Vs. It was found that low oxygen tension was not a factor for promoting the development of cystic EB-Vs. It was suggested that a low-nutrient medium is preferred for developing cystic EB-Vs rather than a sufficient-nutrient medium. In conclusion, the suspension culture in the low-adherence 96-well plate seeded with 4000 ES cells/well was optimum for developing cystic EB-Vs. The low-nutrient condition may be one of the factors for promoting the development of cystic EB-Vs.

  7. Equilibrium drug solubility measurements in 96-well plates reveal similar drug solubilities in phosphate buffer pH 6.8 and human intestinal fluid.

    PubMed

    Heikkilä, Tiina; Karjalainen, Milja; Ojala, Krista; Partola, Kirsi; Lammert, Frank; Augustijns, Patrick; Urtti, Arto; Yliperttula, Marjo; Peltonen, Leena; Hirvonen, Jouni

    2011-02-28

    This study was conducted to develop a high throughput screening (HTS) method for the assessment of equilibrium solubility of drugs. Solid-state compounds were precipitated from methanol in 96-well plates, in order to eliminate the effect of co-solvent. Solubility of twenty model drugs was analyzed in water and aqueous solutions (pH 1.2 and 6.8) in 96-well plates and in shake-flasks (UV detection). The results obtained with the 96-well plate method correlated well (R(2)=0.93) between the shake-flask and 96-well plates over the wide concentration scale of 0.002-169.2mg/ml. Thereafter, the solubility tests in 96-well plates were performed using fasted state human intestinal fluid (HIF) from duodenum of healthy volunteers. The values of solubility were similar in phosphate buffer solution (pH 6.8) and HIF over the solubility range of 10(2)-10(5)μg/ml. The new 96-well plate method is useful for the screening of equilibrium drug solubility during the drug discovery process and it also allows the use of human intestinal fluid in solubility screening.

  8. 96-well plate-to-plate gravity fluorous solid-phase extraction (F-SPE) for solution-phase library purification.

    PubMed

    Zhang, Wei; Lu, Yimin

    2007-01-01

    Large particle size (125_210 microm) fluorous silica gel bonded with a -SiCH2CH2C8F17 stationary phase has been employed for gravity-driven fluorous solid-phase extraction (F-SPE) on two types of 96-well plates. A 1 or 0.75 g portion of fluorous silica is packed to each well of the 3.5-mL Ex-Blok and the 2.2-mL deep-well filtration plates, respectively. Up to 50 mg of reaction mixture is loaded and then eluted with a fluorophobic solvent (DMSO, DMF, or 85:15 DMF-H2O). Products collected in 96-well receiving plates are directly concentrated on a GeneVac vacuum centrifuge. This simple and highly efficient plate-to-plate F-SPE technique has been demonstrated in the purification of four 96-compound libraries produced by scavenging reactions with 1-(perfluoroctyl)propyl isatoic anhydride (F-IA), amide coupling reactions with 2-chloro-4,6-bis[(perfluorooctyl)propyloxy]-1,3,5-triazine (F-CDMT) or 2,4-dichloro-6-(perfluorooctyl)propyloxy-1,3,5-triazine (F-DCT), and Mitsunobu reactions with fluorous diethyl azodicarboxylate (F-DEAD) and triphenylphosphine (F-TPP). Approximately 80% of products in each library have greater than 85% purity after F-SPE without conducting chromatography.

  9. 96-Well Polycarbonate-Based Microfluidic Titer Plate for High-Throughput Purification of DNA and RNA

    PubMed Central

    Witek, Małgorzata A.; Hupert, Mateusz L.; Park, Daniel S.-W.; Fears, Kirby; Murphy, Michael C.; Soper, Steven A.

    2008-01-01

    We report a simple and effective method for the high-throughput purification of a variety of nucleic acids (NAs) from whole cell lysates or whole blood using a photactivated polycarbonate solid-phase reversible immobilization (PPC-SPRI) microfluidic chip. High-throughput operation was achieved by placing 96 purification beds, each containing an array of 3800 20 µm diameter posts, on a single 3″ × 5″ polycarbonate (PC) wafer fabricated by hot embossing. All beds were interconnected through a common microfluidic network that permitted parallel access through the use of a vacuum and syringe pump for delivery of immobilization buffer (IB) and effluent. The PPC-SPRI purification was accomplished by condensation of NAs onto a UV-modified PC surface in the presence of the IB comprised of polyethylene glycol, NaCl, and ethanol with a composition dependent on the length of the NAs to be isolated and the identity of the sample matrix. The performance of the device was validated by quantification of the recovered material following PCR (for DNA) or RT-PCR (for RNA). The extraction bed load capacity of NAs was 206 ± 93 ng for gDNA and 165 ± 81 ng for TRNA from Escherichia coli. Plate-to-plate variability was found to be 35 ± 10%. The purification process was fast (>30 min) and easy to automate, and the low cost of wafer fabrication makes it appropriate for single-use applications. PMID:18355091

  10. A new approach to the application of solid phase extraction disks with LC-MS/MS for the analysis of drugs on a 96-well plate format.

    PubMed

    Cudjoe, Erasmus; Pawliszyn, Janusz

    2009-11-01

    A new 96-well disk solid phase extraction sample preparation technique which does not involve vacuum pumps integrated with liquid chromatographic tandem mass spectrometric (LC-MS/MS) was developed for high throughput determination of benzodiazepines (nordiazepam, diazepam, lorazepam and oxazepam). In addition, the method completely allows the re-use of the SPE disk membranes for subsequent analyses after re-conditioning. The method utilizes a robotic autosampler for parallel extractions in a 96-well plate format. Results have been presented for independent extractions from three matrices; phosphate buffer solution, urine, and plasma. Factors affecting data reproducibility, extraction kinetics, sample throughput, and reliability of the system were investigated and optimized. A total time required per sample was 0.94 min using 96-well format. Method reproducibility was < or =9% relative standard deviation for all three matrices. Limits of detection and quantitation recorded were respectively in the range 0.02-0.15 and 0.2-2.0 ng/mL with linearity ranging from 0.2 to 500 ng/mL for all matrices.

  11. High-throughput 96-well solvent mediated sonic blending synthesis and on-plate solid/solution stability characterization of pharmaceutical cocrystals.

    PubMed

    Luu, Van; Jona, Janan; Stanton, Mary K; Peterson, Matthew L; Morrison, Henry G; Nagapudi, Karthik; Tan, Helming

    2013-01-30

    A 96-well high-throughput cocrystal screening workflow has been developed consisting of solvent-mediated sonic blending synthesis and on-plate solid/solution stability characterization by XRPD. A strategy of cocrystallization screening in selected blend solvents including water mixtures is proposed to not only manipulate solubility of the cocrystal components but also differentiate physical stability of the cocrystal products. Caffeine-oxalic acid and theophylline-oxalic acid cocrystals were prepared and evaluated in relation to saturation levels of the cocrystal components and stability of the cocrystal products in anhydrous and hydrous solvents. AMG 517 was screened with a number of coformers, and solid/solution stability of the resulting cocrystals on the 96-well plate was investigated. A stability trend was observed and confirmed that cocrystals comprised of lower aqueous solubility coformers tended to be more stable in water. Furthermore, cocrystals which could be isolated under hydrous solvent blending condition exhibited superior physical stability to those which could only be obtained under anhydrous condition. This integrated HTS workflow provides an efficient route in an API-sparing approach to screen and identify cocrystal candidates with proper solubility and solid/solution stability properties.

  12. A high-throughput approach for the determination of pesticide residues in cucumber samples using solid-phase microextraction on 96-well plate.

    PubMed

    Bagheri, Habib; Es'haghi, Ali; Es-haghi, Ali; Mesbahi, Noushin

    2012-08-31

    A high-throughput solid-phase microextraction (SPME) on 96-well plate together with gas chromatography-mass spectrometry (GC-MS) was developed for the determination of some selected pesticides in cucumber samples. Pieces with the length of 1.0 cm of silicon tubing were precisely prepared and then coated on the end part of stainless steel wires. The prepared fibers were positioned in a home-made polytetrafluoroethylene (PTFE)-based constructed ninety-six holes block to have the possibility of simultaneous immersion of the SPME fibers into the center of individual wells. Pesticides such as diazinon, penconazol, tebuconazol, bitertanol, malathion, phosalone and chlorpyrifos-methyl were selected for their highly application in cucumber field. The performances of the SPME fibers, such as intra and inter-fibers reproducibility, were evaluated and the results showed a good similarity in extraction yields. A volume of 1 mL of the aquatic supernatant of the cucumber samples was transferred into the 96-well plate and the array of SPME fibers was applied for the extraction of the selected pesticides. The important parameters influencing the whole extraction process including, organic solvent percent, salt addition, dilution factor, stirring rate and extraction time were optimized. The inter- and intra-day RSD% were found to be less than 15.4%. Limits of detection (LOD) and limits of quantification (LOQ) were below 60 and 180 μg kg(-1), respectively. The coefficient of determination was satisfactory (r(2)>0.99) for all the studied analytes. The developed method was successfully applied to the monitoring of several samples gathered from local markets.

  13. High-throughput determination of fudosteine in human plasma by liquid chromatography-tandem mass spectrometry, following protein precipitation in the 96-well plate format.

    PubMed

    Wen, Jun; Wu, Yiwen; Zhang, Linli; Qi, Yunpeng; Fan, Guorong; Wu, Yutian; Li, Zhen

    2008-05-01

    A 96-well protein precipitation, liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and fully validated for the determination of fudosteine in human plasma. After protein precipitation of the plasma samples (50 microL) by the methanol (150 microL) containing the internal standard (IS), erdosteine, the 96-well plate was vortexed for 5 min and centrifuged for 15 min. The 100 microL supernatant and 100 microL mobile phase were added to another plate and mixed and then the mixture was directly injected into the LC-MS/MS system in the negative ionization mode. The separation was performed on a XB-CN column for 3.0 min per sample using an eluent of methanol-water (60:40, v/v) containing 0.005% formic acid. Multiple reaction monitoring (MRM) using the precursor-product ion transitions m/z 178-->91 and m/z 284-->91 was performed to quantify fudosteine and erdosteine, respectively. The method was sensitive with a lower limit of quantification (LLOQ) of 0.02 microg mL(-1), with good linearity (r>0.999) over the linear range of 0.02-10 microg mL(-1). The within- and between-run precision was less than 5.5% and accuracy ranged from 94.2 to 106.7% for quality control (QC) samples at three concentrations of 0.05, 1 and 8 microg mL(-1). The method was employed in the clinical pharmacokinetic study of fudosteine formulation product after oral administration to healthy volunteers.

  14. A Quantitative High-Throughput 96-well plate Fluorescence Assay for Mechanism-Based Inactivators of Cytochromes P450 Exemplified using CYP2B6

    PubMed Central

    Kenaan, Cesar; Zhang, Haoming; Hollenberg, Paul F.

    2010-01-01

    Mechanism-based inactivators such as bergamottin are useful chemical tools for identifying the roles of specific active-site amino acid residues in the reactions catalyzed by the cytochromes P450 (CYPs or P450s) that are responsible for the metabolism of a wide variety of drugs and endogenous substrates. In clinical settings mechanism-based inactivation of P450s involved in xenobiotic metabolism has the potential to lead to adverse drug-drug interactions and assays to identify and characterize drug candidates as P450 inactivators are important in drug discovery and development. Here we present a quantitative high-throughput protocol for investigating cytochrome P450 mechanism-based inactivators using the example of CYP2B6 and bergamottin to illustrate the finer points of this protocol. This protocol details the adaptation of a 7-ethoxytrifluoromethyl coumarin (7-EFC) O-deethylation fluorescence activity assay to a 96-well microtiter plate format and uses a plate-reader to detect the fluorescence of the product. Compared to previous methods, this protocol requires less P450 and takes significantly less time while greatly increasing throughput. The protocol as written takes approximately two hours to complete. The principles and procedures outlined in this protocol can be easily adapted to other inactivators, P450 isoforms, substrates and plate-readers. PMID:20885377

  15. 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.

  16. Evaluation of drug load and polymer by using a 96-well plate vacuum dry system for amorphous solid dispersion drug delivery.

    PubMed

    Chiang, Po-Chang; Ran, Yingqing; Chou, Kang-Jye; Cui, Yong; Sambrone, Amy; Chan, Connie; Hart, Ryan

    2012-06-01

    It is well recognized that poor dissolution rate and solubility of drug candidates are key limiting factors for oral bioavailability. While numerous technologies have been developed to enhance solubility of the drug candidates, poor water solubility continuously remains a challenge for drug delivery. Among those technologies, amorphous solid dispersions (SD) have been successfully employed to enhance both dissolution rate and solubility of poorly water-soluble drugs. This research reports a high-throughput screening technology developed by utilizing a 96-well plate system to identify optimal drug load and polymer using a solvent casting approach. A minimal amount of drug was required to evaluate optimal drug load in three different polymers with respect to solubility improvement and solid-state stability of the amorphous drug-polymer system. Validation of this method was demonstrated with three marketed drugs as well as with one internal compound. Scale up of the internal compound SD by spray drying further confirmed the validity of this method, and its quality was comparable to a larger scale process. Here, we demonstrate that our system is highly efficient, cost-effective, and robust to evaluate the feasibility of spray drying technology to produce amorphous solid dispersions.

  17. Quantitative analysis of simvastatin and its beta-hydroxy acid in human plasma using automated liquid-liquid extraction based on 96-well plate format and liquid chromatography-tandem mass spectrometry.

    PubMed

    Zhang, Nanyan; Yang, Amy; Rogers, John Douglas; Zhao, Jamie J

    2004-01-27

    An assay based on automated liquid-liquid extraction (LLE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been developed and validated for the quantitative analysis of simvastatin (SV) and its beta-hydroxy acid (SVA) in human plasma. A Packard MultiProbe II workstation was used to convert human plasma samples collected following administration of simvastatin and quality control (QC) samples from individual tubes into 96-well plate format. The workstation was also used to prepare calibration standards and spike internal standards. A Tomtec Quadra 96-channel liquid handling workstation was used to perform LLE based on 96-well plates including adding solvents, separating organic from aqueous layer and reconstitution. SV and SVA were separated through a Kromasil C18 column (50 mm x 2 mm i.d., 5 microm) and detected by tandem mass spectrometry with a TurboIonspray interface. Stable isotope-labeled SV and SVA, 13CD(3)-SV and 13 CD(3)-SVA, were used as the internal standards for SV and SVA, respectively. The automated procedures reduced the overall analytical time (96 samples) to 1/3 of that of manual LLE. Most importantly, an analyst spent only a fraction of time on the 96-well LLE. A limit of quantitation of 50 pg/ml was achieved for both SV and SVA. The interconversion between SV and SVA during the 96-well LLE was found to be negligible. The assay showed very good reproducibility, with intra- and inter-assay precision (%R.S.D.) of less than 7.5%, and accuracy of 98.7-102.3% of nominal values for both analytes. By using this method, sample throughput should be enhanced at least three-fold compared to that of the manual procedure.

  18. Comparison of turbulent-flow chromatography with automated solid-phase extraction in 96-well plates and liquid-liquid extraction used as plasma sample preparation techniques for liquid chromatography-tandem mass spectrometry.

    PubMed

    Zimmer, D; Pickard, V; Czembor, W; Müller, C

    1999-08-27

    Turbulent flow chromatography (TFC) combined with the high selectivity and sensitivity of tandem mass spectrometry (MS-MS) is a new technique for the fast direct analysis of drugs from crude plasma. TFC in the 96-well plate format reduces significantly the time required for sample clean-up in the laboratory. For example, for 100 samples the workload for a technician is reduced from about 8 h by a manual liquid-liquid extraction (LLE) assay to about 1 h in the case of TFC. Sample clean-up and analysis are performed on-line on the same column. Similar chromatographic performance and validation results were achieved using HTLC Turbo-C18 columns (Cohesive Technologies) and Oasis HLB extraction columns (Waters). One 96-well plate with 96 plasma samples is analyzed within 5.25 h, corresponding to 3.3 min per sample. Compared to this LLE and analysis of 96 samples takes about 16 h. Two structurally different and highly protein bound compounds, drug A and drug B, were analyzed under identical TFC conditions and the assays were fully validated for the application to toxicokinetics studies (compliant with Good Laboratory Practices-GLP). The limit of quantitation was 1.00 microg/l and the linear working range covered three orders of magnitude for both drugs. In the case of drug A the quality of analysis by TFC was similar to the reference LLE assay and slightly better than automated solid-phase extraction in 96-well plates. The accuracy was -3.1 to 6.7% and the precision was 3.1 to 6.8% in the case of drug A determined for dog plasma by TFC-MS-MS. For drug B the accuracy was -3.7 to 3.5% and the precision was 1.6 to 5.4% for rat plasma, which is even slightly better than what was achieved with the validated protein precipitation assay.

  19. High-throughput sample preparation for the quantitation of acephate, methamidophos, omethoate, dimethoate, ethylenethiourea, and propylenethiourea in human urine using 96-well-plate automated extraction and high-performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Jayatilaka, Nayana K; Angela Montesano, M; Whitehead, Ralph D; Schloth, Sara J; Needham, Larry L; Barr, Dana Boyd

    2011-07-01

    Acephate, methamidophos, o-methoate, and dimethoate are organophosphorus pesticides, and ethylenethiouria and propylenethiourea are two metabolites from the bisdithiocarbamate fungicide family. They are some of the most widely used pesticides and fungicides in agriculture both domestically and abroad. The existing high-performance liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) method for the measurement of these compounds in human urine was improved by using a 96-well plate format sample preparation; the use of HPLC-MS/MS was comparable with a concentration range of 0.125 to 50 ng/ml. Deuterium-labeled acephate, ethylenethiouria, and methamidophos were used as internal standards. The sample preparation procedure, in the 96-well format with a 0.8-ml urine sample size, uses lyophilization of samples, followed by extraction with dichloromethane. The analytes were chromatographed on a Zorbax SB-C3 (4.6 × 150 mm, 5.0-μm) column with gradient elution by using 0.1% formic acid in aqueous solution (solvent A) and 0.1% formic acid in methanol (solvent B) mobile phase at a flow rate of 1 ml/min. Quantitative analysis was performed by atmospheric pressure chemical ionization source in positive ion mode using multiple-reaction monitoring of the precursor-to-product ion pairs for the analytes on a TSQ Quantum Ultra HPLC-MS/MS. Repeated analyses of urine samples spiked with high (15 ng/ml), medium (5 ng/ml), and low (1 ng/ml) concentrations of the analytes gave relative SDs of <13%. The limits of detection were in the range of 0.004-0.01 ng/ml. The method also has high accuracy, high precision, and excellent extraction recovery. Furthermore, the improved sample preparation method decreased the cost and labor required while effectively doubling the analytic throughput with minimal matrix effect.

  20. Automated liquid-liquid extraction based on 96-well plate format in conjunction with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for the quantitation of methoxsalen in human plasma.

    PubMed

    Yadav, Manish; Contractor, Pritesh; Upadhyay, Vivek; Gupta, Ajay; Guttikar, Swati; Singhal, Puran; Goswami, Sailendra; Shrivastav, Pranav S

    2008-09-01

    A sensitive, specific and high throughput bioanalytical method using automated sample processing via 96-well plate liquid-liquid extraction and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) has been developed for the determination of methoxsalen in human plasma. Plasma samples with ketoconazole as internal standard (IS) were prepared by employing 0.2 mL human plasma in ethyl acetate:dichloromethane (80:20, v/v). The chromatographic separation was achieved on a Waters Acquity UPLC BEH C18 column using isocratic mobile phase, consisting of 10 mM ammonium formate and acetonitrile (60:40, v/v), at a flow rate of 0.5 mL/min. The linear dynamic range was established over the concentration range 1.1-213.1 ng/mL for methoxsalen. The method was rugged and rapid with a total run time of 1.5 min. It was successfully applied to a pivotal bioequivalence study in 12 healthy human subjects after oral administration of 10 mg extended release methoxsalen formulation under fasting condition.

  1. 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.

  2. Stretch Injury of Human Induced Pluripotent Stem Cell Derived Neurons in a 96 Well Format

    PubMed Central

    Sherman, Sydney A.; Phillips, Jack K.; Costa, J. Tighe; Cho, Frances S.; Oungoulian, Sevan R.; Finan, John D.

    2016-01-01

    Traumatic brain injury (TBI) is a major cause of mortality and morbidity with limited therapeutic options. Traumatic axonal injury (TAI) is an important component of TBI pathology. It is difficult to reproduce TAI in animal models of closed head injury, but in vitro stretch injury models reproduce clinical TAI pathology. Existing in vitro models employ primary rodent neurons or human cancer cell line cells in low throughput formats. This in vitro neuronal stretch injury model employs human induced pluripotent stem cell-derived neurons (hiPSCNs) in a 96 well format. Silicone membranes were attached to 96 well plate tops to create stretchable, culture substrates. A custom-built device was designed and validated to apply repeatable, biofidelic strains and strain rates to these plates. A high content approach was used to measure injury in a hypothesis-free manner. These measurements are shown to provide a sensitive, dose-dependent, multi-modal description of the response to mechanical insult. hiPSCNs transition from healthy to injured phenotype at approximately 35% Lagrangian strain. Continued development of this model may create novel opportunities for drug discovery and exploration of the role of human genotype in TAI pathology. PMID:27671211

  3. 96-well electroporation method for transfection of mammalian central neurons.

    PubMed

    Buchser, William J; Pardinas, Jose R; Shi, Yan; Bixby, John L; Lemmon, Vance P

    2006-11-01

    Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to obtain better efficiencies. We have developed an efficient method for the transfection of cerebellar granule neurons and hippocampal neurons with standard plasmid vectors. Using 96-well electroporation plates, square-wave pulses can introduce 96 different plasmids into neurons in a single step. The procedure results in greater than 20% transfection efficiencies and requires only simple solutions of nominal cost. In addition to enabling the rapid optimization of experimental protocols with multiple parameters, this procedure enables the use of high content screening methods to characterize neuronal phenotypes.

  4. 96-Well electroporation method for transfection of mammalian central neurons

    PubMed Central

    Buchser, William J.; Pardinas, Jose R.; Shi, Yan; Bixby, John L.; Lemmon, Vance P.

    2008-01-01

    Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to obtain better efficiencies. We have developed an efficient method for the transfection of cerebellar granule neurons and hippocampal neurons with standard plasmid vectors. Using 96-well electroporation plates, square-wave pulses can introduce 96 different plasmids into neurons in a single step. The procedure results in greater than 20% transfection efficiencies and requires only simple solutions of nominal cost. In addition to enabling the rapid optimization of experimental protocols with multiple parameters, this procedure enables the use of high content screening methods to characterize neuronal phenotypes. PMID:17140120

  5. An improved 96-well turbidity assay for T4 lysozyme activity

    PubMed Central

    Toro, Tasha B.; Nguyen, Thao P.; Watt, Terry J.

    2015-01-01

    T4 lysozyme (T4L) is an important model system for investigating the relationship between protein structure and function. Despite being extensively studied, a reliable, quantitative activity assay for T4L has not been developed. Here, we present an improved T4L turbidity assay as well as an affinity-based T4L expression and purification protocol. This assay is designed for 96-well format and utilizes conditions amenable for both T4L and other lysozymes. This protocol enables easy, efficient, and quantitative characterization of T4L variants and allows comparison between different lysozymes. Our method: • Is applicable for all lysozymes, with enhanced sensitivity for T4 lysozyme compared to other 96-well plate turbidity assays; • Utilizes standardized conditions for comparing T4 lysozyme variants and other lysozymes; and • Incorporates a simplified expression and purification protocol for T4 lysozyme. PMID:26150996

  6. 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.

  7. 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

  8. 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...

  9. 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...

  10. 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...

  11. 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.

  12. 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

  13. 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.

  14. Inoculation of culture plates: straightforward or is it?

    PubMed

    Carlin, E M; Hammond, J A; Boag, F C

    1997-04-01

    In view of the recent vogue in some genitourinary medicine (GUM) units towards selective microscopy we aimed to assess the adequacy of culture plate inoculation in our own GUM clinic by the visual examination of 350 consecutively inoculated plates. Seventy-five (21%) plates were inoculated so lightly that no indentation in the agar could be seen whilst in 20 (60%) the agar was shredded. Eighty-five per cent of inadequately plated samples were inoculated by the same staff members who were either relatively inexperienced, or well-distanced from their last in-service training. This has many important implications not only in the identification and control of infection but also with respect to staff training. We have now introduced practical plating instruction for all new members of clinical staff and additional in-service training. We plan to repeat the audit in 6 months' time to assess the effect of these changes.

  15. Microfluidic titer plate for stratified 3D cell culture.

    PubMed

    Trietsch, Sebastiaan J; Israëls, Guido D; Joore, Jos; Hankemeier, Thomas; Vulto, Paul

    2013-09-21

    Human tissues and organs are inherently heterogeneous. Their functionality is determined by the interplay between different cell types, their secondary architecture, vascular system and gradients of signaling molecules and metabolites. Here we propose a stratified 3D cell culture platform, in which adjacent lanes of gels and liquids are patterned by phaseguides to capture this tissue heterogeneity. We demonstrate 3D cell culture of HepG2 hepatocytes under continuous perfusion, a rifampicin toxicity assay and co-culture with fibroblasts. 4T1 breast cancer cells are used to demonstrate invasion and aggregation models. The platform is incorporated in a microtiter plate format that renders it fully compatible with automation and high-content screening equipment. The extended functionality, ease of handling and full compatibility to standard equipment is an important step towards adoption of Organ-on-a-Chip technology for screening in an industrial setting.

  16. 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.

  17. Enzyme activity assay of glycoprotein enzymes based on a boronate affinity molecularly imprinted 96-well microplate.

    PubMed

    Bi, Xiaodong; Liu, Zhen

    2014-12-16

    Enzyme activity assay is an important method in clinical diagnostics. However, conventional enzyme activity assay suffers from apparent interference from the sample matrix. Herein, we present a new format of enzyme activity assay that can effectively eliminate the effects of the sample matrix. The key is a 96-well microplate modified with molecularly imprinted polymer (MIP) prepared according to a newly proposed method called boronate affinity-based oriented surface imprinting. Alkaline phosphatase (ALP), a glycoprotein enzyme that has been routinely used as an indicator for several diseases in clinical tests, was taken as a representative target enzyme. The prepared MIP exhibited strong affinity toward the template enzyme (with a dissociation constant of 10(-10) M) as well as superb tolerance for interference. Thus, the enzyme molecules in a complicated sample matrix could be specifically captured and cleaned up for enzyme activity assay, which eliminated the interference from the sample matrix. On the other hand, because the boronate affinity MIP could well retain the enzymatic activity of glycoprotein enzymes, the enzyme captured by the MIP was directly used for activity assay. Thus, additional assay time and possible enzyme or activity loss due to an enzyme release step required by other methods were avoided. Assay of ALP in human serum was successfully demonstrated, suggesting a promising prospect of the proposed method in real-world applications.

  18. A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities

    PubMed Central

    Hao, Rui; Wei, Yuanchen; Li, Chaobo; Chen, Feng; Chen, Deyong; Zhao, Xiaoting; Luan, Shaoliang; Fan, Beiyuan; Guo, Wei; Wang, Junbo; Chen, Jian

    2017-01-01

    This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay. PMID:28240272

  19. A Microfabricated 96-Well 3D Assay Enabling High-Throughput Quantification of Cellular Invasion Capabilities.

    PubMed

    Hao, Rui; Wei, Yuanchen; Li, Chaobo; Chen, Feng; Chen, Deyong; Zhao, Xiaoting; Luan, Shaoliang; Fan, Beiyuan; Guo, Wei; Wang, Junbo; Chen, Jian

    2017-02-27

    This paper presents a 96-well microfabricated assay to study three-dimensional (3D) invasion of tumor cells. A 3D cluster of tumor cells was first generated within each well by seeding cells onto a micro-patterned surface consisting of a central fibronectin-coated area that promotes cellular attachment, surrounded by a poly ethylene glycol (PEG) coated area that is resistant to cellular attachment. Following the formation of the 3D cell clusters, a 3D collagen extracellular matrix was formed in each well by thermal-triggered gelation. Invasion of the tumor cells into the extracellular matrix was subsequently initiated and monitored. Two modes of cellular infiltration were observed: A549 cells invaded into the extracellular matrix following the surfaces previously coated with PEG molecules in a pseudo-2D manner, while H1299 cells invaded into the extracellular matrix in a truly 3D manner including multiple directions. Based on the processing of 2D microscopic images, a key parameter, namely, equivalent invasion distance (the area of invaded cells divided by the circumference of the initial cell cluster) was obtained to quantify migration capabilities of these two cell types. These results validate the feasibility of the proposed platform, which may function as a high-throughput 3D cellular invasion assay.

  20. Development and Testing of a Colorimetric 96 Well Plate Assay for the Determination of HD Hydrolysis Rate in Various Formulations

    DTIC Science & Technology

    2005-01-01

    Cold Foam 1% ethylene glycol 1% benzalkonium chloride 1% polyvinylpolypyrrolidone 1% Brij 58 1% Benzyldimethyl tetradecyl ammonium chloride dihydrate...LITERA TURE CITED ..................................................................................... 15 5 FIGURES 1. Rates of Chloride Release in...Ground, MD). The HD was stabilized with tributylamine and was approximately 90% pure. Quantitative assays were conducted with a chloride electrode

  1. Development of a rapid, 96-well alkaline based differential DNA extraction method for sexual assault evidence.

    PubMed

    Hudlow, William R; Buoncristiani, Martin R

    2012-01-01

    We present a rapid alkaline lysis procedure for the extraction of DNA from sexual assault evidence that generates purified sperm fraction extracts that yield STR typing results similar to those obtained from the traditional organic/dithiothreitol differential extraction. Specifically, a sodium hydroxide based differential extraction method has been developed in a single-tube format and further optimized in a 96-well format. The method yields purified extracts from a small sample set (≈ 2-6 swabs) in approximately 2h and from a larger sample set (up to 96 swabs) in approximately 4h. While conventional differential extraction methods require vigorous sample manipulation to remove the spermatozoa from the substrate, the method described here exploits the propensity of sperm to adhere to a substrate and does not require any manipulation of the substrate after it is sampled. For swabs, sample handling is minimized by employing a process where the tip of the swab, including the shaft, is transferred to the appropriate vessel eliminating the need for potentially hazardous scalpels to separate the swab material from the shaft. The absence of multiple handling steps allows the process to be semi-automated, however the procedure as described here does not require use of a robotic system. This method may provide forensic laboratories a cost-effective tool for the eradication of backlogs of sexual assault evidence, and more timely service to their client agencies. In addition, we have demonstrated that a modification of the procedure can be used to retrieve residual sperm-cell DNA from previously extracted swabs.

  2. Higher throughput bioanalysis by automation of a protein precipitation assay using a 96-well format with detection by LC-MS/MS.

    PubMed

    Watt, A P; Morrison, D; Locker, K L; Evans, D C

    2000-03-01

    Generic methodology for the automated preparation and analysis of drug levels in plasma samples within a drug discovery environment was achieved through the redesign of a protein precipitation assay to a microtiter (96-well) plate format and the application of robotic liquid handling for performance of all transfer and pipetting steps. Validation studies revealed that the application of robotics to sample preparation, in general, maintained the analytical accuracy and precision compared with preparing samples manually. The use of rapid gradient LC-MS/MS for analysis coupled with flow diversion of the solvent front allowed the introduction of protein-precipitated samples into the mass spectrometer without the necessity for source cleaning. The problem inherent in automatically pipetting plasma, caused by fibrinogen clots, was overcome by storing samples at -80 degrees C and thus precluding clot formation. The resulting methodology allowed sample preparation for a 96-well plate designed to accommodate 54 unknowns, duplicate 12-point calibration curves, and 6 sets of quality controls at three levels in approximately 2 h. This approach allowed an increase in throughput of sample preparation and analysis to >400 samples per day per LC-MS/MS instrument with minimal manual intervention. Overall, substantial time savings were realized, demonstrating that automation is an increasingly essential tool in a drug discovery bioanalytical environment.

  3. Analysis of a high-throughput cone-and-plate apparatus for the application of defined spatiotemporal flow to cultured cells.

    PubMed

    Spruell, Christopher; Baker, Aaron B

    2013-06-01

    The shear stresses derived from blood flow regulate many aspects of vascular and immunobiology. In vitro studies on the shear stress-mediated mechanobiology of endothelial cells have been carried out using systems analogous to the cone-and-plate viscometer in which a rotating, low-angle cone applies fluid shear stress to cells grown on an underlying, flat culture surface. We recently developed a device that could perform high-throughput studies on shear-mediated mechanobiology through the rotation of cone-tipped shafts in a standard 96-well culture plate. Here, we present a model of the three-dimensional flow within the culture wells with a rotating, cone-tipped shaft. Using this model we examined the effects of modifying the design parameters of the system to allow the device to create a variety of flow profiles. We first examined the case of steady-state flow with the shaft rotating at constant angular velocity. By varying the angular velocity and distance of the cone from the underlying plate we were able to create flow profiles with controlled shear stress gradients in the radial direction within the plate. These findings indicate that both linear and non-linear spatial distributions in shear stress can be created across the bottom of the culture plate. In the transition and "parallel shaft" regions of the system, the angular velocities needed to provide high levels of physiological shear stress (5 Pa) created intermediate Reynolds number Taylor-Couette flow. In some cases, this led to the development of a flow regime in which stable helical vortices were created within the well. We also examined the system under oscillatory and pulsatile motion of the shaft and demonstrated minimal time lag between the rotation of the cone and the shear stress on the cell culture surface.

  4. 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.

  5. Membrane-assisted culture of fungal mycelium on agar plates for RNA extraction and pharmacological analyses.

    PubMed

    Lange, Mario; Müller, Carolin; Peiter, Edgar

    2014-05-15

    Fungal mycelium grown in liquid culture is easy to harvest for RNA extraction and gene expression analyses, but liquid cultures often develop rather heterogeneously. In contrast, growth of fungal mycelium on agar plates is highly reproducible. However, this biological material cannot be harvested easily for downstream analyses. This article describes a PVDF (polyvinylidene difluoride) membrane-assisted agar plate culture method that enables the harvest of mycelium grown on agar plates. This culture method leads to a strongly reduced variation in gene expression between biological replicates and requires less growth space as compared with liquid cultures.

  6. 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

  7. A semi-automated 96-well protein precipitation method for the determination of montelukast in human plasma using high performance liquid chromatography/fluorescence detection.

    PubMed

    Kitchen, Chester J; Wang, Amy Q; Musson, Donald G; Yang, Amy Y; Fisher, Alison L

    2003-03-26

    A simple, semi-automated, protein precipitation assay for the determination of montelukast (SINGULAIR, MK-0476) in human plasma has been developed. Montelukast is a potent and selective antagonist of the cysteinyl leukotriene receptor used for the treatment of asthma. A Packard MultiPROBE II EX is used to transfer 300 microl of plasma from sample, standard, and QC sample tubes to a microtiter plate (96-well). After addition of the internal standard by a repeating pipettor, a Tomtec QUADRA 96 adds 400 microl of acetonitrile to all plasma sample wells, simultaneously, in the microtiter plate. The Tomtec is also used to transfer the acetonitrile supernatant from the plasma protein precipitation step, batchwise, to another microtiter plate for analysis by HPLC with fluorescence detection. This assay has been validated and implemented for a clinical study of over 1300 plasma samples and is comparable to manual assays in the LLOQ (lower limit of quantitation, 3 ng/ml) and in stability. This is the first semi-automated protein precipitation assay published for the analysis of montelukast in human plasma and it results in significant time savings over the manual methods, both in sample preparation and in HPLC run time.

  8. 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...

  9. 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.

  10. Simplified method for determination of clarithromycin in human plasma using protein precipitation in a 96-well format and liquid chromatography-tandem mass spectrometry.

    PubMed

    Shin, Jaekyu; Pauly, Daniel F; Johnson, Julie A; Frye, Reginald F

    2008-08-01

    A simplified method to determine clarithromycin concentrations in human plasma using protein precipitation in a 96-well plate and liquid chromatography-tandem mass spectrometry was developed and validated. Plasma proteins were precipitated with acetonitrile and roxithromycin was used as the internal standard. After vortex mixing and centrifugation, the supernatants were directly injected onto a Phenomenex Luna Phenyl-Hexyl column (50 mm x 2.0 mm ID, 3 microm). The mobile phase consisted of water and methanol (30:70, v/v) containing 0.1% formic acid and 5mM ammonium acetate. The flow rate was 0.22 mL/min and the total run time (injection to injection) was less than 3 min. Detection of the analytes was achieved using positive ion electrospray tandem mass spectrometry in selected reaction monitoring (SRM) mode. The linear standard curve ranged from 100 to 5000 ng/mL and the precision and accuracy (inter- and intra-run) were within 7.9% and 4.9%, respectively. The method was successfully used to determine clarithromycin concentrations in human plasma samples obtained from healthy subjects who were given clarithromycin 500 mg for 3 days. The method is rapid, simple, precise and directly applicable to clarithromycin pharmacokinetic studies.

  11. Autogenous cultured growth plate chondrocyte transplantation in the treatment of physeal injury in rabbits

    PubMed Central

    Tomaszewski, R.; Bohosiewicz, J.; Gap, A.; Bursig, H.; Wysocka, A.

    2014-01-01

    Objectives The aim of this experimental study on New Zealand’s white rabbits was to investigate the transplantation of autogenous growth plate cells in order to treat the injured growth plate. They were assessed in terms of measurements of radiological tibial varus and histological characteristics. Methods An experimental model of plate growth medial partial resection of the tibia in 14 New Zealand white rabbits was created. During this surgical procedure the plate growth cells were collected and cultured. While the second surgery was being performed, the autologous cultured growth plate cells were grafted at the right tibia, whereas the left tibia was used as a control group. Results Histological examinations showed that the grafted right tibia presented the regular shape of the plate growth with hypertrophic maturation, chondrocyte columniation and endochondral calcification. Radiological study shows that the mean tibial deformity at the left angle was 20.29° (6.25 to 33) and 7.21° (5 to 10) in the right angle. Conclusion This study has demonstrated that grafting of autogenous cultured growth plate cells into a defect of the medial aspect of the proximal tibial physis can prevent bone bridge formation, growth arrest and the development of varus deformity. Cite this article: Bone Joint Res 2014;3:310–16 PMID:25376625

  12. Strongyloidiasis detected by the agar plate culture method among patients infected by HIV.

    PubMed

    Urdez-Hernández, E; Jiménez-Galán, S; Antonio-Manríquez, M; DE León-Juárez, E A; Terrazas-Estrada, J J; Hernández-García, M C; García-Zaldívar, P; Estrada-Aguilera, A

    1999-10-01

    To evaluate the rate of strongyloidiasis among HIV/AIDS patients, stools and duodenal juice were examined using the agar plate culture method. From January to June 1993, a total of 60 HIV/AIDS patients were required for duodenal aspirate and three serial samples of freshly passed stools. Stools and duodenal aspirate were dispensed on an agar plate culture; after incubation at 28 degrees C during 48 h, screening of plates was made at 10 x. The presence of furrows and worms of short buccal chamber and prominent genital primordium were positive for Strongyloides stercoralis. Most patients were men (91.7%); their mean age, of 33.9 years (SD +/- 10.6); their median CD4(+) T-cells count, of 105/microL (range of 12 to 646). S. stercoralis was detected in three patients (5%). In duodenal juice, the three patients showed the parasite, but in feces, only two (3.3%). In these two individuals, the worms were found in feces by agar culture and Faust's concentration method. The rate of S. stercoralis in feces of HIV/AIDS individuals (3.3%) by agar culture method was similar to that formerly reported from the general Mexican population (2.9%) using standard concentration procedures. Hence, in this immunocompromised population of a low prevalence city, there was no advantage to using an agar plate culture for strongyloidiasis.

  13. EFFECTS OF PLATING DENSITY AND CULTURE TIME ON BONE MARROW STROMAL CELL CHARACTERISTICS

    PubMed Central

    Neuhuber, Birgit; Swanger, Sharon A.; Howard, Linda; Mackay, Alastair; Fischer, Itzhak

    2008-01-01

    Objective Bone marrow stromal cells (MSC) are multipotent adult stem cells that have emerged as promising candidates for cell therapy in disorders including cardiac infarction, stroke and spinal cord injury. While harvesting methods used by different laboratories are relatively standard, MSC culturing protocols vary widely. This study is aimed at evaluating the effects of initial plating density and total time in culture on proliferation, cell morphology, and differentiation potential of heterogeneous MSC cultures and more homogeneous cloned subpopulations. Methods Rat MSC were plated at 20, 200 and 2000 cells/cm2 and grown to 50% confluency. The numbers of population doublings and doubling times were determined within and across multiple passages. Changes in cell morphology and differentiation potential to adipogenic, chondrogenic, and osteogenic lineages were evaluated and compared among early, intermediate and late passages, as well as between heterogeneous and cloned MSC populations. Results We found optimal cell growth at a plating density of 200 cells/cm2. Cultures derived from all plating densities developed increased proportions of flat cells over time. Assays for chondrogenesis, osteogenesis and adipogenesis showed that heterogeneous MSC plated at all densities sustained the potential for all three mesenchymal phenotypes through at least passage 5; the flat subpopulation lost adipogenic and chondrogenic potential. Conclusion Our findings suggest that the initial plating density is not critical for maintaining a well-defined, multipotent MSC population. Time in culture, however, affects cell characteristics, suggesting that cell expansion should be limited, especially until the specific characteristics of different MSC subpopulations are better understood. PMID:18495329

  14. Phospholipid removal combined with a semi-automated 96-well SPE application for determination of budesonide in human plasma with LC-MS/MS.

    PubMed

    Nilsson, Karin; Andersson, Maria; Beck, Olof

    2014-11-01

    A semi-automated method for quantification of budesonide in human plasma was developed, validated, and applied for high-volume analysis of samples in connection with a pharmacokinetic study. Protein and phospholipid removal was performed using an Ostro 96-well filter plate and subsequently combined with C18 solid-phase extraction on a Hamilton Microlab STARlet automation robot. The final extracts were evaporated to dryness and redissolved in 20% acetonitrile/water. The procedure used budesonide-d8 as internal standard and gave a 3.5-fold concentration of plasma to extract. The final extracts (5 μL injected) were analyzed with selected reaction monitoring liquid chromatography-tandem mass spectrometry (LC-MS/MS) using electrospray ionization in positive mode. The chromatography system used a 100 mm ACQUITY BEH UPLC column and a gradient system consisting of aqueous 0.1% formic acid and acetonitrile as organic modifier. Phospholipid removal was found to be needed during method development in order to reduce ion suppression effects from matrix and to increase method sensitivity. The measuring range was 50-5000 pg/mL with and LOD 24 pg/mL. Calibration response showed good linearity (correlation coefficients<0.99) over the measuring range. The absolute recovery over the sample preparation procedure was estimated to 67%. Total imprecision was <9% at three levels and accuracy was between 98.9 and 103%. The method was successfully applied for analysis of 864 study samples in a short time. The quality control samples at concentration levels 200 and 2000 pg/mL gave a total imprecision of 7.4% and 4.2%, respectively, (n=95).

  15. Accumulation of neurons differentiated from mouse embryonic stem cells in particular areas of culture plate surface.

    PubMed

    Kitazawa, Ayako; Naka, Yukie; Yamaguchi, Hiroko; Shimizu, Norio

    2010-08-01

    Nanoscale magnetic beads coated with nerve growth factor (NGF) allow us to accumulate neurons differentiated from mouse ES cells in a selected area of the culture plate surface using a magnet. Neurons with neurite outgrowths within a particular area expressed TrkA and incorporated beads in the soma.

  16. A 96-well single-pot liquid-liquid extraction, hydrophilic interaction liquid chromatography-mass spectrometry method for the determination of muraglitazar in human plasma.

    PubMed

    Xue, Y-J; Liu, Jane; Unger, Steve

    2006-06-07

    A single-pot liquid-liquid extraction (LLE) with hydrophilic interaction liquid chromatography/tandem mass spectrometry (HILIC/MS/MS) method has been developed and validated for the determination of muraglitazar, a hydrophobic diabetes drug, in human plasma. To 0.050 ml of each plasma sample in a 96-well plate, the internal standard solution in acetonitrile and toluene were added to extract the compound of interest. The plate was vortexed, followed by centrifugation. The organic layer was then directly injected into an LC/MS/MS system. Chromatographic separation was achieved isocratically on a Thermohypersil_Keystone, Hypersil silica column (3 mmx50 mm, 3 microm). The mobile phase contained 85% of methyl t-butyl ether and 15% of 90/10 (v/v) acetonitrile/water with 0.3% trifluoroacetic acid. Post-column mobile phase of 50/50 (v/v) acetonitrile/water containing 0.1% formic acid was added. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API 4000. The standard curve, ranged from 1 to 1000 ng/ml, was fitted to a 1/x weighted quadratic regression model. This single-pot LLE approach effectively eliminated time-consuming organic layer transfer, dry-down, and sample reconstitution steps, which are essential for a conventional liquid-liquid extraction procedure. The modified mobile phase was more compatible with the direct injection of the commonly used extraction solvents in LLE. Furthermore, the modified mobile phase improved the retention of muraglitazar, a hydrophobic compound, on the normal phase silica column. The validation results demonstrated that this method was rugged and suitable for analyzing muraglitazar in human plasma. In comparison with a revised-phase LC/MS/MS method, this single-pot LLE, HILIC/MS/MS method improved the detection sensitivity by more than four-fold based upon the LLOQ signal to noise ratio. This approach may be applied to other hydrophobic compounds with proper modification of the mobile phase compositions.

  17. Increased sensitivity of routine laboratory detection of Strongyloides stercoralis and hookworm by agar-plate culture.

    PubMed

    Jongwutiwes, S; Charoenkorn, M; Sitthichareonchai, P; Akaraborvorn, P; Putaporntip, C

    1999-01-01

    The efficacy of agar-plate culture has been evaluated for the detection of Strongyloides stercoralis and hookworm, compared with direct smear, the formalin-ether sedimentation technique and the filter-paper method. Of 1085 stool samples from the routine laboratory service at King Chulalongkorn Memorial Hospital in Bangkok, 241 samples harboured S. stercoralis, 153 hookworm and 2 Rhabditis hominis. The recovery rate of S. stercoralis by agar-plate culture is significantly superior to the other methods (P < 0.005). The ratios of positive results from the methods used to the total number of S. stercoralis-positive cases were as follows: 1:1.03 by agar-plate culture, 1:1.85 by the filter-paper method, 1:1.98 by the sedimentation technique and 1:10.48 by direct stool smear. A similar trend of the efficacy ratio of each method was obtained for hookworm detection. The characteristic furrows left by hookworm larvae, and larvae and adults of S. stercoralis could be used for preliminary species identification. Daily search for furrows on agar plates for up to 6 consecutive days resulted in an increased sensitivity for diagnosis of both S. stercoralis and hookworm infections.

  18. Fluorescence assay for mitochondrial permeability transition in cardiomyocytes cultured in a microtiter plate.

    PubMed

    Christensen, Marie Louise Muff; Braunstein, Thomas Hartig; Treiman, Marek

    2008-07-01

    Mitochondrial permeability transition pore (MPTP) is a voltage-dependent, large-conductance channel of the inner mitochondrial membrane with an important role in a range of pathophysiological conditions. To facilitate studies of pharmacological pore modulation, we describe an assay in a model using neonatal cardiomyocytes in a 96-well microtiter plate format. In the presence of mitochondrial membrane potential Delta Psi m, accumulation of rhodamine-123 in mitochondria (40,000 cells/well, 2.6 microM rhodamine-123) caused fluorescence signal quenching. Following substitution of dye-free buffer, dequenching occurred on the distribution of rhodamine-123 into the extracellular volume. The addition of a small buffer volume containing digitonin (final concentration 10 microg/ml) and Ca(2+) (final concentrations up to 100 microM free Ca(2+)) caused dequenching (Delta F) due to Delta Psi m dissipation by MPTP, as evidenced by inhibition in the presence of cyclosporin A (0.2-2 microM) and facilitation by pH 6.2. Delta F due to Delta Psi m-dissipating agent carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) or alamethicin (10 microM) was insensitive to either pH or cyclosporin A. Inhibition of Ca(2+)-induced (but not of FCCP- or alamethicin-induced) Delta F by glycogen synthase kinase 3beta (GSK3 beta) antagonist SB216763 and adenosine, acting at the level of intracellular signaling and plasma membrane receptors, respectively, is shown to illustrate potential applications of this assay. Limitation of the assay to cells with energized mitochondria is stressed.

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

    PubMed

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

    2014-07-01

    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.

  20. 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

    2016-05-19

    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.

  1. 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

  2. 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.

  3. A simple 96-well liquid-liquid extraction with a mixture of acetonitrile and methyl t-butyl ether for the determination of a drug in human plasma by high-performance liquid chromatography with tandem mass spectrometry.

    PubMed

    Xue, Y-J; Pursley, Janice; Arnold, Mark E

    2004-02-04

    A simple 96-well plate liquid-liquid extraction (LLE), liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of a basic drug candidate in human plasma. Against the wisdom of conventional approaches, an aqueous/organic miscible solvent, acetonitrile, was used for liquid-liquid extraction along with methyl t-butyl ether. The use of acetonitrile effectively eliminated the formation of the irregular emulsion between aqueous/organic interfaces and modulated the polarity of the extraction solvents to achieve the desired recovery. This approach, which solved the emulsion problem, permitted the method to be automated using standard 96-well plate technology. A practical application was demonstrated through the use of this technique in the measurement of a novel drug in human plasma samples by LC/MS/MS. Chromatographic separation was achieved isocratically on a Phenomenox C18(2) Luna column (2 mm x 50 mm, 5 microm). The mobile phase contained 60% of 0.1% formic acid and 40% acetonitrile. Detection was by positive ion electrospray tandem mass spectrometry. The standard curve, which ranged from 1.22 to 979ng/ml, was fitted to a 1/x2 weighted quadratic regression model. The validation results show that this method was very rugged and had excellent precision and accuracy. The actual sample analysis results further demonstrated that this extraction procedure is well suited for real life applications. It is expected that with some modifications, this approach can be applied for the extraction of similar compounds from various biological fluids.

  4. A 96-well single-pot protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of muraglitazar, a novel diabetes drug, in human plasma.

    PubMed

    Xue, Y-J; Liu, Jane; Pursley, Janice; Unger, Steve

    2006-02-02

    A 96-well single-pot protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of muraglitazar, a PPAR alpha/gamma dual agonist, in human plasma. The internal standard, a chemical analogue, was dissolved in acetonitrile containing 0.1% formic acid. The solvent system was also served as a protein precipitation reagent. Human plasma samples (0.1 mL) and the internal standard solution (0.3 mL) were added to a 96-well plate. The plate was vortexed for 1 min and centrifuged for 5 min. Then the supernatant layers were directly injected into the LC/MS/MS system. The chromatographic separation was achieved isocratically on a Phenomenox C18(2) Luna column (2 mm x 50 mm, 5 microm). The mobile phase contained 20/80 (v/v) of water and acetonitrile containing 0.1% formic acid. Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API 3000. The standard curve, which ranged from 1 to 1000 ng/mL, was fitted to a 1/x weighted quadratic regression model. This single-pot approach effectively eliminated three time consuming sample preparation steps: sample transfer, dry-down, and reconstitution before the injection, while it preserved all the benefits of the traditional protein precipitation. By properly adjusting the autosampler needle offset level, only the supernatant was injected, without disturbing the precipitated proteins in the bottom. As a result, the quality of chromatography and column life were not compromised. After more than 600 injections, there was only slightly increase of column back-pressure. The validation results demonstrated that this method was rugged and provide satisfactory precision and accuracy. The method has been successfully applied to analyze human plasma samples in support of a first-in-man study. This method has also been validated in monkey and mouse plasma for the determination of muraglitazar.

  5. 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

  6. Gene Transfection toward Spheroid Cells on Micropatterned Culture Plates for Genetically-modified Cell Transplantation.

    PubMed

    Itaka, Keiji; Uchida, Satoshi; Matsui, Akitsugu; Yanagihara, Kayoko; Ikegami, Masaru; Endo, Taisuke; Ishii, Takehiko; Kataoka, Kazunori

    2015-07-31

    To improve the therapeutic effectiveness of cell transplantation, a transplantation system of genetically modified, injectable spheroids was developed. The cell spheroids are prepared in a culture system on micropatterned plates coated with a thermosensitive polymer. A number of spheroids are formed on the plates, corresponding to the cell adhesion areas of 100 µm diameter that are regularly arrayed in a two-dimensional manner, surrounded by non-adhesive areas that are coated by a polyethylene glycol (PEG) matrix. The spheroids can be easily recovered as a liquid suspension by lowering the temperature of the plates, and their structure is well maintained by passing them through injection needles with a sufficiently large caliber (over 27 G). Genetic modification is achieved by gene transfection using the original non-viral gene carrier, polyplex nanomicelle, which is capable of introducing genes into cells without disrupting the spheroid structure. For primary hepatocyte spheroids transfected with a luciferase-expressing gene, the luciferase is sustainably obtained in transplanted animals, along with preserved hepatocyte function, as indicated by albumin expression. This system can be applied to a variety of cell types including mesenchymal stem cells.

  7. Well Plate-Based Perfusion Culture Device for Tissue and Tumor Microenvironment Replication

    PubMed Central

    Zhang, W.; Gu, Y.; Hao, Y.; Sun, Q.; Konior, K.; Wang, H.

    2015-01-01

    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) 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

  8. 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

    2016-01-01

    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.

  9. Comparison of dry medium culture plates for mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

    PubMed

    Park, Junghyun; Kim, Myunghee

    2013-12-01

    This study was performed to compare the performance of Sanita-Kun dry medium culture plate with those of traditional culture medium and Petrifilm dry medium culture plate for the enumeration of the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet. Mesophilic aerobic bacteria were comparatively evaluated in milk, ice cream, ham, and codfish fillet using Sanita-Kun aerobic count (SAC), Petrifilm aerobic count (PAC), and traditional plate count agar (PCA) media. According to the results, all methods showed high correlations of 0.989~1.000 and no significant differences were observed for enumerating the mesophilic aerobic bacteria in the tested food products. SAC method was easier to perform and count colonies efficiently as compared to the PCA and PAC methods. Therefore, we concluded that the SAC method offers an acceptable alternative to the PCA and PAC methods for counting the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products.

  10. Device for the control of oxygen concentration in multiwell cell culture plates.

    PubMed

    Oppegard, Shawn C; Eddington, David T

    2009-01-01

    Oxygen is a key modulator of many cellular pathways but current devices permitting in vitro oxygen modulation fail to meet the needs of many researchers. In this study, a microfabricated insert for multiwell formats has been developed to control the gas concentration of each well independent of the global incubator's condition. The platform consists of a polydimethylsiloxane (PDMS) insert that nests into a standard multiwell plate and serves as a passive network with a gas permeable membrane aimed to deliver gas to adherent cell cultures. Preliminary data demonstrate that the insert is effective in controlling the oxygen concentration at the cell surface inside a well with equilibration times in minutes rather than hours for conventional technologies. A wide variety of oxygen profiles can be attained based on the device design, such as the cyclic profile achieved in this study, and even gradients in local oxygen concentration to mimic those found in vivo for more biomimetic cellular models.

  11. Automated 96-well solid phase extraction and hydrophilic interaction liquid chromatography-tandem mass spectrometric method for the analysis of cetirizine (ZYRTEC) in human plasma--with emphasis on method ruggedness.

    PubMed

    Song, Qi; Junga, Heiko; Tang, Yong; Li, Austin C; Addison, Tom; McCort-Tipton, Melanie; Beato, Brian; Naidong, Weng

    2005-01-05

    A high-throughput bioanalytical method based on automated sample transfer, automated solid phase extraction, and hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) analysis, has been developed for the determination of cetirizine, a selective H(1)-receptor antagonist. Deuterated cetirizine (cetirizine-d(8)) was synthesized as described and was used as the internal standard. Samples were transferred into 96-well plates using an automated sample handling system. Automated solid phase extraction was carried out using a 96-channel programmable liquid-handling workstation. Solid phase extraction 96-well plate on polymer sorbent (Strata X) was used to extract the analyte. The extracted samples were injected onto a Betasil silica column (50 x 3, 5 microm) using a mobile phase of acetonitrile-water-acetic acid-trifluroacetic acid (93:7:1:0.025, v/v/v/v) at a flow rate of 0.5 ml/min. The chromatographic run time is 2.0 min per injection, with retention time of cetirizine and cetirizine-d(8) both at 1.1 min. The system consisted of a Shimadzu HPLC system and a PE Sciex API 3000 or API 4000 tandem mass spectrometer with (+) ESI. The method has been validated over the concentration range of 1.00-1000 ng/ml cetirizine in human plasma, based on a 0.10-ml sample size. The inter-day precision and accuracy of the quality control (QC) samples demonstrated <3.0% relative standard deviation (R.S.D.) and <6.0% relative error (RE). Stability of cetirizine in stock solution, in plasma, and in reconstitution solution was established. The absolute extraction recovery was 85.8%, 84.5%, and 88.0% at 3, 40, and 800 ng/ml, respectively. The recovery for the internal standard was 84.1%. No adverse matrix effects were noticed for this assay. The automation of the sample preparation steps not only increased the analysis throughput, but also increased method ruggedness. The use of a stable isotope-labeled internal standard further improved the method ruggedness

  12. 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

  13. Comparison of microtitre plates with flat-bottomed and round-bottomed wells for mixed lymphocyte culture (MLC).

    PubMed

    Herva, E

    1977-04-01

    To compare microtitre plates with flat-bottomed and round-bottomed wells and to standardize a method for mixed lymphocyte culture (MLC), the effects of cell number, culture time, 3H-thymidine concentration and labelling time were studied. On both plates, allogeneic cells induced increased RNA synthesis beginning at about 30 hours and increased DNA synthesis beginning at about 50 hours, if suitable cell numbers were used. On plates with flat-bottomed wells, 1.5 X 10(5) responding and stimulating cells per well had near-exponential growth on day four and five, often through day six; on plates with round-bottomed wells the corresponding cell number was 0.25-1.0 (optimally 0.5) X 10(5). Near these cell numbers, the response depended closely on the number of responding cells. On plates with flat-bottomed wells, stimulating cells had a dose-dependent effect on the response, whereas on plates with round-bottomed wells, increasing the stimulating cell dose did not consistently strengthen the response. Both plate types proved suitable for MLC experiments; plates with round-bottomed wells have the obvious advantage of requiring smaller cell numbers. 3H-thymidine (spec, act 2000 mCi/mmol) self-suppressed its incorporation, which increased only slightly or even decreased if labelling time exceeded 12-18 hours. Relative responses remained virtually unaltered, however, with 3H-concentrations of 0.5 and 2.0 micronCi/ml and with labelling times of 8 and 24 hours.

  14. Surface functionalization of tissue culture polystyrene plates with hydroxyapatite under body fluid conditions and its effect on differentiation behaviors of mesenchymal stem cells.

    PubMed

    Iijima, Kazutoshi; Suzuki, Ryo; Iizuka, Ayako; Ueno-Yokohata, Hitomi; Kiyokawa, Nobutaka; Hashizume, Mineo

    2016-11-01

    The surfaces of polystyrene (PS) cell culture plates were functionalized with hydroxyapatite (HAp) under body fluid conditions utilizing protein adsorption layers and a pretreatment with an alternate soaking process (ASP) using solutions containing calcium and phosphate ions. Adsorption layers of human serum albumin (HSA) formed on the surface of each well of commercial 24-well PS plates by solution processes. CaCl2 and K2HPO4 solutions were alternately added to the wells, the plates were incubated to form the precursors, and this was followed by the addition of simulated body fluid (SBF) and a further incubation for 24h. These treatments resulted in the surfaces of the PS cell culture plates being completely covered with bone-like HAp. The coating of PS plates with HAp promoted the adhesion of mesenchymal stem cells (MSCs) and maintained cell growth that was as fast as that on tissue culture-treated PS (TCPS) plates. Osteogenic differentiation was greater, whereas adipogenic and chondrogenic differentiation was less in the culture on HAp-coated PS plates than in that on TCPS plates. The present method is useful for preparing HAp-coated PS plates at clean benches without the need for any expensive apparatus. HAp coated on PS plates by this method was a bone-like apatite with high bioactivity; therefore, the present HAp-coated PS plates are promising materials for assays of bone-related cells in the bone remodeling process.

  15. Enumeration of bacteria in clinically significant blood cultures in neutropenic and non-neutropenic patients using a pour plate method.

    PubMed

    Rice, P; Spencer, R C

    1991-03-01

    A 3-year review of clinically significant positive blood cultures was undertaken to assess any differences in the blood bacterial count between haematological neutropenic and other non-neutropenic patients. The pour-plate method was used. In Gram-positive infections the pour plate contained colonies in 61% of haematological patients and in 41% of others. In Gram-negative infection the figures were 54% and 25% respectively. The mean numbers of bacteria per ml of blood were increased in haematological patients compared with the others for both groups of organisms.

  16. 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.

  17. 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.

  18. Micro-Ring Structures Stabilize Microdroplets to Enable Long Term Spheroid Culture in 384 Hanging Drop Array Plates

    PubMed Central

    Hsiao, Amy Y.; Tung, Yi-Chung; Kuo, Chuan-Hsien; Mosadegh, Bobak; Bedenis, Rachel; Pienta, Kenneth J.; Takayama, Shuichi

    2012-01-01

    Using stereolithography, 20 different structural variations comprised of millimeter diameter holes surrounded by trenches, plateaus, or micro-ring structures were prepared and tested for their ability to stably hold arrays of microliter sized droplets within the structures over an extended period of time. The micro-ring structures were the most effective in stabilizing droplets against mechanical and chemical perturbations. After confirming the importance of micro-ring structures using rapid prototyping, we developed an injection molding tool for mass production of polystyrene 3D cell culture plates with an array of 384 such micro-ring surrounded through-hole structures. These newly designed and injection molded polystyrene 384 hanging drop array plates with micro-rings were stable and robust against mechanical perturbations as well as surface fouling-facilitated droplet spreading making them capable of long term cell spheroid culture of up to 22 days within the droplet array. This is a significant improvement over previously reported 384 hanging drop array plates which are susceptible to small mechanical shocks and could not reliably maintain hanging drops for longer than a few days. With enhanced droplet stability, the hanging drop array plates with micro-ring structures provide better platforms and open up new opportunities for high-throughput preparation of microscale 3D cell constructs for drug screening and cell analysis. PMID:22057945

  19. Method development for the quantitation of ABT-578 in rabbit artery tissue by 96-well liquid-liquid extraction and liquid chromatography/tandem mass spectrometric detection.

    PubMed

    Ji, Qin C; Zhang, Jun; Rodila, Ramona; Watson, Pamela; El-Shourbagy, Tawakol

    2004-01-01

    Quantitative determination of drug concentrations in tissue samples can provide critical information for drug metabolism, kinetics, and toxicity evaluations. For analysis of tissue samples using liquid chromatography/tandem mass spectrometric (LC/MS/MS) detection, homogenization is a critical step in achieving good assay performance. Assay performance can be closely evaluated by spiking the drug directly into tissue samples prior to homogenization. It is especially important to include this assay evaluation for the analysis of artery tissue samples because artery tissue is very elastic, making it quite a challenge to develop an effective procedure for homogenization. An LC/MS/MS assay in 96-well format using liquid-liquid extraction was developed for analyzing ABT-578 in rabbit artery samples. Tissue quality control samples were prepared by spiking ABT-578 stock solutions directly into the tissue before homogenization. The usage of the tissue control samples gives a thorough evaluation of the sample preparation process that includes both homogenization and sample extraction. A 20% blood in saline solution was used as a homogenization solution. Calibration standards were made by spiking ABT-578 into rabbit whole blood. Blood quality control samples were also prepared by spiking ABT-578 into rabbit whole blood. These blood QC samples were used to confirm the validity of the calibration curve. A lower limit of quantitation of 0.050 ng/mL was achieved. The linear dynamic range of blood standards was from 0.050-30.3 ng/mL with the correlation coefficient (r) ranging from 0.9969-0.9996. Overall %CV was between 1.3 and 7.0%, and analytical recovery was between 98.2 and 105.8% for blood QC samples. The %CVs for tissue QC samples were between 6.7 and 13.0%, and analytical recovery after correction was between 93.5 and 114.3%.

  20. A high-throughput protein refolding screen in 96-well format combined with design of experiments to optimize the refolding conditions.

    PubMed

    Dechavanne, Vincent; Barrillat, Nicolas; Borlat, Frederic; Hermant, Aurélie; Magnenat, Laurent; Paquet, Mikael; Antonsson, Bruno; Chevalet, Laurent

    2011-02-01

    Production of correctly folded and biologically active proteins in Escherichiacoli can be a challenging process. Frequently, proteins are recovered as insoluble inclusion bodies and need to be denatured and refolded into the correct structure. To address this, a refolding screening process based on a 96-well assay format supported by design of experiments (DOE) was developed for identification of optimal refolding conditions. After a first generic screen of 96 different refolding conditions the parameters that produced the best yield were further explored in a focused DOE-based screen. The refolding efficiency and the quality of the refolded protein were analyzed by RP-HPLC and SDS-PAGE. The results were analyzed by the DOE software to identify the optimal concentrations of the critical additives. The optimal refolding conditions suggested by DOE were verified in medium-scale refolding tests, which confirmed the reliability of the predictions. Finally, the refolded protein was purified and its biological activity was tested in vitro. The screen was applied for the refolding of Interleukin 17F (IL-17F), stromal-cell-derived factor-1 (SDF-1α/CXCL12), B cell-attracting chemokine 1 (BCA-1/CXCL13), granulocyte macrophage colony stimulating factor (GM-CSF) and the complement factor C5a. This procedure identified refolding conditions for all the tested proteins. For the proteins where refolding conditions were already available, the optimized conditions identified in the screening process increased the yields between 50% and 100%. Thus, the method described herein is a useful tool to determine the feasibility of refolding and to identify high-yield scalable refolding conditions optimized for each individual protein.

  1. Development, validation and application of a 96-well enzymatic assay based on LC-ESI-MS/MS quantification for the screening of selective inhibitors against Mycobacterium tuberculosis purine nucleoside phosphorylase.

    PubMed

    Cattaneo, Giulia; Ubiali, Daniela; Calleri, Enrica; Rabuffetti, Marco; Höfner, Georg C; Wanner, Klaus T; De Moraes, Marcela C; Martinelli, Leonardo K B; Santos, Diógenes Santiago; Speranza, Giovanna; Massolini, Gabriella

    2016-11-02

    Mycobacterium tuberculosis (Mtb) purine nucleoside phosphorylase (PNP, EC 2.4.2.1) has been identified as a target for the development of specific inhibitors with potential antimycobacterial activity. We hereby described the development and validation of a new 96-well LC-ESI-MS/MS method to assess the inhibition activity of nucleoside analogues towards MtbPNP and the human PNP (HsPNP). Enzyme activity was determined by monitoring the phosphorolysis of inosine (Ino) to hypoxanthine (Hpx). The enzymatic assay (v = 0.5 mL, enzyme<0.2 μg/well, T = 37 °C) was performed with an overall time of about 15 min/plate for sample processing and 2 min/sample for LC-MS analysis. Validation of the quantification method met the criteria of the CDER guidance of FDA. Kinetic parameters were in agreement with those reported in literature (HsPNP KM = 0.150 ± 0.020 mM vs 0.133 ± 0.015 mM; MtbPNP KM = 0.060 ± 0.009 mM vs 0.040 ± 0.003 mM for Ino), thus demonstrating the reliability of the newly developed enzymatic assay. Preliminary inhibition assays confirmed the effects reported for Acyclovir (Acv) and Formycin A (FA) against HsPNP and MtbPNP. The validated enzymatic assay was applied to the evaluation of a set of 8-halo-, 8-amino-, 8-O-alkyl-substituted purine ribonucleosides synthesized on purpose as potential inhibitors against MtbPNP. The assayed 8-substituted ribonucleosides did not exert a significant inhibitory effect against the tested enzymes up to 1 mM.

  2. In vitro maturation system for individual culture of bovine oocytes using micro-volume multi-well plate.

    PubMed

    Nagano, Masashi; Kang, Sung-Sik; Koyama, Keisuke; Huang, Weiping; Yanagawa, Yojiro; Takahashi, Yoshiyuki

    2013-11-01

    As a preliminary study for the development of individual in vitro maturation (IVM) culture of bovine oocytes, a multi-well (MW) plate was used. Maturation, fertilization and development to blastocysts were examined and compared with those of IVM oocytes cultured in 50-microl droplets in groups and in 10-microl droplets individually. The maturation rates were similar in all experimental groups. Normal fertilization rates in MW and 50-microl droplets were similar, but lower in 10-microl droplets (p < 0.01). The blastocyst rate in 10-microl droplets tended to be lower than those in MW (p = 0.15) and 50-microl droplets (p = 0.19). These results indicate that an IVM system using MW supports the acquisition of developmental competence by bovine oocytes the same as conventional group IVM culture.

  3. 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.

  4. Evaluation of three-dimensional cultured HepG2 cells in a nano culture plate system: an in vitro human model of acetaminophen hepatotoxicity.

    PubMed

    Aritomi, Kohei; Ishitsuka, Yoichi; Tomishima, Yoshiro; Shimizu, Daisuke; Abe, Nazuki; Shuto, Tsuyoshi; Irikura, Mitsuru; Kai, Hirofumi; Irie, Tetsumi

    2014-01-01

    Overdoses of acetaminophen (paracetamol, N-acetyl-p-aminophenol; APAP) cause severe liver injury, yet there is no common or high throughput in vitro human APAP model. This study examined the characteristics and usefulness of HepG2 cells grown in a nano culture plate (NCP) system, a three-dimensional culture method, as an in vitro human model for APAP-induced hepatotoxicity. The NCP-cultured HepG2 cells showed higher expression of mRNA and protein levels of cytochrome P450 2E1, which metabolizes APAP to a toxic metabolite, APAP-cysteine adduct formation, and higher sensitivity against APAP-induced cell injury compared with conventionally cultured cells. We demonstrated that treatment of APAP in NCP-cultured HepG2 cells shows key mechanistic features of APAP-induced hepatotoxicity, such as decreases in intracellular glutathione and mitochondrial membrane potential, activation of JNK, and cellular injury; and pharmacological agents, such as Cyclosporine A (a mitochondrial permeability transition inhibitor) and SP600125 (a JNK inhibitor), prevented cell injury induced by APAP exposure. In addition, the antidote of APAP-induced hepatotoxicity, N-acetylcysteine, could attenuate cellular injury induced by APAP in NCP-cultured HepG2 cells. We suggest that cellular injury induced by APAP treatment using an NCP-HepG2 system is a useful human model to study mechanisms and screen drug candidates of APAP-induced hepatotoxicity.

  5. Comparison of broth enhancement to direct plating for screening of rectal cultures for ciprofloxacin-resistant Escherichia coli.

    PubMed

    Liss, Michael A; Nakamura, Kristen K; Peterson, Ellena M

    2013-01-01

    A transrectal prostate biopsy is the most common procedure used to establish the diagnosis of prostate cancer. Prior to biopsy, patients are commonly given ciprofloxacin for prophylaxis. However, a complication of the procedure is infection with ciprofloxacin-resistant organisms, in particular resistant Escherichia coli. In order to identify patients carrying ciprofloxacin-resistant E. coli, so as to tailor their antibiotic prophylaxis, rectal swabs are screened using selective broth and/or solid medium. In our evaluation, we compared broth enrichment and direct plating techniques by using brain heart infusion broth and MacConkey agar containing 1 μg/ml or 10 μg/ml of ciprofloxacin. Of the 100 patients included in the study, 20 were colonized with ciprofloxacin-resistant organisms, 19 of which were E. coli. There was no significant difference (P > 0.1) between the culture methods or the ciprofloxacin concentrations in the medium when identifying patients with ciprofloxacin-resistant E. coli; however, broth enrichment using 1 μg/ml ciprofloxacin was the most sensitive at 100%, but it was the least specific. Direct plating of rectal swabs onto MacConkey agar containing 10 μg/ml of ciprofloxacin was 100% specific and missed only 1 positive specimen, with a sensitivity of 94.7%; this method was the most cost-effective. Therefore, direct plating of rectal swabs onto selective medium proved to be a sensitive and cost-effective approach in identifying patients colonized with ciprofloxacin-resistant E. coli.

  6. 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

  7. Tamoxifen induces permanent growth arrest through selective induction of apoptosis in growth plate chondrocytes in cultured rat metatarsal bones.

    PubMed

    Chagin, Andrei S; Karimian, Elham; Zaman, Farasat; Takigawa, Masaharu; Chrysis, Dionisios; Sävendahl, Lars

    2007-05-01

    Estrogen affects skeletal growth and promotes growth plate fusion in humans. High doses of estrogen have been used to limit growth in girls with predicted extreme tall stature; a treatment which has been associated with severe side effects. Selective estrogen receptor modulators (SERMs) could potentially be used as an alternative treatment. We chose to study the effects of Tamoxifen (Tam), a first generation SERM that has been used in the treatment of pubertal gynecomastia or McCune-Albright syndrome. Cultured fetal rat metatarsal bones were used to study the effects of Tam on longitudinal bone growth. In sectioned bones, chondrocyte apoptosis and proliferation were analyzed by TUNEL assay and BrdU incorporation, respectively. We also used a human chondrocytic cell line, HSC-2/8, to study the effects of Tam on apoptosis (FACS analysis and Cell Death detection ELISA) and caspase activation (caspase substrate cleavage and Western immunoblotting). Tam caused a dose-dependent growth retardation of cultured metatarsal bones. No catch-up growth was observed after Tam was removed from the culture medium. Detailed analysis of sectioned growth plate cartilage revealed increased apoptosis of chondrocytes within the resting and hypertrophic zones. HCS-2/8 cells also underwent apoptosis upon Tam treatment. Tam-induced apoptosis was caspase-dependent and completely abrogated by either caspase-8 or -9 inhibitors. A substrate assay revealed that caspase-8 is first activated followed by caspase-9 and -3. Finally, FasL secretion was stimulated by Tam and blocking of either FasL or Fas decreased Tam-induced apoptosis in chondrocytes. We here describe a novel mechanism of tamoxifen-induced apoptosis in chondrocytes, involving the activation of caspases and the FasL/Fas pathway, which diminishes the potential for bone growth.

  8. 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

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

    PubMed

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

    2003-01-01

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

  10. Secretion of albumin and induction of CYP1A2 and CYP3A4 in novel three-dimensional culture system for human hepatocytes using micro-space plate.

    PubMed

    Nishimura, Masuhiro; Hagi, Mieko; Ejiri, Yoko; Kishimoto, Sanae; Horie, Toru; Narimatsu, Shizuo; Naito, Shinsaku

    2010-01-01

    We evaluated a novel primary three-dimensional culture system for human hepatocytes using micro-space plates. The functional activity of human hepatocytes in primary culture was determined by measuring albumin secretion from hepatocytes to medium and measuring expression levels of albumin, CYP1A2 and CYP3A4 mRNA. Albumin secretion was higher in micro-space plates compared with traditional plates after 72 h of culture; the levels of albumin secretion from hepatocytes to medium in culture using micro-space plates after 96 h of culture were 2.7-fold higher than those in culture using traditional plates, and secretion of albumin in micro-space plate culture subsequently remained constant. Expression levels of albumin, CYP1A2 and CYP3A4 mRNA in the culture of hepatocytes were significantly higher using micro-space plates than using traditional plates. The inducibility of CYP1A2 and CYP3A4 mRNA after exposure to inducers in hepatocyte culture on micro-space plates was comparable to that in culture on traditional plates, while expression of CYP1A2 and CYP3A4 mRNA after exposure to inducers was higher on micro-space plates than on traditional plates. The present study demonstrates that a novel primary three-dimensional culture system of cryopreserved human hepatocytes using micro-space plates could be used for evaluating the induction of drug-metabolizing enzymes in humans. This in vitro method may thus be useful for screening the induction potency of new drug candidates.

  11. 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

  12. 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.

  13. Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits.

    PubMed

    Kanner, Sivan; Bisio, Marta; Cohen, Gilad; Goldin, Miri; Tedesco, Marieteresa; Hanein, Yael; Ben-Jacob, Eshel; Barzilai, Ari; Chiappalone, Michela; Bonifazi, Paolo

    2015-04-15

    The brain operates through the coordinated activation and the dynamic communication of neuronal assemblies. A major open question is how a vast repertoire of dynamical motifs, which underlie most diverse brain functions, can emerge out of a fixed topological and modular organization of brain circuits. Compared to in vivo studies of neuronal circuits which present intrinsic experimental difficulties, in vitro preparations offer a much larger possibility to manipulate and probe the structural, dynamical and chemical properties of experimental neuronal systems. This work describes an in vitro experimental methodology which allows growing of modular networks composed by spatially distinct, functionally interconnected neuronal assemblies. The protocol allows controlling the two-dimensional (2D) architecture of the neuronal network at different levels of topological complexity. A desired network patterning can be achieved both on regular cover slips and substrate embedded micro electrode arrays. Micromachined structures are embossed on a silicon wafer and used to create biocompatible polymeric stencils, which incorporate the negative features of the desired network architecture. The stencils are placed on the culturing substrates during the surface coating procedure with a molecular layer for promoting cellular adhesion. After removal of the stencils, neurons are plated and they spontaneously redirected to the coated areas. By decreasing the inter-compartment distance, it is possible to obtain either isolated or interconnected neuronal circuits. To promote cell survival, cells are co-cultured with a supporting neuronal network which is located at the periphery of the culture dish. Electrophysiological and optical recordings of the activity of modular networks obtained respectively by using substrate embedded micro electrode arrays and calcium imaging are presented. While each module shows spontaneous global synchronizations, the occurrence of inter-module synchronization

  14. 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.

  15. Comparison of the transcriptomic profile of hepatic human induced pluripotent stem like cells cultured in plates and in a 3D microscale dynamic environment.

    PubMed

    Leclerc, Eric; Kimura, Keiichi; Shinohara, Marie; Danoy, Mathieu; Le Gall, Morgane; Kido, Taketomo; Miyajima, Atsushi; Fujii, Teruo; Sakai, Yasuyuki

    2017-01-01

    We have compared the transcriptomic profiles of human induced pluripotent stem cells after their differentiation in hepatocytes like cells in plates and microfluidic biochips. The biochips provided a 3D and dynamic support during the cell differentiation when compared to the 2D static cultures in plates. The microarray have demonstrated the up regulation of important pathway related to liver development and maturation during the culture in biochips. Furthermore, the results of the transcriptomic profile, coupled with immunostaining, and RTqPCR analysis have shown typical biomarkers illustrating the presence of responders of biliary like cells, hepatocytes like cells, and endothelial like cells. However, the overall tissue still presented characteristic of immature and foetal patterns. Nevertheless, the biochip culture provided a specific micro-environment in which a complex multicellular differentiation toward liver could be oriented.

  16. 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.

  17. Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates.

    PubMed

    Shi, Handuo; Colavin, Alexandre; Lee, Timothy K; Huang, Kerwyn Casey

    2017-02-01

    Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.

  18. A 6 x 6 drop plate method for simultaneous colony counting and MPN enumeration of Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli.

    PubMed

    Chen, Chin Yi; Nace, Gary W; Irwin, Peter L

    2003-11-01

    A protocol was developed using 96-well plates and multichannel pipettes for serial dilutions, followed by drop plating on agar in a 6 x 6 format. This protocol permits simultaneous plating of six dilutions which greatly decreases the number of plates utilized thereby saving incubator space for organisms such as Campylobacter which require unique environmental conditions.

  19. Liquid chromatography/tandem mass spectrometric bioanalysis using normal-phase columns with aqueous/organic mobile phases - a novel approach of eliminating evaporation and reconstitution steps in 96-well SPE.

    PubMed

    Naidong, Weng; Shou, Wilson Z; Addison, Thomas; Maleki, Saber; Jiang, Xiangyu

    2002-01-01

    Bioanalytical methods using automated 96-well solid-phase extraction (SPE) and liquid chromatography with electrospray tandem mass spectrometry (LC/MS/MS) are widely used in the pharmaceutical industry. SPE methods typically require manual steps of drying of the eluates and reconstituting of the analytes with a suitable injection solvent possessing elution strength weaker than the mobile phase. In this study, we demonstrated a novel approach of eliminating these two steps in 96-well SPE by using normal-phase LC/MS/MS methods with low aqueous/high organic mobile phases, which consisted of 70-95% organic solvent, 5-30% water, and small amount of volatile acid or buffer. While the commonly used SPE elution solvents (i.e. acetonitrile and methanol) have stronger elution strength than a mobile phase on reversed-phase chromatography, they are weaker elution solvents than a mobile phase for normal-phase LC/MS/MS and therefore can be injected directly. Analytical methods for a range of polar pharmaceutical compounds, namely, omeprazole, metoprolol, fexofenadine, pseudoephedrine as well as rifampin and its metabolite 25-desacetyl-rifampin, in biological fluids, were developed and optimized based on the foregoing principles. As a result of the time saving, a batch of 96 samples could be processed in one hour. These bioanalytical LC/MS/MS methods were validated according to "Guidance for Industry - Bioanalytical Method Validation" recommended by the Food and Drug Administration (FDA) of the United States.

  20. 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.

  1. Effect of different culture systems and 3, 5, 3'-triiodothyronine/follicle-stimulating hormone on preantral follicle development in mice.

    PubMed

    Zhang, Cheng; Wang, Xiaoxia; Wang, Zhengpin; Niu, Wanbao; Zhu, Baochang; Xia, Guoliang

    2013-01-01

    The mechanical method to isolate preantral follicle has been reported for many years. However, the culture systems in vitro are still unstable. The aim of this study was to analyze the effect of the culture system of mice preantral follicles on the follicular development in vitro. The results showed that the 96-well plate system was the most effective method for mice follicle development in vitro (volume change: 51.71%; survival rate: 89%, at day 4). Follicle-stimulating hormone (FSH) and Thyroid hormone (TH) are important for normal follicular development and dysregulation of hormones are related with impaired follicular development. To determine the effect of hormone on preantral follicular development, we cultured follicle with hormones in the 96-well plate culture system and found that FSH significantly increased preantral follicular growth on day 4. The FSH-induced growth action was markedly enhanced by T₃ although T₃ was ineffective alone. We also demonstrated by QRT-PCR that T₃ significantly enhanced FSH-induced up-regulation of Xiap mRNA level. Meanwhile, Bad, cell death inducer, was markedly down-regulated by the combination of hormones. Moreover, QRT-PCR results were also consistent with protein regulation which detected by Western Blotting analysis. Taken together, the findings of the present study demonstrate that 96-well plate system is an effective method for preantral follicle development in vitro. Moreover, these results provide insights on the role of thyroid hormone in increasing FSH-induced preantral follicular development, which mediated by up-regulating Xiap and down-regulating Bad.

  2. 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.

  3. 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

  4. 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

  5. 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 (...

  6. Differential specificity of selective culture media for enumeration of pathogenic vibrios: advantages and limitations of multi-plating methods.

    PubMed

    Nigro, Olivia D; Steward, Grieg F

    2015-04-01

    Plating environmental samples on vibrio-selective chromogenic media is a commonly used technique that allows one to quickly estimate concentrations of putative vibrio pathogens or to isolate them for further study. Although this approach is convenient, its usefulness depends directly on how well the procedure selects against false positives. We tested whether a chromogenic medium, CHROMagar Vibrio (CaV), used alone (single-plating) or in combination (double-plating) with a traditional medium thiosulfate-citrate-bile-salts (TCBS), could improve the discrimination among three pathogenic vibrio species (Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus) and thereby decrease the number of false-positive colonies that must be screened by molecular methods. Assays were conducted on water samples from two estuarine environments (one subtropical, one tropical) in a variety of seasonal conditions. The results of the double-plating method were confirmed by PCR and 16S rRNA sequencing. Our data indicate that there is no significant difference in the false-positive rate between CaV and TCBS when using a single-plating technique, but determining color changes on the two media sequentially (double-plating) reduced the rate of false positive identification in most cases. The improvement achieved was about two-fold on average, but varied greatly (from 0- to 5-fold) and depended on the sampling time and location. The double-plating method was most effective for V. vulnificus in warm months, when overall V. vulnificus abundance is high (false positive rates as low as 2%, n=178). Similar results were obtained for V. cholerae (minimum false positive rate of 16%, n=146). In contrast, the false positive rate for V. parahaemolyticus was always high (minimum of 59%, n=109). Sequence analysis of false-positive isolates indicated that the majority of confounding isolates are from the Vibrionaceae family, however, members of distantly related bacterial groups were also able to

  7. Enumeration of starter cultures during yogurt production using Petrifilm AC plates associated with acidified MRS and M17 broths.

    PubMed

    Gonçalves, Marília M; Freitas, Rosangela; Nero, Luís A; Carvalho, Antônio F

    2009-05-01

    The efficiency of Petrifilm AC (3M Microbiology, St. Paul, MN, USA) associated with the broths M17 and de Mann-Rogosa-Sharpe (MRS) at pH 5.4 was evaluated to enumerate Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus during the yogurt production. Commercial and reference strains of these microorganisms were experimentally inoculated in nonfat milk and incubated at 42 degrees C for 4 h for yogurt production. At the moment of inoculation and after incubation, aliquots were collected, submitted to dilution using the broths M17 and MRS at pH 5.4, and plated for Strep. salivarius and Lb. bulgaricus enumeration according ISO 9232 and at Petrifilm AC plates, respectively. M17 plates were incubated at 42 degrees C, and MRS plates were incubated at 35 degrees C under anaerobiosis. After 48 h, the formed colonies were enumerated and the counts were compared by correlation and analysis of variance (P<0.05). In addition, colonies were randomly selected from all plates and characterized according to Gram staining and morphology. The obtained results indicated that Petrifilm AC plates associated to M17 and MRS at pH 5.4 can be considered as a suitable alternative for Strep. salivarius and Lb. bulgaricus enumeration during yogurt production, with slight interferences due to the acidity of MRS at the moment of inoculation, and due to the acidity of yogurt at the end of fermentation process. It was also observed that the MRS at pH 5.4 was not sufficiently selective for Lb. delbrueckii enumeration, despite it is indicated by the official protocol from ISO 9232.

  8. 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.

  9. Establishment of a fully automated microtiter plate-based system for suspension cell culture and its application for enhanced process optimization.

    PubMed

    Markert, Sven; Joeris, Klaus

    2017-01-01

    We developed an automated microtiter plate (MTP)-based system for suspension cell culture to meet the increased demands for miniaturized high throughput applications in biopharmaceutical process development. The generic system is based on off-the-shelf commercial laboratory automation equipment and is able to utilize MTPs of different configurations (6-24 wells per plate) in orbital shaken mode. The shaking conditions were optimized by Computational Fluid Dynamics simulations. The fully automated system handles plate transport, seeding and feeding of cells, daily sampling, and preparation of analytical assays. The integration of all required analytical instrumentation into the system enables a hands-off operation which prevents bottlenecks in sample processing. The modular set-up makes the system flexible and adaptable for a continuous extension of analytical parameters and add-on components. The system proved suitable as screening tool for process development by verifying the comparability of results for the MTP-based system and bioreactors regarding profiles of viable cell density, lactate, and product concentration of CHO cell lines. These studies confirmed that 6 well MTPs as well as 24 deepwell MTPs were predictive for a scale up to a 1000 L stirred tank reactor (scale factor 1:200,000). Applying the established cell culture system for automated media blend screening in late stage development, a 22% increase in product yield was achieved in comparison to the reference process. The predicted product increase was subsequently confirmed in 2 L bioreactors. Thus, we demonstrated the feasibility of the automated MTP-based cell culture system for enhanced screening and optimization applications in process development and identified further application areas such as process robustness. The system offers a great potential to accelerate time-to-market for new biopharmaceuticals. Biotechnol. Bioeng. 2017;114: 113-121. © 2016 Wiley Periodicals, Inc.

  10. Chikungunya virus isolation using simplified cell culture technique in Mauritius.

    PubMed

    Pyndiah, M N; Pursem, V; Meetoo, G; Daby, S; Ramuth, V; Bhinkah, P; Chuttoo, R; Paratian, U

    2012-03-01

    During the chikungunya outbreak of 2005 - 2006, the only laboratory facilities available in Mauritius were virus isolation in cell culture tubes and serology. The laboratory was submerged with large numbers of blood samples. Comparative isolation was made in human embryonic lung (HEL) and VERO cells grown in 96-well plate. Culture on HEL cells was found to be more sensitive and presence of cytopathic effect (CPE) was observed earlier than in VERO cells. Out of the 18 300 blood samples inoculated on HEL, 11 165 were positive. This virus isolation method was of great help for the surveillance and control of the vectors. In cases of an outbreak a cheap, rapid and simple method of isolating chikungunya virus is described.

  11. Toxicity of organotin compounds in primary cultures of rat cortical astrocytes.

    PubMed

    Röhl, C; Gülden, M; Seibert, H

    2001-01-01

    The neurotoxic organotin compounds trimethyl (TMT) and triethyltin (TET) are known to induce astrogliosis in vivo, which is indicated by an increased synthesis of glial fibrillary acidic protein (GFAP) in astrocytes. In contrast, tributyltin (TBT) does not induce astrogliosis. The aim of this study was to investigate whether trialkyltin derivatives can induce an increased GFAP synthesis in astrocyte cultures in the absence of neurons and whether differences between the action of TMT, TET, and TBT can be detected. Primary cultures of rat cortical astrocytes from 2-day-old rats were grown in 96-well plates until confluency and then exposed to various concentrations of TMT, TET, and TBT for 40 h. Effects on basal cell functions were measured by colorimetric determination of cell protein contents and by assessment of viability by means of the MTT assay. An indirect sandwich ELISA for 96-well plates was used for quantitative measurements of the GFAP content of the cells. All three compounds induced a concentration-dependent cytotoxicity indicated by parallel decreases of protein contents and MTT reduction. Half-maximum cytotoxic concentrations were 3 micromol/L (TBT), 30 micromol/L (TET), and 800 micromol/L (TMT). Cellular GFAP contents were reduced in parallel to cytotoxic action but no increase in GFAP expression at subcytotoxic concentrations could be observed. Thus, the astrocytes were not able to respond to TMT or TET exposure by an increased synthesis of GFAP in the absence of neuronal signals.

  12. A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates.

    PubMed

    Mochida, Seiya; Tsuzuki, Satoshi; Inouye, Kuniyo; Fushiki, Tohru

    2014-05-01

    Matriptase is an epithelial-derived type-II transmembrane serine protease. This protease is expressed prominently in the villus tip of small-intestinal epithelia at which senescent cells undergo shedding and/or apoptosis. The basement membrane of epithelial cells, including small-intestinal epithelial cells, contains extracellular matrix (ECM) proteins such as fibronectin and laminin. We found previously that high concentrations of a recombinant matriptase catalytic domain (r-MatCD) (e.g. 1 μM) caused an increased detachment of and increases in the activity of apoptotic effector caspase-3 in a rat small-intestinal epithelial IEC-6 line cultured on laminin-coated plates and proposed that at sites with its high level of expression, matriptase contributes to promoting shedding and/or detachment-induced death of epithelial cells through a mechanism mediating loss of cell-ECM adhesion. In this study, we found that even without increasing cell detachment, a high concentration of r-MatCD causes an increase in caspase-3 activity in IEC-6 cells cultured on fibronectin-coated plates, suggesting that the recombinant matriptase can cause apoptosis by a mechanism unrelated to cell detachment. Also, r-MatCD-treated IEC-6 cells on fibronectin were found to display spindle-like morphological changes. We suggest that r-MatCD causes apoptosis of IEC-6 on fibronectin by a mechanism involving the disruption of cell integrity.

  13. 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

  14. Validation and application of a liquid chromatography-tandem mass spectrometric method for the determination of GDC-0834 and its metabolite in human plasma using semi-automated 96-well protein precipitation.

    PubMed

    Shin, Young G; Jones, Steve A; Murakami, Stan C; Liu, Lichuan; Wong, Harvey; Buonarati, Michael H; Hop, Cornelis E C A

    2012-11-01

    A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the determination of GDC-0834 and its amide hydrolysis metabolite (M1) in human plasma to support clinical development. The method consisted of semi-automated 96-well protein precipitation extraction for sample preparation and LC-MS/MS analysis in positive ion mode using TurboIonSpray® for analysis. D6-GDC-0834 and D6-M1 metabolite were used as internal standards. A linear regression (weighted 1/concentration(2) ) was used to fit calibration curves over the concentration range of 1 - 500 ng/mL for both GDC-0834 and M1 metabolite. The accuracy (percentage bias) at the lower limit of quantitation (LLOQ) was 5.20 and 0.100% for GDC-0834 and M1 metabolite, respectively. The precision (CV) for samples at the LLOQ was 3.13-8.84 and 5.20-8.93% for GDC-0834 and M1 metabolite, respectively. For quality control samples at 3, 200 and 400 ng/mL, the between-run CV was ≤ 7.38% for GDC-0834 and ≤ 8.20% for M1 metabolite. Between run percentage bias ranged from -2.76 to 6.98% for GDC-0834 and from -6.73 to 2.21% for M1 metabolite. GDC-0834 and M1 metabolite were stable in human plasma for 31 days at -20 and -70°C. This method was successfully applied to support a GDC-0834 human pharmacokinetic-based study.

  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. High-throughput, semi-automated determination of a cyclooxygenase II inhibitor in human plasma and urine using solid-phase extraction in the 96-well format and high-performance liquid chromatography with post-column photochemical derivatization-fluorescence detection.

    PubMed

    Matthews, C Z; Woolf, E J; Lin, L; Fang, W; Hsieh, J; Ha, S; Simpson, R; Matuszewski, B K

    2001-02-25

    Compound I, 5-chloro-3-(4-methanesulfonylphenyl)-6'-methyl-[2,3']bipyridinyl, has been found to be a specific inhibitor of the enzyme cyclooxygenase II (COX II). The anti-inflammatory properties of this compound are currently being investigated. HPLC assays for the determination of this analyte in human plasma and human urine have been developed. Isolation of I and the internal standard (II) was achieved by solid-phase extraction (SPE) in the 96-well format. A C8 SPE plate was used for the extraction of the drug from human plasma (recovery >90%) while a mixed-mode (C8/Cation) SPE plate was used to isolate the analytes from human urine (recovery approximately 71%). The analyte and internal standard were chromatographed on a Keystone Scientific Prism-RP guard column (20 x 4.6 mm) connected to a Prism-RP analytical column (150 x 4.6 mm), using a mobile phase consisting of 45% acetonitrile in 10 mM acetate buffer (pH = 4); the analytes eluted at retention times of 5.2 and 6.9 min for I and II, respectively. Compounds I and II were found to form highly fluorescent products after exposure to UV light (254 nm). Thus, the analytes were detected by fluorescence (lambda(ex) = 260 nm, lambda(em) =375 nm) following post-column photochemical derivatization. Eight point calibration curves over the concentration range of 5-500 ng/ml for human plasma and human urine yielded a linear response (R2>0.99) when a 1/y weighted linear regression model was employed. Based on the replicate analyses (n = 5) of spiked standards, the within-day precision for both assays was better than 7% C.V. at all points on the calibration curve; within-day accuracy was within 5% of nominal at all standard concentrations. The between-run precision and accuracy of the assays, as calculated from the results of the analysis of quality control samples, was better than 8% C.V. and within 8% of nominal. I was found to be stable in human plasma and urine for at least 8 and 2 months, respectively. In addition, the

  17. Adaptive Focused Acoustics (AFA) Improves the Performance of Microtiter Plate ELISAs.

    PubMed

    Green, David J; Rudd, Edwin A; Laugharn, James A

    2014-08-01

    We investigated the use of Adaptive Focused Acoustics (AFA) technology to improve the performance of microtiter plate enzyme-linked immunosorbent assays (ELISAs). Experiments were performed with commercially available AFA instrumentation and off-the-shelf 96-well microtiter plate sandwich ELISAs. AFA was applied over a range of acoustic energies, temperatures, and durations to the antigen/antibody binding step of an ELISA for measuring HIV-1 p24 in tissue culture samples. AFA-mediated antigen/antibody binding was enhanced up to 2-fold over passive binding at comparable temperatures and was superior or comparable at low temperature (8-10 °C) to passive binding at 37 °C. Lower nonspecific binding (NSB), lower inter- and intra-assay coefficients of variation (CVs), higher Z' factors, and lower limits of detection (LODs) were measured in AFA-mediated assays compared with conventional passive binding. In a more limited study, AFA enhancement of antigen/antibody binding and lower NSB was measured in an ELISA for measuring IGFBP-3 in human plasma. We conclude from this study that application of AFA to antigen/antibody binding steps in microtiter plate ELISAs can enhance key assay performance parameters, particularly Z' factors and LODs. These features render AFA-mediated binding assays potentially more useful in applications such as high-throughput screening and in vitro diagnostics than assays processed with conventional passive antigen/antibody binding steps.

  18. 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.

  19. Induction of osteogenic differentiation of stem cells via a lyophilized microRNA reverse transfection formulation on a tissue culture plate.

    PubMed

    Wu, Kaimin; Xu, Jie; Liu, Mengyuan; Song, Wen; Yan, Jun; Gao, Shan; Zhao, Lingzhou; Zhang, Yumei

    2013-01-01

    MicroRNA (miRNA) regulation is a novel approach to manipulating the fate of mesenchymal stem cells, but an easy, safe, and highly efficient method of transfection is required. In this study, we developed an miRNA reverse transfection formulation by lyophilizing Lipofectamine 2000-miRNA lipoplexes on a tissue culture plate. The lipoplexes can be immobilized on a tissue culture plate with an intact pseudospherical structure and lyophilization without any lyoprotectant. In this study, reverse transfection resulted in highly efficient cellular uptake of miRNA and enabled significant manipulation of the intracellular target miRNA level. Reverse transfection formulations containing Lipofectamine 2000 1 μL per well generated much higher transfection efficiency without obvious cytotoxicity compared with conventional and other transfection methods. Further, the transfection efficiency of the reverse transfection formulations did not deteriorate during 90 days of storage at 4°C and -20°C. We then assessed the efficiency of the miRNA reverse transfection formulation in promoting osteogenic differentiation of mesenchymal stem cells. We found that transfection with anti-miR-138 and miR-148b was efficient for enhancing osteogenic differentiation, as indicated by enhanced osteogenesis-related gene expression, amount of alkaline phosphatase present, production of collagen, and matrix mineralization. Overall, the miRNA reverse transfection formulation developed in this study is a promising approach for miRNA transfection which can control stem cell fate and is suitable for loading miRNAs onto various biomaterials.

  20. Mixing at the microscale: Power input in shaken microtiter plates.

    PubMed

    Dürauer, Astrid; Hobiger, Stefanie; Walther, Cornelia; Jungbauer, Alois

    2016-12-01

    Power input and local energy dissipation are crucial parameters for the engineering characterization of mixing and fluid dynamics at the microscale. Since hydrodynamic stress is solely dependent on the maximum power input, we adapted the clay/polymer method to obtain flock destruction kinetics in six-, 24-, and 96-well microtiter plates on orbital shakers. We also determined the specific power input using calorimetry and found that the power input is at the same order of magnitude for the six- and 96-well plates and the laboratory-scale stirred tank reactor, with 40 to 90 W/m(3) (Re' = 180 to 440), 40 to 140 W/m3 (Re' = 320 to 640), and 30 to 50 W/m(3) (Re = 4000 to 8500), respectively. All of these values are significantly below 450 to 2100 W/m(3) determined for the pilot-scale reactor. The hydrodynamic stress differs significantly between the different formats of MTPs, as the 96-well plates showed very low shear stress on the shaker with a shaking amplitude of 3 mm. Thus, the transfer of mixing conditions from the microtiter plate to small-scale and pilot-scale reactors must be undertaken with care. Our findings, especially the power input determined by the calorimetric method, show that the hydrodynamic conditions in laboratory- and pilot-scale reactors cannot be reached.

  1. Comparison study of streptavidin-coated microtitration plates.

    PubMed

    Välimaa, Lasse; Laurikainen, Katja

    2006-01-20

    Streptavidin (SAv)-coated 96-well microtitration plates from commercial suppliers (six plates) and our laboratory (two plates) were tested with respect to their binding capacity for small and large molecules, leaching, well-to-well variation and immunoassay performance. The binding capacities for europium-labelled biotin (Eu-biotin) varied from 4.4 to above 150 pmol/well depending on the plate, and for biotinylated monoclonal antibody from 1.2 to 6.4 pmol/well (190-1030 ng). Incubation for 1 h in regular immunoassay buffer resulted in leaching of 0.6-76 ng of SAv from unwashed wells and 0.5-60 ng from pre-washed wells. The desorbed quantity represented up to 3.4% of the maximal Eu-biotin binding capacity. Coating-related variation of 96 wells (CV% values) ranged from 1.2% to 8.0% when tested with respect to maximum binding capacity. Immunofluorometric assay for TSH as well as enzyme immunoassays for CA125 and PSA revealed immunoassay-related performance of the plates, regarding signal levels, variation and non-specific binding characteristics. Non-specific binding and variation tended to increase in the highest capacity plates, whereas some low capacity plates fulfilled the assay requirements optimally. Good performance with respect to capacity, coating homogeneity or leaching was not by definition reflected in the performance of two-site heterogeneous immunoassays.

  2. 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.

  3. Microtiter plate transformation using the LiAc/SS carrier DNA/PEG method.

    PubMed

    Gietz, R Daniel; Schiestl, Robert H

    2007-01-01

    Here, we describe a protocol that has been adapted for the transformation of yeast cells in 96-well microtiter plates. This protocol can be tailored for multiple applications and is suitable for high-throughput applications. It can be completed in 2-3 h, once the yeast cells have been grown depending on the heat shock used.

  4. 3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates.

    PubMed

    Vrij, E J; Espinoza, S; Heilig, M; Kolew, A; Schneider, M; van Blitterswijk, C A; Truckenmüller, R K; Rivron, N C

    2016-02-21

    3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.

  5. Imaging and quantitative analysis of tritium-labelled cells in lymphocyte proliferation assays using microchannel plate detectors originally developed for X-ray astronomy.

    PubMed

    Lees, J E; Hales, J M

    2001-01-01

    Microchannel plate detectors have been used in many astronomical X-ray telescopes. Recently we have begun to use similar detectors to image electron emission from radiolabelled biological assays. Here we show how a microchannel plate (MCP) detector can be used to image tritium uptake in T lymphocyte proliferation assays. Quantitative analysis using the MCP detector has the same sensitivity and speed as conventional liquid scintillation counter (LSC) analysis whilst obviating the need for scintillation fluid. In addition the system permits the imaging of whole plate harvests from a range of plate sizes. Here we present data obtained with 96-well plates and Terasaki plates.

  6. A colorimetric method for the measurement of platelet adhesion in microtiter plates.

    PubMed

    Bellavite, P; Andrioli, G; Guzzo, P; Arigliano, P; Chirumbolo, S; Manzato, F; Santonastaso, C

    1994-02-01

    A procedure for the determination of the adhesion of human platelets to protein-coated culture microwells was developed. The number of platelets was quantitated by measuring the activity of acid phosphatase, a platelet enzyme whose activity is stable independently of platelet stimulation and is not released. Isolated and washed platelets were incubated in 96-well microtiter plates with flat-bottom wells that had been precoated with various compounds, including collagen, fibrinogen, human plasma, and human albumin. At the end of incubation (optimal time: 40-60 min), nonadherent platelets were washed out, adherent platelets were solubilized with Triton X-100, and the acid phosphatase activity was measured by using the substrate p-nitrophenyl phosphate. The p-nitrophenol produced was measured with a microplate reader at 405 nm and the percentage of adhesion was calculated with reference to known platelet standards. ADP and thrombin stimulated platelet adhesion in a dose-dependent manner to fibrinogen and human plasma, but not to human albumin. Platelets adhered to collagen even in the absence of stimulants. Simultaneous evaluation of adhesion and aggregation demonstrated that with ADP as stimulus, but not with thrombin, the two platelet responses were dissociated. Microscopic examination of culture wells showed that most of platelets adhered as single cells and not as aggregates. The sensitivity of this method allowed the assay of platelet adhesion by using only 2.5 x 10(5) platelets/well.

  7. Robotic platform for parallelized cultivation and monitoring of microbial growth parameters in microwell plates.

    PubMed

    Knepper, Andreas; Heiser, Michael; Glauche, Florian; Neubauer, Peter

    2014-12-01

    The enormous variation possibilities of bioprocesses challenge process development to fix a commercial process with respect to costs and time. Although some cultivation systems and some devices for unit operations combine the latest technology on miniaturization, parallelization, and sensing, the degree of automation in upstream and downstream bioprocess development is still limited to single steps. We aim to face this challenge by an interdisciplinary approach to significantly shorten development times and costs. As a first step, we scaled down analytical assays to the microliter scale and created automated procedures for starting the cultivation and monitoring the optical density (OD), pH, concentrations of glucose and acetate in the culture medium, and product formation in fed-batch cultures in the 96-well format. Then, the separate measurements of pH, OD, and concentrations of acetate and glucose were combined to one method. This method enables automated process monitoring at dedicated intervals (e.g., also during the night). By this approach, we managed to increase the information content of cultivations in 96-microwell plates, thus turning them into a suitable tool for high-throughput bioprocess development. Here, we present the flowcharts as well as cultivation data of our automation approach.

  8. A cost-effective plate-based sample preparation for antibody N-glycan analysis.

    PubMed

    Burnina, Irina; Hoyt, Erik; Lynaugh, Heather; Li, Huijuan; Gong, Bing

    2013-09-13

    During early cell line and process development of therapeutic antibodies, a cost-effective high-throughput approach to characterize the N-linked glycans is highly desired given that a large number of samples need to be analyzed. Using commercially available, low cost 96-well plates, we developed a practical procedure to prepare fluorescently labeled N-linked glycans for both qualitative and quantitative analysis by mass spectrometry (MS) and ultrahigh performance liquid chromatography (UPLC). Antibody samples were continuously denatured, reduced, and deglycosylated in a single 96-well hydrophobic membrane filter plate. Subsequently, released glycans were fluorescently labeled in a collection plate, and cleaned-up using a hydrophilic membrane filter plate. Carried out entirely in ready-to-use 96-well plates with simple buffer systems, this procedure requires less than 90min to finish. We applied the optimized procedure to examine the N-linked glycosylation of trastuzumab and were able to quantify ten major N-linked glycans. The results from different amounts of starting materials (10-200μg) were highly similar and showed the robustness of this procedure. Compared to other methods, this new procedure is simple to implement, economically more affordable, and could be very valuable for early screenings of antibody development.

  9. 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.

  10. A novel method for toxicology: in vitro culture system of a rat preantral follicle.

    PubMed

    Wan Xuying; Zhu Jiangbo; Zhu Yuping; Xili, Ma; Liu Zhen; Wang Fei; Xu Guifeng; Zhang Tianbao

    2011-08-01

    Preantral follicle in vitro culture systems have been successfully or nearly successfully established for sheep, pig and mouse, and applied on follicle development and regulation research on reproductive biology and physiology. However, there have been few studies concerning rat preantral follicle in vitro development. The objective is to establish an in vitro culture system for rat preantral follicles which can be used for reproductive biology and toxicology research. Rat preantral follicles are mechanically separated, cultured in vitro in single follicle mode for continuous 12 days using 96-well plates, and then administrated ovulation induction. The observation on follicle development, hormone level, and ovum formation are recorded and assessed. Taking in vivo growth and in vitro maturation of oocytes group as control group, in vitro growth and maturation of oocytes group is assessed to see whether this in vitro culture method is successful. The conditions for rat follicle culture are determined based on the mouse pre-antral follicle culture. The in vitro culture system for rat preantral follicles established in this study is feasible and successful, and can serve as model for reproductive biology and toxicology research.

  11. 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…

  12. Parallel electromembrane extraction in the 96-well format.

    PubMed

    Eibak, Lars Erik Eng; Rasmussen, Knut Einar; Oiestad, Elisabeth Leere; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2014-05-30

    The repeatability and extraction recoveries of parallel electromembrane extraction (Pa-EME) was thoroughly investigated in the present project. Amitriptyline, fluoxetine, and haloperidol were isolated from eight samples of pure water, undiluted human plasma, and undiluted human urine, respectively; in total 24 samples were processed in parallel. The repeatability was found to be independent of the different sample matrices (pure water samples, human plasma, and water) processed in parallel, although the respective samples contained different matrix components. In another experiment seven of the 24 wells were perforated. Even though the perforation caused the total current level in the Pa-EME setup to increase, the intact circuits were unaffected by the collapse in seven of the circuits. In another approach, exhaustive extraction of amitriptyline, fluoxetine, and haloperidol was demonstrated from pure water samples. Amitriptyline and haloperidol were also isolated exhaustively from undiluted human plasma samples; the extraction recovery of fluoxetine from undiluted human plasma was 81%. Finally, the sample throughput was increased with the Pa-EME configuration. The extraction recoveries were investigated by processing 1, 8, 68, or 96 samples in parallel in 10min; neither the extraction recoveries nor the repeatability was affected by the total numbers of samples. Eventually, the Pa-EME was combined with ultra performance liquid chromatography (UPLC) to combine high-throughput sample preparation with high-throughput analytical instrumentation. The aim of the present investigation was to demonstrate the potential of electromembrane extraction as a high throughput sample preparation platform; and hopefully to increase the interest for EME in the bioanalytical field to solve exisiting and novel analytical challenges.

  13. Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader.

    PubMed

    Pick, E; Mizel, D

    1981-01-01

    Two simple semiautomated microassays for the measurement of superoxide (O-2) and hydrogen peroxide (H2O2) production by cultured macrophages (MPs) are described. The measurement of O-2 is based on the reduction of ferricytochrome c as assayed by the increase in its absorbance at 550 nm. Quantitation of H2O2 is based on the horseradish peroxidase (HRPO)-dependent oxidation of phenol red which is assayed by its increased absorbance at 600 nm. MPs are cultured in monolayers in 96-well flat-bottom tissue culture plates and covered with 100 mul amounts per well of either a ferricytochrome c solution containing phenol red and HRPO. Following the addition of an agent eliciting an oxidative burst (OB) and incubation of the plates at 37 degrees C for various time intervals, the changes in the absorbance of ferricytochrome c and phenol red, respectively, are measured directly in the wells of the tissue culture plates with the cells in situ, by using an automatic 8-channel photometer which reads absorbances vertically through individual wells. This instrument, which was originally designed for reading enzyme immunoassays in microtitration plates, can be easily adapted for use in the above test, when fitted with interference filters with wave lengths of 550 nm (for the assay of O-2) and 600 nm (for the assay of H2O2). The principal advantages of this techniques are: the ability to perform the assays directly in the culture plates with cells in situ; the small amounts of cells and reagents needed; its sensitivity and reproducibility; the ease with which kinetic experiments can be done; the large number of samples which can be tested in parallel, and especially the speed and convenience offered by the automated reading and printout of absorbance values.

  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. 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.

  16. Aseptic laboratory techniques: plating methods.

    PubMed

    Sanders, Erin R

    2012-05-11

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

  17. 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

  18. Plate electronics

    NASA Astrophysics Data System (ADS)

    Carlowicz, Michael

    Using a Cray T3D supercomputer and a simple assumption about the physical character of Earth's mantle, a pair of researchers from the University of California at Berkeley have built a computer model that may help explain why the planet's tectonic plates look the way they do.In creating a three-dimensional numerical simulation of convection in the Earth's interior, UC researchers Hans-Peter Bunge and Mark Richards simplified their model to account for just one major physical effect: that the viscosity of the mantle increases with depth. Reviewing some recent—but not yet widely accepted—seismic data, Bunge and Richards assumed for the sake of the model that the viscosity of the mantle increases by a factor of 30 from the lithosphere to the core-mantle boundary. Relying on that assumption, the pair ran the model for nearly three weeks on a supercomputer at Los Alamos National Laboratory and found that the simulation produced an effect similar to what we see on the surface of Earth. The model produced a surface paralleling the actual width of plates and the geometry of the plate boundaries.

  19. pH measurement and a rational and practical pH control strategy for high throughput cell culture system.

    PubMed

    Zhou, Haiying; Purdie, Jennifer; Wang, Tongtong; Ouyang, Anli

    2010-01-01

    The number of therapeutic proteins produced by cell culture in the pharmaceutical industry continues to increase. During the early stages of manufacturing process development, hundreds of clones and various cell culture conditions are evaluated to develop a robust process to identify and select cell lines with high productivity. It is highly desirable to establish a high throughput system to accelerate process development and reduce cost. Multiwell plates and shake flasks are widely used in the industry as the scale down model for large-scale bioreactors. However, one of the limitations of these two systems is the inability to measure and control pH in a high throughput manner. As pH is an important process parameter for cell culture, this could limit the applications of these scale down model vessels. An economical, rapid, and robust pH measurement method was developed at Eli Lilly and Company by employing SNARF-4F 5-(-and 6)-carboxylic acid. The method demonstrated the ability to measure the pH values of cell culture samples in a high throughput manner. Based upon the chemical equilibrium of CO(2), HCO(3)(-), and the buffer system, i.e., HEPES, we established a mathematical model to regulate pH in multiwell plates and shake flasks. The model calculates the required %CO(2) from the incubator and the amount of sodium bicarbonate to be added to adjust pH to a preset value. The model was validated by experimental data, and pH was accurately regulated by this method. The feasibility of studying the pH effect on cell culture in 96-well plates and shake flasks was also demonstrated in this study. This work shed light on mini-bioreactor scale down model construction and paved the way for cell culture process development to improve productivity or product quality using high throughput systems.

  20. 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.

  1. Microenvironment Induced Spheroid to Sheeting Transition of Immortalized Human Keratinocytes (HaCaT) Cultured in Microbubbles Formed in Polydimethylsiloxane

    PubMed Central

    Chandrasekaran, Siddarth; Giang, Ut-Binh; King, Michael R.; DeLouise, Lisa A

    2011-01-01

    The in vivo cellular microenvironment is regulated by a complex interplay of soluble factors and signaling molecules secreted by cells and it plays a critical role in the growth and development of normal and diseased tissues. In vitro systems that can recapitulate the microenvironment at the cellular level are needed to investigate the influence of autocrine signaling and extracellular matrix effects on tissue homeostasis, regeneration, and disease development and progression. In this study we report the use of microbubble technology as a means to culture cells in a controlled microenvironment in which cells can influence their function through autocrine signaling. Microbubbles (MB) are small spherical cavities about 100–300 µm in diameter formed in hydrophobic polymer polydimethylsiloxane (PDMS) with ~60–100 µm circular openings and aspect ratio ~3.5. We demonstrate that the unique architecture of the microbubble compartment is advantaged for cell culture using HaCaT cells, an immortalized keratinocyte cell line. We observe that HaCaT cells, seeded in microbubbles (15–20 cells / MB) and cultured under standard conditions, adopt a compact 3-D spheroidal morphology. Within 2–3 days, the cells transition to a sheeting morphology. Through experimentation and simulation we show that this transition in morphology is due to the unique architecture of the microbubble compartment which enables cells to condition their local microenvironment. The small media volume per cell and the development of shallow concentration gradients allow factors secreted by the cells to rise to bioactive levels. The kinetics of the morphology transition depends on the number of cells seeded per microbubble; higher cell seeding induces a more rapid transition. HaCaT cells seeded onto PDMS cured in 96-well plates also form compact spheroids but they do not transition to a sheeting morphology even after several weeks of culture. The importance of soluble factor accumulation in driving

  2. Biodegradation of naphthenic acid surrogates by axenic cultures.

    PubMed

    Yue, Siqing; Ramsay, Bruce A; Ramsay, Juliana A

    2015-07-01

    This is the first study to report that bacteria from the genera Ochrobactrum, Brevundimonas and Bacillus can be isolated by growth on naphthenic acids (NAs) extracted from oil sands process water (OSPW). These pure cultures were screened for their ability to use a range of aliphatic, cyclic and aromatic NA surrogates in 96-well microtiter plates using water-soluble tetrazolium redox dyes (Biolog Redox Dye H) as the indicator of metabolic activity. Of the three cultures, Ochrobactrum showed most metabolic activity on the widest range of NA surrogates. Brevundomonas and especially Ochrobactrum had higher metabolic activity on polycyclic aromatic compounds than other classes of NA surrogates. Bacillus also oxidized a wide range of NA surrogates but not as well as Ochrobactrum. Using this method to characterize NA utilisation, one can identify which NAs or NA classes in OSPW are more readily degraded. Since aromatic NAs have been shown to have an estrogenic effect and polycyclic monoaromatic compounds have been suggested to pose the greatest environmental threat among the NAs, these bacterial genera may play an important role in detoxification of OSPW. Furthermore, this study demonstrates that bacteria belonging to the genera Ochrobactrum and Bacillus can also degrade surrogates of tricyclic NAs.

  3. 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.

  4. Prolonged cultures of unstimulated human neutrophils lead to the apparition and persistence of rest-in-plate structures (RIPs) recognized by professional phagocytes in vitro and in vivo.

    PubMed

    Vallières, Francis; Simard, Jean-Christophe; Stafford-Richard, Théo; Girard, Denis

    2015-12-01

    Polymorphonuclear neutrophil cells (PMNs) are known to spontaneously undergo apoptosis and then eliminated by professional phagocytes to prevent inflammation, a process called efferocytosis. However, when efferocytosis is impaired, PMNs will fall into secondary necrosis. Whether this state can persist for a certain period of time is unclear, since most of the studies investigating secondary necrosis are performed within 24h following induction by a proapoptotic agent. In this study, freshly isolated human PMNs were incubated without addition of exogenous agents in order to force them to undergo apoptosis and then secondary necrosis, an ideal experimental condition to study the behavior of secondary necrotic PMNs in absence of efferocytosis. By monitoring PMN cell morphology over time, we observed that an increasing proportion of cells harbored a ghost-like phenotype. Because these cellular remnants persist in plates for several days, we introduce here the terminology RIPs for 'rest-in-plate' structure. Heating of freshly isolated PMNs for 5min did not lead to the apparition of RIPs over time. In vivo administration of 7-days old RIPs in the murine air pouch model induced a slight inflammation resorbed within 24h. PKH26-stained RIPs were found to be ingested by professional phagocytes in vitro and in vivo in the murine air pouch and peritonitis models. Therefore, aged-PMNs have the potential to become RIPs in absence of efficient efferocytosis. Fortunately RIPs are recognized by professional phagocytes and, therefore, the concept of resolution of inflammation based on elimination of apoptotic and secondary necrotic PMNs could also be applied to RIPs.

  5. Superoxide dismutases from the oyster parasite Perkinsus marinus: purification, biochemical characterization, and development of a plate microassay for activity.

    PubMed

    Ahmed, Hafiz; Schott, Eric J; Gauthier, Julie D; Vasta, Gerardo R

    2003-07-01

    We have isolated and biochemically characterized superoxide dismutase (SOD) activity in cell extracts of clonally cultured Perkinsus marinus, a facultative intracellular parasite of the Eastern oyster, Crassostrea virginica. In order to assess the SOD activity throughout the purification, we developed and optimized a 96-well-plate microassay based on the inhibition of pyrogallol oxidation. The assay was also adapted to identify SOD activity type (Cu/Zn-, Mn-, or FeSOD), even in mixtures of more than one type of SOD. All SOD activity detected in the cell extracts was of the FeSOD type. Most of the SOD activity in P. marinus trophozoites resides in a major component of subunit molecular weight 24 kDa. The protein was purified by affinity chromatography on an anti-SOD antibody-Sepharose column. Amino-terminal peptide sequence of the affinity-purified protein corresponds to the predicted product of the PmSOD1 gene and indicates that amino-terminal processing has taken place. The results are discussed in the context of processing of mitochondrially targeted SODs.

  6. Corrugated cover plate for flat plate collector

    DOEpatents

    Hollands, K. G. Terry; Sibbitt, Bruce

    1978-01-01

    A flat plate radiant energy collector is providing having a transparent cover. The cover has a V-corrugated shape which reduces the amount of energy reflected by the cover away from the flat plate absorber of the collector.

  7. 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.

  8. Growth Plate Fractures

    MedlinePlus

    .org Growth Plate Fractures Page ( 1 ) The bones of children and adults share many of the same risks for injury. But because they ... to a unique injury called a growth plate fracture. Growth plates are areas of cartilage located near ...

  9. 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

  10. Simplification of aggregate culture of human mesenchymal stem cells as a chondrogenic screening assay.

    PubMed

    Welter, Jean F; Solchaga, Luis A; Penick, Kitsie J

    2007-06-01

    Aggregate culture provides a three-dimensional (3-D) environment for differentiating or differentiated cells; it is particularly useful to study in vitro chondrogenesis and cartilage biology. We have recently ported this method from a conical tube-based format to a 96-well plate format for the study of mesenchymal stem cell (MSC) chondrogenesis. The microplate format has greatly reduced the workload and materials cost, while maintaining reproducible chondrogenic differentiation. A long-term goal is to fully automate aggregate culture--this requires critically identifying all the indispensable steps of the protocol. Robotic laboratory equipment for manipulating microplate assays are commercially available; however centrifugation steps are difficult to implement automatically. We, therefore, tested whether the centrifugation step can be eliminated, thus significantly streamlining the assay workflow. By comparing aggregates prepared from human bone marrow-derived MSCs (hMSCs) that were formed either through centrifugation or through free sedimentation, we found that both methods produce aggregates with similar formation kinetics, and that there was no perceptible difference in the timing of the appearance of markers of chondrogenesis. Thus, it appears safe to eliminate the centrifugation step from the aggregate culture protocol. This results in significant time and effort savings and paves the way for future full automation of the aggregate assay.

  11. 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.

  12. 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

    2015-08-04

    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.

  13. 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

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

    NASA Astrophysics Data System (ADS)

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

    2016-09-01

    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.

  15. Microtiter micromass cultures of limb-bud mesenchymal cells.

    PubMed

    Paulsen, D F; Solursh, M

    1988-02-01

    A method is described for growing high-density micromass cultures of chick and mouse limb mesenchyme cells in 96-well microtiter plates (microT microM cultures). Rapid quantitative estimates of chondrogenic expression were obtained by automated spectrophotometric analysis of Alcian-blue-stained cartilage matrix extracts performed in the wells in which the cells had been grown. Quantitative estimates of myogenic expression were obtained similarly using anti-sarcomere myosin monoclonal antibody and modified ELISA techniques. This microT microM-ELISA method may be adapted for use with other antigens for which specific antibodies are available. These methods were used to compare cartilage and muscle differentiation in 1 to 4 d microT microM cultures grown in serum-containing (SCM) and defined (DM) media. The DM contains minimal additives (insulin, hydrocortisone, and in some cases, ascorbate or transferrin) and supports both chondrogenesis and myogenesis. The colorimetric analyses agree well with the morphologic appraisal of chondrogenesis and myogenesis. Similar numbers of cartilage nodules formed in all cultures, but in DM the nodules failed to enlarge; explaining the reduced matrix synthesis in DM as compared with SCM, and suggesting that nodule enlargement is a discrete, serum-dependent step. Studies of selected additives to DM show that transferrin enhances myogenesis, ascorbic acid enhances chondrogenesis, and retinoic acid inhibits chondrogenesis. Together, the microT microM system, in situ colorimetric assays of chondrogenesis and myogenesis, and DM will allow rapid prescreening of teratogens and screening of various bioactive compounds (e.g., hormones, growth factors, vitamins, adhesion factors) for effects on limb mesenchymal cell differentiation.

  16. 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.

  17. Glycerol Monolaurate Antibacterial Activity in Broth and Biofilm Cultures

    PubMed Central

    Schlievert, Patrick M.; Peterson, Marnie L.

    2012-01-01

    Background Glycerol monolaurate (GML) is an antimicrobial agent that has potent activity against gram-positive bacteria. This study examines GML antibacterial activity in comparison to lauric acid, in broth cultures compared to biofilm cultures, and against a wide range of gram-positive, gram-negative, and non-gram staining bacteria. Methodology/Principal Findings GML is ≥200 times more effective than lauric acid in bactericidal activity, defined as a ≥3 log reduction in colony-forming units (CFU)/ml, against Staphylococcus aureus and Streptococcus pyogenes in broth cultures. Both molecules inhibit superantigen production by these organisms at concentrations that are not bactericidal. GML prevents biofilm formation by Staphylococcus aureus and Haemophilus influenzae, as representative gram-positive and gram-negative organisms, tested in 96 well microtiter plates, and simultaneously is bactericidal for both organisms in mature biofilms. GML is bactericidal for a wide range of potential bacterial pathogens, except for Pseudomonas aeruginosa and Enterobacteriaceae. In the presence of acidic pH and the cation chelator ethylene diamine tetraacetic acid, GML has greatly enhanced bactericidal activity for Pseudomonas aeruginosa and Enterobacteriaceae. Solubilization of GML in a nonaqueous delivery vehicle (related to K-Y Warming®) enhances its bactericidal activity against S. aureus. Both R and S, and 1 and 2 position lauric acid derivatives of GML exhibit bactericidal activity. Despite year-long passage of Staphylococcus aureus on sub-growth inhibitory concentrations of GML (0.5 x minimum bactericidal concentration), resistance to GML did not develop. Conclusions/Significance GML may be useful as a broad-spectrum human or animal topical microbicide and may be useful as an environmental surface microbicide for management of bacterial infections and contamination. PMID:22808139

  18. Organoid Culture of Isolated Cells from Patient-derived Tissues with Colorectal Cancer

    PubMed Central

    Xie, Bing-Ying; Wu, Ai-Wen

    2016-01-01

    Background: Colorectal cancer (CRC) is a heterogeneous disease; current research relies on cancer cell lines and animal cancer models, which may not precisely imitate inner human tumors and guide clinical medicine. The purpose of our study was to explore and further improve the process of producing three-dimensional (3D) organoid model and impel the development of personalized therapy. Methods: We subcutaneously injected surgically resected CRC tissues from a patient into BALB/c-nu mice to build patient-derived xenografts (PDXs). Isolated cells from PDXs at appropriate tumor size were mingled with Matrigel, and then seeded in ultra-low attachment 96-well plates at four cell densities (500, 1000, 2000, and 4000 single cells/well). Cells were cultured with advanced Dulbecco's Modified Eagle Medium/F12 medium additional with various factors added to maintain tumor's biological traits and growth activity. The growth curves of the four cell densities were measured after 24 h of culture until 25 days. We evaluated the effects of four chemotherapeutic agents on organoid model by the CellTiter-Glo® Luminescent Cell Viability Assay. Hematoxylin and eosin (H and E) staining of 3D organoids was performed and compared with patient and CRC PDX tissues. Furthermore, immunohistochemistry was performed, in which the organoids were stained with the proliferation marker, Ki-67. During the experimental process, a phase-contrast microscope was used. Results: Phenotype experimental results showed that 3D organoids were tightly packed together and grew robustly over time. All four densities of cells formed organoids while that composed of 2000 cells/well provided an adequate cultivation system and grew approximately 8-fold at the 25th day. The chemosensitivity of the four conventional drugs was [s]-10-hydroxycamptothecin > mitomycin C > adriamycin > paclitaxel, which can guide clinical treatment. Histological features of CRC patient's tumor tissues and mice tumor xenograft tissues were

  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. 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.

  1. Obliquity along plate boundaries

    NASA Astrophysics Data System (ADS)

    Philippon, Mélody; Corti, Giacomo

    2016-12-01

    Most of the plate boundaries are activated obliquely with respect to the direction of far field stresses, as roughly only 8% of the plate boundaries total length shows a very low obliquity (ranging from 0 to 10°, sub-orthogonal to the plate displacement). The obliquity along plate boundaries is controlled by (i) lateral rheological variations within the lithosphere and (ii) consistency with the global plate circuit. Indeed, plate tectonics and magmatism drive rheological changes within the lithosphere and consequently influence strain localization. Geodynamical evolution controls large-scale mantle convection and plate formation, consumption, and re-organization, thus triggering plate kinematics variations, and the adjustment and re-orientation of far field stresses. These geological processes may thus result in plate boundaries that are not perpendicular but oblique to the direction of far field stresses. This paper reviews the global patterns of obliquity along plate boundaries. Using GPlate, we provide a statistical analysis of present-day obliquity along plate boundaries. Within this framework, by comparing natural examples and geological models, we discuss deformation patterns and kinematics recorded along oblique plate boundaries.

  2. Plating Tank Control Software

    SciTech Connect

    Krafcik, John

    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.

  3. 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.

  4. Characterization of antibody binding to cell surface antigens using a plasma membrane-bound plate assay.

    PubMed

    Vater, C A; Reid, K; Bartle, L M; Goldmacher, V S

    1995-01-01

    A procedure has been developed for measuring antibody binding to cell surface antigens using an immobilized plasma membrane fraction. In this method, isolated plasma membranes are dried onto wells of a 96-well microtiter plate and incubated with antibodies that recognize a cell surface protein. Bound antibody is detected indirectly using an enzyme-linked or fluorescently tagged second antibody. Alternatively, the primary antibody itself can be labeled and its binding can be detected directly. The assay is simple and fast and provides several advantages over whole cell binding assays currently in widespread use.

  5. Malachite green photosensitive plates.

    PubMed

    Solano, C

    1989-08-15

    An experimental study of the behavior of malachite green sensitized plates was carried out. The transmittance variation of the irradiated plates was taken as a parameter. It has been observed that photoreduction in the malachite green plates is present only when ammonium dichromate is added to the plates. The introduction of external electron donors does not improve the photochemical reaction. It has been determined that malachite green molecules form a weak complex with the dichromate molecules and this complex can only be destroyed photochemically. This effect can explain the limited response of the malachite green dichromated plates.

  6. 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.

  7. 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.

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

    PubMed

    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.

  9. Human CD34(+) progenitor hematopoiesis in liquid culture for in vitro assessment of drug-induced myelotoxicity.

    PubMed

    Guo, Liang; Hamre, John; Davis, Myrtle; Parchment, Ralph E

    2016-03-01

    Utilization of validated CFU-GM assays for myelotoxicity screening is hampered by its labor-intensive and low-throughput nature. Herein, we transformed the defined CFU-GM assay conditions and IC90 endpoint into a higher throughput format. Human CD34(+) hematopoietic progenitors were cultured in a 96-well plate for 14 days with the same cytokine (rhGM-CSF) used in the CFU-GM assay. Expansion and differentiation toward myeloid lineages were manifested by characteristic changes in nuclear and cytoplasmic morphology and by temporal expression patterns of CD34, CD11b and CD13 markers. Inhibition of CD34(+) cell myelopoiesis by 12 anticancer drugs known to induce myelotoxicity in the clinic was quantifiable using either general cytotoxicity endpoints (cell growth area or total nucleus count) or lineage specific readouts (count of cells expressing CD11b and/or CD13). The IC50 and IC90 values derived from the concentration-response curves of 14-day drug exposure in CD34(+) cell culture were highly correlated with those from the international validation study of the CFU-GM assay, demonstrating capability to assess general cytotoxicity, cell proliferation and myelopoiesis simultaneously. These results suggest that this human CD34(+) hematopoietic progenitor cell assay can be used as a direct replacement for the validated, low throughput CFU-GM assay, and could expand application of in vitro myelotoxicity testing.

  10. High-Throughput, Automated Protein A Purification Platform with Multiattribute LC-MS Analysis for Advanced Cell Culture Process Monitoring.

    PubMed

    Dong, Jia; Migliore, Nicole; Mehrman, Steven J; Cunningham, John; Lewis, Michael J; Hu, Ping

    2016-09-06

    The levels of many product related variants observed during the production of monoclonal antibodies are dependent on control of the manufacturing process, especially the cell culture process. However, it is difficult to characterize samples pulled from the bioreactor due to the low levels of product during the early stages of the process and the high levels of interfering reagents. Furthermore, analytical results are often not available for several days, which slows the process development cycle and prevents "real time" adjustments to the manufacturing process. To reduce the delay and enhance our ability to achieve quality targets, we have developed a low-volume, high-throughput, and high-content analytical platform for at-line product quality analysis. This workflow includes an automated, 96-well plate protein A purification step to isolate antibody product from the cell culture fermentation broth, followed by rapid, multiattribute LC-MS analysis. We have demonstrated quantitative correlations between particular process parameters with the levels of glycosylated and glycated species in a series of small scale experiments, but the platform could be used to monitor other attributes and applied across the biopharmaceutical industry.

  11. 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.

  12. 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.

  13. 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.

  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. 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.

  16. 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). 

  17. 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.

  18. 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.

  19. Radioisotopic quantitation in microtitration plates by an autofluorographic method.

    PubMed

    Freshney, R I; Morgan, D

    1978-07-01

    An autofluorographic record of drug titrations in microtitration plates has been obtained using cultures labelled with [35s] methionine after treatment with cytostatic drugs. By adding scintillation fluid directly to each culture well of the microtitration plate, and then centrifuging the evaporate the toluene and leave a flat even film of fluor, in situ scintillation is produced. When X-ray film is exposed to the plate the scintillation of individual wells is recorded as spots on the film. These may be interpreted by eye or by scanning densitometry.

  20. 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

  1. Pixelated neutron image plates

    NASA Astrophysics Data System (ADS)

    Schlapp, M.; Conrad, H.; von Seggern, H.

    2004-09-01

    Neutron image plates (NIPs) have found widespread application as neutron detectors for single-crystal and powder diffraction, small-angle scattering and tomography. After neutron exposure, the image plate can be read out by scanning with a laser. Commercially available NIPs consist of a powder mixture of BaFBr : Eu2+ and Gd2O3 dispersed in a polymer matrix and supported by a flexible polymer sheet. Since BaFBr : Eu2+ is an excellent x-ray storage phosphor, these NIPs are particularly sensitive to ggr-radiation, which is always present as a background radiation in neutron experiments. In this work we present results on NIPs consisting of KCl : Eu2+ and LiF that were fabricated into ceramic image plates in which the alkali halides act as a self-supporting matrix without the necessity for using a polymeric binder. An advantage of this type of NIP is the significantly reduced ggr-sensitivity. However, the much lower neutron absorption cross section of LiF compared with Gd2O3 demands a thicker image plate for obtaining comparable neutron absorption. The greater thickness of the NIP inevitably leads to a loss in spatial resolution of the image plate. However, this reduction in resolution can be restricted by a novel image plate concept in which a ceramic structure with square cells (referred to as a 'honeycomb') is embedded in the NIP, resulting in a pixelated image plate. In such a NIP the read-out light is confined to the particular illuminated pixel, decoupling the spatial resolution from the optical properties of the image plate material and morphology. In this work, a comparison of experimentally determined and simulated spatial resolutions of pixelated and unstructured image plates for a fixed read-out laser intensity is presented, as well as simulations of the properties of these NIPs at higher laser powers.

  2. 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.

  3. Designing Assemblies Of Plates

    NASA Technical Reports Server (NTRS)

    Williams, F. W.; Kennedy, D.; Butler, R.; Aston, G.; Anderson, M. S.

    1992-01-01

    VICONOPT calculates vibrations and instabilities of assemblies of prismatic plates. Designed for efficient, accurate analysis of buckling and vibration, and for optimum design of panels of composite materials. Written in FORTRAN 77.

  4. 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.

  5. 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.

  6. 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)

  7. Reduction of astrometric plates

    NASA Technical Reports Server (NTRS)

    Stock, J.

    1984-01-01

    A rapid and accurate method for the reduction of comet or asteroid plates is described. Projection equations, scale length correction, rotation of coordinates, linearization, the search for additional reference stars, and the final solution are examined.

  8. What's On Your Plate?

    MedlinePlus

    ... Table of Contents What's On Your Plate? Smart Food Choices for Healthy Aging www.nia.nih.gov/health/ ... calories in" and "calories out," and making good food choices as you age. Shopping Tips See how planning ...

  9. Feynman's wobbling plate

    NASA Astrophysics Data System (ADS)

    Tuleja, Slavomir; Gazovic, Boris; Tomori, Alexander; Hanc, Jozef

    2007-03-01

    In the book Surely You Are Joking, Mr. Feynman! Richard Feynman tells a story of a Cornell cafeteria plate being tossed into the air. As the plate spun, it wobbled. Feynman noticed a relation between the two motions. He solved the motion of the plate by using the Lagrangian approach. This solution didn't satisfy him. He wanted to understand the motion of the plate by analyzing the motion of its individual particles and the forces acting on them. He was successful, but he didn't tell us how he did it. We provide an elementary explanation for the two-to-one ratio of wobble to spin frequencies, based on an analysis of the motion of the particles and the forces acting on them. We also demonstrate the power of numerical simulation and computer animation to provide insight into a physical phenomenon and guidance on how to do the analysis.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Use of monocyte/endothelial cell co-cultures (in vitro) and a subcutaneous implant mouse model (in vivo) to evaluate a degradable polar hydrophobic ionic polyurethane.

    PubMed

    McDonald, Sarah M; Matheson, Loren A; McBane, Joanne E; Kuraitis, Drew; Suuronen, Erik; Santerre, Joseph Paul; Labow, Rosalind S

    2011-12-01

    Potential benefits of co-culturing monocytes (MC) with vascular smooth muscle cells have been reported on for tissue engineering applications with a degradable, polar, hydrophobic, and ionic polyurethane (D-PHI). Since the interaction of MC and endothelial cells (EC) within the blood vessel endothelium is also a process of wound repair it was of interest to investigate their function when cultured on the synthetic D-PHI materials, prior to considering the materials' use in vascular engineering. The co-culture (MC/EC) in vitro studies were carried out on films in 96 well plates and porous scaffold disks were prepared for implant studies in an in vivo subcutaneous mouse model. After 7 days in culture, the MC/EC condition was equal to EC growth but had lower esterase activity (a measure of degradative potential), no pro-inflammatory TNF-α and a relatively high anti-inflammatory IL-10 release while the ECs maintained their functional marker CD31. After explantation of the porous scaffolds, a live/dead stain showed that the cells infiltrating the scaffolds were viable and histological stains (May-Grunwald, Trichrome) demonstrated tissue in growth and extracellular matrix synthesis. Lysates from the implant scaffolds analyzed with a cytokine antibody array showed decreased pro-inflammatory cytokines (IL-6, TNF-α, GM-CSF), increased anti-inflammatory cytokines (IL-10, IL-13, TNF-RI), and increased chemotactic cytokines (MCP-1, MCP-5, RANTES). The low foreign body response elicited by D-PHI when implanted in vivo supported the in vitro studies (EC and MC co-culture), demonstrating that D-PHI promoted EC growth along with an anti-inflammatory MC, further demonstrating its potential as a tissue engineering scaffold for vascular applications.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. License plate detection algorithm

    NASA Astrophysics Data System (ADS)

    Broitman, Michael; Klopovsky, Yuri; Silinskis, Normunds

    2013-12-01

    A novel algorithm for vehicle license plates localization is proposed. The algorithm is based on pixel intensity transition gradient analysis. Near to 2500 natural-scene gray-level vehicle images of different backgrounds and ambient illumination was tested. The best set of algorithm's parameters produces detection rate up to 0.94. Taking into account abnormal camera location during our tests and therefore geometrical distortion and troubles from trees this result could be considered as passable. Correlation between source data, such as license Plate dimensions and texture, cameras location and others, and parameters of algorithm were also defined.

  2. Intermittent Plate Tectonics

    NASA Astrophysics Data System (ADS)

    Silver, P. G.; Behn, M. D.

    2006-12-01

    Intermittent Plate Tectonics A basic premise of Earth Science is that plate tectonics has been continuously operating since it began early in Earth's history. Yet, plate-tectonic theory itself, specifically the collisional phase of the Wilson Cycle, constitutes a process that is capable of stopping all plate motion. The plausibility of a plate-tectonic hiatus is most easily illustrated by considering the expected future of the present-day plate-tectonic configuration. Since the opening of the Atlantic at ~200 ma, the area of the Atlantic basin has been growing at the expense of the Pacific. If this trend continues, relative plate motion models predict that in ~350 my, the Pacific Ocean basin will effectively close leading to widespread continent-continent collisions. Since a continent-continent collision represents the termination of subduction locally, the accumulated effect of all collisions is to stop subduction globally. In this scenario, ridges would then stop spreading and young oceanic lithosphere would cool, reaching a steady-state thickness of 100 km in about 80 my, based on the properties of oceanic lithosphere today. This would constitute the stoppage of plate tectonics. The presumption that plate tectonics never stops in the face of continental collisions is equivalent to requiring that subduction flux is approximately constant through time, such that subduction initiation roughly balances subduction termination. Such a balance then raises several questions about the subduction initiation process. When and how does subduction initiate? Is there a detectible relationship between subduction cessation and subduction initiation? We can gain some guidance into these questions by examining the plate motion history over the last 200 my. Subduction initiation has occurred over the last 80 my in three intra- oceanic subduction zones: Aleutians, Marianas-Izu-Bonin and Tonga-Kermadec in the Pacific basin. In these cases, however, subduction initiation would not

  3. Nuclear reactor alignment plate configuration

    SciTech Connect

    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.

  4. 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…

  5. 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).

  6. 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 ...

  7. 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.

  8. A study of frog muscle maintained in organ culture

    PubMed Central

    Harris, A. J.; Miledi, R.

    1972-01-01

    1. Frog muscles are isolated and maintained in organ culture conditions for periods of up to 2 months. During the first 2 weeks, muscle fibres have normal resting membrane and action potentials. Subsequently the potentials decline in amplitude. 2. Slow muscle fibres also survive in culture and retain their ability to give maintained contractures. 3. Muscle sensory receptors continue to function in culture until the axon terminals degenerate at about 2 weeks. 4. Neuromuscular transmission is normal during the first few days of culture, after which the motor endings degenerate. Transmission persists longer (up to 17 days) if a long segment of nerve is left attached to the muscle. With short-nerve preparations failure of transmission in vivo occurs at about the same time as in culture. With long-nerve preparations failure of transmission is delayed even further in culture. 5. In short-nerve preparations miniature end-plate potentials disappear, in general, at about the time that transmission fails. In long-nerve preparations some end-plates continue to have miniature end-plate potential activity for a short time after nerve impulses cease to evoke any response; but eventually miniature potential activity disappears from all end-plates. 6. After a few days of electrical silence, miniature end-plate potentials reappear at some of the denervated end-plates. The proportion of denervated end-plates which show miniature end-plate potentials in culture is smaller than in muscles denervated in situ. 7. Electron microscopy shows that muscle structure is well preserved in culture, that the axons degenerate and that the Schwann cells move to occupy the space vacated by the axons. The Schwann cells are very probably the source of the acetylcholine which evokes miniature potentials in the denervated end-plates. ImagesPlate 3Plate 4Plate 5Plate 6Plate 7Plate 1Plate 2 PMID:4335913

  9. What Are Growth Plate Injuries?

    MedlinePlus

    ... nih.gov November 2014 What Are Growth Plate Injuries? Fast Facts: An Easy-to-Read Series of Publications ... Some inherited disorders 1 What Are Growth Plate Injuries? Fast Facts: An Easy-to-Read Series of Publications ...

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. Mechanisms in Thermal Mechanical Forming of Plates.

    DTIC Science & Technology

    1987-05-01

    specific locations within the plate is discussed, and recommendations for further research are made. Keywords: Metal plates; Ship plates; Material forming; Thermomechanics; Edge effect ; Laser line heating.

  16. 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.

  17. 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.

  18. 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

  19. Electronic Equipment Cold Plates

    DTIC Science & Technology

    1976-04-01

    equations for such a flow regiae. For laainar flow and Moderate teaperature differwwe« between the well «nd coolant, a aodifled Sieder -Tate...con- figuration. The heat-transfer coefficients, therefore, were determined by using both the Sieder -Tate and McAdams equations and the coaputed...values used In the analytical predictions. As with th* previous cold Plates, the Sieder -Tate equation gave too low of values for the heat- transfer

  20. The Plate Overlap Technique.

    DTIC Science & Technology

    1978-07-31

    INTRODUCTION 1 II. NOTATION 2 III. THE GNOMONIC PROJECTION 4 IV . THE PLATE OVERLAP TECHNIQUE 6 A. MOTIVATION 6 B. FORNULATION 9 C. ON STATISTICAL RIGOR 14 D...and new hardware. Since this aim was clearly recognized long ago, wherever possible in earlier documents or software development flexibility was...reader should see 1, 2, and 3. The procedures one should use to update stellar positions are discussed in 4 with applica- tions to the SAOC in 5. Non

  1. 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.

  2. Armor Plate Surface Roughness Measurements

    DTIC Science & Technology

    2005-04-01

    Armor Plate Surface Roughness Measurements by Brian Stanton, William Coburn, and Thomas J. Pizzillo ARL-TR-3498 April 2005... Armor Plate Surface Roughness Measurements Brian Stanton, William Coburn and Thomas J. Pizzillo Sensors and Electron Devices Directorate...October 2004 5a. CONTRACT NUMBER 5b. GRANT NUMBER 4. TITLE AND SUBTITLE Armor Plate Surface Roughness Measurements 5c. PROGRAM ELEMENT NUMBER

  3. 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.

  4. 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.

  5. Hypervelocity impact on shielded plates

    NASA Technical Reports Server (NTRS)

    Smith, James P.

    1993-01-01

    A ballistic limit equation for hypervelocity impact on thin plates is derived analytically. This equation applies to cases of impulsive impact on a plate that is protected by a multi-shock shield, and it is valid in the range of velocity above 6 km/s. Experimental tests were conducted at the NASA Johnson Space Center on square aluminum plates. Comparing the center deflections of these plates with the theoretical deflections of a rigid-plastic plate subjected to a blast load, one determines the dynamic yield strength of the plate material. The analysis is based on a theory for the expansion of the fragmented projectile and on a simple failure criterion. Curves are presented for the critical projectile radius versus the projectile velocity, and for the critical plate thickness versus the velocity. These curves are in good agreement with curves that have been generated empirically.

  6. 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.

  7. Perfused multiwell plate for 3D liver tissue engineering.

    PubMed

    Domansky, Karel; Inman, Walker; Serdy, James; Dash, Ajit; Lim, Matthew H M; Griffith, Linda G

    2010-01-07

    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 of 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.

  8. 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.

  9. 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.

  10. Lipophilic rather than hydrophilic photosensitizers show strong adherence to standard cell culture microplates under cell-free conditions.

    PubMed

    Engelhardt, Victoria; Kiesslich, Tobias; Berlanda, Juergen; Hofbauer, Stefanie; Krammer, Barbara; Plaetzer, Kristjan

    2011-06-02

    Analysis of photosensitizer (PS) uptake kinetics into tumor cells is a standard cell culture experiment in photodynamic therapy (PDT) - usually performed in plastic microplates or petri dishes. Organic substances such as PS can potentially interact with the plastic surfaces. In this study, we provide a qualitative comparison of three lipophilic PS (hypericin, Foscan® and Photofrin®) and two rather hydrophilic PS formulations (PVP-hypericin and aluminum (III) phthalocyanine tetrasulfonate chloride) regarding their adherence to the surfaces of 96-well microplates obtained from four different manufacturers. For estimation of the relevance of PS adherence for cellular uptake studies we compared the fluorescence signal of the respective PS in microplates containing A431 human epithelial carcinoma cells with microplates incubated with the respective PS under cell-free conditions. We demonstrate that lipophilic PS substances show a strong adherence to microplates - in case of direct lysis and fluorescence measurement resulting in 50% up to 90% of the overall signal to be caused by adherence of the substances to the plastic materials in a cellular uptake experiment. For the hydrophilic compounds, adherence is negligible. Interestingly, adherence of PS agents to microplates takes place in a time-dependent and thus kinetic-like manner, requiring up to several hours to reach a plateau of the fluorescence signal. Furthermore, PS adherence is a function of the PS concentration applied and no saturation effect was observed for the concentrations used in this study. Taken together, this study provides a systematic analysis under which conditions PS adherence to cell culture plates may contribute to the overall fluorescence signal in - for example - PS uptake experiments.

  11. Earth's Decelerating Tectonic Plates (Invited)

    NASA Astrophysics Data System (ADS)

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

    2009-12-01

    We employ a recently developed convection model constrained by seismic, geodynamic and mineral physics data (Simmons et al., GJI 2009) to explore the impact of time-dependent changes in mantle buoyancy forces on tectonic plate accelerations. This plate-coupled mantle convection model incorporates a viscosity structure that reconciles both glacial isostatic adjustment and global convection-related data sets (Mitrovica & Forte, EPSL 2004) and it successfully reproduces present-day plate velocities, global surface gravity and topography data. This convection model predicts the recent deceleration of several major plates in the Pacific and Indo-Atlantic hemispheres. Independent verification of these predictions is a fundamental test of the plausibility of the buoyancy forces and rheological structure in the convection model. To this end, we consider marine magnetic anomaly and space geodetic constraints on tectonic plate motions to determine a new global map of present-day rates of change of plate velocities. This map shows that several major plates, such as the Pacific, Africa and Nazca plates are presently decelerating and that they contribute to a globally-averaged slowdown in tectonic plate speeds. These joint geologic-geodetic inferences of plate decelerations are consistent with those predicted by our tomography-based convection model.

  12. 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

    2016-01-01

    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.

  13. Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

    PubMed

    Bego, Mariana G; Côté, Édouard A; Cohen, Éric A

    2015-09-01

    HIV-1 innate sensing requires direct contact of infected CD4+ T cells with plasmacytoid dendritic cells (pDCs). In order to study this process, the protocols described here use freshly isolated human peripheral blood mononuclear cells (PBMCs) or plasmacytoid dendritic cells (pDCs) to sense infections in either T cell line (MT4) or heterologous primary CD4+ T cells. In order to ensure proper sensing, it is essential that PBMC are isolated immediately after blood collection and that optimal percentage of infected T cells are used. Furthermore, multi-parametric flow cytometric staining can be used to confirm that PBMC samples contain the different cell lineages at physiological ratios. A number of controls can also be included to evaluate viability and functionality of pDCs. These include, the presence of specific surface markers, assessing cellular responses to known agonist of Toll-Like Receptors (TLR) pathways, and confirming a lack of spontaneous type-I interferon (IFN) production. In this system, freshly isolated PBMCs or pDCs are co-cultured with HIV-1 infected cells in 96 well plates for 18-22 hr. Supernatants from these co-cultures are then used to determine the levels of bioactive type-I IFNs by monitoring the activation of the ISGF3 pathway in HEK-Blue IFN-α/β cells. Prior and during co-culture conditions, target cells can be subjected to flow cytometric analysis to determine a number of parameters, including the percentage of infected cells, levels of specific surface markers, and differential killing of infected cells. Although, these protocols were initially developed to follow type-I IFN production, they could potentially be used to study other imuno-modulatory molecules released from pDCs and to gain further insight into the molecular mechanisms governing HIV-1 innate sensing.

  14. Plating on difficult-to-plate metals: what's new

    SciTech Connect

    Wiesner, H.J.

    1980-07-30

    Some of the changes since 1970 in procedures for plating on such materials as titanium, molybdenum, silicon, aluminum, and gallium arsenide are summarized. While basic procedures for plating some of these materials were developed as many as 30 to 40 years ago, changes in the end uses of the plated products have necessitated new plating processes. In some cases, vacuum techniques - such as ion bombardment, ion implantation, and vacuum metallization - have been introduced to improve the adhesion of electrodeposits. In other cases, these techniques have been used to deposit materials upon which electrodeposits are required.

  15. 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.

  16. 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.

  17. Antiproliferative effect of berberine on canine mammary gland cancer cell culture.

    PubMed

    Sefidabi, Reyhaneh; Mortazavi, Pejman; Hosseini, Saeed

    2017-01-01

    Canine mammary gland tumors are the most frequent cause of cancer in female dogs. Numerous studies using cancer cell lines and clinical trials have indicated that various natural products and antioxidants reduce or possibly prevent the development of cancer. Berberine (BBR), the most important alkaloid in the Berberidaceae, which exerts a wide range of pharmacological and biochemical effects, has drawn much attention due to its particularly high antitumor activity in vitro and in animal studies. The aim of the present study was to investigate the antiproliferative effect of BBR against a canine mammary gland carcinoma cell line (CF41.Mg) in vitro. CF41.Mg cells were cultured in RPMI-1640 medium containing 10% heat inactived fetal bovine serum (FBS) and 100 mg/ml peniciline-streptomycin. Subsequently the cells were treated with different concentrations of BBR chloride (10, 25, 50, 100 and 200 µM) at a density of 12,000 cells/well in 96-well plates. Following treatment, the MTT assay was used to detect cell viability after 24-, 48- and 72-h incubations at 37°C with 5% CO2. The results indicated that BBR inhibited proliferation of canine mammary gland carcinoma cells, as treatment with 100 µM BBR for 24 h resulted in a significant decrease in cell viability (P<0.005). As the present study demonstrated that BBR (10-200 µM) induced cancer cell death, it is proposed that BBR may serve as a candidate agent against canine mammary tumor cells via its antiproliferative activity.

  18. In vitro stimulation of bovine peripheral blood lymphocytes: comparison of round- and flat-bottom microtiter plates for detection of tuberculin hypersensitivity.

    PubMed

    Sloane, E D; Muscoplat, C C; Kaneene, J M; Klausner, D J; Thoen, C O; Johnson, D W

    1978-02-01

    Lymphocytes from Mycobacterium bovis-sensitized and normal cattle were cultured in round- and/or flat-bottom microtiter plates and stimulated with M. bovis purified protein derivative (PPD) tuberculin. Blastogenic responses of lymphocytes from M. bovis-sensitized cattle to PPD cultured in round-bottom plates were significantly greater than those of lymphocytes cultured in flat-bottom microtiter plates. Normal lymphocytes of nonsensitized cattle were not stimulated by PPD in either round- or flat-bottom microtiter plates. Kinetics of lymphocyte responses in round-bottom plates are presented.

  19. 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.

  20. Dissolution of HFIR control plates

    SciTech Connect

    Posey, J.C.

    1984-03-01

    A process was developed for the dissolution of High Flux Isotope Reactor (HFIR) control plates. These plates consist of aluminum metal, intensely radioactive europium oxide, and a small amount of tantalum metal. The radioactive solution will be diluted, mixed with grout, and disposed of by shale fracture. The plates are dissolved in nitric acid using a mercury catalyst. Conditions were determined that would produce a reaction rate compatible with existing equipment. 3 references, 1 figure, 3 tables.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. Polymicrobial Chronic Infection Including Acinetobacter Baumannii in a Plated Segmental Defect in the Rat Femur

    DTIC Science & Technology

    2008-01-01

    Including Acinetobacter baumannii in a Plated Segmental Defect in the Rat Femur PRINCIPAL INVESTIGATOR: Dean T. Tsukayama, MD...FEB 2007 - 31 DEC 2007 4. TITLE AND SUBTITLE Polymicrobial Chronic Infection Including Acinetobacter baumannii 5a. CONTRACT NUMBER in a Plated...bone isolate of Acinetobacter baumannii exhibited very little osteolytic involvement when used alone in the model. Qualitative cultures indicated very

  8. 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.

  9. Intermittent plate tectonics?

    PubMed

    Silver, Paul G; Behn, Mark D

    2008-01-04

    Although it is commonly assumed that subduction has operated continuously on Earth without interruption, subduction zones are routinely terminated by ocean closure and supercontinent assembly. Under certain circumstances, this could lead to a dramatic loss of subduction, globally. Closure of a Pacific-type basin, for example, would eliminate most subduction, unless this loss were compensated for by comparable subduction initiation elsewhere. Given the evidence for Pacific-type closure in Earth's past, the absence of a direct mechanism for termination/initiation compensation, and recent data supporting a minimum in subduction flux in the Mesoproterozoic, we hypothesize that dramatic reductions or temporary cessations of subduction have occurred in Earth's history. Such deviations in the continuity of plate tectonics have important consequences for Earth's thermal and continental evolution.

  10. 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

  11. Downgoing plate controls on overriding plate deformation in subduction zones

    NASA Astrophysics Data System (ADS)

    Garel, Fanny; Davies, Rhodri; Goes, Saskia; Davies, Huw; Kramer, Stephan; Wilson, Cian

    2014-05-01

    Although subduction zones are convergent margins, deformation in the upper plate can be extensional or compressional and tends to change through time, sometimes in repeated episodes of strong deformation, e.g, phases of back-arc extension. It is not well understood what factors control this upper plate deformation. We use the code Fluidity, which uses an adaptive mesh and a free-surface formulation, to model a two-plate subduction system in 2-D. The model includes a composite temperature- and stress-dependent rheology, and plates are decoupled by a weak layer, which allows for free trench motion. We investigate the evolution of the state of stress and topography of the overriding plate during the different phases of the subduction process: onset of subduction, free-fall sinking in the upper mantle and interaction of the slab with the transition zone, here represented by a viscosity contrast between upper and lower mantle. We focus on (i) how overriding plate deformation varies with subducting plate age; (ii) how spontaneous and episodic back-arc spreading develops for some subduction settings; (iii) the correlation between overriding plate deformation and slab interaction with the transition zone; (iv) whether these trends resemble observations on Earth.

  12. A human CXCL13-induced actin polymerization assay measured by fluorescence plate reader.

    PubMed

    Alley, Jennifer; Bloom, Laird; Kasaian, Marion; Gao, Huilan; Berstein, Gabriel; Clark, James D; Miao, Wenyan

    2010-02-01

    The chemokine receptor CXCR5 is predominantly expressed on mature B cells and follicular T-helper cells. CXCR5 and its ligand CXCL13 participate in ectopic germinal center formation at the inflammatory sites of multiple immune diseases such as rheumatoid arthritis, multiple sclerosis, and Sjogren's syndrome. Therefore, disrupting CXCL13-induced chemotaxis may be a fruitful approach for developing therapeutics in treating these diseases. Cells undergo cytoskeletal rearrangement prior to chemotaxis, and therefore actin polymerization can be used as a surrogate readout more proximal to chemokine receptor activation than chemotaxis. Conventionally, actin polymerization is measured by fluorescence microscopy or flow cytometry, which are either of low throughput or in need of special instruments. We developed a 96-well actin polymerization assay that can process 1,000 to 1,500 samples a day. This assay uses a standard laboratory fluorescence microplate reader as the detection instrument and was optimized for various experimental conditions such as cell density, actin filament staining reagent, staining buffer, and cell culture conditions. We demonstrate that this actin polymerization assay in 96-well format exhibits the expected pharmacology for human CXCR5 and is suitable as a primary functional assay to screen neutralizing scFv in crude bacterial peri-preps and a secondary assay for small compound collections.

  13. 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.

  14. Metals plated on fluorocarbon polymers

    NASA Technical Reports Server (NTRS)

    Ford, H.; Krasinsky, J. B.; Vango, S. P.

    1964-01-01

    Electroplating lead on fluorocarbon polymer parts is accomplished by etching the parts to be plated with sodium, followed by successive depositions of silver and lead from ultrasonically agitated plating solutions. Metals other than lead may be electroplated on the silvered parts.

  15. Metal vapor arc ion plating

    DOEpatents

    Bertram, L.A.; Fisher, R.W.; Mattox, D.M.; Zanner, F.J.

    1986-09-09

    A method and apparatus for ion plating are described. The apparatus uses more negative than a first electrode voltage in a vacuum arc remelt system to attract low energy ions from the anode electrode to the article to be plated. 2 figs.

  16. Gold plating on spectacle frames.

    PubMed

    Kenny, I; Mitchell, J W; Walsh, G

    1997-05-01

    An investigation was carried out into the thickness and standard of application of the plating and lacquer coatings applied to three metal spectacle frames. All conform to BS 6625 (1991) for plating thickness, but there was considerable variation in regularity and porosity.

  17. 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.

  18. 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

    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 (r (2) = 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.

  19. 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.

  20. The moving plate capacitor paradox

    NASA Astrophysics Data System (ADS)

    Davis, B. R.; Abbott, D.; Parrondo, J. M. R.

    2000-03-01

    For the first time we describe an apparent paradox concerning a moving plate capacitor driven by thermal noise from a resistor. A demon restores the plates of the capacitor to their original position, only when the voltage across the capacitor is small—hence only small forces are present for the demon to work against. The demon has to work harder than this to avoid the situation of perpetual motion, but the question is how? We explore the concept of a moving plate capacitor, driven by noise, a step further by examining the case where the restoring force on the capacitor plates is provided by a simple spring, rather than some unknown demon. We display simulation results with interesting behavior, particularly where the capacitor plates collide with each other.

  1. 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.

  2. Underwater electrical discharge in plate to plate configuration

    NASA Astrophysics Data System (ADS)

    Stelmashuk, Vitaliy

    2016-09-01

    Two main configurations of high voltage electrodes submersed in water have been used for an electrical discharge generation: pin to pin and pin to plate. An electrical breakdown between plate electrodes is generally difficult to reproduce, because there is a uniform and weak electric field. One major advantage of using plate electrodes is their greater ``wear hardness'' to high-energy discharges. The plate electrodes can withstand extremely high energy deposition at which the pin electrode is quickly destroyed. The electrical discharge between plate electrodes can be initiated by creating an inhomogeneity in the electrical field. Two methods of discharge initiation between plate electrodes are proposed for this aim: 1) focusing of a shock wave in the interelectrode region; 2) a bubble injection into the electrode gap. The shock wave creates favourable conditions for the electrical breakdown between the two plate electrodes: it causes that numerous microbubbles of dissolved air start to grow and serve as locations for streamer initiation. In the second method the gas bubble is injected from the one of the electrodes, which has a gas inlet hole on the lateral face for this purpose. A ``volcano'' like morphology of positive streamers are observed in the experiments with weak electric field. The authors are grateful to MEYS grant INGO LG 15013.

  3. 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.

  4. 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.

  5. 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.

  6. Smad4 regulates growth plate matrix production and chondrocyte polarity

    PubMed Central

    Whitaker, Amanda T.; Berthet, Ellora; Cantu, Andrea; Laird, Diana J.

    2017-01-01

    ABSTRACT Smad4 is an intracellular effector of the TGFβ family that has been implicated in Myhre syndrome, a skeletal dysplasia characterized by short stature, brachydactyly and stiff joints. The TGFβ pathway also plays a critical role in the development, organization and proliferation of the growth plate, although the exact mechanisms remain unclear. Skeletal phenotypes in Myhre syndrome overlap with processes regulated by the TGFβ pathway, including organization and proliferation of the growth plate and polarity of the chondrocyte. We used in vitro and in vivo models of Smad4 deficiency in chondrocytes to test the hypothesis that deregulated TGFβ signaling leads to aberrant extracellular matrix production and loss of chondrocyte polarity. Specifically, we evaluated growth plate chondrocyte polarity in tibiae of Col2-Cre+/−;Smad4fl/fl mice and in chondrocyte pellet cultures. In vitro and in vivo, Smad4 deficiency decreased aggrecan expression and increased MMP13 expression. Smad4 deficiency disrupted the balance of cartilage matrix synthesis and degradation, even though the sequential expression of growth plate chondrocyte markers was intact. Chondrocytes in Smad4-deficient growth plates also showed evidence of polarity defects, with impaired proliferation and ability to undergo the characteristic changes in shape, size and orientation as they differentiated from resting to hypertrophic chondrocytes. Therefore, we show that Smad4 controls chondrocyte proliferation, orientation, and hypertrophy and is important in regulating the extracellular matrix composition of the growth plate. PMID:28167493

  7. Quercetin treatment changes fluxes in the primary metabolism and increases culture longevity and recombinant α₁-antitrypsin production in human AGE1.HN cells.

    PubMed

    Niklas, Jens; Nonnenmacher, Yannic; Rose, Thomas; Sandig, Volker; Heinzle, Elmar

    2012-04-01

    Addition of the flavonoid quercetin to cultivations of the α(1)-antitrypsin (A1AT) producing human AGE1.HN.AAT cell line resulted in alterations of the cellular physiology and a remarkable improvement of the overall performance of these cells. In a first screening in 96-well plate format, toxicity and the effect of quercetin on the lactate/glucose ratio was analyzed. It was found that quercetin treatment reduced the lactate/glucose ratio dose dependently. An increase in culture longevity, viable cell density (160% of control), and A1AT concentration (from 0.39 g/L in the control to 0.76 g/L with quercetin, i.e., 195% of the control) was observed in batch cultivation with 10 μM quercetin compared to the control. A detailed analysis of quercetin effects on primary metabolism revealed dose-dependent alterations in metabolic fluxes. Quercetin addition resulted in an improved channeling of pyruvate into the mitochondria accompanied by reduced waste product formation and stimulation of TCA cycle activity. The observed changes in cellular physiology can be explained by different properties of quercetin and its metabolites, e.g., inhibition of specific enzymes, stimulation of oxidation of cytoplasmic, and mitochondrial NADH resulting in reduced NADH/NAD(+) ratio, and cytoprotective activity. The present study shows that the addition of specific effectors to the culture medium represents a promising strategy to improve the cellular metabolic phenotype and the production of biopharmaceuticals. The provided results contribute, additionally, to an improved understanding of quercetin action on the metabolism of human cells in a general physiological context.

  8. Rat liver clone-9 cells in culture as a model for screening hepatotoxic potential of food-related products: hepatotoxicity of deoxynivalenol.

    PubMed

    Sahu, Saura C; Garthoff, Larry H; Robl, Martin G; Chirtel, Stuart J; Ruggles, Dennis I; Flynn, Thomas J; Sobotka, Thomas J

    2008-08-01

    Deoxynivalenol (DON) is a mycotoxin food contaminant found in several cereal grains. The literature on the liver toxicity of DON in vivo is conflicting and does not clearly characterize its hepatotoxic effects. Cultured rat liver clone-9 cells were used as a model to assess the hepatotoxic potential of DON. The cell cultures, seeded onto 96-well plates, were treated at confluence with varying concentrations of DON (0-100 microg ml(-1)) for 48 h at 37 degrees C in 5% CO2. After the treatment period, the cells were assayed for a number of hepatotoxic endpoints that included cytotoxicity, double-stranded DNA (ds-DNA) content, oxidative stress and mitochondrial function. The concentration-dependent toxicity of DON, as measured by cytotoxicity and ds-DNA content, was observed over the entire concentration range studied beginning at 0.5 microg ml(-1). DON also induced a significant concentration-dependent increase in oxidative stress at DON concentrations starting at 10 microg ml(-1). The mitochondrial function of the treated cells decreased with the increasing concentration of DON exposure, but it was not statistically different from that of the control value. Liver histopathology observed at 3, 24 and 72 h following a single intraperitoneal administration dose of DON (10 mg kg(-1) BW) to adult male rats is consistent with early mild hepatotoxicity. The overall results of this study suggest that acute DON exposure has early mild cytotoxic effects on hepatocytes in vivo that are expressed as severe effects in rat liver clone-9 cells in vitro.

  9. 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.

  10. In vitro clonogenic capacity of periodontal ligament fibroblasts cultured with Emdogain.

    PubMed

    Ashkenazi, Malka; Shaked, Ilanit

    2006-02-01

    The aim of the present study was to evaluate the efficiency of Emdogain (EMD) in preserving the size of the periodontal ligament progenitor pool (clonogenic capacity) and in promoting their proliferation. Periodontal ligament fibroblasts (PDLF) were obtained from explants of young permanent healthy tooth. After initial outgrowth (10 days to 2 weeks following explantation), the culture medium of experimental flasks was replaced with medium supplemented with 100 microg ml(-1) EMD, whereas the other served as controls and were fed with regular medium. Following 5 weeks, the cells were washed (3x), harvested (trypsin + EDTA), and evaluated for their viability. Viable cells from each group were inoculated into six 96-well plates at a concentration of one viable cell per two wells and were allowed to grow for 5 weeks. The percentage of cells with clonogenic capacity was determined as the number of colonies formed/number of cells seeded x 100 in the experimental and control groups. Three degrees of dish area coverage were utilized: up to 25%, between 25% and 75% and higher than 75%. This experiment was repeated four times from four different donors. A total of 2328 cells were evaluated, half of which, were cultured with EMD. The mean percentage of cells (from all donors) who exhibited any clonogenic capacity in the presence of EMD was comparable with that of cells cultured in the absence of EMD: 26.6 +/- 14.3% when compared with 34.6 +/- 20.6% respectively (P = 0.186). Similarly, the percentage of clones that proliferated to cover up to 25% of the well area was comparable in the two groups 7.5 +/- 8.6 for EMD-treated clones and 7.1 +/- 7.8 for untreated clones (P = 0.674). The percentage of clones that proliferated to cover 25% up to 75% of the well area was greater EMD-treated clones as compared with the untreated cells: 8.1 +/- 6.7% vs 3.8 +/- 3%. However this difference was not statistically significant (P = 0.277). In contrast, the percentage of clones that covered

  11. Shockwaves Cause Synaptic Degeneration in Cultured Neurons

    DTIC Science & Technology

    2009-11-02

    constructed of delrin. A piezoresistive pressure sensor (Endevco Model 8530C) was mounted flush with the plate, coaxial with the center of the gene gun ...biolostic gene gun to deliver shockwaves to cultured hippocampal or cortical neurons. These cultured cells form abundant synapses in vitro, and after a 24-48...neurons, we used a biolostic gene gun to deliver shockwaves to cultured hippocampal or cortical neurons. These cultured cells form abundant synapses in

  12. Tectonics: Changing of the plates

    NASA Astrophysics Data System (ADS)

    Brandon, Alan

    2016-10-01

    The composition of Earth's crust depends on the style of plate tectonics and of the melting regimes in the mantle. Analyses of the oldest identified rocks suggest that these styles and the resulting crust have changed over Earth's history.

  13. 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.

  14. MyPlate Daily Checklist

    MedlinePlus

    ... Kids Making Family Mealtimes Fun Professionals Nutrition Communicators Network Communicator’s Guide Teachers Health Professionals MyPlate Graphics Multiple Languages En Español Other Languages Healthy Eating Style ...

  15. ChooseMyPlate.gov

    MedlinePlus

    ... Kids Making Family Mealtimes Fun Professionals Nutrition Communicators Network Communicator’s Guide Teachers Health Professionals MyPlate Graphics Multiple Languages En Español Other Languages Healthy Eating Style ...

  16. 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.

  17. Estimation of bisphenol A-Human toxicity by 3D cell culture arrays, high throughput alternatives to animal tests.

    PubMed

    Lee, Dong Woo; Oh, Woo-Yeon; Yi, Sang Hyun; Ku, Bosung; Lee, Moo-Yeal; Cho, Yoon Hee; Yang, Mihi

    2016-09-30

    Bisphenol A (BPA) has been widely used for manufacturing polycarbonate plastics and epoxy resins and has been extensively tested in animals to predict human toxicity. In order to reduce the use of animals for toxicity assessment and provide further accurate information on BPA toxicity in humans, we encapsulated Hep3B human hepatoma cells in alginate and cultured them in three dimensions (3D) on a micropillar chip coupled to a panel of metabolic enzymes on a microwell chip. As a result, we were able to assess the toxicity of BPA under various metabolic enzyme conditions using a high-throughput and micro assay; sample volumes were nearly 2,000 times less than that required for a 96-well plate. We applied a total of 28 different enzymes to each chip, including 10 cytochrome P450s (CYP450s), 10 UDP-glycosyltransferases (UGTs), 3 sulfotransferases (SULTs), alcohol dehydrogenase (ADH), and aldehyde dehydrogenase 2 (ALDH2). Phase I enzyme mixtures, phase II enzyme mixtures, and a combination of phase I and phase II enzymes were also applied to the chip. BPA toxicity was higher in samples containing CYP2E1 than controls, which contained no enzymes (IC50, 184±16μM and 270±25.8μM, respectively, p<0.01). However, BPA-induced toxicity was alleviated in the presence of ADH (IC50, 337±17.9μM), ALDH2 (335±13.9μM), and SULT1E1 (318±17.7μM) (p<0.05). CYP2E1-mediated cytotoxicity was confirmed by quantifying unmetabolized BPA using HPLC/FD. Therefore, we suggest the present micropillar/microwell chip platform as an effective alternative to animal testing for estimating BPA toxicity via human metabolic systems.

  18. Reflection-Zone-Plate Antenna

    NASA Technical Reports Server (NTRS)

    Franke, John M.; Leighty, Bradley D.

    1989-01-01

    Microwave antenna, based on reflection holography, designed and tested. Modified to produce arbitrary beam patterns by controlling relief pattern. Antenna planar or contoured to supporting structure. Low off-axis radar cross section at frequencies removed from operational frequency. Interference pattern produced by spherical wave intersecting plane wave consists of concentric circles similar to Newton's rings. Pattern identical to Fresnel zone plate, which has lens properties. Plane wave incident on hologram, or zone plate, focused to point.

  19. 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.

  20. How mantle slabs drive plate tectonics.

    PubMed

    Conrad, Clinton P; Lithgow-Bertelloni, Carolina

    2002-10-04

    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.

  1. Geometry of the Cocos Plate Under North American Plate

    NASA Astrophysics Data System (ADS)

    Perez-Campos, X.

    2015-12-01

    The Cocos plate subducts under the North American plate with a complex geometry, and previous seismicity studies revealed some of this complexity. However, details of the geometry and the depth that the plate penetrates werelargely unknown. Since 2004, temporary experiments and the expansion of the permanent network of the Servicio Sismológico Nacional (SSN, Mexican National Seismological Service) have improved resolution of the plate geometry and have helped to map its descent into the upper mantle. Going from northwest to southeast, the Cocos plate appears to be fragmenting into north and south segments. The north segment subducts with an angle of ~30º and the south with an angle of ~10-15º. The transition is smooth near the trench and progresses to a tear at depth; this coincides with the projection of the Orozco Fracture Zone to depth. Also, this transition marks the limit of the presence to the south of an ultra slow velocity layer (USL) on top of the slab.South of this transition, the Cocos plate subducts horizontally , underplating the North American plate for a distance of ~140 to ~300 km from the trench. Along this horizontal region, silent slow events (SSE) and tectonic tremor (TT) have been observed. At a distance of 300 km from the trench (beneath central Mexico), the plate dives into the mantle with an angle of 76º to a depth of 500 km. This geometry changes abruptly to the south, marking the eastern limit of the USL. This change seems to be also characterized by a tear on the slab. Finally to the south, the Cocos plate subducts with a constant angle of 26º. This presentation summarizes the work of many contributors including A. Arciniega-Ceballos, M. Brudzinski, E. Cabral-Cano, T. Chen, R. Clayton,F. Cordoba-Montiel,P. Davis,S. Dougherty,F. Green, M. Gurnis, D. V. Helmberger, A. Husker,A. Iglesias, Y. Kim, V. Manea, D. Melgar, M. Rodríguez-Domínguez,S. K. Singh, T.-R. A. Song, C. M. Valdés-González, D. Valencia-Cabrera

  2. 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.

  3. 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.

  4. Establishment of a miniaturized enzyme-linked immunosorbent assay for human transferrin quantification using an intelligent multifunctional analytical plate.

    PubMed

    Spies, Peter; Chen, Guo Jun; Gygax, Daniel

    2008-11-01

    A miniaturized enzyme-linked immunosorbent assay (ELISA) with a reaction volume of 5 microl for human transferrin quantification has successfully been developed using an intelligent multifunctional analytical plate (IMAPlate 5RC96), the first miniature analytical platform capable of manually performing parallel liquid transfer, reaction, and analysis. This is the first article to validate the platform for the ELISA application. The data obtained from the standards in this miniaturized ELISA can well be fitted by a one-site binding reaction mode, the coefficient of variation (CV) of the whole plate for an artificial sample (spiking a known concentration of human transferrin into the assay diluent) is 7.0%, and the mean recovery is between 94 and 114% (n=96), comparable to the values from conventional ELISA in a 96-well format plate. The IMAPlate 5RC96-based miniaturized ELISA not only can reduce sample and reagent consumption to 5% of the conventional ELISA but also can shorten the reaction time. Combined with the advantages brought by miniaturization, the easy-to-handle, parallel, and simultaneous liquid transfer features of the IMAPlate 5RC96 provide a completely new lab tool for manually performing high-throughput ELISA. Our results demonstrate that the IMAPlate 5RC96 is a convenient, robust, high-throughput lab device feasible for miniaturized ELISA in an ordinary laboratory.

  5. [The use of blade plate and dynamic screw plate osteosynthesis].

    PubMed

    Oestern, H J; Gänsslen, A

    2010-02-01

    Osteosynthesis in fracture treatment and in some reconstructive procedures with blade plates or dynamic screw systems was the standard procedure for several decades. In this review, the current options and concepts using blade plate osteosynthesis, stabilization of proximal and distal femur fractures and reconstructive procedures with the dynamic hip screw or the dynamic condylar blade are discussed. On the basis of a literature review, the present indications, results and region-specific complications are reported and discussed.Blade plates are used mainly in the context of reconstructive procedures, as well as in the treatment of pseudoarthroses. The Pauwel procedure in femoral neck non-unions is one of the best known indications. In contrast, the dynamic hip screw is the gold standard for stabilization of femoral neck and most pertrochanteric fractures, whereas the dynamic condylar screw is still an alternative to internal fixators for proximal and distal femoral fracture fixations.

  6. The epiphyseal plate: physiology, anatomy, and trauma.

    PubMed

    von Pfeil, Dirsko J F; DeCamp, Charles E

    2009-08-01

    This article reviews the development of long bones, the microanatomy and physiology of the growth plate, the closure times and contribution of different growth plates to overall growth, and the effect of, and prognosis for, traumatic injuries to the growth plate. Details on surgical treatment of growth plate fractures are beyond the scope of this article.

  7. Plate tectonics of the Mediterranean region.

    PubMed

    McKenzie, D P

    1970-04-18

    The seismicity and fault plane solutions in the Mediterranean area show that two small rapidly moving plates exist in the Eastern Mediterranean, and such plates may be a common feature of contracting ocean basins. The results show that the concepts of plate tectonics apply to instantaneous motions across continental plate boundaries.

  8. 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.

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

    SciTech Connect

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

    1984-02-21

    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. 3 figures, 9 tables.

  10. 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.

  11. Stool Culture

    MedlinePlus

    ... products and services. Advertising & Sponsorship: Policy | Opportunities Stool Culture Share this page: Was this page helpful? Also known as: Bacterial Culture, stool; Feces Culture Formal name: Enteric Pathogens Culture, ...

  12. 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.

  13. Multiple-Dynode-Layer Microchannel Plate

    NASA Technical Reports Server (NTRS)

    Woodgate, Bruce E.

    1990-01-01

    Improved microchannel-plate electron image amplifier made of stack of discrete microchannel-plate layers. New plates easier to manufacture because no need to etch long, narrow holes, to draw and bundle thin glass tubes, or to shear plates to give microchannels curvatures necessary for reduction of undesired emission of ions. Discrete dynode layers stacked with slight offset from layer to layer to form microchannel plate with curved channels. Provides for relatively fast recharging of microchannel dynodes, with consequent enhancement of performance.

  14. 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.

  15. Riveted joints in thin plates

    NASA Technical Reports Server (NTRS)

    Hilbes, W

    1930-01-01

    The method of riveting by forming the closing head under increasing pressure or by a series of hammer blows is investigated. The question as to the best edge distance, i.e. that distance from the rivet center to the plate edge below which it is unadvisable to go and, at the same time useless to go beyond, was examined. The slippage of plates which occurs under the effect of a certain stress, does not completely disappear on unloading. The elastic form change is essentially a function of the elasticity modulus, the slip modulus, the supporting moment, the rivet spacing, etc.

  16. License Plate Recognition System for Indian Vehicles

    NASA Astrophysics Data System (ADS)

    Sanap, P. R.; Narote, S. P.

    2010-11-01

    We consider the task of recognition of Indian vehicle number plates (also called license plates or registration plates in other countries). A system for Indian number plate recognition must cope with wide variations in the appearance of the plates. Each state uses its own range of designs with font variations between the designs. Also, vehicle owners may place the plates inside glass covered frames or use plates made of nonstandard materials. These issues compound the complexity of automatic number plate recognition, making existing approaches inadequate. We have developed a system that incorporates a novel combination of image processing and artificial neural network technologies to successfully locate and read Indian vehicle number plates in digital images. Commercial application of the system is envisaged.

  17. 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.

  18. Effects of a sliding plate on morphology of the epiphyseal plate in goat distal femur.

    PubMed

    Lin, Da-sheng; Lian, Ke-jian; Hong, Jia-yuan; Ding, Zhen-qi; Zhai, Wen-liang

    2012-01-01

    The aim of this study was to observe the effects of a sliding plate on the morphology of the epiphyseal plate in goat distal femur. Eighteen premature female goats were divided randomly into sliding plate, regular plate and control groups. Radiographic analysis and histological staining were performed to evaluate the development of epiphyseal plate at 4 and 8 weeks after surgery. In the sliding plate group, the plate extended accordingly as the epiphyseal plate grows, and the epiphyseal morphology was kept essential normal. However, the phenomenon of the epiphyseal growth retardation and premature closure were very common in the regular plate group. In addition, the sliding plate group exhibited more normal histologic features and Safranin O staining compared to the regular plate group. Our results suggest that the sliding plate can provide reliable internal fixation of epiphyseal fracture without inhibiting epiphyseal growth.

  19. Effects of a Sliding Plate on Morphology of the Epiphyseal Plate in Goat Distal Femur

    PubMed Central

    LIN, Da-sheng; LIAN, Ke-jian; HONG, Jia-yuan; DING, Zhen-qi; ZHAI, Wen-liang

    2012-01-01

    The aim of this study was to observe the effects of a sliding plate on the morphology of the epiphyseal plate in goat distal femur. Eighteen premature female goats were divided randomly into sliding plate, regular plate and control groups. Radiographic analysis and histological staining were performed to evaluate the development of epiphyseal plate at 4 and 8 weeks after surgery. In the sliding plate group, the plate extended accordingly as the epiphyseal plate grows, and the epiphyseal morphology was kept essential normal. However, the phenomenon of the epiphyseal growth retardation and premature closure were very common in the regular plate group. In addition, the sliding plate group exhibited more normal histologic features and Safranin O staining compared to the regular plate group. Our results suggest that the sliding plate can provide reliable internal fixation of epiphyseal fracture without inhibiting epiphyseal growth. PMID:22359485

  20. MyPlate Food Guide

    MedlinePlus

    ... whole-wheat bread, brown rice, or oatmeal. 4. Protein Foods that are high in protein help the body build, maintain, and repair tissue. ... on the MyPlate graphic shows the proportion of protein you need. Foods high in protein include beef, ...

  1. Shaker slip-plate adapter

    NASA Technical Reports Server (NTRS)

    Holm, O. S.

    1969-01-01

    Magnesium adapter ties in all of the attachment bosses on a horizontal slip table and makes a rigid coupling which terminates in a single row of attachment bosses at the edge of the horizontal plate. This eliminates ineffective dissipation of the driving force in vibration tests.

  2. Double-Plate Penetration Equations

    NASA Technical Reports Server (NTRS)

    Hayashida, K. B.; Robinson, J. H.

    2000-01-01

    This report compares seven double-plate penetration predictor equations for accuracy and effectiveness of a shield design. Three of the seven are the Johnson Space Center original, modified, and new Cour-Palais equations. The other four are the Nysmith, Lundeberg-Stern-Bristow, Burch, and Wilkinson equations. These equations, except the Wilkinson equation, were derived from test results, with the velocities ranging up to 8 km/sec. Spreadsheet software calculated the projectile diameters for various velocities for the different equations. The results were plotted on projectile diameter versus velocity graphs for the expected orbital debris impact velocities ranging from 2 to 15 km/sec. The new Cour-Palais double-plate penetration equation was compared to the modified Cour-Palais single-plate penetration equation. Then the predictions from each of the seven double-plate penetration equations were compared to each other for a chosen shield design. Finally, these results from the equations were compared with test results performed at the NASA Marshall Space Flight Center. Because the different equations predict a wide range of projectile diameters at any given velocity, it is very difficult to choose the "right" prediction equation for shield configurations other than those exactly used in the equations' development. Although developed for various materials, the penetration equations alone cannot be relied upon to accurately predict the effectiveness of a shield without using hypervelocity impact tests to verify the design.

  3. Comment on "Intermittent plate tectonics?".

    PubMed

    Korenaga, Jun

    2008-06-06

    Silver and Behn (Reports, 4 January 2008, p. 85) proposed that intermittent plate tectonics may resolve a long-standing paradox in Earth's thermal evolution. However, their analysis misses one important term, which subsequently brings their main conclusion into question. In addition, the Phanerozoic eustasy record indicates that the claimed effect of intermittency is probably weak.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Measuring oxygen levels in Caco-2 cultures

    PubMed Central

    Zeitouni, Nathalie E; Fandrey, Joachim; Naim, Hassan Y; von Köckritz-Blickwede, Maren

    2015-01-01

    Purpose Measuring oxygen levels in three different systems of Caco-2 cell culture. Methods Caco-2 cells were cultured in three different systems, using conventional polystyrene 24-well plates, special 24-well gas permeable plates, or on membrane inserts in conventional plates. Optical sensor spots were used to measure dissolved O2 levels in these cultured cells over the course of 6 days under normoxia (143 mmHg) and for 6 hours under hypoxia (7 mmHg). Western blot analysis was used to determine the protein levels of hypoxia-inducible factor 1α (HIF-1α) in the different cultures. Results All culture systems displayed lower O2 levels over time than expected when cultured under normoxia conditions. On average, O2 levels reached as low as 25 mmHg in 24-well plates but remained at 97 and 117 mmHg in gas permeable plates and membrane inserts, respectively. Under hypoxia, 1 mL cell cultures equilibrated to 7 mmHg O2 within the first 60 minutes and dropped to 0.39 and 0.61 mmHg O2 in 24-well and gas permeable plates, respectively, after the 6-hour incubation period. Cultures in membrane inserts did not equilibrate to 7 mmHg by the end of the 6-hour incubation period, where the lowest O2 measurements reached 23.12 mmHg. Western blots of HIF-1α protein level in the whole cell lysates of the different Caco-2 cultures revealed distinct stabilization of HIF-1α after hypoxic incubation for 1, 2, and 4 hours in 24-well plates as well as gas permeable plates. For membrane inserts, notable HIF-1α was seen after 4 hours of hypoxic incubation. Conclusion Cellular oxygen depletion was achieved in different hypoxic Caco-2 culture systems. However, different oxygen levels comparing different culture systems indicate that O2 level should be carefully considered in oxygen-dependent experiments. PMID:27774482

  9. 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...

  10. Direct detection of yeast mutants with reduced viability on plates by erythrosine B staining.

    PubMed

    Bonneu, M; Crouzet, M; Urdaci, M; Aigle, M

    1991-03-02

    In this paper we report a rapid method to screen yeast mutants exhibiting reduced viability directly on plates. This method avoids the need for replica plating and is based on the addition of the vital dye erythrosine B in nutrient medium. After 2 or 3 days of culture, colonies containing a large proportion of dead cells show a pink or a dark pink color whereas normal colonies are practically white.

  11. Plating Processes Utilizing High Intensity Acoustic Beams

    NASA Technical Reports Server (NTRS)

    Oeftering, Richard C. (Inventor); Denofrio, Charles (Inventor)

    2002-01-01

    A system and a method for selective plating processes are disclosed which use directed beams of high intensity acoustic waves to create non-linear effects that alter and improve the plating process. The directed beams are focused on the surface of an object, which in one embodiment is immersed in a plating solution, and in another embodiment is suspended above a plating solution. The plating processes provide precise control of the thickness of the layers of the plating, while at the same time, in at least some incidents, eliminates the need for masking.

  12. 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.

  13. Essential oil of tarragon (Artemisia dracunculus) antibacterial activity on Staphylococcus aureus and Escherichia coli in culture media and Iranian white cheese

    PubMed Central

    Raeisi, M; Tajik, H; Razavi, Roohani SM; Maham, M; Moradi, M; Hajimohammadi, B; Naghili, H; Hashemi, M; Mehdizadeh, T

    2012-01-01

    Background and Objectives In this study, the antibacterial effect of essential oil of tarragon (Artemisia dracunculus) on Staphylococcus aureus and Escherichia coli was evaluated in culture media and Iranian white cheese. Materials and Methods The tarragon essential oil (EO) obtained by the steam distillation method and its antibacterial activity was evaluated in 96-well microtiter plates containing brain heart infusion broth. The enumeration of S. aureus and E. coli in cheese samples were carried out on the following media: Baired parker agar for S.aureus, incubated at 37°C for 24 h; and MacConkey sorbitol agar for E. coli, incubated at 37°C for 24 h. Iranian white cheese was produced from fresh and whole pasteurized cow milk (2.5%). Bacteria (103 cfu/mL) were inoculated to different batches. Cheese was treated with different concentrations of EO (15 and 1500 µg/mL) and separated into four parts in an equal manner. The sensory evaluation was done by a panel of four judges. Results According to the results obtained, minimum inhibitory concentrations (MIC) for E. coli and S. aureus were 2500 and 1250 µg/mL, respectively. Also, minimum bactericidal concentration (MBC) for the mentioned microorganisms were 5000 and 2500 µg/mL, respectively. All the EO concentrations for each bacteria result in reducing bacterial count of cheese samples compared to control (P<0.05). Also, with increasing concentration of EO in cheese samples, the bacterial count was reduced further (P<0.05). Conclusion Based on our findings, tarragon essential oil has antibacterial effect on two important pathogen bacteria (S. aureus and E. coli) and can be applied as a preservative in foods such as cheese. PMID:22783458

  14. 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.

  15. Absorbable plate strength loss during molding.

    PubMed

    Ballard, Tiffany N S; Kelly, Kevin J; Zaydfudim, Victor; Walcutt, Noah L; Lahijani, Soheil S; Shack, R Bruce; Thayer, Wesley P

    2010-05-01

    Bioabsorbable plating systems play an integral role in cranial vault remodeling. After experiencing a case of plate failure requiring emergent reexploration, we investigated the potential causes. We hypothesize that extended submersion in the molding bath during plate preparation might advance the rate of hydrolysis and compromise plate structural integrity. Using an absorbable poly-D/L-lactic acid plating system, we assessed the effect of extended submersion on plate strength and stiffness when loaded in a cantilever fashion and with pure tension. We assessed these differences with the Student t test and linear regression modeling. We also generated a computer model of the plates for finite element analysis. When left in the molding bath for extended periods, the plates changed color and lost strength. After 5 minutes, 30% of maximum plate load capacity was lost in a cantilever beam test (P < 0.001) consistent with use of a 15% thinner plate. Tensile testing revealed the initial elastic modulus of 6.42 +/- 0.13 GPa decreased 16% to 5.41 +/- 0.50 GPa after 5 minutes of submersion (P = 0.027). The changes in plate strength and elastic modulus both worsened with increased submersion times. Finite element analysis of the plates also predicted clinically significant increases in plate deviation under normal loading conditions. Our study demonstrates that extended submersion of absorbable plates during molding results in a significant loss of plate strength and stiffness. Further, our computer model predicts that these changes could result in an unacceptable plate deviation under normal loading conditions. Together, these data caution against overmolding of plates to avoid compromising their structural integrity.

  16. Primary plate identification of group A beta-hemolytic streptococci utilizing a two-disk technique.

    PubMed Central

    Baron, E J; Gates, J W

    1979-01-01

    A two-disk system is described which allows primary plate identification of group A beta-hemolytic streptococci. Group A beta-hemolytic streptococci could be visualized on primary throat culture plates by using trimethoprim-sulfamethoxazole to inhibit normal flora. In the heavily inoculated area of Trypticase soy agar plates containing 5% sheep blood, a 25-microgram/ml trimethoprim-sulfamethoxazole disk was placed contiguous to a 0.04-U bacitracin disk. A total of 259 throat specimens were examined with this two-disk system. The swabs from these throat specimens were incubated in Todd-Hewitt broth. The bacterial pellet from the broths was stained by fluorescent antibody as a control. Of the cultures that were determined to be positive on the plates, 75% could be read unequivocally after overnight incubation, whereas the remaining 25% required subculture. The plates recovered 91% of the cultures which were considered as true positives by the broth-fluorescent-antibody technique. This method provided a significant savings in time compared with standard plate methods and in cost of materials compared with broth-fluorescent-antibody methods. This technique is particularly valuable for producing rapid results in laboratories where fluorescence microscopy would not be cost-effective. Images PMID:387811

  17. Plate Boundary Observatory in Taiwan

    NASA Astrophysics Data System (ADS)

    Yu, S.; Tsai, C.

    2003-12-01

    The island of Taiwan is situated in the plate boundary zone between the Eurasian and the Philippine Sea plates. The Philippine Sea plate is subducting northwestward underneath the Eurasian plate along the Ryukyu Trench in the north, while the Eurasian plate underthrusts the Philippine Sea plate along the Manila Trench in the south. Taking advantage of the extremely high strain rate in the Taiwan area, an integrated National Science Council project, Plate Boundary Observatory in Taiwan (PBOT), was initiated following the idea of US PBO. The scientific goal of PBOT is to observe the crustal deformation on various temporal and spatial scales in the Taiwan plate boundary zone employing available state of the art techniques for measuring crustal strain. The techniques include seismology, Global Positioning System (GPS), Interferometric Synthetic Aperture Radar (InSAR), borehole strainmeter, and earthquake geology. They are complementary to each other and form a complete spectrum of measuring various periods of crustal strain. The process of crustal deformation is generally quite slow. To obtain a reliable result, we usually need to persist in the observations for several years or even decades. Thus the PBOT should be a long-term project. In the first phase of 3 years period from 2003 to 2006, we will focus on the two areas, i.e. the plate suture zone in the Longitudinal Valley area and the western Taiwan where the higher seismic hazard is expected. A five-year national program, entitled ­Program for Earthquake and Active-fault Research (PEAR)­" was initiated after the disastrous 1999 Chi-Chi earthquake (Mw 7.6). As part of the PEAR, a dense continuous GPS array consisting of 150 new and about 50 pre-existing stations will be completed in the Taiwan area by the end of 2005 through a joint effort by the Central Weather Bureau and the Institute of Earth Sciences, Academia Sinica. The 50 new stations are going to be evenly distributed around the Taiwan Island. The other

  18. Subduction Drive of Plate Tectonics

    NASA Astrophysics Data System (ADS)

    Hamilton, W. B.

    2003-12-01

    Don Anderson emphasizes that plate tectonics is self-organizing and is driven by subduction, which rights the density inversion generated as oceanic lithosphere forms by cooling of asthenosphere from the top. The following synthesis owes much to many discussions with him. Hinge rollback is the key to kinematics, and, like the rest of actual plate behavior, is incompatible with bottom-up convection drive. Subduction hinges (which are under, not in front of, thin leading parts of arcs and overriding plates) roll back into subducting plates. The Pacific shrinks because bounding hinges roll back into it. Colliding arcs, increasing arc curvatures, back-arc spreading, and advance of small arcs into large plates also require rollback. Forearcs of overriding plates commonly bear basins which preclude shortening of thin plate fronts throughout periods recorded by basin strata (100 Ma for Cretaceous and Paleogene California). This requires subequal rates of advance and rollback, and control of both by subduction. Convergence rate is equal to rates of rollback and advance in many systems but is greater in others. Plate-related circulation probably is closed above 650 km. Despite the popularity of concepts of plumes from, and subduction into, lower mantle, there is no convincing evidence for, and much evidence against, penetration of the 650 in either direction. That barrier not only has a crossing-inhibiting negative Clapeyron slope but also is a compositional boundary between fractionated (not "primitive"), sluggish lower mantle and fertile, mobile upper mantle. Slabs sink more steeply than they dip. Slabs older than about 60 Ma when their subduction began sink to, and lie down on and depress, the 650-km discontinuity, and are overpassed, whereas younger slabs become neutrally buoyant in mid-upper mantle, into which they are mixed as they too are overpassed. Broadside-sinking old slabs push all upper mantle, from base of oceanic lithosphere down to the 650, back under

  19. Effective characteristics of corrugated plates

    NASA Astrophysics Data System (ADS)

    Arkhangel'skii, A. F.; Gorbachev, V. I.

    2007-06-01

    Corrugated plates are widely used in modern constructions and structures, because they, in contrast to plane plates, possess greater rigidity. In many cases, such a plate can be modeled by a homogeneous anisotropic plate with certain effective flexural and tensional rigidities. Depending on the geometry of corrugations and their location, the equivalent homogeneous plate can also have rigidities of mutual influence. These rigidities allow one to take into account the influence of bending moments on the strain in the midplane and, conversely, the influence of longitudinal strains on the plate bending [1]. The behavior of the corrugated plate under the action of a load normal to the midsurface is described by equations of the theory of flexible plates with initial deflection. These equations form a coupled system of nonlinear partial differential equations with variable coefficients [2]. The dependence of the coefficients on the coordinates is determined by the corrugation geometry. In the case of a plate with periodic corrugation, the coefficients significantly vary within one typical element and depend on the values of local variables determined in each of the typical elements. There is a connection between the local and global variables, and therefore, the functions of local coordinates are simultaneously functions of global coordinates, which are sometimes called rapidly oscillating functions [3]. One of the methods for solving the equations with rapidly oscillating coefficients is the asymptotic method of small geometric parameter. The standard procedure of this method usually includes preparatory stages. At the first stage, as a rule, a rectangular periodicity cell is distinguished to be a typical element. At the second stage, the scale of global coordinates is changed so that the rectangular structure periodicity cells became square cells of size l × l. The third stage consists in passing to the dimensionless global coordinates relative to the plate

  20. Rapid screening of purification strategies for the capture of a human recombinant F(ab')2 expressed in baculovirus-infected cells using a micro-plate approach and SELDI-MS.

    PubMed

    Pezzini, J; Brenac Brochier, V; Barrouillet, M P; Cerruti, M; Clofent-Sanchez, G; Schapman, A; Topol, A; Robert, R; Cabanne, C; Cerruti, P; Santarelli, X

    2009-08-15

    The development of a capture step of a human recombinant F(ab')(2) produced and expressed in baculovirus-infected cells was investigated by screening three mixed-mode chromatography sorbents (HEA HyperCel, PPA HyperCel and MEP HyperCel) and two ion exchangers (Q Ceramic HyperD F, S Ceramic HyperD F) sorbents using a 96-well plate format and SELDI-MS. HEA HyperCel gave the best separation performance therefore the conditions tested in micro-plate were transferred to laboratory scale chromatographic experiments, confirming that the recombinant F(ab')(2) was effectively captured on the mixed-mode sorbent without any pre-treatment of the crude extract with a 82% recovery and a 39-fold purification.

  1. High thermal capacity cold plate/hot plate

    NASA Technical Reports Server (NTRS)

    Hwangbo, H.; Mcever, W. S.

    1985-01-01

    The results of an analytical study to determine the feasibility of a novel two-phase cold plate/hot plate (CPHP) are presented. A key feature of the CPHP is the use of capillary forces to separate the liquid and vapor phases and distribute the liquid over the evaporating/condensing surface. The liquid phase is held by capillary forces in a reservoir and is carried to the evaporating surface by a wick. The reservoir is replenished at intervals from a valved external liquid supply line. In the hot plate mode, liquid accumulates in the reservoir and is removed by an external condensate line. Performance requirements for the device were capability of handling a power density of 4 watts/sq cm, an outlet quality (percentage of vapor flow to total flow) greater than 90 percent, and operation in a 0-g environment. An analytical model of CPHP operation was developed which concentrated on the liquid and vapor flows. It was found that the liquid carrying capillary grooves of rectangular cross-section gave significantly better predicted performance than V-shaped grooves. Using the analytical model, capillary groove width and length, vapor channel dimensions, and other parameters were selected to meet the performance requirements with a 100-percent margin.

  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. Urine Culture

    MedlinePlus

    ... products and services. Advertising & Sponsorship: Policy | Opportunities Urine Culture Share this page: Was this page helpful? Also known as: Urine Culture and Sensitivity; Urine C and S Formal name: Culture, ...

  4. 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)

  5. 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...

  6. 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...

  7. 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...

  8. 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...

  9. 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...

  10. 30 CFR 20.13 - Approval plate.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... manufacturer shall attach, stamp, or mold an approval plate on the battery container or housing of each... has been adjudged safe for use in gassy and dusty mines. (b) Use of approval plate. Permission...

  11. 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.

  12. Expansion of adipose mesenchymal stromal cells is affected by human platelet lysate and plating density.

    PubMed

    Cholewa, Dominik; Stiehl, Thomas; Schellenberg, Anne; Bokermann, Gudrun; Joussen, Sylvia; Koch, Carmen; Walenda, Thomas; Pallua, Norbert; Marciniak-Czochra, Anna; Suschek, Christoph V; Wagner, Wolfgang

    2011-01-01

    The composition of mesenchymal stromal cells (MSCs) changes in the course of in vitro culture expansion. Little is known how these cell preparations are influenced by culture media, plating density, or passaging. In this study, we have isolated MSCs from human adipose tissue in culture medium supplemented with either fetal calf serum (FCS) or human platelet lysate (HPL). In addition, culture expansion was simultaneously performed at plating densities of 10 or 10,000 cells/cm(2). The use of FCS resulted in larger cells, whereas HPL significantly enhanced proliferation. Notably, HPL also facilitated expansion for more population doublings than FCS (43 ± 3 vs. 22 ± 4 population doubling; p < 0.001), while plating density did not have a significant effect on long-term growth curves. To gain further insight into population dynamics, we conceived a cellular automaton model to simulate expansion of MSCS. It is based on the assumptions that the number of cell divisions is limited and that due to contact inhibition proliferation occurs only at the rim of colonies. The model predicts that low plating densities result in more heterogeneity with regard to cell division history, and favor subpopulations of higher migratory activity. In summary, HPL is a suitable serum supplement for isolation of MSC from adipose tissue and facilitates more population doublings than FCS. Cellular automaton computer simulations provided additional insights into how complex population dynamics during long-term expansion are affected by plating density and migration.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. Optical Near-Field Plates

    DTIC Science & Technology

    2015-04-08

    microscopy images of seven plasmonic color filters illuminated by the white microscope light. Scale bar is 10 µm. (b) Experimentally measured transmission... color filtering and spectral imaging ,” Nat. Comm. 1, 59 (2010). 3. H.-F. Shi and L. J. Guo, “Design of Plasmonic Near Field Plate at Opitical...wavelength. Plasmonic filters were developed and transmission color filtering was accomplished. The polarization property and the pitch-dependent

  2. 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

  3. Optimization of neutron imaging plate

    NASA Astrophysics Data System (ADS)

    Haga, Y. K.; Neriishi, K.; Takahashi, K.; Niimura, N.

    2002-07-01

    Considering the elementary processes of neutron detection occurring in the neutron imaging plate (NIP) has optimized the performance of NIP. For these processes, the color center creation efficiencies ( ɛcc values) have been experimentally determined with NIPs which have different mole fraction of photostimulated (PSL) material ( φPSL values) and different thickness ( t). The effectiveness of the optimization procedure has been demonstrated by the measurement of the neutron diffraction intensities from a hen egg-white lysozyme protein crystal.

  4. Plating Waste Sludge Metal Recovery.

    DTIC Science & Technology

    1985-05-30

    l - ~ ’.4 41 *4 7.1 4...34 L €5 ’ ’ . ’ " " r ,*: -: ,., -. . .,,:€ ,, .. € : ....- -1. PHASE I - UERATURE REVIEW Phase I of the project "Plating Waste Sludge Metal Recovery...8217’’,. ’.’.."..-’......’ ...’ , m .m llll- l ..m~ l : j nm~, l ~linn~ . l , b~~h~h , . r - r. -*1 . * , * . . , .d PHASE IV -

  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. 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.

  7. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 21 CFR 872.4760 - Bone plate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 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...

  14. 21 CFR 872.4760 - Bone plate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 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...

  15. 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...

  16. 21 CFR 872.4760 - Bone plate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 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...

  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. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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)

  3. Crustal genesis and plate tectonics

    NASA Astrophysics Data System (ADS)

    Yang-shen, Shi; Shu-feng, Yang; Ling-zhi, Guo; Huo-gen, Dong

    1991-02-01

    As a transitional zone between ocean and continent, the continental margins are important accreted belts of the granitic crust. Extensive granitoids of different geological ages and different genetic series occur in the continental margins of southeastern China. The granites in active continental margins of southeast China are divided into two genetic series by present authors as follows:(1) S-type granitoids in the interior of the continental plate; and (2) I-type granitoids in the margin of the continental plate. S-type and I-type granitoids of almost the same geological age are paired and belted in their spatial distribution parallel to the continental margin. These are referred to as "paired granite belts". Granitic continental crust genesis is closely related to the growth of paired granite belts. In Mt. Yunkai of southeastern China, Guangdong province, Early Paleozoic S-type granitoids have been produced as a result of transformation of Cambrian sediments during the processes of metamorphism, migmatization, granitization and remelting. Co-magmatic volcanic rocks are completely lacking in this area. Early Paleozoic I-type granitoids on the east side of Mt. Yunkai are distributed along the ancient continental margin. These rocks co-exist with comagmatic island-arc volcanic rocks of calc-alkaline series in this area. The above mentioned two types of granitoids are paired and belted. Similar paired granite belts exist around the world, such as in North America and southeastern Australia, and are interpreted to be products of post plate convergence, subduction and collision.

  4. A simple micro-extraction plate assay for automated LC-MS/MS analysis of human serum 25-hydroxyvitamin D levels.

    PubMed

    Geib, Timon; Meier, Florian; Schorr, Pascal; Lammert, Frank; Stokes, Caroline S; Volmer, Dietrich A

    2015-01-01

    This short application note describes a simple and automated assay for determination of 25-hydroxyvitamin D (25(OH)D) levels in very small volumes of human serum. It utilizes commercial 96-well micro-extraction plates with commercial 25(OH)D isotope calibration and quality control kits. Separation was achieved using a pentafluorophenyl liquid chromatography column followed by multiple reaction monitoring-based quantification on an electrospray triple quadrupole mass spectrometer. Emphasis was placed on providing a simple assay that can be rapidly established in non-specialized laboratories within days, without the need for laborious and time consuming sample preparation steps, advanced calibration or data acquisition routines. The analytical figures of merit obtained from this assay compared well to established assays. To demonstrate the applicability, the assay was applied to analysis of serum samples from patients with chronic liver diseases and compared to results from a routine clinical immunoassay.

  5. 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.

  6. 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

  7. A Simplified Culture Method for Specific Diagnosis of Cerebrospinal Meningitis.

    DTIC Science & Technology

    efficient than chocolate agar and blood agar plates when tested in parallel on 460 cerebrospinal fluid specimens. The culture bottles are inexpensive, portable, and durable making them highly suited to studies in the field. (Author)

  8. 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

  9. 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.

  10. Wheelspace windage cover plate for turbine

    DOEpatents

    Lathrop, Norman Douglas

    2002-01-01

    Windage cover plates are secured between the wheels and spacer of a turbine rotor to prevent hot flow path gas ingestion into the wheelspace cavities. Each cover plate includes a linear, axially extending body curved circumferentially with a radially outwardly directed wall at one axial end. The wall defines a axially opening recess for receiving a dovetail lug. The cover plate includes an axially extending tongue received in a circumferential groove of the spacer. The cover plate is secured with the tongue in the groove and dovetail lug in the recess. Lap joints between circumferentially adjacent cover plates are provided.

  11. Induction and characterization of metallothionein in chicken epiphyseal growth plate cartilage chondrocytes.

    PubMed

    Sauer, G R; Nie, D; Wu, L N; Wuthier, R E

    1998-01-01

    Following exposure to cadmium or zinc, chickens were sacrificed and the liver, kidney, and bone epiphyseal growth plates harvested. When cytosolic extracts of the growth plate cartilage were fractionated by gel filtration chromatography, a protein with high metal-binding capacity and low ultraviolet (UV) absorbance eluted in the same position as liver metallothionein (MT) and a MT standard. Cd or Zn treatment resulted in a 25-fold or 5-fold induction in growth plate MT, respectively. In liver the greatest level of MT induction was seen with short-term Cd exposures. In contrast, MT levels in the growth plate increased as the duration of Cd exposure increased. Induction of MT in growth plate chondrocyte cell cultures was observed for media Cd concentrations of > or = 0.1 microM and Zn concentrations of > or = 100 microM. Basal and inducible levels of MT declined through the culture period and were lowest in the terminally differentiated mineralized late stages of the culture. Alkaline phosphatase activity was also lowest in the late-stage cultures, while total cellular protein increased throughout the culture period. Treatment of chondrocytes with Zn prior to Cd exposure resulted in a protective induction of MT. Pre-treatment of chondrocytes with dexamethasone resulted in suppressed synthesis of MT upon Cd exposure and greater Cd toxicity. Both Cd and Zn resulted in significantly increased levels of MT mRNA in chondrocyte cell cultures. Dexamethasone treatment resulted in an approximate 2- to 3-fold increase in MT mRNA. This is contrary to the finding that MT protein levels were decreased by dexamethasone. The findings suggest that an increased rate of MT degradation in dexamethasone-treated and late-stage chondrocyte cultures may be associated with the terminally differentiated phenotype.

  12. Development of flat - plate solar plate collector - evaporator. Summary report

    SciTech Connect

    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.

  13. 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.

  14. Absolute Plate Velocities from Seismic Anisotropy

    NASA Astrophysics Data System (ADS)

    Kreemer, Corné; Zheng, Lin; Gordon, Richard

    2015-04-01

    The orientation of seismic anisotropy inferred beneath plate interiors may provide a means to estimate the motions of the plate relative to the sub-asthenospheric mantle. Here we analyze two global sets of shear-wave splitting data, that of Kreemer [2009] and an updated and expanded data set, to estimate plate motions and to better understand the dispersion of the data, correlations in the errors, and their relation to plate speed. We also explore the effect of using geologically current plate velocities (i.e., the MORVEL set of angular velocities [DeMets et al. 2010]) compared with geodetically current plate velocities (i.e., the GSRM v1.2 angular velocities [Kreemer et al. 2014]). We demonstrate that the errors in plate motion azimuths inferred from shear-wave splitting beneath any one tectonic plate are correlated with the errors of other azimuths from the same plate. To account for these correlations, we adopt a two-tier analysis: First, find the pole of rotation and confidence limits for each plate individually. Second, solve for the best fit to these poles while constraining relative plate angular velocities to consistency with the MORVEL relative plate angular velocities. The SKS-MORVEL absolute plate angular velocities (based on the Kreemer [2009] data set) are determined from the poles from eight plates weighted proportionally to the root-mean-square velocity of each plate. SKS-MORVEL indicates that eight plates (Amur, Antarctica, Caribbean, Eurasia, Lwandle, Somalia, Sundaland, and Yangtze) have angular velocities that differ insignificantly from zero. The net rotation of the lithosphere is 0.25±0.11° Ma-1 (95% confidence limits) right-handed about 57.1°S, 68.6°E. The within-plate dispersion of seismic anisotropy for oceanic lithosphere (σ=19.2° ) differs insignificantly from that for continental lithosphere (σ=21.6° ). The between-plate dispersion, however, is significantly smaller for oceanic lithosphere (σ=7.4° ) than for continental

  15. 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.

  16. 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.

  17. 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

  18. Anti and Androgenic Activities in MDA-KB2 Cells: A Comparison of Performance in 96 Well Versus HTS Assays

    EPA Science Inventory

    We developed the MDA-kb2 cell line to screen androgen agonists/antagonists (Wilson et al., ToxSci 66:69, 2002). MDA-kb2 has been used to quantify anti- and androgenic activities of chemicals, mixtures, combustion by-products, oil dispersants and waste, source and drinking water s...

  19. High-throughput synthesis of conopeptides: a safety-catch linker approach enabling disulfide formation in 96-well format.

    PubMed

    Brust, Andreas; Tickle, Alice E

    2007-02-01

    Conotoxins exhibit a high degree of selectivity and potency for a range of pharmacologically relevant targets. The rapid access to libraries of conotoxin analogues, containing multiple intramolecular disulfide bridges for use in drug development, can be a very labor intensive, multi-step task. This work describes a high-throughput method for the synthesis of cystine-bridged conopeptides. Peptides were assembled on a peptide synthesizer employing the Fmoc solid-phase strategy using a safety-catch amide linker (SCAL). Side-chain protecting groups were removed on solid phase before SCAL activation with ammonium iodide in TFA, finally releasing the peptide into the TFA solution. Disulfide bond formation was performed in the cleavage mixture employing DMSO. This improved method allows mixtures of oxidized peptides to be obtained in parallel directly from a peptide synthesizer. A single HPLC purification of the resulting crude oxidized material produced peptides of > 95% purity.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. Development of an automated MODS plate reader to detect early growth of Mycobacterium tuberculosis.

    PubMed

    Comina, G; Mendoza, D; Velazco, A; Coronel, J; Sheen, P; Gilman, R H; Moore, D A J; Zimic, M

    2011-06-01

    In this work, an automated microscopic observation drug susceptibility (MODS) plate reader has been developed. The reader automatically handles MODS plates and after autofocussing digital images are acquired of the characteristic microscopic cording structures of Mycobacterium tuberculosis, which are the identification method utilized in the MODS technique to detect tuberculosis and multidrug resistant tuberculosis. In conventional MODS, trained technicians manually move the MODS plate on the stage of an inverted microscope while trying to locate and focus upon the characteristic microscopic cording colonies. In centres with high tuberculosis diagnostic demand, sufficient time may not be available to adequately examine all cultures. An automated reader would reduce labour time and the handling of M. tuberculosis cultures by laboratory personnel. Two hundred MODS culture images (100 from tuberculosis positive and 100 from tuberculosis negative sputum samples confirmed by a standard MODS reading using a commercial microscope) were acquired randomly using the automated MODS plate reader. A specialist analysed these digital images with the help of a personal computer and designated them as M. tuberculosis present or absent. The specialist considered four images insufficiently clear to permit a definitive reading. The readings from the 196 valid images resulted in a 100% agreement with the conventional nonautomated standard reading. The automated MODS plate reader combined with open-source MODS pattern recognition software provides a novel platform for high throughput automated tuberculosis diagnosis.

  5. Yield Surfaces for Anisotropic Plates

    NASA Astrophysics Data System (ADS)

    Walker, J. D.; Thacker, B. H.

    1999-06-01

    Modern aerospace systems are incorporating composite materials into their structures. Often, the composite materials are anisotropic in their mechanical response due to the geometric layout of fibers. For many years, the failure surfaces of anisotropic materials were thought to be characterizable by a quadratic function in the stress, often referred to as a Tsai-Wu yield surface, or, in a more restrictive form, a Tsai-Hill yield surface. Such a representation does not work for materials that are strong in two directions and weak in one direction, which, unfortunately, is the case of most interest since it represents most composite plates. This paper demonstrates the impossibility of modeling the failure surface with both the Tsai-Wu and Tsai-Hill failure surfaces. We then present a yield surface based on the lemniscate, which is quartic in the stress. This new yield surface addresses the case of strong in two directions and weak in one. Calculations with a fragment impacting a composite plate modeled with the new yield surface are presented. Modifications of the yield surface are presented to allow, in a limited way, materials that are both anisotropic and have differing strengths in tension and compression.

  6. Silicon-micromachined microchannel plates

    NASA Astrophysics Data System (ADS)

    Beetz, Charles P.; Boerstler, Robert; Steinbeck, John; Lemieux, Bryan; Winn, David R.

    2000-03-01

    Microchannel plates (MCP) fabricated from standard silicon wafer substrates using a novel silicon micromachining process, together with standard silicon photolithographic process steps, are described. The resulting SiMCP microchannels have dimensions of ˜0.5 to ˜25 μm, with aspect ratios up to 300, and have the dimensional precision and absence of interstitial defects characteristic of photolithographic processing, compatible with positional matching to silicon electronics readouts. The open channel areal fraction and detection efficiency may exceed 90% on plates up to 300 mm in diameter. The resulting silicon substrates can be converted entirely to amorphous quartz (qMCP). The strip resistance and secondary emission are developed by controlled depositions of thin films, at temperatures up to 1200°C, also compatible with high-temperture brazing, and can be essentially hydrogen, water and radionuclide-free. Novel secondary emitters and cesiated photocathodes can be high-temperature deposited or nucleated in the channels or the first strike surface. Results on resistivity, secondary emission and gain are presented.

  7. 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.

  8. 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.

  9. Principles of cancer cell culture.

    PubMed

    Cree, Ian A

    2011-01-01

    The basics of cell culture are now relatively common, though it was not always so. The pioneers of cell culture would envy our simple access to manufactured plastics, media and equipment for such studies. The prerequisites for cell culture are a well lit and suitably ventilated laboratory with a laminar flow hood (Class II), CO(2) incubator, benchtop centrifuge, microscope, plasticware (flasks and plates) and a supply of media with or without serum supplements. Not only can all of this be ordered easily over the internet, but large numbers of well-characterised cell lines are available from libraries maintained to a very high standard allowing the researcher to commence experiments rapidly and economically. Attention to safety and disposal is important, and maintenance of equipment remains essential. This chapter should enable researchers with little prior knowledge to set up a suitable laboratory to do basic cell culture, but there is still no substitute for experience within an existing well-run laboratory.

  10. 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.

  11. 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).

  12. 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.

  13. Silencing using flexible plate in a duct

    NASA Astrophysics Data System (ADS)

    Ramamoorthy, Sripriya; Grosh, Karl; Nawar, Tony G.

    2002-11-01

    A flexible plate interacting with air in a duct can provide passive means for low frequency broadband transmission loss. The sensitivity of the system to various parameters including unintentionally applied tension, effect of external fluid loading, lateral plate cross modes, structural and acoustic boundary condition are analyzed through experimental measurements and theoretical predictions. In order to avoid breakout noise, a backing cavity can be introduced below the plate. This introduces differences in filtering characteristics. Compared to plate in a single duct, the two-duct system will have higher plate resonance frequencies due to cavity loading on the plate. Means to achieve low frequency broadband transmission loss using two-duct silencers will be discussed. Significance of three dimensionality of the problem will be brought out by comparing the results of three-dimensional finite-element analysis with experimental data. Successful designs and experiments for low frequency attenuation will be presented. [Work supported by NSF and ONR.

  14. 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.

  15. 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).

  16. Flutter of laminated plates in supersonic flow

    NASA Technical Reports Server (NTRS)

    Sawyer, J. W.

    1975-01-01

    A solution procedure was developed using linear small deflection theory for the flutter of simply supported laminated plates. For such plates, the bending and extensional governing equations are coupled and have cross-stiffness terms which do not appear in classical plate theory. An extended Galerkin method is used to obtain approximate solutions to the governing equations, and the aerodynamic pressure loading used in the analysis is that given by linear piston theory with flow at arbitrary cross-flow angle. A limited parametric study was conducted for typical laminated composite plates. The calculations show that both the bending-extensional coupling and the cross-stiffness terms have a large destabilizing effect on flutter. Since classical plate theory does not consider bending-extensional coupling and cross stiffness terms, it usually gives inaccurate and nonconservative flutter boundaries for laminated plates.

  17. 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.

  18. Coupling between plate vibration and acoustic radiation

    NASA Technical Reports Server (NTRS)

    Frendi, Abdelkader; Maestrello, Lucio; Bayliss, Alvin

    1993-01-01

    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.

  19. Reproducibility of Uniform Spheroid Formation in 384-Well Plates: The Effect of Medium Evaporation.

    PubMed

    Das, Viswanath; Fürst, Tomáš; Gurská, Soňa; Džubák, Petr; Hajdúch, Marián

    2016-10-01

    Spheroid cultures of cancer cells reproduce the spatial dimension-induced in vivo tumor traits more effectively than the conventional two-dimensional cell cultures. With growing interest in spheroids for high-throughput screening (HTS) assays, there is an increasing demand for cost-effective miniaturization of reproducible spheroids in microtiter plates (MPs). However, well-to-well variability in spheroid size, shape, and growth is a frequently encountered problem with almost every culture method that has prevented the transfer of spheroids to the HTS platform. This variability partly arises due to increased susceptibility of MPs to edge effects and evaporation-induced changes in the growth of spheroids. In this study, we examined the effect of evaporation on the reproducibility of spheroids of tumor and nontumor cell lines in 384-well plates, and show that culture conditions that prevent evaporation-induced medium loss result in the formation of uniform spheroids across the plate. Additionally, we also present a few technical improvements to increase the scalability of the liquid-overlay spheroid culturing technique in MPs, together with a simple software routine for the quantification of spheroid size. We believe that these cost-effective improvements will aid in further improvement of spheroid cultures for HTS drug discovery.

  20. 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)

  1. Bronchoscopic culture

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/003748.htm Bronchoscopic culture To use the sharing features on this page, please enable JavaScript. Bronchoscopic culture is a laboratory exam to check a piece ...

  2. Throat Culture

    MedlinePlus

    ... products and services. Advertising & Sponsorship: Policy | Opportunities Throat Culture Share this page: Was this page helpful? Collecting | ... treatment | Getting results | see BLOOD SAMPLE Collecting A culture is a test that is often used to ...

  3. Bile culture

    MedlinePlus

    ... these risks. Alternative Names Culture - bile Images Bile culture References Hall GS, Woods GL. Medical bacteriology. In: McPherson RA, Pincus MR, eds. Henry's Clinical Diagnosis and Management by Laboratory Methods . 22nd ed. Philadelphia, PA: Elsevier ...

  4. Esophageal culture

    MedlinePlus

    Culture - esophageal ... There, it is placed in a special dish (culture) and watched for the growth of bacteria, fungi, ... and Fordtran's Gastrointestinal and Liver Disease Pathophysiology/Diagnosis/Management . 10th ed. Philadelphia, PA: Elsevier Saunders; 2016:chap ...

  5. Characterization of Early Cortical Neural Network Development in Multiwell Microelectrode Array Plates

    EPA Science Inventory

    We examined the development of neural network activity using microelectrode array (MEA) recordings made in multi-well MEA plates (mwMEAs) over the first 12 days in vitro (DIV). In primary cortical cultures made from postnatal rats, action potential spiking activity was essentiall...

  6. 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.

  7. Finite Element Anlaysis of Laminated Composite Plates

    DTIC Science & Technology

    1988-09-01

    4.2, results depicting maximum displacement obtained using 2 x 2 integration points, 3 x 3 integration points and ’ heterosis ’ [Ref. 4] elements are...thick and thin plates. This element gives better predictions for thick plates than heterosis ele- ment, however, for thin plates, heterosis element...results showing the normalized maximum displacements are shown in Figure 4.8. The heterosis element results in about ten percent error while the

  8. High performance flat plate solar collector

    NASA Technical Reports Server (NTRS)

    Lansing, F. L.; Reynolds, R.

    1976-01-01

    The potential use of porous construction is presented to achieve efficient heat removal from a power producing solid and is applied to solar air heaters. Analytical solutions are given for the temperature distribution within a gas-cooled porous flat plate having its surface exposed to the sun's energy. The extracted thermal energy is calculated for two different types of plate transparency. Results show the great improvement in performance obtained with porous flat plate collectors as compared with analogous nonporous types.

  9. 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.

  10. Beyond Culture.

    ERIC Educational Resources Information Center

    Barron, Daniel D.

    1993-01-01

    Discusses the lack of literature relating to cultural differences and school library media programs and reviews the book "Beyond Culture" by Edward T. Hall. Highlights include the population/environment crisis, cultural literacy, the use of technology, and Marshall McLuhan's idea of the global village. (LRW)

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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)

  17. 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.

  18. 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.

  19. Yield surfaces for anisotropic plates

    NASA Astrophysics Data System (ADS)

    Walker, J. D.; Thacker, B. H.

    2000-04-01

    Aerospace systems are incorporating composite materials into their structures. The composite materials are often anisotropic in mechanical response due to their geometric layout. For many years, the failure surfaces of anisotropic materials were thought to be characterizable by a quadratic function in the stress, referred to as a Tsai-Wu yield surface, or, in a more restrictive form, a Tsai-Hill yield surface. Such a representation does not work for materials that are strong in two directions and weak in one direction, which is the case of most interest since it represents fiber/epoxy composite plates. This paper demonstrates the impossibility of modeling the failure surface with either the Tsai-Wu or Tsai-Hill failure surfaces. A yield surface is presented based on the lemniscate, which is quartic in the stress. This new yield surface addresses the case of strong in two directions and weak in one.

  20. 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).

  1. Composition of the continental plates

    USGS Publications Warehouse

    Gilluly, J.

    1954-01-01

    The structures of continental plates and of oceanic basins suggested by several seismologists are utilized to estimate the relative volumes of sial and sima in the earth's crust. It seems that sial of the composition of the average igneous rock constitutes fully 26% and perhaps as much as 43% of the total crust. This ratio is far higher than seems likely if the sial had been entirely derived through fractional crystallization of a basaltic magma. The relative paucity of intermediate rocks as compared with granite and gabbro in the crust points in the same direction. The tentative conclusion is reached that the sial owes a large part of its volume to some process other than fractional crystallization of basalt-possibly to the emanation of low-melting constituents such as water, silica, potassa, soda, and alumina directly from the mantle to the crust. ?? 1954 Springer-Verlag.

  2. Inter- and intra-plate deformation at North American plate boundaries

    NASA Technical Reports Server (NTRS)

    Beavan, John

    1986-01-01

    Alaska tectonics and earthquake hazard studies; Southern California tectonics (block rotation); spreading near the Salton Trough; California plate motion (fault zone kinematics); and Caribbean plate motion investigations are examined.

  3. 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.

  4. 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.

  5. 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.

  6. 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…

  7. 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

  8. Finite Element Analysis of Magnetoelastic Plate Problems.

    DTIC Science & Technology

    1981-08-01

    deformation and in the incremental large deformation analysis, respectively. The classical Kirchhoff assumption of the undeformable normal to the midsurface is...current density , is constant across the thickness of the plate and is parallel to the midsurface of the plate; (2) the normal component of the

  9. Analytical Tripping Loads for Stiffened Plates

    DTIC Science & Technology

    1993-12-01

    extensional strains at the midsurface of the plate are negli- gible. 2 (vii) The incremental buckling displacements may be approximated by the...extensional strains at the midsurface of the plate are given by 3 el = ul + ! e12 = +VI + WIW2), e22 = V2 + 2 The strains in the beam are denoted by

  10. 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.

  11. 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.

  12. 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....

  13. 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....

  14. 30 CFR 22.10 - Approval plate.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 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. ___...

  15. 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…

  16. 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.

  17. 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. ___...

  18. Roll-forming tubes to header plates

    NASA Technical Reports Server (NTRS)

    Kramer, K.

    1976-01-01

    Technique has been developed for attaching and sealing tubes to header plates using a unique roll-forming tool. Technique is useful for attaching small tubes which are difficult to roll into conventional grooves in header plate tube holes, and for attaching when welding, brazing, or soldering is not desirable.

  19. 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.

  20. 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.

  1. Tectonics of oblique plate boundary systems

    NASA Astrophysics Data System (ADS)

    Díaz-Azpiroz, Manuel; Brune, Sascha; Leever, Karen A.; Fernández, Carlos; Czeck, Dyanna M.

    2016-12-01

    The relative displacement between lithospheric plates normally results in obliquely deforming plate boundaries. This is simply caused by the fact that, on plate tectonics basis, irregularly shaped plate boundaries are rarely perpendicular or parallel to small-circle rotation paths, which describe plate motion on a sphere (Fig. 1a). Global current relative plate motions estimated from geological data (DeMets et al., 2010; Argus et al., 2011) and GPS measurements (e.g., Kreemer et al., 2003; Argus et al., 2010) provide insight to the prevalent degrees of obliquity on Earth's surface. Based on these global data sets, Philippon and Corti (2016), statistically show that current orthogonal boundaries (obliquity angle smaller than 10°) represent around 8% of the total boundary length whereas strike-slip boundaries (obliquity angle larger than 80°) are encountered in < 10% of the total boundary length. Therefore, around 80% of active plate boundaries present oblique relative motions. Furthermore, changes in plate kinematics leading to migration or jumps in the rotation poles necessarily cause obliquity along former pure strike-slip or convergent/divergent boundaries (Fig. 1b).

  2. Evacuated flat-plate solar collectors

    SciTech Connect

    Whittemore, P.G.

    1981-09-15

    A structural support system is disclosed for use in an evacuated, flat-plate, solar collector to eliminate the problem of stress fractures in a glass cover plate. Nonlinearly spaced supports are used within the collector to dampen vibrations in the glass cover and to prevent overdeflection before buckling or stress fractures occur.

  3. Inverted flat plate solar collector. Final report

    SciTech Connect

    Brown, M.A.

    1981-08-26

    Construction and testing of an inverted flat plate solar collector are described. Heat transfer and economic analysis were performed to optimize the collector design. The newly designed collector was tested against two other flat plate collectors and the results and comparison of efficiencies are presented. (BCS)

  4. Fresnel diffraction plates are simple and inexpensive

    NASA Technical Reports Server (NTRS)

    Hoover, R. B.

    1967-01-01

    Fresnel plate demonstrates diffraction phenomena simply and inexpensively. A large number of identical diffracting apertures are made in random orientation on photographic film. When a small source of light is viewed through the plate, the diffraction pattern typical of the diffracting aperture is readily seen.

  5. Program sizes flange-tap orifice plate

    SciTech Connect

    Hogsett, J.E.

    1981-03-23

    A program developed for the TI-59 programmable calculator computes the differential pressure across an orifice, the gas flow through an orifice, or the orifice-plate bore for orifice plates with flange taps. Based on equations and data from L.K. Spinks, the program saves time in extracting from charts, tables, and graphs the values required to perform the calculations.

  6. 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.

  7. 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

  8. Bending equation for a quasianisotropic plate

    NASA Astrophysics Data System (ADS)

    Shachnev, V. A.

    2010-10-01

    In the framework of the linear theory of elasticity, an exact bending equation is obtained for the median plane of a plate whose material is a monoclinic system with the axis of symmetry perpendicular to the plate plane. As an example, the equation of the median plane of an isotropic plate is considered; the operator of this equation coincides with the operator of Sophie Germain's approximate equation. As the plate thickness tends to zero, the right-hand side of the equation is asymptotically equivalent to the right-hand side of the approximate equation. In addition, equations relating the median plane transverse stresses and the total stresses in the plate boundary planes to the median plane deflexions are obtained.

  9. Thyroid hormone and the growth plate.

    PubMed

    Shao, Yvonne Y; Wang, Lai; Ballock, R Tracy

    2006-12-01

    Thyroid hormone was first identified as a potent regulator of skeletal maturation at the growth plate more than forty years ago. Since that time, many in vitro and in vivo studies have confirmed that thyroid hormone regulates the critical transition between cell proliferation and terminal differentiation in the growth plate, specifically the maturation of growth plate chondrocytes into hypertrophic cells. However these studies have neither identified the molecular mechanisms involved in the regulation of skeletal maturation by thyroid hormone, nor demonstrated how the systemic actions of thyroid hormone interface with the local regulatory milieu of the growth plate. This article will review our current understanding of the role of thyroid hormone in regulating the process of endochondral ossification at the growth plate, as well as what is currently known about the molecular mechanisms involved in this regulation.

  10. 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.

  11. AE Source Orientation by Plate Wave Analysis

    NASA Technical Reports Server (NTRS)

    Gorman, Michael R.; Prosser, William H.

    1991-01-01

    Lead breaks (Hsu-Neilsen source) were used to generate simulated acoustic emission signals in an aluminum plate at angles of 0, 30, 60, and 90 degrees with respect to the plane of the plate. This was accomplished by breaking the lead on slots cut into the plate at the respective angles. The out-of-plane and in-plane displacement components of the resulting signals were detected by broad band transducers and digitized. Analysis of the waveforms showed them to consist of the extensional and flexural plate modes. The amplitude of both components of the two modes was dependent on the source orientation angle. This suggests that plate wave analysis may be used to determine the source orientation of acoustic emission sources.

  12. 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.

  13. 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.

  14. Drag of wings with end plates

    NASA Technical Reports Server (NTRS)

    Hemke, Paul E

    1928-01-01

    In this report a formula for calculating the induced drag of multiplanes with end plates is derived. The frictional drag of the end plates are used, is sufficiently large to increase the efficiency of the wing. Curves showing the reduction of drag for monoplanes and biplanes are constructed; the influence of gap-chord ratio, aspect ratio, and height of end plate are determined for typical cases. The method of obtaining the reduction of drag for a multiplane is described. Comparisons are made of calculated and experimental results obtained in wind tunnel tests with airfoils of various aspect ratios and end plates of various sizes. The agreement between calculated and experimental results is good. Analysis of the experimental results shows that the shape and section of the end plates are important.

  15. 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.

  16. Piezoactuation of sandwich plates with viscoelastic cores

    NASA Astrophysics Data System (ADS)

    Wang, Gang; Wereley, Norman M.

    1999-06-01

    Experimental and analytical validations of a Galerkin analysis of sandwich plates is presented in this paper. The 3-layered sandwich plate specimen consists of isotropic face-plates with surface bonded piezo-electric patch actuators, and a viscoelastic core. The experimental validation is conducted by testing sandwiched plates that are 67.31 cm (26.5') long, 52.07 cm (20.5') wide and nominally 0.16 cm (1/16') thick. The analysis includes the membrane and transverse energies in the face plates, and shear energies in the core. The shear modulus of the dissipative core is assumed to be complex and variant with frequency and temperature. The Golla-Hughes-McTavish (GHM) method is used to account for the frequency dependent properties of the viscoelastic core. Experiments have been conducted on sandwich plates with aluminum face-plates under clamped boundary conditions to validate the model for isotropic face-plates. Symmetric and asymmetric sandwiches have been tested. The maximum error in damped natural frequency predictions obtained via the assumed modes solutions is less than 11%. Analytical studies on the influence of the number of assumed modes in the Galerkin approximation, and the temperature variation, have been conducted. Error in the first plate bending mode is 112% when only a single in-plane mode is used; error reduces to 3.95% as the number of in-plane modes is increased to 25 in each of the in-plane directions. The study on the temperature influence shows that every plate mode has a corresponding temperature, wherein the loss factor is maximized.

  17. 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.

  18. The therapeutic effect of bone marrow-derived stem cell implantation after epiphyseal plate injury is abrogated by chondrogenic predifferentiation.

    PubMed

    Coleman, Rhima M; Schwartz, Zvi; Boyan, Barbara D; Guldberg, Robert E

    2013-02-01

    The goal of this study was to determine the effects of chondrogenic predifferentiation on the ability of bone marrow-derived stromal cells (BMSCs) delivered to growth plate defects to restore growth function. Chondrogenesis was induced with transforming growth factor (TGF)-β1 treatment in high-density monolayer cultures of BMSCs in vitro. The predifferentiated or undifferentiated BMSCs were either seeded into agarose gels for continued in vitro culture, or injected into growth plate defects via an in situ gelling agarose. Predifferentiated BMSCs had higher Sox-9, type II collagen, and aggrecan mRNA levels compared to undifferentiated cells after high-density monolayer culture. After transfer to agarose gels, predifferentiated cells did not produce a cartilaginous matrix, even with continued TGF-β1 stimulation, whereas undifferentiated cells produced a cartilaginous matrix in this system. Three-dimensional images of the growth plate created from microcomputed tomography scans showed that delivery of either predifferentiated or undifferentiated cells to defects resulted in a decrease in mineralized tether formation (fusion) in the growth plate tissue surrounding the defect to normal levels. Limb length discrepancy between injured and control limbs was corrected after treatment with undifferentiated, but not predifferentiated, cells. These results indicate that cell therapy may be an effective treatment to reduce growth dysfunction after growth plate injury, perhaps by maintaining the health of the uninjured growth plate tissue, and that the cell differentiation state plays a role in restoring the growth potential of the injured limb.

  19. Sonocatalytic injury of cancer cells attached on the surface of a nickel-titanium dioxide alloy plate.

    PubMed

    Ninomiya, Kazuaki; Maruyama, Hirotaka; Ogino, Chiaki; Takahashi, Kenji; Shimizu, Nobuaki

    2016-01-01

    The present study demonstrates ultrasound-induced cell injury using a nickel-titanium dioxide (Ni-TiO2) alloy plate as a sonocatalyst and a cell culture surface. Ultrasound irradiation of cell-free Ni-TiO2 alloy plates with 1 MHz ultrasound at 0.5 W/cm(2) for 30s led to an increased generation of hydroxyl (OH) radicals compared to nickel-titanium (Ni-Ti) control alloy plates with and without ultrasound irradiation. When human breast cancer cells (MCF-7 cells) cultured on the Ni-TiO2 alloy plates were irradiated with 1 MHz ultrasound at 0.5 W/cm(2) for 30s and then incubated for 48 h, cell density on the alloy plate was reduced to approximately 50% of the controls on the Ni-Ti alloy plates with and without ultrasound irradiation. These results indicate the injury of MCF-7 cells following sonocatalytic OH radical generation by Ni-TiO2. Further experiments demonstrated cell shrinkage and chromatin condensation after ultrasound irradiation of MCF-7 cells attached on the Ni-TiO2 alloy plates, indicating induction of apoptosis.

  20. 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.

  1. Nasopharyngeal culture

    MedlinePlus

    Culture - nasopharyngeal; Swab for respiratory viruses; Swab for staph carriage ... The test identifies viruses and bacteria that cause upper respiratory ... Staphylococcus aureus Methicillin-resistant Staphylococcus ...

  2. 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, ...

  3. Voltage-current characteristics of a pin-plate system with different plate configurations

    NASA Astrophysics Data System (ADS)

    Feng, Zhuangbo; Long, Zhengwei

    2013-03-01

    In this paper, the voltage-current (V-I) characteristics of a pin-plate system with four types of collection plate configurations are studied experimentally. The collection plates consider a single metal plate, a metal plate with a fly ash cake layer, a metal plate with a clean filter media and a metal plate with a dirty filter media. The results show that the clean filter media has no obvious effect on the V-I characteristics. But the dirty filter media reduces the current density because of its high resistance. The thick fly ash cake layer increase current density because of the anti-corona effect but the increment is not very obvious.

  4. 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.

  5. Comparative evaluation of phototrophic microtiter plate cultivation against laboratory-scale photobioreactors.

    PubMed

    Morschett, Holger; Schiprowski, Danny; Rohde, Jannis; Wiechert, Wolfgang; Oldiges, Marco

    2017-01-18

    Extended cultivation times, rendering phototrophic bioprocess development time inefficient, resulted in the recent development of micro-photobioreactors enabling accelerated process development. However, especially for laboratory photobioreactors, only little is known concerning the influence of design on process performance. Thus, the aim of the present investigation was to evaluate the scalability of a microtiter plate-based parallelized micro-photobioreactor against a representative set of established laboratory photobioreactors. Lipid production by Chlorella vulgaris was used as a model system. During exponential growth, the microtiter plate cultures achieved maximal growth rates of ca. 1.44 ± 0.02 day(-1) being in good agreement with the larger systems. Moreover, cultures in the micro-photobioreactor could be kept in the exponential phase up to the highest biomass concentrations most probably due to the beneficial light supply at this scale. Compared to the shake flask and test tube cultures, microtiter plate cultivation achieved an equivalent biomass yield, lipid content, and lipid fingerprint. In contrast, the flat-panel process resulted only in marginal productivity due to insufficient light supply. Thus, microtiter plates showed good scalability to the investigated laboratory photobioreactors as overall differences were rather small taking the differing scales into account.

  6. On the Origin of Plate Tectonics

    NASA Astrophysics Data System (ADS)

    Bercovici, D.

    2014-12-01

    The emergence of plate tectonics was Earth's defining moment. How and when platetectonics started is shrouded in mystery because of the paucity of observations in theArchean as well the challenge of understanding how plates are generated. The damage theoryof lithospheric weakening by grain-reduction provides a physical framework for plategeneration. This model builds on grain-scale processes to elucidate planetary-scaletectonics, and is consistent with lab and field observations of polycrystalline rocks andlithospheric shear zones. The grain-damage model accounts for the evolution of damage andhealing (by grain growth) at various planetary conditions, hence predicts plate boundaryformation and longevity, and how they depend on surface environment. For example, the onset of prototectonics is predicted to require clement conditions tokeep healing from erasing weak zones; conversely, cool conditions possibly requiredtectonics to draw down primordial CO2. Thus whether tectonics preceded a cool climate (andwater) or vice versa is immaterial as they likely needed each other or neither wouldexist. Sparse evidence that prototectonics co-initiated with liquid water hints at thelink between tectonics, water and surface conditions. The establishment of wide-spread plate tectonics started between >4Ga and 2.7Ga, and mayhave taken over a billion years to develop. Under Earth-like conditions, combininggrain-damage with intermittent Archean protosubduction produces persistent weak zones thataccumulate to yield well developed plates within 1Gyrs. In contrast, Venus' hottersurface conditions promotes healing and prohibits weak zone accumulation, which explainswhy plate tectonics failed to spread on our sister planet. Damage and weak-zone inheritance may also influence plate evolution and reorganization inthe modern era. Changes in plate direction, such as reflected in the Emperor-Hawaiianbend, leave weak zones misaligned with plate motion, causing oblique plate boundaries

  7. An updated digital model of plate boundaries

    NASA Astrophysics Data System (ADS)

    Bird, Peter

    2003-03-01

    A global set of present plate boundaries on the Earth is presented in digital form. Most come from sources in the literature. A few boundaries are newly interpreted from topography, volcanism, and/or seismicity, taking into account relative plate velocities from magnetic anomalies, moment tensor solutions, and/or geodesy. In addition to the 14 large plates whose motion was described by the NUVEL-1A poles (Africa, Antarctica, Arabia, Australia, Caribbean, Cocos, Eurasia, India, Juan de Fuca, Nazca, North America, Pacific, Philippine Sea, South America), model PB2002 includes 38 small plates (Okhotsk, Amur, Yangtze, Okinawa, Sunda, Burma, Molucca Sea, Banda Sea, Timor, Birds Head, Maoke, Caroline, Mariana, North Bismarck, Manus, South Bismarck, Solomon Sea, Woodlark, New Hebrides, Conway Reef, Balmoral Reef, Futuna, Niuafo'ou, Tonga, Kermadec, Rivera, Galapagos, Easter, Juan Fernandez, Panama, North Andes, Altiplano, Shetland, Scotia, Sandwich, Aegean Sea, Anatolia, Somalia), for a total of 52 plates. No attempt is made to divide the Alps-Persia-Tibet mountain belt, the Philippine Islands, the Peruvian Andes, the Sierras Pampeanas, or the California-Nevada zone of dextral transtension into plates; instead, they are designated as "orogens" in which this plate model is not expected to be accurate. The cumulative-number/area distribution for this model follows a power law for plates with areas between 0.002 and 1 steradian. Departure from this scaling at the small-plate end suggests that future work is very likely to define more very small plates within the orogens. The model is presented in four digital files: a set of plate boundary segments; a set of plate outlines; a set of outlines of the orogens; and a table of characteristics of each digitization step along plate boundaries, including estimated relative velocity vector and classification into one of 7 types (continental convergence zone, continental transform fault, continental rift, oceanic spreading ridge

  8. 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.

  9. 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

  10. 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.

  11. Thermal Casimir interaction between two magnetodielectric plates

    SciTech Connect

    Geyer, B.; Klimchitskaya, G. L.; Mostepanenko, V. M.

    2010-03-01

    We investigate the thermal Casimir interaction between two magnetodielectric plates made of real materials. On the basis of the Lifshitz theory, it is shown that for diamagnets and for paramagnets in the broad sense (with exception of ferromagnets) the magnetic properties do not influence the magnitude of the Casimir force. For ferromagnets, taking into account the realistic dependence of magnetic permeability on frequency, we conclude that the impact of magnetic properties on the Casimir interaction arises entirely from the contribution of the zero-frequency term in the Lifshitz formula. The computations of the Casimir free energy and pressure are performed for the configurations of two plates made of ferromagnetic metals (Co and Fe), for one plate made of ferromagnetic metal and the other of nonmagnetic metal (Au), for two ferromagnetic dielectric plates (on the basis of polystyrene), and for a ferromagnetic dielectric plate near a nonmagnetic metal plate. The dielectric permittivity of metals is described using both the Drude and the plasma model approaches. It is shown that the Casimir repulsion through the vacuum gap can be realized in the configuration of a ferromagnetic dielectric plate near a nonmagnetic metal plate described by the plasma model. In all cases considered, the respective analytical results in the asymptotic limit of large separations between the plates are obtained. The impact of the magnetic phase transition through the Curie temperature on the Casimir interaction is considered. In conclusion, we propose several experiments allowing to determine whether the magnetic properties really influence the Casimir interaction and to independently verify the Drude and plasma model approaches to the thermal Casimir force.

  12. 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;…

  13. Laminated anisotropic reinforced plastic plates and shells

    NASA Technical Reports Server (NTRS)

    Korolev, V. I.

    1981-01-01

    Basic technical theories and engineering calculation equations for anisotropic plates and shells made of rigid reinforced plastics, mainly laminated fiberglass, are presented and discussed. Solutions are given for many problems of design of structural plates and shells, including curved sections and tanks, as well as two chapters on selection of the optimum materials, are given. Accounting for interlayer shearing and transverse separation, which are new engineering properties, are discussed. Application of the results obtained to thin three ply plates and shells wth a light elastic filler is presented and discussed.

  14. Natural frequencies of twisted rotating plates

    NASA Astrophysics Data System (ADS)

    Ramamurti, V.; Kielb, R.

    1984-12-01

    A detailed comparison is presented of the predicted eigenfrequencies of twisted rotating plates as obtained by using two different shape functions. Primarily, rotating twisted plates of two different aspect ratios and two different thickness ratios are considered. The effects of rotation are included by using a "stress smoothing" technique when calculating the augmented stiffness matrix. In addition, the effects of Coriolis acceleration, contributions from membrane behaviour, setting angle and sweep angle are considered. The effects of geometric non-linearity are briefly discussed. Finally, results of a brief study of cambered plates are presented.

  15. Natural frequencies of twisted rotating plates

    NASA Technical Reports Server (NTRS)

    Ramamurti, V.; Kielb, R.

    1984-01-01

    A detailed comparison is presented of the predicted eigenfrequencies of twisted rotating plates as obtained by using two different shape functions. Primarily, rotating twisted plates of two different aspect ratios and two different thickness ratios are considered. The effects of rotation are included by using a 'stress smoothing' technique when calculating the augmented stiffness matrix. In addition, the effects of Coriolis acceleration, contributions from membrane behavior, setting angle and sweep angle are considered. The effects of geometric nonlinearity are briefly discussed. Finally, results of a brief study of cambered plates are presented.

  16. Research Investigation of Armor Plate Steels

    DTIC Science & Technology

    1942-02-20

    location at which the ballistic limits were determined. Ballistic limits and average hardness values from the 16 duplicate plates are given in Table 10 ...OF ARMOR PLATE STEELS Plate Nuzuber %C %.Mn %Si %Ni. %Cr %Mo %cu *7111 .46 1.58 .20 . 10 /.20 7276 .50 1.07 .3o 7277 .39 .35 .23 7278 .47 .91 .26 7279...26 2.3/2.7 7328 .34 .89 .20 1.3/1.7 .35/.46 * 7329 .34 .88 .25 1.3/1.7 10 /.20 7330 .46 .86 .24 .25/.35 : 10 /.20 734 .46 .84 .29 W 5. 35 7341 .34 .94

  17. Problem of Electromagnetoviscoelasticity for Multiply Connected Plates

    NASA Astrophysics Data System (ADS)

    Kaloerov, S. A.; Samodurov, A. A.

    2015-11-01

    A method for solving the problem of electromagnetoviscoelasticity for multiply connected plates is proposed. The small-parameter method is used to reduce this problem to a recursive sequence of problems of electromagnetoelasticity, which are solved by using complex potentials. A procedure is developed to determine, using complex potentials, approximations of the basic characteristics (stresses, electromagnetic-field strength, electromagnetic-flux density) of the electromagnetoelastic state at any time after application of a load. A plate with an elliptic hole is considered as an example. The variation in the electromagnetoelastic state of the multiply connected plate with time is studied

  18. Effusion plate using additive manufacturing methods

    DOEpatents

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

    2016-04-12

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

  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. Seismic anisotropy above a subducting plate

    SciTech Connect

    Shih, X.R.; Meyer, R.P. ); Schneider, J.F. )

    1991-08-01

    Shear-wave splitting observed in northeastern Colombia has provided evidence of seismic anisotropy in a shear zone immediately above a subducting plate. In an upper mantle composed mainly of olivine (57%) and orthopyroxene (17%), the splitting can be interpreted by wave propagation in an anisotropic medium of orthorhombic symmetry that results from alignment of these intrinsically anisotropic minerals. The mechanism of alignment is most likely the shearing associated with the subduction, aided by fluids migrating from the subducting plate when the plate exceeds 100 km in depth.