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Sample records for acetate membrane electrophoresis

  1. Fabricating PFPE Membranes for Capillary Electrophoresis

    NASA Technical Reports Server (NTRS)

    Lee, Michael C.; Willis, Peter A.; Greer, Frank; Rolland, Jason

    2009-01-01

    A process has been developed for fabricating perfluoropolyether (PFPE) membranes that contain microscopic holes of precise sizes at precise locations. The membranes are to be incorporated into laboratory-on-a-chip microfluidic devices to be used in performing capillary electrophoresis. The present process is a modified version of part of the process, described in the immediately preceding article, that includes a step in which a liquid PFPE layer is cured into solid (membrane) form by use of ultraviolet light. In the present process, one exploits the fact that by masking some locations to prevent exposure to ultraviolet light, one can prevent curing of the PFPE in those locations. The uncured PFPE can be washed away from those locations in the subsequent release and cleaning steps. Thus, holes are formed in the membrane in those locations. The most straightforward way to implement the modification is to use, during the ultraviolet-curing step, an ultraviolet photomask similar to the photomasks used in fabricating microelectronic devices. In lieu of such a photomask, one could use a mask made of any patternable ultraviolet-absorbing material (for example, an ink or a photoresist).

  2. Heterogeneity in human acid beta-glucosidase revealed by cellulose-acetate electrophoresis.

    PubMed

    Sa Miranda, M C; Aerts, J M; Pinto, R A; Magalhaes, J A; Barranger, J A; Tager, J M; Schram, A W

    1988-05-12

    Cellulose-acetate gel electrophoresis, a technique commonly used for the separation of human acid hydrolases, was applied to study heterogeneity in acid beta-glucosidase (EC 3.2.1.45). With this technique, three forms of beta-glucosidase were distinguishable in extracts of several tissues. The most anodic beta-glucosidase activity (band 3) represents the broad-specificity beta-glucosidase that is not deficient in Gaucher disease and is not inhibited by conduritol B-epoxide (CBE). The beta-glucosidase activity was deficient in Gaucher disease. A third beta-glucosidase activity with an intermediate mobility (band 2) was also inhibited by CBE and deficient in Gaucher disease. Band 1 and band 2 beta-glucosidase thus represent different forms of glucocerebrosidase. By adding phosphatidylserine and sphingolipid activator protein (SAP-2), monomeric glucocerebrosidase could be completely converted into a form that comigrated with band 2 beta-glucosidase of tissue extracts. The addition of phosphatidylserine only also resulted in a changed mobility of the monomeric enzyme, but the migration in this case differed from that of band 2 beta-glucosidase of tissue extracts. The electrophoretic profile of beta-glucosidase activity of tissue extracts changed upon ethanol/chloroform extraction: the two glucocerebrosidase forms were converted into a band with a mobility identical to that of band 1 beta-glucosidase. Our findings indicate that the interaction of glucocerebrosidase with phospholipid and SAP-2 has major effects on the mobility of the enzyme in the cellulose-acetate gel electrophoresis system. The findings with the cellulose-acetate gel electrophoretic system are discussed in relation to the heterogeneity in glucocerebrosidase observed with sucrose density gradient analysis, immunochemical methods and isoelectric focussing studies. PMID:3130106

  3. Continuous-flow electrophoresis: Membrane-associated deviations of buffer pH and conductivity

    NASA Technical Reports Server (NTRS)

    Smolka, A. J. K.; Mcguire, J. K.

    1978-01-01

    The deviations in buffer pH and conductivity which occur near the electrode membranes in continuous-flow electrophoresis were studied in the Beckman charged particle electrophoresis system and the Hanning FF-5 preparative electrophoresis instrument. The nature of the membranes separating the electrode compartments from the electrophoresis chamber, the electric field strength, and the flow rate of electrophoresis buffer were all found to influence the formation of the pH and conductivity gradients. Variations in electrode buffer flow rate and the time of electrophoresis were less important. The results obtained supported the hypothesis that a combination of Donnan membrane effects and the differing ionic mobilities in the electrophoresis buffer was responsible for the formation of the gradients. The significance of the results for the design and stable operation of continuous-flow electrophoresis apparatus was discussed.

  4. Dissolution control of Mg by cellulose acetate-polyelectrolyte membranes.

    PubMed

    Yliniemi, Kirsi; Wilson, Benjamin P; Singer, Ferdinand; Höhn, Sarah; Kontturi, Eero; Virtanen, Sannakaisa

    2014-12-24

    Cellulose acetate (CA)-based membranes are used for Mg dissolution control: the permeability of the membrane is adjusted by additions of the polyelectrolyte, poly(N,N-dimethylaminoethyl methacrylate) (PDMAEMA). Spin-coated films were characterized with FT-IR, and once exposed to an aqueous solution the film distends and starts acting as a membrane which controls the flow of ions and H2 gas. Electrochemical measurements (linear sweep voltammograms, open-circuit potential, and polarization) show that by altering the CA:PDMAEMA ratio the dissolution rate of Mg can be controlled. Such a control over Mg dissolution is crucial if Mg is to be considered as a viable, temporary biomedical implant material. Furthermore, the accumulation of corrosion products between the membrane and the sample diminishes the undesirable effects of high local pH and H2 formation which takes place during the corrosion process. PMID:25426707

  5. Ion selective permeation through cellulose acetate membranes in forward osmosis.

    PubMed

    Irvine, Gavin J; Rajesh, Sahadevan; Georgiadis, Michael; Phillip, William A

    2013-12-01

    Solute-solute interactions can have a dramatic impact on the permeation of solutes through dense polymeric membranes. In particular, understanding how solute-solute interactions can affect the design of osmotically driven membrane processes (ODMPs) is critical to the successful development of these emerging water treatment and energy generation processes. In this work, we investigate the influence that solute-solute interactions have on nitrate permeation through an asymmetric cellulose acetate forward osmosis membrane. A series of experiments that included systematic modifications to the cation paired with nitrate, the identity of the draw solute, and the solution pH were conducted. These experiments reveal that in the unique operating geometry of ODMPs, where solute containing solutions are present on both sides of the membrane, nitrate fluxes are significantly higher (>15 times in some cases) than predicted by existing models for solute permeation in ODMPs. The identity of the cation paired with nitrate influences the flux of nitrate; the identity of the cation in the draw solution does not affect the flux of nitrate; however, the identity of the anion in the draw solution has the most significant impact on the flux of nitrate. These results suggest that an ion exchange mechanism, which allows nitrate to switch rapidly with anions from the draw solution, is present when cellulose acetate based membranes are used in ODMPs. PMID:24152190

  6. Nanoporous layered silicate AMH-3/cellulose acetate nanocomposite membranes for gas separations

    E-print Network

    Nair, Sankar

    Nanoporous layered silicate AMH-3/cellulose acetate nanocomposite membranes for gas separations Wun. The exfoliated SAMH-3 flakes were used to form SAMH-3/cellulose acetate (CA) membranes. Their micro- structure of each type of material [4]. Various inorganic porous materials such as zeolite, metal oxide nanotubes

  7. Membrane interfaces for sample introduction in capillary zone electrophoresis

    SciTech Connect

    Bao, L.; Dasgupta, P.K.

    1992-05-01

    Small lengths of narrow-bore tubular membranes can be interposed in the separation capillary in capillary electrophoretic separation systems. These membrane segments can be used as sampling interfaces; a jacket is built outside the membrane, and the sample is introduced by diffusion/permeation through the membrane. Various examples are shown; the determination of gaseous samples through a porous membrane, the determination of ionizable/nonionic solutes by permeation through a silicone rubber membrane, and the separation of low MW constituents in blood plasma by transport through a dialysis membrane. In the first two cases, significant preconcentration is possible, thus permitting attractive detection limits. 23 refs., 10 figs., 2 tabs.

  8. Tunable Membranes for Free-Flow Zone Electrophoresis in PDMS Microchip Using Guided Self-Assembly of Silica Microbeads

    E-print Network

    Song, Yong-Ak

    In this paper, we evaluate the strategy of using self-assembled microbeads to build a robust and tunable membrane for free-flow zone electrophoresis in a PDMS microfluidic chip. To fabricate a porous membrane as a salt ...

  9. Continuous Ethanol Production with a Membrane Bioreactor at High Acetic Acid Concentrations

    PubMed Central

    Ylitervo, Päivi; Franzén, Carl Johan; Taherzadeh, Mohammad J.

    2014-01-01

    The release of inhibitory concentrations of acetic acid from lignocellulosic raw materials during hydrolysis is one of the main concerns for 2nd generation ethanol production. The undissociated form of acetic acid can enter the cell by diffusion through the plasma membrane and trigger several toxic effects, such as uncoupling and lowered intracellular pH. The effect of acetic acid on the ethanol production was investigated in continuous cultivations by adding medium containing 2.5 to 20.0 g·L?1 acetic acid at pH 5.0, at a dilution rate of 0.5 h?1. The cultivations were performed at both high (~25 g·L?1) and very high (100–200 g·L?1) yeast concentration by retaining the yeast cells inside the reactor by a cross-flow membrane in a membrane bioreactor. The yeast was able to steadily produce ethanol from 25 g·L?1 sucrose, at volumetric rates of 5–6 g·L?1·h?1 at acetic acid concentrations up to 15.0 g·L?1. However, the yeast continued to produce ethanol also at a concentration of 20 g·L?1 acetic acid but at a declining rate. The study thereby demonstrates the great potential of the membrane bioreactor for improving the robustness of the ethanol production based on lignocellulosic raw materials. PMID:25028956

  10. Investigation of the pore structure and morphology of cellulose acetate membranes using small-angle neutron scattering. 1: Cellulose acetate active layer membranes

    SciTech Connect

    Kulkarni, S.; Krause, S. ); Wignall, G.D. . Solid State Div.); Hammouda, B. . Center for High Resolution Neutron Scattering)

    1994-11-07

    The structure of ultrathin cellulose acetate membranes, known as active layer membranes, has been investigated using small-angle neutron scattering. These membranes are known to have structural and functional similarity to the surface or skin layer in commercial reverse-osmosis (RO) membranes and hence are useful model systems for understanding the structure of the RO membrane skin layer. Active layer membranes were studied after swelling them with either D[sub 2]O or CD[sub 3]OD. The results in both cases clearly indicated the presence of very small (10--20 [angstrom]) porous structures in the membrane. The presence of such pores has been a subject of long-standing controversy in this area. The data were analyzed using a modified Debye-Bueche analysis and the resultant membrane structure was seen to agree well with structural information from electron microscopic studies. Finally, a possible explanation for the differences in scattering observed between the D[sub 2]O swollen membranes and the CD[sub 3]OD swollen membranes has been presented.

  11. Performance of cellulose acetate butyrate membranes in hyperfiltration of sodium chloride and urea feed solution

    NASA Technical Reports Server (NTRS)

    Wydeven, T.; Leban, M.

    1973-01-01

    Cellulose acetate butyrate (CAB) membranes are shown to give high salt and urea rejection with water flux of about 3 gallons/sq ft per day at 600 psig. Membranes prepared from a formulation containing glyoxal show a significant increase in flux and decrease in salt and urea rejection with drying time. Zero drying time gives maximum urea and salt rejection and is therefore most suitable for hyperfiltration of sodium chloride and urea feed solution.

  12. Non-denaturing gel electrophoresis system for the purification of membrane bound proteins

    SciTech Connect

    Cavinato, A.G.; Macleod, R.M.; Ahmed, M.S.

    1988-01-01

    A new method is described for the purification of a membrane bound glycoprotein, the kappa opioid receptor from human placental tissue. The method uses preparative slab-gel electrophoresis in the presence of the non-denaturing detergent CHAPS. A linear relationship between log molecular weight and SDS PAGE electrophoretic mobility of known molecular weight markers, in the presence of CHAPS, is observed. Using this method, we were able partially to purify an /sup 3/H-etorphine binding glycoprotein, from placental villus tissue, with an apparent molecular weight range of 60-70,000. The iodinated glycoprotein migrates in SDS PAGE with an apparent molecular weight of 63,000. This method may be useful for the isolation of membrane bound proteins, especially when an affinity ligand is not available.

  13. Alginate fouling reduction of functionalized carbon nanotube blended cellulose acetate membrane in forward osmosis.

    PubMed

    Choi, Hyeon-Gyu; Son, Moon; Yoon, SangHyeon; Celik, Evrim; Kang, Seoktae; Park, Hosik; Park, Chul Hwi; Choi, Heechul

    2015-10-01

    Functionalized multi-walled carbon nanotube blended cellulose acetate (fCNT-CA) membranes were synthesized for forward osmosis (FO) through phase inversion. The membranes were characterized through SEM, FTIR, and water contact angle measurement. AFM was utilized to investigate alginate fouling mechanism on the membrane. It reveals that the fCNT contributes to advance alginate fouling resistance in FO (57% less normalized water flux decline for 1% fCNT-CA membrane was observed than that for bare CA membrane), due to enhanced electrostatic repulsion between the membrane and the alginate foulant. Furthermore, it was found that the fCNT-CA membranes became more hydrophilic due to carboxylic groups in functionalized carbon nanotube, resulting in approximately 50% higher water-permeated flux than bare CA membrane. This study presents not only the fabrication of fCNT-CA membrane and its application to FO, but also the quantification of the beneficial role of fCNT with respect to alginate fouling in FO. PMID:26022283

  14. Removal of acetic acid from simulated hemicellulosic hydrolysates by emulsion liquid membrane with organophosphorus extractants.

    PubMed

    Lee, Sang Cheol

    2015-09-01

    Selective removal of acetic acid from simulated hemicellulosic hydrolysates containing xylose and sulfuric acid was attempted in a batch emulsion liquid membrane (ELM) system with organophosphorus extractants. Various experimental variables were used to develop a more energy-efficient ELM process. Total operation time of an ELM run with a very small quantity of trioctylphosphine oxide as the extractant was reduced to about a third of those required to attain almost the same extraction efficiency as obtained in previous ELM works without any extractant. Under specific conditions, acetic acid was selectively separated with a high degree of extraction and insignificant loss of xylose, and its purity and enrichment ratio in the stripping phase were higher than 92% and 6, respectively. Also, reused organic membrane solutions exhibited the extraction efficiency as high as fresh organic solutions did. These results showed that the current ELM process would be quite practical. PMID:26056774

  15. Synthesis of polymer electrolyte membranes from cellulose acetate/poly(ethylene oxide)/LiClO4 for lithium ion battery application

    NASA Astrophysics Data System (ADS)

    Nurhadini, Arcana, I. Made

    2015-09-01

    This study was conducted to determine the effect of cellulose acetate on poly(ethylene oxide)-LiClO4 membranes as the polymer electrolyte. Cellulose acetate is used as an additive to increase ionic conductivity and mechanical property of polymer electrolyte membranes. The increase the percentage of cellulose acetate in membranes do not directly effect on the ionic conductivity, and the highest ionic conductivity of membranes about 5,7 × 10-4 S/cm was observed in SA/PEO/LiClO4 membrane with cellulose ratio of 10-25% (w/w). Cellulose acetate in membranes increases mechanical strength of polymer electrolyte membranes. Based on TGA analysis, this polymer electrolyte thermally is stable until 270 °C. The polymer electrolyte membrane prepared by blending the cellulose acetate, poly(ethylene oxide), and lithium chlorate could be potentially used as a polymer electrolyte for lithium ion battery application.

  16. Post radiation grafting of vinyl acetate onto low density polyethylene films: Preparation and properties of membrane

    NASA Astrophysics Data System (ADS)

    M. Dessouki, Ahmed

    Reverse osmosis membranes were prepared by the post radiation grafting of vinyl acetate onto low density polyethylene films. The factors affecting the grafting process such as radiation dose, monomer concentration and temperature on the grafting yield were studied. It was found that the dependence of the grafting rate on radiation intensity and monomer concentration was found to be of 0.64 and 1.4 order, respectively. The activation energy for this grafting system was calculated and found to be 4.45 kcal/mol above 30°C. Some properties of the grafted films such as specific electric resistance, water uptake, mechanical properties and thermal and chemical stability were investigated. An improvement in these properties was observed which makes possible the use of these membranes in some practical applications. The use of such membranes for reverse osmosis desalination of saline water was tested. The effect of operating time, degree of grafting and applied pressure on the water flux and salt rejection were determined. The results showed salt rejection percent over 90% and a reasonable water flux. A suitable degree of grafting of the membrane was determined as well as the optimum applied pressure.

  17. Investigation of the pore structure and morphology of cellulose acetate membranes using small-angle neutron scattering. 2: Ultrafiltration and reverse-osmosis membranes

    SciTech Connect

    Kulkarni, S.; Krause, S. ); Wignall, G.D. . Solid State Div.)

    1994-11-07

    Pore structure in cellulose acetate ultrafiltration (UF) and reverse-osmosis (RO) membranes has been studied using small-angle neutron scattering. Scattering experiments were carried out on dry membranes as well as on membranes swollen with deuterated solvents (D[sub 2]O and CD[sub 3]OD). In addition, the RO membranes were studied both before and after annealing (a process of heating a membrane in a water bath at [approximately]75 C to improve its separation properties). The pore surface in UF membranes was found to be smooth and nonfractal, as evidenced by the fourth power law behavior at high Q. Values of average pore sizes obtained for dry and solvent swollen membranes agree well with pore sizes obtained by other methods. For cellulose acetate RO membranes in their dry state, the unannealed membrane appears to consist of two discrete pore size distributions in the intermediate and high Q region while the annealed membrane contains a much wider distribution of pore sizes. These results give a good account of the changes occurring in the structure of RO membranes as a result of annealing, and agree well with the prediction of other authors.

  18. Sensitivity enhancement in direct coupling of supported liquid membrane extractions to capillary electrophoresis by means of transient isotachophoresis and large electrokinetic injections.

    PubMed

    Pant??ková, Pavla; Kubá?, Pavel; Bo?ek, Petr

    2015-04-10

    Enhanced sensitivity for determination of basic drugs in body fluids was achieved by in-line coupling of extraction across supported liquid membrane (SLM) to large electrokinetic injection and transient isotachophoresis-capillary zone electrophoresis (tITP-CZE) in commercial CZE instrument. Twelve cm long tITP plug of 300mM ammonium acetate was formed in the separation capillary just before the electrokinetic injection of acceptor solution containing nortriptyline, haloperidol and loperamide extracted across the SLM. The tITP plug ensured efficient stacking and preconcentration of the injected basic drugs due to the tITP action of ammonium and the drugs were then separated by CZE using 5.2M acetic acid as background electrolyte. No interferences were observed from highly-abundant body fluid species (NaCl and human serum albumin) due to the excellent clean-up properties of SLMs and analytical sensitivity increased up to 340 times compared to SLM extractions coupled in-line to CZE with standard hydrodynamic injections. The SLM-tITP-CZE method was characterized by good repeatability (RSDs of peak areas below 7.8%), linearity over two orders of magnitude (r(2) better than 0.994) and limits of detection (defined as 3×S/N) between 3 and 45?g/L. Interfacing of SLM extractions to CZE instrumentation was achieved by low-cost, disposable micro-extraction devices, which can be routinely prepared in every analytical laboratory. These devices eliminated sample carry-over, minimized the need for manual sample handling and ensured fully automated determination (including extraction, injection, preconcentration and separation) of the three basic drugs in 20?L of untreated body fluids. PMID:25747667

  19. Net Increase of platelet membrane tyrosine specific-protein kinase activity by phorbol myristate acetate

    SciTech Connect

    Ishihara, Noriko; Sakamoto, Hikaru; Iwama, Minako; Kobayashi, Bonro )

    1990-01-01

    Tyrosine protein kinase (TPK) activity in rabbit platelets after stimulation by phorbol myristate acetate (PMA) or thrombin was directly estimated by {sup 32}P incorporation from ({gamma}-{sup 32})ATP into synthetic peptide angiotensin II. By PMA-treatment a net increase of TPK activity was obtained, while thrombin acted on the TPK quickly but stimulation was limited within the range attained by the control after lengthy incubation. The responsive TPK to these stimulators was localized mainly in membrane but much less in cytosol. The specific activity of the particulate TPK was low in the sonicate of control ice cold platelets but increased about 6-fold when the platelets were incubated at 37{degree}C. On a brief contact of platelets with PMA at 37{degrees}C the TPK was fully activated and reached a maximum value about 130% of the control. Determination of phosphotyrosine phosphatase in the stimulated platelet sonicate revealed that its participation in the above described increase of {sup 32}P-incorporation was meagre. The quick response suggested a possible role of TPK in the signal transduction through the platelet cell membrane.

  20. Fouling propensity and separation efficiency of epoxidated polyethersulfone incorporated cellulose acetate ultrafiltration membrane in the retention of proteins

    NASA Astrophysics Data System (ADS)

    Jayalakshmi, A.; Rajesh, S.; Mohan, D.

    2012-10-01

    Epoxidated polyethersulfone (EPES) incorporated cellulose acetate (CA) ultrafiltration membranes were prepared by diffusion induced precipitation technique in the absence and presence of pore former polyethyleneglycol-600. Effect of blend ratio on the compatibility, thermal stability, mechanical strength, hydrophilicity, morphology, pure water flux, protein adsorption resistance, protein separation efficiency and fouling propensity of the CA/EPES blend membranes was evaluated. Addition of EPES results in the formation of thin separating layer and spongy sub layer in CA/EPES blend membranes. The efficiency of these membranes in the separation of commercially important proteins such as bovine serum albumin, egg albumin, pepsin and trypsin was studied and found to be enhanced as compared to CA membranes. The fouling-resistant capability of the membranes was studied by bovine serum albumin as the model foulant and flux recovery ratio of the membranes were calculated. Attempts have been made to correlate the changes in membrane morphology with pure water flux, hydraulic resistance, thermal and mechanical stability, separation efficiency and antifouling property of the CA/EPES membranes. The optimal combination of CA and EPES, thus allows the preparation of high performance UF membranes which are sufficiently dense to retain proteins and at the same time give economically viable fluxes.

  1. Magnetic Targeted Delivery of Dexamethasone Acetate across the Round Window Membrane in Guinea Pigs

    PubMed Central

    Du, Xiaoping; Chen, Kejian; Kuriyavar, Satish; Kopke, Richard D.; Grady, Brian P.; Bourne, David H.; Li, Wei; Dormer, Kenneth J.

    2012-01-01

    Hypothesis Magnetically susceptible PLGA nanoparticles will effectively target the round window membrane (RWM) for delivery of dexamethasone-acetate (Dex-Ac) to the scala tympani. Background Targeted delivery of therapeutics to specific tissues can be accomplished using different targeting mechanisms. One technology includes iron oxide nanoparticles, susceptible to external magnetic fields. If a nanocomposite composed of biocompatible polymer (PLGA), magnetite, and Dex-Ac can be pulled into and across the mammalian RWM, drug delivery can be enhanced. Method In vitro targeting and release kinetics of PLGA-magnetite-Dex-Ac nanoparticles first were measured using a RWM model. Next, these optimized nanocomposites were targeted to the RWM by filling the niche in anesthetized guinea pigs. A permanent magnet was placed opposite the RWM for 1 hour. Cochlear soft tissues, perilymph, and RWM were harvested after euthanasia and steroid levels were measured using HPLC. Results Membrane transport, in vitro, proved optimal targeting using a lower particle magnetite concentration (1 versus 5 or 10 mg/ml). In vivo targeted PLGA-magnetite-Dex-Ac particles had an average size of 482.8 ± 158 nm (DLS) and an average zeta potential ?19.9 ± 3.3 mV. In 1 hour, there was significantly increased cochlear targeted delivery of Dex or Dex-Ac, compared with diffusion alone. Conclusion Superparamagnetic PLGA-magnetite-Dex-Ac nanoparticles under an external magnetic field (0.26 mT) for 1 hour significantly increased Dex-Ac delivery to the inner ear. The RWM was not completely permeated and also became loaded with nanocomposites, indicating that delivery to the cochlea would continue for weeks by PLGA degradation and passive diffusion. PMID:23187928

  2. Two-dimensional electrophoresis of plasma membranes, showing differences among wild-type and abnormal ascospore mutant strains of Neurospora crassa.

    PubMed Central

    Nasrallah, J B; Srb, A M

    1983-01-01

    Plasma membranes isolated from vegetative cultures of wild-type Neurospora crassa were analyzed by two-dimensional electrophoresis, followed by staining with silver nitrate to visualize proteins and fluorescein-labeled concanavalin A to visualize glycosylated subunits. Mycelial plasma membranes from strains carrying mutations affecting ascospores were also analyzed. Two of the mutant strains were shown to have aberrant two-dimensional membrane subunit patterns. The correlation of these abnormalities with the known electron microscopic evidence for aberrations of their ascospore-delimiting membrane during ascospore genesis is discussed. Images PMID:6224773

  3. Fabrication of tethered carbon nanotubes in cellulose acetate/polyethylene glycol-400 composite membranes for reverse osmosis.

    PubMed

    Sabir, Aneela; Shafiq, Muhammad; Islam, Atif; Sarwar, Afsheen; Dilshad, Muhammad Rizwan; Shafeeq, Amir; Zahid Butt, Muhammad Taqi; Jamil, Tahir

    2015-11-01

    In this study pristine multi-walled carbon nanotubes (MWCNTs) were surface engineered (SE) in strong acidic medium by oxidation purification method to form SE-MWCNT. Five different amount of SE-MWCNT ranging from 0.1 to 0.5 wt% were thoroughly and uniformly dispersed in cellulose acetate/polyethylene glycol (CA/PEG400) polymer matrix during synthesis of membrane by dissolution casting method. The structural analysis, surface morphology and roughness was carried out by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and atomic force microscopy (AFM), respectively, which showed that the dispersed SE-MWCNT was substantially tethered in CA/PEG400 polymer matrix membrane. The thermogravimetric analysis (TGA) of membranes also suggested some improvement in thermal properties with the addition of SE-MWCNT. Finally, the performance of these membranes was assessed for suitability in drinking water treatment. The permeation flux and salt rejection were determined by using indigenously fabricated reverse osmosis pilot plant with 1000 ppm NaCl feed solution. The results showed that the tethered SE-MWCNT/CA/PEG400 polymer matrix membrane, with strong SE-MWCNTs/polymer matrix interaction, improved the salt rejection performance of the membrane with the salt rejection of 99.8% for the highest content of SE-MWCNT. PMID:26256386

  4. Diffusive transfer to membranes as an effective interface between gel electrophoresis and mass spectrometry

    NASA Astrophysics Data System (ADS)

    Ogorzalek Loo, Rachel R.; Mitchell, Charles; Stevenson, Tracy I.; Loo, Joseph A.; Andrews, Philip C.

    1997-12-01

    Diffusive transfer was examined as a blotting method to transfer proteins from polyacrylamide gels to membranes for ultraviolet matrix-assisted laser desorption ionization (MALDI) mass spectrometry. The method is well-suited for transfers from isoelectric focusing (IEF) gels. Spectra have been obtained for 11 pmol of 66 kDa albumin loaded onto an IEF gel and subsequently blotted to polyethylene. Similarly, masses of intact carbonic anhydrase and hemoglobin were obtained from 14 and 20 pmol loadings. This methodology is also compatible with blotting high molecular weight proteins, as seen for 6 pmol of the 150 kDa monoclonal antibody anti-[beta]-galactosidase transferred to Goretex. Polypropylene, Teflon, Nafion and polyvinylidene difluoride (PVDF) also produced good spectra following diffusive transfer. Only analysis from PVDF required that the membrane be kept wet prior to application of matrix. Considerations in mass accuracy for analysis from large-area membranes with continuous extraction and delayed extraction were explored, as were remedies for surface charging. Vapor phase CNBr cleavage was applied to membrane-bound samples for peptide mapping.

  5. Electrophoresis and isoelectric focusing of whole cell and membrane proteins from the extremely halophilic archaebacteria

    NASA Technical Reports Server (NTRS)

    Stan-Lotter, Helga; Lang, Frank J., Jr.; Hochstein, Lawrence I.

    1989-01-01

    The subunits from two purified halobacterial membrane enzymes (ATPase and nitrate reductase) behaved differently with respect to isoelectric focusing, silver staining and interaction with ampholytes. Differential behavior was also observed in whole cell proteins from Halobacterium saccharovorum regarding resolution in two-dimensional gels and silver staining. It is proposed that these differences reflect the existence of two classes of halobacterial proteins.

  6. ADSORPTION AND MEMBRANE SEPARATION MEASUREMENTS WITH MIXTURES OF ETHANOL, ACETIC ACID, AND WATER

    EPA Science Inventory

    Biomass fermentation produces ethanol and other renewable biofuels. Pervaporation using hydrophobic membranes is potentially a cost-effective means of removing biofuels from fermentation broths for small- to medium-scale applications. Silicalite-filled polydimethylsiloxane (PDMS)...

  7. Conjugation of silica nanoparticles with cellulose acetate/polyethylene glycol 300 membrane for reverse osmosis using MgSO4 solution.

    PubMed

    Sabir, Aneela; Shafiq, Muhammad; Islam, Atif; Jabeen, Faiza; Shafeeq, Amir; Ahmad, Adnan; Zahid Butt, Muhammad Taqi; Jacob, Karl I; Jamil, Tahir

    2016-01-20

    Thermally-induced phase separation (TIPS) method was used to synthesize polymer matrix (PM) membranes for reverse osmosis from cellulose acetate/polyethylene glycol (CA/PEG300) conjugated with silica nanoparticles (SNPs). Experimental data showed that the conjugation of SNPs changed the surface properties as dense and asymmetric composite structure. The results were explicitly determined by the permeability flux and salt rejection efficiency of the PM-SNPs membranes. The effect of SNPs conjugation on MgSO4 salt rejection was more significant in magnitude than on permeation flux i.e. 2.38L/m(2)h. FTIR verified that SNPs were successfully conjugated on the surface of PM membrane. DSC of PM-SNPs shows an improved Tg from 76.2 to 101.8°C for PM and PM-S4 respectively. Thermal stability of the PM-SNPs membranes was observed by TGA which was significantly enhanced with the conjugation of SNPs. The micrographs of SEM and AFM showed the morphological changes and increase in the valley and ridges on membrane surface. Experimental data showed that the PM-S4 (0.4wt% SNPs) membrane has maximum salt rejection capacity and was selected as an optimal membrane. PMID:26572387

  8. Versatile microanalytical system with porous polypropylene capillary membrane for calibration gas generation and trace gaseous pollutants sampling applied to the analysis of formaldehyde, formic acid, acetic acid and ammonia in outdoor air.

    PubMed

    Coelho, Lúcia H G; Melchert, Wanessa R; Rocha, Flavio R; Rocha, Fábio R P; Gutz, Ivano G R

    2010-11-15

    The analytical determination of atmospheric pollutants still presents challenges due to the low-level concentrations (frequently in the ?g m(-3) range) and their variations with sampling site and time. In this work, a capillary membrane diffusion scrubber (CMDS) was scaled down to match with capillary electrophoresis (CE), a quick separation technique that requires nothing more than some nanoliters of sample and, when combined with capacitively coupled contactless conductometric detection (C(4)D), is particularly favorable for ionic species that do not absorb in the UV-vis region, like the target analytes formaldehyde, formic acid, acetic acid and ammonium. The CMDS was coaxially assembled inside a PTFE tube and fed with acceptor phase (deionized water for species with a high Henry's constant such as formaldehyde and carboxylic acids, or acidic solution for ammonia sampling with equilibrium displacement to the non-volatile ammonium ion) at a low flow rate (8.3 nL s(-1)), while the sample was aspirated through the annular gap of the concentric tubes at 2.5 mL s(-1). A second unit, in all similar to the CMDS, was operated as a capillary membrane diffusion emitter (CMDE), generating a gas flow with know concentrations of ammonia for the evaluation of the CMDS. The fluids of the system were driven with inexpensive aquarium air pumps, and the collected samples were stored in vials cooled by a Peltier element. Complete protocols were developed for the analysis, in air, of NH(3), CH(3)COOH, HCOOH and, with a derivatization setup, CH(2)O, by associating the CMDS collection with the determination by CE-C(4)D. The ammonia concentrations obtained by electrophoresis were checked against the reference spectrophotometric method based on Berthelot's reaction. Sensitivity enhancements of this reference method were achieved by using a modified Berthelot reaction, solenoid micro-pumps for liquid propulsion and a long optical path cell based on a liquid core waveguide (LCW). All techniques and methods of this work are in line with the green analytical chemistry trends. PMID:21035648

  9. Gypsum (CaSO4·2H2O) Scaling on Polybenzimidazole and Cellulose Acetate Hollow Fiber Membranes under Forward Osmosis.

    PubMed

    Chen, Si Cong; Su, Jincai; Fu, Feng-Jiang; Mi, Baoxia; Chung, Tai-Shung

    2013-01-01

    We have examined the gypsum (CaSO4·2H2O) scaling phenomena on membranes with different physicochemical properties in forward osmosis (FO) processes. Three hollow fiber membranes made of (1) cellulose acetate (CA), (2) polybenzimidazole (PBI)/polyethersulfone (PES) and (3) PBI-polyhedral oligomeric silsesquioxane (POSS)/polyacrylonitrile (PAN) were studied. For the first time in FO processes, we have found that surface ionic interactions dominate gypsum scaling on the membrane surface. A 70% flux reduction was observed on negatively charged CA and PBI membrane surfaces, due to strong attractive forces. The PBI membrane surface also showed a slightly positive charge at a low pH value of 3 and exhibited a 30% flux reduction. The atomic force microscopy (AFM) force measurements confirmed a strong repulsive force between gypsum and PBI at a pH value of 3. The newly developed PBI-POSS/PAN membrane had ridge morphology and a contact angle of 51.42° ± 14.85° after the addition of hydrophilic POSS nanoparticles and 3 min thermal treatment at 95 °C. Minimal scaling and an only 1.3% flux reduction were observed at a pH value of 3. Such a ridge structure may reduce scaling by not providing a locally flat surface to the crystallite at a pH value of 3; thus, gypsum would be easily washed away from the surface. PMID:24957062

  10. STABILITY OF MFI ZEOLITE-FILLED PDMS MEMBRANES DURING PERVAPORATIVE ETHANOL RECOVERY FROM AQUEOUS MIXTURES CONTAINING ACETIC ACID

    EPA Science Inventory

    Pervaporation is a potential process for recovering bioethanol produced from biomass fermentation. Fermentation broths contain ethanol, water, and a variety of other compounds, often including carboxylic acids. The effects of acetic acid on long-term pervaporation of aqueous et...

  11. Analysis of electrophoresis performance

    NASA Technical Reports Server (NTRS)

    Roberts, G. O.

    1984-01-01

    The SAMPLE computer code models electrophoresis separation in a wide range of conditions. Results are included for steady three dimensional continuous flow electrophoresis (CFE), time dependent gel and acetate film experiments in one or two dimensions and isoelectric focusing in one dimension. The code evolves N two dimensional radical concentration distributions in time, or distance down a CFE chamber. For each time or distance increment, there are six stages, successively obtaining the pH distribution, the corresponding degrees of ionization for each radical, the conductivity, the electric field and current distribution, and the flux components in each direction for each separate radical. The final stage is to update the radical concentrations. The model formulation for ion motion in an electric field ignores activity effects, and is valid only for low concentrations; for larger concentrations the conductivity is, therefore, also invalid.

  12. Proteins Induced during Adaptation of Acetobacter aceti to High Acetate Concentrations

    PubMed Central

    Steiner, Peter; Sauer, Uwe

    2001-01-01

    As a typical product of microbial metabolism, the weak acid acetate is well known for its cytotoxic effects. In contrast to most other microbes, the so-called acetic acid bacteria can acquire significant resistance to high acetate concentrations when properly adapted to such hostile conditions. To characterize the molecular events that are associated with this adaptation, we analyzed global protein expression levels during adaptation of Acetobacter aceti by two-dimensional gel electrophoresis. Adaptation was achieved by using serial batch and continuous cultivations with increasing acetate supplementation. Computer-aided analysis revealed a complex proteome response with at least 50 proteins that are specifically induced by adaptation to acetate but not by other stress conditions, such as heat or oxidative or osmotic stress. Of these proteins, 19 were significantly induced in serial batch and continuous cultures and were thus noted as acetate adaptation proteins (Aaps). Here we present first microsequence information on such Aaps from A. aceti. Membrane-associated processes appear to be of major importance for adaptation, because some of the Aap bear N-terminal sequence homology to membrane proteins and 11 of about 40 resolved proteins from membrane protein-enriched fractions are significantly induced. PMID:11722895

  13. Synthesis and Characterisation of ETS-10/Acetate-based Ionic Liquid/Chitosan Mixed Matrix Membranes for CO2/N2 Permeation

    PubMed Central

    Casado-Coterillo, Clara; López-Guerrero, María del Mar; Irabien, Ángel

    2014-01-01

    Mixed matrix membranes (MMMs) were prepared by incorporating organic surfactant-free hydrothermally synthesised ETS-10 and 1-ethyl-3-methylimidazolium acetate ionic liquid (IL) to chitosan (CS) polymer matrix. The membrane material characteristics and permselectivity performance of the two-component membranes were compared with the three-component membrane and the pure CS membrane. The addition of IL increased CO2 solubility of the polymer, and, thus, the CO2 affinity was maintained for the MMMs, which can be correlated with the crystallinity, measured by FT-IR, and void fraction calculations from differences between theoretical and experimental densities. The mechanical resistance was enhanced by the ETS-10 nanoparticles, and flexibility decreased in the two-component ETS-10/CS MMMs, but the flexibility imparted by the IL remained in three-component ETS-10/IL/CS MMMs. The results of this work provide insight into another way of facing the adhesion challenge in MMMs and obtain CO2 selective MMMs from renewable or green chemistry materials. PMID:24957178

  14. Electrophoresis technology

    NASA Technical Reports Server (NTRS)

    Snyder, R. S.

    1985-01-01

    A new high resolution apparatus designed for space was built as a laboratory prototype. Using a moving wall with a low zeta potential coating, the major sources of flow distortion for an electrophoretic sample stream are removed. Highly resolved fractions, however, will only be produced in space because of the sensitivity of this chamber to buoyancy-induced convection in the laboratory. The second and third flights of the McDonnell Douglas Astronautics Corporation continuous flow electrophoresis system carried samples developed at MSFC intended to evaluate the broad capabilities of free flow electrophoresis in a reduced gravity environment. Biological model materials, hemoglobin and polystyrene latex microspheres, were selected because of their past use as electrophoresis standards and as visible markers for fluid flow due to electroosmosis, spacecraft acceleration or other factors. The dependence of the separation resolution on the properties of the sample and its suspension solution was assessed.

  15. Analysis of interspecies adherence of oral bacteria using a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis profiling

    PubMed Central

    Wang, Ren-ke; He, Xue-song; Hu, Wei; Lux, Renate; Li, Ji-yao; Zhou, Xue-dong; Shi, Wen-yuan

    2011-01-01

    Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as “bait”: Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans(S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the “bait” oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The “prey” bacterial mixtures (including known species or natural saliva samples) were added, unbound cells were washed off after the incubation period and the remaining cells were eluted using 0.2 mol·L?1 glycine. Genomic DNA was extracted, subjected to 16S rRNAPCR amplification and separation of the resulting PCR products by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F.nucleatum adhered to a variety of bacterial species including uncultivable and uncharacterized ones. This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species. PMID:21485313

  16. Expression of the AZR1 gene (ORF YGR224w), encoding a plasma membrane transporter of the major facilitator superfamily, is required for adaptation to acetic acid and resistance to azoles in Saccharomyces cerevisiae.

    PubMed

    Tenreiro, S; Rosa, P C; Viegas, C A; Sá-Correia, I

    2000-12-01

    In this work, we report results on the functional analysis of Saccharomyces cerevisiae ORF YGR224w, predicted to code for an integral membrane protein, with 14 potential transmembrane segments, belonging to the major facilitator superfamily (MFS) of transporters which are required for multiple-drug resistance (MDR). This MFS-MDR homologue is required for yeast adaptation to high stress imposed by low-chain organic acids, in particular by acetic acid, and for resistance to azoles, especially to ketoconazole and fluconazole; the encoding gene was thus named the AZR1 gene. These conclusions were based on the higher susceptibility to these compounds of an azr1Delta deletion mutant strain compared with the wild-type and on the increased resistance of both azr1Delta and wild-type strains upon increased expression of the AZR1 gene from a centromeric plasmid clone. AZR1 gene expression reduces the duration of acetic acid-induced latency, although the growth kinetics of adapted cells under acetic acid stress is apparently independent of AZR1 expression level. Fluorescence microscopy observation of the distribution of the Azr1-GFP fusion protein in yeast living cells indicated that Azr1 is a plasma membrane protein. Studies carried out to gain some understanding of how this plasma membrane putative transporter facilitates yeast adaptation to acetic acid did not implicate Azr1p in the alteration of acetic acid accumulation into the cell through the active efflux of acetate. PMID:11113970

  17. Parallel characterization of anaerobic toluene- and ethylbenzene-degrading microbial consortia by PCR-denaturing gradient gel electrophoresis, RNA-DNA membrane hybridization, and DNA microarray technology

    NASA Technical Reports Server (NTRS)

    Koizumi, Yoshikazu; Kelly, John J.; Nakagawa, Tatsunori; Urakawa, Hidetoshi; El-Fantroussi, Said; Al-Muzaini, Saleh; Fukui, Manabu; Urushigawa, Yoshikuni; Stahl, David A.

    2002-01-01

    A mesophilic toluene-degrading consortium (TDC) and an ethylbenzene-degrading consortium (EDC) were established under sulfate-reducing conditions. These consortia were first characterized by denaturing gradient gel electrophoresis (DGGE) fingerprinting of PCR-amplified 16S rRNA gene fragments, followed by sequencing. The sequences of the major bands (T-1 and E-2) belonging to TDC and EDC, respectively, were affiliated with the family Desulfobacteriaceae. Another major band from EDC (E-1) was related to an uncultured non-sulfate-reducing soil bacterium. Oligonucleotide probes specific for the 16S rRNAs of target organisms corresponding to T-1, E-1, and E-2 were designed, and hybridization conditions were optimized for two analytical formats, membrane and DNA microarray hybridization. Both formats were used to characterize the TDC and EDC, and the results of both were consistent with DGGE analysis. In order to assess the utility of the microarray format for analysis of environmental samples, oil-contaminated sediments from the coast of Kuwait were analyzed. The DNA microarray successfully detected bacterial nucleic acids from these samples, but probes targeting specific groups of sulfate-reducing bacteria did not give positive signals. The results of this study demonstrate the limitations and the potential utility of DNA microarrays for microbial community analysis.

  18. Parallel Characterization of Anaerobic Toluene- and Ethylbenzene-Degrading Microbial Consortia by PCR-Denaturing Gradient Gel Electrophoresis, RNA-DNA Membrane Hybridization, and DNA Microarray Technology

    PubMed Central

    Koizumi, Yoshikazu; Kelly, John J.; Nakagawa, Tatsunori; Urakawa, Hidetoshi; El-Fantroussi, Saïd; Al-Muzaini, Saleh; Fukui, Manabu; Urushigawa, Yoshikuni; Stahl, David A.

    2002-01-01

    A mesophilic toluene-degrading consortium (TDC) and an ethylbenzene-degrading consortium (EDC) were established under sulfate-reducing conditions. These consortia were first characterized by denaturing gradient gel electrophoresis (DGGE) fingerprinting of PCR-amplified 16S rRNA gene fragments, followed by sequencing. The sequences of the major bands (T-1 and E-2) belonging to TDC and EDC, respectively, were affiliated with the family Desulfobacteriaceae. Another major band from EDC (E-1) was related to an uncultured non-sulfate-reducing soil bacterium. Oligonucleotide probes specific for the 16S rRNAs of target organisms corresponding to T-1, E-1, and E-2 were designed, and hybridization conditions were optimized for two analytical formats, membrane and DNA microarray hybridization. Both formats were used to characterize the TDC and EDC, and the results of both were consistent with DGGE analysis. In order to assess the utility of the microarray format for analysis of environmental samples, oil-contaminated sediments from the coast of Kuwait were analyzed. The DNA microarray successfully detected bacterial nucleic acids from these samples, but probes targeting specific groups of sulfate-reducing bacteria did not give positive signals. The results of this study demonstrate the limitations and the potential utility of DNA microarrays for microbial community analysis. PMID:12088997

  19. Simulating Electrophoresis.

    ERIC Educational Resources Information Center

    Moertel, Cheryl; Frutiger, Bruce

    1996-01-01

    Describes a DNA fingerprinting simulation that uses vegetable food coloring and plastic food containers instead of DNA and expensive gel electrophoresis chambers. Allows students to decipher unknown combinations of dyes in a method similar to that used to decipher samples of DNA in DNA fingerprint techniques. (JRH)

  20. STABILITY OF MFI ZEOLITE-FILLED PDMS MEMBRANES DURING PERVAPORATIVE ETHANOL RECOVERY FROM AQUEOUS MIXTURES CONTAINING ACETIC ACID

    EPA Science Inventory

    Pervaporation is potentially a cost-effective means of recovering biofuels, such as ethanol, from biomass fermentation broths for small- to medium-scale applications (~2 - 20 million liters per year). Hydrophobic zeolite-filled polydimethylsiloxane (PDMS) membranes have been sho...

  1. Development of Low Cost Membranes (Ta, Nb & Cellulose Acetate) for H{sub 2}/CO{sub 2} Separation in WGS Reactors

    SciTech Connect

    Naidu Seetala; Upali Siriwardane

    2011-06-30

    The main aim of this work is to synthesize low temperature bimetallic nanocatalysts for Water Gas Shift reaction (WGS) for hydrogen production from CO and steam mixture; and develop low-cost metal (Nb/Ta)/ceramic membranes for H{sub 2} separation and Cellulose Acetate membranes for CO{sub 2} separation. Cu-Ni-Ce/alumina, Fe-Ni-Ce/alumina granular WGS catalysts incorporating metal oxide nanoparticles into alumina support were prepared using sol-gel/oil-drop methods. The catalysts were characterized by Powder X-ray Diffractometer (PXRD), Scanning Electron Microscope (SEM), Differential Thermal Analyzer (DTA), Thermal Gravitational Analyzer (TGA), and Brunauer, Emmett and Teller (BET) techniques. TGA shows sharp weight loss at approximately 215°C and DTA shows dehydration of metal hydroxides between 200°C and 250°C. The PXRD spectra show an increase in crystallinity as a result of heating to 1000°C, and indicating a fine dispersion of the metal oxide nanoparticles in alumina supports during the sol-gel synthesis and calcination at 450°C. BET analysis indicated a mesoporous structure of the granules with high surface area. A gas-phase dynamic flow reactor is used to optimize the reaction temperatures. A gas-phase batch reactor was used to obtain kinetic data and the parameters for maximum CO conversion. In Cu-Ni-Ce/alumina category, Cu(0%)Ni(10%)Ce(11%) was found to be the best WGS catalyst among six Low Temperature Shift (LTS) catalysts with optimum temperatures between 200-300�°C, while Ni(5%)Cu(5%)Ce(11%) was found to be the best among four High Temperature Shift (HTS) catalysts with optimum temperature between 350-400°C. In the Fe-Ni-Ce/alumina category catalysts, Fe(8%)Ni(0%)Ce(8%)/alumina and Fe(6%)Ni(2%)Ce(8%)/alumina catalysts showed optimum WGS reaction temperature below 150°C. All Ni(8-x%)Fe(x%)Ce(8%) had lower WGS reaction efficiencies compared to Ni(8-x%)Cu(x%)Ce(8%). Metal (Nb or Ta)/ceramic membranes for hydrogen separation from the WGS reaction gas products have been prepared using a) sputtering and b) aluminothermic techniques. A polyvinyl-glass permeability tester was used with a gas chromatograph (GC) for H{sub 2}/CO permeability testing. Nb films showed a higher permeability than Ta at a given disk porosity. The aluminothermically deposited membranes have higher H{sub 2} permeability compared to the sputtered films, and Nb-film coated disks showed lower H{sub 2} permeability than Ta-film. A three-stage prototype stainless steel reactor with integrated housing for 1) WGS reaction catalysts, 2) H{sub 2}/CO{sub 2} separation metal/ceramic or metal/asbestos membranes, and 3) CO/CO{sub 2} separation cellulose acetate /filter-paper membranes has been designed and tested to have capabilities to perform WGS reactions at temperatures up to 400°C and withstand gas pressures up to 15 bars. The cracking of ceramic disks and gas leaks were successfully prevented by replacing ceramic disks with asbestos sheets that can easily withstand 400°C. Kinetic studies of H{sub 2} and CO permeabilities were performed through the single and double layer Nb and Ta membranes. Cellulose acetate (CA) films with 25% triethyl citrate (TEC) as plasticizer were prepared for H{sub 2}/CO/CO{sub 2} gas separation with varying thickness of the films by acetone solutions at different concentrations and by dip-coating onto filter papers. The AFM analysis of the CA membrane showed that the uniform coating had fewer and smaller pores as the film thickness increased, and corroborated by gas permeability studies. The CO{sub 2} permeability has decreased faster than CO permeability with the CA/TEC membrane thickness, and findings support that the CA membrane could be used to entrap CO{sub 2}. Several CA/TEC membranes were also staked to increase the separation efficiency. Positron Lifetime Spectroscopy (PLS) was used to estimate the micro-porosity (pore size and concentration) and fractional free volume changes of CA/TEC films, and used to understand the variations observed in the CO{sub 2}/CO permeabilities.

  2. Fluid flow electrophoresis in space

    NASA Technical Reports Server (NTRS)

    Griffin, R. N.

    1975-01-01

    Four areas relating to free-flow electrophoresis in space were investigated. The first was the degree of improvement over earthbound operations that might be expected. The second area of investigation covered the problems in developing a flowing buffer electrophoresis apparatus. The third area of investigation was the problem of testing on the ground equipment designed for use in space. The fourth area of investigation was the improvement to be expected in space for purification of biologicals. The results of some ground-based experiments are described. Other studies included cooling requirements in space, fluid sealing techniques, and measurement of voltage drop across membranes.

  3. Agarose gel electrophoresis Solutions and reagents

    E-print Network

    Abou Elela, Sherif

    ) -Electrophoresis buffer: TAE buffer (Tris, acetate, EDTA) 50X, 1L Tris 242g Acetic acid 57,1ml EDTA 100ml ddH2O up to 1L Adjust pH to 8.3 with NaOH or acetic acid Store at RT TBE (5X) Tris 54g EDTA 4.65g Boric Acid 24g of 1M 100mM EDTA pH7.5 2ml of 0.5M 0.1% Bromophenol blue 1ml of 1% #12;0.1% xylene cyanol 200ul of 5% H

  4. Glycosaminoglycan blotting and detection after electrophoresis separation.

    PubMed

    Volpi, Nicola; Maccari, Francesca

    2015-01-01

    Separation of glycosaminoglycans (GAGs) by electrophoresis and their characterization to the microgram level are integral parts of biochemical research. Their blotting on membranes after electrophoresis offers the advantage to perform further analysis on single separated species such as identification with antibodies and/or recovery of single band. A method for the blotting and immobilizing of several nonsulfated and sulfated complex GAGs on membranes made hydrophilic and positively charged by cationic detergent after their separation by conventional agarose-gel electrophoresis is illustrated. This approach to the study of these complex macromolecules utilizes the capacity of agarose-gel electrophoresis to separate single species of polysaccharides from mixtures and the membrane technology for further preparative and analytical uses. Nitrocellulose membranes are derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of GAGs are capillary blotted after their separation in agarose-gel electrophoresis. Single purified species of variously sulfated polysaccharides are transferred on derivatized membranes with an efficiency of 100 % and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining). This enables a lower amount limit of detection of 0.1 ?g. Nonsulfated polyanions, for example hyaluronic acid (HA), may also be transferred to membranes with a limit of detection of approximately 0.1-0.5 ?g after irreversible or reversible staining. The membranes may be stained with reversible staining and the same lanes used for immunological detection or other applications. PMID:26043997

  5. Effect of K+ and Ca2+ on the indole-3-acetic acid- and fusicoccin-induced growth and membrane potential in maize coleoptile cells.

    PubMed

    Siemieniuk, Agnieszka; Karcz, Waldemar

    2015-01-01

    The role of potassium (K(+)) and calcium (Ca(2+)) in the regulation of plant growth and development is complex and needs a diverse range of physiological studies. Both elements are essential for satisfactory crop production. Here, the effects of K(+) and Ca(2+) ions on endogenous growth and growth in the presence of either indole-3-acetic acid (IAA) or fusicoccin (FC) were studied in maize (Zea mays) coleoptiles. Membrane potentials of coleoptile parenchymal cells, incubated in media containing IAA, FC and different concentrations of K(+) and Ca(2+), were also determined. Growth experiments have shown that in the absence of K(+) in the incubation medium, both endogenous and IAA- or FC-induced growth were significantly inhibited by 0.1 and 1 mM Ca(2+), respectively, while in the presence of 1 mM K(+) they were inhibited only by 1 mM Ca(2+). At 10 mM K(+), endogenous growth and growth induced by either IAA or FC did not depend on Ca(2+) concentration. TEA-Cl, a potassium channel blocker, added 1 h before IAA or FC, caused a reduction of growth by 59 or 45 %, respectively. In contrast to TEA-Cl, verapamil, the Ca(2+) channel blocker, did not affect IAA- and FC-induced growth. It was also found that in parenchymal cells of maize coleoptile segments, membrane potential (Em) was strongly affected by the medium K(+), independently of Ca(2+). However, lack of Ca(2+) in the incubation medium significantly reduced the IAA- and FC-induced membrane potential hyperpolarization. TEA-Cl applied to the control medium in the same way as in growth experiments caused Em hyperpolarization synergistic with hyperpolarization produced by IAA or FC. Verapamil did not change either the Em of parenchymal cells incubated in the control medium or the IAA- and FC-induced membrane hyperpolarization. The data presented here have been discussed considering the role of K(+) uptake channels in regulation of plant cell growth. PMID:26134122

  6. Effect of K+ and Ca2+ on the indole-3-acetic acid- and fusicoccin-induced growth and membrane potential in maize coleoptile cells

    PubMed Central

    Siemieniuk, Agnieszka; Karcz, Waldemar

    2015-01-01

    The role of potassium (K+) and calcium (Ca2+) in the regulation of plant growth and development is complex and needs a diverse range of physiological studies. Both elements are essential for satisfactory crop production. Here, the effects of K+ and Ca2+ ions on endogenous growth and growth in the presence of either indole-3-acetic acid (IAA) or fusicoccin (FC) were studied in maize (Zea mays) coleoptiles. Membrane potentials of coleoptile parenchymal cells, incubated in media containing IAA, FC and different concentrations of K+ and Ca2+, were also determined. Growth experiments have shown that in the absence of K+ in the incubation medium, both endogenous and IAA- or FC-induced growth were significantly inhibited by 0.1 and 1 mM Ca2+, respectively, while in the presence of 1 mM K+ they were inhibited only by 1 mM Ca2+. At 10 mM K+, endogenous growth and growth induced by either IAA or FC did not depend on Ca2+ concentration. TEA-Cl, a potassium channel blocker, added 1 h before IAA or FC, caused a reduction of growth by 59 or 45 %, respectively. In contrast to TEA-Cl, verapamil, the Ca2+ channel blocker, did not affect IAA- and FC-induced growth. It was also found that in parenchymal cells of maize coleoptile segments, membrane potential (Em) was strongly affected by the medium K+, independently of Ca2+. However, lack of Ca2+ in the incubation medium significantly reduced the IAA- and FC-induced membrane potential hyperpolarization. TEA-Cl applied to the control medium in the same way as in growth experiments caused Em hyperpolarization synergistic with hyperpolarization produced by IAA or FC. Verapamil did not change either the Em of parenchymal cells incubated in the control medium or the IAA- and FC-induced membrane hyperpolarization. The data presented here have been discussed considering the role of K+ uptake channels in regulation of plant cell growth. PMID:26134122

  7. Mesoxalaldehyde acetals

    SciTech Connect

    Gordeeva, G.N.; Kalashnikov, S.M.; Popov, Yu.N.; Kruglov, E.A.; Imashev, U.B.

    1987-11-10

    The treatment of methylglyoxal acetals by alkyl nitrites in the presence of the corresponding aliphatic alcohols and hydrochloric acid leads to the formation of linear mesoxalaldehyde acetals, whose structure was established by NMR spectroscopy and mass spectrometry. The major pathways for the decomposition of these molecules upon electron impact were established.

  8. Electrophoresis. [in microgravity environment

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1977-01-01

    Ground-based techniques for electrophoresis take account of the need either to circumvent the effects of gravity to prevent convection, or to use gravity for fluid stabilization through artificial density gradients. The microgravity environments of orbiting spacecraft provides a new alternative for electrophoresis by avoiding the need for either of these two approaches. The paper presents some theoretical considerations concerning electrophoresis, examines certain experimental techniques (zone and high density gel electrophoresis, isoelectric focusing and isotachophoresis), and examines the electrophoresis of living cells.

  9. Electrophoresis device

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H.; Snyder, R. S. (inventors)

    1982-01-01

    A device for separating cellular particles of a sample substance into fractionated streams of different cellular species includes a casing having a distribution chamber, a separation chamber, and a collection chamber. The electrode chambers are separated from the separation chamber interior by means of passages such that flow variations and membrane variations around the slotted portion of the electrode chamber do not enduce flow perturbations into the laminar buffer curtain flowing in the separation chamber. The cellular particles of the sample are separated under the influence of the electrical field and the separation chamber into streams of different cellular species. The streams of separated cells enter a partition array in the collection chamber where they are fractionated and collected.

  10. Phenylmercuric acetate

    Integrated Risk Information System (IRIS)

    Phenylmercuric acetate ; CASRN 62 - 38 - 4 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinog

  11. Ethyl acetate

    Integrated Risk Information System (IRIS)

    Ethyl acetate ; CASRN 141 - 78 - 6 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Eff

  12. Ammonium acetate

    Integrated Risk Information System (IRIS)

    Ammonium acetate ; CASRN 631 - 61 - 8 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic

  13. Vinyl acetate

    Integrated Risk Information System (IRIS)

    Vinyl acetate ; CASRN 108 - 05 - 4 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Eff

  14. Thallium acetate

    Integrated Risk Information System (IRIS)

    Jump to main content . Integrated Risk Information System Recent Additions | Contact Us Search : All EPA IRIS • You are here : EPA Home • Research • Environmental Assessment • IRIS • IRIS Summaries Redirect Page As of September 30 , 2009 , the assessment summary for Thallium acetate is included in t

  15. Protein electrophoresis - serum

    MedlinePLUS

    This lab test measures the types of protein in the fluid (serum) part of a blood sample. Other electrophoresis tests that measure proteins in the serum include: Immunoelectrophoresis Immunofixation Globulin electrophoresis

  16. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1980-01-01

    The following aspects of kidney cell electrophoresis are discussed: (1) the development and testing of electrophoresis solutions; (2) optimization of freezing and thawing; (3) procedures for evaluation of separated kidney cells; and (4) electrophoretic mobility characterization of kidney cells.

  17. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1979-01-01

    A kidney cell electrophoresis technique is described in four parts: (1) the development and testing of electrophoresis solutions; (2) optimization of freezing and thawing; (3) procedures for evaluation of separated kidney cells; and (4) electrophoretic mobility characteristics of kidney cells.

  18. An Economical Electrophoresis Apparatus

    ERIC Educational Resources Information Center

    Andrews, I. M.

    1975-01-01

    Describes the production of an electrophoresis apparatus from commonly discarded articles. Outlines paper and gel electrophoresis and its application to the separation of amino acids and intestinal enzymes. (GS)

  19. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P. W.

    1985-01-01

    Tasks were undertaken in support of two objectives. They are: (1) to carry out electrophoresis experiments on cells in microgravity; and (2) assess the feasibility of using purified kidney cells from embryonic kidney cultures as a source of important cell products. Investigations were carried out in the following areas: (1) ground based electrophoresis technology; (2) cell culture technology; (3) electrophoresis of cells; (4) urokinase assay research; (5) zero-g electrophoresis; and (6) flow cytometry.

  20. Histone H4 N-Terminal Acetylation in Kasumi-1 Cells Treated with Depsipeptide Determined by Acetic Acid–Urea olyacrylamide Gel Electrophoresis, Amino Acid Coded Mass Tagging, and Mass Spectrometry

    PubMed Central

    Zhang, Liwen; Su, Xiaodan; Liu, Shujun; Knapp, Amy R.; Parthun, Mark R.; Marcucci, Guido; Freitas, Michael A.

    2008-01-01

    Disrupted patterns of acetylation and deacetylation of core histones play an important role in silencing transcription of hematopoietic important genes in acute myeloid leukemia (AML). A thorough investigation of these mechanisms and the response to pharmacologic modifiers will provide a better understanding of the role of histone acetylation in leukemogenesis. We describe here an analytical approach that combines acid urea polyacrylamide gel electrophoresis (AU-PAGE), amino acid coded mass tagging (AACM), and mass spectrometry (MS) for the investigation of histone acetylation patterns. The combined approach was used to follow the dynamics of H4 acetylation in Kasumi-1 cells harboring the fusion gene AML1/ETO shown to aberrantly recruit histone deacetylases (HDACs). The histones in Kasumi-1 cells were labeled by growing the cells in media in which lysine was replaced with stable isotope-labeled lysine (Lys-D4). Labeled and unlabeled cells were treated with depsipeptide and analyzed at different time points (0, 4, 8, 12, 24, and 48 h). The cells were mixed, the histone was extracted, and acetylated H4 isoforms were separated using AU-PAGE before in-gel trypsin digestion. The digests were analyzed by MALDI-TOF MS. Peptides were identified by mass and isotope pattern. LC–MS/MS of Arg-C digests were also performed to verify the acetylation pattern for H4. The major pattern of acetylation was determined as follows: initial acetylation at K16, followed by acetylation at K12, and finally acetylation of either K8 and/or K5. PMID:17203951

  1. Kidney Cell Electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1985-01-01

    Materials and procedures for microgravity electrophoresis of living human embryonic kidney cells were evaluated, ground support in the form of analytical cell electrophoresis and flow cytometry was provided and cells returned from space flight were analyzed. Preflight culture media, electrophoresis buffer, fraction collection media, temperature profiles, and urokinase assay procedures were tested prior to flight. Electrophoretic mobility distributions of aliquots of the cell population to be fractionated in flight were obtained. The protocol established and utilized is given.

  2. Improved Electrophoresis Cell

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H.; Snyder, R. S.

    1982-01-01

    Several proposed modifications are expected to improve performance of a continous-flow electrophoresis cell. Changes would allow better control of buffer flow and would increase resolution by suppressing thermal gradients. Improved electrophoresis device would have high resolution and be easy to operate. Improvements would allow better flow control and heat dissipation.

  3. Automatic multiple applicator electrophoresis

    NASA Technical Reports Server (NTRS)

    Grunbaum, B. W.

    1977-01-01

    Easy-to-use, economical device permits electrophoresis on all known supporting media. System includes automatic multiple-sample applicator, sample holder, and electrophoresis apparatus. System has potential applicability to fields of taxonomy, immunology, and genetics. Apparatus is also used for electrofocusing.

  4. Electrophoresis of biological materials

    NASA Technical Reports Server (NTRS)

    1975-01-01

    The selection of biological products was studied for electrophoresis in space. Free flow electrophoresis, isoelectric focusing, and isotachophoresis are described. The candidates discussed include: immunoglobulins and gamma globulins; isolated islet of langerhans from pancreas; bone marrow; tumor cells; kidney cells, cryoprecipitate; and column separated cultures.

  5. Preparative electrophoresis experiment design

    NASA Technical Reports Server (NTRS)

    Thiehler, A.

    1972-01-01

    A multifaceted study supporting the NASA programs to develop a space electrophoresis capability has been conducted. The study involved principally the technique of continuous free electrophoresis. It comprised a critical review of the art, study of new techniques for enhancing resolution and stability, and construction and initial testing of a high resolution cell. The effort resulted in a significant advance in free electrophoresis technique. It has provided also a much improved base for developments exploiting the added advantages of a zero-gravity environment.

  6. Binary Oscillatory Crossflow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Molloy, Richard F.; Gallagher, Christopher T.; Leighton, David T., Jr.

    1996-01-01

    We present preliminary results of our implementation of a novel electrophoresis separation technique: Binary Oscillatory Cross flow Electrophoresis (BOCE). The technique utilizes the interaction of two driving forces, an oscillatory electric field and an oscillatory shear flow, to create an active binary filter for the separation of charged species. Analytical and numerical studies have indicated that this technique is capable of separating proteins with electrophoretic mobilities differing by less than 10%. With an experimental device containing a separation chamber 20 cm long, 5 cm wide, and 1 mm thick, an order of magnitude increase in throughput over commercially available electrophoresis devices is theoretically possible.

  7. Using capillary electrophoresis for failure analysis

    SciTech Connect

    Kelly, R.G.; Scully, H.S.; Stoner, G.E. . Center for Electrochemical Science and Engineering)

    1993-07-01

    Capillary electrophoresis (CE), an advanced solution analysis technique, can be used for failure analysis of corroded components. It has high sensitivity (concentrations as low as parts-per-trillion) and can detect quantitatively a large number of ionic species. CE determined the vapor-phase attack by organic acids, mainly acetic acid, on an electrical equipment enclosure. These acids most likely originated from the seasoning of the oak pallets used to transport the manufactured items, accumulating inside the shrink-wrap film used to bind packages to the pallet.

  8. Capillary electrophoresis-mass spectrometry of carbohydrates

    PubMed Central

    Zaia, Joseph

    2014-01-01

    The development of methods for capillary electrophoresis (CE) with on-line mass spectrometric detection (CE/MS) is driven by the need for accurate, robust and sensitive glycomics analysis for basic biomedicine, biomarker discovery, and analysis of recombinant protein therapeutics. One important capability is to profile glycan mixtures with respect to the patterns of substituents including sialic acids, acetate, sulfate, phosphate, and other groups. There is additional need for an MS-compatible separation system capable of resolving carbohydrate isomers. This review summarizes applications of CS/MS to analysis of carbohydrates, glycoproteins and glycopeptides that have appeared since 2008. Readers are referred to recent comprehensive reviews covering earlier publications. PMID:23386333

  9. Electrophoresis operations in space

    NASA Technical Reports Server (NTRS)

    Richman, D. W.

    1982-01-01

    Application of electrophoresis in space processing is described. Spaceborne experiments in areas such as biological products and FDA approved drugs are discussed. These experiments will be carried on shuttle payloads.

  10. Recent advances in preparative electrophoresis

    NASA Technical Reports Server (NTRS)

    Mosher, Richard A.; Thormann, Wolfgang; Egen, Ned B.; Couasnon, Pascal; Sammons, David W.

    1987-01-01

    Various approaches for preparative electrophoresis, and three new instruments for preparative electrophoresis are discussed. Consideration is given to isoelectric focusing, isotachophoresis, and zone electrophoresis, three gel-based electrophoresis methods. The design, functions, and performance of the Elphor VaP 21 device of Hannig (1982), the shear-stabilized BIOSTREAM separator of Thompson (1983), and the recycling isoelectric focusing device are described.

  11. Pulse Field Gel Electrophoresis.

    PubMed

    Sharma-Kuinkel, Batu K; Rude, Thomas H; Fowler, Vance G

    2016-01-01

    Pulse Field Gel Electrophoresis (PFGE) is a powerful genotyping technique used for the separation of large DNA molecules (entire genomic DNA) after digesting it with unique restriction enzymes and applying to a gel matrix under the electric field that periodically changes direction. PFGE is a variation of agarose gel electrophoresis that permits analysis of bacterial DNA fragments over an order of magnitude larger than that with conventional restriction enzyme analysis. It provides a good representation of the entire bacterial chromosome in a single gel with a highly reproducible restriction profile, providing clearly distinct and well-resolved DNA fragments. PMID:25682374

  12. Automatic multiple-sample applicator and electrophoresis apparatus

    NASA Technical Reports Server (NTRS)

    Grunbaum, B. W. (inventor)

    1977-01-01

    An apparatus for performing electrophoresis and a multiple-sample applicator is described. Electrophoresis is a physical process in which electrically charged molecules and colloidal particles, upon the application of a dc current, migrate along a gel or a membrane that is wetted with an electrolyte. A multiple-sample applicator is provided which coacts with a novel tank cover to permit an operator either to depress a single button, thus causing multiple samples to be deposited on the gel or on the membrane simultaneously, or to depress one or more sample applicators separately by means of a separate button for each applicator.

  13. DNA ELECTROPHORESIS AT SURFACES

    SciTech Connect

    RAFAILOVICH, MIRIAM; SOKOLOV, JONATHAN; GERSAPPE, DILIP

    2003-09-01

    During this year we performed two major projects: I. We developed a detailed theoretical model which complements our experiments on surface DNA electrophoresis. We found that it was possible to enhance the separation of DNA chains by imposing a chemical nanoscale pattern on the surface. This approach utilized the surface interaction effect of the DNA chains with the substrate and is a refinement to our previous method in which DNA chains were separated on homogeneous flat surfaces. By introducing the nano-patterns on the surface, the conformational changes of DNA chains of different lengths can be amplified, which results in the different friction strengths with the substrate surface. Our results also show that, when compared to the DNA electrophoresis performed on homogeneous flat surfaces, nanopatterned surfaces offer a larger window in choosing different surface interactions to achieve separation. II. In collaboration with a large international manufacturer of skin care products we also embarked on a project involving photo toxicity of titanium dioxide nanoparticles, which are a key ingredient in sunscreen and cosmetic lotions. The results clearly implicated the nanoparticles in catalyzing damage to chromosomal DNA. We then used this knowledge to develop a polymer/anti-oxidant coating which prevented the photocatalytic reaction on DNA while still retaining the UV absorptive properties of the nanoparticles. The standard gel electrophoresis was not sufficient in determining the extent of the DNA damage. The conclusions of this study were based predominantly on analysis obtained with the surface electrophoresis method.

  14. Ultra-trace determination of lead(II) in water using electrothermal atomic absorption spectrometry after preconcentration by solid-phase extraction to a small piece of cellulose acetate type membrane filter.

    PubMed

    Mizuguchi, Hitoshi; Ishida, Mirai; Takahashi, Tomohiro; Sasaki, Atsushi; Shida, Junichi

    2011-01-01

    A simple and inexpensive preconcentration technique has been developed for the ultra-trace determination of lead(II) using electrothermal atomic absorption spectrometry (ETAAS). The lead(II) complex with dicyclohexano-18-crown 6-ether (DC18C6) was extracted to a small piece of cellulose acetate-type membrane filter (2 × 5 mm) merely by vigorously eccentric stirring for 120 min under the coexistence of sodium dodecyl sulfate (SDS) at around pH 7. The extraction medium was inserted into a graphite cuvette for the determination of lead(II) by ETAAS. A linear relation was obtained for the range of 0.1-5.0 ng in 10 ml of lead(II) standard solution (r = 0.998). The detection limit was found to be 0.03 ng of lead(II) in 10 ml (0.003 µg l(-1)) of water sample. The proposed method was applied to the ultra-trace determination of lead(II) in river water, underground water, tap water, and snow fall samples. PMID:21233566

  15. Preparative electrophoresis for space

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1988-01-01

    A premise of continuous flow electrophoresis is that removal of buoyance-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chamber are used, distortion of the injected sample stream due to electrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field were not considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

  16. Electrophoresis experiments in microgravity

    NASA Technical Reports Server (NTRS)

    Snyder, Robert S.; Rhodes, Percy H.

    1991-01-01

    The use of the microgravity environment to separate and purify biological cells and proteins has been a major activity since the beginning of the NASA Microgravity Science and Applications program. Purified populations of cells are needed for research, transplantation and analysis of specific cell constituents. Protein purification is a necessary step in research areas such as genetic engineering where the new protein has to be separated from the variety of other proteins synthesized from the microorganism. Sufficient data are available from the results of past electrophoresis experiments in space to show that these experiments were designed with incomplete knowledge of the fluid dynamics of the process including electrohydrodynamics. However, electrophoresis is still an important separation tool in the laboratory and thermal convection does limit its performance. Thus, there is a justification for electrophoresis but the emphasis of future space experiments must be directed toward basic research with model experiments to understand the microgravity environment and fluid analysis to test the basic principles of the process.

  17. Preparative electrophoresis for space

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1987-01-01

    A premise of continuous flow electrophoresis is that removal of buoyancy-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chambers are used, distortion of the injected sample stream due to electrohydrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field have not been considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

  18. A unique enzyme of acetic acid bacteria, PQQ-dependent alcohol dehydrogenase, is also present in Frateuria aurantia.

    PubMed

    Tr?ek, Janja; Matsushita, Kazunobu

    2013-08-01

    A membrane-bound, pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase (ADH) was purified from Frateuria aurantia LMG 1558(T). Although F. aurantia belongs to a group of ?-Proteobacteria, the characteristics of its PQQ-ADH were similar to the enzyme characteristics of the typical high-acetic acid-resistant bacterium Gluconacetobacter europaeus from the group of ?-Proteobacteria. The PQQ-dependent ADH was solubilized from the membranes and purified after anionic, cationic, and affinity chromatography with specific activity of 117 U/mg. The purified enzyme was estimated to be composed of two subunits of ca. 72 and 45 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The purified enzyme had maximum activity at pH 4.5 and showed the highest substrate specificity to ethanol, isoamyl alcohol, 1-butanol, and 1-propanol. The deduced sequences of cloned genes adhA and adhB encoding subunits I and II of PQQ-ADH showed 80 % amino acid (AA) identity to AdhA and 68 % AA identity to AdhB of Ga. europaeus V3 (LMG 18494). Because of the high similarity between genes encoding subunits I and II of PQQ-ADH and its homologous genes found in a distantly related taxonomic group of acetic acid bacteria, the results suggest the possibility of horizontal gene transfer between these two groups of genera. PMID:23760531

  19. Possibility of Microchip Electrophoresis for Biological Application

    NASA Astrophysics Data System (ADS)

    Kataoka, Masatoshi; Kido, Jun-Ichi; Shinohara, Yasuo

    Microchip electrophoresis has recently attracted much attention in the field of nuclear acid analysis due to its high efficiency, ease of operation, low consumption of samples and reagents, and relatively low costs. Nucleic acid fragments are separated by capillary electrophoresis in a chip with microfabricated channels, with automated detection as well as on-line data evaluation. Microfabricated devices are forecast to be fundamental to the postgenome era, especially in the field of genetics and medicine. However, although there are many reports of the use of these instruments to evaluate standard DNA, DNA ladders, PCR products, and commercially available plasmid digests, little information is available their use with biological material. In this report, we showed the accuracy of sizing and quantification of endonuclease-digested plasmid DNA. We also showed the feasibility of on-microchip endonuclease treatment of plasmid DNA and sequential analysis as an additional application for DNA analysis. Furthermore, to evaluate the possibility of microchip electrophoresis for biological application, the results of the examination of blood sugar in human plasma and mitochondrial membrane potential were shown.

  20. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Chang, H.T.; Fung, E.N.; Li, Q.; Lu, X.

    1996-12-10

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  1. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Li, Q.; Lu, X.

    1998-04-21

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  2. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S. (Ames, IA); Li, Qingbo (Ames, IA); Lu, Xiandan (Ames, IA)

    1998-04-21

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  3. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S. (Ames, IA); Chang, Huan-Tsang (Silver Spring, MD); Fung, Eliza N. (Ames, IA); Li, Qingbo (Ames, IA); Lu, Xiandan (Ames, IA)

    1996-12-10

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  4. Preparation of vinyl acetate

    DOEpatents

    Tustin, G.C.; Zoeller, J.R.; Depew, L.S.

    1998-03-24

    This invention pertains to the preparation of vinyl acetate by contacting a mixture of hydrogen and ketene with a heterogeneous catalyst containing a transition metal to produce acetaldehyde, which is then reacted with ketene in the presence of an acid catalyst to produce vinyl acetate.

  5. Kidney cell electrophoresis, continuing task

    NASA Technical Reports Server (NTRS)

    Todd, P. W.

    1985-01-01

    Materials and procedures for microgravity electrophoresis of living human embryonic kidney cells were evaluated to provide ground support in the form of analytical cell electrophoresis and flow cytometry. Preflight culture media, electrophoresis buffer, fraction collection media, temperature profiles, and urokinase assay procedures were tested prior to flight. Electrophoretic mobility distributions of aliquots of the cell population to be fractionated in flight were obtained. Cells were prepared in suspension prior to flight in electrophoresis buffer and 10% calf serum. Electrophoretic separation proceeded in electrophoresis buffer without serum in the Continuous Flow Electrophoretic Separator, and fractions were collected into sample bags containing culture medium and concentrated serum. Fractions that yielded enough progeny cells were analyzed for morphology and electrophoretic mobility distributions. It is noted that the lowest mobility fraction studied produced higher mobility progeny while the other fractions produced progeny cells with mobilities related to the fractions from which they were collected.

  6. Electrophoresis demonstration on Apollo 16

    NASA Technical Reports Server (NTRS)

    Snyder, R. S.

    1972-01-01

    Free fluid electrophoresis, a process used to separate particulate species according to surface charge, size, or shape was suggested as a promising technique to utilize the near zero gravity condition of space. Fluid electrophoresis on earth is disturbed by gravity-induced thermal convection and sedimentation. An apparatus was developed to demonstrate the principle and possible problems of electrophoresis on Apollo 14 and the separation boundary between red and blue dye was photographed in space. The basic operating elements of the Apollo 14 unit were used for a second flight demonstration on Apollo 16. Polystyrene latex particles of two different sizes were used to simulate the electrophoresis of large biological particles. The particle bands in space were extremely stable compared to ground operation because convection in the fluid was negligible. Electrophoresis of the polystyrene latex particle groups according to size was accomplished although electro-osmosis in the flight apparatus prevented the clear separation of two particle bands.

  7. Electrophoresis experiment for space

    NASA Technical Reports Server (NTRS)

    Vanderhoff, J. W.; Micale, F. J.

    1976-01-01

    The Apollo 16 electrophoresis experiment was analyzed, demonstrating that the separation of the two different-size monodisperse latexes did indeed take place, but that the separation was obscured by the pronounced electroosmotic flow of the liquid medium. The results of this experiment, however, were dramatic since it is impossible to carry out a similar separation on earth. It can be stated unequivocally from this experiment that any electrophoretic separation will be enhanced under microgravity conditions. The only question is the degree of this enhancement, which can be expected to vary from one experimental technique to another. The low-electroosmotic-mobility coating (Z6040-MC) developed under this program was found to be suitable for a free-fluid electrophoretic separation such as the experiment designed for the ASTP flight. The problem with this coating, however, is that its permanency is limited because of the slow desorption of the methylcellulose from the coated surface.

  8. Static continuous electrophoresis device

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H. (inventor)

    1982-01-01

    An apparatus is disclosed for carrying out a moving wall type electrophoresis process for separation of cellular particles. The apparatus includes a water-tight housing containing an electrolytic buffer solution. A separation chamber in the housing is defined by spaced opposed moving walls and spaced opposed side walls. Substrate assemblies, which support the moving wall include vacuum ports for positively sealing the moving walls against the substrate walls. Several suction conduits communicate with the suction ports and are arranged in the form of valleys in a grid plate. The raised land portion of the grid plat supports the substrate walls against deformation inwardly under suction. A cooling chamber is carried on the back side of plate. The apparatus also has tensioner means including roller and adjustment screws for maintaining the belts in position and a drive arrangement including an electric motor with a gear affixed to its output shaft. Electrode assemblies are disposed to provide the required electric field.

  9. The Acetate Switch

    PubMed Central

    Wolfe, Alan J.

    2005-01-01

    To succeed, many cells must alternate between life-styles that permit rapid growth in the presence of abundant nutrients and ones that enhance survival in the absence of those nutrients. One such change in life-style, the “acetate switch,” occurs as cells deplete their environment of acetate-producing carbon sources and begin to rely on their ability to scavenge for acetate. This review explains why, when, and how cells excrete or dissimilate acetate. The central components of the “switch” (phosphotransacetylase [PTA], acetate kinase [ACK], and AMP-forming acetyl coenzyme A synthetase [AMP-ACS]) and the behavior of cells that lack these components are introduced. Acetyl phosphate (acetyl?P), the high-energy intermediate of acetate dissimilation, is discussed, and conditions that influence its intracellular concentration are described. Evidence is provided that acetyl?P influences cellular processes from organelle biogenesis to cell cycle regulation and from biofilm development to pathogenesis. The merits of each mechanism proposed to explain the interaction of acetyl?P with two-component signal transduction pathways are addressed. A short list of enzymes that generate acetyl?P by PTA-ACKA-independent mechanisms is introduced and discussed briefly. Attention is then directed to the mechanisms used by cells to “flip the switch,” the induction and activation of the acetate-scavenging AMP-ACS. First, evidence is presented that nucleoid proteins orchestrate a progression of distinct nucleoprotein complexes to ensure proper transcription of its gene. Next, the way in which cells regulate AMP-ACS activity through reversible acetylation is described. Finally, the “acetate switch” as it exists in selected eubacteria, archaea, and eukaryotes, including humans, is described. PMID:15755952

  10. Comparison of the phosphorylation events in membranes prepared from proliferating versus quiescent endothelial cells

    SciTech Connect

    Kazlauskas, A.; DiColeto, P.E.

    1986-05-01

    Little is known of the intracellular events which regulate the proliferation of endothelial cells (EC). Triton-solubilized membranes from proliferating (sparse) and quiescent (confluent) EC were incubated at pH 6.5 in the presence of divalent cations and (/sup 32/P)ATP. Membrane proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. The overall kinase activity per mg protein was slightly greater in membranes prepared from proliferating versus quiescent cells. They found four proteins labeled in sparse cells to a dramatically greater extent having the following approximate molecular masses: 180, 100, 97 and 55 kilodalton (kd). The first two phosphoproteins were phosphorylated on serine residues exclusively; the 97 kd phosphoprotein contained 39% phosphoserine (p-ser) and 61% phosphothreonine (p-thr); and the 55 kd phosphoprotein contained 62% p-ser, 16% p-thr, and 22% phosphotyrosine (p-tyr). The kinases acting on all four phosphoproteins were independent of Ca/sup 2 +/, cAMP, cGMP, or phorbol 12-myristate 13-acetate. The observed differences in phosphorylation events between sparse and confluent membranes occurred in membranes from two EC lines - pig aortic and bovine aortic - but were not apparent in membranes prepared from human foreskin fibroblasts or 3T3 cells. Sparse endothelial cells made quiescent by serum deprivation were found to resemble confluent cells in the kinase activity; therefore, the enhanced kinase activity in sparse membranes may be growth dependent.

  11. Automated dual capillary electrophoresis system with hydrodynamic injection for the concurrent determination of cations and anions.

    PubMed

    Pham, Thi Thanh Thuy; Mai, Thanh Duc; Nguyen, Thanh Dam; Sáiz, Jorge; Pham, Hung Viet; Hauser, Peter C

    2014-09-01

    The capillary electrophoresis instrument developed for the concurrent determination of cations and anions features two separate capillaries and individual detectors to allow independent optimization for each group of ions. The capillaries are joined in a common injector block. The sample is drawn into the injector with a small membrane pump and automated simultaneous injection into both capillaries is achieved by pressurization of the fluid with compressed air. Flushing of the injector and of the capillaries with the background electrolyte is also carried out automatically by the same means. The buffer consisted of 12mM histidine and 2mM 18-crown-6 adjusted to pH 4 with acetic acid and was suitable for the contactless conductivity detection employed. The system was optimized for the determination of cationic NH4(+) and anionic NO3(-) and NO2(-), and linear calibration curves from about 20?M up to about 1.5mM were obtained for these ions. In a test run over 8h, the reproducibility for the peak areas was within ±7%. For demonstration, the instrument was successfully applied to the concurrent monitoring of the concentrations of the three ions during the biological removal of ammonium from contaminated groundwater in a sequencing batch reactor, where NO3(-) and NO2(-) are formed as intermediate products. PMID:25109864

  12. Binary Oscillatory Crossflow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Molloy, Richard F.; Gallagher, Christopher T.; Leighton, David T., Jr.

    1997-01-01

    Electrophoresis has long been recognized as an effective analytic technique for the separation of proteins and other charged species, however attempts at scaling up to accommodate commercial volumes have met with limited success. In this report we describe a novel electrophoretic separation technique - Binary Oscillatory Crossflow Electrophoresis (BOCE). Numerical simulations indicate that the technique has the potential for preparative scale throughputs with high resolution, while simultaneously avoiding many problems common to conventional electrophoresis. The technique utilizes the interaction of an oscillatory electric field and a transverse oscillatory shear flow to create an active binary filter for the separation of charged protein species. An oscillatory electric field is applied across the narrow gap of a rectangular channel inducing a periodic motion of charged protein species. The amplitude of this motion depends on the dimensionless electrophoretic mobility, alpha = E(sub o)mu/(omega)d, where E(sub o) is the amplitude of the electric field oscillations, mu is the dimensional mobility, omega is the angular frequency of oscillation and d is the channel gap width. An oscillatory shear flow is induced along the length of the channel resulting in the separation of species with different mobilities. We present a model that predicts the oscillatory behavior of charged species and allows estimation of both the magnitude of the induced convective velocity and the effective diffusivity as a function of a in infinitely long channels. Numerical results indicate that in addition to the mobility dependence, the steady state behavior of solute species may be strongly affected by oscillating fluid into and out of the active electric field region at the ends of the cell. The effect is most pronounced using time dependent shear flows of the same frequency (cos((omega)t)) flow mode) as the electric field oscillations. Under such conditions, experiments indicate that solute is drawn into the cell from reservoirs at both ends of the cell leading to a large mass build up. As a consequence, any initially induced mass flux will vanish after short times. This effect was not captured by the infinite channel model and hence numerical and experimental results deviated significantly. The revised model including finite cell lengths and reservoir volumes allowed quantitative predictions of the time history of the concentration profile throughout the system. This latter model accurately describes the fluxes observed for both oscillatory flow modes in experiments using single protein species. Based on the results obtained from research funded under NASA grant NAG-8-1080.S, we conclude that binary separations are not possible using purely oscillatory flow modes because of end effects associated with the cos((omega)t) mode. Our research shows, however, that a combination of cos(2(omega)t) and steady flow should lead to efficient separation free of end effects. This possibility is currently under investigation.

  13. Biomedical applications of capillary electrophoresis

    NASA Astrophysics Data System (ADS)

    Kartsova, L. A.; Bessonova, E. A.

    2015-08-01

    The review deals with modern analytical approaches used in capillary electrophoresis for solving medical and biological problems: search for biomarkers of various diseases and rapid diagnosis based on characteristic profiles of biologically active compounds by capillary electrophoresis with mass spectrometric detection; monitoring of the residual drugs in biological fluids for evaluating the efficiency of drug therapy; testing of the enantiomeric purity of pharmaceutical products; the use of novel materials as components of stationary and pseudo-stationary phases in capillary electrophoresis and capillary electrochromatography to increase the selectivity of separation of components of complex matrices; and identification of various on-line preconcentration techniques to reduce the detection limits of biologically active analytes. A topical trend in capillary electrophoresis required in clinical practice, viz., the design of microfluidic systems, is discussed. The bibliography includes 173 references.

  14. Copolymers For Capillary Gel Electrophoresis

    DOEpatents

    Liu, Changsheng (State College, PA); Li, Qingbo (State College, PA)

    2005-08-09

    This invention relates to an electrophoresis separation medium having a gel matrix of at least one random, linear copolymer comprising a primary comonomer and at least one secondary comonomer, wherein the comonomers are randomly distributed along the copolymer chain. The primary comonomer is an acrylamide or an acrylamide derivative that provides the primary physical, chemical, and sieving properties of the gel matrix. The at least one secondary comonomer imparts an inherent physical, chemical, or sieving property to the copolymer chain. The primary and secondary comonomers are present in a ratio sufficient to induce desired properties that optimize electrophoresis performance. The invention also relates to a method of separating a mixture of biological molecules using this gel matrix, a method of preparing the novel electrophoresis separation medium, and a capillary tube filled with the electrophoresis separation medium.

  15. DNA typing by capillary electrophoresis

    SciTech Connect

    Zhang, N.

    1997-10-08

    Capillary electrophoresis is becoming more and more important in nucleic acid analysis including DNA sequencing, typing and disease gene measurements. This work summarized the background of DNA typing. The recent development of capillary electrophoresis was also discussed. The second part of the thesis showed the principle of DNA typing based on using the allelic ladder as the absolute standard ladder in capillary electrophoresis system. Future work will be focused on demonstrating DNA typing on multiplex loci and examples of disease diagnosis in the on-line format of PCR-CE. Also capillary array electrophoresis system should allow high throughput, fast speed DNA typing. Only the introduction and conclusions for this report are available here. A reprint was removed for separate processing.

  16. Rapid analysis of atorvastatin calcium using capillary electrophoresis and microchip electrophoresis.

    PubMed

    Guihen, Elizabeth; Sisk, Garry D; Scully, Norma M; Glennon, Jeremy D

    2006-06-01

    In this work, a capillary electrophoretic method for the rapid quantitation of atorvastatin (AT) in a lipitor tablet was investigated and developed. Method development included studies of the effect of applied potential, buffer concentration, buffer pH, and hydrodynamic injection time on the electrophoretic separation. The method was validated with regard to linearity, precision, specificity, LOD, and LOQ. The optimum electrophoretic separation conditions were 25 mM sodium acetate buffer at pH 6, with a separation voltage of 25 kV using a 50 microm capillary of 33 cm total length. Sodium diclofenac was used as an internal standard. Analysis of AT in a commercial lipitor tablet by electrophoresis gave quite high efficiency, coupled with an analysis time of less than 1.2 min in comparison to LC. Once the separation was optimized on capillary, it was further miniaturized to a microchip platform, with linear imaging UV detection using microchip electrophoresis (MCE). Linear imaging UV detection allowed for real-time monitoring of the analyte movement on chip, so that the optimum separation time could be easily determined. This microchip electrophoretic method was compared to the CE method with regard to speed, efficiency, precision, and LOD. This work represents the most rapid and first reported analysis of AT using MCE. PMID:16786480

  17. Frequently made mistakes in electrophoresis.

    PubMed

    Westermeier, Reiner

    2007-09-01

    In spite of text books, instrument manuals, product instructions, and web tutorials there are a number of erroneous protocols around, which lead repeatedly to issues during electrophoresis runs and to inadequate results. The relatively low resolution and short running time of miniformat systems often conceals these issues. However, in high-resolution 2-D electrophoresis in large format gels, one of the most important separation methods in Proteomics, the consequences of these mistakes become more obvious. PMID:17893853

  18. Electrophoretic Characterization of a Detergent-Treated Plasma Membrane Fraction from Corn Roots 1

    PubMed Central

    Gallagher, Sean R.; Leonard, Robert T.

    1987-01-01

    Experiments were conducted to determine conditions essential for electrophoretic characterization of a detergent-extracted plasma membrane fraction from corn (Zea mays L.) roots. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) initially gave poor resolution of polypeptides in the plasma membrane fraction and, upon detergent treatment for purification of the proton-pumping adenosine triphosphatase (ATPase), showed no enrichment for a 100 kilodalton catalytic subunit characteristic of the ATPase. In contrast to SDS-PAGE, phenol urea acetic acid (PAU)-PAGE clearly resolved two polypeptides in the 100 kilodalton region that were enriched during detergent treatment and indicated at least one polypeptide forms a phosphorylated intermediate characteristic of the ATPase. Problems with SDS-PAGE were found to be caused, in part, by a combination of endogenous proteases and heat-induced aggregation of high molecular weight proteins. The usually standard procedure of boiling the sample prior to SDS-PAGE caused the aggregation of the 100 kilodalton polypeptides. By controlling for proteases using chymostatin and/or phenylmethane sulfonyl floride, and not boiling the sample prior to electrophoresis, two polypeptides were clearly resolved by SDS-PAGE in the 100 kilodalton region of Triton X-114-extracted membranes from corn, oat, barley, and tomato. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:16665234

  19. Antibody enhancement of free-flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Cohly, H. H. P.; Morrison, Dennis R.; Atassi, M. Zouhair

    1988-01-01

    Specific T cell clones and antibodies (ABs) were developed to study the efficiency of purifying closely associated T cells using Continuous Flow Electrophoresis System. Enhanced separation is accomplished by tagging cells first with ABs directed against the antigenic determinants on the cell surface and then with ABs against the Fc portion of the first AB. This second AB protrudes sufficiently beyond the cell membrane and glycocalyx to become the major overall cell surface potential determinant and thus causes a reduction of electrophoretic mobility. This project was divided into three phases. Phase one included development of specific T cell clones and separation of these specific clones. Phase two extends these principles to the separation of T cells from spleen cells and immunized lymph node cells. Phase three applies this double antibody technique to the separation of T cytotoxic cells from bone marrow.

  20. PNEUMATIC MICROVALVE FOR ELECTROKINETIC SAMPLE PRECONCENTRATION AND CAPILLARY ELECTROPHORESIS INJECTION

    SciTech Connect

    Cong, Yongzheng; Rausch, Sarah J.; Geng, Tao; Jambovane, Sachin R.; Kelly, Ryan T.

    2014-10-27

    Here we show that a closed pneumatic microvalve on a PDMS chip can serve as a semipermeable membrane under an applied potential, enabling current to pass through while blocking the passage of charged analytes. Enrichment of both anionic and cationic species has been demonstrated, and concentration factors of ~70 have been achieved in just 8 s. Once analytes are concentrated, the valve is briefly opened and the sample is hydrodynamically injected onto an integrated microchip or capillary electrophoresis (CE) column. In contrast to existing preconcentration approaches, the membrane-based method described here enables both rapid analyte concentration as well as high resolution separations.

  1. Fragrance material review on ?-methylbenzyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of ?-methylbenzyl acetate when used as a fragrance ingredient is presented. ?-Methylbenzyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for ?-methylbenzyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, skin sensitization, elicitation, and repeated dose data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22406576

  2. Fragrance material review on benzyl acetate.

    PubMed

    McGinty, D; Vitale, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of benzyl acetate when used as a fragrance ingredient is presented. Benzyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for benzyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, skin sensitization, elicitation, phototoxicity, toxicokinetics, repeated dose, reproductive toxicity, genotoxicity, or carcinogenicity data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Refer Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22387848

  3. Fragrance material review on 2-phenylpropyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 2-phenylpropyl acetate when used as a fragrance ingredient is presented. 2-Phenylpropyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 2-phenylpropyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, and skin sensitization data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22421639

  4. Fragrance material review on phenethyl acetate.

    PubMed

    McGinty, D; Vitale, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of phenethyl acetate when used as a fragrance ingredient is presented. Phenethyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for phenethyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, skin sensitization, elicitation, toxicokinetics, repeated dose, genotoxicity, and carcinogenicity data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414644

  5. Fragrance material review on piperonyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of piperonyl acetate when used as a fragrance ingredient is presented. Piperonyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for piperonyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, skin sensitization, toxicokinetics, and genotoxicity data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22445840

  6. Contactless conductivity detector for microchip capillary electrophoresis

    NASA Technical Reports Server (NTRS)

    Pumera, Martin; Wang, Joseph; Opekar, Frantisek; Jelinek, Ivan; Feldman, Jason; Lowe, Holger; Hardt, Steffen; Svehla, D. (Principal Investigator)

    2002-01-01

    A microfabricated electrophoresis chip with an integrated contactless conductivity detection system is described. The new contactless conductivity microchip detector is based on placing two planar sensing aluminum film electrodes on the outer side of a poly(methyl methacrylate) (PMMA) microchip (without contacting the solution) and measuring the impedance of the solution in the separation channel. The contactless route obviates problems (e.g., fouling, unwanted reactions) associated with the electrode-solution contact, offers isolation of the detection system from high separation fields, does not compromise the separation efficiency, and greatly simplifies the detector fabrication. Relevant experimental variables, such as the frequency and amplitude of the applied ac voltage or the separation voltage, were examined and optimized. The detector performance was illustrated by the separation of potassium, sodium, barium, and lithium cations and the chloride, sulfate, fluoride, acetate, and phosphate anions. The response was linear (over the 20 microM-7 mM range) and reproducible (RSD = 3.4-4.9%; n = 10), with detection limits of 2.8 and 6.4 microM (for potassium and chloride, respectively). The advantages associated with the contactless conductivity detection, along with the low cost of the integrated PMMA chip/detection system, should enhance the power and scope of microfluidic analytical devices.

  7. Electrophoresis as a management tool

    USGS Publications Warehouse

    Morgan, R.P., II; Chapman, J.A.; Noe, L.A.; Henny, C.J.

    1974-01-01

    The theme of this 1974 Northeast Fish and Wildlife Conference is 'A New Era'. Indeed, it is a new era for improved techniques to assist in management of our fish and wildlife resources for the maximum benefit of all. In some cases, the new techniques are primarily used in research.on fish and wildlife, and the results from the research are used to aid management and enforcement agencies in the decision-making process. One of the newer techniques that is being applied to problems in fisheries and wildlife is electrophoresis. In this paper, we review briefly the techniques of electrophoresis and illustrate research problems in wildlife and fisheries where the use of electrophoresis is now assisting or may potentially aid in management decisions.

  8. Outer membrane vesicles of the VA-MENGOC-BC vaccine against serogroup B of Neisseria meningitidis: Analysis of protein components by two-dimensional gel electrophoresis and mass spectrometry.

    PubMed

    Uli, Liliam; Castellanos-Serra, Lila; Betancourt, Lazaro; Domínguez, Francisco; Barberá, Ramón; Sotolongo, Franklin; Guillén, Gerardo; Pajón Feyt, Rolando

    2006-06-01

    Neisseria meningitidis is a Gram-negative bacterium responsible for significant mortality worldwide. While effective polysaccharides-based vaccines exist against serogroups A, C, W135, and Y, no similar vaccine is suitable for children under 4 years against disease caused by serogroup B strains. Therefore, major vaccine efforts against this serogroup are based on outer membrane vesicles (OMVs), containing major outer membrane proteins. The OMV-based vaccine produced by the Finlay Institute in Cuba (VA-MENGOC-BC) contributed to the rapid decline of the epidemic in this Caribbean island. While the content of major proteins in this vaccine has been discussed, no detailed work of an outer membrane proteomic map of this, or any other, commercially available OMV-derived product has been published so far. Since OMVs exhibit a large bias toward a few major proteins and usually contain a high content of lipids, establishing the adequate conditions for high resolution, 2-DE of this kind of preparation was definitely a technical challenge. In this work, 2-DE and MS have been used to generate a proteomic map of this product, detailing the presence of 31 different proteins, and it allows the identification of new putative protective protein components it contains. PMID:16673438

  9. Vinegar as a burn-down herbicide: Acetic acid concentrations, application volumes, and adjuvants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acetic acid acts as a contact herbicide, injuring and killing plants by first destroying the cell membranes, which causes the rapid desiccation of the plant tissues. Vinegars with acetic acid concentrations of 11% or greater can burn the skin and cause serious to severe eye injury, including blindn...

  10. Fast separation and analysis of reduced monoclonal antibodies with capillary zone electrophoresis coupled to mass spectrometry.

    PubMed

    Zhao, Yimeng; Sun, Liangliang; Knierman, Michael D; Dovichi, Norman J

    2016-02-01

    Capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE-ESI-MS) was used for analysis of reduced antibodies. We first developed a simple protocol to condition commercial linear-polyacrylamide coated capillaries for use in top-down proteomics. We then suspended reduced antibodies in a solution of 35% acetic acid, 50% acetonitrile in water. Heavy and light chains were baseline resolved within 10min and with 3-30µg/mL detection limits using a 0.1% aqueous formic acid background electrolyte. Quintuplicate runs of a two-antibody mixture produced relative standard deviations of ?1% in migration time and 10% in peak amplitudes. Resolution was further improved for the two-antibody mixture by using 5% acetic acid as the background electrolyte, highlighting the potential of capillary electrophoresis-mass spectrometry for analysis of antibody mixtures. PMID:26653481

  11. Techniques For Focusing In Zone Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Twitty, Garland E.; Sammons, David W.

    1994-01-01

    In two techniques for focusing in zone electrophoresis, force of applied electrical field in each charged particle balanced by restoring force of electro-osmosis. Two techniques: velocity-gradient focusing (VGF), suitable for rectangular electrophoresis chambers; and field-gradient focusing (FGF), suitable for step-shaped electrophoresis chambers.

  12. Bicarbonate versus acetate hemodialysis in ventilated patients.

    PubMed

    van Geelen, J A; Woittiez, A J; Schalekamp, M A

    1987-09-01

    Hemodynamic tolerance to bicarbonate versus acetate hemodialysis was studied in seven ventilated, critically ill patients, suffering from acute renal failure. Both kinds of hemodialysis were carried out with a recirculating dialysate delivery system and a relatively low blood flow (180 ml/min). Each patient underwent two hemodialysis procedures, one with bicarbonate and one with acetate, lasting for four hours. Ultrafiltration rates were kept below 250 ml/h and only biocompatible membranes with a relatively small surface area (Biospal 2400, Hospal, France) were used. Despite the mild hemodialysis conditions, hypotensive episodes with a mean blood pressure below 70 mmHg were observed in 3 out of 7 bicarbonate sessions and 4 out of 7 acetate sessions. Thus, we could not demonstrate a hemodynamic advantage of bicarbonate hemodialysis in this group of ventilated patients. This contrasts with other studies conducted in non-ventilated patients. Prevention of hypoxemia by mechanical ventilation and control of vascular tone by the use of vasoactive drugs may be of more clinical relevance than the kind of hemodialysis procedure that is used. PMID:3117466

  13. Membranes and Films from Polymers.

    ERIC Educational Resources Information Center

    Blumberg, Avrom A.

    1986-01-01

    Provides background information on polymeric films and membranes including production methods, special industrial and medical applications, laboratory preparation, and an experimental investigation of a porous cellulose acetate membrane. Presents a demonstration to distinguish between high- and low-density polyethylene. (JM)

  14. Capillary electrophoresis systems and methods

    DOEpatents

    Dorairaj, Rathissh (Hillsboro, OR); Keynton, Robert S. (Louisville, KY); Roussel, Thomas J. (Louisville, KY); Crain, Mark M. (Georgetown, IN); Jackson, Douglas J. (New Albany, IN); Walsh, Kevin M. (Louisville, KY); Naber, John F. (Goshen, KY); Baldwin, Richard P. (Louisville, KY); Franco, Danielle B. (Mount Washington, KY)

    2011-08-02

    An embodiment of the invention is directed to a capillary electrophoresis apparatus comprising a plurality of separation micro-channels. A sample loading channel communicates with each of the plurality of separation channels. A driver circuit comprising a plurality of electrodes is configured to induce an electric field across each of the plurality of separation channels sufficient to cause analytes in the samples to migrate along each of the channels. The system further comprises a plurality of detectors configured to detect the analytes.

  15. Enhancing Centrifugal Separation With Electrophoresis

    NASA Technical Reports Server (NTRS)

    Herrmann, F. T.

    1986-01-01

    Separation of biological cells by coil-planet centrifuge enhanced by electrophoresis. By itself, coil-planet centrifuge offers relatively gentle method of separating cells under low centrifugal force in physiological medium that keeps cells alive. With addition of voltage gradient to separation column of centrifuge, separation still gentle but faster and more complete. Since separation apparatus contains no rotary seal, probability of leakage, contamination, corrosion, and short circuits reduced.

  16. Boswellic acid acetate induces apoptosis through caspase-mediated pathways in myeloid leukemia cells.

    PubMed

    Xia, Lijuan; Chen, Duo; Han, Rui; Fang, Qicheng; Waxman, Samuel; Jing, Yongkui

    2005-03-01

    The mechanism of the cytotoxic effect of boswellic acid acetate, a 1:1 mixture of alpha-boswellic acid acetate and beta-boswellic acid acetate, isolated from Boswellia carterri Birdw on myeloid leukemia cells was investigated in six human myeloid leukemia cell lines (NB4, SKNO-1, K562, U937, ML-1, and HL-60 cells). Morphologic and DNA fragmentation assays indicated that the cytotoxic effect of boswellic acid acetate was mediated by induction of apoptosis. More than 50% of the cells underwent apoptosis after treatment with 20 mug/mL boswellic acid for 24 hours. This apoptotic process was p53 independent. The levels of apoptosis-related proteins Bcl-2, Bax, and Bcl-XL were not modulated by boswellic acid acetate. Boswellic acid acetate induced Bid cleavage and decreased mitochondrial membrane potential without production of hydrogen peroxide. A general caspase inhibitor (Z-VAD-FMK) and a specific caspase-8 inhibitor II (Z-IETD-FMK) blocked boswellic acid acetate-induced apoptosis. The mRNAs of death receptors 4 and 5 (DR4 and DR5) were induced in leukemia cells undergoing apoptosis after boswellic acid acetate treatment. These data taken together suggest that boswellic acid acetate induces myeloid leukemia cell apoptosis through activation of caspase-8 by induced expression of DR4 and DR5, and that the activated caspase-8 either directly activates caspase-3 by cleavage or indirectly by cleaving Bid, which in turn decreases mitochondria membrane potential. PMID:15767547

  17. Scaleable production and separation of fermentation-derived acetic acid. Final CRADA report.

    SciTech Connect

    Snyder, S. W.; Energy Systems

    2010-02-08

    Half of U.S. acetic acid production is used in manufacturing vinyl acetate monomer (VAM) and is economical only in very large production plants. Nearly 80% of the VAM is produced by methanol carbonylation, which requires high temperatures and exotic construction materials and is energy intensive. Fermentation-derived acetic acid production allows for small-scale production at low temperatures, significantly reducing the energy requirement of the process. The goal of the project is to develop a scaleable production and separation process for fermentation-derived acetic acid. Synthesis gas (syngas) will be fermented to acetic acid, and the fermentation broth will be continuously neutralized with ammonia. The acetic acid product will be recovered from the ammonium acid broth using vapor-based membrane separation technology. The process is summarized in Figure 1. The two technical challenges to success are selecting and developing (1) microbial strains that efficiently ferment syngas to acetic acid in high salt environments and (2) membranes that efficiently separate ammonia from the acetic acid/water mixture and are stable at high enough temperature to facilitate high thermal cracking of the ammonium acetate salt. Fermentation - Microbial strains were procured from a variety of public culture collections (Table 1). Strains were incubated and grown in the presence of the ammonium acetate product and the fastest growing cultures were selected and incubated at higher product concentrations. An example of the performance of a selected culture is shown in Figure 2. Separations - Several membranes were considered. Testing was performed on a new product line produced by Sulzer Chemtech (Germany). These are tubular ceramic membranes with weak acid functionality (see Figure 3). The following results were observed: (1) The membranes were relatively fragile in a laboratory setting; (2) Thermally stable {at} 130 C in hot organic acids; (3) Acetic acid rejection > 99%; and (4) Moderate ammonia flux. The advantages of producing acetic acid by fermentation include its appropriateness for small-scale production, lower cost feedstocks, low energy membrane-based purification, and lower temperature and pressure requirements. Potential energy savings of using fermentation are estimated to be approximately 14 trillion Btu by 2020 from a reduction in natural gas use. Decreased transportation needs with regional plants will eliminate approximately 200 million gallons of diesel consumption, for combined savings of 45 trillion Btu. If the fermentation process captures new acetic acid production, savings could include an additional 5 trillion Btu from production and 7 trillion Btu from transportation energy.

  18. Mathematical models of continuous flow electrophoresis: Electrophoresis technology

    NASA Technical Reports Server (NTRS)

    Saville, Dudley A.

    1986-01-01

    Two aspects of continuous flow electrophoresis were studied: (1) the structure of the flow field in continuous flow devices; and (2) the electrokinetic properties of suspended particles relevant to electrophoretic separations. Mathematical models were developed to describe flow structure and stability, with particular emphasis on effects due to buoyancy. To describe the fractionation of an arbitrary particulate sample by continuous flow electrophoresis, a general mathematical model was constructed. In this model, chamber dimensions, field strength, buffer composition, and other design variables can be altered at will to study their effects on resolution and throughput. All these mathematical models were implemented on a digital computer and the codes are available for general use. Experimental and theoretical work with particulate samples probed how particle mobility is related to buffer composition. It was found that ions on the surface of small particles are mobile, contrary to the widely accepted view. This influences particle mobility and suspension conductivity. A novel technique was used to measure the mobility of particles in concentrated suspensions.

  19. Integrated multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S. (Ames, IA); Tan, Hongdong (Ames, IA)

    2002-05-14

    The present invention provides an integrated multiplexed capillary electrophoresis system for the analysis of sample analytes. The system integrates and automates multiple components, such as chromatographic columns and separation capillaries, and further provides a detector for the detection of analytes eluting from the separation capillaries. The system employs multiplexed freeze/thaw valves to manage fluid flow and sample movement. The system is computer controlled and is capable of processing samples through reaction, purification, denaturation, pre-concentration, injection, separation and detection in parallel fashion. Methods employing the system of the invention are also provided.

  20. 21 CFR 522.2476 - Trenbolone acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...milligrams (mg) trenbolone acetate (one implant consisting of 7 pellets, each pellet containing 20 mg trenbolone acetate) per implant dose. (B) 140 mg trenbolone acetate (one implant consisting of 8 pellets, each of 7...

  1. 21 CFR 184.1721 - Sodium acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...animal tissues. Sodium acetate may occur in either the anhydrous or trihydrated form. It is produced synthetically by the neutralization of acetic acid with sodium carbonate or by treating calcium acetate with sodium sulfate and sodium...

  2. 21 CFR 184.1721 - Sodium acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...animal tissues. Sodium acetate may occur in either the anhydrous or trihydrated form. It is produced synthetically by the neutralization of acetic acid with sodium carbonate or by treating calcium acetate with sodium sulfate and sodium...

  3. 21 CFR 184.1721 - Sodium acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...animal tissues. Sodium acetate may occur in either the anhydrous or trihydrated form. It is produced synthetically by the neutralization of acetic acid with sodium carbonate or by treating calcium acetate with sodium sulfate and sodium...

  4. Capillary electrophoresis for drug analysis

    NASA Astrophysics Data System (ADS)

    Lurie, Ira S.

    1999-02-01

    Capillary electrophoresis (CE) is a high resolution separation technique which is amenable to a wide variety of solutes, including compounds which are thermally degradable, non-volatile and highly polar, and is therefore well suited for drug analysis. Techniques which have been used in our laboratory include electrokinetic chromatography (ECC), free zone electrophoresis (CZE) and capillary electrochromatography (CEC). ECC, which uses a charged run buffer additive which migrates counter to osmotic flow, is excellent for many applications, including, drug screening and analyses of heroin, cocaine and methamphetamine samples. ECC approaches include the use of micelles and charged cyclodextrins, which allow for the separation of complex mixtures. Simultaneous separation of acidic, neutral and basic solutes and the resolution of optical isomers and positional isomers are possible. CZE has been used for the analysis of small ions (cations and anions) in heroin exhibits. For the ECC and CZE experiments performed in our laboratory, uncoated capillaries were used. In contrast, CEC uses capillaries packed with high performance liquid chromatography stationary phases, and offers both high peak capacities and unique selectivities. Applications include the analysis of cannabinoids and drug screening. Although CE suffers from limited concentration sensitivity, it is still applicable to trace analysis of drug samples, especially when using injection techniques such as stacking, or detection schemes such as laser induced fluorescence and extended pathlength UV.

  5. DNA Sequencing Using capillary Electrophoresis

    SciTech Connect

    Dr. Barry Karger

    2011-05-09

    The overall goal of this program was to develop capillary electrophoresis as the tool to be used to sequence for the first time the Human Genome. Our program was part of the Human Genome Project. In this work, we were highly successful and the replaceable polymer we developed, linear polyacrylamide, was used by the DOE sequencing lab in California to sequence a significant portion of the human genome using the MegaBase multiple capillary array electrophoresis instrument. In this final report, we summarize our efforts and success. We began our work by separating by capillary electrophoresis double strand oligonucleotides using cross-linked polyacrylamide gels in fused silica capillaries. This work showed the potential of the methodology. However, preparation of such cross-linked gel capillaries was difficult with poor reproducibility, and even more important, the columns were not very stable. We improved stability by using non-cross linked linear polyacrylamide. Here, the entangled linear chains could move when osmotic pressure (e.g. sample injection) was imposed on the polymer matrix. This relaxation of the polymer dissipated the stress in the column. Our next advance was to use significantly lower concentrations of the linear polyacrylamide that the polymer could be automatically blown out after each run and replaced with fresh linear polymer solution. In this way, a new column was available for each analytical run. Finally, while testing many linear polymers, we selected linear polyacrylamide as the best matrix as it was the most hydrophilic polymer available. Under our DOE program, we demonstrated initially the success of the linear polyacrylamide to separate double strand DNA. We note that the method is used even today to assay purity of double stranded DNA fragments. Our focus, of course, was on the separation of single stranded DNA for sequencing purposes. In one paper, we demonstrated the success of our approach in sequencing up to 500 bases. Other application papers of sequencing up to this level were also published in the mid 1990's. A major interest of the sequencing community has always been read length. The longer the sequence read per run the more efficient the process as well as the ability to read repeat sequences. We therefore devoted a great deal of time to studying the factors influencing read length in capillary electrophoresis, including polymer type and molecule weight, capillary column temperature, applied electric field, etc. In our initial optimization, we were able to demonstrate, for the first time, the sequencing of over 1000 bases with 90% accuracy. The run required 80 minutes for separation. Sequencing of 1000 bases per column was next demonstrated on a multiple capillary instrument. Our studies revealed that linear polyacrylamide produced the longest read lengths because the hydrophilic single strand DNA had minimal interaction with the very hydrophilic linear polyacrylamide. Any interaction of the DNA with the polymer would lead to broader peaks and lower read length. Another important parameter was the molecular weight of the linear chains. High molecular weight (> 1 MDA) was important to allow the long single strand DNA to reptate through the entangled polymer matrix. In an important paper, we showed an inverse emulsion method to prepare reproducibility linear polyacrylamide polymer with an average MWT of 9MDa. This approach was used in the polymer for sequencing the human genome. Another critical factor in the successful use of capillary electrophoresis for sequencing was the sample preparation method. In the Sanger sequencing reaction, high concentration of salts and dideoxynucleotide remained. Since the sample was introduced to the capillary column by electrokinetic injection, these salt ions would be favorably injected into the column over the sequencing fragments, thus reducing the signal for longer fragments and hence reading read length. In two papers, we examined the role of individual components from the sequencing reaction and then developed a protocol to reduce the deleterio

  6. Compensating for Electro-Osmosis in Electrophoresis

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1987-01-01

    Simple mechanical adjustment eliminates transverse velocity component. New apparatus for moving-wall electrophoresis increases degree of collimation of chemical species in sample stream. Electrophoresis chamber set at slight angle in horizontal plane to adjust angle between solution flow and wall motion. Component of velocity created cancels electro-osmotic effect.

  7. Getting the Most out of Electrophoresis Units

    ERIC Educational Resources Information Center

    Mulvihill, Charlotte

    2007-01-01

    At Oklahoma City Community College, they have developed gel electrophoresis activities that support active learning of many scientific concepts, including: pH, electrolysis, oxidation reduction, electrical currents, potentials, conductivity, molarity, gel electrophoresis, DNA and protein separation, and DNA fingerprinting. This article presents…

  8. Patch Clamp Detection in Capillary Electrophoresis

    E-print Network

    Articles Patch Clamp Detection in Capillary Electrophoresis Kent Jardemark Department of Anatomy a capillary electrophoresis-patch clamp (CE- PC) analysis of biomolecules that activate ligand-gated ion responses were calculated from currents recorded with patch clamp detection. This information

  9. Fluorescence detection for gel and capillary electrophoresis

    SciTech Connect

    Hogan, B.

    1992-07-21

    First, an indirect fluorescence detection system for the separation of proteins via gel electrophoresis. Quantities as low as 50 nanograms of bovine serum albumin and soybean trypsin inhibitor are separated and detected visually without the need for staining of the analytes. This is very similar to levels of protein commonly separated with gel electrophoresis.

  10. Two Electrophoresis Experiments for Freshmen in the Health Professions.

    ERIC Educational Resources Information Center

    Brabson, G. Dana; Waugh, David S.

    1986-01-01

    Describes procedures involved with paper electrophoresis separation of amino acids, gel electrophoresis separation of DNA, and design of an electrophoresis tank. Describes experiments using paper (amino acids) and gel (deoxyribonucleic acid fragments). Provides material lists, procedures, and discussion. (JM)

  11. Electrophoresis-mass spectrometry probe

    DOEpatents

    Andresen, B.D.; Fought, E.R.

    1987-11-10

    The invention involves a new technique for the separation of complex mixtures of chemicals, which utilizes a unique interface probe for conventional mass spectrometers which allows the electrophoretically separated compounds to be analyzed in real-time by a mass spectrometer. This new chemical analysis interface, which couples electrophoresis with mass spectrometry, allows complex mixtures to be analyzed very rapidly, with much greater specificity, and with greater sensitivity. The interface or probe provides a means whereby large and/or polar molecules in complex mixtures to be completely characterized. The preferred embodiment of the probe utilizes a double capillary tip which allows the probe tip to be continually wetted by the buffer, which provides for increased heat dissipation, and results in a continually operating interface which is more durable and electronically stable than the illustrated single capillary tip probe interface. 8 figs.

  12. Electrophoresis-mass spectrometry probe

    DOEpatents

    Andresen, Brian D. (Pleasanton, CA); Fought, Eric R. (Livermore, CA)

    1987-01-01

    The invention involves a new technique for the separation of complex mixtures of chemicals, which utilizes a unique interface probe for conventional mass spectrometers which allows the electrophoretically separated compounds to be analyzed in real-time by a mass spectrometer. This new chemical analysis interface, which couples electrophoresis with mass spectrometry, allows complex mixtures to be analyzed very rapidly, with much greater specificity, and with greater sensitivity. The interface or probe provides a means whereby large and/or polar molecules in complex mixtures to be completely characterized. The preferred embodiment of the probe utilizes a double capillary tip which allows the probe tip to be continually wetted by the buffer, which provides for increased heat dissipation, and results in a continually operating interface which is more durable and electronically stable than the illustrated single capillary tip probe interface.

  13. Comparison of non-electrophoresis grade with electrophoresis grade BIS in NIPAM polymer gel preparation

    PubMed Central

    Khodadadi, Roghayeh; Khajeali, Azim; Farajollahi, Ali Reza; Hajalioghli, Parisa; Raeisi, Noorallah

    2015-01-01

    Introduction:The main objective of this study was to investigate the possibility of replacing electrophoresis cross-linker with non-electrophoresis N, N?-methylenebisacrylamide (BIS) in N-isopropyl acrylamide (NIPAM) polymer gel and its possible effect on dose response. Methods: NIPAM polymer gel was prepared from non-electrophoresis grade BIS and the relaxation rate (R2) was measured by MR imaging after exposing the gel to gamma radiation from Co-60 source. To compare the response of this gel with the one that contains electrophoresis grade BIS, two sets of NIPAM gel were prepared using electrophoresis and non-electrophoresis BIS and irradiated to different gamma doses. Results: It was found that the dose–response of NIPAM gel made from the non-electrophoresis grade BIS is coincident with that of electrophoresis grade BIS. Conclusion:Taken all, it can be concluded that the non-electrophoresis grade BIS not only is a suitable alternative for the electrophoresis grade BIS but also reduces the cost of gel due to its lower price. PMID:26457250

  14. The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis

    PubMed Central

    Loening, U. E.

    1967-01-01

    1. Gels were prepared with recrystallized acrylamide and bisacrylamide. Electrophoresis was in tris–sodium acetate–EDTA buffer for 0·5 to 3hr. Gels were scanned at 280 or 265m?. Techniques are described for slicing and radioactive counting. 2. The mobility of RNA was inversely related to the sedimentation coefficient and varied with gel concentration. Electrophoresis in 2·2–2·6% gels gives a fractionation similar to density-gradient centrifugation. It shows the two ribosomal RNA components, the 45s precursor, transfer RNA and minor components. In 5% and 7·5% gels, 4s and 5s RNA separated and ribosomal RNA was excluded. 3. The resolution is greater and more detailed than by centrifugation, and many samples can be analysed simultaneously and rapidly. PMID:5339944

  15. Micelles Protect and Concentrate Activated Acetic Acid

    NASA Astrophysics Data System (ADS)

    Todd, Zoe; House, C.

    2014-01-01

    As more and more exoplanets are discovered and the habitability of such planets is considered, one can turn to searching for the origin of life on Earth in order to better understand what makes a habitable planet. Activated acetic acid, or methyl thioacetate, has been proposed to be central to the origin of life on Earth, and also as an important energy currency molecule in early cellular evolution. We have investigated the hydrolysis of methyl thioacetate under various conditions. Its uncatalyzed rate of hydrolysis is about three orders of magnitude faster (K = 0.00663 s^-1; 100°C, pH 7.5, concentration = 0.33mM) than published rates for its catalyzed production making it unlikely to accumulate under prebiotic conditions. However, we also observed that methyl thioacetate was protected from hydrolysis when inside its own hydrophobic droplets. We found that methyl thioacetate protection from hydrolysis was also possible in droplets of hexane and in the membranes of nonanoic acid micelles. Thus, the hydrophobic regions of prebiotic micelles and early cell membranes could have offered a refuge for this energetic molecule increasing its lifetime in close proximity to the reactions for which it would be needed. Methyl thioacetate could thus be important for the origin of life on Earth and perhaps for better understanding the potential habitability of other planets.

  16. Nonlinear waves in capillary electrophoresis

    PubMed Central

    Ghosal, Sandip; Chen, Zhen

    2011-01-01

    Electrophoretic separation of a mixture of chemical species is a fundamental technique of great usefulness in biology, health care and forensics. In capillary electrophoresis the sample migrates in a microcapillary in the presence of a background electrolyte. When the ionic concentration of the sample is sufficiently high, the signal is known to exhibit features reminiscent of nonlinear waves including sharp concentration ‘shocks’. In this paper we consider a simplified model consisting of a single sample ion and a background electrolyte consisting of a single co-ion and a counterion in the absence of any processes that might change the ionization states of the constituents. If the ionic diffusivities are assumed to be the same for all constituents the concentration of sample ion is shown to obey a one dimensional advection diffusion equation with a concentration dependent advection velocity. If the analyte concentration is sufficiently low in a suitable non-dimensional sense, Burgers’ equation is recovered, and thus, the time dependent problem is exactly solvable with arbitrary initial conditions. In the case of small diffusivity either a leading edge or trailing edge shock is formed depending on the electrophoretic mobility of the sample ion relative to the background ions. Analytical formulas are presented for the shape, width and migration velocity of the sample peak and it is shown that axial dispersion at long times may be characterized by an effective diffusivity that is exactly calculated. These results are consistent with known observations from physical and numerical simulation experiments. PMID:20238181

  17. Mesoscale modelling of polyelectrolyte electrophoresis.

    PubMed

    Grass, Kai; Holm, Christian

    2010-01-01

    The electrophoretic behaviour of flexible polyelectrolyte chains ranging from single monomers up to long fragments of a hundred repeat units is studied by a mesoscopic simulation approach. Abstracting from the atomistic details of the polyelectrolyte and the fluid, a coarse-grained molecular dynamics model connected to a mesoscopic fluid described by the Lattice-Boltzmann approach is used to investigate free-solution electrophoresis. Our study demonstrates the importance of hydrodynamic interactions for the electrophoretic motion of polyelectrolytes and quantifies the influence of surrounding ions. The length-dependence of the electrophoretic mobility can be understood by evaluating the scaling behavior of the effective charge and the effective friction. The perfect agreement of our results with experimental measurements shows that all chemical details and fluid structure can be safely neglected, and a suitable coarse-grained approach can yield an accurate description of the physics of the problem, provided that electrostatic and hydrodynamic interactions between all entities in the system, i.e., the polyelectrolyte, dissociated counterions, additional salt and the solvent, are properly accounted for. Our model is able to bridge the single molecule regime of a few nm up to macromolecules with contour lengths of more than 100 nm, a length scale that is currently not accessible to atomistic simulations. PMID:20158023

  18. Mesoscale modelling of polyelectrolyte electrophoresis

    E-print Network

    Kai Grass; Christian Holm

    2009-02-11

    The electrophoretic behaviour of flexible polyelectrolyte chains ranging from single monomers up to long fragments of hundred repeat units is studied by a mesoscopic simulation approach. Abstracting from the atomistic details of the polyelectrolyte and the fluid, a coarse-grained molecular dynamics model connected to a mesoscopic fluid described by the Lattice Boltzmann approach is used to investigate free-solution electrophoresis. Our study demonstrates the importance of hydrodynamic interactions for the electrophoretic motion of polyelectrolytes and quantifies the influence of surrounding ions. The length-dependence of the electrophoretic mobility can be understood by evaluating the scaling behavior of the effective charge and the effective friction. The perfect agreement of our results with experimental measurements shows that all chemical details and fluid structure can be safely neglected, and a suitable coarse-grained approach can yield an accurate description of the physics of the problem, provided that electrostatic and hydrodynamic interactions between all entities in the system, i.e., the polyelectrolyte, dissociated counterions, additional salt and the solvent, are properly accounted for. Our model is able to bridge the single molecule regime of a few nm up to macromolecules with contour lengths of more than 100 nm, a length scale that is currently not accessible to atomistic simulations.

  19. Vibrational spectroscopy and electrophoresis as a "golden means" in monitoring of polysaccharides in medical plant and gels.

    PubMed

    Pielesz, A

    2012-07-01

    In recent years, some bioactive polysaccharides isolated from natural sources have attracted much attention in the field of biochemistry and pharmacology. Of them, polysaccharides or their glycoconjugates were shown to exhibit multiple biological activities including anticarcinogenic, anticoagulant, immunostimulating, antioxidant, etc. Pharmacotherapy using plant-derived substances can be currently regarded as a very promising future alternative to conventional therapy. The advanced biotechnologies available today enable chemical investigation of well-defined bioactive plant components as sources of novel drugs. The need for safer drugs without side effects has led to the use of natural ingredients with proven safety. Special interest is focused on plant polysaccharides. This article attempts to review the current structural and conformational characterization of some importantly bioactive monosaccharides isolated from following plant cell-wall: Symphytum officinale (comfrey), Thymus pulegioides (thyme), Trigonella foenum-graecum L. (fenugreek), Tussilago farfara L. (coltsfoot), Hyssopus officinalis (hyssop), Althaea officinalis L. (marshmallow) and Equisetum arvense L. (horsetail). The chemical structures of monosaccharides were analysed using FTIR and Raman spectroscopies as well as cellulose acetate membrane electrophoresis (CAE). The dried plant samples were gently hydrolysed with sulphuric acid. The presence of glucuronic acid, galacturonic acid, alginic acid, glucose, mannose and xylose in the hydrolysates of reference substances and non-defatted plant films was proved. The possibility of a taxonomic classification of plant cell walls based on infrared and Raman spectroscopies and the use of spectral fingerprinting for authentication and detection of adulteration of products rich in cell-wall materials are discussed. Individual bands were selected to monitor the sugar content in medical plant cell walls and to confirm the identity of the analysed plants. PMID:22465769

  20. Vibrational spectroscopy and electrophoresis as a "golden means" in monitoring of polysaccharides in medical plant and gels

    NASA Astrophysics Data System (ADS)

    Pielesz, A.

    In recent years, some bioactive polysaccharides isolated from natural sources have attracted much attention in the field of biochemistry and pharmacology. Of them, polysaccharides or their glycoconjugates were shown to exhibit multiple biological activities including anticarcinogenic, anticoagulant, immunostimulating, antioxidant, etc. Pharmacotherapy using plant-derived substances can be currently regarded as a very promising future alternative to conventional therapy. The advanced biotechnologies available today enable chemical investigation of well-defined bioactive plant components as sources of novel drugs. The need for safer drugs without side effects has led to the use of natural ingredients with proven safety. Special interest is focused on plant polysaccharides. This article attempts to review the current structural and conformational characterization of some importantly bioactive monosaccharides isolated from following plant cell-wall: Symphytum officinale (comfrey), Thymus pulegioides (thyme), Trigonella foenum-graecum L. (fenugreek), Tussilago farfara L. (coltsfoot), Hyssopus officinalis (hyssop), Althaea officinalis L. (marshmallow) and Equisetum arvense L. (horsetail). The chemical structures of monosaccharides were analysed using FTIR and Raman spectroscopies as well as cellulose acetate membrane electrophoresis (CAE). The dried plant samples were gently hydrolysed with sulphuric acid. The presence of glucuronic acid, galacturonic acid, alginic acid, glucose, mannose and xylose in the hydrolysates of reference substances and non-defatted plant films was proved. The possibility of a taxonomic classification of plant cell walls based on infrared and Raman spectroscopies and the use of spectral fingerprinting for authentication and detection of adulteration of products rich in cell-wall materials are discussed. Individual bands were selected to monitor the sugar content in medical plant cell walls and to confirm the identity of the analysed plants.

  1. Capillary electrophoresis electrospray ionization mass spectrometry interface

    DOEpatents

    Smith, Richard D. (Richland, WA); Severs, Joanne C. (Hayward, CA)

    1999-01-01

    The present invention is an interface between a capillary electrophoresis separation capillary end and an electrospray ionization mass spectrometry emitter capillary end, for transporting an anolyte sample from a capillary electrophoresis separation capillary to a electrospray ionization mass spectrometry emitter capillary. The interface of the present invention has: (a) a charge transfer fitting enclosing both of the capillary electrophoresis capillary end and the electrospray ionization mass spectrometry emitter capillary end; (b) a reservoir containing an electrolyte surrounding the charge transfer fitting; and (c) an electrode immersed into the electrolyte, the electrode closing a capillary electrophoresis circuit and providing charge transfer across the charge transfer fitting while avoiding substantial bulk fluid transfer across the charge transfer fitting. Advantages of the present invention have been demonstrated as effective in providing high sensitivity and efficient analyses.

  2. Study of disposable microdevices for DNA electrophoresis

    E-print Network

    Timp, Winston (Winston G.)

    2005-01-01

    A study was undertaken to determine if a microfluidic chip, made of economical plastic materials, is feasible. The chip was designed to perform gel electrophoresis, specifically of DNA fragments for either sequencing or ...

  3. High-performance capillary electrophoresis of histones

    SciTech Connect

    Gurley, L.R.; London, J.E.; Valdez, J.G.

    1991-01-01

    A high performance capillary electrophoresis (HPCE) system has been developed for the fractionation of histones. This system involves electroinjection of the sample and electrophoresis in a 0.1M phosphate buffer at pH 2.5 in a 50 {mu}m {times} 35 cm coated capillary. Electrophoresis was accomplished in 9 minutes separating a whole histone preparation into its components in the following order of decreasing mobility; (MHP) H3, H1 (major variant), H1 (minor variant), (LHP) H3, (MHP) H2A (major variant), (LHP) H2A, H4, H2B, (MHP) H2A (minor variant) where MHP is the more hydrophobic component and LHP is the less hydrophobic component. This order of separation is very different from that found in acid-urea polyacrylamide gel electrophoresis and in reversed-phase HPLC and, thus, brings the histone biochemist a new dimension for the qualitative analysis of histone samples. 27 refs., 8 figs.

  4. Induced-charge electrophoresis near a wall

    E-print Network

    Kilic, Mustafa Sabri

    Induced-charge electrophoresis (ICEP) has mostly been analyzed for asymmetric particles in an infinite fluid, but channel walls in real systems further break symmetry and lead to dielectrophoresis (DEP) in local field ...

  5. Preparation of Electrically Conductive Polymeric Membranes

    NASA Astrophysics Data System (ADS)

    Encinas, J. C.; Castillo-Ortega, M. M.; Rodríguez, F.; Castaño, V. M.

    2015-10-01

    Cellulose acetate porous membranes, coated with polyaniline, were chemically modified with polyelectrolytes to produce films of varying and controlled porosity and electrical conductivity. The highest electrical conductivity was obtained in membranes prepared with poly(styrene sulfonate) with large pore sizes. The electrical properties as well as scanning electron microscopy (SEM) images are discussed.

  6. Affinity Electrophoresis Using Ligands Attached To Polymers

    NASA Technical Reports Server (NTRS)

    Van Alstine, James M.; Snyder, Robert S.; Harris, J. M.; Brooks, D. E.

    1990-01-01

    In new technique, reduction of electrophoretic mobilities by addition of polyethylene glycol to ligands increases electrophoretic separabilities. In immuno-affinity electrophoresis, modification of ligands extends specificity of electrophoretic separation to particles having surface electric-charge structures otherwise making them electrophoretically inseparable. Modification of antibodies by polyethylene glycol greatly reduces ability to aggregate while enhancing ability to affect electrophoretic mobilities of cells. In hydrophobic-affinity electrophoresis, addition of polyethylene glycol reduces tendency toward aggregation of cells or macromolecules.

  7. Free-Flow Open-Chamber Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Sammons, David W.

    1994-01-01

    Free-flow open-chamber electrophoresis variant of free-flow electrophoresis performed in chamber with open ends and in which velocity of electro-osmotic flow adjusted equal to and opposite mean electrophoretic velocity of sample. Particles having electrophoretic mobilities greater than mean mobility of sample particles move toward cathode, those with mobilities less move toward anode. Technique applied to separation of components of mixtures of biologically important substances. Sensitivity enhanced by use of tapered chamber.

  8. Reductive opening of carbohydrate phenylsulfonylethylidene (PSE) acetals.

    PubMed

    Chéry, Florence; Cabianca, Elena; Tatibouët, Arnaud; De Lucchi, Ottorino; Lindhorst, Thisbe K; Rollin, Patrick

    2015-11-19

    The phenylsulfonylethylidene (PSE) acetal is a relatively new protecting group in carbohydrate chemistry. However, carbohydrate-derived phenylsulfonylethylidene (PSE) acetals show a different behavior in reductive desulfonylation than simple symmetrical acetals. Here we have investigated various SET-type reaction conditions in order to open PSE acetals regioselectively and to produce chiral ?-hydroxyethenyl ethers. Whereas sodium amalgam leads to a mixture of regioisomeric vinyl ethers besides the ethylidene acetal, samarium iodide is suited for regioselective ring opening. This is shown with seven different carbohydrate PSE acetals, both of the 1,3-dioxane and the 1,3-dioxolane type. PMID:26469209

  9. Fragrance material review on 2,4-dimethylbenzyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 2,4-dimethylbenzyl acetate when used as a fragrance ingredient is presented. 2,4-Dimethylbenzyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, iso-butyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 2,4-dimethylbenzyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, and skin sensitization data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414641

  10. Fragrance material review on p-anisyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of p-anisyl acetate when used as a fragrance ingredient is presented. p-Anisyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for p-anisyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, mucous membrane (eye) irritation, skin sensitization, and genotoxicity data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22417777

  11. 21 CFR 522.2476 - Trenbolone acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL...milligrams (mg) trenbolone acetate (one implant consisting of 7 pellets, each pellet containing 20 mg trenbolone acetate) per implant dose. (B) 140 mg trenbolone...

  12. Ozone decomposition in aqueous acetate solutions

    SciTech Connect

    Sehested, K.; Holcman, J.; Bjergbakke, E.; Hart, E.J.

    1987-01-01

    The acetate radical ion reacts with ozone with a rate constant of k = (1.5 +/- 0.5) x 10Z dmT mol s . The products from this reaction are CO2, HCHO, and O2 . By subsequent reaction of the peroxy radical with ozone the acetate radical ion is regenerated through the OH radical. A chain decomposition of ozone takes place. It terminates when the acetate radical ion reacts with oxygen forming the unreactive peroxy acetate radical. The chain is rather short as oxygen is developed, as a result of the ozone consumption. The inhibiting effect of acetate on the ozone decay is rationalized by OH scavenging by acetate and successive reaction of the acetate radical ion with oxygen. Some products from the bimolecular disappearance of the peroxy acetate radicals, however, react further with ozone, reducing the effectiveness of the stabilization.

  13. Effect of membrane polymeric materials on relationship between surface pore size and membrane fouling in membrane bioreactors

    NASA Astrophysics Data System (ADS)

    Miyoshi, Taro; Yuasa, Kotaku; Ishigami, Toru; Rajabzadeh, Saeid; Kamio, Eiji; Ohmukai, Yoshikage; Saeki, Daisuke; Ni, Jinren; Matsuyama, Hideto

    2015-03-01

    We investigated the effect of different membrane polymeric materials on the relationship between membrane pore size and development of membrane fouling in a membrane bioreactor (MBR). Membranes with different pore sizes were prepared using three different polymeric materials, cellulose acetate butyrate (CAB), polyvinyl butyral (PVB), and polyvinylidene fluoride (PVDF), and the development of membrane fouling in each membrane was evaluated by batch filtration tests using a mixed liquor suspension obtained from a laboratory-scale MBR. The results revealed that the optimal membrane pore size to mitigate membrane fouling differed depending on membrane polymeric material. For PVDF membranes, the degree of membrane fouling decreased as membrane pore size increased. In contrast, CAB membranes with smaller pores had less fouling propensity than those with larger ones. Such difference can be attributed to the difference in major membrane foulants in each membrane; in PVDF, they were small colloids or dissolved organics in which proteins are abundant, and in CAB, microbial flocs. The results obtained in this study strongly suggested that optimum operating conditions of MBRs differ depending on the characteristics of the used membrane.

  14. 21 CFR 582.1721 - Sodium acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium acetate. 582.1721 Section 582.1721 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1721 Sodium acetate. (a) Product. Sodium acetate. (b) Conditions of use. This substance is...

  15. 21 CFR 582.1721 - Sodium acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium acetate. 582.1721 Section 582.1721 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1721 Sodium acetate. (a) Product. Sodium acetate. (b) Conditions of use. This substance is...

  16. 21 CFR 582.1721 - Sodium acetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Sodium acetate. 582.1721 Section 582.1721 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1721 Sodium acetate. (a) Product. Sodium acetate. (b) Conditions of use. This substance is...

  17. 21 CFR 582.1721 - Sodium acetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium acetate. 582.1721 Section 582.1721 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1721 Sodium acetate. (a) Product. Sodium acetate. (b) Conditions of use. This substance is...

  18. 21 CFR 582.1721 - Sodium acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium acetate. 582.1721 Section 582.1721 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1721 Sodium acetate. (a) Product. Sodium acetate. (b) Conditions of use. This substance is...

  19. 21 CFR 582.1005 - Acetic acid.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Acetic acid. 582.1005 Section 582.1005 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1005 Acetic acid. (a) Product. Acetic acid. (b) Conditions of use. This substance is...

  20. 21 CFR 582.1005 - Acetic acid.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Acetic acid. 582.1005 Section 582.1005 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1005 Acetic acid. (a) Product. Acetic acid. (b) Conditions of use. This substance is...

  1. 21 CFR 582.1005 - Acetic acid.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Acetic acid. 582.1005 Section 582.1005 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1005 Acetic acid. (a) Product. Acetic acid. (b) Conditions of use. This substance is...

  2. 21 CFR 582.1005 - Acetic acid.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Acetic acid. 582.1005 Section 582.1005 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1005 Acetic acid. (a) Product. Acetic acid. (b) Conditions of use. This substance is...

  3. 21 CFR 582.1005 - Acetic acid.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Acetic acid. 582.1005 Section 582.1005 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS....1005 Acetic acid. (a) Product. Acetic acid. (b) Conditions of use. This substance is...

  4. 21 CFR 184.1185 - Calcium acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...known as acetate of lime or vinegar salts, is the calcium salt of acetic acid. It may be produced by the calcium hydroxide neutralization of acetic acid. (b) The ingredient meets the specifications of the Food Chemicals Codex, 3d Ed....

  5. 21 CFR 184.1185 - Calcium acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...known as acetate of lime or vinegar salts, is the calcium salt of acetic acid. It may be produced by the calcium hydroxide neutralization of acetic acid. (b) The ingredient meets the specifications of the Food Chemicals Codex, 3d Ed....

  6. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a...additive lead acetate may be safely used in cosmetics intended for coloring hair on the scalp...The amount of the lead acetate in the cosmetic shall be such that the lead...

  7. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a...additive lead acetate may be safely used in cosmetics intended for coloring hair on the scalp...The amount of the lead acetate in the cosmetic shall be such that the lead...

  8. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a...additive lead acetate may be safely used in cosmetics intended for coloring hair on the scalp...The amount of the lead acetate in the cosmetic shall be such that the lead...

  9. 21 CFR 522.2476 - Trenbolone acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Trenbolone acetate. 522.2476 Section 522.2476 Food... Trenbolone acetate. (a) Sponsors. See sponsors in § 510.600(c) of this chapter for use as in paragraph (d) of... days. (A) 140 milligrams (mg) trenbolone acetate (one implant consisting of 7 pellets, each...

  10. 21 CFR 522.2476 - Trenbolone acetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Trenbolone acetate. 522.2476 Section 522.2476 Food... Trenbolone acetate. (a) Sponsors. See sponsors in § 510.600(c) of this chapter for use as in paragraph (d) of... days. (A) 140 milligrams (mg) trenbolone acetate (one implant consisting of 7 pellets, each...

  11. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

    Studies of a zwitterionic buffer formulated for cell electrophoresis were done using the McDonnell-Douglas Continuous Flow Electrophoresis System. Standard buffers were analyzed for their stability in the electrical field and the results showed that both buffers tested were inherently unstable. Further, titration studies showed that the standards buffers buffered poorly at the pH employed for electrophoresis. The zwitterionic buffer buffered well at its nominal pH and was shown to be stable in the electrical field. Comparative studies of the buffer with standard cell separation buffers using formalin fixed rabbit and goose red blood cells showed that the zwitterionic buffer gave better resolution of the fixed cells. Studies with viable hybridoma cells showed that buffer Q supported cell viability equal to Hank's Balanced Salt Solution and that hybridoma cells in different stages of the growth cycle demonstrated reproducible differences in electrophoretic mobility.

  12. Undergraduate physics laboratory: Electrophoresis in chromatography paper

    NASA Astrophysics Data System (ADS)

    Hyde, Alexander; Batishchev, Oleg

    2015-12-01

    An experiment studying the physical principles of electrophoresis in liquids was developed for an undergraduate laboratory. We have improved upon the standard agarose gel electrophoresis experimental regime with a straightforward and cost-effective procedure, in which drops of widely available black food coloring were separated by electric field into their dye components on strips of chromatography paper soaked in a baking soda/water solution. Terminal velocities of seven student-safe dyes were measured as a function of the electric potential applied along the strips. The molecular mobility was introduced and calculated by analyzing data for a single dye. Sources of systematic and random errors were investigated.

  13. Acetic acid induces pH-independent cellular energy depletion in Salmonella enterica.

    PubMed

    Tan, Sin Mei; Lee, Sui Mae; Dykes, Gary A

    2015-03-01

    Weak organic acids are widely used as preservatives and disinfectants in the food industry. Despite their widespread use, the antimicrobial mode of action of organic acids is still not fully understood. This study investigated the effect of acetic acid on the cell membranes and cellular energy generation of four Salmonella strains. Using a nucleic acid/protein assay, it was established that acetic acid did not cause leakage of intracellular components from the strains. A scanning electron microscopy study further confirmed that membrane disruption was not the antimicrobial mode of action of acetic acid. Some elongated Salmonella cells observed in the micrographs indicated a possibility that acetic acid may inhibit DNA synthesis in the bacterial cells. Using an ATP assay, it was found that at a neutral pH, acetic acid caused cellular energy depletion with an ADP/ATP ratio in the range between 0.48 and 2.63 (p<0.05) that was apparent for the four Salmonella strains. We suggest that this effect was probably due solely to the action of undissociated acid molecules. The antimicrobial effect of acetic acid was better under acidic conditions (ADP/ATP ratio of 5.56 ± 1.27; p<0.05), where the role of both pH and undissociated acid molecules can act together. We concluded that the inhibitory effect of acetic acid is not solely attributable to acidic pH but also to undissociated acid molecules. This finding has implication for the use of acetic acid as an antimicrobial against Salmonella on food products, such as chicken meat, which can buffer its pH. PMID:25562466

  14. Using Gel Electrophoresis To Illustrate Protein Diversity and Isoelectric Point.

    ERIC Educational Resources Information Center

    Browning, Mark; Vanable, Joseph

    2002-01-01

    Demonstrates the differences in protein structures by focusing on isoelectric point with an experiment that is observable under certain pH levels in gel electrophoresis. Explains the electrophoresis procedure and reports results of the experiments. (YDS)

  15. DNA DAMAGE QUANTITATION BY ALKALINE GEL ELECTROPHORESIS.

    SciTech Connect

    SUTHERLAND,B.M.; BENNETT,P.V.; SUTHERLAND, J.C.

    2004-03-24

    Physical and chemical agents in the environment, those used in clinical applications, or encountered during recreational exposures to sunlight, induce damages in DNA. Understanding the biological impact of these agents requires quantitation of the levels of such damages in laboratory test systems as well as in field or clinical samples. Alkaline gel electrophoresis provides a sensitive (down to {approx} a few lesions/5Mb), rapid method of direct quantitation of a wide variety of DNA damages in nanogram quantities of non-radioactive DNAs from laboratory, field, or clinical specimens, including higher plants and animals. This method stems from velocity sedimentation studies of DNA populations, and from the simple methods of agarose gel electrophoresis. Our laboratories have developed quantitative agarose gel methods, analytical descriptions of DNA migration during electrophoresis on agarose gels (1-6), and electronic imaging for accurate determinations of DNA mass (7-9). Although all these components improve sensitivity and throughput of large numbers of samples (7,8,10), a simple version using only standard molecular biology equipment allows routine analysis of DNA damages at moderate frequencies. We present here a description of the methods, as well as a brief description of the underlying principles, required for a simplified approach to quantitation of DNA damages by alkaline gel electrophoresis.

  16. Temperature Effects on Electrophoresis Supporting Information

    E-print Network

    Santiago, Juan G.

    S-1 Temperature Effects on Electrophoresis Supporting Information Anita Rogacs, Juan G. Santiago contains the following supplementary figures, tables, and information further describing our temperature of ionization at room temperature, 25°C. · Predictions for anionic (and cationic) mobility for the solutions

  17. Increasing Sensitivity In Continuous-Flow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Sammons, David W.

    1994-01-01

    Sensitivity of continuous-flow electrophoresis (CFE) chamber increased by introducing lateral gradients in concentration of buffer solution and thickness of chamber. Such gradients, with resulting enhanced separation, achieved in CFE chamber with wedge-shaped cross section and collateral flow. Enables improved separations of homogeneous components of mixtures of variety of biologically important substances.

  18. Role of gravity in preparative electrophoresis

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1975-01-01

    The fundamental formulas of electrophoresis are derived microscopically and applied to the problem of isotachophoresis. A simple physical model of the isotachophoresis front is proposed. The front motion and structure are studied in the simplified case without convection, diffusion and non-electric external forces.

  19. A Simple Vertical Slab Gel Electrophoresis Apparatus.

    ERIC Educational Resources Information Center

    Carter, J. B.; And Others

    1983-01-01

    Describes an inexpensive, easily constructed, and safe vertical slab gel kit used routinely for sodium dodecyl sulphate-polyacrylamide gel electrophoresis research and student experiments. Five kits are run from a single transformer. Because toxic solutions are used, students are given plastic gloves and closely supervised during laboratory…

  20. Capillary Electrophoresis for the Analysis of Biopolymers

    E-print Network

    Krylov, Sergey

    Postreplication Modification 114R Proteins and Peptides 114R Separation 114R Detection 118R Posttranslational: nucleic acids, proteins, lipids, and carbohydrates. While this review formally covers 1998 and 1999, we-scale capillary array electrophoresis instruments have been marketed by PE Biosystems (the model 3700 Genetic

  1. Determination of egg white lysozyme by on-line coupled capillary isotachophoresis with capillary zone electrophoresis.

    PubMed

    Kvasnicka, Frantisek

    2003-03-01

    An on-line coupled capillary isotachophoresis - capillary zone electrophoresis method for the determination of lysozyme in selected food products is described. The optimized electrolyte system consisted of 10 mM NH(4)OH + 20 mM acetic acid (leading electrolyte), 5 mM epsilon -aminocaproic acid +5 mM acetic acid (terminating electrolyte), and 20 mM epsilon -aminocaproic acid +5 mM acetic acid +0.1% m/v hydroxypropylmethylcellulose (background electrolyte). A clear separation of lysozyme from other components of acidic sample extract was achieved within 15 min. Method characteristics, i.e., linearity (0-50 micrograms/mL), accuracy (recovery 96+/-5%), intra-assay (3.8%), quantification limit (1 microgram/ml), and detection limit (0.25 microgram/mL) were determined. Low laboriousness, sufficient sensitivity and low running costs are important attributes of this method. The developed method is suitable for the quantification of the egg content in egg pasta. PMID:12627448

  2. Enzymatic production of glycerol acetate from glycerol.

    PubMed

    Oh, Seokhyeon; Park, Chulhwan

    2015-02-01

    In this study, we report the enzymatic production of glycerol acetate from glycerol and methyl acetate. Lipases are essential for the catalysis of this reaction. To find the optimum conditions for glycerol acetate production, sequential experiments were designed. Type of lipase, lipase concentration, molar ratio of reactants, reaction temperature and solvents were investigated for the optimum conversion of glycerol to glycerol acetate. As the result of lipase screening, Novozym 435 (Immobilized Candida antarctica lipase B) was turned out to be the optimal lipase for the reaction. Under the optimal conditions (2.5 g/L of Novozym 435, 1:40 molar ratio of glycerol to methyl acetate, 40 °C and tert-butanol as the solvent), glycerol acetate production was achieved in 95.00% conversion. PMID:25640720

  3. Detection of glycoproteins in the Acanthamoeba plasma membrane

    SciTech Connect

    Paatero, G.I.L. ); Gahmberg, C.G. )

    1988-11-01

    In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by {sup 125}I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB{sup 3}H{sub 4} and galactose oxidase/NaB{sup 3}H{sub 4} labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M{sub r} of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presence of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with ({sup 35}S)methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis.

  4. Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

    DOEpatents

    Zhang, Jian-Shi (Shanghai, CN); Giometti, Carol S. (Glenview, IL); Tollaksen, Sandra L. (Montgomery, IL)

    1989-01-01

    After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a DC power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, so that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. A high percentage extraction of proteins is achieved. The extracted proteins can be removed and subjected to partial digestion by trypsin or the like, followed by two-dimensional electrophoresis, resulting in a gel slab having a pattern of peptide gel spots which can be cored out and subjected to electrophoretic extraction to extract individual peptides.

  5. Numerical simulation of electrophoresis separation processes

    NASA Technical Reports Server (NTRS)

    Ganjoo, D. K.; Tezduyar, T. E.

    1986-01-01

    A new Petrov-Galerkin finite element formulation has been proposed for transient convection-diffusion problems. Most Petrov-Galerkin formulations take into account the spatial discretization, and the weighting functions so developed give satisfactory solutions for steady state problems. Though these schemes can be used for transient problems, there is scope for improvement. The schemes proposed here, which consider temporal as well as spatial discretization, provide improved solutions. Electrophoresis, which involves the motion of charged entities under the influence of an applied electric field, is governed by equations similiar to those encountered in fluid flow problems, i.e., transient convection-diffusion equations. Test problems are solved in electrophoresis and fluid flow. The results obtained are satisfactory. It is also expected that these schemes, suitably adapted, will improve the numerical solutions of the compressible Euler and the Navier-Stokes equations.

  6. A new approach to electrophoresis in space

    NASA Technical Reports Server (NTRS)

    Snyder, Robert S.; Rhodes, Percy H.

    1990-01-01

    Previous electrophoresis experiments performed in space are reviewed. There is sufficient data available from the results of these experiments to show that they were designed with incomplete knowledge of the fluid dynamics of the process including electrohydrodynamics. Redesigning laboratory chambers and operating procedures developed on Earth for space without understanding both the advantages and disadvantages of the microgravity environment has yielded poor separations of both cells and proteins. However, electrophoreris is still an important separation tool in the laboratory and thermal convection does limit its performance. Thus, there is a justification for electrophoresis but the emphasis of future space experiments must be directed toward basic research with model experiments to understand the microgravity environment and fluid analysis to test the basic principles of the process.

  7. Mathematical Models of Continuous Flow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.; Snyder, R. S.

    1985-01-01

    Development of high resolution continuous flow electrophoresis devices ultimately requires comprehensive understanding of the ways various phenomena and processes facilitate or hinder separation. A comprehensive model of the actual three dimensional flow, temperature and electric fields was developed to provide guidance in the design of electrophoresis chambers for specific tasks and means of interpreting test data on a given chamber. Part of the process of model development includes experimental and theoretical studies of hydrodynamic stability. This is necessary to understand the origin of mixing flows observed with wide gap gravitational effects. To insure that the model accurately reflects the flow field and particle motion requires extensive experimental work. Another part of the investigation is concerned with the behavior of concentrated sample suspensions with regard to sample stream stability particle-particle interactions which might affect separation in an electric field, especially at high field strengths. Mathematical models will be developed and tested to establish the roles of the various interactions.

  8. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, E.S.; Taylor, J.A.

    1994-06-28

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figures.

  9. Capillary zone electrophoresis-mass spectrometer interface

    DOEpatents

    D`Silva, A.

    1996-08-06

    A device for providing equal electrical potential between two loci unconnected by solid or liquid electrical conductors is provided. The device comprises a first electrical conducting terminal, a second electrical conducting terminal connected to the first terminal by a rigid dielectric structure, and an electrically conducting gas contacting the first and second terminals. This device is particularly suitable for application in the electrospray ionization interface between a capillary zone electrophoresis apparatus and a mass spectrometer. 1 fig.

  10. Capillary zone electrophoresis-mass spectrometer interface

    DOEpatents

    D'Silva, Arthur (Ames, IA)

    1996-08-06

    A device for providing equal electrical potential between two loci unconnected by solid or liquid electrical conducts is provided. The device comprises a first electrical conducting terminal, a second electrical conducting terminal connected to the first terminal by a rigid dielectric structure, and an electrically conducting gas contacting the first and second terminals. This device is particularly suitable for application in the electrospray ionization interface between a capillary zone electrophoresis apparatus and a mass spectrometer.

  11. Portable electrophoresis apparatus using minimum electrolyte

    NASA Technical Reports Server (NTRS)

    Stevens, M. R.; Vickers, J. M. (inventors)

    1976-01-01

    An electrophoresis unit for use in conducting electrophoretic analysis of specimens is described. The unit includes a sealable container in which a substrate mounted specimen is suspended in an electrolytic vapor. A heating unit is employed to heat a supply of electrolyte to produce the vapor. The substrate is suspended within the container by being attached between a pair of clips which also serve as electrodes to which a direct current power source may be connected.

  12. Method and apparatus for continuous electrophoresis

    DOEpatents

    Watson, Jack S. (Knoxville, TN)

    1992-01-01

    A method and apparatus for conducting continuous separation of substances by electrophoresis are disclosed. The process involves electrophoretic separation combined with couette flow in a thin volume defined by opposing surfaces. By alternating the polarity of the applied potential and producing reciprocating short rotations of at least one of the surfaces relative to the other, small increments of separation accumulate to cause substantial, useful segregation of electrophoretically separable components in a continuous flow system.

  13. Commander prepares glass columns for electrophoresis experiment

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Commander Jack Lousma prepares on of the glass columns for the electrophoresis test in the middeck area of the Columbia. The experiment, deployed in an L-shaped mode in upper right corner, consists of the processing unit with glass columns in which the separation takes place; a camera (partially obscurred by Lousma's face) to document the process; and a cryogenic freezer to freeze and store the samples after separation.

  14. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, E.S.; Taylor, J.A.

    1996-03-12

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figs.

  15. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, Edward S. (Ames, IA); Taylor, John A. (Nevada, IA)

    1996-03-12

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

  16. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, Edward S. (Ames, IA); Taylor, John A. (Nevada, IA)

    1994-06-28

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

  17. Positron scattering from vinyl acetate

    NASA Astrophysics Data System (ADS)

    Chiari, L.; Zecca, A.; Blanco, F.; García, G.; Brunger, M. J.

    2014-09-01

    Using a Beer-Lambert attenuation approach, we report measured total cross sections (TCSs) for positron scattering from vinyl acetate (C4H6O2) in the incident positron energy range 0.15-50 eV. In addition, we also report an independent atom model with screening corrected additivity rule computation results for the TCSs, differential and integral elastic cross sections, the positronium formation cross section and inelastic integral cross sections. The energy range of these calculations is 1-1000 eV. While there is a reasonable qualitative correspondence between measurement and calculation for the TCSs, in terms of the energy dependence of those cross sections, the theory was found to be a factor of ˜2 larger in magnitude at the lower energies, even after the measured data were corrected for the forward angle scattering effect.

  18. Neuronal network analysis of serum electrophoresis.

    PubMed Central

    Kratzer, M. A.; Ivandic, B.; Fateh-Moghadam, A.

    1992-01-01

    AIMS: To advise a system of neuronal networks which can classify the densitometric patterns of serum electrophoresis. METHODS: Digitised data containing 83 normal and 132 pathological serum protein electrophoresis patterns were presented to four neuronal networks containing 1900 neurons. Network 1 evaluates the integrated values of the albumin, alpha 1, alpha 2, beta and gamma fractions together with total protein (Biuret method). Networks 2, 3, and 4 analyse the shape of the albumin, beta and gamma fractions. To increase the sensitivity for the detection of monoclonal gammopathies a Fourier transformation was applied to the beta and gamma fractions. RESULTS: After a learning period of 20 minutes (back-propagation learning algorithm) the system was tested with a set of electrophoresis patterns comprising 446 routinely collected samples. It differentiated between physiological and pathological curves with a sensitivity of 97.5% and a specificity of 98.8%, with 86% correct diagnoses. All monoclonal gammopathies were recognised by the Fourier detector. CONCLUSIONS: Neuronal networks could be useful for certain medical uses. Unlike rule based systems, neuronal networks do not have to be programmed but have the capacity to "learn" quickly. PMID:1517463

  19. Extractive fermentation of acetic acid

    SciTech Connect

    Busche, R.M.

    1991-12-31

    In this technoeconomic evaluation of the manufacture of acetic acid by fermentation, the use of the bacterium: Acetobacter suboxydans from the old vinegar process was compared with expected performance of the newer Clostridium thermoaceticum bacterium. Both systems were projected to operate as immobilized cells in a continuous, fluidized bed bioreactor, using solvent extraction to recover the product. Acetobacter metabolizes ethanol aerobically to produce acid at 100 g/L in a low pH medium. This ensures that the product is in the form of a concentrated extractable free acid, rather than as an unextractable salt. Unfortunately, yields from glucose by way of the ethanol fermentation are poor, but near the biological limits of the organisms involved. Conversely, C. thermoaceticum is a thermophilic anaerobe that operates at high fermentation rates on glucose at neutral pH to produce acetate salts directly in substantially quantitative yields. However, it is severely inhibited by product, which restricts concentration to a dilute 20 g/L. An improved Acetobacter system operating with recycled cells at 50 g/L appears capable of producing acid at $0.38/lb, as compared with a $0.29/lb price for synthetic acid. However, this system has only a limited margin for process improvement. The present Clostridium system cannot compete, since the required selling price would be $0.42/lb. However, if the organism could be adapted to tolerate higher product concentrations at acid pH, selling price could be reduced to $0.22/lb, or about 80% of the price of synthetic acid.

  20. Manufacturing Ethyl Acetate From Fermentation Ethanol

    NASA Technical Reports Server (NTRS)

    Rohatgi, Naresh K.; Ingham, John D.

    1991-01-01

    Conceptual process uses dilute product of fermentation instead of concentrated ethanol. Low-concentration ethanol, extracted by vacuum from fermentation tank, and acetic acid constitutes feedstock for catalytic reaction. Product of reaction goes through steps that increases ethyl acetate content to 93 percent by weight. To conserve energy, heat exchangers recycle waste heat to preheat process streams at various points.

  1. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a) Identity. The color additive lead... cosmetics intended for coloring hair on the scalp only, subject to the following restrictions: (1) The amount of the lead acetate in the cosmetic shall be such that the lead content, calculated as Pb,...

  2. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a) Identity. The color additive lead... cosmetics intended for coloring hair on the scalp only, subject to the following restrictions: (1) The amount of the lead acetate in the cosmetic shall be such that the lead content, calculated as Pb,...

  3. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a) Identity. The color additive lead... cosmetics intended for coloring hair on the scalp only, subject to the following restrictions: (1) The amount of the lead acetate in the cosmetic shall be such that the lead content, calculated as Pb,...

  4. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a) Identity. The color additive lead... cosmetics intended for coloring hair on the scalp only, subject to the following restrictions: (1) The amount of the lead acetate in the cosmetic shall be such that the lead content, calculated as Pb,...

  5. 21 CFR 73.2396 - Lead acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2396 Lead acetate. (a) Identity. The color additive lead... cosmetics intended for coloring hair on the scalp only, subject to the following restrictions: (1) The amount of the lead acetate in the cosmetic shall be such that the lead content, calculated as Pb,...

  6. Lipidomic Profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii Reveals Critical Changes in Lipid Composition in Response to Acetic Acid Stress

    PubMed Central

    Riezman, Howard; Olsson, Lisbeth; Bettiga, Maurizio

    2013-01-01

    When using microorganisms as cell factories in the production of bio-based fuels or chemicals from lignocellulosic hydrolysate, inhibitory concentrations of acetic acid, released from the biomass, reduce the production rate. The undissociated form of acetic acid enters the cell by passive diffusion across the lipid bilayer, mediating toxic effects inside the cell. In order to elucidate a possible link between lipid composition and acetic acid stress, the present study presents detailed lipidomic profiling of the major lipid species found in the plasma membrane, including glycerophospholipids, sphingolipids and sterols, in Saccharomyces cerevisiae (CEN.PK 113_7D) and Zygosaccharomyces bailii (CBS7555) cultured with acetic acid. Detailed physiological characterization of the response of the two yeasts to acetic acid has also been performed in aerobic batch cultivations using bioreactors. Physiological characterization revealed, as expected, that Z. bailii is more tolerant to acetic acid than S. cerevisiae. Z. bailii grew at acetic acid concentrations above 24 g L?1, while limited growth of S. cerevisiae was observed after 11 h when cultured with only 12 g L?1 acetic acid. Detailed lipidomic profiling using electrospray ionization, multiple-reaction-monitoring mass spectrometry (ESI-MRM-MS) showed remarkable changes in the glycerophospholipid composition of Z. bailii, including an increase in saturated glycerophospholipids and considerable increases in complex sphingolipids in both S. cerevisiae (IPC 6.2×, MIPC 9.1×, M(IP)2C 2.2×) and Z. bailii (IPC 4.9×, MIPC 2.7×, M(IP)2C 2.7×), when cultured with acetic acid. In addition, the basal level of complex sphingolipids was significantly higher in Z. bailii than in S. cerevisiae, further emphasizing the proposed link between lipid saturation, high sphingolipid levels and acetic acid tolerance. The results also suggest that acetic acid tolerance is associated with the ability of a given strain to generate large rearrangements in its lipid profile. PMID:24023914

  7. Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

    DOEpatents

    Zhang, Jian-Shi; Giometti, C.S.; Tollaksen, S.L.

    1987-09-04

    After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a dc power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, so that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. 8 figs.

  8. Development of a pH sensor based on a nanostructured filter adding pH-sensitive fluorescent dye for detecting acetic acid in photovoltaic modules

    NASA Astrophysics Data System (ADS)

    Asaka, Takashi; Itayama, Tomohiro; Nagasaki, Hideaki; Iwami, Kentaro; Yamamoto, Chizuko; Hara, Yukiko; Masuda, Atsushi; Umeda, Norihiro

    2015-08-01

    Acetic acid formed via the hydrolysis of ethylene vinyl acetate (EVA) as an encapsulant in photovoltaic (PV) modules causes a decrease in the conversion efficiency of such modules by grid corrosion. Here, a nondestructive and simple optical method for evaluating the condition of PV modules is proposed. This method uses a dual-wavelength pH-sensitive fluorescent dye to detect acetic acid in PV modules using a change in pH. The change in pH induced by the formation of acetic acid is detected by the change in the ratio of the fluorescent intensities of two peaks of the dye. A pH-sensitive fluorescent dye showed sensitivity for small amounts of acetic acid such as that produced from EVA. Furthermore, a membrane filter dyed with a pH-sensitive fluorescent dye was confirmed to detect acetic acid in aged EVA after a damp-heat test (85 °C, 85%) for 5000 h in PV modules.

  9. NUCLEAR MEMBRANES FROM MAMMALIAN LIVER

    PubMed Central

    Franke, Werner W.; Deumling, Barbara; Ermen, Baerbel; Jarasch, Ernst-Dieter; Kleinig, Hans

    1970-01-01

    Nuclear membranes were isolated from rat and pig liver by sonication of highly purified nuclear fractions and subsequent removal of adhering nucleoproteins in a high salt medium. The fractions were examined in the electron microscope by both negative staining and thin sectioning techniques and were found to consist of nuclear envelope fragments of widely varying sizes. Nuclear pore complex constituents still could frequently be recognized. The chemical composition of the nuclear membrane fractions was determined and compared with those of microsomal fractions prepared in parallel. For total nuclei as well as for nuclear membranes and microsomes, various enzyme activities were studied. The results indicate that a similarity exists between both fractions of cytomembranes, nuclear envelope, and endoplasmic reticulum, with respect to their RNA:protein ratio and their content of polar and nonpolar lipids. Both membranous fractions had many proteins in common including some membrane-bound enzymes. Activities in Mg-ATPase and the two examined cytochrome reductases were of the same order of magnitude. The content of cytochrome b5 as well as of P-450 was markedly lower in the nuclear membranes. The nuclear membranes were found to have a higher buoyant density and to be richer in protein. The glucose-6-phosphatase and Na-K-ATPase activities in the nuclear membrane fraction were very low. In the gel electrophoresis, in addition to many common protein bands, some characteristic ones for either microsomal or nuclear membranous material were detected. Significant small amounts of DNA and RNA were found to remain closely associated with the nuclear envelope fragments. Our findings indicate that nuclear and endoplasmic reticulum membranes which are known to be in morphological continuity have, besides a far-reaching similarity, some characteristic differences. PMID:4317731

  10. Biological Hydrogen Production Using a Membrane Bioreactor

    E-print Network

    ; methanogen inhibition; cross-flow membrane; Clostridia- ceae; Flexibacteraceae INTRODUCTION Hydrogen). Hydrogen gas can be produced directly through fermentative routes at high concentrations (approximately 60 by a known biochemical route used by bacteria is 4 moles of hydrogen when acetic acid is produced (Gottschalk

  11. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 2010-04-01 false Vitamin A acetate. 582.5933 Section 582...AS SAFE Nutrients and/or Dietary Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of...

  12. The fluid mechanics of continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.

    1990-01-01

    The overall objective is to establish theoretically and confirm experimentally the ultimate capabilities of continuous flow electrophoresis chambers operating in an environment essentially free of particle sedimentation and buoyancy. The efforts are devoted to: (1) studying the effects of particle concentration on sample conductivity and dielectric constant. The dielectric constant and conductivity were identified as playing crucial roles in the behavior of the sample and on the resolving power and throughput of continuous flow devices; and (2) improving the extant mathematical models to predict flow fields and particle trajectories in continuous flow electrophoresis. A dielectric spectrometer was designed and built to measure the complex dielectric constant of a colloidal dispersion as a function of frequency between 500 Hz and 200 kHz. The real part of the signal can be related to the sample's conductivity and the imaginary part to its dielectric constant. Measurements of the dielectric constants of several different dispersions disclosed that the dielectric constants of dilute systems of the sort encountered in particle electrophoresis are much larger than would be expected based on the extant theory. Experiments were carried out to show that, in many cases, this behavior is due to the presence of a filamentary structure of small hairs on the particle surface. A technique for producing electrokinetically ideal synthetic latex particles by heat treating was developed. Given the ubiquitous nature of hairy surfaces with both cells and synthetic particles, it was deemed necessary to develop a theory to explain their behavior. A theory for electrophoretic mobility of hairy particles was developed. Finally, the extant computer programs for predicting the structure of electro-osmotically driven flows were extended to encompass flow channels with variable wall mobilities.

  13. Pulsed field gel electrophoresis for dairy propionibacteria.

    PubMed

    Chuat, Victoria; de Freitas, Rosangela; Dalmasso, Marion

    2015-01-01

    Pulsed field gel electrophoresis (PFGE) is a technique using alternating electric fields to migrate high molecular weight DNA fragments with a high resolution. This method consists of the digestion of bacterial chromosomal DNA with rare-cutting restriction enzymes and in applying an alternating electrical current between spatially distinct pairs of electrodes. DNA molecules migrate at different speeds according to the size of the fragments. Among other things, this technique is considered as the "gold standard" for genotyping, genetic fingerprinting, epidemiological studies, genome size estimation, and studying radiation-induced DNA damage and repair. This chapter describes a PFGE method that can be used to differentiate dairy propionibacteria. PMID:25862063

  14. Fluid mechanics of continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.; Ostrach, S.

    1978-01-01

    The following aspects of continuous flow electrophoresis were studied: (1) flow and temperature fields; (2) hydrodynamic stability; (3) separation efficiency, and (4) characteristics of wide gap chambers (the SPAR apparatus). Simplified mathematical models were developed so as to furnish a basis for understanding the phenomena and comparison of different chambers and operating conditions. Studies of the hydrodynamic stability disclosed that a wide gap chamber may be particularly sensitive to axial temperature variations which could be due to uneven heating or cooling. The mathematical model of the separation process includes effects due to the axial velocity, electro-osmotic cross flow and electrophoretic migration, all including the effects of temperature dependent properties.

  15. Microfabricated capillary array electrophoresis device and method

    DOEpatents

    Simpson, Peter C.; Mathies, Richard A.; Woolley, Adam T.

    2004-06-15

    A capillary array electrophoresis (CAE) micro-plate with an array of separation channels connected to an array of sample reservoirs on the plate. The sample reservoirs are organized into one or more sample injectors. One or more waste reservoirs are provided to collect wastes from reservoirs in each of the sample injectors. Additionally, a cathode reservoir is also multiplexed with one or more separation channels. To complete the electrical path, an anode reservoir which is common to some or all separation channels is also provided on the micro-plate. Moreover, the channel layout keeps the distance from the anode to each of the cathodes approximately constant.

  16. Microfabricated capillary array electrophoresis device and method

    DOEpatents

    Simpson, Peter C. (Oakland, CA); Mathies, Richard A. (Moraga, CA); Woolley, Adam T. (Belmont, MA)

    2000-01-01

    A capillary array electrophoresis (CAE) micro-plate with an array of separation channels connected to an array of sample reservoirs on the plate. The sample reservoirs are organized into one or more sample injectors. One or more waste reservoirs are provided to collect wastes from reservoirs in each of the sample injectors. Additionally, a cathode reservoir is also multiplexed with one or more separation channels. To complete the electrical path, an anode reservoir which is common to some or all separation channels is also provided on the micro-plate. Moreover, the channel layout keeps the distance from the anode to each of the cathodes approximately constant.

  17. A new approach to scaling up electrophoresis

    SciTech Connect

    Tarnopolsky, Y.; Roman, M.; Brown, P.R.

    1993-01-01

    Free Flow Electrophoresis (FFE) has been utilized for the separation of proteins and cells for many years, and has evolved into the most promising method of continuous separation. One of the major drawbacks inherent with FFE, however, is the thermal convection due to Joule heating which occurs whenever current is passed through a conducting solution. To provide efficient heat dissipation, the size of FFE units is restricted, which limits sample throughput. A new type of FFE design, which internally cools the separation unit by passing water through capillary tubes, has been developed and tested. Results of separations of dyes are presented, using a bed {1/4} inch thick which maintains efficient cooling.

  18. Hybrid slab-microchannel gel electrophoresis system

    DOEpatents

    Balch, Joseph W. (Livermore, CA); Carrano, Anthony V. (Livermore, CA); Davidson, James C. (Livermore, CA); Koo, Jackson C. (San Ramon, CA)

    1998-01-01

    A hybrid slab-microchannel gel electrophoresis system. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate.

  19. Electrohydrodynamic effects in continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H.; Snyder, R. S.; Roberts, G. O.; Baygents, J. C.

    1991-01-01

    We demonstrate experimentally and theoretically the importance of electrohydrodynamic (EHD) flows in continuous-flow electrophoresis (CFE) separations. These flows are associated with variations in the conductivity or dielectric constant, and are quadratic in the field strength. They appear to be the main cause of extraneous and undesired flows in CFE which have degraded separation performance and have until now not been explained. We discuss the importance of EHD flows relative to other effects. We also describe possible techniques for reducing the associated degradation of CFE separations.

  20. A centrifugal method for the evaluation of polymer membranes for reverse osmosis

    NASA Technical Reports Server (NTRS)

    Hollahan, J. R.; Wydeven, T.; Mccullough, R. P.

    1973-01-01

    A rapid and simple method employing the laboratory centrifuge shows promise for evaluation of membrane performance during reverse osmosis. Results are presented for cellulose acetate membranes for rejection of salt and urea dissolved solids. Implications of the study are to rapid screening of membrane performance, use in laboratories with limited facilities, and possible space waste water purification.

  1. Recent advances in microchip electrophoresis for amino acid analysis.

    PubMed

    Ou, Gaozhi; Feng, Xiaojun; Du, Wei; Liu, Xin; Liu, Bi-Feng

    2013-10-01

    With the maturation of microfluidic technologies, microchip electrophoresis has been widely employed for amino acid analysis owing to its advantages of low sample consumption, reduced analysis time, high throughput, and potential for integration and automation. In this article, we review the recent progress in amino acid analysis using microchip electrophoresis during the period from 2007 to 2012. Innovations in microchip materials, surface modification, sample introduction, microchip electrophoresis, and detection methods are documented, as well as nascent applications of amino acid analysis in single-cell analysis, microdialysis sampling, food analysis, and extraterrestrial exploration. Without doubt, more applications of microchip electrophoresis in amino acid analysis may be expected soon. PMID:23436170

  2. Distinct Effects of Sorbic Acid and Acetic Acid on the Electrophysiology and Metabolism of Bacillus subtilis

    PubMed Central

    van Beilen, J. W. A.; Teixeira de Mattos, M. J.; Hellingwerf, K. J.

    2014-01-01

    Sorbic acid and acetic acid are among the weak organic acid preservatives most commonly used to improve the microbiological stability of foods. They have similar pKa values, but sorbic acid is a far more potent preservative. Weak organic acids are most effective at low pH. Under these circumstances, they are assumed to diffuse across the membrane as neutral undissociated acids. We show here that the level of initial intracellular acidification depends on the concentration of undissociated acid and less on the nature of the acid. Recovery of the internal pH depends on the presence of an energy source, but acidification of the cytosol causes a decrease in glucose flux. Furthermore, sorbic acid is a more potent uncoupler of the membrane potential than acetic acid. Together these effects may also slow the rate of ATP synthesis significantly and may thus (partially) explain sorbic acid's effectiveness. PMID:25038097

  3. The pharmacology of nomegestrol acetate.

    PubMed

    Ruan, Xiangyan; Seeger, Harald; Mueck, Alfred O

    2012-04-01

    Nomegestrol acetate (NOMAC) is a 19-norprogesterone derivative with high biological activity at the progesterone receptor, a weak anti-androgenic effect, but with no binding to estrogen, glucocorticoid or mineralocorticoid receptors. At dosages of 1.5mg/day or more, NOMAC effectively suppresses gonadotropic activity and ovulation in women of reproductive age. Hemostasis, lipids and carbohydrate metabolism remain largely unchanged. In normal and cancerous human breast cells, NOMAC has shown favorable effects on estrogen metabolism. Like natural progesterone (but in contrast to some other synthetic progestogens), it does not appear stimulate the proliferation of cancerous breast cells. While there has been some experience of the use of NOMAC in combination with estrogens as a hormone replacement therapy, most of the data on the compound are reported in the context of its inclusion as a component of a new contraceptive pill comprising 2.5mg NOMAC combined with 1.5mg estradiol. Because of its strong endometrial efficacy, and due to its high antigonadotropic activity and long elimination half-life (about 50h), the contraceptive efficacy of the new pill is maintained even when dosages are missed. Furthermore, for the first time with a monophasic 24/4 regimen containing estradiol, cyclical stability can be achieved comparable with that obtained using pills containing ethinyl estradiol and progestogens like levonorgestrel or drospirenone. The addition of NOMAC to estradiol means that the beneficial effects of estrogen are not lost, which is of especial importance in relation to the cardiovascular system. On the basis both of its pharmacology and of studies performed during the development of the NOMAC/estradiol pill, involving some 4000 women in total, good long-term tolerability can be expected for NOMAC, although its safety profile is still to be fully ascertained, as the clinical endpoint studies are yet to be completed. PMID:22364709

  4. Micro-injector for capillary electrophoresis.

    PubMed

    Sáiz, Jorge; Koenka, Israel Joel; García-Ruiz, Carmen; Müller, Beat; Chwalek, Thomas; Hauser, Peter C

    2015-08-01

    A novel micro-injector for capillary electrophoresis for the handling of samples with volumes down to as little as 300 nL was designed and built in our laboratory for analyses in which the available volume is a limitation. The sample is placed into a small cavity located directly in front of the separation capillary, and the injection is then carried out automatically by controlled pressurization of the chamber with compressed air. The system also allows automated flushing of the injection chamber as well as of the capillary. In a trial with a capillary electrophoresis system with contactless conductivity detector, employing a capillary of 25 ?m diameter, the results showed good stability of migration times and peak areas. To illustrate the technique, the fast separation of five inorganic cations (Na(+) , K(+) , NH4 (+) , Ca(2+) , and Mg(2+) ) was set up. This could be achieved in less than 3 min, with good limits of detection (10 ?M) and linear ranges (between about 10 and 1000 ?M). The system was demonstrated for the determination of the inorganic cations in porewater samples of a lake sediment core. PMID:25752271

  5. Floating resistivity detector for microchip electrophoresis.

    PubMed

    Tay, Elaine Teng Teng; Law, Wai Siang; Sim, Steven Poh Chuen; Feng, Huatao; Zhao, Jian Hong; Li, Sam Fong Yau

    2007-12-01

    A newly developed conductivity detector, the floating resistivity detector (FRD), for microchip electrophoresis was introduced in this work. The detector design permits decoupling of the detection circuit from the high separation voltage without compromising separation efficiency. This greatly simplifies the integration of microchip electrophoresis systems. Its method of detection relies on platinum electrodes being dipped in two buffer-filled branched detection probe reservoirs on the microchip device. In this way, analytes passing through the detection window will not pass through and subsequently adsorb onto the electrodes, alleviating problems of electrode fouling due to analyte contamination and surface reactions. A customized microchip design was proposed and optimized stepwise for the new FRD system. Each branched detection probe was determined to be 4.50 mm long with a 0.075 mm detection window gap between them. The distance between the detection window and buffer waste reservoir was determined to be 1.50 mm. The optimized microchip design was subsequently used in the analysis of four groups of analytes - inorganic cations, amino acids, aminoglycosides antibiotics, and biomarkers. Based on the preliminary results obtained, the detection limits were in the range of 0.4-0.7 mg/L for the inorganic cations and 1.5-15 mg/L for the amino compounds. PMID:18072226

  6. Fractionation of mineral species by electrophoresis

    NASA Technical Reports Server (NTRS)

    Dunning, J. D.; Herren, B. J.; Tipps, R. W.; Snyder, R. S.

    1982-01-01

    The fractionation of fine-grained aggregates into their major components is a problem in many scientific areas including earth and planetary science. Electrophoresis, the transport of electrically charged particles, immersed in a suspension medium, by a direct current field (Bier, 1959), was employed in this study as a means of separating simulated lunar soil into its constituent minerals. In these tests, conducted in a static analytical cylindrical microelectrophoresis apparatus, samples of simulated lunar soil and samples of pure mineral constituents were placed in the chamber; the electrophoretic mobilities of the lunar soil and the individual mineral constituents were measured. In most of the suspension buffers employed separability was indicated, on the basis of differences in mobility, for all the constituent mineral species except ilmenite and pyroxene, which were not efficiently separable in any of the buffers. Although only a few suspension media were employed, the success of this initial study suggests that electrophoresis may be an important mineral fractionation option in fine-grained aggregate processing.

  7. Gel Electrophoresis of Gold-DNA Nanoconjugates

    DOE PAGESBeta

    Pellegrino, T.; Sperling, R. A.; Alivisatos, A. P.; Parak, W. J.

    2007-01-01

    Gold-DNA conjugates were investigated in detail by a comprehensive gel electrophoresis study based on 1200 gels. A controlled number of single-stranded DNA of different length was attached specifically via thiol-Au bonds to phosphine-stabilized colloidal gold nanoparticles. Alternatively, the surface of the gold particles was saturated with single stranded DNA of different length either specifically via thiol-Au bonds or by nonspecific adsorption. From the experimentally determined electrophoretic mobilities, estimates for the effective diameters of the gold-DNA conjugates were derived by applying two different data treatment approaches. The first method is based on making a calibration curve for the relation between effectivemore »diameters and mobilities with gold nanoparticles of known diameter. The second method is based on Ferguson analysis which uses gold nanoparticles of known diameter as reference database. Our study shows that effective diameters derived from gel electrophoresis measurements are affected with a high error bar as the determined values strongly depend on the method of evaluation, though relative changes in size upon binding of molecules can be detected with high precision. Furthermore, in this study, the specific attachment of DNA via gold-thiol bonds to Au nanoparticles is compared to nonspecific adsorption of DNA. Also, the maximum number of DNA molecules that can be bound per particle was determined.« less

  8. Removal of bisphenol A (BPA) from water by various nanofiltration (NF) and reverse osmosis (RO) membranes.

    PubMed

    Yüksel, Suna; Kabay, Nalan; Yüksel, Mithat

    2013-12-15

    The removal of an endocrine disrupting compound, bisphenol A (BPA), from model solutions by selected nanofiltration (NF) and reverse osmosis (RO) membranes was studied. The commercially available membranes NF 90, NF 270, XLE BWRO, BW 30 (Dow FilmTech), CE BWRO and AD SWRO (GE Osmonics) were used to compare their performances for BPA removal. The water permeability coefficients, rejection of BPA and permeate flux values were calculated for all membranes used. No significant changes in their BPA removal were observed for all tight polyamide based NF and RO membranes tested except for loose NF 270 membrane. The polyamide based membranes exhibited much better performance than cellulose acetate membrane for BPA removal. Almost a complete rejection (? 98%) for BPA was obtained with three polyamide based RO membranes (BW 30, XLE BWRO and AD SWRO). But cellulose acetate based CE BWRO membrane offered a low and variable (10-40%) rejection for BPA. PMID:23731784

  9. 21 CFR 522.1073 - Gonadorelin acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS § 522.1073 Gonadorelin acetate. (a) Specifications. Each...

  10. Concentrating aqueous acetate solutions with tertiary amines 

    E-print Network

    Lee, Champion

    1993-01-01

    was originally applied to water desalination in which water was extracted from aqueous sodium chloride solutions. Here, we explore its potential to recover acetate produced via fermentation. At 40C 55C, which corresponds to typical fen-fermentation temperatures...

  11. Acetic Acid Off Gassing in Clamshell Enclosures

    E-print Network

    Brewer, Allison

    2013-01-01

    . This presentation will investigate the use of acid detection strips (A-D strips) to study acetic acid off gassing occurring in custom-made, cloth covered book boxes constructed and used by conservators in research libraries....

  12. Fragrance material review on 4-methylbenzyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 4-methylbenzyl acetate when used as a fragrance ingredient is presented. 4-Methylbenzyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 4-methylbenzyl acetate were evaluated, then summarized, and includes: physical properties, skin irritation, skin sensitization, and elicitation data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414643

  13. Fragrance material review on 3-phenylpropyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 3-phenylpropyl acetate when used as a fragrance ingredient is presented. 3-Phenylpropyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 3-phenylpropyl acetate were evaluated, then summarized, and includes: physical properties, acute toxicity, skin irritation, skin sensitization, and toxicokinetics data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al., 2012 for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414651

  14. Fragrance material review on anisyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of anisyl acetate when used as a fragrance ingredient is presented. Anisyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for anisyl acetate were evaluated, then summarized, and includes: physical properties, skin irritation, skin sensitization, elicitation, and phototoxicity data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al., 2012 for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414654

  15. [carbonyl-11 C]Benzyl acetate: Automated

    E-print Network

    Pike, Victor W.

    Fujita,a Osamu Inoue,b Robert B. Innis,a and Victor W. Pikea [carbonyl-11 C]Benzyl acetate ([11 C]1) has, so increasing the difficulty of their use in small-scale automated radiochemistry. Organolithium

  16. Indium acetate toxicity in male reproductive system in rats.

    PubMed

    Lee, Kuo-Hsin; Chen, Hsiu-Ling; Leung, Chung-Man; Chen, Hsin-Pao; Hsu, Ping-Chi

    2016-01-01

    Indium, a rare earth metal characterized by high plasticity, corrosion resistance, and a low melting point, is widely used in the electronics industry, but has been reported to be an environmental pollutant and a health hazard. We designed a study to investigate the effects of subacute exposure of indium compounds on male reproductive function. Twelve-week old male Sprague-Dawley rats were randomly divided into test and control groups, and received weekly intraperitoneal injections of indium acetate (1.5 mg/kg body weight) and normal saline, respectively, for 8 weeks. Serum indium levels, cauda epididymal sperm count, motility, morphology, chromatin DNA structure, mitochondrial membrane potential, oxidative stress, and testis DNA content were investigated. The indium acetate-treated group showed significant reproductive toxicity, as well as an increased percentage of sperm morphology abnormality, chromatin integrity damage, and superoxide anion generation. Furthermore, positive correlations among sperm morphology abnormalities, chromatin DNA damage, and superoxide anion generation were also noted. The results of this study demonstrated the toxic effect of subacute low-dose indium exposure during the period of sexual maturation on male reproductive function in adulthood, through an increase in oxidative stress and sperm chromatin DNA damage during spermiogenesis, in a rodent model. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 68-76, 2016. PMID:25044390

  17. Androgen dynamics in vitro in the human prostate gland. Effect of cyproterone and cyproterone acetate

    PubMed Central

    Giorgi, Eleonora P.; Shirley, I. M.; Grant, J. K.; Stewart, Joan C.

    1973-01-01

    Hyperplastic and adenocarcinomatous human prostatic tissue was superfused in vitro with radioactively labelled androst-4-ene-3,17-dione, testosterone and 5?-dihydrotestosterone (17?-hydroxy-5?-androstan-3-one), with and without addition of the anti-androgens cyproterone and cyproterone acetate. Cyproterone competitively inhibited the entry of the androgens into the majority of the tissues, whereas cyproterone acetate increased this entry. These findings indicated that transport of androstenedione, testosterone and 5?-dihydrotestosterone into prostatic tissue is performed by a specific mechanism, possibly involving a carrier situated in the cell membrane. The extent of metabolism of the three androgens was also modified: formation of 5?-dihydrotestosterone from testosterone, and of the latter from androstenedione, was decreased by cyproterone and increased by the acetate. Acetate was more effective than cyproterone in decreasing the `uptake' of the perfused androgens by the tissue; at the same time, it increased the androgen clearance from the tissue. As cyproterone acetate is the more potent of the two anti-androgens, the possibility that these findings in vitro are related to the different anti-androgenic potency exhibited by the two compounds in vivo is discussed. `Uptake' of the two anti-androgens and the response to their action on androgen dynamics were similar in adenocarcinomatous and hyperplastic glands. PMID:4125095

  18. Activation of methyl acetate on Pd(111)

    NASA Astrophysics Data System (ADS)

    Xu, Lijun; Xu, Ye

    2010-06-01

    The adsorption and activation of methyl acetate (CH 3COOCH 3), one of the simplest carboxylic esters, on Pd(111) have been studied using self-consistent periodic density functional theory calculations. Methyl acetate adsorbs weakly through the carbonyl oxygen. Its activation occurs via dehydrogenation, instead of direct C-O bond dissociation, on clean Pd(111): It is much more difficult to dissociate the C-O bonds ( Ea ? 2.0 eV for the carbonyl and acetate-methyl bonds; Ea = 1.0 eV for the acetyl-methoxy bond) than to dissociate the C-H bonds to produce enolate (CH 2COOCH 3; Ea = 0.74 eV) or methylene acetate (CH 3COOCH 2; Ea = 0.82 eV). The barriers for C-H and C-O bond dissociation are directly calculated for enolate and methylene acetate, and estimated for further dehydrogenated derivatives (CH 3COOCH, CH 2COOCH 2, and CHCOOCH 3) based on the Brønsted-Evans-Polanyi linear energy relations formed by the calculated steps. The enolate pathway leads to successive dehydrogenation to CCOOCH 3, whereas methylene acetate readily dissociates to yield acetyl. The selectivity for dissociating the acyl-alkoxy C-O bond, which is desired for alcohol formation, is therefore fundamentally limited by the facility of dehydrogenation under vacuum/low-pressure conditions on Pd(111).

  19. Analysis of Central European Corydalis species by nonaqueous capillary electrophoresis-electrospray ion trap mass spectrometry.

    PubMed

    Sturm, Sonja; Seger, Christoph; Stuppner, Hermann

    2007-08-01

    In the presented study, Central European Corydalis species, namely C. cava, C. intermedia, C. pumila, and C. solida were investigated by nonaqueous capillary electrophoresis-electrospray ion trap mass spectrometry (NACE-ESI-MS) utilizing an electrolyte consisting of 50 mM ammonium acetate, 1 M acetic acid and 10% methanol in acetonitrile, applying a separation voltage of 30 and 20 kV for 1 s for injection. Isopropanol-water (1:1) was used as sheath liquid at a flow-rate of 3 microl/min. Peak assignment was assisted by multistage-ESI-mass spectrometry (ESI-MSn). The optimized method was fully validated (RSD inter- and intraday < 5%, LOD < 2.3 microg/ml, LOQ < 27.6 microg/ml, recovery rates > 98.8%) and subsequently applied for the qualitative and quantitative determination of isoquinoline alkaloids in single plant tubers (sample size < 0.5 g) of the four Central European Corydalis species, each of them showing a characteristic and unique alkaloid pattern. Application of a principal component analysis (PCA) to the complete dataset of 39 analytes and 79 samples allowed the identification of 8 analytes responsible for lot discrimination. Hierarchical cluster analysis and descriptive statistical methods were used to confirm the findings of the explorative PCA. PMID:17395190

  20. Nanofiltration of rhodium tris(triphenylphosphine) catalyst in ethyl acetate solution

    NASA Astrophysics Data System (ADS)

    Shaharun, Maizatul S.; Mustafa, Ahmad K.; Taha, Mohd F.

    2012-09-01

    Solvent resistant nanofiltration (SRNF) using polymer membranes has recently received enhanced attention due to the search for cleaner and more energy-efficient technologies. The large size of the rhodium tris(triphenylphosphine) [HRh(CO)(PPh3)3] catalyst (>400 Da) - relative to other components of the hydroformylation reaction provides the opportunity for a membrane separation based on retention of the catalyst species while permeating the solvent. The compatibility of the solvent-polyimide membrane (DuraMem{trade mark, serif} 200 and DuraMem{trade mark, serif} 500) combinations was assessed in terms of the membrane stability in solvent plus non-zero solvent flux at 2.0 MPa. Good HRh(CO)(PPh3)3 rejection (>0.95) and solvent fluxes of 9.9 L/m2?h1 at 2.0 MPa were obtained in the catalyst-ethyl acetate-DuraMem 500 system. The effect of pressure and catalyst concentration on the solvent flux and catalyst rejection was conducted on the catalyst-ethyl acetate-membrane systems. Increasing pressure substantially improved both solvent flux and catalyst rejection, while increasing catalyst concentration was found to be beneficial in terms of substantial increases in catalyst rejection without significantly affecting solvent flux.

  1. Modification of gel architecture and TBE/TAE buffer composition to minimize heating during agarose gel electrophoresis

    PubMed Central

    Sanderson, Brian A.; Araki, Naoko; Lilley, Jennifer L.; Guerrero, Gilberto; Lewis, L. Kevin

    2014-01-01

    Agarose gel electrophoresis of DNA and RNA is routinely performed using buffers containing either Tris, acetate and EDTA (TAE) or Tris, borate and EDTA (TBE). Gels are run at a low, constant voltage (~ 10 V/cm) to minimize current and asymmetric heating effects, which can induce band artifacts and poor resolution. In this study, alterations of gel structure and conductive media composition were analyzed to identify factors causing higher electrical currents during horizontal slab gel electrophoresis. Current was reduced when thinner gels and smaller chamber buffer volumes were used, but was not influenced by agarose concentration or the presence of ethidium bromide. Current was strongly dependent upon the amount and type of EDTA used and on the concentrations of the major acid-base components of each buffer. Interestingly, resolution and the mobilities of circular versus linear plasmid DNAs were also affected by the chemical form and amount of EDTA. With appropriate modifications to gel structure and buffer constituents, electrophoresis could be performed at high voltages (20–25 V/cm), reducing run times by up to 3-fold. The most striking improvements were observed with small DNAs and RNAs (10 – 100 bp): high voltages and short run times produced sharper bands and higher resolution. PMID:24637158

  2. Membrane Extraction for Detoxification of Biomass Hydrolysates

    SciTech Connect

    Grzenia, D. L.; Schell, D. J.; Wickramasinghe, S. R.

    2012-05-01

    Membrane extraction was used for the removal of sulfuric acid, acetic acid, 5-hydroxymethyl furfural and furfural from corn stover hydrolyzed with dilute sulfuric acid. Microporous polypropylene hollow fiber membranes were used. The organic extractant consisted of 15% Alamine 336 in: octanol, a 50:50 mixture of oleyl alcohol:octanol or oleyl alcohol. Rapid removal of sulfuric acid, 5-hydroxymethyl and furfural was observed. The rate of acetic acid removal decreased as the pH of the hydrolysate increased. Regeneration of the organic extractant was achieved by back extraction into an aqueous phase containing NaOH and ethanol. A cleaning protocol consisting of flushing the hydrolysate compartment with NaOH and the organic phase compartment with pure organic phase enabled regeneration and reuse of the module. Ethanol yields from hydrolysates detoxified by membrane extraction using 15% Alamine 336 in oleyl alcohol were about 10% higher than those from hydrolysates detoxified using ammonium hydroxide treatment.

  3. Membrane extraction for detoxification of biomass hydrolysates.

    PubMed

    Grzenia, David L; Schell, Daniel J; Wickramasinghe, S Ranil

    2012-05-01

    Membrane extraction was used for the removal of sulfuric acid, acetic acid, 5-hydroxymethyl furfural and furfural from corn stover hydrolyzed with dilute sulfuric acid. Microporous polypropylene hollow fiber membranes were used. The organic extractant consisted of 15% Alamine 336 in: octanol, a 50:50 mixture of oleyl alcohol:octanol or oleyl alcohol. Rapid removal of sulfuric acid, 5-hydroxymethyl and furfural was observed. The rate of acetic acid removal decreased as the pH of the hydrolysate increased. Regeneration of the organic extractant was achieved by back extraction into an aqueous phase containing NaOH and ethanol. A cleaning protocol consisting of flushing the hydrolysate compartment with NaOH and the organic phase compartment with pure organic phase enabled regeneration and reuse of the module. Ethanol yields from hydrolysates detoxified by membrane extraction using 15% Alamine 336 in oleyl alcohol were about 10% higher than those from hydrolysates detoxified using ammonium hydroxide treatment. PMID:22361069

  4. Western blotting of basic proteins after nondenaturing electrophoresis in acid conditions using the PhastSystem.

    PubMed

    Davril, M; Ducourouble, M P; Van-Seuningen, I

    1993-09-01

    Electroblotting of basic proteins was performed from minigels after electrophoresis, under nondenaturing acidic conditions, by using the automated PhastSystem. Depending on the molecular masses of the proteins to be studied, various precast gel media were chosen. The transfer membranes with various types (nitrocellulose and polyvinylidene difluoride) and pore sizes (0.45 and 0.2 micron) were chosen accordingly. For the semidry electric transfer, a simple, discontinuous two-buffer system was used. The anode solution contained 0.3 M Tris, pH 10.4, and the cathode solution, 40 mM 6-amino-n-hexanoic acid, pH 7.6, with 20% v/v methanol each. The addition of 0.1% sodium dodecyl sulfate (SDS) in the cathode solution facilitated the elution of proteins from the gels and directed the migration of the negative SDS-protein complexes towards the anode membranes. The transfer conditions following native polyacrylamide gel electrophoresis allowed the visualization of basic proteins, with molecular weights ranging from 29,000 to 5,000, for which isoforms could be resolved and which retained their biological properties. PMID:8223396

  5. Gel Electrophoresis on a Budget to Dye for

    ERIC Educational Resources Information Center

    Yu, Julie H.

    2010-01-01

    Gel electrophoresis is one of the most important tools used in molecular biology and has facilitated the entire field of genetic engineering by enabling the separation of nucleic acids and proteins. However, commercial electrophoresis kits can cost up to $800 for each setup, which is cost prohibitive for most classroom budgets. This article…

  6. Inexpensive and Safe DNA Gel Electrophoresis Using Household Materials

    ERIC Educational Resources Information Center

    Ens, S.; Olson, A. B.; Dudley, C.; Ross, N. D., III; Siddiqi, A. A.; Umoh, K. M.; Schneegurt, M. A.

    2012-01-01

    Gel electrophoresis is the single most important molecular biology technique and it is central to life sciences research, but it is often too expensive for the secondary science classroom or homeschoolers. A simple safe low-cost procedure is described here that uses household materials to construct and run DNA gel electrophoresis. Plastic…

  7. Two-Dimensional Gel Electrophoresis in Platelet Proteomics Research

    E-print Network

    23 Two-Dimensional Gel Electrophoresis in Platelet Proteomics Research Ángel García Summary Proteomics technology allows a comprehensive and efficient analysis of the proteome and has become electrophoresis (2-DE) in proteomics and its application to platelet research. 2-DE separates proteins according

  8. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Electrophoresis apparatus for clinical use. 862... SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An...

  9. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Electrophoresis apparatus for clinical use. 862.2485 Section 862.2485 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An...

  10. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Electrophoresis apparatus for clinical use. 862.2485 Section 862.2485 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An...

  11. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Electrophoresis apparatus for clinical use. 862.2485 Section 862.2485 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An...

  12. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Electrophoresis apparatus for clinical use. 862.2485 Section 862.2485 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An...

  13. Polymeric matrices for DNA sequencing by capillary electrophoresis

    E-print Network

    Barron, Annelise E.

    materials that have been tested for DNA sequencing by capillary elec- trophoresis (CE). HypothesizedPolymeric matrices for DNA sequencing by capillary electrophoresis We review the wide range of polymeric materials that have been employed for DNA sequencing separations by capillary electrophoresis

  14. Analysis of the Microbial Community in an Acidic Hollow-Fiber Membrane Biofilm Reactor (Hf-MBfR) Used for the Biological Conversion of Carbon Dioxide to Methane

    PubMed Central

    Jeon, Byoung Seung; Choi, Okkyoung; Kim, Hyun Wook; Um, Youngsoon; Lee, Dong-Hoon; Sang, Byoung-In

    2015-01-01

    Hydrogenotrophic methanogens can use gaseous substrates, such as H2 and CO2, in CH4 production. H2 gas is used to reduce CO2. We have successfully operated a hollow-fiber membrane biofilm reactor (Hf-MBfR) for stable and continuous CH4 production from CO2 and H2. CO2 and H2 were diffused into the culture medium through the membrane without bubble formation in the Hf-MBfR, which was operated at pH 4.5–5.5 over 70 days. Focusing on the presence of hydrogenotrophic methanogens, we analyzed the structure of the microbial community in the reactor. Denaturing gradient gel electrophoresis (DGGE) was conducted with bacterial and archaeal 16S rDNA primers. Real-time qPCR was used to track changes in the community composition of methanogens over the course of operation. Finally, the microbial community and its diversity at the time of maximum CH4 production were analyzed by pyrosequencing methods. Genus Methanobacterium, related to hydrogenotrophic methanogens, dominated the microbial community, but acetate consumption by bacteria, such as unclassified Clostridium sp., restricted the development of acetoclastic methanogens in the acidic CH4 production process. The results show that acidic operation of a CH4 production reactor without any pH adjustment inhibited acetogenic growth and enriched the hydrogenotrophic methanogens, decreasing the growth of acetoclastic methanogens. PMID:26694756

  15. Investigations of the inhibitory effects of tocopherol (vitamin E) on free radical deterioration of cellular membranes

    NASA Technical Reports Server (NTRS)

    Richardson, D.

    1975-01-01

    The inhibitory effects are investigated of d,1-alpha-tocopherol and d,1-alpha-tocopheryl acetate on the free radical deterioration of cellular membranes. The level of toxicity of d,1-alpha-tocopherol and d,1-alpha-tocopheryl acetate in mice is determined.

  16. Direct Detection of the Acetate-forming Activity of the Enzyme Acetate Kinase

    PubMed Central

    Fowler, Matthew L.; Ingram-Smith, Cheryl J.; Smith, Kerry S.

    2011-01-01

    Acetate kinase, a member of the acetate and sugar kinase-Hsp70-actin (ASKHA) enzyme superfamily1-5, is responsible for the reversible phosphorylation of acetate to acetyl phosphate utilizing ATP as a substrate. Acetate kinases are ubiquitous in the Bacteria, found in one genus of Archaea, and are also present in microbes of the Eukarya6. The most well characterized acetate kinase is that from the methane-producing archaeon Methanosarcina thermophila7-14. An acetate kinase which can only utilize PPi but not ATP in the acetyl phosphate-forming direction has been isolated from Entamoeba histolytica, the causative agent of amoebic dysentery, and has thus far only been found in this genus15,16. In the direction of acetyl phosphate formation, acetate kinase activity is typically measured using the hydroxamate assay, first described by Lipmann17-20, a coupled assay in which conversion of ATP to ADP is coupled to oxidation of NADH to NAD+ by the enzymes pyruvate kinase and lactate dehydrogenase21,22, or an assay measuring release of inorganic phosphate after reaction of the acetyl phosphate product with hydroxylamine23. Activity in the opposite, acetate-forming direction is measured by coupling ATP formation from ADP to the reduction of NADP+ to NADPH by the enzymes hexokinase and glucose 6-phosphate dehydrogenase24. Here we describe a method for the detection of acetate kinase activity in the direction of acetate formation that does not require coupling enzymes, but is instead based on direct determination of acetyl phosphate consumption. After the enzymatic reaction, remaining acetyl phosphate is converted to a ferric hydroxamate complex that can be measured spectrophotometrically, as for the hydroxamate assay. Thus, unlike the standard coupled assay for this direction that is dependent on the production of ATP from ADP, this direct assay can be used for acetate kinases that produce ATP or PPi. PMID:22214984

  17. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, Richard D.; Udseth, Harold R.; Olivares, Jose A.

    1994-10-18

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5-100 KVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., .+-.2-8 KVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit.

  18. Novel absorption detection techniques for capillary electrophoresis

    SciTech Connect

    Xue, Y.

    1994-07-27

    Capillary electrophoresis (CE) has emerged as one of the most versatile separation methods. However, efficient separation is not sufficient unless coupled to adequate detection. The narrow inner diameter (I.D.) of the capillary column raises a big challenge to detection methods. For UV-vis absorption detection, the concentration sensitivity is only at the {mu}M level. Most commercial CE instruments are equipped with incoherent UV-vis lamps. Low-brightness, instability and inefficient coupling of the light source with the capillary limit the further improvement of UV-vis absorption detection in CE. The goals of this research have been to show the utility of laser-based absorption detection. The approaches involve: on-column double-beam laser absorption detection and its application to the detection of small ions and proteins, and absorption detection with the bubble-shaped flow cell.

  19. Hybrid slab-microchannel gel electrophoresis system

    DOEpatents

    Balch, J.W.; Carrano, A.V.; Davidson, J.C.; Koo, J.C.

    1998-05-05

    A hybrid slab-microchannel gel electrophoresis system is described. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate. 4 figs.

  20. Flow structure in continuous flow electrophoresis chambers

    NASA Technical Reports Server (NTRS)

    Deiber, J. A.; Saville, D. A.

    1982-01-01

    There are at least two ways that hydrodynamic processes can limit continiuous flow electrophoresis. One arises from the sensitivity of the flow to small temerature gradients, especially at low flow rates and power levels. This sensitivity can be suppressed, at least in principle, by providing a carefully tailored, stabilizing temperature gradient in the cooling system that surrounds the flow channel. At higher power levels another limitation arises due to a restructuring of the main flow. This restructuring is caused by buoyancy, which is in turn affected by the electro-osmotic crossflow. Approximate solutions to appropriate partial differential equations have been computed by finite difference methods. One set of results is described here to illustrate the strong coupling between the structure of the main (axial) flow and the electro-osmotic flow.

  1. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, Richard P. (Richland, WA); Udseth, Harold R. (Richland, WA); Olivares, Jose A. (North Augusta, SC)

    1989-01-01

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5-100 KVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., .+-.2-8 KVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit.

  2. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, R.P.; Udseth, H.R.; Olivares, J.A.

    1989-12-05

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5--100 kVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., [+-]2--8 kVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit. 21 figs.

  3. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, R.D.; Udseth, H.R.; Olivares, J.A.

    1994-10-18

    A system and method for analyzing molecular constituents of a composition sample include: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5--100 kVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g.,{+-}2--8 kVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit. 21 figs.

  4. Acetylation of Starch with Vinyl Acetate in Imidazolium Ionic Liquids and Characterization of Acetate Distribution

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Starch was acetylated with vinyl acetate in different 1-butyl-3-methylimidazolium (BMIM) salts as solvent in effort to produce starches with different acetylation patterns. Overall degree of substitution was much higher for basic anions such as acetate and dicyanimide (dca) than for neutral anions ...

  5. Tested Demonstrations: Buffer Capacity of Various Acetic Acid-Sodium Acetate Systems: A Lecture Experiment.

    ERIC Educational Resources Information Center

    Donahue, Craig J.; Panek, Mary G.

    1985-01-01

    Background information and procedures are provided for a lecture experiment which uses indicators to illustrate the concept of differing buffer capacities by titrating acetic acid/sodium acetate buffers with 1.0 molar hydrochloric acid and 1.0 molar sodium hydroxide. A table with data used to plot the titration curve is included. (JN)

  6. Evaluation of different protein extraction methods for banana (Musa spp.) root proteome analysis by two-dimensional electrophoresis.

    PubMed

    Vaganan, M Mayil; Sarumathi, S; Nandakumar, A; Ravi, I; Mustaffa, M M

    2015-02-01

    Four protocols viz., the trichloroacetic acid-acetone (TCA), phenol-ammonium acetate (PAA), phenol/SDS-ammonium acetate (PSA) and trisbase-acetone (TBA) were evaluated with modifications for protein extraction from banana (Grand Naine) roots, considered as recalcitrant tissues for proteomic analysis. The two-dimensional electrophoresis (2-DE) separated proteins were compared based on protein yield, number of resolved proteins, sum of spot quantity, average spot intensity and proteins resolved in 4-7 pI range. The PAA protocol yielded more proteins (0.89 mg/g of tissues) and protein spots (584) in 2-DE gel than TCA and other protocols. Also, the PAA protocol was superior in terms of sum of total spot quantity and average spot intensity than TCA and other protocols, suggesting phenol as extractant and ammonium acetate as precipitant of proteins were the most suitable for banana rooteomics analysis by 2-DE. In addition, 1:3 ratios of root tissue to extraction buffer and overnight protein precipitation were most efficient to obtain maximum protein yield. PMID:26040117

  7. Phosphorylation of membrane proteins in erythrocytes treated with lead.

    PubMed Central

    Belloni-Olivi, L; Annadata, M; Goldstein, G W; Bressler, J P

    1996-01-01

    In immature rat microvessels, endothelial cells and glioma cells, exposure to lead results in an increase in the level of protein kinase C in membranes. In this paper we have extended these studies to human erythrocytes and, in addition, studied the phosphorylation of membrane proteins. A significant increase in the phosphorylation of membrane cytoskeletal proteins of molecular mass 120, 80, 52 and 45 kDa was observed in human erythrocytes treated for 60 min with lead acetate at concentrations greater than 100 nM. These same proteins were phosphorylated when erythrocytes were treated for 10 min with 50 nM phorbol 12-myristate 13-acetate (PMA). Similarly, protein kinase C activity was elevated and an increase in the amount of protein kinase C-alpha was observed in membranes from erythrocytes exposed to concentrations of lead acetate above 100 nM. No changes, however, in the activities of cAMP-dependent protein kinase, protein phosphatases I and IIA or casein kinase were observed. Phosphorylation of these membrane proteins stimulated by lead acetate or by PMA was not observed in erythrocytes depleted of protein kinase C by a 72-h treatment with 500 nM phorbol 12,13-dibutyrate. Finally, no changes in the levels of calcium or diacylglycerol were observed in erythrocytes stimulated with 100 nM lead acetate. These results indicate that, in erythrocytes, lead acetate stimulates the phosphorylation of membrane cytoskeletal proteins by a mechanism dependent on protein kinase C. Since levels of calcium or diacylglycerols did not increase, it appears that lead may activate the enzyme by a direct interaction. PMID:8615806

  8. Cloning, sequence analysis, and hyperexpression of the genes encoding phosphotransacetylase and acetate kinase from Methanosarcina thermophila.

    PubMed Central

    Latimer, M T; Ferry, J G

    1993-01-01

    The genes for the acetate-activating enzymes, acetate kinase and phosphotransacetylase (ack and pta), from Methanosarcina thermophila TM-1 were cloned and sequenced. Both genes are present in only one copy per genome, with the pta gene adjacent to and upstream of the ack gene. Consensus archaeal promoter sequences are found upstream of the pta coding region. The pta and ack genes encode predicted polypeptides with molecular masses of 35,198 and 44,482 Da, respectively. A hydropathy plot of the deduced phosphotransacetylase sequence indicates that it is a hydrophobic polypeptides; however, no membrane-spanning domains are evident. Comparison of the amino acid sequences deduced from the M. thermophila and Escherichia coli ack genes indicate similar subunit molecular weights and 44% identity (60% similarity). The comparison also revealed the presence of several conserved arginine, cysteine, and glutamic acid residues. Arginine, cysteine, and glutamic acid residues have previously been implicated at or near the active site of the E. coli acetate kinase. The pta and ack genes were hyperexpressed in E. coli, and the overproduced enzymes were purified to homogeneity with specific activities higher than those of the enzymes previously purified from M. thermophila. The overproduced phosphotransacetylase and acetate kinase migrated at molecular masses of 37,000 and 42,000 Da, respectively. The activity of the acetate kinase is optimal at 65 degrees C and is protected from thermal inactivation by ATP. Diethylpyrocarbonate and phenylglyoxal inhibited acetate kinase activity in a manner consistent with the presence of histidine and arginine residues at or near the active site; however, the thiol-directed reagents 5,5'-dithiobis (2-nitrobenzoic acid) and N-ethylmaleimide were ineffective. Images PMID:8226623

  9. A novel approach to pseudopodia proteomics: excimer laser etching, two-dimensional difference gel electrophoresis, and confocal imaging

    PubMed Central

    Mimae, Takahiro; Ito, Akihiko; Hagiyama, Man; Nakanishi, Jun; Hosokawa, Yoichiroh; Okada, Morihito; Murakami, Yoshinori; Kondo, Tadashi

    2014-01-01

    Pseudopodia are actin-rich ventral cellular protrusions shown to facilitate the migration and metastasis of tumor cells. Here, we present a novel approach to perform pseudopodia proteomics. Tumor cells growing on porous membranes extend pseudopodia into the membrane pores. In our method, cell bodies are removed by horizontal ablation at the basal cell surface with the excimer laser while pseudopodia are left in the membrane pores. For protein expression profiling, whole cell and pseudopodia proteins are extracted with a lysis buffer, labeled with highly sensitive fluorescent dyes, and separated by two-dimensional gel electrophoresis. Proteins with unique expression patterns in pseudopodia are identified by mass spectrometry. The effects of the identified proteins on pseudopodia formation are evaluated by measuring the pseudopodia length in cancer cells with genetically modified expression of target proteins using confocal imaging. This protocol allows global identification of pseudopodia proteins and evaluation of their functional significance in pseudopodia formation within one month. PMID:25309719

  10. Extended length microchannels for high density high throughput electrophoresis systems

    DOEpatents

    Davidson, James C. (Livermore, CA); Balch, Joseph W. (Livermore, CA)

    2000-01-01

    High throughput electrophoresis systems which provide extended well-to-read distances on smaller substrates, thus compacting the overall systems. The electrophoresis systems utilize a high density array of microchannels for electrophoresis analysis with extended read lengths. The microchannel geometry can be used individually or in conjunction to increase the effective length of a separation channel while minimally impacting the packing density of channels. One embodiment uses sinusoidal microchannels, while another embodiment uses plural microchannels interconnected by a via. The extended channel systems can be applied to virtually any type of channel confined chromatography.

  11. A systematic study of field inversion gel electrophoresis.

    PubMed Central

    Heller, C; Pohl, F M

    1989-01-01

    The mobilities of oligomers of phage lambda DNA and of yeast chromosomes in agarose gels during field inversion gel electrophoresis (FIGE) were measured at different pulse times and electric fields. Also the ratios between forward and backward pulse times and/or field gradients were varied. The problem of 'band inversion' during FIGE, leading to an ambiguity in the mobility of large DNA fragments, was solved by using two dimensional gel electrophoresis with different parameters in the first and second dimension. The results are compared with those obtained with other pulsed electrophoresis systems and with a theoretical model. Images PMID:2528121

  12. DNA sequencing using fluorescence background electroblotting membrane

    DOEpatents

    Caldwell, K.D.; Chu, T.J.; Pitt, W.G.

    1992-05-12

    A method for the multiplex sequencing on DNA is disclosed which comprises the electroblotting or specific base terminated DNA fragments, which have been resolved by gel electrophoresis, onto the surface of a neutral non-aromatic polymeric microporous membrane exhibiting low background fluorescence which has been surface modified to contain amino groups. Polypropylene membranes are preferably and the introduction of amino groups is accomplished by subjecting the membrane to radio or microwave frequency plasma discharge in the presence of an aminating agent, preferably ammonia. The membrane, containing physically adsorbed DNA fragments on its surface after the electroblotting, is then treated with crosslinking means such as UV radiation or a glutaraldehyde spray to chemically bind the DNA fragments to the membrane through amino groups contained on the surface. The DNA fragments chemically bound to the membrane are subjected to hybridization probing with a tagged probe specific to the sequence of the DNA fragments. The tagging may be by either fluorophores or radioisotopes. The tagged probes hybridized to the target DNA fragments are detected and read by laser induced fluorescence detection or autoradiograms. The use of aminated low fluorescent background membranes allows the use of fluorescent detection and reading even when the available amount of DNA to be sequenced is small. The DNA bound to the membranes may be reprobed numerous times. No Drawings

  13. Membrane Processes.

    PubMed

    Pellegrin, Marie-Laure; Sadler, Mary E; Greiner, Anthony D; Aguinaldo, Jorge; Min, Kyungnan; Zhang, Kai; Arabi, Sara; Burbano, Marie S; Kent, Fraser; Shoaf, Robert

    2015-10-01

    This review, for literature published in 2014, contains information related to membrane processes for municipal and industrial applications. This review is a subsection of the Treatment Systems section of the annual Water Environment Federation literature review and covers the following topics: pretreatment, membrane bioreactor (MBR) configuration, design, nutrient removal, operation, industrial treatment, fixed film and anaerobic membrane systems, reuse, microconstituents removal, membrane technology advances, membrane fouling, and modeling. Other sub-sections of the Treatment Systems section that might relate to this literature review include: Biological Fixed-Film Systems, Activated Sludge and Other Aerobic Suspended Culture Processes, Anaerobic Processes, Water Reclamation and Reuse. The following sections might also have related information on membrane processes: Industrial Wastes, Hazardous Wastes, and Fate and Effects of Pollutants. PMID:26420079

  14. Multicomponent membranes

    DOEpatents

    Kulprathipanja, Santi (Hoffman Estates, IL); Kulkarni, Sudhir S. (Hoffman Estates, IL); Funk, Edward W. (Highland Park, IL)

    1988-01-01

    A multicomponent membrane which may be used for separating various components which are present in a fluid feed mixture comprises a mixture of a plasticizer such as a glycol and an organic polymer cast upon a porous organic polymer support. The membrane may be prepared by casting an emulsion or a solution of the plasticizer and polymer on the porous support, evaporating the solvent and recovering the membrane after curing.

  15. Microfab-less Microfluidic Capillary Electrophoresis Devices

    PubMed Central

    Segato, Thiago P.; Bhakta, Samir A.; Gordon, Matthew; Carrilho, Emanuel; Willis, Peter A.; Jiao, Hong; Garcia, Carlos D.

    2013-01-01

    Compared to conventional bench-top instruments, microfluidic devices possess advantageous characteristics including great portability potential, reduced analysis time (minutes), and relatively inexpensive production, putting them on the forefront of modern analytical chemistry. Fabrication of these devices, however, often involves polymeric materials with less-than-ideal surface properties, specific instrumentation, and cumbersome fabrication procedures. In order to overcome such drawbacks, a new hybrid platform is proposed. The platform is centered on the use of 5 interconnecting microfluidic components that serve as the injector or reservoirs. These plastic units are interconnected using standard capillary tubing, enabling in-channel detection by a wide variety of standard techniques, including capacitively-coupled contactless conductivity detection (C4D). Due to the minimum impact on the separation efficiency, the plastic microfluidic components used for the experiments discussed herein were fabricated using an inexpensive engraving tool and standard Plexiglas. The presented approach (named 52-platform) offers a previously unseen versatility: enabling the assembly of the platform within minutes using capillary tubing that differs in length, diameter, or material. The advantages of the proposed design are demonstrated by performing the analysis of inorganic cations by capillary electrophoresis on soil samples from the Atacama Desert. PMID:23585815

  16. Integrated polymerase chain reaction/electrophoresis instrument

    DOEpatents

    Andresen, Brian D. (Livermore, CA)

    2000-01-01

    A new approach and instrument for field identification of micro-organisms and DNA fragments using a small and disposable device containing integrated polymerase chain reaction (PCR) enzymatic reaction wells, attached capillary electrophoresis (CE) channels, detectors, and read-out all on/in a small hand-held package. The analysis instrument may be made inexpensively, for example, of plastic, and thus is disposable, which minimizes cross contamination and the potential for false positive identification between samples. In addition, it is designed for multiple users with individual applications. The integrated PCR/CE is manufactured by the PCR well and CE channels are "stamped" into plastic depressions where conductive coatings are made in the wells and ends of the CE microchannels to carry voltage and current to heat the PCR reaction mixtures and simultaneously draw DNA bands up the CE channels. Light is transmitted through the instrument at appropriate points and detects PCR bands and identifies DNA fragments by size (retention time) and quantifies each by the amount of light generated as each phototransistor positioned below each CE channel detects a passing band. The instrument is so compact that at least 100 PCR/CE reactions/analyses can be performed easily on one detection device.

  17. Strongly nonlinear waves in capillary electrophoresis

    PubMed Central

    Chen, Zhen; Ghosal, Sandip

    2012-01-01

    In capillary electrophoresis, sample ions migrate along a micro-capillary filled with a background electrolyte under the influence of an applied electric field. If the sample concentration is sufficiently high, the electrical conductivity in the sample zone could differ significantly from the background. Under such conditions, the local migration velocity of sample ions becomes concentration dependent resulting in a nonlinear wave that exhibits shock like features. If the nonlinearity is weak, the sample concentration profile, under certain simplifying assumptions, can be shown to obey Burgers’ equation (S. Ghosal and Z. Chen Bull. Math. Biol. 2010 72(8), pg. 2047) which has an exact analytical solution for arbitrary initial condition. In this paper, we use a numerical method to study the problem in the more general case where the sample concentration is not small in comparison to the concentration of background ions. In the case of low concentrations, the numerical results agree with the weakly nonlinear theory presented earlier, but at high concentrations, the wave evolves in a way that is qualitatively different. PMID:23004798

  18. Mini-electrochemical detector for microchip electrophoresis.

    PubMed

    Jiang, Lei; Lu, Yao; Dai, Zhongpeng; Xie, Minhao; Lin, Bingcheng

    2005-09-01

    This paper presents the development of a mini-electrochemical detector for microchip electrophoresis. The small size (3.6 x 5.0 cm2, W x L) of the detector is compatible with the dimension of the microchip. The use of universal serial bus (USB) ports facilitates installation and use of the detector, miniaturizes the detector, and makes it ideal for lab-on-a-chip applications. A fixed 10 M ohm feedback resistance was chosen to convert current of the working electrode to voltage with second gain of 1, 2, 4, 8, 16, 32, 64 and 128 for small signal detection instead of adopting selectable feedback resistance. Special attention has been paid to the power support circuitry and printed circuit board (PCB) design in order to obtain good performance in such a miniature size. The working electrode potential could be varied over a range of +/-2.5 V with a resolution of 0.01 mV. The detection current ranges from -0.3 x 10(-7) A to 2.5 x 10(-7) A and the noise is lower than 1 pA. The analytical performance of the new system was demonstrated by the detection of epinephrine using an integrated PDMS/glass microchip with detection limit of 2.1 microM (S/N = 3). PMID:16100576

  19. Polyethylene-supported polyvinylidene fluoride-cellulose acetate butyrate blended polymer electrolyte for lithium ion battery

    NASA Astrophysics Data System (ADS)

    Liu, Jiansheng; Li, Weishan; Zuo, Xiaoxi; Liu, Shengqi; Li, Zhao

    2013-03-01

    The polyethylene (PE)-supported polymer membranes based on the blended polyvinylidene fluoride (PVDF) and cellulose acetate butyrate (CAB) are prepared for gel polymer electrolyte (GPE) of lithium ion battery. The performances of the prepared membranes and the resulting GPEs are investigated by scanning electron microscopy, electrochemical impedance spectroscopy, linear potential sweep, and charge-discharge test. The effect of the ratio of PVDF to CAB on the performance of the prepared membranes is considered. It is found that the GPE based on the blended polymer with PVDF:CAB = 2:1 (in weight) has the largest ionic conductivity (2.48 × 10-3 S cm-1) and shows good compatibility with anode and cathode of lithium ion battery. The LiCoO2/graphite battery using this GPE exhibits superior cyclic stability at room temperature, storage performance at elevated temperature, and rate performance.

  20. Fragrance material review on 1,1-dimethyl-2-phenylethyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 1,1-dimethyl-2-phenylethyl acetate when used as a fragrance ingredient is presented. 1,1-Dimethyl-2-phenylethyl acetate is a member of the fragrance structural group Aryl Alkyl Alcohol Simple Acid Esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an Aryl Alkyl Alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to 1,1-dimethyl-2-phenylethyl acetate and is not intended as a stand-alone document. Available data were evaluated, then summarized, and includes: physical properties; acute toxicity; skin irritation; mucous membrane (eye) irritation; skin sensitization; elicitation; and toxicokinetics data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22445843

  1. Cathepsin D protects colorectal cancer cells from acetate-induced apoptosis through autophagy-independent degradation of damaged mitochondria

    PubMed Central

    Oliveira, C S F; Pereira, H; Alves, S; Castro, L; Baltazar, F; Chaves, S R; Preto, A; Côrte-Real, M

    2015-01-01

    Acetate is a short-chain fatty acid secreted by Propionibacteria from the human intestine, known to induce mitochondrial apoptotic death in colorectal cancer (CRC) cells. We previously established that acetate also induces lysosome membrane permeabilization in CRC cells, associated with release of the lysosomal protease cathepsin D (CatD), which has a well-established role in the mitochondrial apoptotic cascade. Unexpectedly, we showed that CatD has an antiapoptotic role in this process, as pepstatin A (a CatD inhibitor) increased acetate-induced apoptosis. These results mimicked our previous data in the yeast system showing that acetic acid activates a mitochondria-dependent apoptosis process associated with vacuolar membrane permeabilization and release of the vacuolar protease Pep4p, ortholog of mammalian CatD. Indeed, this protease was required for cell survival in a manner dependent on its catalytic activity and for efficient mitochondrial degradation independently of autophagy. In this study, we therefore assessed the role of CatD in acetate-induced mitochondrial alterations. We found that, similar to acetic acid in yeast, acetate-induced apoptosis is not associated with autophagy induction in CRC cells. Moreover, inhibition of CatD with small interfering RNA or pepstatin A enhanced apoptosis associated with higher mitochondrial dysfunction and increased mitochondrial mass. This effect seems to be specific, as inhibition of CatB and CatL with E-64d had no effect, nor were these proteases significantly released to the cytosol during acetate-induced apoptosis. Using yeast cells, we further show that the role of Pep4p in mitochondrial degradation depends on its protease activity and is complemented by CatD, indicating that this mechanism is conserved. In summary, the clues provided by the yeast model unveiled a novel CatD function in the degradation of damaged mitochondria when autophagy is impaired, which protects CRC cells from acetate-induced apoptosis. CatD inhibitors could therefore enhance acetate-mediated cancer cell death, presenting a novel strategy for prevention or therapy of CRC. PMID:26086961

  2. Free zone electrophoresis simulation of static column electrophoresis in microgravity on shuttle flight STS-3

    NASA Technical Reports Server (NTRS)

    Todd, P. W.; Hjerten, S.

    1985-01-01

    Experiments were designed to replicate, as closely as possible in 1-G, the conditions of the STS-3 red blood cell (RBC) experiments. Free zone electrophoresis was the method of choice, since it minimizes the role of gravity in cell migration. The physical conditions of the STS-3 experiments were used, and human and rabbit RBC's fixed by the same method were the test particles. The effects of cell concentration, electroosmotic mobility, and sample composition were tested in order to seek explanations for the STS-3 results and to provide data on cell concentration effects for future zero-G separation on the continuous-flow zero-G electrophoretics separator.

  3. Process for the preparation of vinyl acetate

    DOEpatents

    Tustin, G.C.; Zoeller, J.R.; Depew, L.S.

    1998-02-17

    This invention pertains to the preparation of vinyl acetate by contacting within a contact zone a mixture of ketene and acetaldehyde with an acid catalyst at about one bar pressure and between about 85 and 200 C and removing the reaction products from the contact zone.

  4. Process for the preparation of vinyl acetate

    DOEpatents

    Tustin, Gerald Charles (Kingsport, TN); Zoeller, Joseph Robert (Kingsport, TN); Depew, Leslie Sharon (Kingsport, TN)

    1998-01-01

    This invention pertains to the preparation of vinyl acetate by contacting within a contact zone a mixture of ketene and acetaldehyde with an acid catalyst at about one bar pressure and between about 85.degree. and 200.degree. C. and removing the reaction products from the contact zone.

  5. Synthesis of Cellulose Acetate from Cotton Byproducts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton burr and cottonseed hull are relatively inexpensive cotton byproducts. In an effort to derive greater value out of these natural renewable materials, we have succeeded in converting part of them into cellulose acetate without prior chemical breakdown or physical separation of cellulose, ligni...

  6. Fermentative biohydrogen production from lactate and acetate.

    PubMed

    Wu, Chao-Wei; Whang, Liang-Ming; Cheng, Hai-Hsuan; Chan, Kan-Chi

    2012-06-01

    In this study, a continuous-flow stirred tank reactor (CSTR) fed with lactate and acetate was operated to enrich hydrogen-producing bacteria. By varying the influent substrate concentrations and hydraulic retention times (HRT), the volumetric loading rate (VLR) of 55.64 kg-COD/m(3)/day seemed to be optimum for this enriched culture for fermentative hydrogen production from lactate and acetate. The results of batch experiments confirmed that the enriched culture tended to fulfill the e(-) equiv requirement for cell growth at a lower VLR condition (21.77 kg-COD/m(3)/day), while it could largely distribute the e(-) equiv for hydrogen production at a higher VLR condition. However, a maximum lactate/acetate concentration allowed for enriching this culture existed, especially at a lower HRT condition in which wash-out can be an issue for this enriched culture. Finally, the results of cloning and sequencing indicated that Clostridium tyrobutyricum was considered the major hydrogen-producing bacteria in the CSTR fed with lactate and acetate. PMID:22318084

  7. Advanced Colloids Experiment (ACE-T1)

    NASA Technical Reports Server (NTRS)

    Meyer, William V.; Sicker, Ron; Brown, Dan; Eustace, John

    2015-01-01

    Increment 45 - 46 Science Symposium presentation of Advanced Colloids Experiment (ACE-T1) to RPO. The purpose of this event is for Principal Investigators to present their science objectives, testing approach, and measurement methods to agency scientists, managers, and other investigators.

  8. Single molecule analysis of DNA electrophoresis in microdevices

    E-print Network

    Randall, Greg C

    2006-01-01

    Given that current electrophoresis technology is inadequate for mapping large O[100 kilobasepair] DNA, several promising lab-on-chip designs for DNA mapping have been recently proposed that require either 1) a DNA molecule ...

  9. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...use. (a) Identification. An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including plasma proteins, lipoproteins, enzymes, and hemoglobulins on the basis of their net charge...

  10. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...use. (a) Identification. An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including plasma proteins, lipoproteins, enzymes, and hemoglobulins on the basis of their net charge...

  11. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...use. (a) Identification. An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including plasma proteins, lipoproteins, enzymes, and hemoglobulins on the basis of their net charge...

  12. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including plasma proteins, lipoproteins, enzymes, and hemoglobulins on the basis of their net charge in specified buffered media. This...

  13. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including plasma proteins, lipoproteins, enzymes, and hemoglobulins on the basis of their net charge in specified buffered media. This...

  14. Non-linear electrophoresis of ideally polarizable particles

    E-print Network

    Chan, Wai Hong Ronald

    2014-01-01

    This thesis investigates the non-linear regime of electrophoresis, in particular the variation of electrophoretic velocity with electric field at high field strengths. Known theoretical approaches to the problem accounting ...

  15. CAPILLARY ELECTROPHORESIS FOR THE CHARACTERIZATION OF HUMIC SUBSTANCES

    EPA Science Inventory

    The potential of high performance capillary electrophoresis (HPCE), especially in the free solution mode (FSCE), is demonstrated for the analysis/characterization of environmental humic substances (HUS). he very high efficiency of HPCE separations allows the production of electro...

  16. Microchannel DNA Sequencing by End-Labelled Free Solution Electrophoresis

    SciTech Connect

    Barron, A.

    2005-09-29

    The further development of End-Labeled Free-Solution Electrophoresis will greatly simplify DNA separation and sequencing on microfluidic devices. The development and optimization of drag-tags is critical to the success of this research.

  17. A model for sample stacking in microcapillary DNA electrophoresis

    E-print Network

    Srivastava, Alok Kumar, 1967-

    2002-01-01

    Sanger's method of chain termination is the method of choice in DNA sequencing, where electrophoresis is used to separate the different sized DNA. In the past decade, microfabricated capillary devices have been developed ...

  18. A New Electrophoresis Technique to Seperate Microsatellite Alleles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional agarose and polyacrylamide gel electrophoresis have been used commonly for microsatellite (simple sequence repeats, SSRs) analysis, but they are labor- intensive and not always able to provide accurate sizes for different alleles. Capillary sequencers provide automated analysis and accur...

  19. Effects of non-thermal plasma on the electrical properties of an erythrocyte membrane

    NASA Astrophysics Data System (ADS)

    Lee, Jin Young; Baik, Ku Youn; Kim, Tae Soo; Lim, Jaekwan; Uhm, Han S.; Choi, Eun Ha

    2015-09-01

    Non-thermal plasma is used here for membrane oxidation and permeabilization in which the electrical properties of an erythrocyte membrane are investigated after treatments. The zeta potential as measured by electrophoresis shows the increased negativity of the membrane surface potential (?s). The secondary electron emission coefficient ( ?) measured by a focused ion beam shows a decrease in the dipole potential (?d) of lipid molecules. The voltage-sensitive fluorescent intensity as measured by flow cytometry shows a decrease in the trans-membrane potential (??) through the lipid bilayer membrane. These results allow us to take a step forward to unveil the complex events occurring in plasma-treated cells.

  20. Calmodulin-binding proteins in chromaffin cell plasma membranes.

    PubMed

    Fournier, S; Trifaró, J M

    1988-11-01

    Calmodulin-binding proteins present in chromaffin cell plasma membranes were isolated and directly compared with calmodulin-binding proteins present in chromaffin granule membranes. Chromaffin cell plasma membranes were prepared using Cytodex 1 microcarriers. Marker enzyme studies on this preparation showed a nine- to 10-fold plasma membrane enrichment over cell homogenates and a low contamination of these plasma membranes by subcellular organelles. Plasma membranes prepared in this manner were solubilized with Triton X-100 and applied to a calmodulin-affinity column in the presence of calcium. Several major calmodulin-binding proteins (240, 105, and 65 kilodaltons) were eluted by an EGTA-containing buffer. 125I-Calmodulin overlay experiments on nitrocellulose sheets containing both chromaffin plasma and granule membranes showed that these two membranes have several calmodulin-binding proteins in common (65, 60, 53, and 50 kilodaltons), as well as unique calmodulin-binding proteins (34 kilodaltons in granule membranes and 240 and 160 kilodaltons in plasma membranes). The 65-kilodalton calmodulin-binding protein present in both membrane types was shown to consist of two isoforms (pI 6.0 and 6.2) by two-dimensional gel electrophoresis. Previous experiments from our laboratory, using two monoclonal antibodies (mAb 30 and mAb 48) specific for a rat brain synaptic vesicle membrane protein (p65), showed that the monoclonal antibodies reacted with a 65-kilodalton calmodulin-binding protein present in at least three neurosecretory vesicles (chromaffin granules, neurohypophyseal granules, and rat brain synaptic vesicles). When these monoclonal antibodies were tested on chromaffin cell plasma membranes and calmodulin-binding proteins isolated from these membranes, they recognized a 65-kilodalton protein. These results indicate that an immunologically identical calmodulin-binding protein is expressed in both chromaffin granule membranes (as well as other secretory vesicle membranes) and chromaffin cell plasma membranes, thus suggesting a possible role for this protein in granule/plasma membrane interaction. PMID:3171592

  1. Capillary Electrophoresis as a Fundamental Probe of Polymer Dynamics

    E-print Network

    George D. J. Phillies

    2011-02-25

    Capillary electrophoresis has long been been recognized as a powerful analytic tool. Here it is demonstrated that the same capillary electrophoretic experiments also reveal dynamic properties of the polymer solutions being used as the support medium. The dependence of the electrophoretic mobility on the size of the probe and the properties of the matrix polymers shows a unity of behavior between electrophoresis and other methods of studying polymer properties.

  2. Novel separation and detection methods of DNA fragments in electrophoresis

    SciTech Connect

    Chan, K.C.

    1992-01-01

    A charge-coupled device (CCD) based electrophoresis system was developed. The system allowed non-destructive, sensitive, and on-line detection of native DNA in slab-gel electrophoresis via ultraviolet absorption measurement. The detection limit of double-stranded DNA fragment was 5 ng per band. Since the amount of DNA used in this experiment was typical, the CCD-based system could be readily implemented in molecular biology. Gel-filled and non-gel sieving capillary electrophoresis (CE) was developed for rapid and efficient separation of double-stranded DNA fragments. For the gel-filled CE separation a new gel matrix, the HydroLink gel (HL), was used. The HL capillary gel was easier to cast than the polyacrylamide capillary gel. For the non-gel separation, a GC capillary was used as the separation chamber, and cellulose additive was included in the electrophoresis as the sieving medium. Indirect fluorometry was applied in non-gel and gel electrophoresis for the detection of DNA fragments. This method allowed nondestructive and on-line detection of DNA during electrophoresis. The amount of DNA used with this method was comparable to those obtained with absorption measurement.

  3. Novel separation and detection methods of DNA fragments in electrophoresis

    SciTech Connect

    Chan, King Cheung.

    1993-01-27

    A charge-coupled device (CCD) based electrophoresis system was developed. The system allowed non-destructive, sensitive, and on-line detection of native DNA in slab-gel electrophoresis via ultraviolet absorption measurement. The detection limit of double-stranded DNA fragment was 5 ng per band. Since the amount of DNA used in this experiment was typical, the CCD-based system could be readily implemented in molecular biology. Gel-filled and non-gel sieving capillary electrophoresis was developed for rapid and efficient separation of double-stranded DNA fragments. For the gel-filled CE separation a new gel matrix, the HydroLink gel (HL), was used. The HL capillary gel was easier to cast than the polyacrylamide capillary gel. For the non-gel separation, a GC capillary was used as the separation chamber, and cellulose additive was included in the electrophoresis as the sieving medium. Indirect fluorometry was applied in non-gel and gel electrophoresis for the detection of DNA fragments. This method allowed non-destructive and on-line detection of DNA during electrophoresis. The amount of DNA used with this method was comparable to those obtained with absorption measurement.

  4. Muscle protein analysis by two-dimensional gel electrophoresis

    SciTech Connect

    Giometti, C.S.

    1982-01-01

    Two-dimensional electrophoresis of muscle proteins has provided valuable new information concerning the heterogeneity of some of the major contractile proteins, alterations in the protein population of developing muscle fibers during various stages of myogenesis, and protein aberrations that correlate with muscle diseases. As with all electrophoretic techniques, careful attention must be paid to the preparation of samples and the selection of reagents to be used for the protein separations. Two-dimensional electrophoresis is the obvious method of choice when analysis of protein mixtures is required. The routine clinical application of two-dimensional electrophoresis to analysis of muscle tissue remains to be demonstrated. However, methods of sample preparation for two-dimensional electrophoresis compatible with existing clinical procedures have been described, and the equipment for multiple analyses is available. As protein abnormalities related to human myopathy are detected through the use of two-dimensional electrophoresis as a research tool, useful clinical markers of specific myopathic processes will be found. The preliminary work on muscle protein analysis by two-dimensional electrophoresis described in this review has begun a new approach to the enigma of human muscle disease.

  5. Phenyl Acetate Preparation from Phenol and Acetic Acid: Reassessment of a Common Textbook Misconception.

    ERIC Educational Resources Information Center

    Hocking, M. B.

    1980-01-01

    Reassesses a common textbook misconception that "...phenols cannot be esterified directly." Results of experiments are discussed and data tables provided of an effective method for the direct preparation of phenyl acetate. (CS)

  6. Characterization of the Acetate Binding Pocket in the Methanosarcina thermophila Acetate Kinase

    PubMed Central

    Ingram-Smith, Cheryl; Gorrell, Andrea; Lawrence, Sarah H.; Iyer, Prabha; Smith, Kerry; Ferry, James G.

    2005-01-01

    Acetate kinase catalyzes the reversible magnesium-dependent synthesis of acetyl phosphate by transfer of the ATP ?-phosphoryl group to acetate. Inspection of the crystal structure of the Methanosarcina thermophila enzyme containing only ADP revealed a solvent-accessible hydrophobic pocket formed by residues Val93, Leu122, Phe179, and Pro232 in the active site cleft, which identified a potential acetate binding site. The hypothesis that this was a binding site was further supported by alignment of all acetate kinase sequences available from databases, which showed strict conservation of all four residues, and the recent crystal structure of the M. thermophila enzyme with acetate bound in this pocket. Replacement of each residue in the pocket produced variants with Km values for acetate that were 7- to 26-fold greater than that of the wild type, and perturbations of this binding pocket also altered the specificity for longer-chain carboxylic acids and acetyl phosphate. The kinetic analyses of variants combined with structural modeling indicated that the pocket has roles in binding the methyl group of acetate, influencing substrate specificity, and orienting the carboxyl group. The kinetic analyses also indicated that binding of acetyl phosphate is more dependent on interactions of the phosphate group with an unidentified residue than on interactions between the methyl group and the hydrophobic pocket. The analyses also indicated that Phe179 is essential for catalysis, possibly for domain closure. Alignments of acetate kinase, propionate kinase, and butyrate kinase sequences obtained from databases suggested that these enzymes have similar catalytic mechanisms and carboxylic acid substrate binding sites. PMID:15774882

  7. Amphiphilic Membranes

    E-print Network

    Luca Peliti

    2005-07-26

    Contents: 1. Introduction 2. Amphiphilic molecules and the phases they form 3. Isolated membranes: the Helfrich hamiltonian 4. Vesicle shapes 5. Shape fluctuations in vesicles 6. Interacting fluid membranes 7. Conclusions A. Differential equations for vesicle shapes B. The Faddeev-Popov determinant C. One-loop calculation of the renormalization group D. The Liouville model

  8. [Control of nanoparticles in food and biological objects. Report 2. Filtration, centrifugation, spectral methods and electrophoresis].

    PubMed

    Raspopov, R V; Gmoshinski?, I V; Popov, K I; Rykhtik, O V; Khotimchenko, S A

    2012-01-01

    The large number of the analysis methods of engineered nanoparticles and nanoobjects as a part of disperse systems on the basis of principles of a membrane filtration (micro, ultra- and a nanofiltration) ultracentrifugation, spectral methods, including dynamic and static laser light scattering, Raman light scattering, low-angle X-ray scattering, x-ray techniques, laser decomposition spectroscopy, and other methods are developed. Mass spectrometry with inductively coupled plasma can be successfully used in studying of nanomaterials chemical composition in conditions when there is additional independent information on presence of analyzed substance in a nanoscale form. Methods of electrophoresis in a supportive environment and capillary electrophoresis are beginning to be successfully applied in the study of artificial nanomaterials. However, in terms of the identification of engineered nanoparticles and nanoobjects in complex, multicomponent, heterophase systems, that the objects of the environment and, in particular, food products are, all these methods currently can't compete transmission electron microscopy and atomic force microscopy, specified for purpose of certain particular applications, features of which been described in a previous eport in detail. PMID:22888665

  9. 21 CFR 582.5892 - a-Tocopherol acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or Dietary Supplements 1 § 582.5892 a-Tocopherol acetate. (a) Product. a-Tocopherol acetate. (b) Conditions of use....

  10. 21 CFR 582.5892 - a-Tocopherol acetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or Dietary Supplements 1 § 582.5892 a -Tocopherol acetate. (a) Product. a -Tocopherol acetate. (b) Conditions...

  11. 21 CFR 582.5892 - a-Tocopherol acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or Dietary Supplements 1 § 582.5892 a -Tocopherol acetate. (a) Product. a -Tocopherol acetate. (b) Conditions...

  12. 21 CFR 582.5892 - a-Tocopherol acetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or Dietary Supplements 1 § 582.5892 a -Tocopherol acetate. (a) Product. a -Tocopherol acetate. (b) Conditions...

  13. [Epiretinal membranes].

    PubMed

    Dupas, B; Tadayoni, R; Gaudric, A

    2015-11-01

    Idiopathic epiretinal membranes represent a common condition, and are present in approximately 10% of people over the age of 70 years. They are idiopathic in 80% of cases, or may be secondary to various conditions such as a prior retinal detachment, or vascular or inflammatory retinal diseases. The main symptoms are visual loss and metamorphopsia. The diagnosis of epiretinal membrane is currently facilitated by OCT, which provides prognostic and therapeutic decision-making assistance. Surgery for epiretinal membranes is currently well codified through sutureless vitrectomy and dyes. Dissection of the membrane (with or without associated peeling of the internal limiting membrane) ensures good anatomical and functional results, while being relatively minimally invasive. PMID:26454533

  14. Evaluation of cytogenetic and DNA damage caused by thallium(I) acetate in human blood cells.

    PubMed

    Rodríguez-Mercado, Juan J; Hernández-de la Cruz, Heriberto; Felipe-Reyes, Miriam; Jaramillo-Cruz, Eduardo; Altamirano-Lozano, Mario A

    2015-05-01

    Although thallium is detrimental to all living organisms, information regarding the mutagenic and genotoxic effects of this element and its compounds remains scarce. Therefore, we tested the genotoxic and cytotoxic effects of thallium(I) acetate on human peripheral blood cells in vitro using structural chromosomal aberrations (SCAs), sister chromatid exchanges (SCEs), and single-cell gel electrophoresis (at pH >13 or 12.1) analysis. Whole blood samples were incubated with 0.5, 1, 5, 10, 50, or 100 µg/mL thallium salt. Exposure to this metal compound resulted in a clear dose-dependent reduction in the mitotic and replicative indices. An increase in SCAs was evident in the treated group compared with the control group, and significant differences were observed in the percentage of cells with SCAs when metaphase cells were treated with 0.5-10 µg/mL of thallium(I). The SCE test did not reveal any significant differences. We observed that a 1-h treatment with thallium(I) at pH?>?13 significantly increased the comet length for all the concentrations tested; however, at pH 12.1, only the two highest concentrations affected the comet length. These results suggested that thallium(I) acetate induces cytotoxic, cytostatic, and clastogenic effects, as well as DNA damage. PMID:24318865

  15. Expression of acetate permease-like (apl) genes in subsurface communities of Geobacter species under fluctuating acetate concentrations

    SciTech Connect

    Elifantz, H.; N'Guessan, L.A.; Mouser, P.J.; Williams, K H.; Wilkins, M J.; Risso, C.; Holmes, D.E.; Long, P.E.; Lovley, D.R.

    2010-03-01

    The addition of acetate to uranium-contaminated aquifers in order to stimulate the growth and activity of Geobacter species that reduce uranium is a promising in situ bioremediation option. Optimizing this bioremediation strategy requires that sufficient acetate be added to promote Geobacter species growth. We hypothesized that under acetate-limiting conditions, subsurface Geobacter species would increase the expression of either putative acetate symporters genes (aplI and aplII). Acetate was added to a uranium-contaminated aquifer (Rifle, CO) in two continuous amendments separated by 5 days of groundwater flush to create changing acetate concentrations. While the expression of aplI in monitoring well D04 (high acetate) weakly correlated with the acetate concentration over time, the transcript levels for this gene were relatively constant in well D08 (low acetate). At the lowest acetate concentrations during the groundwater flush, the transcript levels of aplII were the highest. The expression of aplII decreased 2-10-fold upon acetate reintroduction. However, the overall instability of acetate concentrations throughout the experiment could not support a robust conclusion regarding the role of apl genes in response to acetate limitation under field conditions, in contrast to previous chemostat studies, suggesting that the function of a microbial community cannot be inferred based on lab experiments alone.

  16. Expression of Acetate Permease-like (apl) Genes in Subsurface Communities of Geobacter Species Under Fluctuating Acetate Concentrations

    SciTech Connect

    Elifantz, H.; N'Guessan, A. L.; Mouser, Paula; Williams, Kenneth H.; Wilkins, Michael J.; Risso, Carla; Holmes, Dawn; Long, Philip E.; Lovley, Derek R.

    2010-09-01

    The addition of acetate to uranium-contaminated aquifers in order to stimulate the growth and activity of Geobacter species that reduce uranium is a promising in situ bioremediation option. Optimizing this bioremediation strategy requires that suf?cient acetate be added to promote Geobacter species growth. We hypothesized that under acetate-limiting conditions, subsurface Geobacter species would increase the expression of either putative acetate symporters genes (aplI and aplII). Acetate was added to a uranium-contaminated aquifer (Ri?e, CO) in two continuous amendments separated by 5 days of groundwater ?ush to create changing acetate concentrations. While the expression of aplI in monitoring well D04 (high acetate) weakly correlated with the acetate concentration over time, the transcript levels for this gene were relatively constant in well D08 (low acetate). At the lowest acetate concentrations during the groundwater ?ush, the transcript levels of aplII were the highest. The expression of aplII decreased 2–10-fold upon acetate reintroduction. However, the overall instability of acetate concentrations throughout the experiment could not support a robust conclusion regarding the role of apl genes in response to acetate limitation under ?eld conditions, in contrast to previous chemostat studies, suggesting that the function of a microbial community cannot be inferred based on lab experiments alone.

  17. DNA sequencing using fluorescence background electroblotting membrane

    DOEpatents

    Caldwell, Karin D. (Salt Lake City, UT); Chu, Tun-Jen (Salt Lake City, UT); Pitt, William G. (Orem, UT)

    1992-01-01

    A method for the multiplex sequencing on DNA is disclosed which comprises the electroblotting or specific base terminated DNA fragments, which have been resolved by gel electrophoresis, onto the surface of a neutral non-aromatic polymeric microporous membrane exhibiting low background fluorescence which has been surface modified to contain amino groups. Polypropylene membranes are preferably and the introduction of amino groups is accomplished by subjecting the membrane to radio or microwave frequency plasma discharge in the presence of an aminating agent, preferably ammonia. The membrane, containing physically adsorbed DNA fragments on its surface after the electroblotting, is then treated with crosslinking means such as UV radiation or a glutaraldehyde spray to chemically bind the DNA fragments to the membrane through said smino groups contained on the surface thereof. The DNA fragments chemically bound to the membrane are subjected to hybridization probing with a tagged probe specific to the sequence of the DNA fragments. The tagging may be by either fluorophores or radioisotopes. The tagged probes hybridized to said target DNA fragments are detected and read by laser induced fluorescence detection or autoradiograms. The use of aminated low fluorescent background membranes allows the use of fluorescent detection and reading even when the available amount of DNA to be sequenced is small. The DNA bound to the membrances may be reprobed numerous times.

  18. Viscometric study of chitosan solutions in acetic acid/sodium acetate and acetic acid/sodium chloride.

    PubMed

    Costa, Cristiane N; Teixeira, Viviane G; Delpech, Marcia C; Souza, Josefa Virginia S; Costa, Marcos A S

    2015-11-20

    A viscometric study was carried out at 25°C to assess the physical-chemical behavior in solution and the mean viscometric molar mass (M¯v) of chitosan solutions with different deacetylation degrees, in two solvent mixtures: medium 1-acetic acid 0.3mol/L and sodium acetate 0.2mol/L; and medium 2-acetic acid 0.1mol/L and sodium chloride 0.2mol/L. Different equations were employed, by graphical extrapolation, to calculate the intrinsic viscosities [?] and the viscometric constants, to reveal the solvent's quality: Huggins (H), Kraemer (K) and Schulz-Blaschke (SB). For single-point determination, the equations used were SB, Solomon-Ciuta (SC) and Deb-Chanterjee (DC), resulting in a faster form of analysis. The values of ?M¯v were calculated by applying the equation of Mark-Houwink-Sakurada. The SB and SC equations were most suitable for single-point determination of [?] and ?M¯v and the Schulz-Blachke constant (kSB), equal to 0.28, already utilized for various systems, can also be employed to analyze chitosan solutions under the conditions studied. PMID:26344278

  19. Characterization of the microdialysis junction interface for capillary electrophoresis/microelectrospray ionization mass spectrometry

    SciTech Connect

    Severs, J.C.; Smith, R.D.

    1997-06-01

    A capillary electrophoresis/electrospray ionization mass spectrometry (CE/ESI-MS) interface, based on an electric circuit across a microdialysis membrane surrounding a short capillary segment closely connected to the separation capillary terminus, is demonstrated to be sensitive, efficient, and rugged. A microspray type ionization emitter produces a stable electrospray at the low flow rates provided by CE and thus avoids both the need for a makeup liquid flow provided by liquid junction or sheath flow interfaces and the subsequent dilution and reduction in sensitivity. Reproducibility studies and comparisons with CE/UV and the CE/sheath flow interface with ESI-MS are presented. Additionally, postrun acidification via the microdialysis junction interface is demonstrated and shown to be capable of denaturing the holomyoglobin protein noncovalent complex while maintaining separation efficiency. 21 refs., 7 figs., 1 tab.

  20. 21 CFR 522.2478 - Trenbolone acetate and estradiol benzoate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Trenbolone acetate and estradiol benzoate. 522... ANIMAL DRUGS § 522.2478 Trenbolone acetate and estradiol benzoate. (a) Specifications. Each implant dose... estradiol benzoate. (2) 4 pellets, each pellet containing 25 mg trenbolone acetate and 3.5 mg...

  1. Kinetics of Ethyl Acetate Synthesis Catalyzed by Acidic Resins

    ERIC Educational Resources Information Center

    Antunes, Bruno M.; Cardoso, Simao P.; Silva, Carlos M.; Portugal, Ines

    2011-01-01

    A low-cost experiment to carry out the second-order reversible reaction of acetic acid esterification with ethanol to produce ethyl acetate is presented to illustrate concepts of kinetics and reactor modeling. The reaction is performed in a batch reactor, and the acetic acid concentration is measured by acid-base titration versus time. The…

  2. Acetate concentrations and oxidation in salt marsh sediments

    NASA Technical Reports Server (NTRS)

    1992-01-01

    Acetate concentrations and rates of acetate oxidation and sulfate reduction were measured in S. alterniflora sediments in New Hampshire and Massachusetts. Pore water extracted from cores by squeezing or centrifugation contained in greater than 0.1 mM acetate and, in some instances, greater than 1.0 mM. Pore water sampled nondestructively contained much less acetate, often less than 0.01 mM. Acetate was associated with roots, and concentrations varied with changes in plant physiology. Acetate turnover was very low whether whole core or slurry incubations were used. Radiotracers injected directly into soils yielded rates of sulfate reduction and acetate oxidation not significantly different from core incubation techniques. Regardless of incubation method, acetate oxidation did not account for a substantial percentage of sulfate reduction. These results differ markedly from data for unvegetated coastal sediments where acetate levels are low, oxidation rate constants are high, and acetate oxication rates greatly exceed rates of sulfate reduction. The discrepancy between rates of acetate oxidation and sulfate reduction in these marsh soils may be due either to the utilization of substrates other than acetate by sulfate reducers or artifacts associated with measurements of organic utilization by rhizosphere bacteria. Care must be taken when interpreting data from salt marsh sediments since the release of material from roots during coring may affect the concentrations of certain compounds as well as influencing results obtained when sediment incubations are employed.

  3. 21 CFR 522.2477 - Trenbolone acetate and estradiol.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...trenbolone acetate and 24 mg estradiol (one implant consisting of 6 pellets, each pellet...trenbolone acetate and 4 mg estradiol) per implant dose. (B) 120 mg trenbolone acetate and 24 mg estradiol (one implant consisting of 7 pellets, each of...

  4. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Vitamin A acetate. 582.5933 Section 582.5933 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of use....

  5. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Vitamin A acetate. 582.5933 Section 582.5933 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of use....

  6. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Vitamin A acetate. 582.5933 Section 582.5933 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of use....

  7. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Vitamin A acetate. 582.5933 Section 582.5933 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of use....

  8. 21 CFR 582.5933 - Vitamin A acetate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Vitamin A acetate. 582.5933 Section 582.5933 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5933 Vitamin A acetate. (a) Product. Vitamin A acetate. (b) Conditions of use....

  9. Perylenetetracarboxylic diimide (PTCDI) nanowires for sensing ethyl acetate in wine.

    PubMed

    Khopkar, Yashdeep; Kojtari, Arben; Swearer, Dayne; Zivanovic, Sandra; Ji, Hai-Feng

    2014-09-01

    We report the application of perylenetetracarboxylic diimide (PTCDI) nanowires for sensing ethyl acetate. The conductivity of the crystalline nano/microwires increases quickly and selectively in the presence of ethyl acetate vapor, but not with water, acid and alcohol vapors, suggesting that the nanowires of PTCDI may be used for monitoring ethyl acetate during a wine manufacturing process. PMID:25924331

  10. 21 CFR 584.200 - Ethyl alcohol containing ethyl acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Ethyl alcohol containing ethyl acetate. The feed additive ethyl alcohol containing ethyl acetate meets the requirement of 27 CFR 21.62, being not less than 92.5 percent ethyl alcohol, each 100 gallons... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ethyl alcohol containing ethyl acetate....

  11. 21 CFR 522.2478 - Trenbolone acetate and estradiol benzoate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Trenbolone acetate and estradiol benzoate. 522... ANIMAL DRUGS § 522.2478 Trenbolone acetate and estradiol benzoate. (a) Specifications. Each implant dose consists of: (1) 8 pellets, each pellet containing 25 milligrams (mg) trenbolone acetate and 3.5...

  12. Absorbance detector for capillary electrophoresis based on light-emitting diodes and photodiodes for the deep-ultraviolet range.

    PubMed

    Bui, Duy Anh; Hauser, Peter C

    2015-11-20

    A new absorbance detector for capillary electrophoresis featuring relatively high intensity light-emitting diodes as radiation sources and photodiodes for the deep-UV range was developed. The direct relationship of absorbance values and concentrations was obtained by emulating Lambert-Beer's law with the application of a beam splitter to obtain a reference signal and a log-ratio amplifier circuitry. The performance of the cell was investigated at 255nm with the detection of sulfanilic, 4-nitrobenzoic, 4-hydroxybenzoic and 4-aminobenzoic acid and the indirect detection of acetate, propionate, butyrate and caproate using benzoate as the displacement dye molecule. Vanillic acid, L-tyrosine and DL-tryptophan as well as the sulfonamides sulfamerazine, sulfathiazole and sulfamethazine were determined at 280nm. Good linearities over 3 orders of magnitude were obtained. The noise level recorded was as low as 50?AU and the drift typically <200?AU/5min. PMID:26091783

  13. EVALUATION OF MEMBRANE PERFORMANCE AND FOULING BY PYROLYSIS-GC/MS

    EPA Science Inventory

    Pyrolysis-GC/MS is used to evaluate the organic foulants found on two types of membranes for three natural waters. olyamide and cellulose acetate membranes are used. aters from Manatee Lake, Harsha Lake, and the Ohio River are used as feed waters. he pyrolysis fragments are class...

  14. Isothermal decomposition of ?-irradiated erbium acetate

    NASA Astrophysics Data System (ADS)

    Mahfouz, R. M.; Al-Shehri, S. M.; Monshi, M. A. S.; Alhaizan, A. I.; El-Salam, N. M. Abd

    Isothermal decomposition of un-irradiated and pre-?-irradiated erbium acetate has been investigated at different temperatures between 583 and 603 K. Irradiation was observed to enhance the rate of decomposition without modifying the mechanism of the thermal decomposition. Thermal decomposition of erbium acetate has been shown to proceed by a nucleation and growth mechanism (Erofe'ev model) both for un-irradiated and pre-?-irradiated samples. The enhancement of the decomposition was found to increase with an increase in the ?-ray dose applied to the sample and may be attributed to an increase in point defects and formation of additional nucleation centers generated in the host lattice. Thermodynamic values of the main decomposition process were calculated and evaluated.

  15. Isothermal decomposition of ?-irradiated samarium acetate

    NASA Astrophysics Data System (ADS)

    Mahfouz, R. M.; Monshi, M. A. S.; Alshehri, S. M.; Abd El-Salam, N. M.

    2000-10-01

    Isothermal decomposition of un-irradiated and pre-?-irradiated samarium acetate has been investigated at different temperatures between 613 and 633 K. Irradiation was observed to enhance the rate of decomposition without modifying the mechanism of thermal decomposition. Thermal decomposition of samarium acetate has been shown to proceed by two-dimensional phase-boundary reaction both for un-irradiated and pre-?-irradiated samples. The enhancement of the decomposition was found to increase with an increase in the ?-ray dose applied to the sample and may be attributed to an increase in point defects and formation of additional nucleation centers generated in the host lattice. Thermodynamic values of the main decomposition process were calculated and evaluated.

  16. Crystalline Membranes

    NASA Technical Reports Server (NTRS)

    Tsapatsis, Michael (Inventor); Lai, Zhiping (Inventor)

    2008-01-01

    In certain aspects, the invention features methods for forming crystalline membranes (e.g., a membrane of a framework material, such as a zeolite) by inducing secondary growth in a layer of oriented seed crystals. The rate of growth of the seed crystals in the plane of the substrate is controlled to be comparable to the rate of growth out of the plane. As a result, a crystalline membrane can form a substantially continuous layer including grains of uniform crystallographic orientation that extend through the depth of the layer.

  17. Multiple-anion nonvolatile acetal (MANA) resists

    NASA Astrophysics Data System (ADS)

    Guevremont, Jeffrey M.; Brainard, Robert L.; Reeves, Scott D.; Zhou, Xin; Nguyen, Thinh B.; Mackevich, Joseph F.; Anderson, Erik H.; Taylor, Gary N.

    2001-08-01

    New acetal or ketal blocking reagents were investigated for use in e-beam lithography and compared with the performance of ethyl vinyl either (EVE). Three blocking groups, (alpha) -Angelicalactone (AL), 6-methylene-5,6-benzo-1,4- dioxane (MBD), and MANA50 (an undisclosed blocking group used to show the potential of this chemistry) were reacted with poly(p-hydroxystyrene) (PHS) under acid catalyzed conditions to form AL-PHS, MBD-PHS, MANA50-PHS. The performance objectives pursued in the design of these new materials was to use acetal (ketal) chemistry to deliver wide process latitudes (e.g. good PED performance and minimal PEB sensitivity), use high molecular weight blocking groups to eliminate outgassing, and use the novel concept of multiple anions to deliver lithographic performance. These new materials are called Multiple Anion Nonvolatile Acetal (MANA) resists. Resists films were exposed with 50kV electrons, post exposure baked (PEB), and developed with 0.26 N TMAH. Resists prepared with the third blocking group, MANA50, gave contrast and imaging performance independent of PEB humidity and were relatively insensitive to PEB temperature and post exposure delay (PED). These resists gave the best resolution (90 nm) and profiles of all the materials tested, as well as showing no outgassing (as measured by film thickness loss).

  18. Recent Developments in Instrumentation for Capillary Electrophoresis and Microchip-Capillary Electrophoresis

    PubMed Central

    Felhofer, Jessica L.; Blanes, Lucas; Garcia, Carlos D.

    2010-01-01

    Over the last years there has been an explosion in the number of developments and applications of capillary electrophoresis (CE) and microchip-CE. In part, this growth has been the direct consequence of recent developments in instrumentation associated with CE. This review, which is focused on contributions published in the last five years, is intended to complement the papers presented in this special issue dedicated to Instrumentation and to provide an overview on the general trend and some of the most remarkable developments published in the areas of high voltage power supplies, detectors, auxiliary components, and compact systems. It also includes few examples of alternative uses of and modifications to traditional CE instruments. PMID:20665910

  19. Biological membranes

    PubMed Central

    Watson, Helen

    2015-01-01

    Biological membranes allow life as we know it to exist. They form cells and enable separation between the inside and outside of an organism, controlling by means of their selective permeability which substances enter and leave. By allowing gradients of ions to be created across them, membranes also enable living organisms to generate energy. In addition, they control the flow of messages between cells by sending, receiving and processing information in the form of chemical and electrical signals. This essay summarizes the structure and function of membranes and the proteins within them, and describes their role in trafficking and transport, and their involvement in health and disease. Techniques for studying membranes are also discussed. PMID:26504250

  20. Nonlinear electrophoresis for purification of soil DNA for metagenomics.

    PubMed

    Engel, Katja; Pinnell, Lee; Cheng, Jiujun; Charles, Trevor C; Neufeld, Josh D

    2012-01-01

    Purification of microbial DNA from soil is challenging due to the co-extraction of humic acids and associated phenolic compounds that inhibit subsequent cloning, amplification or sequencing. Removal of these contaminants is critical for the success of metagenomic library construction and high-throughput sequencing of extracted DNA. Using three different composite soil samples, we compared a novel DNA purification technique using nonlinear electrophoresis on the synchronous coefficient of drag alteration (SCODA) instrument with alternate purification methods such as direct current (DC) agarose gel electrophoresis followed by gel filtration or anion exchange chromatography, Wizard DNA Clean-Up System, and the PowerSoil DNA Isolation kit. Both nonlinear and DC electrophoresis were effective at retrieving high-molecular weight DNA with high purity, suitable for construction of large-insert libraries. The PowerSoil DNA Isolation kit and the nonlinear electrophoresis had high recovery of high purity DNA suitable for sequencing purposes. All methods demonstrated high consistency in the bacterial community profiles generated from the DNA extracts. Nonlinear electrophoresis using the SCODA instrument was the ideal methodology for the preparation of soil DNA samples suitable for both high-throughput sequencing and large-insert cloning applications. PMID:22056233

  1. Capillary electrophoresis with contactless conductivity detection for the determination of carnitine and acylcarnitines in clinical samples.

    PubMed

    Pormsila, Worapan; Morand, Réjane; Krähenbühl, Stephan; Hauser, Peter C

    2011-04-15

    A capillary electrophoresis method with contactless conductivity detection was developed for the quantification of carnitine and six acylcarnitines in plasma and urine samples. The running buffer employed consisted of 500 mmol/L acetic acid, 1.0 mmol/L hydroxypropyl-?-cyclodextrin and 0.05% Tween at a pH of 2.6. Under these conditions, the isomeric valproyl- and octanoyl-carnitines could be distinguished. The linearity was in the range from 5.0 to 200.0 ?mol/L with correlation coefficients between 0.9992 and 0.9997. The limits of detection were between 1.0 and 3.2 ?mol/L. Intra- and inter-day precisions as %RSD were better than 10%. The method allows for direct determination without derivatisation or extraction processes. The method was applied for the quantification of carnitine and acetylcarnitine in plasma pre- and post-exercise, and to measure valproylcarnitine in plasma and urine of patients undergoing valproate therapy. PMID:21435958

  2. Centimeter-scale characterization of biogeochemical gradients at a wetland-aquifer interface using capillary electrophoresis

    USGS Publications Warehouse

    Baez-Cazull, S.; McGuire, J.T.; Cozzarelli, I.M.; Raymond, A.; Welsh, L.

    2007-01-01

    Steep biogeochemical gradients were measured at mixing interfaces in a wetland-aquifer system impacted by landfill leachate in Norman, Oklahoma. The system lies within a reworked alluvial plain and is characterized by layered low hydraulic conductivity wetland sediments interbedded with sandy aquifer material. Using cm-scale passive diffusion samplers, "peepers", water samples were collected in a depth profile to span interfaces between surface water and a sequence of deeper sedimentary layers. Geochemical indicators including electron acceptors, low-molecular-weight organic acids, base cations, and NH4+ were analyzed by capillary electrophoresis (CE) and field techniques to maximize the small sample volumes available from the centimeter-scale peepers. Steep concentration gradients of biogeochemical indicators were observed at various interfaces including those created at sedimentary boundaries and boundaries created by heterogeneities in organic C and available electron acceptors. At the sediment-water interface, chemical profiles with depth suggest that SO42 - and Fe reduction dominate driven by inputs of organic C from the wetland and availability of electron acceptors. Deeper in the sediments (not associated with a lithologic boundary), a steep gradient of organic acids (acetate maximum 8.8 mM) and NH4+ (maximum 36 mM) is observed due to a localized source of organic matter coupled with the lack of electron acceptor inputs. These findings highlight the importance of quantifying the redox reactions occurring in small interface zones and assessing their role on biogeochemical cycling at the system scale. ?? 2007 Elsevier Ltd. All rights reserved.

  3. Determination of biogenic amines in beer and wine by capillary electrophoresis-tandem mass spectrometry.

    PubMed

    Daniel, Daniela; Dos Santos, Vagner Bezerra; Vidal, Denis Tadeu Rajh; do Lago, Claudimir Lucio

    2015-10-16

    A capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) method for the simultaneous assessment of nine biogenic amines (spermine, spermidine, putrescine, cadaverine, histamine, phenylethylamine, tryptamine, tyramine, and urocanic acid) in commercial samples of beer and wine is introduced. The samples were submitted to a simple clean-up step with poly(vinylpolypyrrolidone) followed by filtration. Electrophoretic separation in a polyvinyl alcohol (PVA)-coated capillary using 0.5molL(-1) acetic acid (pH 2.5) as background electrolyte and detection by electrospray-tandem mass spectrometry was employed. The range of the correlation coefficients of the calibration curves of the analyzed compounds was 0.996-0.999, and the limits of detection and limits of quantification were in the range of 1-2?gL(-1) and 3-8?gL(-1), respectively. The recovery values for samples spiked at three concentration levels (0.2, 0.5, and 1.0mgL(-1)) ranged from 87 to 113% with standard deviation not greater than 5.8%. The use of a PVA-coated silica capillary allows suppressing the electroosmotic flow and, consequently, increasing of the separation efficiency. The method was successfully used to determine biogenic amines in commercial samples of beer and wine. PMID:26362807

  4. Determination of enkephalin peptides by nonaqueous capillary electrophoresis with electrochemical detection.

    PubMed

    Psurek, Arndt; Matysik, Frank-Michael; Scriba, Gerhard K E

    2006-03-01

    Nonaqueous capillary electrophoresis with electrochemical detection (NACE-ED) was applied to the analysis of enkephalin peptides. The effect of different buffer compositions on the electrophoretic behavior of methionine enkephalin, leucine enkephalin, and [D-Ala2]-leucine enkephalin was studied. Separation of the protonated and the deprotonated peptides was obtained using ACN/methanol-based electrolyte systems. The electrochemical behavior of the enkephalins was studied by the capillary batch injection analysis technique. NACE-ED yielded well-defined signals in the oxidation mode only for the negatively charged analytes. The optimized BGE for the counterelectroosmotic separation consisted of 10 mM ammonium acetate in ACN/methanol (3:1 v/v). Using a platinum microdisk electrode set to an actual potential of +0.65 V detection limits in the submicromolar range were observed which are about one order of magnitude lower compared to UV detection. Problems concerning EOF instability and electrode fouling caused by water and other neutral sample impurities transported by the EOF can be avoided in the EOF-inverted mode using poly(ethylene glycol)-coated capillaries and an actual working electrode potential of +1.0 V. For the quantification of the enkephalins [D-Ala2]leucine enkephalin was used as internal standard. The practical utility for the determination of enkephalins in spiked plasma samples after SPE was demonstrated. PMID:16523458

  5. Quantitative Determination of Lercanidipine Enantiomers in Commercial Formulations by Capillary Electrophoresis

    PubMed Central

    Lourenço, Luciana Pereira; Aguiar, Fernando Armani; de Oliveira, Anderson Rodrigo Moraes; de Gaitani, Cristiane Masetto

    2015-01-01

    An enantioselective method based on capillary electrophoresis (CE) using cyclodextrin (CD) as chiral selector was developed and validated for determination of lercanidipine (LER) enantiomers, a drug calcium channel blocker which exerts antihypertensive effects of long duration, in a pharmaceutical formulation. Optimum separation of LER enantiomers was obtained on a 50?cm × 50??m id capillary using a sodium acetate buffer solution 200?mmol/L pH 4.0 containing 10?mmol/L of 2,3,6-o-methyl-?-cyclodextrin (TM-?-CD) as background electrolyte. The capillary temperature and voltage were 15°C and 25?kV, respectively, hydrodynamic injection and detection at 237?nm. Linearity was obtained in the range 12.5–100??g/mL for both enantiomers (r ? 0.995). The RSD (%) and relative errors (E, %) obtained in precision and accuracy studies (intraday and interday) were lower than 5%. After validation, the method was applied to quantify the enantiomers of LER in commercial tablets and the results were satisfactory in terms of accuracy and precision, both less than 5%. Therefore, this method was found to be appropriate for enantioselective quality control of LER enantiomers in pharmaceutical formulations. PMID:25821632

  6. Analytical characterization of beet root vacuole membrane

    SciTech Connect

    Marty, F.; Branton, D.

    1980-10-01

    Vacuoles from beet root (Beta vulgaris L. var. esculenta Gurke) isolated by a mechanical procedure were osmotically lysed to separate the membrane and sap components for analysis. Approximately 62% of the vacuole proteins, 70% of the nondialyzable carbohydrates and almost all of the phospholipids and sterols were recovered in the membrane fraction. The vacuole membrane had a phospholipid:protein ratio of 0.68 and a sterol:phospholipid ratio of 0.21. Seventeen complex polar lipids including phosphatides ad glycolipids have been tentatively identified. Phosphatidylcholine (54%) and phosphatidylethanolamine (24%) were the most prominant phosphoglycerides besides phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and phosphatidic acid (1, 4, 5, and 12%, respectively. A putative sulfoglycoside and two major ceramide glycoside-like lipids, resembling those of animal lysosomes, were identified by thin-layer chromatography. High-resolution SDS-acrylamide gel electrophoresis of the polypeptides from the vacuole revealed 15 major bands with apparent molecular weights ranging from 91,000 to 12,000. Selective elution experiments delineated those polypeptides that were peripheral membrane proteins or sap proteins adsorbed to the membrane, and those that exhibited hydrophobic interaction with the lipid core. Lectin labeling results indicated that most of the polypeptides from the membrane and from the sap were glycoproteins probably of the high-mannose type characteristic of lysosomal enzymes that have undergone several stages of posttranslational modification.

  7. Preparative cell electrophoresis at 1 and 0 gravity

    NASA Technical Reports Server (NTRS)

    Van Oss, C. J.; Bronson, P. M.

    1979-01-01

    It is attempted to show that the use of heavy water (D2O) as starting cushion for the cells combines the advantages of the required density difference, with no lasting biochemical or physiochemical influence on the cells. Phosphate buffers of low ionic strength were prepared in distilled water or heavy water. A vertical starch gel electrophoresis was used to support a cylindrical polystyrene electrophoresis tube used for lymphocyte separations, 25 cm in length and 0.75 cm I.D., prepared from a 10 ml disposable pipet. Erythrocyte separations were carried out in a jacketed rectangular plexiglas chamber. It is pointed out that the described preparative D2O gradient electrophoresis method cannot be readily used for the measurement of electrophoretic mobilities for analytical purposes. However, for the preparative separation of cells with only slightly different electrokinetic properties the method appears promising, simple, and entirely inocuous to the cells.

  8. THERMAL DETECTION OF DNA AND PROTEINS DURING GEL ELECTROPHORESIS

    SciTech Connect

    R. JOHNSTON

    2000-08-01

    This is the final report of a three-year, Laboratory-Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The goal of this project was to try to detect unstained, untagged, unlabeled DNA bands in real-time during gel electrophoresis using simple thermal measurements. The technical and ES&H advantages to this approach could potentially be quite significant, especially given the extreme importance of gel electrophoresis to a wide variety of practical and research fields. The project was unable to demonstrate sufficient thermal sensitivity to detect DNA bands. It is clear that we still do not understand the gel electrophoresis phenomenon very well. The temperature control techniques developed during the course of this project have other useful applications.

  9. Inexpensive and safe DNA gel electrophoresis using household materials.

    PubMed

    Ens, S; Olson, A B; Dudley, C; Ross, N D; Siddiqi, A A; Umoh, K M; Schneegurt, M A

    2012-01-01

    Gel electrophoresis is the single most important molecular biology technique and it is central to life sciences research, but it is often too expensive for the secondary science classroom or homeschoolers. A simple safe low-cost procedure is described here that uses household materials to construct and run DNA gel electrophoresis. Plastic containers are fitted with aluminum foil electrodes and 9-V batteries to run food-grade agar-agar gels using aquarium pH buffers and then stained with gentian violet. This activity was tested in a high school biology classroom with significantly positive responses on postactivity reflective surveys. The electrophoresis activity addresses several Life Science Content Standard C criteria, including aspects of cell biology, genetics, and evolution. It also can be used to teach aspects of motion and force in the physical science classroom. PMID:22615228

  10. Can Palladium Acetate Lose Its "Saltiness"? Catalytic Activities of the Impurities in Palladium Acetate.

    PubMed

    Carole, William A; Bradley, Jonathan; Sarwar, Misbah; Colacot, Thomas J

    2015-11-01

    Commercially available palladium acetate often contains two major impurities, whose presence can impact the overall catalytic efficacy. This systematic study provides a comparison of the differences in catalytic activity of pure palladium acetate, Pd3(OAc)6, with the two impurities: Pd3(OAc)5(NO2) and polymeric [Pd(OAc)2]n in a variety of cross-coupling reactions. The solid state (13)C NMR spectra of all three compounds in conjunction with DFT calculations confirm their reported geometries. PMID:26507318

  11. DNA electrophoresis in dilute polymer solutions: a nonbinary mechanism.

    PubMed

    Ekani-Nkodo, Axel; Tinland, Bernard

    2003-05-01

    The dynamical behavior of the neutral polymer (dextran, M(w)=2 x 10(6)) is investigated during DNA electrophoresis in a dilute solution. Using a fluorescence recovery after photobleaching setup, we measured the velocity of fluorescein-labeled dextran induced by the migration of the DNA. We found that each DNA molecule drags a large number of dextrans with it. We show that DNA-dextran interactions are not only binary but long range and indirect. We conclude that the DNA-dextran complex creates a hydrodynamic field that entrains polymers far from the DNA during electrophoresis. PMID:12786191

  12. Definition of performance specifications for automated Analytical Electrophoresis Facility (AAEF)

    NASA Technical Reports Server (NTRS)

    Brooks, D. E.

    1976-01-01

    In order to provide specifications for the automated Analytical Electrophoresis Facility (AAEF) that would satisfy the broadest variety of demands of a future user community, a survey was carried out of all those people who were identified as having published papers on cell electrophoresis in the past four years. A computer search was conducted of the relevant literature from which a list of 87 investigators was derived and defined as the user community for purposes of the mailing. A questionnaire was developed covering the areas of performance which required definition which was subsequently circulated to the user community. Based on the response to this survey performance specifications were assembled.

  13. Separation of DNA by free flow electrophoresis in space.

    PubMed

    Kobayashi, H; Ishii, N

    2001-10-01

    Free flow electrophoresis of a nematode C. elegans DNA was carried out on the space shuttle flight in International Microgravity Laboratory No. 2 (IML-2)., 1994. We selected the C. elegans DNA as the sample of the space experiment for free flow electrophoresis separation. This worm is a useful animal for the study of development and behavior by genetic analysis, and is a good candidate for a complete DNA sequence analysis because the haploid genome size consists of approximately 100 Mb (megabase) distributed on the six chromosomes, only 1/30 of human genome size. (Recently, the entire genomic DNA (97 Mb) sequences of C. elegans have been completed at the last December, 1998, as the first organism of multicellular system. In addition, many of the genes in C. elegans have extensive similarity to their mammalian counterparts. It may be possible to use the technology of free flow electrophoresis to contribute to the DNA analysis of the eukaryote. In this mission we attempted to make real-time communication between the space shuttle and on the ground during the electrophoresis processing. Because the separation tendency of the sample in space was not predicted so perfectly, the requests to the crews of the space shuttle was needed for selecting the fractions to be separated. According to the three dimensional electropherogram (3DEP) figured out the electrophoresis behavior, we were able to recover those fractions and kept in the deep freezer until landing. This real-time monitoring of the electrophoresis was the first evidence during space electrophoresis experiments which was started at Apollo 14, 1974. Unfortunately, some troubles occurred by contamination of bubbles in space nevertheless the post flight analyses of the fractionated samples were succeeded. The DNAs estimated by the DNA probes were not isolated one but the migration tendencies were differ from each other. It was clear that certain parts of the process in this study are advanced; that is, the precise monitoring of the electrophoresis process in space was accomplished, in real-time, by using 3DEP. In addition, the detection of DNA of recovered samples indicates that all the processes during the study were kept to be free from biological contamination. These results were published in following paper and reviewed in the book. PMID:11799254

  14. Gel Electrophoresis of Gold-DNA Nano-Conjugates

    SciTech Connect

    Pellegrino, T.; Sperling, R.A.; Alivisatos, A.P.; Parak, W.J.

    2006-01-10

    Single stranded DNA of different lengths and different amounts was attached to colloidal phosphine stabilized Au nanoparticles. The resulting conjugates were investigated in detail by a gel electrophoresis study based on 1200 gels. We demonstrate how these experiments help to understand the binding of DNA to Au particles. In particular we compare specific attachment of DNA via gold-thiol bonds with nonspecific adsorption of DNA. The maximum number of DNA molecules that can be bound per particle was determined. We also compare several methods to used gel electrophoresis for investigating the effective diameter of DNA-Au conjugates, such as using a calibration curve of particles with known diameters and Ferguson plots.

  15. EOS production on the Space Station. [Electrophoresis Operations/Space

    NASA Technical Reports Server (NTRS)

    Runge, F. C.; Gleason, M.

    1986-01-01

    The paper discusses a conceptual integration of the equipment for EOS (Electrophoresis Operations/Space) on the Space Station in the early 1990s. Electrophoresis is a fluid-constituent separation technique which uses forces created by an electrical field. Aspects covered include EOS equipment and operations, and Space Station installations involving a pressurized module, a resupply module, utility provisions and umbilicals and crew involvement. Accommodation feasibility is generally established, and interfaces are defined. Space Station production of EOS-derived pharmaceuticals will constitute a significant increase in capability compared to precursor flights on the Shuttle in the 1980s.

  16. Effect of iodination on human growth hormone and prolactin: characterized by bioassay, radioimmunoassay, radioreceptor assay, and electrophoresis

    SciTech Connect

    Hughes, J.P.; Tanaka, T.; Gout, P.W.; Beer, C.T.; Noble, R.L.; Friesen, H.G.

    1982-09-01

    Human GH (hGH) and PRL (hPRL) were iodinated using lactoperoxidase. The iodinated hormones were characterized by RIA, radioreceptor assay (RRA), and bioassay (BA) using the Nb2 Node lymphoma cell line. The proportion of tracer that could bind to rat liver membranes or rabbit antibodies was determined, and the distribution of iodinated hormones was examined using polyacrylamide gel electrophoresis. Excess antibody was capable of precipitating 87.9% of the radioactivity associated with the hGH tracer and 86.0% of the hPRL tracer. The maximal specific binding to a liver membrane preparation averaged 67.3% of the (/sup 125/I)iodo-hGH radioactivity and 48.8% of the (/sup 125/I)iodo-hPRL radioactivity. The respective BA and RRA activity estimates for (/sup 125/)iodo-hGH averaged 90% and 114% of the activity measured by the RIA. For (/sup 125/I)iodo-hPRL, the values were 75% by BA and 68% by RRA. The bioactivity profiles of iodinated hGH and hPRL shifted anodally on polyacrylamide gel electrophoresis in comparison to the bioactivity distribution of the respective uniodinated hormones. Iodine incorporation rather than oxidation appeared to be responsible for the shift. After electrophoresis, all eluates which contained significant radioactivity were active in the BA and RIA. Furthermore, specific activities calculated from the bioactive hormone and radioactivity in each electrophoretic segment agreed well with the average specific activity estimated from the amount of iodine incorporated into the protein peak upon gel filtration. These data suggest that hGH and hPRL to a major degree retain biological integrity after iodination.

  17. Effect of iodination on human growth hormone and prolactin: characterized by bioassay, radioimmunoassay, radioreceptor assay, and electrophoresis

    SciTech Connect

    Hughes, J.P.; Tanaka, T.; Gout, P.W.; Beer, C.T.; Noble, R.L.; Friesen, H.G.

    1982-09-01

    Human GH (hGH) and PRL (hPRL) were iodinated using lactoperoxidase. The iodinated hormones were characterized by RIA, radioreceptor assay (RRA), and bioassay (BA) using the Nb2 Node lymphoma cell line. The proportion of tracer that could bind to rat liver membranes or rabbit antibodies was determined, and the distribution of iodinated hormones was examined using polyacrylamide gel electrophoresis. Excess antibody was capable of precipitating 87.9% of the radioactivity associated with the hGH tracer and 86.0% of the hPRL tracer. The maximal specific binding to a liver membrane preparation averaged 67.3% of the (/sup 125/I)iodo-hGH radioactivity and 48.8% of the (/sup 125/I)iodo-hPRL radioactivity. The respective BA and RRA activity estimates for (/sup 125/I)iodo-hGH averaged 90% and 114% of the activity measured by the RIA. For (/sup 125/I)iodo-hPRL, the values were 75% by BA and 68% by RRA. The bioactivity profiles of iodinated hGH and hPRL shifted anodally on polyacrylamide gel electrophoresis in comparison to the bioactivity distribution of the respective uniodinated hormones. Iodine incorporation rather than oxidation appeared to be responsible for the shift. After electrophoresis, all eluates which contained significant radioactivity were active in the BA and RIA. Furthermore, specific activities calculated from the bioactive hormone and radioactivity in each electrophoretic segment agreed well with the average specific activity estimated from the amount of iodine incorporated into the protein peak upon gel filtration. These data suggest that hGH and hPRL to a major degree retain biological integrity after iodination.

  18. Electron tunneling studies of Mn12-Acetate 

    E-print Network

    Ma, Lianxi

    2008-10-10

    dependence. In the region |V |?0.04 V, we find a zero-bias feature (ZBF) in which the differential conductance is suppressed. In some samples, we observe I?V staircases which we attribute to electrons “hopping” between the electrodes and the molecules.... The observed hystere- sis was attributed to the slow relaxation of molecules re-orienting within the junction. Abrupt conductance jumps at a bias voltage of -0.12 V were also observed and may indicate state transitions in the Mn 12 -Acetate molecules...

  19. [Analysis of the character of film decomposition of starch acetate (SA) coated urea by infrared spectrum].

    PubMed

    Li, Dong-Po; Wu, Zhi-Jie; Liang, Cheng-Hua; Chen, Li-Jun; Zhang, Yu-Lan; Nie, Yan-Xia

    2012-06-01

    The degradability characteristics of film with 4 kinds of starch acetate coated and inhibitors amended urea were analyzed by FTIR, which was purposed to supply theoretical basis for applying starch acetate coated urea fertilizers in farming. The result showed that the chemical component, molecule structure and material form of the membrane were not changed because of adding different inhibitors to urea. The main peaks of the film degradation process were brought by the H--O, --OH, CO2, C==O, --CH2, --CH3, C--O, C--O--H and C--O--C vibrancy in asymmetry and symmetry. In brown soil, the trend of absorbing value of the most high peak was 0>15>30>60>90>120>150>310 d. The infrared spectra of 4 kinds of fertilizers were not different remarkably, and the film was comparatively slowly degraded before 15 d. But a majority of the film had been already degraded after 150 days. The main components of film materials were degraded fastest in 310 days. The speed of film degradation wasn't more impacted by different inhibitors. The characteristic of starch acetate film degradation may be monitored entirely and degradation speed difference of the film could be represented through infrared spectrum. PMID:22870631

  20. Elemental sulfur and acetate can support life of a novel strictly anaerobic haloarchaeon.

    PubMed

    Sorokin, Dimitry Y; Kublanov, Ilya V; Gavrilov, Sergei N; Rojo, David; Roman, Pawel; Golyshin, Peter N; Slepak, Vladlen Z; Smedile, Francesco; Ferrer, Manuel; Messina, Enzo; La Cono, Violetta; Yakimov, Michail M

    2016-01-01

    Archaea domain is comprised of many versatile taxa that often colonize extreme habitats. Here, we report the discovery of strictly anaerobic extremely halophilic euryarchaeon, capable of obtaining energy by dissimilatory reduction of elemental sulfur using acetate as the only electron donor and forming sulfide and CO2 as the only products. This type of respiration has never been observed in hypersaline anoxic habitats and is the first example of such metabolic capability in the entire Archaea domain. We isolated and cultivated these unusual organisms, selecting one representative strain, HSR2, for detailed characterization. Our studies including physiological tests, genome sequencing, gene expression, metabolomics and [(14)C]-bicarbonate assimilation assays revealed that HSR2 oxidized acetate completely via the tricarboxylic acid cycle. Anabolic assimilation of acetate occurred via activated glyoxylate bypass and anaplerotic carboxylation. HSR2 possessed sulfurtransferase and an array of membrane-bound polysulfide reductase genes, all of which were expressed during the growth. Our findings suggest the biogeochemical contribution of haloarchaea in hypersaline anoxic environments must be reconsidered. PMID:25978546

  1. Field-amplified sample stacking capillary electrophoresis with electrochemiluminescence applied to the determination of illicit drugs on banknotes.

    PubMed

    Xu, Yuanhong; Gao, Ying; Wei, Hui; Du, Yan; Wang, Erkang

    2006-05-19

    Capillary electrophoresis (CE) with Ru(bpy)3(2+) electrochemiluminescence (ECL) detection system was established to the determination of contamination of banknotes with controlled drugs and a high efficiency on-column field-amplified sample stacking (FASS) technique was also optimized to increase the ECL intensity. The method was illustrated using heroin and cocaine, which are two typical and popular illicit drugs. Highest sample stacking was obtained when 0.01 mM acetic acid was chosen for sample dissolution with electrokinetical injection for 6 s at 17 kV. Under the optimized conditions: ECL detection at 1.2 V, separation voltage 10.0 kV, 20 mM phosphate-acetate (pH 7.2) as running buffer, 5 mM Ru(bpy)3(2+) with 50 mM phosphate-acetate (pH 7.2) in the detection cell, the standard curves were linear in the range of 7.50x10(-8) to 1.00x10(-5) M for heroin and 2.50x10(-7) to 1.00x10(-4) M for cocaine and detection limits of 50 nM for heroin and 60 nM for cocaine were achieved (S/N = 3), respectively. Relative standard derivations of the ECL intensity and the migration time were 3.50 and 0.51% for heroin and 4.44 and 0.12% for cocaine, respectively. The developed method was successfully applied to the determination of heroin and cocaine on illicit drug contaminated banknotes without any damage of the paper currency. A baseline resolution for heroin and cocaine was achieved within 6 min. PMID:16616928

  2. Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures -A Single Experiment Reveals Equilibrium and Kinetic Parameters of Protein-DNA

    E-print Network

    Krylov, Sergey

    into the capillary and subjected to electrophoresis under nonequilibrium conditions. The new method allowsNonequilibrium Capillary Electrophoresis of Equilibrium Mixtures - A Single Experiment Reveals method, nonequilibrium capillary electrophoresis of equilibrium mixtures (NECEEM), and describe its use

  3. Using Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures for the Determination of

    E-print Network

    Krylov, Sergey

    Using Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures for the Determination as nonequilibrium capillary electrophoresis of equilibrium mixtures (NE- CEEM). Conceptually, a calibration curve apparatus or under otherwise differ- ent conditions (cooling efficiency, length and diameter

  4. Analysis of Bacterial Communities in Seagrass Bed Sediments by Double-Gradient Denaturing Gradient Gel Electrophoresis

    E-print Network

    Sherman, Tim

    -Gradient Denaturing Gradient Gel Electrophoresis of PCR-Amplified 16S rRNA Genes J.B. James1 , T.D. Sherman2 and R denaturing gradient gel electrophoresis (DG- DGGE) was used to generate banding patterns from

  5. Phytogenic biosynthesis and emission of methyl acetate.

    PubMed

    Jardine, Kolby; Wegener, Frederik; Abrell, Leif; van Haren, Joost; Werner, Christiane

    2014-02-01

    Acetylation of plant metabolites fundamentally changes their volatility, solubility and activity as semiochemicals. Here we present a new technique termed dynamic (13) C-pulse chasing to track the fate of C1-3 carbon atoms of pyruvate into the biosynthesis and emission of methyl acetate (MA) and CO2 . (13) C-labelling of MA and CO2 branch emissions respond within minutes to changes in (13) C-positionally labelled pyruvate solutions fed through the transpiration stream. Strong (13) C-labelling of MA emissions occurred only under pyruvate-2-(13) C and pyruvate-2,3-(13) C feeding, but not pyruvate-1-(13) C feeding. In contrast, strong (13) CO2 emissions were only observed under pyruvate-1-(13) C feeding. These results demonstrate that MA (and other volatile and non-volatile metabolites) derive from the C2,3 atoms of pyruvate while the C1 atom undergoes decarboxylation. The latter is a non-mitochondrial source of CO2 in the light generally not considered in studies of CO2 sources and sinks. Within a tropical rainforest mesocosm, we also observed atmospheric concentrations of MA up to 0.6 ppbv that tracked light and temperature conditions. Moreover, signals partially attributed to MA were observed in ambient air within and above a tropical rainforest in the Amazon. Our study highlights the potential importance of acetyl coenzyme A (CoA) biosynthesis as a source of acetate esters and CO2 to the atmosphere. PMID:23862653

  6. Immunotoxicity of trenbolone acetate in Japanese quail

    USGS Publications Warehouse

    Quinn, M.J.; McKernan, M.; Lavoie, E.T.; Ottinger, M.A.

    2007-01-01

    Trenbolone acetate is a synthetic androgen that is currently used as a growth promoter in many meat-exporting countries. Despite industry laboratories classifying trenbolone as nonteratogenic, data showed that embryonic exposure to this androgenic chemical altered development of the immune system in Japanese quail. Trenbolone is lipophilic, persistent, and released into the environment in manure used as soil fertilizer. This is the first study to date to assess this chemical's immunotoxic effects in an avian species. A one-time injection of trenbolone into yolks was administered to mimic maternal deposition, and subsequent effects on the development and function of the immune system were determined in chicks and adults. Development of the bursa of Fabricius, an organ responsible for development of the humoral arm of the immune system, was disrupted, as indicated by lower masse, and smaller and fewer follicles at day 1 of hatch. Morphological differences in the bursas persisted in adults, although no differences in either two measures of immune function were observed. Total numbers of circulating leukocytes were reduced and heterophil-lymphocyte ratios were elevated in chicks but not adults. This study shows that trenbolone acetate is teratogenic and immunotoxic in Japanese quail, and provides evidence that the quail immune system may be fairly resilient to embryonic endocrine-disrupting chemical-induced alterations following no further exposure posthatch.

  7. Proteomic profiling of macrophages by 2D electrophoresis.

    PubMed

    Bouvet, Marion; Turkieh, Annie; Acosta-Martin, Adelina E; Chwastyniak, Maggy; Beseme, Olivia; Amouyel, Philippe; Pinet, Florence

    2014-01-01

    The goal of the two-dimensional (2D) electrophoresis protocol described here is to show how to analyse the phenotype of human cultured macrophages. The key role of macrophages has been shown in various pathological disorders such as inflammatory, immunological, and infectious diseases. In this protocol, we use primary cultures of human monocyte-derived macrophages that can be differentiated into the M1 (pro-inflammatory) or the M2 (anti-inflammatory) phenotype. This in vitro model is reliable for studying the biological activities of M1 and M2 macrophages and also for a proteomic approach. Proteomic techniques are useful for comparing the phenotype and behaviour of M1 and M2 macrophages during host pathogenicity. 2D gel electrophoresis is a powerful proteomic technique for mapping large numbers of proteins or polypeptides simultaneously. We describe the protocol of 2D electrophoresis using fluorescent dyes, named 2D Differential Gel Electrophoresis (DIGE). The M1 and M2 macrophages proteins are labelled with cyanine dyes before separation by isoelectric focusing, according to their isoelectric point in the first dimension, and their molecular mass, in the second dimension. Separated protein or polypeptidic spots are then used to detect differences in protein or polypeptide expression levels. The proteomic approaches described here allows the investigation of the macrophage protein changes associated with various disorders like host pathogenicity or microbial toxins. PMID:25408153

  8. Density gradient electrophoresis of cells in a reversible gel.

    PubMed

    Plank, L D; Kunze, M E; Gaines, R A; Todd, P

    1988-10-01

    Density gradient electrophoresis permits the separation of cell types according to surface charge density with high resolution. Any source of flow compromises the resolving power of density gradient electrophoresis. Although procedures have been devised to successfully counteract electroosmotic and convective flows, the final collection of separands requires that they be pumped out of the electrophoresis column. Experiments were therefore designed to test the hypothesis that this flow could also be eliminated by trapping the separated bands in a gel, from which they could be collected by slicing the gel cylinder. Glutaraldehyde-fixed rat and rabbit erythrocytes were used as test particles in a phosphate-buffered isotonic Ficoll-sucrose density gradient in a 2.2 cm diameter, thermostated vertical glass column that could be opened at both ends. Two types of agarose were used as gel polymers: Electrophoresis grade agarose (J.T. Baker Chemical Co.) at final concentrations of 0.1 to 0.25% and SeaPrep ultralow gelling agarose (Marine Colloids Div., FMC Corp.) at a final concentration of 1.0%. Electrophoretic separability of the test particles and fluid stability were tested independently at 55 degrees C and 32 degrees C at which the two agaroses were, respectively, liquid. The experiments demonstrated that the higher temperatures required and the presence of agarose compromised neither the stability of the density gradient nor the migration properties of the cells, and cells can be separated in a sol at a temperature that is compatible with cell viability. PMID:2468483

  9. An Inexpensive Device for Capillary Electrophoresis with Fluorescence Detection

    ERIC Educational Resources Information Center

    Anderson, Greg; Thompson, Jonathan E.; Shurrush, Khriesto

    2006-01-01

    We describe an inexpensive device for performing capillary electrophoresis (CE) separations with fluorescence detection. As a demonstration of the device's utility we have determined the mass of riboflavin in a commercially available dietary supplement. The device allows for separation of riboflavin in [asymptotically equivalent to] 100 s with a…

  10. Pulsed-field gel electrophoresis typing of Staphylococcus aureus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pulsed-field gel electrophoresis (PFGE) is the most applied and effective genetic typing method for epidemiological studies and investigation of foodborne outbreaks caused by different pathogens, including Staphylococcus aureus. The technique relies on analysis of large DNA fragments generated by th...

  11. How it all began: a personal history of gel electrophoresis.

    PubMed

    Smithies, Oliver

    2012-01-01

    Arne Tiselius' moving boundary electrophoresis method was still in general use in 1951 when this personal history begins, although zonal electrophoresis with a variety of supporting media (e.g., filter paper or starch grains) was beginning to replace it. This chapter is an account of 10 years of experiments carried out by the author during which molecular sieving gel electrophoresis was developed and common genetic variants of two proteins, haptoglobin and transferrin, were discovered in normal individuals. Most of the figures are images of pages from the author's laboratory notebooks, which are still available, so that some of the excitement of the time and the humorous moments are perhaps apparent. Alkaline gels, acidic gels with and without denaturants, vertical gels, two-dimensional gels, and gels with differences in starch concentration are presented. The subtle details that can be discerned in these various gels played an indispensable role in determining the nature of the change in the haptoglobin gene (Hp) that leads to the polymeric series characteristic of Hp ( 2 ) /Hp ( 2 ) homozygotes. Where possible, the names of scientific friends who made this saga of gel electrophoresis so memorable and enjoyable are gratefully included. PMID:22585472

  12. Temperature Effects on Electrophoresis Anita Rogacs and Juan G. Santiago*

    E-print Network

    Santiago, Juan G.

    Temperature Effects on Electrophoresis Anita Rogacs and Juan G. Santiago* Department of Mechanical: We present a model capturing the important contributors to the effects of temperature on the observable electrophoretic mobilities of small ions, and on solution conductivity and pH. Our temperature

  13. Application of capillary electrophoresis in agricultural and soil chemistry research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As a modern analytical technique, capillary electrophoresis (CE) has become an attractive method for characterizing molecules wit high structural complexity and a wide range of molecular weights. CE can be used to analyze many natural chemical components such as acids, biogenic amines, peptides, pro...

  14. Separation of rat pituitary secretory granules by continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Hayes, Daniel; Exton, Carrie; Salada, Thomas; Shellenberger, Kathy; Waddle, Jenny; Hymer, W. C.

    1990-01-01

    The separation of growth hormone-containing cytoplasmic secretory granules from the rat pituitary gland by continuous flow electrophoresis is described. The results are consistent with the hypothesis that granule subpopulations can be separated due to differences in surface charge; these, in turn, may be related to the oligomeric state of the hormone.

  15. Procedure for Mobility Gel Electrophoresis Prepare Linearized Plasmid DNA

    E-print Network

    Turro, Claudia

    Procedure for Mobility Gel Electrophoresis Prepare Linearized Plasmid DNA 1. Purified pUC18 plasmid it is recommended (Quiagen Handbook) that for long plasmids (pUC18 ~ 2600 bps) a 10 x excess be used or 10 Units Sma

  16. SEPARATION OF GLUTEN PROTEINS BY HIGH PERFORMANCE CAPILLARY ELECTROPHORESIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High performance capillary electrophoresis (HPCE) is an analytical method that uses a voltage differential to accurately move solvents and solutes through a capillary. HPCE is a relative newcomer to the field of cereal chemistry, it utilizes small inner diameter capillaries as an anti-convective med...

  17. Solid friction in gel electrophoresis S. F. Burlatskya)

    E-print Network

    Deutch, John

    Solid friction in gel electrophoresis S. F. Burlatskya) and John M. Deutch Department of Chemistry 1995 We study the influence of solid frictional forces acting on polymer chains moving in a random environment. We show that the total reduction in the chain tension resulting from the small friction between

  18. Electrostatic Stabilization of Colloids in Carbon Dioxide: Electrophoresis and Dielectrophoresis

    E-print Network

    Electrostatic Stabilization of Colloids in Carbon Dioxide: Electrophoresis and Dielectrophoresis in supercritical fluid carbon dioxide (scCO2). Herein we demonstrate that colloids may also be stabilized in CO2 the behavior of steric stabilization in compressed supercritical fluids1-3 including carbon dioxide,4

  19. Gel Electrophoresis--The Easy Way for Students

    ERIC Educational Resources Information Center

    VanRooy, Wilhelmina; Sultana, Khalida

    2010-01-01

    This article describes a simple, inexpensive, easy to conduct gel-electrophoresis activity using food dyes. It is an alternative to the more expensive counterparts which require agarose gel, DNA samples, purchased chamber and Tris-borate-EDTA buffer. We suggest some learning activities for senior biology students along with comments on several…

  20. Capillary electrophoresis application in metal speciation and complexation characterization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary electrophoresis is amenable to the separation of metal ionic species and the characterization of metal-ligand interactions. This book chapter reviews and discusses three representative case studies in applications of CE technology in speciation and reactions of metal with organic molecules...

  1. Membrane magic

    SciTech Connect

    Buecker, B.

    2005-09-01

    The Kansas Power and Light Co.'s La Cyne generating station has found success with membrane filtration water pretreatment technology. The article recounts the process followed in late 2004 to install a Pall Aria 4 microfilter in Unit 1 makeup water system at the plant to produce cleaner water for reverse osmosis feed. 2 figs., 2 photos.

  2. Thermal decomposition of acetate: III. Catalysis by mineral surfaces

    NASA Astrophysics Data System (ADS)

    Bell, Julie L. S.; Palmer, Donald A.; Barnes, H. L.; Drummond, S. E.

    1994-10-01

    The kinetics of thermal decarboxylation of aqueous solutions of acetic acid and sodium acetate were evaluated at 335 and 355°C in contact with various surfaces as potential catalysts. Quartz, fused quartz, calcite, natural pyrite, titanium oxide, and Au apparently do not catalyze aqueous decarboxylation reactions, in contrast to Pyrex, Ca-montmorillonite, Fe-bearing montmorillonite, hematite, synthetic pyrite, and magnetite. The dependence of the rate of acetic acid decarboxylation on the surface area of pyrite per unit solution volume was also studied. The results show that the decarboxylation of acetic acid and acetate is catalyzed heterogeneously, with the cleavage of the C-C bond occurring while the acetate molecule is adsorbed onto a surface. Entropies and enthalpies of activation obtained from these experiments are compatible with the isokinetic relationship established previously for acetic acid and acetate under similar experimental conditions, indicating the existence of a common rate-determining step. Experimental evidence indicates that oxidation of acetic acid can occur with hematite and defected magnetite. These oxidative decomposition reactions differ from the decarboxylation reaction in that CO 2 and polycondensates are produced instead of CO 2 and CH 4.

  3. Negative Pressure Wound Therapy of Chronically Infected Wounds Using 1% Acetic Acid Irrigation

    PubMed Central

    Lee, Byeong Ho; Lee, Hye Kyung; Kim, Hyoung Suk; Moon, Min Seon; Suh, In Suck

    2015-01-01

    Background Negative-pressure wound therapy (NPWT) induces angiogenesis and collagen synthesis to promote tissue healing. Although acetic acid soaks normalize alkali wound conditions to raise tissue oxygen saturation and deconstruct the biofilms of chronic wounds, frequent dressing changes are required. Methods Combined use of NPWT and acetic acid irrigation was assessed in the treatment of chronic wounds, instilling acetic acid solution (1%) beneath polyurethane membranes twice daily for three weeks under continuous pressure (125 mm Hg). Clinical photographs, pH levels, cultures, and debrided fragments of wounds were obtained pre- and posttreatment. Tissue immunostaining (CD31, Ki-67, and CD45) and reverse transcription-polymerase chain reaction (vascular endothelial growth factor [VEGF], vascular endothelial growth factor receptor [VEGFR]; procollagen; hypoxia-inducible factor 1 alpha [HIF-1-alpha]; matrix metalloproteinase [MMP]-1,-3,-9; and tissue inhibitor of metalloproteinase [TIMP]) were also performed. Results Wound sizes tended to diminish with the combined therapy, accompanied by drops in wound pH (weakly acidic or neutral) and less evidence of infection. CD31 and Ki-67 immunostaining increased (P<0.05) post-treatment, as did the levels of VEGFR, procollagen, and MMP-1 (P<0.05), whereas the VEGF, HIF-1-alpha, and MMP-9/TIMP levels declined (P<0.05). Conclusions By combining acetic acid irrigation with negative-pressure dressings, both the pH and the size of chronic wounds can be reduced and infections be controlled. This approach may enhance angiogenesis and collagen synthesis in wounds, restoring the extracellular matrix. PMID:25606491

  4. Extracellular protease digestion to evaluate membrane protein cell surface localization.

    PubMed

    Besingi, Richard N; Clark, Patricia L

    2015-12-01

    Membrane proteins have crucial roles in signaling and as anchors for cell surface display. Proper secretion of a membrane protein can be evaluated by its susceptibility to digestion by an extracellular protease, but this requires a crucial control to confirm membrane integrity during digestion. This protocol describes how to use this approach to determine how efficiently a protein is secreted to the outer surface of Gram-negative bacteria. Its success relies upon careful selection of an appropriate intracellular reporter protein that will remain undigested if the membrane barrier remains intact, but that is rapidly digested when cells are lysed before evaluation. Reporter proteins that are resistant to proteases (e.g., maltose-binding protein) do not return accurate results; in contrast, proteins that are more readily digested (e.g., SurA) serve as more sensitive reporters of membrane integrity, yielding more accurate measurements of membrane protein localization. Similar considerations apply when evaluating membrane protein localization in other contexts, including eukaryotic cells and organelle membranes. Evaluating membrane protein localization using this approach requires only standard biochemistry laboratory equipment for cell lysis, gel electrophoresis and western blotting. After expression of the protein of interest, this procedure can be completed in 4 h. PMID:26584447

  5. Water vapor diffusion membrane development

    NASA Technical Reports Server (NTRS)

    Tan, M. K.

    1977-01-01

    An application of the water vapor diffusion technique is examined whereby the permeated water vapor is vented to space vacuum to alleviate on-board waste storage and provide supplemental cooling. The work reported herein deals primarily with the vapor diffusion-heat rejection (VD-HR) as it applies to the Space Shuttle. A stack configuration was selected, designed and fabricated. An asymmetric cellulose acetate membrane, used in reverse osmosis application was selected and a special spacer was designed to enhance mixing and promote mass transfer. A skid-mount unit was assembled from components used in the bench unit although no attempt was made to render it flight-suitable. The operating conditions of the VD-HR were examined and defined and a 60-day continuous test was carried out. The membranes performed very well throughout the test; no membrane rupture and no unusual flux decay was observed. In addition, a tentative design for a flight-suitable VD-HR unit was made.

  6. Oxidation of indole-3-acetic acid to oxindole-3-acetic acid by an enzyme preparation from Zea mays

    NASA Technical Reports Server (NTRS)

    Reinecke, D. M.; Bandurski, R. S.

    1988-01-01

    Indole-3-acetic acid is oxidized to oxindole-3-acetic acid by Zea mays tissue extracts. Shoot, root, and endosperm tissues have enzyme activities of 1 to 10 picomoles per hour per milligram protein. The enzyme is heat labile, is soluble, and requires oxygen for activity. Cofactors of mixed function oxygenase, peroxidase, and intermolecular dioxygenase are not stimulatory to enzymic activity. A heat-stable, detergent-extractable component from corn enhances enzyme activity 6- to 10-fold. This is the first demonstration of the in vitro enzymic oxidation of indole-3-acetic acid to oxindole-3-acetic acid in higher plants.

  7. Role of Hydrogen-Bonding in Nonelectrolyte Diffusion through Dense Artificial Membranes

    PubMed Central

    Gary-Bobo, C. M.; DiPolo, R.; Solomon, A. K.

    1969-01-01

    The diffusion of two series of alcohols and amides through complex cellulose acetate membranes was studied. The thin dense part of these membranes behaves as a nonporous layer of low water content. In this layer, called the skin, the solute diffusion coefficients, ?, depend upon size, steric configuration, and the partition coefficient, K8, between membrane and bathing solution. From the experimental values of ? and K8, the over-all friction, f, experienced by the solutes in the membrane was computed. It was found that f depends upon the chemical nature of the solute and is related to hydrogen-bonding ability. In the coarse, porous layer of the cellulose acetate membrane, diffusion occurs mainly through aqueous channels. In this instance also the hydrogen-bonding ability of the solute seems to exercise a smaller but significant influence. PMID:5806595

  8. Role of hydrogen-bonding in nonelectrolyte diffusion through dense artificial membranes.

    PubMed

    Gary-Bobo, C M; DiPolo, R; Solomon, A K

    1969-09-01

    The diffusion of two series of alcohols and amides through complex cellulose acetate membranes was studied. The thin dense part of these membranes behaves as a nonporous layer of low water content. In this layer, called the skin, the solute diffusion coefficients, omega, depend upon size, steric configuration, and the partition coefficient, K(8), between membrane and bathing solution. From the experimental values of omega and K(8), the over-all friction, f, experienced by the solutes in the membrane was computed. It was found that f depends upon the chemical nature of the solute and is related to hydrogen-bonding ability. In the coarse, porous layer of the cellulose acetate membrane, diffusion occurs mainly through aqueous channels. In this instance also the hydrogen-bonding ability of the solute seems to exercise a smaller but significant influence. PMID:5806595

  9. Supported Lipid Bilayer Electrophoresis: A New Paradigm in Membrane Biophysics and Separations 

    E-print Network

    Pace, Hudson 1982-

    2012-11-28

    The motivation of this work was to produce novel analytical techniques capable of probing the physical properties of the cell surface. Many researchers have used supported lipid bilayers (SLBs) as models to study the structure and function...

  10. Antitumor and apoptotic activities of the chemical constituents from the ethyl acetate extract of Artemisia indica.

    PubMed

    Zeng, Ying-Tong; Jiang, Jian-Min; Lao, Hai-Yan; Guo, Jie-Wen; Lun, Yu-Ning; Yang, Min

    2015-03-01

    Cancer is one of the most eminent diseases of modern times and numerous natural products derived from medicinal plants have been identified as potential sources of antitumor drugs. A successful anticancer drug must target or inhibit tumor cells whilst causing minimal damage to healthy cells. The present study aimed to investigate the antitumor efficacy of ethyl acetate extract, and other isolated compounds from Artemisia indica, on MCF?7, BHY, Miapaca?2, Colo?205 and A?549 cell lines. The apoptotic activity of the compounds was studied using flow cytometry. The different cancer cell lines were treated with the ethyl acetate extract and varying concentrations of compounds (denoted a?g) isolated from the A. indica. The cytotoxicity was evaluated by MTT assay and the apoptotic properties of the compounds and the extract were assessed using flow cytometry. In MCF?7 cells, the effect on mitochondrial membrane potential loss (??m) induced by compounds b and d was also studied. Bioassay?guided fractionation of the ethyl acetate extract from the shoot and root parts of A. indica led to the identification of the compounds a?g as: 5?hydroxy?3,7,4'?trimethoxyflavone; ludartin; maackiain; lupeol; cis?matricaria ester; trans?matricaria ester; and 6?methoxy?7,8?methylenedioxy coumarin, respectively. All the compounds exhibited mild to potent inhibition of cell proliferation in all the cell lines, with the half maximal inhibitory concentration values ranging from 25.18?88.12 µM. Ludartin and lupeol were observed to have the most potent inhibitory effects. Based on the initially identified antiproliferative effects, these two compounds were evaluated for their effects on cell cycle phase distribution, DNA damage and their effects on mitochondrial membrane potential loss (??m). The two compounds induced DNA damage and mitochondrial membrane potential loss in MCF?7 cells. The results of the current study suggest that lupeol and ludartin, isolated from A. indica, produce anticancer effects by inducing DNA damage and a reduction of mitochondrial membrane potential, and may be used as potent anticancer agents, subsequent to further study. PMID:25434991

  11. Synthesis and regeneration of lead (IV) acetate

    SciTech Connect

    Boyle, T.J.; Al-Shareef, H.N.; Moore, G.J.

    1996-11-01

    Lead acetate [Pb(O{sub 2}CMe){sub 4}] was easily synthesized from a warm solution of Pb{sub 3}O{sub 4}, HO{sub 2}CMe and O(OCMe){sub 2} following literature preparations when the appropriate measures to minimize water contamination were followed. Furthermore, Pb(O{sub 2}CMe){sub 4} which has been decomposed (evidenced by the appearance of a purple color due to oxidation) can be regenerated using a similar preparatory route. Introduction of Pb(O{sub 2}CMe){sub 4} from the two routes outlined above into the IMO process for production of PZT thin films gave films with comparable ferroelectric properties to commercially available Pb(O{sub 2}CMe){sub 4} precursors. However, the freshly synthesized material yields PZT films with better properties compared to the recycled material.

  12. Heterogeneous catalyst for the production of acetic anhydride from methyl acetate

    DOEpatents

    Ramprasad, D.; Waller, F.J.

    1999-04-06

    This invention relates to a process for producing acetic anhydride by the reaction of methyl acetate, carbon monoxide, and hydrogen at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that contains an insoluble polymer having pendant quaternized phosphine groups, some of which phosphine groups are ionically bonded to anionic Group VIII metal complexes, the remainder of the phosphine groups being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled for consecutive runs without loss in activity. Bifunctional catalysts for use in carbonylating dimethyl ether are also provided.

  13. Heterogeneous catalyst for the production of acetic anhydride from methyl acetate

    DOEpatents

    Ramprasad, Dorai (Allentown, PA); Waller, Francis Joseph (Allentown, PA)

    1999-01-01

    This invention relates to a process for producing acetic anhydride by the reaction of methyl acetate, carbon monoxide, and hydrogen at elevated temperatures and pressures in the presence of an alkyl halide and a heterogeneous, bifunctional catalyst that contains an insoluble polymer having pendant quaternized phosphine groups, some of which phosphine groups are ionically bonded to anionic Group VIII metal complexes, the remainder of the phosphine groups being bonded to iodide. In contrast to prior art processes, no accelerator (promoter) is necessary to achieve the catalytic reaction and the products are easily separated from the catalyst by filtration. The catalyst can be recycled for consecutive runs without loss in activity. Bifunctional catalysts for use in carbonylating dimethyl ether are also provided.

  14. 21 CFR 522.2477 - Trenbolone acetate and estradiol.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL...trenbolone acetate and 24 mg estradiol (one implant consisting of 6 pellets, each pellet...trenbolone acetate and 4 mg estradiol) per implant dose. (B) 120 mg trenbolone...

  15. 21 CFR 522.2477 - Trenbolone acetate and estradiol.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... § 522.2477, see the List of CFR Sections Affected, which appears in the Finding Aids section of the... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Trenbolone acetate and estradiol. 522.2477 Section... § 522.2477 Trenbolone acetate and estradiol. (a) (b) Sponsors. See sponsors in § 510.600(c) of...

  16. Transition-Metal-Catalyzed Carbonylation of Methyl Acetate.

    ERIC Educational Resources Information Center

    Polichnowski, S. W.

    1986-01-01

    Presents a study of the rhodium-catalyzed, ioding-promoted carbonylation of methyl acetate. This study provides an interesting contrast between the carbonylation of methyl acetate and the carbonylation of methanol when similar rhodium/iodine catalyst systems are used. (JN)

  17. 21 CFR 520.1341 - Megestrol acetate tablets.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Megestrol acetate tablets. 520.1341 Section 520.1341 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.1341 Megestrol acetate tablets. (a) Specifications. Each...

  18. 21 CFR 522.2477 - Trenbolone acetate and estradiol.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... § 522.2477, see the List of CFR Sections Affected, which appears in the Finding Aids section of the... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Trenbolone acetate and estradiol. 522.2477 Section... § 522.2477 Trenbolone acetate and estradiol. (a) (b) Sponsors. See sponsors in § 510.600(c) of...

  19. Applications of space-electrophoresis in medicine. [for cellular separations in molecular biology

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1976-01-01

    The nature of electrophoresis is reviewed and potential advances realizable in the field of biology and medicine from a space electrophoresis facility are examined. The ground-based applications of electrophoresis: (1) characterization of an ionized species; (2) determination of the quantitative composition of a complex mixture; and (3) isolation of the components of a mixture, separation achieved on the basis of the difference in transport rates is reviewed. The electrophoresis of living cells is considered, touching upon the following areas: the separation of T and B lymphocytes; the genetic influence on mouse lymphocyte mobilities; the abnormal production of specific and monoclonal immunoproteins; and the study of cancer. Schematic diagrams are presented of three types of electrophoresis apparatus: the column assembly for the static electrophoresis experiment on the Apollo-Soyuz mission, the continuous flow apparatus used in the same mission and a miniaturized electrophoresis apparatus.

  20. Imaging Catalytic Surfaces by Multiplexed Capillary Electrophoresis With Absorption Detection

    SciTech Connect

    Michael Christodoulou

    2002-08-27

    A new technique for in situ imaging and screening heterogeneous catalysts by using multiplexed capillary electrophoresis with absorption detection was developed. By bundling the inlets of a large number of capillaries, an imaging probe can be created that can be used to sample products formed directly from a catalytic surface with high spatial resolution. In this work, they used surfaces made of platinum, iron or gold wires as model catalytic surfaces for imaging. Various shapes were recorded including squares and triangles. Model catalytic surfaces consisting of both iron and platinum wires in the shape of a cross were also imaged successfully. Each of the two wires produced a different electrochemical product that was separated by capillary electrophoresis. Based on the collected data they were able to distinguish the products from each wire in the reconstructed image.

  1. A simple gel electrophoresis method for separating polyhedral gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Kim, Suhee; Lee, Hye Jin

    2015-07-01

    In this paper, a simple approach to separate differently shaped and sized polyhedral gold nanoparticles (NPs) within colloidal solutions via gel electrophoresis is described. Gel running parameters for separating efficiently gold NPs including gel composition, added surfactant types and applied voltage were investigated. The plasmonic properties and physical structure of the separated NPs extracted from the gel matrix were then investigated using transmission electron microscopy (TEM) and UV-vis spectrophotometry respectively. Data analysis revealed that gel electrophoresis conditions of a 1.5 % agarose gel with 0.1 % sodium dodecyl sulfate (SDS) surfactant under an applied voltage of 100 V resulted in the selective isolation of ~ 50 nm polyhedral shaped gold nanoparticles. Further efforts are underway to apply the method to purify biomolecule-conjugated polyhedral Au NPs that can be readily used for NP-enhanced biosensing platforms.

  2. Effects of coating rectangular microscopic electrophoresis chamber with methylcellulose

    NASA Technical Reports Server (NTRS)

    Plank, L. D.

    1985-01-01

    One of the biggest problems in obtaining high accuracy in microscopic electrophoresis is the parabolic flow of liquid in the chamber due to electroosmotic backflow during application of the electric field. In chambers with glass walls the source of polarization leading to electroosmosis is the negative charge of the silicare and other ions that form the wall structure. It was found by Hjerten, who used a rotating 3.0 mm capillary tube for free zone electrophoresis, that precisely neutralizing this charge was extremely difficult, but if a neutral polymer matrix (formaldehyde fixed methylcellulose) was formed over the glass (quartz) wall the double layer was displaced and the viscosity at the shear plane increased so that electroosmotic flow could be eliminated. Experiments were designed to determine the reliability with which methylcellulose coating of the Zeiss Cytopherometer chamber reduced electroosmotic backflow and the effect of coating on the accuracy of cell electrophoretic mobility (EPN) determinations. Fixed rat erythrocytes (RBC) were used as test particles.

  3. Separation of DNA restriction fragments using capillary electrophoresis

    SciTech Connect

    Chan, K.C.; Whang, Chenwen; Yeung, E.S. )

    1993-01-01

    Gel-filled and non-gel' capillary electrophoresis (CE) have been applied to the separation of various DNA restriction fragments. 30% HydroLink gel, polymerized inside a 75[mu]m i.d. fused-silica capillary, was used in the gel-filled CE. Primary results show that the HL capillary gel was simple to cast, and its stability was reasonably good under the running conditions. In the non-gel CE experiment, a buffer containing the sieving additive hydroxypropylmethyl cellulose was used to affect the size-dependent separation. The use of GC capillaries eliminates the inconvenience of separately coating the capillary walls for efficient non-gel separation. Finally, the authors demonstrate that it is feasible to detect native DNA fragments using indirect fluorometry in non-gel capillary electrophoresis.

  4. On the importance of hydrodynamic interactions in polyelectrolyte electrophoresis

    E-print Network

    Kai Grass; Christian Holm

    2008-08-19

    The effect of hydrodynamic interactions on the free-solution electrophoresis of polyelectrolytes is investigated with coarse-grained molecular dynamics simulations. By comparing the results to simulations with switched-off hydrodynamic interactions, we demonstrate their importance in modelling the experimentally observed behaviour. In order to quantify the hydrodynamic interactions between the polyelectrolyte and the solution, we present a novel way to estimate its effective charge. We obtain an effective friction that is different from the hydrodynamic friction obtained from diffusion measurements. This effective friction is used to explain the constant electrophoretic mobility for longer chains. To further emphasize the importance of hydrodynamic interactions, we apply the model to end-labeled free-solution electrophoresis.

  5. A novel nanospray capillary zone electrophoresis/mass spectrometry interface.

    PubMed

    Hsieh, F; Baronas, E; Muir, C; Martin, S A

    1999-01-01

    The high resolution of capillary zone electrophoresis/mass spectrometry (CZE/MS) offers a promising technique to characterize biomolecules in pharmaceutical and biotechnology industries. A novel capillary zone electrophoresis/electrospray ionization time-of-flight mass spectrometry (CZE/ESI-TOFMS) interface was designed in this study to successfully integrate ESI-TOFMS, nanospray, and CZE for biomolecular identification. The interface offers a novel way to take advantage of the high resolution separation achieved during CZE and the detection sensitivity of nanospray ESI-MS. The results showed mixtures of peptides were highly resolved within a few minutes (each CZE electropherogram of a peptide is 2-3 seconds). The novel CZE/ESI-TOFMS interface may simultaneously provide sensitivity, data acquisition speed, mass range, and mass resolution while maintaining resolution of the CZE separation. PMID:9921690

  6. Omniphobic Membrane for Robust Membrane Distillation

    SciTech Connect

    Lin, SH; Nejati, S; Boo, C; Hu, YX; Osuji, CO; Ehmelech, M

    2014-11-01

    In this work, we fabricate an omniphobic microporous membrane for membrane distillation (MD) by modifying a hydrophilic glass fiber membrane with silica nanoparticles followed by surface fluorination and polymer coating. The modified glass fiber membrane exhibits an anti-wetting property not only against water but also against low surface tension organic solvents that easily wet a hydrophobic polytetrafluoroethylene (PTFE) membrane that is commonly used in MD applications. By comparing the performance of the PTFE and omniphobic membranes in direct contact MD experiments in the presence of a surfactant (sodium dodecyl sulfate, SDS), we show that SDS wets the hydrophobic PTFE membrane but not the omniphobic membrane. Our results suggest that omniphobic membranes are critical for MD applications with feed waters containing surface active species, such as oil and gas produced water, to prevent membrane pore wetting.

  7. Oxidation of Indole-3-acetic Acid and Oxindole-3-acetic Acid to 2,3-Dihydro-7-hydroxy-2-oxo-1H Indole-3-acetic Acid-7?-O-?-d-Glucopyranoside in Zea mays Seedlings 1

    PubMed Central

    Nonhebel, Heather M.; Bandurski, Robert S.

    1984-01-01

    Radiolabeled oxindole-3-acetic acid was metabolized by roots, shoots, and caryopses of dark grown Zea mays seedlings to 2,3-dihydro-7-hydroxy-2-oxo-1H indole-3-acetic acid-7?-O-?-d-glycopyranoside with the simpler name of 7-hydroxyoxindole-3-acetic acid-glucoside. This compound was also formed from labeled indole-3-acetic acid supplied to intact seedlings and root segments. The glucoside of 7-hydroxyoxindole-3-acetic acid was also isolated as an endogenous compound in the caryopses and shoots of 4-day-old seedlings. It accumulates to a level of 4.8 nanomoles per plant in the kernel, more than 10 times the amount of oxindole-3-acetic acid. In the shoot it is present at levels comparable to that of oxindole-3-acetic acid and indole-3-acetic acid (62 picomoles per shoot). We conclude that 7-hydroxyoxindole-3-acetic acid-glucoside is a natural metabolite of indole-3-acetic acid in Z. mays seedlings. From the data presented in this paper and in previous work, we propose the following route as the principal catabolic pathway for indole-3-acetic acid in Zea seedlings: Indole-3-acetic acid ? Oxindole-3-acetic acid ? 7-Hydroxyoxindole-3-acetic acid ? 7-Hydroxyoxindole-3-acetic acid-glucoside. PMID:11540902

  8. Oxidation of indole-3-acetic acid and oxindole-3-acetic acid to 2,3-dihydro-7-hydroxy-2-oxo-1H indole-3-acetic acid-7'-O-beta-D-glucopyranoside in Zea mays seedlings

    NASA Technical Reports Server (NTRS)

    Nonhebel, H. M.; Bandurski, R. S.

    1984-01-01

    Radiolabeled oxindole-3-acetic acid was metabolized by roots, shoots, and caryopses of dark grown Zea mays seedlings to 2,3-dihydro-7-hydroxy-2-oxo-1H indole-3-acetic acid-7'-O-beta-D-glycopyranoside with the simpler name of 7-hydroxyoxindole-3-acetic acid-glucoside. This compound was also formed from labeled indole-3-acetic acid supplied to intact seedlings and root segments. The glucoside of 7-hydroxyoxindole-3-acetic acid was also isolated as an endogenous compound in the caryopses and shoots of 4-day-old seedlings. It accumulates to a level of 4.8 nanomoles per plant in the kernel, more than 10 times the amount of oxindole-3-acetic acid. In the shoot it is present at levels comparable to that of oxindole-3-acetic acid and indole-3-acetic acid (62 picomoles per shoot). We conclude that 7-hydroxyoxindole-3-acetic acid-glucoside is a natural metabolite of indole-3-acetic acid in Z. mays seedlings. From the data presented in this paper and in previous work, we propose the following route as the principal catabolic pathway for indole-3-acetic acid in Zea seedlings: Indole-3-acetic acid --> Oxindole-3-acetic acid --> 7-Hydroxyoxindole-3-acetic acid --> 7-Hydroxyoxindole-3-acetic acid-glucoside.

  9. Sheathless interface for coupling capillary electrophoresis with mass spectrometry

    SciTech Connect

    Wang, Chenchen; Tang, Keqi; Smith, Richard D.

    2014-06-17

    A sheathless interface for coupling capillary electrophoresis (CE) with mass spectrometry is disclosed. The sheathless interface includes a separation capillary for performing CE separation and an emitter capillary for electrospray ionization. A portion of the emitter capillary is porous or, alternatively, is coated to form an electrically conductive surface. A section of the emitter capillary is disposed within the separation capillary, forming a joint. A metal tube, containing a conductive liquid, encloses the joint.

  10. Microchip Capillary Electrophoresis with Electrochemical Detection for Monitoring Environmental Pollutants

    SciTech Connect

    Chen, Gang; Lin, Yuehe; Wang, Joseph

    2006-01-15

    This invited paper reviews recent advances and the key strategies in microchip capillary electrophoresis (CE) with electrochemical detection (ECD) for separating and detecting a variety of environmental pollutants. The subjects covered include the fabrication of microfluidic chips, sample pretreatments, ECD, typical applications of microchip CE with ECD in environmental analysis, and future prospects. It is expected that microchip CE-ECD will become a powerful tool in the environmental field and will lead to the creation of truly portable devices.

  11. Electrohydrodynamics and other hydrodynamic phenomena in continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.

    1982-01-01

    The process known as continuous flow electrophoresis employs an electric field to separate the constituents of particulate samples suspended in a liquid. Complications arise because the electric field generates temperature gradients due to Joule heating and derives an electrohydrodynamic crossflow. Several aspects of the flow are discussed including entrance effects, hydrodynamic stability and a flow restructuring due to the combined effects of buoyancy and the crossflow.

  12. Lipid membranes for membrane proteins.

    PubMed

    Kukol, Andreas

    2015-01-01

    The molecular dynamics (MD) simulation of membrane proteins requires the setup of an accurate representation of lipid bilayers. This chapter describes the setup of a lipid bilayer system from scratch using generally available tools, starting with a definition of the lipid molecule POPE, generation of a lipid bilayer, energy minimization, MD simulation, and data analysis. The data analysis includes the calculation of area and volume per lipid, deuterium order parameters, self-diffusion constant, and the electron density profile. PMID:25330959

  13. Plasma membrane vesicles isolated from epimastigote forms of Trypanosoma cruzi.

    PubMed

    da Silveira, J F; Abrahamsohn, P A; Colli, W

    1979-01-19

    Membrane vesicles can be obtained from epimastigote forms of Trypansoma cruzi by incubating cells with either cross-linking reagents or acid pH. Acetate, phtalate or citrate, at pH 4.0, but not at higher pH values, were able to induce plasma membrane vesiculation. Vesicles have been purified by sucrose density centrifugation and their membrane origin was demonstrated by the following criteria: (a) Vesicles are 5--10 times richer in protein-bound iodine when they are prepared from cells previously labeled with 131I by the lactoperoxidase catalyzed reaction. (b) Electron microscopy of vesiculating cells shows physical continuity between cell plasma membrane and vesicle membrane. (c) Antibodies prepared against purified vesicles are able to agglutinate epimastigote forms of T. cruzi with sera dilutions up to 1 : 256 to 1 : 512. (d) Freeze-fracture studies of the purified vesicles have shown images of faces P and E compatible with known images of the intact cell plasma membrane. Typical preparations of acetate vesicles present the following characteristics: total carbohydrate : protein=1.5--2.0; orcinol : protein-0.07 and absence of diphenylamine reaction. Vesicles contain 0.2--0.5% and 0.3--1.0% of the total homogenate protein and carbohydrate, respectively. The presence of 10 major protein bands and 30--50-fold enrichment of the four sugar-containing macromolecules present in epimastigote forms of T. cruzi have been demonstrated in these preparations. PMID:365244

  14. Measurement of the rates of oxindole-3-acetic acid turnover, and indole-3-acetic acid oxidation in Zea mays seedlings

    NASA Technical Reports Server (NTRS)

    Nonhebel, H. M.; Bandurski, R. S. (Principal Investigator)

    1986-01-01

    Oxindole-3-acetic acid is the principal catabolite of indole-3-acetic acid in Zea mays seedlings. In this paper measurements of the turnover of oxindole-3-acetic acid are presented and used to calculate the rate of indole-3-acetic acid oxidation. [3H]Oxindole-3-acetic acid was applied to the endosperm of Zea mays seedlings and allowed to equilibrate for 24 h before the start of the experiment. The subsequent decrease in its specific activity was used to calculate the turnover rate. The average half-life of oxindole-3-acetic acid in the shoots was found to be 30 h while that in the kernels had an average half-life of 35h. Using previously published values of the pool sizes of oxindole-3-acetic acid in shoots and kernels from seedlings of the same age and variety, and grown under the same conditions, the rate of indole-3-acetic acid oxidation was calculated to be 1.1 pmol plant-1 h-1 in the shoots and 7.1 pmol plant-1 h-1 in the kernels.

  15. Integration of Microdialysis Sampling and Microchip Electrophoresis with Electrochemical Detection

    PubMed Central

    Mecker, Laura C.; Martin, R. Scott

    2009-01-01

    Here we describe the fabrication, optimization, and application of a microfluidic device that integrates microdialysis (MD) sampling, microchip electrophoresis (ME), and electrochemical detection (EC). The manner in which the chip is produced is reproducible and enables the fixed alignment of the MD/ME and ME/EC interfaces. Poly(dimethylsiloxane) (PDMS) -based valves were used for the discrete injection of sample from the hydrodynamic MD dialysate stream into a separation channel for analysis with ME. To enable the integration of ME with EC detection, a palladium decoupler was used to isolate the high voltages associated with electrophoresis from micron-sized carbon ink detection electrodes. Optimization of the ME/EC interface was needed to allow the use of biologically appropriate perfusate buffers containing high salt content. This optimization included changes in the fabrication procedure, increases in the decoupler surface area, and a programmed voltage shutoff. The ability of the MD/ME/EC system to sample a biological system was demonstrated by using a linear probe to monitor the stimulated release of dopamine from a confluent layer of PC 12 cells. To our knowledge, this is the first report of a microchip-based system that couples microdialysis sampling with microchip electrophoresis and electrochemical detection. PMID:19551945

  16. A poly-methylmethacrylate electrophoresis microchip with sample preconcentrator

    NASA Astrophysics Data System (ADS)

    Lin, Yu-Cheng; Ho, Hsiao-Ching; Tseng, Chien-Kai; Hou, Shao-Qin

    2001-05-01

    A microstructure on poly-methylmethacrylate (PMMA) for sample concentration and electrophoresis was fabricated. This microfabricated structure was able to increase the detection signal and lower the amount of sample used in electrophoretic analysis. The thin-film electrode located at the T-intersection of the sample injection and separation channels provides the current path for the injection channel, but restrains the DNA molecules from passing through. This can accumulate DNA molecules and increase the concentration before performing the electrophoretic analysis. This microstructure was fabricated using krypton fluoride (KrF) excimer laser photo-ablation and fusion bonding techniques. The excimer laser photo-ablation performs rapid prototyping with great flexibility in design changes. The PMMA material is much cheaper than other materials, for example glass and silicon, used in capillary electrophoresis and concentration. The applied electrical field was 300 V cm-1 for the DNA concentration in this microstructure. Experiments show that the DNA concentration was saturated within 200 s after the DNA molecules first reached the injection tee. The DNA fragments can be concentrated up to five times greater than samples without a concentrator at the injection tee. The separation results also demonstrated that the detected signal intensities of the separated DNA fragments in the tee-type chip with a sample preconcentrator were five times greater than that obtained in a conventional cross-type capillary electrophoresis chip with an identical initial sample concentration.

  17. Automated Lab-on-a-Chip Electrophoresis System

    NASA Technical Reports Server (NTRS)

    Willis, Peter A.; Mora, Maria; Greer, Harold F.; Fisher, Anita M.; Bryant, Sherrisse

    2012-01-01

    Capillary electrophoresis is an analytical technique that can be used to detect and quantify extremely small amounts of various biological molecules. In the search for biochemical traces of life on other planets, part of this search involves an examination of amino acids, which are the building blocks of life on Earth. The most sensitive method for detecting amino acids is the use of laser induced fluorescence. However, since amino acids do not, in general, fluoresce, they first must be reacted with a fluorescent dye label prior to analysis. After this process is completed, the liquid sample then must be transported into the electrophoresis system. If the system is to be reused multiple times, samples must be added and removed each time. In typical laboratories, this process is performed manually by skilled human operators using standard laboratory equipment. This level of human intervention is not possible if this technology is to be implemented on extraterrestrial targets. Microchip capillary electrophoresis (CE) combined with laser induced fluorescence detection (LIF) was selected as an extremely sensitive method to detect amino acids and other compounds that can be tagged with a fluorescent dye. It is highly desirable to package this technology into an integrated, autonomous, in situ instrument capable of performing CE-LIF on the surface of an extraterrestrial body. However, to be fully autonomous, the CE device must be able to perform a large number of sample preparation and analysis operations without the direct intervention of a human.

  18. Recent applications of microchip electrophoresis to biomedical analysis.

    PubMed

    Nuchtavorn, Nantana; Suntornsuk, Worapot; Lunte, Susan M; Suntornsuk, Leena

    2015-09-10

    Many separation methods have been developed for biomedical analysis, including chromatographic (e.g. high performance liquid chromatography (HPLC) and gas chromatography (GC)) and electrophoretic methods (e.g. gel electrophoresis and capillary electrophoresis (CE)). Among these techniques, CE provides advantages in terms of high separation efficiency, simplicity, low sample and solvent volume consumption, short analysis time and applicability to a wide range of biomedically important substances. Microchip electrophoresis (ME) is a miniaturized platform of CE and is now considered as a simpler and more convenient alternative, which has demonstrated potential in analytical chemistry. High-throughput, cost-effective and portable analysis systems can be developed using ME. The current review describes different separation modes and detectors that have been employed in ME to analyze various classes of biomedical analytes (e.g. pharmaceuticals and related substances, nucleic acids, amino acids, peptides, proteins, antibodies and antigens, carbohydrates, cells, cell components and lysates). Recent applications (during 2010-2014) in these areas are presented in tables and some significant findings are highlighted. PMID:25840947

  19. Visualization of DNA molecules in time during electrophoresis

    NASA Technical Reports Server (NTRS)

    Lubega, Seth

    1991-01-01

    For several years individual DNA molecules have been observed and photographed during agarose gel electrophoresis. The DNA molecule is clearly the largest molecule known. Nevertheless, the largest molecule is still too small to be seen using a microscope. A technique developed by Morikawa and Yanagida has made it possible to visualize individual DNA molecules. When these long molecules are labeled with appropriate fluorescence dyes and observed under a fluorescence microscope, although it is not possible to directly visualize the local ultrastructure of the molecules, yet because they are long light emitting chains, their microscopic dynamical behavior can be observed. This visualization works in the same principle that enables one to observe a star through a telescope because it emits light against a dark background. The dynamics of individual DNA molecules migrating through agarose matrix during electrophoresis have been described by Smith et al. (1989), Schwartz and Koval (1989), and Bustamante et al. (1990). DNA molecules during agarose gel electrophoresis advance lengthwise thorough the gel in an extended configuration. They display an extension-contraction motion and tend to bunch up in their leading ends as the 'heads' find new pores through the gel. From time to time they get hooked on obstacles in the gel to form U-shaped configurations before they resume their linear configuration.

  20. Procedures for two-dimensional electrophoresis of proteins

    SciTech Connect

    Tollaksen, S.L.; Giometti, C.S.

    1996-10-01

    High-resolution two-dimensional gel electrophoresis (2DE) of proteins, using isoelectric focusing in the first dimension and sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) in the second, was first described in 1975. In the 20 years since those publications, numerous modifications of the original method have evolved. The ISO-DALT system of 2DE is a high-throughput approach that has stood the test of time. The problem of casting many isoelectric focusing gels and SDS-PAGE slab gels (up to 20) in a reproducible manner has been solved by the use of the techniques and equipment described in this manual. The ISO-DALT system of two-dimensional gel electrophoresis originated in the late 1970s and has been modified many times to improve its high-resolution, high-throughput capabilities. This report provides the detailed procedures used with the current ISO-DALT system to prepare, run, stain, and photograph two-dimensional gels for protein analysis.

  1. Hybridization thermodynamics of DNA oligonucleotides during microchip capillary electrophoresis.

    PubMed

    Wynne, Thomas M; McCallum, Christopher; Del Bonis-O'Donnell, Jackson Travis; Crisalli, Pete; Pennathur, Sumita

    2015-03-01

    Capillary electrophoresis (CE) is a powerful analytical tool for performing separations and characterizing properties of charged species. For reacting species during a CE separation, local concentrations change leading to nonequilibrium conditions. Interpreting experimental data with such nonequilibrium reactive species is nontrivial due to the large number of variables involved in the system. In this work we develop a COMSOL multiphysics-based numerical model to simulate the electrokinetic mass transport of short interacting ssDNAs in microchip capillary electrophoresis. We probe the importance of the dissociation constant, K(D), and the concentration of DNA on the resulting observed mobility of the dsDNA peak, ?(w), by using a full sweep of parametric simulations. We find that the observed mobility is strongly dependent on the DNA concentration and K(D), as well as ssDNA concentration, and develop a relation with which to understand this dependence. Furthermore, we present experimental microchip capillary electrophoresis measurements of interacting 10 base ssDNA and its complement with changes in buffer ionic strength, DNA concentration, and DNA sequence to vary the system equilibria. We then compare our results to thermodynamically calculated K(D) values. PMID:25634338

  2. Human lymphocyte polymorphisms detected by quantitative two-dimensional electrophoresis.

    PubMed Central

    Goldman, D; Merril, C R

    1983-01-01

    A survey of 186 soluble lymphocyte proteins for genetic polymorphism was carried out utilizing two-dimensional electrophoresis of 14C-labeled phytohemagglutinin (PHA)-stimulated human lymphocyte proteins. Nineteen of these proteins exhibited positional variation consistent with independent genetic polymorphism in a primary sample of 28 individuals. Each of these polymorphisms was characterized by quantitative gene-dosage dependence insofar as the heterozygous phenotype expressed approximately 50% of each allelic gene product as was seen in homozygotes. Patterns observed were also identical in monozygotic twins, replicate samples, and replicate gels. The three expected phenotypes (two homozygotes and a heterozygote) were observed in each of 10 of these polymorphisms while the remaining nine had one of the homozygous classes absent. The presence of the three phenotypes, the demonstration of gene-dosage dependence, and our own and previous pedigree analysis of certain of these polymorphisms supports the genetic basis of these variants. Based on this data, the frequency of polymorphic loci for man is: P = 19/186 = .102, and the average heterozygosity is .024. This estimate is approximately 1/3 to 1/2 the rate of polymorphism previously estimated for man in other studies using one-dimensional electrophoresis of isozyme loci. The newly described polymorphisms and others which should be detectable in larger protein surveys with two-dimensional electrophoresis hold promise as genetic markers of the human genome for use in gene mapping and pedigree analyses. Images Fig. 1 Fig. 3 PMID:6577787

  3. [Affinity capillary electrophoresis for screening proteins interacting with domoic acid].

    PubMed

    Wang, Xiaoqian; Gao, Tie; Hong, Zhuan; Qu, Feng

    2015-07-01

    Domoic acid (DA) is a biological neurotoxin that causes amnesic shellfish poisoning. Study of the interactions between DA and important functional proteins contributes to understand the toxicity mechanism of DA to biological macromolecules. In this paper, the interactions between DA and nine important proteins in plasma, intestine and mitochondria were qualitatively compared by affinity capillary electrophoresis. Proteins were used as affinity ligands while DA as the affinity receptor. Proteins with the concentrations of 0, 0.2, 0.4, 0.6, 0.8 ?mol/L were added in the electrophoresis buffer and the migration times of 0.2 mg/mL DA were detected. Then the linear graphs of the variation of DA mobility ratio (?M) with the protein mass concentration (L) were drawn. According to the slope value, the relative strength of the interactions between DA and proteins was compared. The results showed that six proteins can interact with DA and the relative strength order was human thrombin > cytochrome C trypsin immunoglobulin E (Ig E) ? ribonuclease A > ? exonuclease, while ferritin, transferrin and lectin had no affinity with DA. With the advantages of high efficiency, fast analysis and less sample consumption, affinity capillary electrophoresis is a convenient method for screening DA target proteins, which will provide basic information for the toxic mechanism and defence of DA. PMID:26672193

  4. Microfabricated capillary electrophoresis amino acid chirality analyzer for extraterrestrial exploration

    NASA Technical Reports Server (NTRS)

    Hutt, L. D.; Glavin, D. P.; Bada, J. L.; Mathies, R. A.

    1999-01-01

    Chiral separations of fluorescein isothiocyanate-labeled amino acids have been performed on a microfabricated capillary electrophoresis chip to explore the feasibility of using such devices to analyze for extinct or extant life signs in extraterrestrial environments. The test system consists of a folded electrophoresis channel (19.0 cm long x 150 microns wide x 20 microns deep) that was photolithographically fabricated in a 10-cm-diameter glass wafer sandwich, coupled to a laser-excited confocal fluorescence detection apparatus providing subattomole sensitivity. Using a sodium dodecyl sulfate/gamma-cyclodextrin pH 10.0 carbonate electrophoresis buffer and a separation voltage of 550 V/cm at 10 degrees C, baseline resolution was observed for Val, Ala, Glu, and Asp enantiomers and Gly in only 4 min. Enantiomeric ratios were determined for amino acids extracted from the Murchison meteorite, and these values closely matched values determined by HPLC. These results demonstrate the feasibility of using microfabricated lab-on-a-chip systems to analyze extraterrestrial samples for amino acids.

  5. Microchip electrophoresis: a method for high-speed SNP detection

    PubMed Central

    Schmalzing, Dieter; Belenky, Alexei; Novotny, Mark A.; Koutny, Lance; Salas-Solano, Oscar; El-Difrawy, Sameh; Adourian, Aram; Matsudaira, Paul; Ehrlich, Daniel

    2000-01-01

    As a trial practical application, we have applied optimized microfabricated electrophoresis devices, combined with enzymatic mutation detection methods, to the determination of single nucleotide polymorphism (SNP) sites in the p53 suppressor gene. Using clinical samples, we have achieved robust assays with quality factors as good as conventional electrophoresis in ~100 s. This is 10 and 50 times faster than capillary and slab gel electrophoresis, respectively. The method was highly accurate with an average error of mutation site measurement of only ±5 bp. No clean-up of the digestion mixtures was needed prior to injection. This greatly simplifies sample handling relative to capillary instruments, which is important for high-throughput screening applications. Following identification, absolute mutation determination of the screened samples was achieved in a second microdevice optimized for four-color DNA sequencing. Total run time was 25 min in this second device and sequencing data were in full agreement with ABI Prism® 377 sequencing runs which required 3.5 h. The tandem application of microdevices for location then full characterization of SNPs appears to confirm many of the improvements claimed for future application of microdevices in practical scaled screening for mutational analysis. PMID:10756210

  6. Capillary zone electrophoresis-mass spectrometry of peptides and proteins

    SciTech Connect

    Loo, J.A.; Udseth, H.R.; Smith, R.D.

    1989-05-01

    Capillary zone electrophoresis (CZE) is attracting extensive attention as a fast, high resolution analytical and micro-preparative separations technique for systems of biological interest. In zone electrophoresis, a column is filled with a single electrolyte having a specific conductivity. The mixture of substances to be separated is applied as a narrow band to the head of a buffer filled column in a band whose width is much less than the length of the column and at a concentration too low to affect the buffer conductivity. An electric field is then applied across the length of the column and the individual substances migrate and separate according to their net electrophoretic velocities. Zone electrophoresis carried out in small diameter (<100 ..mu..m) fused silica capillaries is a relatively new approach to the high resolution separation of aqueous samples. Very small volume samples (picoliter range) with separation efficiencies on the order of 10/sup 6/ theoretical plates for amino acids have been achieved. The method can be further enhanced by the dynamic combination of detection sensitivity and selectivity offered by mass spectrometry (MS). The on-line marriage of mass spectrometry to CZE is accomplished by an atmospheric pressure electrospray ionization source interface. Our research efforts have demonstrated that proteins with MW's greater than 100 kDa can be analyzed using a conventional quadrupole mass spectrometer with an upper m/z limit of only 1700. 6 refs.

  7. Analysis of hemoglobin electrophoresis results and physicians investigative practices in Saudi Arabia

    PubMed Central

    Mehdi, Syed Riaz; Al Dahmash, Badr Abdullah

    2013-01-01

    BACKGROUND AND OBJECTIVES: Riyadh and central province falls in a moderate prevalent zone of hemoglobinopathies in Saudi Arabia. However, it has been observed that the physicians working in Saudi Arabia invariably advise all cases of anemia for hemoglobin electrophoresis (HE). The present work was carried out to study the yield of the HE in Riyadh and the investigative practices of the physicians advising HE. SETTINGS AND DESIGN: The study was carried out in the hospitals of King Saud University from 2009 to 2011 in order to assess the yield of HE in referred cases of clinical anemia. MATERIALS AND METHODS: A total of 1073 cases divided in two groups of males and females had undergone complete blood count and red blood cell morphology. Cellulose acetate HE was performed and all the positive results were reconfirmed on the high performance liquid chromatography (HPLC). The results were analyzed for the type of hemoglobinopathies. For statistical analysis Statistical Package for Social Sciences 15 version (SPSS Inc., Chicago, IL, USA) was used. RESULTS: A total of 405 males and 668 females blood samples were included in the present study. 116 (28.5%) males and 167 (25%) females showed an abnormal pattern on HE. The incidence of beta thalassemia trait was higher in females while sickle cell trait was predominantly seen in males. Red cell indices were reduced considerably in thalassemias, but were unaffected in sickle cell disorders, except those which had concurrent alpha trait. The total yield of HE was 26.6% which was much less than expected. CONCLUSION: The physicians are advised to rule out iron deficiency and other common causes of anemia before investigating the cases for hemoglobinopathies, which employs time consuming and expensive tests of HE and HPLC. PMID:24339548

  8. Inhibitors of thermally induced burn incidents – characterization by microbiological procedure, electrophoresis, SEM, DSC and IR spectroscopy.

    PubMed

    Pielesz, Anna; Machnicka, Alicja; Gaw?owski, Andrzej; Fabia, Janusz; Sarna, Ewa; Binia?, W?odzimierz

    2015-07-01

    Differential scanning calorimetry (DSC) and thermogravimetric (TGA) investigations, acetate electrophoresis (CAE), Fourier-transform infrared spectrometry (FTIR), scanning electron microscopy (SEM) analysis and microbiological procedures were all carried out after heating the samples to a temperature sufficient for simulating a burn incident. In particular, the purpose of the present study was to analyze the effect of antioxidants, such as fucoidan from brown seaweed and flame-retardant cyclic organophosphates and phosphonates, on an organic chicken skin that gets changed by a burn incident. DSC was considered to be a useful tool in assessing in vitro temperature-mediated cross-linking; an innovative analytical conclusion was obtained from the experimentation described in the paper. FTIR tests revealed that heating a dry organic chicken skin to the boiling point leads to the disappearance of a wide band in the 1650-1550 cm(-1) area or the conversion of a band, which may be attributed to the intermolecular ?-sheet aggregates. Fucoidan from brown seaweed and flame-retardant cyclic organophosphates and phosphonates probably bind with the collagen that is changed by the burn (in addition to the influence of antioxidant solutions on samples of a blank or not boiled organic chicken skin) incident forming a polymer film with the collagen of the chicken skin surface (SEM analysis), decreasing the aggregation process and native collagen recovery. Good bacteriostatic properties were determined for fucoidan samples from brown seaweed and flame-retardant cyclic organophosphates and phosphonates against the pathogenic bacteria Escherichia coli and Staphylococcus aureus. Thus, it was observed that the fucoidan incorporated into collagen films can be used as a therapeutically active biomaterial that speeds up the wound-healing process. PMID:26029873

  9. Fast determination of harmala alkaloids in edible algae by capillary electrophoresis mass spectrometry.

    PubMed

    Tascon, Marcos; Benavente, Fernando; Sanz-Nebot, Victoria M; Gagliardi, Leonardo G

    2015-05-01

    The use of algae as a foodstuff is rapidly expanding worldwide from the East Asian countries, where they are also used for medical care. Harmala alkaloids (HAlk) are a family of bioactive compounds found in the extracts of some plants, including wakame (Undaria pinnatifida), an edible marine invasive algae. HAlks are based on a characteristic ?-carboline structure with at least one amino ionizable group. In this work, we report the successful separation of a mixture of six HAlks (harmine, harmaline, harmol, harmalol, harmane, and norharmane) by capillary electrophoresis ion-trap mass spectrometry (CE-IT-MS) in less than 8 min. Optimum separation in fused-silica capillaries and detection sensitivity in positive-ion mode were achieved using a background electrolyte (BGE) with 25 mmol L(-1) ammonium acetate (pH 7.8) and 10% (v/v) methanol, and a sheath liquid with 60:40 (v/v) isopropanol-water and 0.05% (v/v) formic acid. The separation method was validated in terms of linearity, limits of detection and quantification, repeatability, and reproducibility. Later, a sample pretreatment was carefully optimized to determine HAlks in commercial wakame samples with excellent recovery and repeatability. For the complex wakame extracts, the MS-MS fragmentation patterns of the different HAlks were useful to ensure a reliable identification. The complete procedure was validated using the standard-addition calibration method, determining matrix effects on the studied compounds. Harmalol, harmine, and harmaline were naturally present in the samples and were quantified at very low concentrations, ranging from 7 to 24 ?g kg(-1) dry algae. PMID:25749794

  10. Application of capillary electrophoresis combined with a modified BCR sequential extraction for estimating of distribution of selected trace metals in PM2.5 fractions of urban airborne particulate matter.

    PubMed

    Dabek-Zlotorzynska, Ewa; Kelly, Meghan; Chen, Heidi; Chakrabarti, Chuni L

    2005-03-01

    The capillary electrophoresis (CE) method combined with a sequential extraction was applied to determine the distribution of Fe, Zn, Cu, Mn and Cd in urban ambient air PM2.5 samples. PM2.5 was collected on Teflon filters with dichotomous sampler, and the modified extraction procedure following the BCR leaching procedure was used to chemically fractionate metals into "easily exchangeable" with water, "acid extractable" with 0.11 mol/l acetic acid, "reducible" with 0.1 mol/l hydroxylamine hydrochloride acidified to pH 2.0 with nitric acid, and "oxidisable" with oxidation by 8.8 mol/l hydrogen peroxide (H2O2) followed by extraction with 1.0 mol/l ammonium acetate. Based on the obtained results it was concluded that the application of the studied methodology provides chemical fractionation data that reflect the general sources and potential health hazards of the studied metals. PMID:15686754

  11. Identification of acetic acid bacteria in traditionally produced vinegar and mother of vinegar by using different molecular techniques.

    PubMed

    Yetiman, Ahmet E; Kesmen, Zülal

    2015-07-01

    Culture-dependent and culture-independent methods were combined for the investigation of acetic acid bacteria (AAB) populations in traditionally produced vinegars and mother of vinegar samples obtained from apple and grape. The culture-independent denaturing gradient gel electrophoresis (DGGE) analysis, which targeted the V7-V8 regions of the 16S rRNA gene, showed that Komagataeibacter hansenii and Komagataeibacter europaeus/Komagataeibacter xylinus were the most dominant species in almost all of the samples analyzed directly. The culture-independent GTG5-rep PCR fingerprinting was used in the preliminary characterization of AAB isolates and species-level identification was carried out by sequencing of the 16S rRNA gene, 16S-23S rDNA internally transcribed to the spacer (ITS) region and tuf gene. Acetobacter okinawensis was frequently isolated from samples obtained from apple while K. europaeus was identified as the dominant species, followed by Acetobacter indonesiensis in the samples originating from grape. In addition to common molecular techniques, real-time PCR intercalating dye assays, including DNA melting temperature (Tm) and high resolution melting analysis (HRM), were applied to acetic acid bacterial isolates for the first time. The target sequence of ITS region generated species-specific HRM profiles and Tm values allowed discrimination at species level. PMID:25828705

  12. From paper electrophoresis to computer-supported interpretation of capillary electrophoresis--clinical plasma protein analysis in Malmö, Sweden.

    PubMed

    Carlson, J

    2001-11-01

    Protein analyses have been used in Malmö as a routine clinical diagnostic tool since 1953. Most serum samples are submitted for "protein profiles" including capillary zone electrophoresis and rate immune nephelometric quantification of nine proteins (five in urines), although analysis of single proteins may be requested. Standardization between laboratories in our region has been greatly improved by automation, CRM 470 calibration and external quality assurance. We are further extending standardization by developing computer supported interpretations using a program with improved user interface and graphical representation of electrophoretic curves superimposed upon a shaded reference interval. Programming is underway to provide complete automatic interpretation of these curves. Together, capillary electrophoresis (with access to mathematical analysis) and immunochemical quantifications allow a highly automated process accessible to further digital analysis and automated interpretation. Rapid, cost-effective and standardized analysis of serum protein profiles should improve the diagnostic evaluation of many categories of patients. PMID:11831616

  13. 76 FR 32366 - Determination That ORLAAM (Levomethadyl Acetate Hydrochloride) Oral Solution, 10 Milligrams...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-06

    ...DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and...Levomethadyl Acetate Hydrochloride) Oral Solution, 10 Milligrams/Milliliter...acetate hydrochloride (HCl)) oral solution, 10 milligrams (mg...for levomethadyl acetate HCl oral solution, 10 mg/mL, if...

  14. 40 CFR 180.1258 - Acetic acid; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...2011-07-01 2011-07-01 false Acetic acid; exemption from the requirement...From Tolerances § 180.1258 Acetic acid; exemption from the requirement...residues of the biochemical pesticide acetic acid when used as a...

  15. Lipopolymer gradient diffusion in supported bilayer membranes.

    PubMed

    Zhang, Huai-Ying; Hill, Reghan J

    2011-03-01

    We measure the gradient diffusion coefficient of a model lipopolymer in supported lipid bilayer membranes from Fourier-transform post-electrophoresis relaxation. The experiments and accompanying quantitative interpretation furnish the concentration dependence of the gradient diffusion coefficient. In striking contrast to the recent measurements of the self-diffusion coefficient from fluorescence recovery after photobleaching, the lipopolymer gradient diffusion coefficient increases with concentration. We interpret the enhancement at small but finite concentrations using the Scalettar-Abney-Owicki (SAO) statistical mechanical theory (1988) and the Bussell-Koch-Hammer (BKH) hydrodynamic theory (1995), which are customarily adopted to model membrane protein dynamics. The SAO theory furnishes an effective disc radius and soft repulsive interaction radius that are comparable to the Flory radius of the unperturbed polyethylene glycol chains. On the other hand, the BKH theory predicts a gradient diffusion coefficient that decreases with disc/membrane protein concentration. Thus, in contrast to membrane proteins, we conclude that lipopolymer hydrodynamic interactions are weak because the principal disturbances are in the low-viscosity aqueous phase. Accordingly, lipopolymer interactions are dominated by thermodynamic interactions among polymer chains. Interestingly, our experiments suggest that increasing (decreasing) the polymer molecular weight should increase (decrease) the relaxation rate of lipopolymer concentration fluctuations. PMID:20702448

  16. Rotating Field Gel Electrophoresis (RAGE) is a variation on Pulsed Field Gel Electrophoresis (PFGE) and gives similar results. We use equipment that was only briefly marketed by

    E-print Network

    Cross, George

    1 RAGE Rotating Field Gel Electrophoresis (RAGE) is a variation on Pulsed Field Gel Electrophoresis lower cost than BioRad PFGE equipment. Plugs for RAGE/PFGE Starting material for BF: 100 ml (50 ml. Set up gel tray (drawer underneath RAGE apparatus): 4 pieces. Make sure that the rotating circular

  17. The study on pervaporation behaviors of dilute organic solution through PDMS/PTFE composite membrane.

    PubMed

    Zhang, Wei-dong; Sun, Wei; Yang, Jing; Ren, Zhong-qi

    2010-01-01

    As an energy-efficient alternative to distillation, pervaporation has been widely combined with fermentation to remove organic compounds from their dilute solutions in a fermentation broth. In this work, the organic permselective composite membrane is prepared by coating polydimethylsiloxane (PDMS) cross-linked with n-heptane on the substrate of polytetrafluoroethylene(PTFE) membrane. The separation behavior is studied in different dilute organic solutions, which include acetone dilute solution, butanone dilute solution, cyclohexanone dilute solution, ethanol dilute solution, isopropanol dilute solution, n-butyl alcohol dilute solution, acetic acid dilute solution, and ethyl acetate dilute solution. Most of these solutions are main reaction products or by-products from fermentation process. The effects of solubility of organics in the membrane, molecular weight, and polarity of the organics on the pervaporation performance are investigated. The effects of operating temperature and organic concentration in the feed solutions on the performance of composite membrane are studied as well. The experimental results show that molecular volume has less influence than solubility and molecular polarity for these organic solvent. The selectivity of PDMS membrane to ethyl acetate is relative high due to good solubility and diffusion of ethyl acetate molecules in polymer. PMID:19259826

  18. Formate and acetate in monsoon rainwater of Agra, India

    SciTech Connect

    Kumar, N.; Kulshrestha, U.C.; Saxena, A.; Kumari, K.M.; Srivastava, S.S. )

    1993-03-20

    Formate and acetate concentrations were estimated using ion chromatography in 19 precipitation samples collected on an eventwise basis during the monsoon season (July through September), 1991, at Dayalbagh, Agra. Volume-weighted average (VWA) concentrations for formate and acetate were 5.8 and 6.55 [mu]molL[sup [minus]1], respectively. The VWA hydrogen ion concentration was 0.084 [mu]eq L[sub [minus]1] (pH 7.07) and the correlation coefficient between the two ions was 0.85. The average formate to acetate ratio was low (0.88), possibly due to an increase in acetate contribution from direct emissions associated with heavy vehicular traffic load and/or indirect acetate formation by alkaline hydrolysis of PAN. Widespread local use of biomass as a domestic fuel may also contribute acetate. In 4 of the 19 precipitation events studied, higher values of both species were recorded. Contributions from soil in addition to vegetation, were suspected in these samples. Inputs from soil and combustion activities were supported by correlations among formate, acetate and Ca[sup 2+] (terrigenous species), K[sup +], SO[sub 4][sup 2[minus

  19. A Search for Methyl Acetate in Hot Cores

    NASA Astrophysics Data System (ADS)

    Kelley, Matthew; Braakman, Rogier; Blake, Geoffrey

    2006-10-01

    We propose to search for methyl acetate, CH3COOCH3, in high mass hot cores. Methyl acetate is possibly synthesized through multiple reaction pathways from molecules previously detected in hot cores, most notably from acetic acid and methanol via esterification. Esterification, beyond the formation of methyl formate, has not yet been observed in the ISM. The project is already underway in nothern sources based on millimeter data from the Caltech Submillimeter Observatory (CSO). We hope to study the chemically rich southern source G327.3 (decl B1950=-54.49'15.6"), amongst others, using MOPRA and to search for acetic acid and methyl acetate. Observations are supported by laboratory studies of methyl acetate in the 3 mm and 1 mm and successful spectral fitting by the Blake group at Caltech. If detected, methyl acetate, consisting of 11 atoms, would be one of the larger complex organic molecules detected in the interstellar medium and could point to previously unconsidered reaction mechanisms.

  20. The Effects of Acetate Buffer Concentration on Lysozyme Solubility

    NASA Technical Reports Server (NTRS)

    Forsythe, Elizabeth L.; Pusey, Marc L.

    1996-01-01

    The micro-solubility column technique was employed to systematically investigate the effects of buffer concentration on tetragonal lysozyme solubility. While keeping the NaCl concentrations constant at 2%, 3%, 4%, 5% and 7%, and the pH at 4.0, we have studied the solubility of tetragonal lysozyme over an acetate buffer concentration range of 0.01M to 0.5M as a function of temperature. The lysozyme solubility decreased with increasing acetate concentration from 0.01M to 0.1M. This decrease may simply be due to the net increase in solvent ionic strength. Increasing the acetate concentration beyond 0.1M resulted in an increase in the lysozyme solubility, which reached a peak at - 0.3M acetate concentration. This increase was believed to be due to the increased binding of acetate to the anionic binding sites of lysozyme, preventing their occupation by chloride. In keeping with the previously observed reversal of the Hoffmeister series for effectiveness of anions in crystallizing lysozyme, acetate would be a less effective precipitant than chloride. Further increasing the acetate concentration beyond 0.3M resulted in a subsequent gradual decrease in the lysozyme solubility at all NaCl concentrations.

  1. Crystal structure of a mixed solvated form of amoxapine acetate

    PubMed Central

    Bhardwaj, Rajni M.; Raval, Vishal; Oswald, Iain D. H.; Florence, Alastair J.

    2015-01-01

    The mixed solvated salt 4-(2-chloro­dibenzo[b,f][1,4]oxazepin-11-yl)piperazin-1-ium acetate–acetic acid–cyclo­hexane (2/2/1), C17H17ClN3O+·C2H3O2 ?·C2H4O2·0.5C6H12, crystallizes with one mol­ecule of protonated amoxapine (AXPN), an acetate anion and a mol­ecule of acetic acid together with half a mol­ecule of cyclo­hexane. In the centrosymmetric crystal, both enanti­omers of the protonated AXPN mol­ecule stack alternatively along [001]. Acetate anions connect the AXPN cations through N—H?O hydrogen bonding in the [010] direction, creating a sheet lying parallel to (100). The acetic acid mol­ecules are linked to the acetate anions via O—H?O hydrogen bonds within the sheets. Within the sheets there are also a number of C—H?O hydrogen bonds present. The cyclo­hexane solvent mol­ecules occupy the space between the sheets. PMID:25878802

  2. Increased brain uptake and oxidation of acetate in heavy drinkers

    PubMed Central

    Jiang, Lihong; Gulanski, Barbara Irene; De Feyter, Henk M.; Weinzimer, Stuart A.; Pittman, Brian; Guidone, Elizabeth; Koretski, Julia; Harman, Susan; Petrakis, Ismene L.; Krystal, John H.; Mason, Graeme F.

    2013-01-01

    When a person consumes ethanol, the body quickly begins to convert it to acetic acid, which circulates in the blood and can serve as a source of energy for the brain and other organs. This study used 13C magnetic resonance spectroscopy to test whether chronic heavy drinking is associated with greater brain uptake and oxidation of acetic acid, providing a potential metabolic reward or adenosinergic effect as a consequence of drinking. Seven heavy drinkers, who regularly consumed at least 8 drinks per week and at least 4 drinks per day at least once per week, and 7 light drinkers, who consumed fewer than 2 drinks per week were recruited. The subjects were administered [2-13C]acetate for 2 hours and scanned throughout that time with magnetic resonance spectroscopy of the brain to observe natural 13C abundance of N-acetylaspartate (NAA) and the appearance of 13C-labeled glutamate, glutamine, and acetate. Heavy drinkers had approximately 2-fold more brain acetate relative to blood and twice as much labeled glutamate and glutamine. The results show that acetate transport and oxidation are faster in heavy drinkers compared with that in light drinkers. Our finding suggests that a new therapeutic approach to supply acetate during alcohol detoxification may be beneficial. PMID:23478412

  3. Computerized image analysis for acetic acid induced intraepithelial lesions

    NASA Astrophysics Data System (ADS)

    Li, Wenjing; Ferris, Daron G.; Lieberman, Rich W.

    2008-03-01

    Cervical Intraepithelial Neoplasia (CIN) exhibits certain morphologic features that can be identified during a visual inspection exam. Immature and dysphasic cervical squamous epithelium turns white after application of acetic acid during the exam. The whitening process occurs visually over several minutes and subjectively discriminates between dysphasic and normal tissue. Digital imaging technologies allow us to assist the physician analyzing the acetic acid induced lesions (acetowhite region) in a fully automatic way. This paper reports a study designed to measure multiple parameters of the acetowhitening process from two images captured with a digital colposcope. One image is captured before the acetic acid application, and the other is captured after the acetic acid application. The spatial change of the acetowhitening is extracted using color and texture information in the post acetic acid image; the temporal change is extracted from the intensity and color changes between the post acetic acid and pre acetic acid images with an automatic alignment. The imaging and data analysis system has been evaluated with a total of 99 human subjects and demonstrate its potential to screening underserved women where access to skilled colposcopists is limited.

  4. Methane Production and Syntrophic Acetate Oxidation in the Florida Everglades

    NASA Astrophysics Data System (ADS)

    Holmes, M. E.; Chanton, J.; Bae, H.; Ogram, A.

    2012-12-01

    Methane production pathways in the Florida Everglades are influenced by factors such as nutrient levels, H2 concentrations, and temperature. Syntrophic acetate oxidizers can outcompete methanogens for acetate when conditions are right (high temperatures and low H2). During syntrophic acetate oxidation (SAO), which becomes more exergonic with increasing temperature, acetate is oxidized to carbon dioxide and H2, which can be utilized to produce methane via CO2 reduction. Everglades soil from along a nutrient gradient was incubated at 25°C and 45°C. The shift to the CO2 reduction pathway for methane formation that would be expected in high temperature incubations due to SAO should result in a decrease in ?13C-CH4 and increase in ?2H-CH4. Instead, we observed higher ?13C and lower ?2H in the methane produced in high temperature incubations. The higher than expected ?13C may be partly explained by lower kinetic isotope effects caused by temperature. Coupling between the syntrophic acetate oxidizers and the CO2 reducers, whereby isotopically light hydrogen from acetate is used in methane formation could lower ?2H-CH4. Separate experiments using 13C-labelled acetate revealed that potential SAO activity is low in soils collected from the Everglades.

  5. Continuous flow electrophoresis: The effect of sample concentration on throughput and resolution in an upward flowing system

    NASA Technical Reports Server (NTRS)

    Jandebeur, T. S.

    1980-01-01

    The effect of sample concentration on throughput and resolution in a modified continuous particle electrophoresis (CPE) system with flow in an upward direction is investigated. Maximum resolution is achieved at concentrations ranging from 2 x 10 to the 8th power cells/ml to 8 x 10 to the 8th power cells/ml. The widest peak separation is at 2 x 10 to the 8th power cells/ml; however, the sharpest peaks and least overlap between cell populations is at 8 x 10 to the 8th power cells/ml. Apparently as a result of improved electrophoresis cell performance due to coasting the chamber with bovine serum albumin, changing the electrode membranes and rinse, and lowering buffer temperatures, sedimentation effects attending to higher concentrations are diminished. Throughput as measured by recovery of fixed cells is diminished at the concentrations judged most likely to yield satisfactory resolution. The tradeoff appears to be improved recovery/throughput at the expense of resolution.

  6. Microtubule-membrane interactions in cilia. I. Isolation and characterization of ciliary membranes from Tetrahymena pyriformis

    PubMed Central

    1980-01-01

    Tetrahymena ciliary membranes were prepared by four different techniques, and their protein composition was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), electron microscopy, and two-dimensional thin-layer peptide mapping. Extraction of the isolated cilia by nonionic detergent solubilized the ciliary membranes but left the axonemal microtubules and dyneine arms intact, as determined by quantitative electron microscopy. The proteins solubilized by detergent included a major 55,000-dalton protein, 1-3 high molecular weight proteins that comigrated, on SDS-PAGE, with the axonemal dynein, as well as several other proteins of 45,000-50,000 daltons. Each of the major proteins contained a small amount of carbohydrate, as determined by PAS-staining; no PAS-positive material was detected in the detergent-extracted axonemes. The major 55,000- dalton protein has proteins quite similar to those of tubulin, based on SDS-PAGE using three different buffer systems as well as two- dimensional maps of tryptic peptides from the isolated 55,000-dalton protein. To determine whether this tubulin-like protein was associated with the membrane or whether it was an axonemal or matrix protein released by detergent treatment, three different methods to isolate ciliary membrane vesicles were developed. The protein composition of each of these differetn vesicle preparations was the same as that of the detergent-solubilized material. These results suggest that a major ciliary membrane protein has properties similar to those of tubulin. PMID:6445909

  7. Magnetic Membrane System

    DOEpatents

    McElfresh, Michael W.; (Livermore, CA); Lucas, Matthew S.; (Pasadena, CA)

    2004-12-30

    The present invention provides a membrane with magnetic particles. In one embodiment the membrane is created by mixing particles in a non-magnetic base. The membrane may act as an actuator, a sensor, a pump, a valve, or other device. A magnet is operatively connected to the membrane. The magnet acts on and changes the shape of the membrane.

  8. Selecting a Roof Membrane.

    ERIC Educational Resources Information Center

    Waldron, Larry W.

    1990-01-01

    Offers a brief synopsis of the unique characteristics of the following roof membranes: (1) built-up roofing; (2) elastoplastic membranes; (3) modified bitumen membranes; (4) liquid applied membranes; and (5) metal roofing. A chart compares the characteristics of the raw membranes only. (MLF)

  9. Stable carbon isotope discrimination in rice field soil during acetate turnover by syntrophic acetate oxidation or acetoclastic methanogenesis

    NASA Astrophysics Data System (ADS)

    Conrad, Ralf; Klose, Melanie

    2011-03-01

    Rice fields are an important source for the greenhouse gas methane. In Italian rice field soil CH 4 is produced either by hydrogenotrophic and acetoclastic methanogenesis, or by hydrogenotrophic methanogenesis and syntrophic acetate oxidation when temperatures are below and above about 40-45 °C, respectively. In order to see whether these acetate consumption pathways differently discriminate the stable carbon isotopes of acetate, we measured the ? 13C of total acetate and acetate-methyl as well as the ? 13C of CO 2 and CH 4 in rice field soil that had been pre-incubated at 45 °C and then shifted to different temperatures between 25 and 50 °C. Acetate transiently accumulated to about 6 mM, which is about one-third of the amount of CH 4 produced, irrespective of the incubation temperature and the CH 4 production pathway involved. However, the patterns of ? 13C of the CH 4 and CO 2 produced were different at low (25, 30, 35 °C) versus high (40, 45, 50 °C) temperatures. These patterns were consistent with CH 4 being exclusively formed by hydrogenotrophic methanogenesis at high temperatures, and by a combination of acetoclastic and hydrogenotrophic methanogenesis at low temperatures. The patterns of ? 13C of total acetate and acetate-methyl were also different at high versus low temperatures, indicating the involvement of different pathways of production and consumption of acetate at the two temperature regimes. Isotope fractionation during consumption of the methyl group of acetate was more pronounced at low ( ? = 1.010-1.025) than at high ( ? = 1.0-1.01) temperatures indicating that acetoclastic methanogenesis exhibits a stronger isotope effect than syntrophic acetate oxidation. Small amounts of propionate also transiently accumulated and were analyzed for ? 13C. The ? 13C values slightly increased (by about 10‰) during production and consumption of propionate, but were not affected by incubation temperature. Collectively, our results showed distinct isotope discrimination for different paths of acetate (and propionate) production and consumption, albeit differences were only small, and discrimination between methanogenic and syntrophic acetate consumption in nature may be difficult to detect.

  10. Selective Cross-Coupling of Organic Halides with Allylic Acetates

    PubMed Central

    Anka-Lufford, Lukiana L.; Prinsell, Michael R.

    2012-01-01

    A general protocol for the coupling of haloarenes with a variety of allylic acetates is presented. Strengths of the method are a tolerance for electrophilic (ketone, aldehyde) and acidic (sulfonamide, trifluoroacetamide) substrates and the ability to couple with a variety of substituted allylic acetates. Secondary alkyl bromides can also be allylated under slightly modified conditions, demonstrating the generality of the approach. Finally, the coupling of a reactive vinyl halide could be achieved by the use of a very hindered ligand and more reactive, branched allylic acetates. PMID:23095043

  11. Clostridiumm ljungdahlii, an anaerobic ethanol and acetate producing microorganism

    DOEpatents

    Gaddy, J.L.; Clausen, E.C.

    1992-12-22

    A newly discovered microorganism was isolated in a biologically pure culture and designated Clostridium ljungdahlii, having the identifying characteristics of ATCC No. 49587. Cultured in an aqueous nutrient medium under anaerobic conditions, this microorganism is capable of producing ethanol and acetate from CO and H[sub 2]O and/or CO[sub 2] and H[sub 2] in synthesis gas. Under optimal growth conditions, the microorganism produces acetate in preference to ethanol. Conversely, under non-growth conditions, ethanol production is favored over acetate. 3 figs.

  12. Clostridiumm ljungdahlii, an anaerobic ethanol and acetate producing microorganism

    DOEpatents

    Gaddy, James L. (Fayetteville, AR); Clausen, Edgar C. (Fayetteville, AR)

    1992-01-01

    A newly discovered microorganism was isolated in a biologically pure culture and designated Clostridium ljungdahlii, having the identifying characteristics of ATCC No. 49587. Cultured in an aqueous nutrient medium under anaerobic conditions, this microorganism is capable of producing ethanol and acetate from CO and H.sub.2 O and/or CO.sub.2 and H.sub.2 in synthesis gas. Under optimal growth conditions, the microorganism produces acetate in preference to ethanol. Conversely, under non-growth conditions, ethanol production is favored over acetate.

  13. Experimental and theoretical investigation of the stability of stepwise pH gradients in continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Kuhn, Reinhard; Wagner, Horst; Mosher, Richard A.; Thormann, Wolfgang

    1987-01-01

    Isoelectric focusing in the continuous flow mode can be more quickly and economically performed by admitting a stepwise pH gradient composed of simple buffers instead of uniform mixtures of synthetic carrier ampholytes. The time-consuming formation of the pH gradient by the electric field is thereby omitted. The stability of a three-step system with arginine - morpholinoethanesulfonic acid/glycylglycine - aspartic acid is analyzed theoretically by one-dimensional computer simulation as well as experimentally at various flow rates in a continuous flow apparatus. Excellent agreement between experimental and theoretical data was obtained. This metastable configuration was found to be suitable for focusing of proteins under continuous flow conditions. The influence of various combinations of electrolytes and membranes between electrophoresis chamber and electrode compartments is also discussed.

  14. Phosphorylated lymphocyte plasma-membrane proteins.

    PubMed Central

    Johnstone, A P; DuBois, J H; Crumpton, M J

    1981-01-01

    Lymphocytes were labelled by incubation with [32P]Pi and their plasma membranes isolated. Analysis by one-dimensional and two-dimensional gel electrophoresis revealed a small number of strongly phosphorylated polypeptides. Two of these were especially prominent; they had molecular weights of about 52000 and 90000, were acidic and were apparently not glycosylated. Similar patterns were obtained for quiescent T- and B-lymphocytes from different species and for cultured lymphoblastoid cells, although the relative amounts of the labelled polypeptides varied. Immunoprecipitation analyses of the detergent-solubilized 32P-labelled plasma membranes indicated that the glycosylated polypeptide of the human major transplantation (HLA-A and HLA-B) antigens and its mouse and pig counterparts are phosphorylated. In contrast, no phosphorylation of the membrane-associated immunoglobulin, the mouse Thy-1 antigen or the human HLA-DRw(Ia) antigen was detected. The phosphorylation patterns of human peripheral blood and nude-mouse spleen lymphocytes did not change during the period 5-30min after mitogen stimulation. Therefore a change in the phosphorylation of plasma-membrane protein(s) is probably not an early biochemical event in the initiation of T-lymphocyte and B-lymphocyte growth, although a rapid transient change cannot be ruled out. Similar plasma-membrane phosphorylation patterns were also obtained by incubating the purified plasma membrane with [gamma-32P]ATP. The phosphorylation of the 90000-mol.wt. polypeptide was particularly rapid and was stimulated by the addition of cyclic AMP. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:7305985

  15. Capillary zone electrophoresis for U(VI) and short chain carboxylic acid sorption studies on silica and rutile.

    PubMed

    Sladkov, Vladimir; Zhao, Yujia; Mercier-Bion, Florence

    2011-02-15

    Capillary zone electrophoresis was used to study the uranyl and short chain carboxylic acid sorption on silica and rutile. The separation and the simultaneous determination (in a single run) of a number of short chain carboxylic acids (oxalic, formic, acetic and propionic) and U(VI) with direct UV detection is developed for the analysis of solutions after the sorption experiments. The reverse polarity mode is used (the injection is performed at the negative end). The matrix effect of Si(IV) (possible silica dissolution product) and perchlorate (added for constant ionic strength in sorption experiments) on the separation of U(VI) and organic acids is investigated. The influence of methanol addition in carrier electrolyte on the separation selectivity of given analytes is also studied. Under the chosen conditions (carbonate buffer (ionic strength of 0.1M), pH 9.8, 0.15 mM of tetradecyltrimethylammonium bromide, 25% (v/v) of methanol) the calibration curves are plotted. They are linear in two ranges of concentration from ?1×10(-5) to ?1×10(-3) M for oxalate, acetate, propionate, U(VI) and ?1×10(-4) to ?1×10(-3) for formate. The accuracy of the procedure is checked by the "added-found" method in simulation solutions. The relative standard deviations of the concentrations found are within the range of 1-10% and the recovery is in the range of 90-115%. This method is applied for the analysis of aqueous samples issued from sorption experiments on silica and rutile. The obtained results indicate that the given organic acids decrease uranium sorption both on silica and rutile. These experiments demonstrate that short chain carboxylic acids can influence the mobility and the chemistry of U(VI) in the environment. PMID:21238757

  16. Membrane Systems in Cyanobacteria

    SciTech Connect

    Liberton, Michelle L.; Pakrasi, Himadri B.

    2008-01-01

    Cyanobacteria are photosynthetic prokaryotes with highly differentiated membrane systems. In addition to a Gram-negative-type cell envelope with plasma membrane and outer membrane separated by a periplasmic space, cyanobacteria have an internal system of thylakoid membranes where the fully functional electron transfer chains of photosynthesis and respiration reside. The presence of different membrane systems lends these cells a unique complexity among bacteria. Cyanobacteria must be able to reorganize the membranes, synthesize new membrane lipids, and properly target proteins to the correct membrane system. The outer membrane, plasma membrane, and thylakoid membranes each have specialized roles in the cyanobacterial cell. Understanding the organization, functionality, protein composition and dynamics of the membrane systems remains a great challenge in cyanobacterial cell biology.

  17. Water dispersible microbicidal cellulose acetate phthalate film

    PubMed Central

    Neurath, A Robert; Strick, Nathan; Li, Yun-Yao

    2003-01-01

    Background Cellulose acetate phthalate (CAP) has been used for several decades in the pharmaceutical industry for enteric film coating of oral tablets and capsules. Micronized CAP, available commercially as "Aquateric" and containing additional ingredients required for micronization, used for tablet coating from water dispersions, was shown to adsorb and inactivate the human immunodeficiency virus (HIV-1), herpesviruses (HSV) and other sexually transmitted disease (STD) pathogens. Earlier studies indicate that a gel formulation of micronized CAP has a potential as a topical microbicide for prevention of STDs including the acquired immunodeficiency syndrome (AIDS). The objective of endeavors described here was to develop a water dispersible CAP film amenable to inexpensive industrial mass production. Methods CAP and hydroxypropyl cellulose (HPC) were dissolved in different organic solvent mixtures, poured into dishes, and the solvents evaporated. Graded quantities of a resulting selected film were mixed for 5 min at 37°C with HIV-1, HSV and other STD pathogens, respectively. Residual infectivity of the treated viruses and bacteria was determined. Results The prerequisites for producing CAP films which are soft, flexible and dispersible in water, resulting in smooth gels, are combining CAP with HPC (other cellulose derivatives are unsuitable), and casting from organic solvent mixtures containing ?50 to ?65% ethanol (EtOH). The films are ?100 µ thick and have a textured surface with alternating protrusions and depressions revealed by scanning electron microscopy. The films, before complete conversion into a gel, rapidly inactivated HIV-1 and HSV and reduced the infectivity of non-viral STD pathogens >1,000-fold. Conclusions Soft pliable CAP-HPC composite films can be generated by casting from organic solvent mixtures containing EtOH. The films rapidly reduce the infectivity of several STD pathogens, including HIV-1. They are converted into gels and thus do not have to be removed following application and use. In addition to their potential as topical microbicides, the films have promise for mucosal delivery of pharmaceuticals other than CAP. PMID:14617380

  18. A study of cell electrophoresis as a means of purifying growth hormone secreting cells

    NASA Technical Reports Server (NTRS)

    Plank, Lindsay D.; Hymer, W. C.; Kunze, M. Elaine; Marks, Gary M.; Lanham, J. Wayne

    1983-01-01

    Growth hormone secreting cells of the rat anterior pituitary are heavily laden with granules of growth hormone and can be partialy purified on the basis of their resulting high density. Two methods of preparative cell electrophoresis were investigated as methods of enhancing the purification of growth hormone producing cells: density gradient electrophoresis and continuous flow electrophoresis. Both methods provided a two- to four-fold enrichment in growth hormone production per cell relative to that achieved by previous methods. Measurements of electrophoretic mobilities by two analytical methods, microscopic electrophoresis and laser-tracking electrophoresis, revealed very little distinction between unpurified anterior pituitary cell suspensions and somatotroph-enriched cell suspensions. Predictions calculated on the basis of analytical electrophoretic data are consistent with the hypothesis that sedimentation plays a significant role in both types of preparative electrophoresis and the electrophoretic mobility of the growth hormone secreting subpopulation of cells remains unknown.

  19. Monitoring refolding of tailspike endorhamnosidase using capillary electrophoresis-laser induced tryptophan fluorescence

    SciTech Connect

    Jensen, P.K.; Lee, Cheng S.; King, J.A.

    1997-12-31

    The use of capillary electrophoresis equipped with laser-induced tryptophan fluorescence detection is presented for monitoring the refolding pathway of phage P22 tailspike endorhamnosidase. Upon initiation of refolding, tailspike polypeptides rapidly fold into structured monomeric intermediates with a high content of secondary structure. These monomeric species associate to form the triple-chain defined folding intermediates, the protrimers. Conversion of the protrimer into the native, sodium dodecyl sulfate (SDS) resistant tailspike protein is the rate-limiting step in the refolding pathway. Refolding kinetics and yield measured by capillary electrophoresis are in good agreement with those obtained via native gel electrophoresis, SDS polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence spectrophotometry. To enhance separation resolution between protrimer and native protein in capillary electrophoresis, the use of poly(ethylene oxide) is investigated for the introduction of a sieving separation mechanism. The increased viscosity of the electrophoresis buffer may also play a role in resolution enhancement.

  20. 21 CFR 522.960b - Flumethasone acetate solution.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS § 522.960b Flumethasone acetate solution. (a) Specifications....

  1. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...per cubic centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol...preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1 milligram of citric acid; water for injection...

  2. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...per cubic centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol...preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1 milligram of citric acid; water for injection...

  3. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...per cubic centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol...preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1 milligram of citric acid; water for injection...

  4. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...per cubic centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol...preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1 milligram of citric acid; water for injection...

  5. SOLVENT EXTRACTION OF WASTEWATERS FROM ACETIC-ACID MANUFACTURE

    EPA Science Inventory

    Solvent extraction was evaluated as a potential treatment method for wastewaters generated during the manufacture of acetic acid. Possible goals for an extraction process were considered. For the wastewater samples studied, extraction appeared to be too expensive to be practical ...

  6. Fragrance material review on 2-hydroxy-2-phenylethyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 2-hydroxy-2-phenylethyl acetate when used as a fragrance ingredient is presented. 2-Hydroxy-2-phenylethyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 2-hydroxy-2-phenylethyl acetate was evaluated then summarized and includes physical properties data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414656

  7. Fragrance material review on 2-(p-tolyloxy)ethyl acetate.

    PubMed

    McGinty, D; Letizia, C S; Api, A M

    2012-09-01

    A toxicologic and dermatologic review of 2-(p-tolyloxy)ethyl acetate when used as a fragrance ingredient is presented. 2-(p-tolyloxy)ethyl acetate is a member of the fragrance structural group aryl alkyl alcohol simple acid esters (AAASAE). The AAASAE fragrance ingredients are prepared by reacting an aryl alkyl alcohol with a simple carboxylic acid (a chain of 1-4 carbons) to generate formate, acetate, propionate, butyrate, isobutyrate and carbonate esters. This review contains a detailed summary of all available toxicology and dermatology papers that are related to this individual fragrance ingredient and is not intended as a stand-alone document. Available data for 2-(p-tolyloxy)ethyl acetate were evaluated, then summarized, and includes physical properties data. A safety assessment of the entire AAASAE will be published simultaneously with this document. Please refer to Belsito et al. (2012) for an overall assessment of the safe use of this material and all AAASAE in fragrances. PMID:22414652

  8. Degradation by acetic acid for crystalline Si photovoltaic modules

    NASA Astrophysics Data System (ADS)

    Masuda, Atsushi; Uchiyama, Naomi; Hara, Yukiko

    2015-04-01

    The degradation of crystalline Si photovoltaic modules during damp-heat test was studied using some test modules with and without polymer film insertion by observing electrical and electroluminescence properties and by chemical analyses. Acetic acid generated by the hydrolysis decomposition of ethylene vinyl acetate used as an encapsulant is the main origin of degradation. The change in electroluminescence images is explained on the basis of the corrosion of electrodes by acetic acid. On the other hand, little change was observed at the pn junction even after damp-heat test for a long time. Therefore, carrier generation occurs even after degradation; however, such generated carriers cannot be collected owing to corrosion of electrodes. The guiding principle that module structure and module materials without saving acetic acid into the modules was obtained.

  9. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol (as preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1...

  10. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol (as preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1...

  11. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol (as preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1...

  12. 21 CFR 522.960b - Flumethasone acetate injection.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... centimeter: 2 milligrams of flumethasone acetate; 20 milligrams of propylene glycol; 9 milligrams of benzyl alcohol (as preservative); 8 milligrams of sodium chloride; 1 milligram of polysorbate 80; 0.1...

  13. Highly Enantioselective Hydrogenation of Enol Acetates Catalyzed by

    E-print Network

    Zhang, Xumu

    enol acetates. Catalytic asymmetric synthesis presents one of most powerful methods for accessing Although atropisomeric biaryl diphosphines such as BI- NAP,1,4 BIPHEMP,1,5 and MeO-BIPHEP1,5 have been used

  14. Electronic interactions between gold films and mn12-acetate 

    E-print Network

    Means, Joel Lewis

    2009-05-15

    Interactions between Mn12–acetate molecular magnets and thin gold films have been explored in light of the theory of weak localization. Low-temperature measurements of the magnetoresistance of gold films of varying thicknesses, with and without...

  15. 21 CFR 584.200 - Ethyl alcohol containing ethyl acetate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...gallons of 100 percent ethyl acetate. It is used in accordance with good feeding practices in ruminant feed supplements as a source of added energy. [46 FR 52333, Oct. 27, 1981, as amended at 72 FR 41620, July 31,...

  16. Cosolvent gel-like materials from partially hydrolyzed poly(vinyl acetate)s and borax.

    PubMed

    Angelova, Lora V; Terech, Pierre; Natali, Irene; Dei, Luigi; Carretti, Emiliano; Weiss, Richard G

    2011-09-20

    A gel-like, high-viscosity polymeric dispersion (HVPD) based on cross-linked borate, partially hydrolyzed poly(vinyl acetate) (xPVAc, where x is the percent hydrolysis) is described. Unlike hydro-HVPDs prepared from poly(vinyl alcohol) (PVA) and borate, the liquid portion of these materials can be composed of up to 75% of an organic cosolvent because of the influence of residual acetate groups on the polymer backbone. The effects of the degree of hydrolysis, molecular weight, polymer and cross-linker concentrations, and type and amount of organic cosolvent on the rheological and structural properties of the materials are investigated. The stability of the systems is explored through rheological and melting-range studies. (11)B NMR and small-angle neutron scattering (SANS) are used to probe the structure of the dispersions. The addition of an organic liquid to the xPVAc-borate HVPDs results in a drastic increase in the number of cross-linked borate species as well as the agglomeration of the polymer into bundles. These effects result in an increase in the relaxation time and thermal stability of the networks. The ability to make xPVAc-borate HVPDs with very large amounts of and rather different organic liquids, with very different rheological properties that can be controlled easily, opens new possibilities for applications of PVAc-based dispersions. PMID:21848256

  17. Recent progress in preparation and application of microfluidic chip electrophoresis

    NASA Astrophysics Data System (ADS)

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Yuan, Hua; Peng, Qiaohong; Tian, Chao

    2015-05-01

    Since its discovery in 1990, microfluidic chip electrophoresis (MCE) has allowed the development of applications with small size, fast analysis, low cost, high integration density and automatic level, which are easy to carry and have made commercialization efficient. MCE has been widely used in the areas of environmental protection, biochemistry, medicine and health, clinical testing, judicial expertise, food sanitation, pharmaceutical checking, drug testing, agrochemistry, biomedical engineering and life science. As one of the foremost fields in the research of capillary electrophoresis, MCE is the ultimate frontier to develop the miniaturized, integrated, automated all-in-one instruments needed in modern analytical chemistry. By adopting the advanced technologies of micro-machining, lasers and microelectronics, and the latest research achievements in analytical chemistry and biochemistry, the sampling, separation and detection systems of commonly used capillary electrophoresis are integrated with high densities onto glass, quartz, silicon or polymer wafers to form the MCE, which can finish the analysis of multi-step operations such as injection, enrichment, reaction, derivatization, separation, and collection of samples in a portable, efficient and super high speed manner. With reference to the different technological achievements in this area, the latest developments in MCE are reviewed in this article. The preparation mechanisms, surface modifications, and properties of different materials in MCE are compared, and the different sampling, separation and detection systems in MCE are summarized. The performance of MCE in analysis of fluorescent substance, metallic ion, sugar, medicine, nucleic acid, DNA, amino acid, polypeptide and protein is discussed, and the future direction of development is forecast.

  18. Coalification process waste water reusability: separation of organics by membranes

    SciTech Connect

    Bhattacharyya, D.; Kermode, R.I.; Dickinson, R.L.

    1983-02-01

    The overall objective of this investigation is to provide a critical evaluation of the current information concerning coal-gasification wastewaters and to establish experimentally the extent of separation of phenolics and polynuclear aromatic hydrocarbons (from single and multi-solute synthetic systems) by low-and high-pressure composite membranes. The compounds selected for experimental investigation were: phenol, O-cresol, 2,3-dimethylphenol, catechol, resorcinol, 2-naphthol, naphthalene, and indole. The development of membrane separation processes is gaining considerable importance because of the feasibility of simultaneous removal of organics and inorganic dissolved solids. Cellulose-acetate membranes developed for desalination processes show no rejection of phenolics; however, recently developed thin-film, noncellulosic composite membranes (even at low-pressure operation) may be useful in gasification wastewater reuse schemes. 24 references, 11 figures, 5 tables.

  19. Western Blotting Using PVDF Membranes and Its Downstream Applications.

    PubMed

    Komatsu, Setsuko

    2015-01-01

    Western blotting using polyvinylidene difluoride (PVDF) membranes is one of the most popular techniques for detection and characterization of proteins. If this technique is combined with immunodetection, the behavior of a particular protein can be clarified. On the other hand, if it is combined with Edman sequencing, the primary structure of the protein can be determined. A protein sample is transferred from an SDS-polyacrylamide gel electrophoresis (PAGE) gel onto a PVDF membrane by electroblotting. The membrane carrying the protein is either used for immunodetection or protein sequencing. SDS-PAGE followed by Western blotting combined with immunodetection using antibodies can easily detect protein behavior in crude protein mixtures. Furthermore, two-dimensional PAGE followed by Western blotting and Edman sequencing allows effective sequence determination of crude protein mixtures that may not be easily purified by conventional column chromatography. PMID:26044005

  20. Methods for Mapping of Interaction Networks Involving Membrane Proteins

    SciTech Connect

    Hooker, Brian S.; Bigelow, Diana J.; Lin, Chiann Tso

    2007-11-23

    Numerous approaches have been taken to study protein interactions, such as tagged protein complex isolation followed by mass spectrometry, yeast two-hybrid methods, fluorescence resonance energy transfer, surface plasmon resonance, site-directed mutagenesis, and crystallography. Membrane protein interactions pose significant challenges due to the need to solubilize membranes without disrupting protein-protein interactions. Traditionally, analysis of isolated protein complexes by high-resolution 2D gel electrophoresis has been the main method used to obtain an overall picture of proteome constituents and interactions. However, this method is time consuming, labor intensive, detects only abundant proteins and is not suitable for the coverage required to elucidate large interaction networks. In this review, we discuss the application of various methods to elucidate interactions involving membrane proteins. These techniques include methods for the direct isolation of single complexes or interactors as well as methods for characterization of entire subcellular and cellular interactomes.

  1. Highly conductive ethylene-vinyl acetate copolymer/carbon nanotube paper for lightweight and flexible supercapacitors

    NASA Astrophysics Data System (ADS)

    Zhang, Zishou; Wang, Wang; Li, Cheng; Wei, Lin; Chen, Xiaojun; Tong, Yexiang; Mai, Kancheng; Lu, Xihong

    2014-02-01

    Herein, we developed a highly conductive and lightweight membrane substrate based on the ethylene-vinyl acetate copolymer (EVA), carbon nanotubes (CNTs) and lens wiping paper, and demonstrated its implementation as high-performance mechanical support for MnO2 in supercapacitors. The 45 s-MnO2/EVA/40 wt%CNT electrode exhibited a high capacitance (0.126 F cm-2 at 0.5 mA cm-2) with excellent flexibility. Furthermore, the fabricated devices based on these 45 s-MnO2/EVA/40 wt%CNT electrodes exhibited excellent flexibility and stability (85% of its initial capacitance retained after 800 bending cycles) and achieved a high energy density of 9.4 Wh kg-1 at 6780 W kg-1.

  2. The effect of uranyl acetate on human lymphoblastoid cells (RPMI 6410) and HeLa cells.

    PubMed Central

    Ghadially, F. N.; Yang-Steppuhn, S. E.; Lalonde, J. M.

    1982-01-01

    RPMI 6410 cells and HeLa cells were exposed to uranyl acetate. In RPMI 6410 cell cultures this produced single-membrane-bound presumably lysosomal bodies (called "uraniosomes") containing electron-dense crystals in the cultured cells and crystalline deposits in extracellular locations. Neither uraniosomes nor extracellular uranium deposits were found in HeLa cell cultures. All uraniosomes and extracellular uranium deposits analysed by electron-probed X-ray analysis were found to contain uranium, potassium and phosphorus. Traces of sulphur were detected in some but not all uraniosomes and extracellular uranium deposits. Traces of calcium were found in all extracellular uranium deposits and in some uraniosomes also. Images Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:7093141

  3. An intercomparison of measurement systems for vapor and particulate phase concentrations of formic and acetic acids

    NASA Astrophysics Data System (ADS)

    Keene, William C.; Talbot, Robert W.; Andreae, Meinrat O.; Beecher, Kristene; Berresheim, Harold

    1989-05-01

    During June 1986, eight systems for measuring vapor phase and four for measuring particulate phase concentrations of formic acid (HCOOH) and acetic acid (CH3COOH) were intercompared in central Virginia. HCOOH and CH3COOH vapors were sampled by condensate, mist, Chromosorb 103 GC resin, NaOH-coated annular denuders, NaOH-impregnated quartz filters, K2CO3 and NaCO3-impregnated cellulose filters, and Nylasorb membranes. Atmospheric aerosol was collected on Teflon and Nuclepore filters using both hi-vol and lo-vol systems to measure particulate phase concentrations. Performances of the mist chamber and K2CO3-impregnated filter techniques were evaluated using zero air and ambient air spiked with HCOOH(g) and CH3COOH(g), and formaldehyde from permeation sources. The advantages and drawbacks of these methods are reported and discussed.

  4. An intercomparison of measurement systems for vapor and particulate phase concentrations of formic and acetic acids

    NASA Technical Reports Server (NTRS)

    Keene, William C.; Talbot, Robert W.; Andreae, Meinrat O.; Beecher, Kristene; Berresheim, Harold

    1989-01-01

    During June 1986, eight systems for measuring vapor phase and four for measuring particulate phase concentrations of formic acid (HCOOH) and acetic acid (CH3COOH) were intercompared in central Virginia. HCOOH and CH3COOH vapors were sampled by condensate, mist, Chromosorb 103 GC resin, NaOH-coated annular denuders, NaOH-impregnated quartz filters, K2CO3 and NaCO3-impregnated cellulose filters, and Nylasorb membranes. Atmospheric aerosol was collected on Teflon and Nuclepore filters using both hi-vol and lo-vol systems to measure particulate phase concentrations. Performances of the mist chamber and K2CO3-impregnated filter techniques were evaluated using zero air and ambient air spiked with HCOOH(g) and CH3COOH(g), and formaldehyde from permeation sources. The advantages and drawbacks of these methods are reported and discussed.

  5. Contrast manifestation of alkaline phosphatase and 5'-nucleotidase in plasma membranes isolated from rat liver and ascites hepatoma.

    PubMed

    Ikehara, Y; Takahashi, K; Mansho, K; Eto, S; Kato, K

    1977-10-17

    1. Plasma membranes were isolated from ascites hepatoma AH-130 and rat livers with or without partial hepatectomy or bile duct ligation. Reciprocal manifestations of two marker enzymes for plasma membranes were observed in these membrane preparations; alkaline phosphatase activity was found much higher in the hepatoma membrane than in any preparations of the liver membranes, whereas 5'-nucleotidase activity was much lower in the former than in the latter. 2. Effects of lectins and anti-plasma membrane antiserum on these two marker enzymes were examined. The results showed that about 50% of apparent activity of 5'-nucleotidase found in the hepatoma membrane was exhibited by alkaline phosphatase. 3. Localizations of alkaline phosphatase and 5'-nucleotidase in polyacrylamide gels after electrophoresis were demonstrated using 5'-AMP and 5-Br, 4-Cl-indoxylphosphate as substrate. There was a difference in electrophoretic mobility between the alkaline phosphatase of the hepatoma and that of the liver. PMID:20952

  6. Microorganisms having enhanced resistance to acetate and methods of use

    DOEpatents

    Brown, Steven D; Yang, Shihui

    2014-10-21

    The present invention provides isolated or genetically modified strains of microorganisms that display enhanced resistance to acetate as a result of increased expression of a sodium proton antiporter. The present invention also provides methods for producing such microbial strains, as well as related promoter sequences and expression vectors. Further, the present invention provides methods of producing alcohol from biomass materials by using microorganisms with enhanced resistance to acetate.

  7. Fingerprint analysis of Flos Carthami by capillary electrophoresis.

    PubMed

    Sun, Yi; Guo, Tao; Sui, Yin; Li, Famei

    2003-07-25

    Capillary electrophoresis (CE) was employed in fingerprint analysis of Flos Carthami. A standardized procedure was used to develop the CE fingerprint. An electrophoretic profile of genuine Flos Carthami from Fengqiu, He'nan, China, was first established as the characteristic fingerprint. This profile was then used to identify and assess the consistency of the herb. A study with a limited number of samples from nine sources showed a fair consistency in their CE fingerprints with that of the genuine sample. Flos Carthami was well distinguished from Stigma Croci, a possible substitute in traditional Chinese medicine, and Flos Hemerocallis, a commercial adulterant, by comparing the fingerprints of each herb. PMID:12860022

  8. Separation of long RNA by agarose-formaldehyde gel electrophoresis.

    PubMed

    Mansour, Farrah H; Pestov, Dimitri G

    2013-10-01

    We describe a method to facilitate electrophoretic separation of high-molecular-weight RNA species, such as ribosomal RNAs and their precursors, on agarose-formaldehyde gels. Two alternative "pK-matched" buffer systems were substituted for the traditionally used Mops-based conductive medium. The key advantages include shortened run times, a 5-fold reduction in formaldehyde concentration, a significantly improved resolution of long RNAs, and consistency in separation. The new procedure has a streamlined work flow that helps to minimize errors and is broadly applicable to agarose gel electrophoresis of RNA samples and their subsequent analysis by Northern blotting. PMID:23800830

  9. Human muscle proteins: analysis by two-dimensional electrophoresis

    SciTech Connect

    Giometti, C.S.; Danon, M.J.; Anderson, N.G.

    1983-09-01

    Proteins from single frozen sections of human muscle were separated by two-dimensional gel electrophoresis and detected by fluorography or Coomassie Blue staining. The major proteins were identical in different normal muscles obtained from either sex at different ages, and in Duchenne and myotonic dystrophy samples. Congenital myopathy denervation atrophy, polymyositis, and Becker's muscular dystrophy samples, however, showed abnormal myosin light chain compositions, some with a decrease of fast-fiber myosin light chains and others with a decrease of slow-fiber light chains. These protein alterations did not correlate with any specific disease, and may be cause by generalized muscle-fiber damage.

  10. Capillary electrophoresis-electrochemical detection microchip device and supporting circuits

    DOEpatents

    Jackson, Douglas J. (New Albany, IN); Roussel, Jr., Thomas J. (Louisville, KY); Crain, Mark M. (Georgetown, IN); Baldwin, Richard P. (Louisville, KY); Keynton, Robert S. (Louisville, KY); Naber, John F. (Prospect, KY); Walsh, Kevin M. (Louisville, KY); Edelen, John. G. (Versailles, KY)

    2008-03-18

    The present invention is a capillary electrophoresis device, comprising a substrate; a first channel in the substrate, and having a buffer arm and a detection arm; a second channel in the substrate intersecting the first channel, and having a sample arm and a waste arm; a buffer reservoir in fluid communication with the buffer arm; a waste reservoir in fluid communication with the waste arm; a sample reservoir in fluid communication with the sample arm; and a detection reservoir in fluid communication with the detection arm. The detection arm and the buffer arm are of substantially equal length.

  11. Electronic imaging systems for quantitative electrophoresis of DNA

    SciTech Connect

    Sutherland, J.C.

    1989-01-01

    Gel electrophoresis is one of the most powerful and widely used methods for the separation of DNA. During the last decade, instruments have been developed that accurately quantitate in digital form the distribution of materials in a gel or on a blot prepared from a gel. In this paper, I review the various physical properties that can be used to quantitate the distribution of DNA on gels or blots and the instrumentation that has been developed to perform these tasks. The emphasis here is on DNA, but much of what is said also applies to RNA, proteins and other molecules. 36 refs.

  12. Density gradient electrophoresis of cultured human embryonic kidney cells

    NASA Technical Reports Server (NTRS)

    Plank, L. D.; Kunze, M. E.; Giranda, V.; Todd, P. W.

    1985-01-01

    Ground based confirmation of the electrophoretic heterogeneity of human embryonic kidney cell cultures, the general characterization of their electrophoretic migration, and observations on the general properties of cultures derived from electrophoretic subpopulations were studied. Cell migration in a density gradient electrophoresis column and cell electrophoretic mobility was determined. The mobility and heterogeneity of cultured human embryonic kidney cells with those of fixed rat erythrocytes as model test particle was compared. Electrophoretically separated cell subpopulations with respect to size, viability, and culture characteristics were examined.

  13. Capillaries for use in a multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Chang, H.T.; Fung, E.N.

    1997-12-09

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  14. Monitoring environmental pollutants by microchip capillary electrophoresis with electrochemical detection

    SciTech Connect

    Chen, Gang; Lin, Yuehe; Wang, Joseph

    2006-01-15

    This is a review article. During the past decade, significant progress in the development of miniaturized microfluidic systems has Occurred due to the numerous advantages of microchip analysis. This review focuses on recent advances and the key strategies in microchip capillary electrophoresis (CE) with electrochemical detection (ECD) for separating and detecting a variety of environmental pollutants. The subjects covered include the fabrication of microfluidic chips, ECD, typical applications of microchip CE with ECD in environmental analysis, and future prospects. It is expected that microchip CE-ECD will become a powerful tool in the environmental field and will lead to the creation of truly portable devices.

  15. Capillaries for use in a multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S. (Ames, IA); Chang, Huan-Tsang (Silver Spring, MD); Fung, Eliza N. (Ames, IA)

    1997-12-09

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  16. Determination of benzylpenicillin in pharmaceuticals by capillary zone electrophoresis

    SciTech Connect

    Hoyt, A.M. Jr. ); Sepaniak, M.J. )

    1989-04-01

    A rapid and direct method is described for the determination of benzylpenicillin (penicillin G) in pharmaceutical preparations. The method involves very little sample preparation and total analysis time for duplicate results is less 30 minutes per sample. The method takes advantage of the speed and separating power of capillary zone electrophoresis (CZE). Detection of penicillin is by absorption at 228 nm. An internal standard is employed to reduce sample injection error. The method was applied successfully to both tablets and injectable preparations. 14 refs., 5 figs., 3 tabs.

  17. Analysis of mutations using PCR and denaturing gradient gel electrophoresis

    SciTech Connect

    Cariello, N.F.; Swenberg, J.A. Duke Univ., Durham, NC ); DeBellis, A.; Skopek, T.R. )

    1991-01-01

    Denaturing gradient gel electrophoresis (DGGE) separates DNA molecules based on primary sequence. Under the appropriate conditions, all base pair (bp) substitutions, frameshifts, and deletions less than about 10 bp can be resolved from the wild type sequence using DGGE. Polymerase chain reaction (PCR) permits facile amplification of a given region of the genome. The authors have combined PCR and DGGE to: (1) localize mutations in the X-linked human androgen receptor gene; (2) analyze thousands of thioguanine-resistant mutants simultaneously; (3) examine the fidelity of several DNA polymerases used in PCR.

  18. Polyacrylamide Gel Electrophoresis for Purification of Large Amounts of RNA.

    PubMed

    Meyer, Mélanie; Masquida, Benoît

    2016-01-01

    Polyacrylamide gel electrophoresis (PAGE) constitutes a powerful technique for the efficient purification of RNA molecules dedicated to applications that require high purity levels. PAGE allows for the fractionation of RNA obtained from cell extracts, chemical or enzymatic synthesis, or modification experiments. Native or denaturing conditions can be chosen for analytical or preparative-scale separations and the nucleotide resolution can be tuned by changing the percentage and reticulation of the gel material. In this protocol, we focus on the preparation of milligram-scale amounts of ~200 nucleotides (nt) RNA molecules that were used in subsequent crystallization experiments. PMID:26227037

  19. The fluid mechanics of continuous flow electrophoresis in perspective

    NASA Technical Reports Server (NTRS)

    Saville, D. A.

    1980-01-01

    Buoyancy alters the flow in continuous flow electrophoresis chambers through the mechanism of hydrodynamic instability and, when the instability is supressed by careful cooling of the chamber boundaries, by restructuring the axial flow. The expanded roles of buoyancy follow upon adapting the size of the chamber and the electric field so as to fractionate certain sorts of cell populations. Scale-up problems, hydrodynamic stability and the altered flow fields are discussed to show how phenomena overlooked in the design and operations of narrow-gap devices take on an overwhelming importance in wide-gap chambers

  20. Capillary electrophoresis: Biotechnology for separation of DNA and chromosomes

    NASA Technical Reports Server (NTRS)

    Williams, George O., Jr.

    1994-01-01

    Electrophoresis has been used for the separation of particles, ions, and molecules for a number of years. The technology for separation and detection of the results has many applications in the life sciences. One of the major goals of the scientific community is to separate DNA molecules and intact chromosomes based upon their different lengths or number of base pairs. This may be achieved by using some of the commercially available and widely used methods, but these processes require a considerable amount of time. The challenge is to achieve separation of intact chromosomes in a short time, preferably in a matter of minutes.